WL5 Abstract Book 3

Porto de Galinhas, Pernambuco, Brazil, May 13 th to 17th , 2013 Enotel

Abstract Book

Porto de Galinhas, Pernambuco, Brazil, May 13th to 17th, 2013 Enotel www.worldleish5.org

Abstract Book

Brazil – Pernambuco – Ipojuca

Fifth World Congress on Leishmaniasis

WorldLeish5 13th to 17th May 2013 Porto de Galinhas, Pernambuco, Brazil The World Congress on Leishmaniasis is the most important meeting on a disease of high priority in the World Health Organization’s list of Tropical Neglected Diseases. The leishmaniases are going to be around for a very, very long time because almost all of them have an animal reservoir, which excludes the possibility of elimination. It is therefore essential to define guidelines targeting the control. WL5 gathers leishmaniacs from all around the world.

Unlike other meetings it does not focus on one academic or public health field but on one group of diseases – the leishmaniasis.

Research on Leishmania parasites and the diseases they cause has increased over the last decade. Indeed, they have proven to be exciting models for studying parasite-host and vector-host interactions, immunological mechanisms, molecular biology, genetics, epidemiology and other related areas.

The principal aims of WorldLeish are to foster interaction between leishmaniacs from around the world and to provide a unique forum for discussing the state-of-the-art of leishmaniasis research, including the perspectives related to the control and prevention.

Besides this it allows participants to share their most recent findings and to meet and interact with some of the world’s leading experts.

The first World Congress on Leishmaniasis (WL1) was held in Istanbul, Turkey, in 1997 and was a great success. WL2 was held in Crete, Greece, in 2001, WL3 took place in Sicily, Italy in 2005 and WL4 took place in 2009 in Lucknow, India. The meeting’s success and popularity continued to increase as did the number of participants. However, they all took place in the Old World which made it difficult for younger American leishmaniacs to attend.

Now, after three meetings in Europe and one in Asia WorldLeish is finally taking place in an American country. Brazil represents one of most important leishmaniasis endemic areas, with the occurrence of many varieties of parasites which cause different clinical forms, including cutaneous, mucocutaneous and visceral leishmaniasis. Brazil has one of the world’s biggest critical masses of scientists and public health authorities working on leishmaniasis. This is due to the ever increasing importance of the disease and the levels of funding from different federal and state governmental bodies.

WL5 has broken all previous records. A total of 1,426 abstracts have been received which exceeded by far the number that was expected. Although a pleasant surprise, this created certain logistical problems. The challenge was how to accommodate so many high quality contributions as oral presentations since the number of sessions was limited. So, some had to be changed to posters. To help in analyzing these abstracts the Organizing Committee counted on both the International Scientific Committee and the Brazilian Scientific Committee, composed of 52 Brazilian leishmaniacs.

The final timetable has 5 simultaneous daily sessions with 1 satellite symposium, which to date is the most extensive WL program ever. Our actual Program introduces, besides 43 oral sessions and 14 Satellite Symposia, 22 special sessions designated as Hot Spots. The subjects for the Hot Spots are those that need greater focus or are considered to be controversial according to a survey performed on Leish-L. Abstracts in these specific areas were selected for presentation in the relevant Hot Spot. A total of 962 posters are presented in 3 evening sessions and specially selected posters will be located in 6 different areas, that are being called “Thinking Boxes”, to serve as discussion canters.

The number of participants has broken all records. Approximately 1,200 individuals from all walks of science and public health will be attending. We are grateful to the enthusiasm of all who have guaranteed WL5´s success, especially the younger ones from Latin America for whom this is possibly a once in a lifetime opportunity to attend a WL meeting.

We want to acknowledge our deepest gratitude to the organizations that have generously supported WL5 financially: Health Surveillance Secretary/Ministry of Health/Brazil, Oswaldo Cruz Foundation, Bill & Melinda Gates Foundation, Belgian Development Cooperation, NIH, PAHO/WHO, CNPq, CAPES, DECIT, FACEPE, and the Sponsorship of BAYER, SANOFI, ZOETIS/PFIZER, HERTAPE CALIER, PALADIN, BIOMANGUINHOS and MSD. Without their funding WL5 would probably not have taken place.

Due to committee’s dedication together with the generosity of funding agencies WL5 has been able to award registration waivers that include help with accommodation to 37 applicants selected from 15 countries were funds are scarce and where the disease is a significant public health issue.

We have put together a scientific program that has something for everybody and hopefully will open new horizons for both young and old leishmaniacs. We also wish that all of you enjoy the tranquility of Porto de Galinhas’s warm and clear waters, together with its excellent variety of local and international cuisine. These settings will create the perfect atmosphere for making new friends and reuniting with old ones. Have a wonderful time!

The Organizing Committee

- RESERVOIRS [361] P 697 - THE TARIM HARE (LEPUS YARKANDENSIS) AS RESERVOIR HOST OF LEISHMANIA INFANTUM IN DESERT-TYPE KALA-AZAR OF TARIM BASIN OF XINJIANG, CHINA LIAO, L.1; BATE, W.2; LUN, Z.3; YAN, S.1; XU, B.2; HOU, Y.2 1.CENTER FOR LABORATORY ANIMAL RESEARCH IN XINJIANG, URUMQI, CHINA; 2.CENTER FOR DISEASE CONTROL AND PREVENTION IN XINJIANG, URUMQI, CHINA; 3.SUN YAT-SEN UNIVERSITY, GUANGZHOU, CHINA.

Keyword:tarim hare (lepus yarkandensis); desert-type kala-azar; reservoir host

Abstract: Endemic foci and reservoir hosts of desert-type Kala-azar viceral leishmaniasis (VL) in Tarim Basin were investigated. Primary species idetification and distribution of animals in Tarim Basin were surveyed from 2004 to 2007, among which the predominant species were studied for their potential as reservoir hosts of desert-type viceral leishmaniasis (VL). These predominant animals were subjected to rk39-ELISA and rk39 dipstick to detect specific anti-leishmanial antibodies. Leishmania isolates from antibody-positive animals or from sandflies via microscopic examination were recovered by aseptic culture and expansion in animals, simultaneously. Natural infection rate of Leishmania in Phlebotomus wui (P. wui) and the predominant animal hosts for P. wui were also studied. Shrub habitats consist of diversifolious poplar or tamarisk were dorminated by tarim hare (Lepus yarkandensis), meridian gerbil (Meriones meridianas), Dipus sagitta and three-toed jerboa (Salpingotus crassicauda). 1284 samples from 16 animal species were analysed, among which only tarim hares (47/485) and domestic dogs (23/28) were positive for anti-leishmanial antibody. The natural infection rate of P. wui was 1.38%(10/726) and P.wui exclusively took blood meals from L. yarkandensis among 5 dominant wild animal species. Three Leishmania isolates were accquired from tarim hare (3/485) with another 6 ones from P. wui (6/726). During the study we found that the most suggested time for detection of naturally Leishmania-infected animals was from Autumn to Winter when the disease was developed in wild animals after sandfly season. The habitat mentioned above is suggested to be the infection source of desert-type VL for the following reasons: firsly, high density of sandflies with higher natural infection rate of Leishmania was found; secondly, 45% of L. yarkandensis was positive for anti-leishmanial antibody and Leishmania strains were also isolated in the Shrub habitats; thirdly, all new VL cases had a history of sandfly-biting in these habitats. For the first time, we had indentified L.yarkandensis as a reservoir host of L. infantum in desert-type VL in Tarim Basin in 2006. Evidences are based on the fact that P. wui exclusively take blood meal from L.yarkandensis, strains of Leishmania were isolated from L.yarkandensis and were identical to those from VL patient and the vector P. wui.

Correspondence to: Prof Li-fu Liao, Center for Laboratory Animal Research in Xinjiang, Urumqi 830002, P.R. China liaolif@21cn.com Acknowledgments Grants are supported by National Science Foundation of China #30460120

30760217

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- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [362] P 698 - EVALUATION OF SAND FLIES IN NOTIFICATION AREA WITH INDIGENOUS CASES OF CANINE VISCERAL LEISHMANIASIS, MARAMBAIA ISLAND, CITY OF MANGARATIBA, STATE OF RIO DE JANEIRO, BRAZIL. DE LIMA, D.A.; LOPES DO CARMO, L.A.; PONTES, C.S.; OSĂ“RIO, W.G.; ALVES, S.S.; DE SOUZA, M.B.; FIGUEIREDO, F.B.

FUNDAÇÃO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:american visceral leishmaniasis ; lutzomyia longipalpis; entomological survey

Abstract: Introduction:The leishmaniasis are zoonoses of major public health importance in Brazil and worldwide. These diseases are caused by parasites of the genus Leishmania that affect men and some species of animals. Transmission occurs via the bites of various kinds of phlebotomine vectors whose source of infection comprises different mammalian species. The main vector of American visceral leishmaniasis (AVL) in Brazil is the Lutzomyia longipalpis species, but other species have also been labeled as possible transmitters. In Marambaia Island, city of Mangaratiba, State of Rio de Janeiro are being reported numerous cases of canine visceral leishmaniasis, which causes great concern due to lack of knowledge on the epidemiological factors involving this outbreak. Therefore, this study aims to evaluate the sand fly fauna of this region and identify, through the technique of Polymerase Chain Reaction (PCR), species of sand flies infected with Leishmania (Leishmania) infantum chagasi, which are possibly involved in the transmission cycle of the disease. Materials and Methodos:The entomological survey began in August 2012. It is being conducted using suction light traps (CDC model), which are being introduced in peridomestic environment, such as chicken coops and kennels, and wild environment. Traps are being set at 6pm and taken off at 7am the next morning for a week, once a month, for a total of 65h of collections per trap. The specimens are placed in falcon tubes containing alcohol 70% properly identified, indicating the date and location of capture. Then the samples are being submitted to the Laboratory Institute of Rural Endemics - Fiocruz for screening and identification of species. Identified sampres are separated by species and sex to perform PCR in Clinical Research Laboratory in Dermatozoon in Livestock Research Institute Evandro Chagas - Fiocruz. Partial Results:The entomological survey conducted in this area has pointed sandfly Lutzomyia intermedia as the most frequent with, 280 males and 360 females captureds, followed by Lutzomyia migonei with 100 males and 30 females, while the main vector, Lutzomyia longipalpis is in low density until the moment of the survey with 4 males and 4 females. It is hoped that the results obtained in this study, to know the phlebotomine fauna of the region and possibly also the vector species involved in the transmission cycle of canine visceral leishmaniasis in the area.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [363] P 699 - AMERICAN CUTANEOUS LEISHMANIASIS: SURVEILLANCE OF VECTORS AND THE IMPACTS OF BRAZILIAN GOVERNMENT PROJECT (GROWTH ACCELERATION PROGRAM) IN COMMUNITIES OF RIO DE JANEIRO CITY, RJ, BRAZIL. COSTA, W.A.1; OSTRITZ, A.2; CORDEIRO, J.P.2; GOUVEIA, C.3; JORGE, T.A.1; RANGEL, E.F.1 1.IOC/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 2.CFMA/PRESIDENCIA/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.EXERCITO BRASILEIRO, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:sand-fly vectors; integrated control; american cutaneous leishmaniasis

Abstract: American Cutaneous Leishmaniasis (ACL) occurs by the association among links of epidemiological chain in restricted ecological niches, where ecological changes influence the rate of spreading of the disease through direct contact between man and the vector. In the Rio de Janeiro City, Brazil, the registration of cases of ACL has been increasing steadily since the 80's, mainly in the west of the city where the Campus Fiocruz Mata Atlântica – CFMA is located, in the district of Jacarepagua, where ACL affects communities with individuals of differing age

groups and sexes, with intra and peridomestic transmission. There is a consensus regarding the health-disease process on the relationship among the social life conditions and the development of diseases, requiring identifying indicators in biological and social spheres. The objectives of this study are: follow the process of the transmission of ACL, assessing impacts of the works of the Growth Acceleration Program on the dynamics of sand-fly vectors; implement integrated actions of entomological surveillance, environmental management practices, and health education; evaluate environmental indicators related to surveillance of vectors, especially the intra and peridomestic frequency ; map out the socio-economic profile of the residents and their access to health services, education, culture, and welfare and to correlate it with the occurrence of ACL. CFMA comprises an area of the Atlantic Forest, with a residential area equivalent to 45% coverage. The Monitoring Stations were established in communities of Caminho da Cachoeira, Fincão, and Sampaio Correa, where there is a high concentration of human cases of ACL and which will be most impacted by the initiatives of the government project. Systematic observations will be conducted in order to record changes in local features which may be related to the transmission cycle of ACL. Preliminary studies revealed that studied communities demonstrated the lowest social and economic indexes in the region, combining the environmental characteristics, which favor the dynamic differentiated transmission of the receptivity of each community and reflects on the distribution of human cases. The discussion of this data contributes to understanding the interface between health and environment, minimizing the risk of exposure of residents through the implementation of integrated actions of entomological surveillance, environmental management, and health education initiatives.

Financial support; Instituto Oswaldo Cruz-FIOCRUZ; CAPES - Brasil

- RESERVOIRS [365] P 700 - LEISHMANIA SPP. AND EHRLICHIA CANIS: CO-INFECTION OR CROSSREACTION IN BRAZILIAN DOGS? PAZ, G.F.1; AVELAR, D.M.1; KRAWCZAK, F.S.2; REIS, I.A.3; PINTO, M.C.1; PAGLIONI, D.N.4; PAIN, A.M.5; LABRUNA, M.B.2; WERNECK, G.L.6

1.CENTRO DE PESQUISAS RENÉ RACHOU/FIOCRUZ, BELO HORIZONTE, MG, BRAZIL; 2.UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 3.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 4.PREFEITURA MUNICIPAL DE BELO HORIZONTE, BELO HORIZONTE, MG, BRAZIL; 5.FUNDAÇÃO EZEQUIEL DIAS, BELO HORIZONTE, MG, BRAZIL; 6.UNIVERSIDADE FEDERAL DO RIO DE JANEIRO, RIO DE JANEIRO, MG, BRAZIL.

Keyword:dogs; leishmania; co-infection

Abstract: Leishmania spp. and Ehrlichia canis: co-infection or cross-reaction in Brazilian dogs?

Gustavo F. Paza, Daniel M. de Avelara, Felipe da Silva Krawczakb, Ilka A. Reis c, Marina C. C. Pinto a, Daniella N. Paglioni d , Andreza M. Pain e ,Marcelo B. Labruna b , Guilherme L. Werneck f

a

Centro de Pesquisas René Rachou/Fiocruz-MG, Av. Augusto de Lima 1715, 30190-002 Belo Horizonte, Brazil; b Universidade de São Paulo; c Universidade Federal de Minas Gerais/Instituto de Ciências Exatas; d Prefeitura Municipal de Belo Horizonte/Gerência de Controle de Zoonoses; e Fundação Ezequiel Dias; f Universidade Federal do Rio de Janeiro.

Ehrlichiosis and leishmaniasis are major vector-borne diseases of dogs with a worldwide distribution. Ehrlichia canis, a causative agent of acute or chronic monocytic ehrlichiosis is transmitted by the brown dog tick, Rhipicephalus sanguineus. Visceral leishmaniasis (VL) is an infectious disease that affects both human and animals. Domestic dogs (Canis familiaris) are principal reservoir hosts of VL in Brazil which is caused by Leishmania infantum (syn. L. chagasi). The epidemiology of canine ehrlichiosis and leishmaniasis overlaps in many areas of the world, because the vector activity and transmission periods of these pathogens are similar.

This study aimed to determine the frequency of infection and the possibility of co-infection or cross-reaction by both parasites in the Metropolitan Region of Belo Horizonte, Minas Gerais, Brazil, an endemic area of VL. Altogether, 96 serum samples were collected from domestic dogs from North West region of Belo Horizonte in August 2010. Serum samples were tested for the detection of anti-Leishmania IgG antibodies using three commercial canine visceral leishmaniasis kits (Bio-manguinhos速/Fiocruz/MS): indirect ELISA, indirect fluorescent antibody test (IFAT) and Dual Path Platform (DPP). Dogs were considered to be positive for Leishmania infection if found to be reactive by IFAT or DPP and ELISA (as recommended by the Brazilian Ministry of Health). For the detection of Ehrlichia canis infection, sera were tested with IFAT using E. canis antigen, strain S達o Paulo, with a cut-off titer of 1:80. The Fisher's exact test were used for statistical analyses (P<0.05). Of the 96 domestic dogs, 23 (23.9%) and 12 (12.5%) had antibodies against E. canis and Leishmania spp., respectively. Simultaneous infections of E. canis and Leishmania spp. was found in only one (1%) of the dogs. In VLpositive (12) and VL-negative dogs (84), E. canis was found in 8.3% and 26.2% respectively.

No significant correlation in E. canis seroprevalence was found when VL-positive or VLnegative dogs were compared (P>0.05). These findings suggest a pattern of haphazard coinfection in the endemic area and do not support the occurrence of serological cross-reaction among these parasites.

This study was financed by FAPEMIG /PPP and Fiocruz/PAPES VI - CELLULAR AND MOLECULAR BIOLOGY [366] P 701 - DNA LOAD ANALYSIS USING REAL TIME PCR IN COMPARISON WITH IMMUNOHISTOCHEMICAL FINDINGS OF DRY TYPE CUTANEOUS LEISHMANIASIS ;BEFORE AND AFTER TREATMENT BY IMIQUIMOD, GLUCANTIME AND COMBINATION OF BOTH DRUGS DABIRI, S.; MANAFI, H.; SHAMSIMEYMANDI, S.; FOTOUHIARDEKANI, R.; AMIRPOUR, S.; SHAMSIMEYMANDI, M. 1PATHOLOGY,2 DERMATOLOGY,3BIOINFORMATICS PASTEUR INSTITUETEHRAN,4PHARMACOLOGY DEPARTMENTS ,AFZALIPOU, KERMAN, IRAN, ISLAMIC REPUBLIC OF.

Keyword:leishmania tropica; real time pcr; glucantime, imiquimod

Abstract: Backgorund: Dry type cutaneous leishmanias (DTCL) is endemic in Bam a city of Kerman province, south east of Iran. This study was conducted for the first time in Kerman, using RealTime PCR method, in order to compare the effect of different treatments on parasite DNA load following therapy . Materials and Methods: Patients were divided into three groups under therapy with intralesional meglumine antimoniate , topical imiquimod and combination of both drugs. After obtaining consent from patients, punch biopsies were taken before and after treatment and paraffin blocks were provided .To compare the amount of DNA load a relative quantitative Real-Time PCR method was designed and set up using

L.tropica ITS(internal transcribed spacer) gene on paraffin blocks.To normalize a human gene (beta actin ) was used as internal control. After doing PCR ,the obtained results we analyzed using 2^- CT method and relativity of DNA load before and after treatment were calculated.

Results:The highest falling of DNA load was for Imiquimod (mean 4/7 cases), glucantime (mean 2/2 cases) and combination therapy (mean 2/4). From clinical point of view combination therapy had the best response. On the other hand the overall IHC findings showed good response based on decreased CD1a epidermal, increased CD1a dermal, decreased CD68 macrophages and increased CD3 and CD20 of dermis.

Conclusion: We concluded that imiquimod is immune modulator and has synergistic effects on the parasitocidal effect of glucantime for better response

- EPIDEMIOLOGY [368] P 702 - VISCERAL LEISHMANIASIS IN THE URBAN AREA OF NEIVA HUILA, COLOMBIA 2012 ZAMBRANO, C.D.P. INSTITUTO NACIONAL DE SALUD, BOGOTA, COLOMBIA.

Keyword:visceral leishmaniasis, ; urban focus,; leishmania infantum

Abstract: Introduction: Visceral Leishmaniasis is an endemic disease in Colombia with two historical transmission focus, mainly located in rural areas. The most important focus is found in the north of the country and the second in the departments of Cundinamarca, Tolima and Huila, the latest department, during the past 9 years has had an average of 1.5 cases per year, with a maximum of 4 cases during 2004 and a minimum of zero cases during 2008, 2009 and 2010.

During May of 2012, an outbreak of Visceral Leishmaniasis was confirmed in four communes of 3 districts of Neiva Huila, affecting seven (7) children.

Methodology: We conducted a study focus that included epidemiological research, surveys in order to identify risk factors, community and medical staff training, community and institutional active search of children under 5 years of age, entomological and domestic reservoirs studies (canines), wild reservoirs census and activities of promotion and control.

Results: The eco epidemiological conditions of Neiva allow the transmission of the disease.

The number of children in poverty, living in homes surrounded by primary and secondary forest and the nutritional deficiencies makes them more susceptible to the disease.

The unawareness of the disease, the lack of specific symptoms and the its course are factors that had a negative impact in the identification of the cases.

Diagnostic confirmation was made by direct examination in 85.7% of the cases and 14.7% by indirect immunofluorescence. The symptoms presented were fever and thrombocytopenia in 85.7% of the cases, anemia in 57.2% and hepatosplenomegaly in every case.

The 71.4% of the cases were girls and the 28.6% boys, aged between 10 months and 4 years.

100% of the cases were hospitalized and received treatment and there was not a lethal case. The

entomological study found Lutzomyia longipalpis in and around housing. The overall seroprevalence in canine reservoirs was 7% and the community addressed the presence of wildlife reservoirs.

There was residual spraying, distribution of bednets TILD, humanitarian elimination of canine and community training.

Conclusion: We identify a need for regular and sustainable programs for the surveillance and control of this disease, which involves early diagnosis and treatment, environmental management to control sandflies and reservoirs and a large component of community participation.

- EPIDEMIOLOGY [369] P 703 - ASSOCIATION BETWEEN INFESTATION BY ECTOPARASITES AND THE PRESENCE OF ANTI-LEISHMANIA ANTIBODIES: A COHORT STUDY OF DOGS FROM A BRAZILIAN ENDEMIC AREA PAZ, G.F.1; REIS, I.A.2; GOMES, V.A.3; AVELAR, D.M.1; FERREIRA, E.M.2; PINTO, M.C.1; ESTEVAM, L.M.1; WERNECK, G.L.4 1.CENTRO DE PESQUISAS RENÉ RACHOU/FIOCRUZ, BELO HORIZONTE, MG, BRAZIL; 2.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 3.PREFEITURA MUNICIPAL DE BELO HORIZONTE, BELO HORIZONTE, MG, BRAZIL; 4.UNIVERSIDADE FEDERAL DO RIO DE JANEIRO, RIO DE JANEIRO, MG, BRAZIL.

Keyword:dogs; flea; leishmania

Abstract: Association between infestation by ectoparasites and the presence of anti-Leishmania antibodies: A cohort study of dogs from a Brazilian endemic area Gustavo F. Pazª, Ilka A. Reis b, Varlei A. A. Gomes c, Daniel M. Avelar a, Elisa C. da Mata Ferreira b, Marina Caroline C. Pinto a, Letícia G. T. de Miranda Estevam a , Guilherme L. Werneck d a

Centro de Pesquisas René Rachou/Fiocruz - MG, Av. Augusto de Lima 1715, 30190-002, Belo Horizonte, Brazil; b Universidade Federal de Minas Gerais, Instituto de Ciências Exatas; c Prefeitura Municipal de Belo Horizonte, Gerência de Controle de Zoonoses; dUniversidade Federal do Rio de Janeiro It has been proposed that transmission of canine visceral leishmaniasis might involve the participation of mechanical vectors including ticks of the family Ixodidae, in particular Rhipicephalus sanguineus, and the cat flea Ctenocephalides felis felis. The association between the infestation by R. sanguineus and C. felis felis and the presence of anti-Leishmania antibodies has been evaluated in a cohort study of dogs living in a Brazilian endemic area for canine visceral leishmaniasis. Blood samples from 88 seronegative domestic dogs were submitted to indirect immunofluorescent antibody assay every three months, for one year. Dogs that turned seropositive (n = 20) as well as those that remained seronegative (n = 68) were evaluated monthly for the presence of ticks and fleas, the results of which were expressed qualitatively as infested or non-infested, irrespective of the intensity of infestation. At the end of follow-up, all animals were asymptomatic and all seropositive animals were sacrificed. The odds of infection was significantly greater (OR=3.54,CI95%, 1.10 - 12.53) among dogs infested by C. felis felis as compared with their non infested counterparts. Unlike, the odds of infection showed no significance difference between groups infested and non-infested by R. sanguineus (OR=0.31;

CI95%, 0.03 - 1.52). The study provides further evidence for the potential role of C. felis felis in mechanically transmitting Leishmania to the canine population.

This study was financed by Fundação de Amparo à Pesquisa de Minas Gerias (FAPEMIG – Programa PPP) and Programa de Apoio à Pesquisa da Fundação Oswaldo Cruz (PAPES VI).

- CONTROL PROGRAMS [371] P 704 - KNOWLEDGE ASSESSMENT ABOUT CANINE AND FELINE LEISHMANIASIS OF PET´S OWNERS IN ASUNCIÓN, PARAGUAY GROPPI, S.1; ACUÑA, V.L.1; ALARCÓN, G.1; MIRET, J.2

1.FACULTAD DE CIENCIAS VETERINARIAS . UNIVERSIDAD NACIONAL DE ASUNCIÓN (UNA), SAN LORENZO, PARAGUAY; 2.INSTITUTO DE INVESTIGACIONES EN CIENCIAS DE LA SALUD (IICS). UNIVERSIDAD NACIONAL DE ASUNCIÓN (UNA), ASUNCIÓN, PARAGUAY.

Keyword:canine; feline ; leishmaniasis

Abstract: Dogs and cats provide companionship to theirs owners helping with their psychological health; however, despite of the important roles played by pets in human wellbeing, they are an important sources of zoonotic infections. Canine visceral leishmaniasis is an important endemic disease in Paraguay. The objective of this observational, descriptive and prospective study was to determine the knowledge of 160 pet´s owners about canine and feline leishmaniasis. A questionnaire was applied to obtain information about vector transmission to humans and animals and prevention methods in pets, how pet´s owner had the knowledge and if some family member suffered this disease. It was observed that 153 pet´s owners (95.6%) know the disease, and only 4 pet´s owners (4.3%) did not hear about leishmaniasis. 154 pet´s owners (96.2%) affirm that sandflies bite are responsible for the transmission to humans and pets. (88.6%) of the pet´s owners know that dogs and cats could transmit the disease to human. About the prevention, 29 pet´s owners (18.1%) remove organic waste and the breeding ground of sandfly, (10,6%) use wire mesh and fumigate the house and yard, (10,6%) use collar and spot-on insecticide on their pets, (30.6%) apply the three methods aforementioned, (20%) apply the culling of positive dog, (6.8%) use canine leishmaniasis vaccines. (39.3%) pet´s owners had knowledge about leishmaniasis by newspaper, television and radio, (28.1%) in the school and (21.2%) were inform by the veterinarians. Nobody of the pet´s owner family members suffered leishmaniasis yet. In spite of the interviewee number it is not a representative universe of the pet´s owners that goes to the veterinary clinic in Asunción, Paraguay; this study gives an idea of the knowledge about this disease and the necessity to improve the communication between the pet´s owners, veterinarian clinician and public health organization about the knowledge and prevention of this zoonotic disease.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [373] P 705 - A PRELIMINARY MODEL OF CLIMATIC SUITABILITY FOR LUTZOMYIA FLAVISCUTELLATA (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE), MAIN VECTOR OF LEISHMANIA AMAZONENSIS IN BRAZIL, UNDER CLIMATE CHANGE CARVALHO, B.M.1; VALE, M.M.2; RANGEL, E.F.3 1.UNIVERSIDADE DO ESTADO DO RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 2.UNIVERSIDADE FEDERAL DO RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 3.INSTITUTO OSWALDO CRUZ/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:lutzomyia flaviscutellata; distribution modeling; climate change

Abstract: Global climate change is likely to affect vector borne diseases, through environmental changes that could lead to distribution shifts. In this dynamic process, new areas suitable for transmission may appear. Lutzomyia flaviscutellata is responsible for the transmission of New World Diffuse Cutaneous Leishmaniasis (DCL), a severe form of the disease caused by Leishmania amazonensis. The disease occurs in north and central South America, with a especially high incidence in the Amazon. In this preliminary study, climatic suitability was modeled for L. flaviscutellata under a future climate change scenario using the Maximum Entropy approach.

Were used 81 occurrence records for L. flaviscutellata in Brazil, gathered from the literature and state health departments, together with 15 bioclimatic variables under a “business as usual” (A2a) greenhouse gas emission scenario for 2050. The resulting model had a good performance (AUC = 0.93), but was strongly biased towards highly sampled areas along cities, roads and rivers. It showed a contraction of climatically suitable areas in Amazon (north), coupled with a slight expansion of those areas in the Atlantic forest (south). This southeastern expansion is of public health concern. Autochthonous human cases of DCL in southeast Brazil are currently rare, although since 1980's the disease seems to be gradually expanding its occurrence southward. These results should be interpreted with caution, however, because the model was built with data from Brazil only. Next steps include adding occurrence records from other countries and other environmental variables such as land use and land cover change, and run these models on different algorithms, using ensemble modeling to obtain better predictions of the species current and future distributions.

- EPIDEMIOLOGY [374] P 706 - NEW FOCUS OF VISCERAL LEISHMANIASIS IN URBAN AREA OF CARTAGENA,COLOMBIA 2011 ZAMBRANO, C.D.P. INSTITUTO NACIONAL DE SALUD, BOGOTA, COLOMBIA.

Keyword:visceral leishmaniasis; study focus; leishmania infantum

Abstract: Introduction: Visceral leishmaniasis is a endemic disease in Colombia with two historical and characterized transmission focus, the largest located in the departments of Bolívar, Sucre and Córdoba, and the second in the departments of Cundinamarca, Tolima and Huila. It is a disease with low morbidity and a significant impact on mortality. During the January of 2011 a case of visceral leishmaniasis was confirmed in a one year old girl from the neighborhood Lo Amador in Cartagena. The child never visited risk areas.

Methodology: We conducted a Study Focus that included epidemiological research, 45 surveys were applied to identify risk factors associated with housing characteristics, knowledge about the disease and activities of prevention, community and institutional active search of children under 15 year of age for parasitological diagnosis through indirect immunofluorescence (IIF), entomological study and study of domestic (dogs) and wild reservoirs of the disease.

Results: The proximity of housing to the wildlife focus in El Cerro de la Popa, the precarious sanitation conditions, the lack of symptoms and the mode of transmission of the disease are major risk factors. The case presented prolonged fever, anemia, thrombocytopenia and hepatosplenomegaly, it was confirmed by indirect immunofluorescence IFI 1:32. The girl was hospitalized and received treatment with clinical improvement. In the communitarian active search were identified three children with IFI titles below 1:32 (not reactive), two of them with evidence of Reactive Montenegro, and four adults with Reactive Montenegro test, which is evidence of previous contact with the parasite‘s antigens. The study found Lutzomyia evansi

entomological vector confirming the disease in and around housing. The canine reservoirs overall seroprevalence was 30.7% and a marsupialis Didelphys confirmed by PCR and histopathology compatible for Leishmania.

Conclusion: The characterization of a new focus of Visceral Leishmaniasis in the country calls for the intensification and strengthening of disease surveillance in the whole country, emphasizing in early detection, prompt treatment and follow-up of cases in order to reduce underreporting and mortality for this cause.

- CONTROL PROGRAMS [376] P 707 - SEROPREVALENCE AND EPIDEMIOLOGY OF CANINE VISCERAL LEISHMANIASIS IN PARAGUAY MIRET, J.; SOSA, L.; GALEANO, E.; OCAMPOS, H.; MARTÍNEZ, R.; OJEDA, J.; CASTAGNINO, M. NATIONAL PROGRAMME OF ZOONOSES CONTROL AND NATIONAL RABIES CENTER (PNCZYCAN), SAN LORENZO, PARAGUAY.

Keyword:canine; leishmaniasis; paraguay

Abstract: Dogs are considered to be the main domestic reservoirs of the parasite Leishmania infantum, they play an important role in the epidemiological cycle of visceral leishmaniasis transmission to human hosts. Canine leishmaniasis is an endemic disease and highly prevalent in Paraguay.

The purpose of this work was to determine the seroprevalence of canine leishmaniasis by immunochromatographic rK39 test in serum samples obtained for routine exam requested by dog´s owners and veterinarians, active surveillance in areas of silence transmissions and control of human cases notified for the National Service of Malaria Erradication (SENEPA) to the National Programme of Zoonoses Control and National Rabies Center (PNCZyCAN) of the Ministry of Public Health and Social Welfare (MSPyBS) in 2012. A total of 12517 canine blood samples were analyzed by immunochromatographic rK39 test (Cypress Diagnostics®, Belgium), in the laboratory of Leishmaniasis of the PNCZyCAN coming from 12/17 departments of the Paraguay. The statistical results were analyzed by Epiinfo 3.5.1. The 5340 blood samples coming from routine exam from Asunción (capital city) and the departments: Central, Boquerón, Caaguazú, Cordillera, Itapúa, Guairá, Presidente Hayes, Paraguarí, Misiones, Alto Paraná and Caazapá showed 2405 positive serum samples with a prevalence of (45.03%) of canine leishmaniasis. The active surveillance coming from Asunción and the departments: Central and Misiones showed that 350 out of 2270 dog´s blood samples were identified as positive with a prevalence of (15.41%) of canine leishmaniasis. From the 56 focus of human visceral Leishmaniasis cases, coming from Asunción, and the departments Central, Cordillera and Paraguarí, it was observed that 791 out of 4907 dog´s blood samples showed a prevalence of (16.11%) of canine leishmaniasis. It was observed a global prevalence of (28.32%). Euthanasia procedures were performed in 273 positive dogs (78%) coming from active surveillance in areas of silence transmission and 492 positive dogs (62.19%) coming from focus of human visceral leishmaniasis. The high canine visceral leishmaniasis shows the urgent need to continue a strict epidemiological surveillance, sanitary education and community participation by the Ministry of Public Health and Social Welfare (MSPyBS) in the control of this disease in Paraguay.

- EPIDEMIOLOGY [379] P 708 - PROGNOSTIC FACTORS FOR DEATH FROM VISCERAL LEISHMANIASIS IN BRAZIL: A SYSTEMATIC REVIEW AND META-ANALYSIS BELO, V.S.1; WERNECK, G.L.1; BARBOSA, D.S.1; HORTA, M.A.P.1; DA SILVA, E.S.2; NASCIMENTO, B.W.L.3; STRUCHINER, C.J.1

1.ESCOLA NACIONAL DE SAÚDE PÚBLICA - FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 2.UNIVERSIDADE FEDERAL DE SÃO JOÃO DEL REI, DIVINÓPOLIS, MG, BRAZIL; 3.CENTRO DE PESQUISAS RENÉ RACHOU, FIOCRUZ, BELO HORIZONTE, MG, BRAZIL.

Keyword:visceral leishmaniasis; prognostic factors; death

Abstract: An important way to reduce fatality rates of visceral leishmaniasis is to early identify markers of bad prognosis or death. The objectives of the present study were to evaluate the role of such potential prognostic factors that have been investigated in the Americas and to determine the gaps in existing knowledge. Literature searches were carried out using four databases (Pubmed/Medline, Lilacs, CAPES Databank and Google Scholar), the reference lists within articles, and references provided by experts. A tool for the evaluation of limitations and susceptibilities to bias of publications was developed. The information of each of the studied prognostic factors was combined by meta-analyses of p-values or of effect sizes. The I2 statistic was used to assess heterogeneities among the studies. Prognostic factors were organized by level of evidence taking into account: the summary measures obtained in meta-analysis, the patterns of associations and the results of multivariate analyses performed in primary studies. Fourteen publications carried out in Brazil were selected for detailed review. In eleven, the main sources of data collection were medical charts. In two studies, patients were followed up after hospitalization and in one study the data source was the Information system of notifiable diseases of the Brazilian Ministry of Health. Except for the time of onset of symptoms to hospitalization and gender, the prognostic factors operated in the same way and in the same direction regardless of age group. Prognostic factor showing of strong evidence were: jaundice (Odds Ratio (OR): 6.3; 95% confidence interval (CI): 3.9 - 10.3), low platelet count (OR:5.5; CI:3.0 – 10.3), age younger than 5 (OR:2.7; CI:1.6 – 4.7) or older than 40 years (OR:3.1; CI:2.3 – 4.3), severe neutropenia (OR:2.3; CI:1.5 – 3.6), presence of bleeding (OR:5.0; CI:3.5 – 7.2), coinfection with HIV (OR:3.7; CI:2.1 – 3.6), diarrhea (OR:2.8; CI:2.0 – 3.9), dyspnea (OR:3.2; CI:2.3 – 4.6) and infection (OR:3.1; CI:2.4 – 4.1). Several other variables were analyzed in few studies and were not included in the classification of evidence adopted. The main limitation of existing knowledge was the lack of validation of the few prediction models developed. The results described might be used as an aid to clinical decision-making in the appropriate identification and targeting of patients at higher risk. Future studies should produce more consistent data and prognostic models externally validated.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [380] P 709 - COMPOSITION OF SANDFLY SPECIES IN A VISCERAL LEISHMANIASIS FOCUS IN AN URBAN AREA OF CARTAGENA DE INDIAS, COLOMBIA FUYA, P.1; AYALA, M.1; ZAMBRANO, P.1; CANTILLO, O.2 1.INSTITUTO NACIONAL DE SALUD, BOGOTÁ, COLOMBIA; 2.UNIVERSIDAD DE ANTIOQUIA, MEDELLIN, COLOMBIA.

Keyword:visceral leishmaniasis; urban area; cartagena, colombia

Abstract: Introduction In Colombia leishmaniasis is endemic and annually are reported averaged 14,000 cases. In the Colombian Caribbean region, not only the cutaneous but the visceral form are a major public

health problem, cases are reported mainly in rural areas, although recent years have been presented more and more often cases in urban areas. In January 2011 we confirmed the presence of one case of in a girl of one year in an urban district Lo Amador in the Cartagena city (Bolívar). The purpose of this paper is to present the results of entomological study conducted with the aim of confirming the species composition of sandflies presents in this area.

Methodology Sandflies were collected in March 2011, in ten houses, including the patient's home (10 ° 25'194 "N and 75 ° 31'768" W), at a height above sea level between 14 and 69 meters. In each house installed two CDC miniature light traps placed in the intra and peridomestic habitats, for three consecutive nights. Each household chose the room where slept the greater number of people, around the homes were selected courtyards and outhouses where pets slept. Traps were activated from 18:00 and retired at 6:00. In the morning, between 6:00 and 7:00 sandflies search was made to stand in the inner and outer walls of the houses. Sandflies collected were preserved in 70% alcohol, for taxonomic determination rinsed with potassium hydroxide and 10% liquid phenol and used the key to Young and Duncan.

Results Were collected in total 113 sandflies, the most abundant species was Lutzomyia (Micropygomyia) trinidadensis with 89 specimens, followed by Lutzomyia (verrucarum) evansi with 13 specimens, this specie is a vector of Leishmania chagasi, causative agent of visceral leishmaniasis, Lutzomyia (Micropygomyia) cayennensis cayennensis with 7 specimens and Lutzomyia (Micropygomyia) venezuelensis with 4 specimens. Lutzomyia (verrucarum) evansi was collected in the bedroom in one of the houses and their peridomestic areas in three homes, Lutzomyia (Trichopygomyia) trinidadensis and Lutzomyia (Micropygomyia) cayennensis cayennensis were collected both within and around the homes and Lutzomyia (Micropygomyia) venezuelensis only in peridomestic area, as shown in the following table.

Table 1. Distribution of sandflies in athe intra and peridomestic areas Conclusion In the urban focus of visceral leishmaniasis in the city of Cartagena, was reported the presence of four species of sandflies, including Lutzomyia (verrucarum) evansi, confirmed vector of visceral leishmaniasis, Micropoygomyia cayennensis cayennensis, which has been found with unidentified flagellates, Micropygomyia trinidadensis and Trichopygomyia venezuelensis, the latter two species have no known medical importance. The finding of Trichopygomyia venezuelensis is the first report of this specie for the department of Bolivar, Colombia. These species, except Trichopygomyia venezuelensis, hav - RESERVOIRS [382] P 710 - WILD HOST INVOLVED IN A VISCERAL LEISHMANIASIS EPIDEMIC FOCUS IN CARTAGENA DE INDIAS, COLOMBIA.2011 ZAMBRANO, C.D.P. INSTITUTO NACIONAL DE SALUD, BOGOTA, COLOMBIA.

Keyword:reservoirs sylvatic; visceral leishmaniasis; didelphis marsupialis

Abstract: Leishmaniasis outbreak studies, require the characterization of key elements involved in transmission: Infected vectors and hosts. However, one of main problems for understanding the

ecoepidemiology of this disease is the lack of knowledge of natural reservors of the parasite which is key at the time of studying an outbreak of cutaneous and visceral leishmaniasis. . The purpose of this work was to identify wild mammals infected with Leishmania sp in one focus that took place in Cartagena de Indias, Colombia, in February 2011. This study was carried shortly after the outbreak in March 2011 in the Lomador neighborhood situated at the southwestern side of La Popa hill in Cartagena de Indias. The study design consisted of 4 Transects with 15 Tomahawk® and 18 Sherman live trap baited with universal bait that were set over a 3 night period. Which implied an effort of 99 traps/night. Captured wild animal were anesthetized and sacrificed for sample collection of liver, spleen and lymphatic ganglion. From each organ a sample tissue was evaluated by PCRtargeting a 120bp fragment of the kDNA conserved area, One adult male Didelphis marsupialis, was captured in the highest hills zone at 200 Mts of the index-case-house. This marsupial showed infection by Leishmania sp. in all organs evaluated. In this work we corroborate the presence of an infected wild host near a domestic focus of visceral leishmania D. marsupialis presents synanthropic and nomadic behavior facilitating the occurrence of outbreaks and establish a connection between sylvatic and domestic transmission cycles. This species has been previously reported like as Leishmania ssp and T. cruzi. reservoir and thus it could be involved in the outbreak focus in Cartagena de Indias. However, the role of this species should be further established increasing the sample size and characterizing the parasite strains.

- EPIDEMIOLOGY [384] P 711 - POPULATION STRUCTURE AND EVIDENCE OF PANMIXIS AMONG BRAZILIAN STRAINS OF THE LEISHMANIA (VIANNIA) SUBGENUS KUHLS, K.1; CUPOLILLO, E.2; SILVA, S.O.3; SCHWEYNOCH, C.4; CORTES BOITÉ, M.2; MELLO, M.N.3; MAURICIO, I.5; MILES, M.6; WIRTH, T.7; SCHOENIAN, G.4

1.MOLECULAR BIOTECHNOLOGY AND FUNCTIONAL GENOMICS, TECHNICAL UNIVERSITY OF APPLIED SCIENCES, WILDAU, GERMANY; 2.LABORATÓRIO DE PESQUISA EM LEISHMANIOSE, INSTITUTO OSWALDO CRUZ - FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 4.INSTITUTE OF MICROBIOLOGY AND HYGIENE, CHARITÉ UNIVERSITY MEDICINE BERLIN, BERLIN, GERMANY; 5.INSTITUTO DE HIGIENE E MEDICINA TROPICAL, LISBOA, PORTUGAL; 6.DEPARTMENT OF PATHOGEN MOLECULAR BIOLOGY, LONDON SCHOOL OF HYGIENE AND TROPICAL MEDICINE, LONDON, UNITED KINGDOM; 7.MUSÉUM NATIONAL D'HISTOIRE NATURELLE, DEPT. DE SYSTÉMATIQUE ET ÉVOLUTION, PARIS, FRANCE.

Keyword:microsatellite typing; population genetics; molecular epidemiology

Abstract: Parasites of the subgenus Leishmania (Viannia) are capable of causing a variety of clinical symptoms ranging from cutaneous leishmaniases with single or few lesions, including disseminated CL with multiple lesions to disfiguring forms of mucocutaneous leishmaniasis. In this population genetics study, a total of 120 Leishmania (Viannia) spp. strains from different endemic foci in Brazil were examined for variation at 15 hyper-variable microsatellite markers: 37 strains of L. guyanensis, 63 of L. braziliensis, four of L. shawi, six of L. lainsoni, seven of L. naiffi, one each of L. utingensis and L. lindenbergi, and one L. lainsoni/L. naiffi hybrid detected by isoenzyme analysis. The multilocus microsatellite profiles obtained were analysed using both model and distance based methods.

Significant genetic diversity was observed and almost all strains presented unique microsatellite profiles. Different types of population genetic analyses, including Bayesian inference, distance based and factorial correspondence analysis, as well as F statistics, revealed the existence of two well-defined populations in the sample set. Population 1 consisted of L. guyanensis strains from the Amazon region and Population 2 of strains of L. braziliensis isolated along the Atlantic coast

of Brazil. A third group was extremely polymorphic and included the other species of the subgenus Leishmania (Viannia) (L. shawi, L. naiffi, L. lainsoni, L. utingensis, and L. lindenbergi) from the north and north west of Brazil, as well as L. braziliensis strains also mainly from northern and western regions. This third group may represent an ancestral lineage and might have given rise to two new populations through bottleneck events (Populations 1 and 2). The Amazon forest seems to be the region of origin (and possibly speciation) with population migrations to the east and north. Surprisingly, the population of L. braziliensis from the Atlantic coast, and, to a lesser extent, that of L. guyanensis, were found to be close to panmixia. Random mating could be an explanation for the tremendous genetic diversity but limited population structure found in the L. (Viannia) subgenus in Brazil.

- EPIDEMIOLOGY [396] P 712 - SEROLOGICAL MONITORING OF NEW CASES OF CANINE VISCERAL LEISHMANIASIS IN MARAMBAIA ISLAND, CITY OF MANGARATIBA, RJ LOPES DO CARMO, L.A.; DE SOUZA, T.L.; DE ALMEIDA, A.B.; SANTOS, F.N.; FILHO, C.J.L.B.; PONTES, C.S.; SILVA, V.L.; DE SOUZA, M.B.; FIGUEIREDO, F.B. FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:monitoring ; canine visceral leishmaniasis; lutzomyia longipalpis

Abstract: Introduction: The American Visceral Leishmaniasis (AVL) in Brazil is caused by Leishmania (Leishmania)infantum chagasi, the main vector of the parasite is the Lutzomyia longipalpis, and the reservoir in the domiciliary area, dogs, both of which contribute to the maintenance of the disease cycle. For the diagnosis of leishmaniasis should consider the association between clinical, laboratory and epidemiological studies. Canine visceral leishmaniasis (CVL) presents clinically in various ways. Laboratory diagnostics include parasitological methods with direct microscopic identification and isolation of the parasite, and serological tests such as indirect immunofluorescence assay (IFA) and enzyme immunoassays, enzyme-linked immunosorbent assay (ELISA). For the diagnosis of CVL, the Ministry of Health recommends screening ELISA and IFA for confirmation of cases. This study assessed the prevalence of CVL in 2012 through canine serosurvey in Marambaia Island, city of Mangaratiba, State of Rio de Janeiro and the sand fly fauna of the region. Materials and methods: To evaluate serological tests were used in ELISA, IFA and immunoassay (DPP), a commercial kit produced by Bio-Manguinhos ÂŽ. For survey of sand flies were captured by CDC traps. Results: The census totaled 116 dogs, 17 dogs being identified positive, a prevalence of 14.65%. We found 9 species of sandflies, Lutzomyia longipalpis, Nyssomyia intermediate Migoneimyia migonei, Pintomyia fischeri, Evandromyia edwardsi, Micropygomyia capixaba, Pintomyia bianchigalatiae, Micropygomyia schreiberi and Bruptomyia sp., a total of 1.730 specimens captured in the period April-November 2012.

Conclusion: The confirmation of 17 (14.65%) seropositive dogs for LVC in Marambaia Island in 2012, reveals the maintenance of disease prevalence in 2009 was 15%, even with the removal of most dogs sororeatores diagnosed in 2009. In entomological survey found a low density of L. longipalpis in the region, which can not explain the finding of human cases. Failure to reduce the prevalence of LVC in the region, may be related to use of control measures in isolation.

Another important point that can be corroborated inefficiency to control is the environment of the region in which the houses are located in the woods, where the presence of vector and animal reservoirs is abundant.

- EPIDEMIOLOGY [407] P 713 - SEROPREVALENCE OF AMERICAN CUTANEOUS LEISHMANIASIS IN HUMANS AND DOGS IN A SLUM COMMUNITY OF JATAIZINHO, PARANĂ , BRAZIL

CALDART, E.T.1; BENITEZ, A.D.N.1; SOUZA, V.A.1; CONSTANTINO, C.1; ABATE, H.L.2; EVERS, F.1; GONÇALVES, D.D.3; DE FREITAS, J.C.1; FREIRE, R.L.1; NAVARRO, I.T.1

1.UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL; 2.UNIVERSIDADE FILADELFIA, LONDRINA, PR, BRAZIL; 3.UNIVERSIDADE PARANAENSE, UMUARAMA, PR, BRAZIL.

Keyword:seroprevalence; leishmaniasis; human

Abstract: Dogs and humans share the same environment and show similarities regarding infection by Leishmania spp, so dogs can be sentinels to evaluate the presence and spread of this parasite. In the Paraná State, Southern Brazil, autochthonous transmission of American cutaneous leishmaniasis (ACL) is found in many regions including Northern, where Jataizinho is located.

This work aimed to determine the prevalence of anti-L.amazonensis IgG antibodies in dogs and humans from a slum community of Jataizinho. In July 2010, 102 households from ten different blocks, characterized as invasion area, were visited and blood samples from dogs and their owners were taken. A questionnaire with variables about environment, sanitation and dog conditions was applied and serum samples were submitted to Indirect Fluoresce Antibody Test (IFAT). Titers 40 were considered positive for dogs and humans. Out of 167 dogs and 73 humans, 16 (9.6%) and four (5.5%) were positive to L. amazonensis, respectively. Data analysis revealed no association of canine leishmaniasis and variables researched. In the same direction, human and dog serological positivity was not statistically associated. However, half of positive human (two) are owners of positive dogs, this information reinforces the suspicion of the presence of the vector in these specific residences. The lack of statistical association is probably due to the low number of positives. Furthermore three of four positive humans live on the same block.

- OPERATIONAL HEALTH [410] P 714 - EVALUATION OF EXTRACELLULAR MATRIX IN THE EAR SKIN OF DOGS NATURALLY INFECTED WITH LEISHMANIA INFANTUM CARDOSO, J.M.O.1; VIEIRA, P.M.A.1; OLIVEIRA, F.C.B.1; DUZ, A.L.C.1; VITAL, W.C.2; GIUNCHETTI, R.C.2; REIS, A.B.1; CARNEIRO, C.M.1 1.UNIVERSIDADE FEDERAL DE OURO PRETO, OURO PRETO, MG, BRAZIL; 2.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL.

Keyword:leishmaniasis; matrix extracellular; dogs

Abstract: Canine visceral leishmaniasis (CVL) is a severe zoonotic disease of dogs caused by protozoa of the Leishmania donovani complex. Dogs are the domestic reservoir for human visceral leishmaniasis and many of the clinical and pathologic signs observed in dogs are similar to the pattern of the disease in humans. The skin is the first point of contact with Leishmania and was considered to be an important reservoir compartment for parasites in healthy and sick infected dogs and the important role of dogs in VL transmission is supported by the high parasite loads found in the skin of infected animals. Thus, the aim of this work was study the extracellular matrix alterations in the ear skin of dogs naturally infected with Leishmania (Leishmania) infantum. In order to do this, thirty six dogs naturally infected were categorized as asymptomatic (n=12), oligosymptomatic (n=12) and symptomatic (n=12). Eight non-infected dogs were used as control group. During necropsy, ear skin fragments were collected and fixed in formaldehyde solution. Paraffined sections of the tissues were stained with Picrosirius red, Masson’s Trichrome, Hematoxylin-Eosin (H&E) and analyzed by optical microscopy using the software Leica Qwin V3. All animals showed an inflammatory reaction ranging from mild to intense, and the cellular exudate consisting mainly of mononuclear cells. The inflammation was higher in symptomatic dogs when compared to the other groups. In the analysis of collagen area,

it was observed that the more intense the inflammation, like the presented by the symptomatic dogs, the lower area of collagen was noticed.In the Picrossirius red analysis, a reduction of collagen type 1 fibers was observed in symptomatic animals when compared to control group.

There was also an increase in type III collagen fibers in oligosymptomatic dogs compared to control dogs. Since this compartment is formed mainly by collagen fibers, it can be assumed that the presence of amastigotes may be involved in the degradation of the extracellular matrix.

These data suggest that there was injury in the skin of oligosymptomatic animals but they managed to repair this damage. However, the symptomatic animals were injured but they could not overcome the injury, perhaps because the higher inflammatory process observed in these animals. In this sense, the skin should be considered as a good place for parasitological analysis, being a reliable indicator for the severity of clinical disease in the CVL. "Supported by: CAPES, FAPEMIG, CNPq e UFOP" - EPIDEMIOLOGY [412] P 715 - SEROEPIDEMIOLOGICAL SURVEY ON AMERICAN CUTANEOUS LEISHMANIASIS IN DOGS FROM URBAN AREA OF JATAIZINHO MUNICIPALITY, PARANÁ, BRAZIL CALDART, E.T.1; BENITEZ, A.D.N.1; RODRIGUES, W.B.1; ABATE, H.L.2; GONÇALVES, D.D.3; DE FREITAS, J.C.1; FREIRE, R.L.1; NAVARRO, I.T.1 1.UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL; 2.UNIVERSIDADE FILADELFIA, LONDRINA, PR, BRAZIL; 3.UNIVERSIDADE PARANAENSE, UMUARAMA, PR, BRAZIL.

Keyword:seroprevalence; leishmaniasis; dog

Abstract: Infected dogs play an important role in public health as sentinels for American cutaneous leishmaniasis (ACL) agents on the environment allowing to evaluate the spread of this disease.

This work aimed to determine the prevalence of anti-L. amazonensis IgG antibodies and epidemiological variables associated to infection in dogs from Jataizinho, a small town in Paraná, Brazil. The town was divided in five urban regions used for strategic popular health programs and 357 residences, from 42 blocks selected randomly, were visited from July to November in 2010. Blood samples of 628 dogs were taken. A questionnaire was applied and serum samples were submitted to Indirect Fluorescence Antibody Test (IFAT), titers 40 were considered positive. The canine population was consisted of: 44.9% females; 78.9% undefined breed; 13.6% aged less than 1 year. Out of 628 animals, 46 (7.8%) were positive suggesting a consistent prevalence of anti-L. amazonensis IgG antibodies in this area. Analysis of variables showed significant difference with weight loss on the last 30 days. Weigh loss is not the major clinical sign found in dogs with ACL, but is not unusual. In urban region of Jataizinho we found 94.3% of the residences with basic sanitation, 93.4% with garbage collection and 100% with treated water. On the other hand, 41.3% accumulate rubble on the yard, 74.0% reported rodents in home, 39.1% are nearby a wasteland, 17.0% are next rivers and 13.6% are beside a forest.

These environmental characteristics are important because is known that they facilitate, directly or indirectly, the reproduction of the vector responsible for the transmission of ACL, but in this work they are not statistically associated with the risk of having the disease.

- EPIDEMIOLOGY [415] P 716 - AMERICAN CUTANEOUS LEISHMANIASIS SERODIAGNOSTIC IN DOGS ATTENDED BY THE BIRTH CONTROL PROJECT FROM UNIVERSIDADE ESTADUAL DE LONDRINA, PARANÁ, BRAZIL

CALDART, E.T.; CONSTANTINO, C.; PASQUALI, A.K.S.; DIAS, R.C.F.; MASCARENHAS, N.M.F.; FREIRE, R.L.; NAVARRO, I.T. UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL.

Keyword:seroprevalence; leishmaniasis; dog

Abstract: American cutaneous leishmaniasis (ACL) is a zoonotic disease that is endemic in many parts of Latin America. Infected dogs play a role in transmission of the parasite to humans because they appear as a link between wild and peridomestic cycle of leishmaniasis and may become an amplifier of the disease in the ecosystem. The aim of this work was to determine the prevalence of anti-L. amazonensis IgG antibodies and associated factors to infection in dogs attended by The Birth Control Project at Universidade Estadual de Londrina, Brazil, from 2004 to 2012.

Serum samples from 232 animals were submitted to Indirect Fluorescence Antibody Test (IFAT) and titers 40 were considered positives. Epidemiological surveys from 40.9% of the animals were obtained and analyzed with EpiInfo (3.4.3) statistics software. Antibodies to L. amazonensis were found in three of 232 (1.3%) animals; two of them showed a titer of 40 and one a titer of 80. The sample was consisted of: 51.5% females; 69.5% undefined breed; 25.0% aged less than 1 year and 90.3% from urban area. There was no statistical association among canine leishmaniasis occurrence and sex, breed, age, source or with variables linked to sanitation as: presence of septic tank, nearby wasteland, public garbage collection and garbage thrown on wasteland. The results show a low prevalence of anti-L. amazonensis IgG antibodies.

Previous studies have shown a prevalence of 13.9% in three different regions of Paraná State. In this study the low prevalence can be explained for the origin of the samples, because they were obtained from healthy animals that were undergoing surgical castration. The absence of any statistic significance can be explained for the low number of positive dogs.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [417] P 717 - PHLEBOTOMUS PAPATASI EXPOSURE CROSS-PROTECTS MICE AGAINST LEISHMANIA MAJOR CO-INOCULATED WITH PHLEBOTOMUS DUBOSCQI SALIVA KRATOCHVÍLOVÁ, T.; VOTÝPKA, J.; VOLF, P.; ROHOUŠOVÁ, I.

DEPARTMENT OF PARASITOLOGY, FACULTY OF SCIENCE, CHARLES UNIVERSITY IN PRAGUE, PRAGUE, CZECH REPUBLIC.

Keyword:immunology, sand fly, saliva; leishmania, macrophages; nitric oxide, arginase, cytokines

Abstract: Leishmania parasites are inoculated into the host skin together with vector saliva, which plays the key role in the establishment of Leishmania infection. In a naive host saliva supresses the host immune response assisting Leishmania to establish the infection, while in a repeatedly bitten host it elicits a protective immune response. Selected salivary proteins have already been successfully tested as candidates for the transmission-blocking vaccine. However, there are more than 30 sand fly species proven as Leishmania vectors, all differing in composition of saliva. Therefore, the specificity of the protective immunity could negatively affect the development of general vector-based vaccine.

The main aim of our experiments was to examine the possibility of cross-protective effect between saliva of two closely related species, P. papatasi and P. duboscqi, the natural vectors of Leishmania major. BALB/c mice were twice exposed to P. papatasi bites and then challenged with L. major together with P. papatasi or P. duboscqi saliva. As shown previously, mice immunized by P. papatasi were protected against Leishmania infection when challenged with homologous saliva. This was reflected by significantly smaller ear lesion size, lower parasite load in the ear dermis and in draining lymph nodes, and lower level of anti-L. major IgG.

Moreover, we also determined the effect of sand fly saliva on macrophages, the key cells in the infection control. The protective effect of immunized group was reflected in increased production of TNF- , while the production of IL-10 and nitric oxide was reduced. For the first time, we showed the cross protection in a P. papatasi-immunized mice challenged in the presence of P. duboscqi saliva. When compared to P. papatasi-challenged group, there was no difference in Leishmania lesion size, parasite load in the infected ear and draining lymph node, production of NO, TNF- , IL-12, or in the levels of anti-L. major IgG. The protective effect was demonstrated by significantly decreased lesion size and by reduction of parasite load in the draining lymph node, however, the levels of anti-L. major IgG and the production of NO and cytokines remained comparable to nonimmunized group.

In conclusion, the cross-protectivity effect between both vectors of L. major suggests that the same saliva-based vaccine could be applicable in several L. major foci.

- EPIDEMIOLOGY [422] P 718 - EPIDEMIOLOGIC ASPECTS OF AMERICAN CUTANEOUS LEISHMANIASIS IN THE CITY OF PORTO VELHO WITHIN THE PERIOD FROM 2007 TO 2012 – RO SILVA JÚNIOR, C.F.; DILL, D.C.S.; SOUZA, P.R.; GUEDES, R.P.O.; CASTILHO, W.S. UNIVERSIDADE FEDERAL DE RONDÔNIA, PORTO VELHO, RO, BRAZIL.

Keyword:leishmaniasis; epidemiology; reporting

Abstract: Introduction: Leishmaniasis is an antropozoonoses and considered a major public health problem in Brazil and worldwide. Cutaneous leishmaniasis presents classically as a disease that manifests itself in two main forms: cutaneous leishmaniasis (CL) and mucosal leishmaniasis or (ML) mucocutaneous (LMC).

Methodology: The research is a qualitative and retrospective study. The study population included patients with leishmaniasis in the city of Porto Velho, Rondônia. Data were obtained from the Municipal Health Office- SEMUSA, which are related to cases of leishmaniasis by age group, sex, clinical presentation, type of input and evolution, reported within the period 2007 to 2012.

Objectives: to describe the epidemiology of leishmaniasis in the city of Porto Velho within the period from 2007 to 2012 – RO. Results and discussion: within the period 2007 to 2012 is reported 588 cases of leishmaniasis in the city of Porto Velho and of this total, 93% were recorded as new cases and the remainder as relapses. Of this total, 394 cases (67%) were recorded within age group 15 to 44 years, 43 (7.3%) within the period 0-14 years, 87 (14.7%) within 45-54 years and 63 (11%) above 55 years. Regarding to sex, we observed a predominance of males, with 87% of cases. According clinical form, were reported 86% of cases of CL against 14% of cases of ML. Regarding the evolution of disease over than 23% had their treatment outcomes ignored, 75% evolved to healing, the treatment dropout rate was about 1% and 1% remainder included one death by CL, in 2011, one death from other causes in 2009, and three transfer cases over the study period.

Conclusion: in conclusion, it was observed that even in the face of so many reported cases of the disease there is still much underreporting. Most cases occur in individuals in the productive age group, mainly within 15 and 44 years old, and still predominant in males. Leishmaniasis in its cutaneous form is the most prevalent clinical presentation equivalent to 86% of cases in the

study population. This study demonstrated a high cure rate (75% of cases), reflecting positively on the quality of life of patients, thus justifying active search for cases and early treatment of disease.

- RESERVOIRS [424] P 719 - HEMATOLOGICAL AND BONE MARROW CHANGES IN SEMI-DOMICILED DOGS NATURALLY INFECTED WITH LEISHMANIA INFANTUM ALMEIDA, V.A.1; MOURA-NETO, J.P.2; ARAÚJO, I.B.3; DOS-SANTOS, W.L.C.1 1.FUNDAÇÃO OSWALDO CRUZ, CENTRO DE PESQUISAS GONÇALO MONIZ, SALVADOR, BA, BRAZIL; 2.UNIVERSIDADE FEDERAL DO AMAZONAS, MANAUS, AM, BRAZIL; 3.UNIVERSIDADE FEDERAL DA BAHIA, SALVADOR, BA, BRAZIL.

Keyword:canine visceral leishmaniasis; bone marrow; natural infection

Abstract: Introduction: Bone marrow function is frequently impaired by infections with L. infantum and other pathogens. These infections are common among semi-domiciled dogs of Brazil.

Objectives: To study the haematological changes in peripheral blood and morphological changes of bone marrow present in semi-domiciled dogs naturally infected with L. infantum. Methods: 50 stray dogs from Jequié (BA), an endemic area of visceral leishmaniasis were subjected to clinical examination, hemogram, laboratorial tests for detecting Leishmania infection (PCR, ELISA, and culture of splenic aspirate). They were subject to necropsy and tissue samples were collected. Hematoxylin-eosin, PAS and Giemsa-stained bone marrow sections were examined considering the following parameters: cellularity, cell populations hypoplasia or hyperplasia and other histological alterations. The animals were classified according to the number of clinical signs of visceral leishmaniasis (weight loss, conjunctivitis, pale mucous membranes, dermatitis, alopecia, onychogryphosis, lymphadenopathy and splenomegaly) as asymptomatic (up to 1 sign) oligosymptomatic (2-3 signs) or polysymptomatic (more than 3 signs). Results: Leishmania infection was detected in 50 (100%) by PCR, in 41 (82%) dogs by ELISA and in 15 (30%) by spleen culture. The most frequent haematological changes observed in peripheral blood were anemia (72,3%), leukocytosis (34,0%) thrombocytopenia (66,0%). In bone marrow there were granulocytic hyperplasia (98,0%), plasmocytosis (96,0%), megakaryocytic hyperplasia (62,0%), histiocytosis (60,0%), sinusoidal congestion (36,0%), interstitial hemorrhage and emperipolesis (30,0%), and interstitial edema (8,0%). Peripheral blood leukocytosis (61,5%) and bone marrow histiocytosis (93,3%) were more frequent in animals with, than in animals without (23,5%, P=0.037 and 45,7%, P=0.002, respectively) positive splenic culture. Conclusions: Bone marrow changes are frequent in semi-domiciled animals. However, active infection by L. infantum in these animals is associated with increased peripheral white blood cell counts and with an increased number of macrophages in the bone marrow.

CYTED-RIMLEV, FIOCRUZ-PAPES V, CNPq. valter.almeida@aluno.bahia.fiocruz.br - EPIDEMIOLOGY [436] P 720 - URBAN LAND USE CHARACTERISTICS ASSOCIATED WITH THE PREVALENCE OF CANINE LEISHMANIA CHAGASI INFECTION IN TERESINA, BRAZIL: A STUDY BASED ON REMOTE SENSING FERNANDES, A.B.1; WERNECK, G.L.1; ALMEIDA, A.S.2; CRUZ, M.D.S.P.E.3; SILVA, J.P.4

1.UNIVERSIDADE FEDERAL DO RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 2.UNIVERSIDADE DO ESTADO DO RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 3.UNIVERSIDADE FEDERAL DO PIAUÍ, TERESINA, PI, BRAZIL; 4.GERÊNCIA DE CONTROLE DE ZOONOSES/FMS, TERESINA, PI, BRAZIL.

Keyword:canine visceral leishmaniasis; risk factors; remote sensing

Abstract: The objective of this study was to examine the association between land use characteristics assessed through remote sensing and the prevalence of canine Leishmania chagasi infection in an endemic area for visceral leishmaniasis. This is a case-control study based on the results of a routine seroepidemiological among domestic dogs performed in 2007 in the Angelim neighborhood, located in the city of Teresina, state of Piaui, Brazil. Canine infection was evaluated by means of the of indirect immunofluorescence antibody test. All dwellings in which at least one seropositive dog was detected were considered cases, and controls were a random sample of dwellings in which only seronegative dogs were identified. Two satellite scenes (CBERS 2/CCD from July/2006 with 20 meters spatial resolution and a CBERS 2B/HRC from October/2008 de 2008 with 2,5 meters spatial resolution), were used to describe land use features in pixels of 625m2 over the whole study area. Associations between variables were expressed as odds ratios (OR) and their respective 95% confidence intervals (95%CI) estimated using multivariate logistic regression. After adjustment for socioeconomic variables, the odds of infection was two times higher for houses located in pixels with 25m2 of the area covered by residential structures with little vegetation as compared to those located in areas in which this type of land use covered 600m2. Similarly, houses located in pixels with less than half of the area covered by residential structures with abundant vegetation showed odds of infection 60% smaller as compared to those located in areas almost totally covered by this type of land use.

The results of this study reinforce the whole of proximity to vegetation as a potential risk factor for canine Leishmania chagasi infection. In urban areas the interplay between the size of the population, as indicated by residential structures, and the area covered by vegetation should be better investigated in future studies. The identification of land use features associated to canine Leishmania chagasi infection might be an useful tool for identifying areas at higher risk for human visceral leishmaniasis, and help orient the implementation of focused interventions.

- CELLULAR AND MOLECULAR BIOLOGY [455] P 721 - COINFECTION OF EHRLICHIA CANIS AND LEISHMANIA INFANTUM CHAGASI IN CANINE MACROPHAGES (DH82 CELLS).

CARVALHO, R.M.A.; SILVA, K.R.; BARRAL, M.A.D.P.; BARRAL-NETTO, M. CENTRO DE PESQUISA GONĂ&#x2021;ALO MONIZ-FIOCRUZ/BA, SALVADOR, BA, BRAZIL.

Keyword:ehrlichia canis; leishmania infantum chagasi; coinfection

Abstract: Canine leishmaniosis and canine ehrlichiosis have common endemic areas, the same periods of transmission and share several manifestations of their clinical pattern (anorexia, fever, lymphadenomegaly, splenomegaly, bleeding) and laboratory tests profile (thrombocytopenia and pancytonenia). Both are infectious diseases caused by intracellular parasites, Leishmania infantum chagasi and Ehrlichia canis, respectively. Co-infection in dogs has been reported, but there are few studies exploring its characteristics. This report aims to understand the mutual influence of these parasites during in vitro coinfection. For this, we cultivated DH82 cells (a canine macrophage cell line) in DMEM medium (107 cells per ml) with 10% calf serum for 10 days, and then infected them with L. infantum chagasi (108 parasites/ml), or with E. canis (107 infected cells per ml) or both at the same time. Infection was evaluated by optical microscope at different times points (20h, 28h, 48h, 4d, 5d, 7d, 12d and18d). In DH82- Leishmania culture, the first four days was observed 38% of infection than the infections was decreasing until 5% in day 18. In DH82-Ehrlichia culture the infection increased from 3% in the first three days until 90% in day 18.In coinfected culture a low infection was observed of both parasites, 6% of Erlichia canis and 2% of Leishmania infantum chagasi until day 5, when the Ehrlichia begins to be more prevalent, until have 90% of infection in day 18 and infection for Leishmania was not observed.

We conclude that E. canis is able to infected DH82 and maintain this infection, while L. chagasi is able to infected DH82 cells but not sustain the infection.

- EPIDEMIOLOGY [460] P 722 - DETECTION AND FOLLOW-UP OF LEISHMANIA INFANTUM INFECTION IN ASYMPTOMATIC BLOOD DONORS FROM BALEARIC ISLANDS (SPAIN) FISA, R.1; RIERA, C.1; GUILLEN, C.1; INIESTA, L.1; BULA, I.1; CANCINO, B.1; GIRONALLOBERA, E.2; SEDEÑO, M.2; JIMENEZ-MARCO, T.2 1.DEPT MICROBIOLOGIA I PARASITOLOGIA SANITÀRIES, FACULTAT DE FARMÀCIA, UNIVERSITAT DE BARCELONA, BARCELONA, SPAIN; 2.FUNDACIÓ BANC DE SANG I TEIXIT DE LES ILLES BALEARS, PALMA DE MALLORCA, SPAIN.

Keyword:leishmania infantum; asymptomatic infection; blood donor

Abstract: The existence of asymptomatic Leishmania infantum carriers is common in the European Mediterranean basin. These asymptomatic carriers could represent a challenge for transfusion safety in our region. The aim of this study was to detect asymptomatic carries and to follow-up the asymptomatic infection in blood donors from endemic areas located in Balearic Islands, as an approach to improve our knowledge about the L. infantum transfusion-transmission risk in our region.

The population study consisted in 858 blood donors who lived in the Balearic Islands and had no history of leishmaniasis, recruited from October 2008 to December 2012. Antibody response was analyzed by performing Western Blot (WB) in serum samples from all blood donors. The presence of Leishmania spp. DNA was analyzed by amplification of kinetoplast DNA by realtime PCR (qPCR) in 175 peripheral blood. Each amplification was performed in triplicate and qPCR was considered positive when the threshold cycle (tC) was <40.

Among all samples studied 63 samples were positive by WB testing and 20 samples were positive by qPCR. The prevalence rates were 7.3% (5.8-9.3%) by WB and 11.4% (7.5-17.1%) by qPCR. When detectable, the parasitemia ranged from 0.01 parasites/mL to 1 parasite/mL. A subgroup of 15 blood donors that were identified as asymptomatic carriers, with positive results by serology and/or by qPCR, was followed-up over the study period. The follow-up showed that: a donor had repeated positive WB and peripheral qPCR on two occasions; two donors presented detectable antibody and negative qPCR and twelve donors had L. infantum detected in peripheral blood by qPCR at least once. However, seven of them did not present antibody response during their follow-up period.

Accordingly this study, Balearic Islands as an endemic region, shows persistently a significant proportion of L. infantum asymptomatic carriers over time. Some donors have the parasite circulating in peripheral blood but undetectable antibody levels and others have persistently detectable antibodies without circulating parasites. In consequence, a blood donor screening test based solely on antibody detection does not seem to be sufficient to identify asymptomatic carriers. Interestingly, qPCR enables adequate monitoring of Leishmania spp. indicating that the presence of the parasite in the bloodstream of asymptomatic carriers is episodic and at low density.

- EPIDEMIOLOGY [469] P 723 - LOCAL EPIDEMIOLOGICAL INDICATORS AND THE SURVEILLANCE OF VISCERAL LEISHMANIASIS IN HIGH TRANSMISSION AREAS BARBOSA, D.S.; BELO, V.S.; WERNECK, G.L. FIOCRUZ, RJ, RJ, BRAZIL.

Keyword:epidemiological indicators; medical geography; visceral leishmaniasis

Abstract: The definition and use of local epidemiological indicators in the health services for identification of priority areas for surveillance purposes and implementation of control measures against visceral leishmaniasis (VL) are of fundamental importance in the context of disease spread. We aimed to explore the use of local epidemiological data about VL for monitoring disease spread and for supporting surveillance and control measures against the disease in high transmission areas. We performed an ecological study using VL cases reported in the city of São Luís, the capital of the state of Maranhão in northeast Brazil, a high transmission area of the disease. We evaluated the spatial structure of the incidence rates of human VL and of the mean numbers of human cases occurring in the 355 neighborhoods of the city (aggregated into 203 units of analysis) between 1999 and 2007. To explore the changes in the spatial distribution of the disease, maps were construct for three periods: 1999-2001, 2002-2004, 2005-2007. To avoid instability in incidence rates calculation due to small population in the units of analysis, we used a local Bayesian empirical correction. Using the mean number of human cases, the indicator suggested by the Brazilian Ministry of Health to classify the level of transmission, we found that the spatial distribution of disease in the units of analysis, showed low values that can mask the real situation of the disease in the municipality. Using this indicator makes it difficult to identify critical areas for intervention, because the fragmented population in smaller units of analysis within the city decreases the possibility of occurrence of enough human cases in order to assess the dynamics of disease spread. The spatial distribution of the disease can be better captured when we use the incidence of human disease as the local indicator, offering a more realistic background of the situation. Results of this study suggest that the definition of which epidemiological indicator should be used at the local level needs to be revised. A better definition of the priority areas for implementing control actions against the disease should take into account not only the local level of incidence, but should also include information about the surrounding areas, in particular those that appear to have a strong influence on the spread of visceral leishmaniasis.

- EPIDEMIOLOGY [474] P 724 - MOLECULAR EPIDEMIOLOGY OF CUTANEOUS LEISHMANIASIS IN LIBYA 1995-2012 AMRO, A.1; AL-DWIBE, H.2; GASHOUT, A.2; ALAM, M.Z.3; ANNAJAR, B.2; HAMARSHEH, O.4; SCHÖNIAN, G.5 1.FACULTY OF PHARMACY, AL-QUDS UNIVERSITY, JERUSALEM, PALESTINE, JORDAN; 2.LIBYAN NATIONAL CENTRE FOR INFECTIOUS DISEASES AND CONTROL, TRIPOLI, LIBIA; 3.DEPARTMENT OF PARASITOLOGY, FACULTY OF VETERINARY SCIENCE, BANGLADESH AGRICULTURAL UNIVERSITY,, MYMENSINGH, BANGLADESH; 4.DEPARTMENT OF BIOLOGICAL SCIENCES, FACULTY OF SCIENCE AND TECHNOLOGY, AL- QUDS UNIVERSITY, JERUSALEM, PALESTINE, JORDAN; 5.INSTITUTE OF MICROBIOLOGY AND HYGIENE, CHARITÉ UNIVERSITY MEDICINE BERLIN, BERLIN, GERMANY.

Keyword:keywords. libya, cutaneous leishmaniasis, leishmania tropica, .; leishmania major; internal transcribed spacer 1 its1

Abstract: Cutaneous leishmaniasis (CL) has been prevalent for many decades in Libya. Disease epidemiology and characterization of the causative leishmanial species were not investigated in

depth. The objective of this study was to investigate clinical and epidemiological features of CL outbreaks in Libya including molecular identification of parasites, the geographical distribution of cases and possible scenarios of parasite transmission and life cycle.

We studied 665 patients, who came to the Libyan National Centre for Infectious Diseases and Control, Tripoli, Libya during 1995-2012. These cases came from 49 areas distributed in 12 districts in north-west Libya. The patients’ age ranged from 4 months to 89 years (median age 26 years), and 53% of the cases were males and 47% females. Skin scrapings spotted on glass slides were collected for molecular identification of causative agent. The ribosomal internal transcribed spacer 1 (ITS1) was amplified and subsequently characterized by restriction fragment length polymorphism (RFLP) analysis. In total, 195 samples were successfully identified of which 148 (75.9%) were Leishmania major, and 47 (24.1%) were L. tropica. CL cases infected with L. major were found in all CL areas whereas L. tropica cases came mainly from Al Jabal Al Gharbi (46.4%), Misrata (17.8%) and Tarhuna districts (10.7%).

A trend of seasonality was noticed for the infections with L. major which showed a clear peaked between November and January, but was less pronounced for infections by L. tropica.

The recent armed conflict in Libya has lead to massive human migrations and resulted in inadequate functioning of the primary health care system, inadequate surveillance, and the complete interruption of control programs. Quantification and containment of these risk factors are under investigation.

This is the first molecular study on CL in Libya. The epidemiological patterns in the different foci were the same as in other Mediterranean foci of CL where disease was caused by L. tropica and L. major.

- EPIDEMIOLOGY [481] P 725 - ECOEPIDEMIOLOGY OF AMERICAN VISCERAL LEISHMANIASIS IN STATE OF TOCANTINS, BRAZIL, AND FACTORS ASSOCIATED WITH VECTOR LUZOMYIA (L.) LONGIPALPIS (LUTZ & NEIVA, 1912) (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) AFONSO, M.M.S.1; CHAVES, S.A.M.2; MAGALHÃES, M.A.F.3; CARRIJO, R.S.G.G.3; AZEVEDO, C.G.4; BOU HAYA, J.V.5; RANGEL, E.F.5 1.INSTITUTO OSWALDO CRUZ / ESCOLA NACIONA DE SAÚDE PÚBLICA SÉRGIO AROUCA / FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 2.ESCOLA NACIONAL DE SAÚDE PÚBLICA SÉRGIO AROUCA / FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.INSTITUTO DE COMUNICAÇÃO E INFORMAÇÃO CIENTÍFICA E TECNOLÓGICA EM SAÚDE / FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 4.SECRETARIA DE SAÚDE DO ESTADO DE TOCANTINS, PALMAS, TO, BRAZIL; 5.INSTITUTO OSWALDO CRUZ / FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:lutzomyia (l.) longipalpis ; lane use; american visceral leishmaniasis

Abstract: In Brazil, Leishmaniases are serious public health problems and the geographical expansion has enabled the diseases to become established in urban areas of large cities. This scenario occurs in the state of Tocantins (TO), which in recent years has shown high rates of American Visceral Leishmaniasis (AVL). The aim of this study was to correlate the classes of land use in the presence of Lutzomyia (Lutzomyia) longipalpis vector, the records of human cases (2001 to 2011) and to analyze the AVL transmission stratification (2004 to 2011). Data for classes of land use were available from the Federal University of Goiás and the Brazilian Institute of Geography and Statistics (32 levels of phyto-ecological formation). The records of human cases of AVL were obtained in the Brazilian Notifiable Diseases Information System database and for

the identification of municipalities with the presence of the vector, were used bibliographic consultations. The stratification data for AVL transmission was assigned by the Tocantins Department of Health. Among 139 municipalities, there were reported cases of AVL in 120 and L. (L) longipalpis was registered in 26. Regarding the presence of human cases of AVL, there was a positive correlation with wooded savannah (p=0.203), open rainforest submontane (p=0.248), secondary vegetation (p=0.202), rainforest-savannah ecotone (p=0.259), grazing pasture (p=0.241) and grassy-wooded savannah (p=0.175); negative correlation was observed with cultivated pasture (p=-0.196), wooded savannah gallery forest (p=-0.278) and savannah parkland with gallery forest (p=-0.192). Lutzomyia (L.) longipalpis was associated with all classes of land use, a fact that confirms its expansion and adaptation to different environments, including antropized areas. Regarding to epidemiological analysis of stratification of AVL, of the total number of municipalities (139), 29 of them maintained the same transmission profile: 22, no transmission, 3, sporadic transmission and 4, intense transmission. It is interesting to note that 55 municipalities had an increase in the number of human cases during recent years. In 2010, 12 municipalities demonstrated intense transmission and in 2011 this number rose to 17.

The study discusses the AVL in Tocantins in light of expansion as a result of environmental impacts and adaptation of vector L. (L.) longipalpis to various habitats. Such evidence must be considered in the planning of epidemiological surveillance measures.

Supported by: Instituto Oswaldo Cruz/FIOCRUZ; CAPES; FAPERJ - Brasil

- EPIDEMIOLOGY [487] P 726 - SEROPREVALENCE OF LEISHMANIA INFANTUM AND TOXOPLASMA GONDII IN HORSES FROM NORTHERN PORTUGAL LOPES, A.P.1; SOUSA, S.2; RIBEIRO, A.J.3; MAIA, J.M.3; SILVESTRE, R.2; COTOVIO, M.1; SCHALLIG, H.4; CORDEIRO-DA-SILVA, A.2; CARDOSO, L.1

1.DEPARTMENT OF VETERINARY SCIENCES AND CECAV, ECAV, UNIVERSITY OF TRÁS-OS-MONTES E ALTO DOURO (UTAD), VILA REAL, PORTUGAL; 2.PARASITE DISEASE GROUP, INSTITUTO DE BIOLOGIA MOLECULAR E CELULAR (IBMC), UNIVERSIDADE DO PORTO, OPORTO, PORTUGAL; 3.VETERINARY HOSPITAL, UTAD, VILA REAL, PORTUGAL; 4.KONINKLIJK INSTITUUT VOOR DE TROPEN (KIT), ROYAL TROPICAL INSTITUTE, DEPARTMENT OF PARASITOLOGY, AMSTERDAM, NETHERLANDS.

Keyword:leishmania infantum, horses; portugal, dat; toxoplasma gondii, mat

Abstract: Leishmaniosis caused by Leishmania infantum (syn. Leishmania chagasi) is endemic in the Mediterranean basin (including Portugal), the Americas and Asia. Dogs are the main reservoir of the parasite, which is transmitted among animals and to human beings by phlebotomine sand flies. In Europe, cases of presumed autochthonous equine cutaneous leishmaniosis have been sporadically reported from Germany, Spain, Portugal and Switzerland. Toxoplasmosis can be the cause of morbidity and mortality in congenitally infected children and immunocompromised people. Cats and wild felids are the only recognized definitive hosts of Toxoplasma gondii, while a large variety of homeothermic animals, including humans and horses, serve as intermediate hosts. Toxoplasma gondii infection in horses is often subclinical, although fatal illness may develop. As herbivores, horses can serve as sentinels of environmental contamination with T. gondii oocysts and, additionally, the consumption of improperly cooked infected horse meat can be a health risk to humans. Serological tests to detect parasite-specific antibodies are useful for epidemiologic studies of infection. Between November 2008 and July 2010, plasma samples were obtained from 173 horses living in northern Portugal. The direct agglutination test (DAT, KIT) was used for titration of IgG antibodies specific to Leishmania.

Samples were tested for IgG to T. gondii with a direct microagglutination commercial kit (ToxoScreen DA®, bioMérieux). Thirty horses were found seropositive to at least one of the agents

(17.3%; 95% CI: 12.0-23.8). The seven horses seropositive to L. infantum had titers of 200 (n = 5), 800 (n = 1) and 1,600 (n = 1). The 23 horses seropositive to T. gondii (13.3%; 95% CI: 8.6-19.3) had titers of 20 (n = 13), 40 (n = 5), 80 (n = 3) and 160 (n = 2). The age of seropositive animals ranged from 1 to 18 years for L. infantum and from 3 to 25 years for T. gondii. A statistically significant difference was found between the seroprevalence of L. infantum and T. gondii, but with none of the horses being simultaneously seropositive to L. infantum and T. gondii, which rules out cross reactions. Horsesâ&#x20AC;&#x2122; gender, type of housing, ability and clinical status (apparently healthy or sick) were not associated with seropositivity to L. infantum or T. gondii. Results of the present study indicate that horses from northern Portugal are exposed to and may be infected with L. infantum and T. gondii.

- EPIDEMIOLOGY [491] P 727 - CUTANEOUS LEISMANIASIS TRUJILLO VENEZUELA ROJAS, E.M.; SCORZA, J.V. INSTITUTO EXPERIMENTAL, TRUJILLO, VENEZUELA.

Keyword:leishmania (v) braziliensis; ecoepidemiology; urban incidence

Abstract: The natural history of this enzootia considers its occurrence in wild environment and for the man's presence it becomes an occupational illness of lumberjacks and hunters. With the intention of describing a new local epidemic condition for leishmaniasis, we intend to analyze and to relate some parameters that identify the clinical case and the area it inhabits and to compare the towns between them to determine transmission risk factors in urban populations.

We sampled selectively 10% of the total population of the city to obtain epidemic information about suffering or not leishmaniasis and to update the prevalence in the city. Some social factors and migration were also evaluated. This information also gave environmental and entomological punctual information The sample of 4000 people was analyzed by families with and without cases of leishmaniasis, it was diagnosed and clinically treated when it was necessary. This sample was considered as macro universe for the statistical populational analysis of the localities. We collected a sample of blood for immunologic and molecular tests. We evaluated the clinical antecedents of former and recent cases in each clinical history when these were treated in the Instituto J.W.T. The analysis to macro level (P>95%) allowed to group the inhabited localities of the sample and they were contrasted to each other to elaborate a dendogram of epidemic similarity based on the vectorial transmission rate, risk factor determined as more important.

Geographically, the populations that inhabit areas with little direct solar illumination independently of the housing type presented more cases (60%) that those that inhabit the areas better illuminated (40%). It was not observed significant differences by altitude for the number of cases located among 790-980 masl. We confirmed that in more than 1300 masl they didn't have Leishmania transmission. The analysis of similarity of a total of 54 compared localities separated three big populations groups that coincide with the geographical location. With molecular tests (PCR) we confirmed in 95% of the cases studied Leishmania (Viannia) braziliensis, as causal agent of the leishmaniasis in the study area. The response to the treatment with diverse systemic therapies for clinic cure demonstrated in the sample that late mucosal engagement doesn't exist. The analysis with PCR-hibrydation demonstrated that the type of treatment neither the time post-cure affects the serological detection of DNA of Leishmania (V) braziliensis. Complement diagnosis ELISA technique didn't evidence seroconvertion in the postcure analyzed cases.

- CONTROL PROGRAMS [492] P 728 - SPATIAL DISTRIBUTION OF VISCERAL LEISHMANIASIS IN THE STATE OF BAHIA, BRAZIL: IMPLICATION FOR ECONOMIC DEVELOPMENT AND CHALLENGES FOR ITS PREVENTION. JÚNIOR, E.R.1; CASTRO, J.M.1; PATEL, B.N.2 1.DIRETORIA DE VIGILÂNCIA EPIDEMIOLÓGICA DA SECRETARIA DE SAÚDE DO ESTADO DA BAHIA, SALVADOR, BA, BRAZIL; 2.UNIVERSIDADE ESTADUAL DE FEIRA DE SANTANA, FEIRA DE SANTANA, BA, BRAZIL.

Keyword:epidemiology; bahia; visceral leishmaniasis

Abstract: Visceral leishmaniasis (VL) is a anthropozoonosis disease caused by an obligate intracellular protozoa of the genus Leishmania. It is considered a serious infectious disease, which may present high rate of mortality in endemic areas. The objective of this study is to identify the spatial distribution of LV in Bahia Method: For this purpose, the database” SINAN” of the State Department of Health of Bahia (SESAB), from 2009 to 2011 were consulted.. The degree of LV incidence in the municipalities were classified in to four categories: ‘sporadic’ (meaning less than 2.4 cases of LV during the past three years), “moderate” ( equal to greater than 2.4 cases but less than 4.4 cases of LV), “severe” ( equal to or greater than 4.4 cases of LV) and “silent” (have no reported cases). In Bahia has humid, semi humid and dry climatic regions with different types of vegetation. Results: The data demonstrated a large variation in the incidence of VL in Bahia. From a total of all the 417 municipalities of Bahia, included in the study , 218 (52.27%) presented positive new cases of VL. In 199 (47.72%) of the municipalities no cases of LV were reported, where as 17 (7.79%) of the municipalities were classified in the “ “intense” degree of infestation. In 23 (10.55%) municipalities the LV infection was in the “ moderate “ class. The “sporadic” category of LV infestation was found in 137 (62.84%) of the municipalities. The 17 municipalities with severe infestation were Guanambi, Juazeiro, Salinas da Margarida, Iraquara, Jequié, Cafarnaum, Canarana, Feira de Santana, Irecê, Uibaí, Cabaceiras do Paraguaçu, America Dourada, Sento Sé, Boquira, Camaçari, Barro Alto and Euclides da Cunha. Discussion: We observed that four municipalities, classified as "intense" LV infestation, (Guanambi, Juazeiro, Camaçari and Feira de Santana), present great potential for economic development. These municipalities are undergoing developmental processes that lead to deforestation and the invasion of green areas, followed by intensive irrigated agricultural industry. This situation profoundly alter the ecosystem, favoring the breeding of the leismania vectors and consequent possible transmission of VL, to humans and animals. Conclusion: Taking into consideration the inevitable need for the socio-economic development of Bahia, the current and future challenges are to reconcile this development, taking into account the needs of the public health issues, especially of the visceral Leishmaniasis. There is an urgent necessity for broad and harmonious discussions between the various segments at multi-institutional levels and multidisciplinary segments in Bahia. Public managers( at the state and municipal level) and managers of the agricultural industry, agribusiness, tourism industry, civil construction sector and members of the organized civil society must jointly discuss and develop plans for a sustainable environmental, economic and social development in Bahia.

- EPIDEMIOLOGY [496] P 729 - EPIDEMIOLOGY OF VISCERAL LEISHMANIASIS IN THE CITY OF CAXIAS – MA NETO DA SILVA, M.A.C.1; JARDIM FILHO, W.R.2; RIBEIRO, Y.J.S.1; COSTA, E.M.1; CASTELO BRANCO, R.C.1; MUNIZ, C.R.1; DE JESUS, E.A.1; MONTEIRO, E.L.O.1; JUCA, A.A.1; CRUZ, A.S.1; RODRIGUES, G.M.1; DA SILVA, I.S.2; BEZERRA, G.F.B.1; NASCIMENTO, M.D.D.S.B.1 1.UNIVERSIDADE FEDERAL DO MARANHÃO, SÃO LUÍS, MA, BRAZIL; 2.UNIVERSIDADE ESTADUAL DO MARANHÃO, CAXIAS, MA, BRAZIL.

Keyword:visceral leishmaniasis; epidemiology; caxias - ma

Abstract: Introduction: In Brazil, visceral leishmaniasis (VL) or kala-azar is endemic neotropical, mainly rural, with trends towards urbanization. The etiological agent of VL is Leishmania chagasi, the protozoan intracellular mononuclear phagocytic system. The LV is widely distributed throughout the world. The estimated risk population of 350 million people, currently endemic in 88 countries. In Latin America, Brazil is responsible for 90% of reported cases, 56% in the Northeast. Between 1984 and 2002, 66% of cases occurred in the states of Maranhão, Piauí, Ceará and Bahia. The VL in Brazil affects mainly children, with higher prevalence in males and with the main reservoir Dog From 2006, in Brazil, every case of VL is notifiable, and the data recorded in the Information System Diseases Notification - SINAN. The VT has interrelationships geographic, climatic, social and cultural, which diversifies its clinical manifestations in different regions of the country, as well as being risk factors for the occurrence of the disease. Material and Methods: The study was based on retrospective analysis, cross sectional, descriptive, and quantitative nature documentary of 180 cases of patients diagnosed with visceral leishmaniasis in the period 2007 to 2011, in the city of Caxias - MA. Data were provided by the Department of Epidemiological Surveillance. Were analyzed the following variables: age, sex, race, neighborhood, co-infection with HIV, area of residence, type of entry, education, relationship with work and evolution. Results: We observed that the total of 180 cases (100%), 70% were aged 9 years old, 108 cases were male (60%) and 72 cases were female (40%). As for race, 80.6% were mixed. The neighborhood undertakers was most prevalent (11.25%). Only 3.33% had HIV co-infection. Regarding the area of residence, 88.4% of cases occurred in urban areas, 96.6% were new cases, 43% of cases had incomplete primary education, 3% of the cases were related to work and 92.3% were cured.Conclusions: Visceral leishmaniasis is endemic in the city of Caxias. The council has shown a reduction in the number of cases of illness due to actions of the Municipal Center for Health Education (NMES) with lectures and action of health workers in educating the population, however it still needs more government support in the implementation of most effective measures for the control of VL. - CLINICAL LEISHMANIASIS [498] P 730 - VISCERAL LEISHMANIASIS: LABORATORY AND CLINICAL ASPECTS IN THE CITY OF CAXIAS – MA NETO DA SILVA, M.A.C.1; JARDIM FILHO, W.R.2; RIBEIRO, Y.J.S.1; COSTA, E.M.1; CASTELO BRANCO, R.C.1; MUNIZ, C.R.1; DE JESUS, E.A.1; MONTEIRO, E.L.O.1; JUCA, A.A.1; CRUZ, A.S.1; RODRIGUES, G.M.1; DA SILVA, I.S.2; BEZERRA, G.F.B.1; NASCIMENTO, M.D.D.S.B.1 1.UNIVERSIDADE FEDERAL DO MARANHÃO, SÃO LUÍS, MA, BRAZIL; 2.UNIVERSIDADE ESTADUAL DO MARANHÃO, CAXIAS, MA, BRAZIL.

Keyword:visceral leishmaniasis; laboratory and clinic; caxias - ma

Abstract: Introduction: Visceral Leishmaniasis (VL) is an anthropozoonosis caused by a protozoan Trypnosomatidae family, genus and subgenus Leishmania and specie chagasi-L. (L.) chagasi.

VL is endemic in 88 countries, with 90% of cases occur in Brazil, especially in the Northeast. In VL, the age group below 10 years, especially the smallest of five years, is the most frequently affected. Infection by L. (L) chagasi is characterized by a fever, pallor, weight loss, increased abdominal size, hepatosplenomegaly and edema. It was also observed other clinical manifestations such as cough, diarrhea, jaundice and bleeding that hinder the differential diagnosis with other diseases, delaying the identification, which may cause the patient to death.

The diagnosis is made from clinical and epidemiological aspects, and laboratory tests such as complement fixation, indirect immunofluorescence, direct agglutination test, ELISA and DotELISA, and molecular biology techniques such as polymerase chain reaction. The gold standard is the parasitological diagnosis by aspiration of the spleen, bone marrow, liver and lymph nodes.

Material and Methods: A retrospective, descriptive study of diagnosed cases of visceral leishmaniasis in the municipality of Caxias, treated at Children's Hospital Dr. João Viana, in the period 2007 to 2011. The data analyzed were provided by the Department of Epidemiological Surveillance. Were analyzed age, sex, race, diagnostic criteria, clinical manifestations, and initial drug development. Results: Were observed that the total of 180 cases (100%), 70% were aged 9 years old, 108 cases were male (60%) and 72 cases were female (40%). As for race, 80.6% were dun. Regarding diagnostic criteria, there was prevalence of clinical and epidemiological method (85.5%). Regarding clinical manifestations, 6% had bleeding, 21.6% of infections, 51.7% hepatomegaly, splenomegaly 76.7%, pallor 79.7%, 77.9% weight loss, cough and 51.1% / or diarrhea, jaundice 24%, swelling 25.7%, 86.4% and 95 asthenia, fever 5%. The initial drug was antamonial pentavalent in 87.2% of cases. 92.3% were cured. Conclusions: Visceral leishmaniasis is an endemic disease that has many complications, especially in children. Early diagnosis and disease control should be improved VL is a public health problem and must be fought in order to minimize damage and costs.

- RESERVOIRS [500] P 731 - STUDY OF CANINE VISCERAL LEISHMANIASIS IN RECENT TRANSMISSION CITY OF MINAS GERAIS STATE, BRAZIL. SOUSA, A.M.1; MENEZES, J.A.2; MORAIS, M.H.G.1; FERREIRA, E.C.3; LIMA, F.P.4; GOMIDE, L.B.5; FONSECA, A.C.6; FILHO, V.P.6; ROCHA, A.M.S.3; FUX, B.7; SOUZA, C.M.2 1.FUNDAÇÃO EDUCACIONAL DE DIVINÓPOLIS, UNIVERSIDADE DO ESTADO DE MINAS GERAIS, DIVINÓPOLIS, MG, BRAZIL; 2.CENTRO DE PESQUISAS RENÉ RACHOU, FUNDAÇÃO OSWALDO CRUZ, BELO HORIZONTE, MG, BRAZIL; 3.UNIVERSIDADE FEDERAL DE OURO PRETO, OURO PRETO, MG, BRAZIL; 4.PREFEITURA MUNICIPAL DE FORMIGA, FORMIGA, MG, BRAZIL; 5.CLÍNICA VETERINÁRIA ESPAÇO ANIMAL, FORMIGA, MG, BRAZIL; 6.CENTRO UNIVERSITÁRIO DE FORMIGA, FORMIGA, MG, BRAZIL; 7.UNIVERSIDADE FEDERAL DO ESPÍRITO SANTO, VITÓRIA, ES, BRAZIL.

Keyword:canine visceral leishmaniasis; epidemiology; formiga

Abstract: Introduction: Visceral Leishmaniasis (VL) is an insidious disease, with high lethality rate and that urbanized in recent years. In Formiga, Minas Gerais, two autochthonous cases of human VL were reported between to 2011 and 2012, indicating the transmission occurrence in this area.

Therefore, the dog being the most important reservoir in large urban centers, the present study aimed to determinate the prevalence of canine disease in the city. Methods: In order to determine the rate of canine infection, are being conducted at random samplings of total blood of domiciled and erring dogs present in the city. This step is being carried out with the assistance of the Municipal Health Department of Formiga, the Center for the Defense of Animal Life (CODEVIDA) and by veterinary clinics in the city. The indirect immunofluorescence (IF) and enzyme linked immunosorbent assay (ELISA) were used to evaluated the prevalence of the disease, these methodology were performed at the Laboratory of Epidemiology of Federal University of Ouro Preto. Parcial Results: The Blood samples were collected from 434 dogs between June to November of 2012. Out of 8.75% (38) were positive, of which 2.53% (11) where positive only by IF and 3.68% (16) only by ELISA and 2.53% (11) of samples were positive by the two tests. The positive dogs by serology 65.8% (25), were classified as asymptomatic, corroborating in studies that most of the dogs infected do not present clinical signs of the disease. When symptomatic, most common symptoms were dermatitis, weight loss and ulcerations. Nineteen samples showed an indeterminate result by ELISA, which may represent an increase in frequency of Canine Leishmaniasis in the city, owing to possible seroconversion. Conclusions: Despite being a city with recent transmission, the rate of canine infection (8.75%) is found into levels above recommended by the World Health Organization (WHO) to implement control measures, which are 2%. Due the fact of VL be able to take on serious forms, is essential to establish preventive measures in the current scenario of the city, where the disease is not considered endemic yet. Thus, this work associated with the study of

sand flies which also comprise this research, would assist in mapping the areas of the transmission risks and the epidemiological surveillance of the disease in the city.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [501] P 732 - SPATIAL DYNAMICS OF L. LONGIPALPIS IN THE CENTRAL REGION OF CITY OF CAXIAS-MA NETO DA SILVA, M.A.C.1; DE SOUSA, M.D.G.2; RIBEIRO, Y.J.S.1; COSTA, E.M.1; CASTELO BRANCO, R.C.1; MUNIZ, C.R.1; BEZERRA, G.F.B.1; NASCIMENTO, M.D.D.S.B.1 1.UNIVERSIDADE FEDERAL DO MARANHÃO, SÃO LUÍS, MA, BRAZIL; 2.UNIVERSIDADE ESTADUAL DO MARANHÃO, CAXIAS, MA, BRAZIL.

Keyword:visceral leishmaniasis; lutzomya; caxias - ma

Abstract: Introduction: Visceral leishmaniosis (VL) is a anthropozoonosis considered one of the seven major epidemics in the world whose etiological agent is Leishmania chagasi and the vector is the female of phlebotomine Lutzomyia longipalpis, which has been found both in natural ecotopes as in rural and urban. Climatic factors, such as temperature and rainfall, affect the phlebotomine population and the knowledge of seasonality of local species important it´s in planning prevention strategies. To investigate the seasonality of L. longipalpis and the relationship between rainfall and its frequency in a central area of the city of Caxias-MA considering the relevance of this knowledge in endemia´s control. Material and Methods: We performed monthly catches, with CDC light traps in three homes at central area of Caxias Maranhão from September 2007 to June 2009. We used traps inside and outside the houses, exposed from eighteen to six hours. The specimens were identified according to the proposal of Young and Duncan. We used the Spearman test to assess possible correlations between rainfall and frequency of sand flies. Results: We found four species of phlebotomine all from the gender Lutzomyia in a total of 521 insects. The species found were L. lenti, L. evandroi, L. whitmani and L. longipalpis. This last one was the most abundant, occurring predominantly male (94.2%) and more frequently in areas surrounding homes (63.9%) throughout the study period. L. longipalpis was found in all months, except in 2007 (the first month of collection), with a higher frequency in months of rainy season or next to it. There was statistically significant with moderate positive correlations between the events studied, rainfall and monthly number of phlebotomine. Conclusions: The species L. longipalpis was the most frequent and rainfall in the study seems to be a contributing factor to the increase of its population.

- EPIDEMIOLOGY [503] P 733 - VISCERAL LEISHMANIASIS: THE EVOLUTION OF ENDEMIC IN CAXIAS - MA IN THE LAST 8 YEARS NETO DA SILVA, M.A.C.1; DE SOUSA, M.D.G.2; RIBEIRO, Y.J.S.1; COSTA, E.M.1; CASTELO BRANCO, R.C.1; MUNIZ, C.R.1; MONTEIRO, E.L.O.1; BEZERRA, G.F.B.1; NASCIMENTO, M.D.D.S.B.1 1.UNIVERSIDADE FEDERAL DO MARANHÃO, SÃO LUÍS, MA, BRAZIL; 2.UNIVERSIDADE ESTADUAL DO MARANHÃO, CAXIAS, MA, BRAZIL.

Keyword:visceral leishmaniasis; epidemiology; caxias - ma

Abstract: Introduction: Visceral leishmaniasis (VL) is a reportable disease, with clinical features of severe outcome, which remains a public health problem. This study aimed to evaluate the epidemiological, clinical, laboratory aspects and treatment of the visceral leishmaniasis,

identifying the socio-demographic aspects of the disease, raising the specific laboratory tests used for diagnosis of VL and deaths during the study period, as well as investigating the situation of canine visceral leishmaniasis in the municipality of Caxias, Maranhão. Material and Methods: We analyzed the records of the Information System for Notifiable Diseases (SINAN) through the Department of Epidemiological Surveillance of the Health Department of the Municipality of Caxias, bank information DATASUS and Center for Zoonoses (CCZ) in Caxias, Maranhão, since January 2003 to May 2010 with a convenience sample. Results: During the period in question were observed 333 human cases of VL, while 10 cases resulted in death (mortality 3.0%). The annual distribution of cases was 77 cases in 2003, 37 in 2004, 54 in 2005, 21 in 2006, 47 in 2007, 42 in 2008, 34 in 2009 and 21 cases registered till May 2010. Most cases occurred in children aged between 1 and 4 years, 143 cases (42.9%), but other age groups were also affected. Prevalent among males with 56.7% of cases and observed that the LV in the city of Caxias is predominantly urban. Most patients had parasitological confirmation, the clinical and epidemiological criterion alone was not relevant. The pentavalent antimony was the drug of first choice in the first treatment in 93.7% of cases. In canine serological surveys conducted in the period 2005 to 2009, 47.3% of canine serum collected was positive. Conclusions: There were no deaths in the last three years, meaning low severity of disease in the city, suggesting that the symptoms and factors associated with unfavorable clinical course of patients with VL in Caxias been prevented and / or addressed at an early stage, reflecting the good quality assistance in the municipality of Caxias. It is noteworthy that still makes it necessary to raise awareness of the government, especially in regard to controlling the transmission.

- EPIDEMIOLOGY [504] P 734 - GEOGRAPHIC DISTRIBUTION OF CUTANEOUS LEISHMANIASIS (CL) IN THE PERIOD FROM 2008 TO 2012 IN THE CITY OF BARREIRINHAS, MARANHÃO, BRAZIL. FILHO, A.A.P.1; FONTELES, R.S.2; BANDEIRA, M.C.A.2; MORAES, J.L.P.2; SILVA, S.O.1; REBÊLO, J.M.M.2; MELO, M.N.1 1.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 2.UNIVERSIDADE FEDERAL DO MARANHÃO, SÃO LUÍS, MA, BRAZIL.

Keyword:cutaneous leishmaniasis; geographic distribution; barreirinhas

Abstract: The CL is caused by protozoa of the genus Leishmania, transmitted by sand flies diptera family Psychodidae. In Brazil, the cases are related mainly to the environmental degradation (such as deforestation), lack of investment in education and health infrastructure. Inserted in this context, Barreirinhas, located in Maranhão, has the most of the cases of CL in the state, about 40%. It is related with the tourism, the main economic activity of the region, developed in a predatory way and without proper planning. In addition, the massive deforestation in rural areas, due to the migration of families to these areas, contributes to the increase of cases. Therefore this study aimed to analyze eco-epidemiological aspects of LTA in the villages of Barreirinhas. In 2008, 75 cases were registered with distribution in 39 locations in and around Barreirinhas headquarters, where 08 of the 75 cases occurred in urban areas and the rest in rural areas with the highest concentration in the villages Armazém (04), Baixa D’água (04), Jabuti (06) and Palmeira dos Eduardos (10), this year enrolls approximately 38.26% of the cases. In 2009, the number of positive cases detected in 20 locations was 35, involving new locations such as villages: Pedras, Borges, the district arnaubal (urban area). In 2010, the number of cases suffer a new jump to 61 records of positive LTA involving 35 locations, with the highest concentration in the villages, Piquizeiro (05), Baixa D’Água II ( 06) and Guaribinha (06). In 2011, 48 sites were registered with CL positive cases totaling the highest number since 2008, 81 cases.

Between the new affected villages worth mentioning Engenho, Gambá and São Domingos. In the year 2012, the number of cases was 77 distributed in 45 locations, with the highest

concentration in the villages: Fome (10), Barrras (06) and Mangas (05). The city area Barreirinhas is advancing, affecting population growth in the areas of their natural surroundings.

In addition, the developmet of agricultural activities in rural areas contributes to degradation of riparian areas and to the promotion of human contact with the vector. Thus, studies about the distribution and dynamics of the vector of this disease endemic in the city and its environments are required.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [505] P 735 - STUDY OF THE PHLEBOTOMINE FAUNA IN FORMIGA, MINAS GERAIS STATE, BRAZIL MORAIS, M.H.G.1; MENEZES, J.A.2; SOUSA, A.M.1; DA FONSECA, A.C.3; FILHO, V.P.3; DILERMANDO, J.A.2; DE SOUZA, C.M.2

1.FUNDAÇÃO EDUCACIONAL DE DIVINÓPOLIS, UNIVERSIDADE DO ESTADO DE MINAS GERAIS, DIVINÓPOLIS, MG, BRAZIL; 2.CENTRO DE PESQUISAS RENÉ RACHOU, FUNDAÇÃO OSWALDO CRUZ, BELO HORIZONTE, MG, BRAZIL; 3.CENTRO UNIVERSITÁRIO DE FORMIGA, FORMIGA, MG, BRAZIL.

Keyword:phlebotomine; ecoepidemiology; formiga (minas gerais state)

Abstract: Introduction: Study of the phlebotomine fauna in foci of recent transmission of leishmaniasis is essential for epidemiological surveillance of the disease, therefore will clarify about the main existing species and their distribution. In Formiga, Minas Gerais, the first case detected of Visceral Leishmaniasis (VL) occurred between 2011 and 2012, however, the micro-region where the municipality is located is endemic for American Cutaneous Leishmaniasis (ACL).

Thus, the aim of the present study was to evaluated the local phlebotomine fauna and identify potential areas of the transmission risk. Methods: Monthly, systematic collection of sandflies are being held in the peridomicile of 24 domiciles, using HP light traps for four days consecutively, from 5 p.m. to 7 a.m. The households were selected by convenience in accordance with information from the Municipal Health Department about cases of canine and human Leishmaniasis and the presence of propitious environmental to phlebotomines development. The insects captured are being mounted between slide and coverslip for taxonomic identification using the key of Young and Duncan (1994). Partial Results: From May to October of 2012 were captured 82 phlebotomines of 6 species: 32 Lutzomyia longipalpis (39,02%), 23 Lutzomyia cortelezzi (28,04%), 19 Lutzomyia whitmani (23,17%), 6 Lutzomyia lenti (7,31%), 1 Lutzomyia monticola (1,21%) and 1 Lutzomyia termitophila (1,21%). The first three species were the most abundant during the capture. The highest density of phlebotomines was observed in September, probably due to increased precipitation recorded in that period. The occurrence of VL cases and the gathering of L. longipalpis indicate that, probably, this species is associated with the transmission of the disease in the city, but more studies are required. Furthermore, the incidence of L. whitmani (ACL vector) confirms the necessity of vigilance in the region. Considerations: The encounter of species demonstrably vectors of VL and ACL reinforces the necessity of entomological surveillance in order to prevent the increase of VL cases and the onset of ACL cases in the city. These results associated with the study of canine infection that our group is conducting in the municipality, would give subsidies to map the areas at risk and direct the necessary actions to the control of Leishmaniasis in the region.

- EPIDEMIOLOGY [509] P 736 - AUTOCHTHONOUS CASES OF LEISHMANIASIS IN DOGS FROM FLORIANÓPOLIS, SANTA CATARINA, BRAZIL PACHECO, A.D.1; TOMOKANE, T.Y.2; LIMA, V.M.F.1; LAURENTI, M.D.2; MARCONDES, M.1 1.SCHOOL OF VETERINARY MEDICINE, SÃO PAULO STATE UNIVERSITY, ARAÇATUBA, SP, BRAZIL; 2.SÃO PAULO UNIVERSITY, SÃO PAULO, SP, BRAZIL.

Keyword:zoonotic visceral leishmaniasis; serology; real-time pcr

Abstract: A total of 491 healthy dogs from the city of Florianópolis, Santa Catarina state, a non endemic area for zoonotic visceral leishmaniasis until 2012, when three cases were first reported, were serologically evaluated by enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFAT) for leishmaniasis. ELISA cut-off value (mean value plus 3 SD) was determined by analysis of serum samples from 30 healthy dogs from a non-endemic area for leishmaniasis. IFAT cut-off value was 1:40. The occurrence of cross-reaction with Ehrlichia canis and Dirofilaria immitis was discharged testing serum samples by a commercial kit (SNAP 4DX, IDEXX Laboratories). Since Santa Catarina state is a non endemic area for Chagas’ disease, free of vector and with no reported case to date, cross-reaction with T. cruzi was also rulled out. Whole blood from dogs with positive results in any of the techniques was also submitted to real-time PCR (qPCR) for Leishmania sp. DNA detection. The population was composed by 337 (68.6%) females and 151 (31.4%) males from 52 (61%) of the 85 neighbourhoods of Florianópolis. Ages ranged from six months to 12 years, with an average of three years. None of the evaluated dogs had visited other counties and all of them were born in Florianópolis. The total seroprevalence of leishmaniasis was 5.3% (25/491); 0.4% (2/491) by ELISA and 4.9% (24/491) by IFAT. In just one dog seroreactivity was observed simultaneously in both serological methods. That was also the only dog in which parasite DNA was amplified in whole blood by qPCR. Seroreactivity occurred in dogs from 14.11% (12/85) of the neighbourhoods of the city. The results of this study confirm the occurrence of autochthonous cases of leishmaniasis in dogs in Florianópolis, and highlight the necessity of an accurate investigation in order to implement control strategies since they can be cases of visceral or cutaneous leishmaniasis.

- EPIDEMIOLOGY [516] P 737 - BURDEN OF INFECTION IN COMMUNITIES EXPERIENCING DOMESTIC TRANSMISSION OF DERMAL LEISHMANIASIS IN RISARALDA, COLOMBIA RUBIANO PEREA, L.C.; COSSIO, A.; JOJOA, J.; ROSALES, M.; BLANCO, V.M.; GOMEZ, M.A.; GORE SARAVIA, N. CIDEIM, CALI, COLOMBIA.

Keyword:burden; domestic transmission; prevalence

Abstract: Introduction: Domestic transmission and urbanization of Leishmania Viannia species have become frequent in Colombia and neighboring countries. Contributing factors include population displacement, deforestation and adaptation of sand flies to peridomestic settings.

Domestic transmission impacts the most vulnerable population groups: women, children and the elderly. This study sought to determine the age distribution and burden of infection among inhabitants of communities experiencing domestic transmission of cutaneous leishamniasis (CL) Methodology: Age-specific prevalence of symptomatic and subclinical infection was determined in the communities, Itaurí and La Cabaña in Risaralda, Colombia, where sand flies were present within households and children presented healed or active cutaneous leishmaniasis. Prevalence of infection was defined based on clinical and immunological evidence (skin test) and parasitologically using PCR/Southern blot of kDNA and qRTPCR of 7 SLRNA in blood and mucosal tissues (conjunctiva, tonsils, nasal mucosa).

Results: The communities presented a pyramidal population profile with 46% 20 years of age.

Among the 180 inhabitants, 124(69%) participated in the study. Overall prevalence of

symptomatic infection was 70% (n=87, 68% historic; 2% active), asymptomatic infection 8% (n=10) and uninfected 22%, (n=27). Prevalence of symptomatic infection increased from 48% in the first decade of life to 78% by the second decade. Subclinical infection ranged from 3% of 010 year olds to 19% of residents > 50 years of age. Leishmania DNA was detected in blood or mucosal tissues of 50% of participants with history of CL or positive skin test between 11 and 50 years of age (n=85) and 10% of children 10 years of age (n=20). Parasite DNA was detected in blood of 37% and mucosal tissue of 21% of participants with symptomatic or subclinical infection.

Conclusions: High prevalence of infection was documented in settings of domestic transmission based on clinical, immunological and molecular evidence. Considering that one-time sampling provides a minimum estimate, amplification of Leishmania DNA from at least 50% of residents with prior CL or subclinical infection confirmed persistent Leishmania (Viannia) infection in a large proportion of the community. The burden of infection in these and similar settings presents previously unrecognized challenges to control: incident disease will involve children and reactivation of prior infection.

- RESERVOIRS [519] P 738 - LEISHMANIA (VIANNIA) BRAZILIENSIS IN EQUIDS FROM AREAS ENDEMIC FOR AMERICAN CUTANEOUS LEISHMANIASIS IN SOUTHERN BRAZIL: SEROLOGICAL AND MOLECULAR STUDY THOMAZ SOCCOL, V.T.1; TRUPPEL, J.H.2; OTOMURA, F.3; TEODORO, U.3; MASSAFERA, R.4; DALAGRANA, L.5; CATARINO, C.M.6; COSTA-RIBEIRO, M.C.V.6; COSTA FERREIRA, M.E.7 1.UNIVERSIDADE POSITIVO, CURITIBA, PR, BRAZIL; 2.CENTER FOR ZOONOSIS CONTROL, DEPARTMENT OF HEALTH SURVEILLANCE, MUNICIPAL HEALTH DEPARTMENT, ARAUCÁR, ARAUCÁRIA, PR, BRAZIL; 3.POSTGRADUATE PROGRAM IN HEALTH SCIENCES, MARINGÁ STATE UNIVERSITY, MARINGA, PR, BRAZIL; 4.SURVEILLANCE SERVICE OF HEALTH, MINISTRY OF HEALTH, BRAZIL, JACAREZINHO, PR, BRAZIL; 5.UNIVERSIDADE FEDERL DO PARANÁ, CURITIBA, PR, BRAZIL; 6.UNIVERSIDADE FEDERAL DO PARANÁ, CURITIBA, PR, BRAZIL; 7.GEOGRAPHY DEPARTMENT, MARINGÁ STATE UNIVERSITY (UEM), MARINGA, PR, BRAZIL.

Keyword:leishmania braziliensis; molecular epidemiology; reservoir

Abstract: Cutaneous leishmaniasis (CL) is an endemic disease worldwide, an important zoonosis that presents constant, wide geographic expansion due to human-induced deforestation conducted to increase agricultural and pastoral areas. These environmental changes are creating a favorable scenario for the emergence or reemergence of CL and the possibility of new epidemiological patterns of the disease. In this study we detected the infection of equids with L. (Viannia) braziliensis from southern Brazil, in CL endemic areas. Antibodies to the parasite were assayed using the Enzyme Linked Immunosorbent Assay. Blood samples were collected and tested by polymerase chain reaction (PCR), and the positive products were sequenced. The results showed that 11.0% (25/227) equids were seropositive for L. (V.) braziliensis and 16.3% (37/227) were PCR positive. anti-L. (V.) braziliensis antibodies were detected in 20 horses, 3 donkeys and 2 mules and the parasite DNA was detected in 30 horses, 5 donkeys and 2 mules. Sequencing the amplified DNA revealed 100% similarity with sequences for Viannia complex, corroborating the results of PCR for L. (V.) braziliensis. Our results show that equids are infected with L. (V.) braziliensis, therefore they could be food sources for phlebotomines in the peridomiciliary environment and consequently play a role in the CL cycle.

- EPIDEMIOLOGY [523] P 739 - MORTALITY RELATED TO VISCERAL LEISHMANIASIS AND HIV/AIDS COINFECTION IN BRAZIL, 2000-2010 LIMA, M.S.; MARTINS-MELO, F.R.; ALENCAR, C.H.M.; HEUKELBACH, J.; RAMOS JÚNIOR, A.N. UNIVERSIDADE FEDERAL DO CEARÁ, FORTALEZA, CE, BRAZIL.

Keyword:visceral leishmaniasis; hiv/aids; brazil

Abstract: Visceral leishmaniasis (VL) is considered an opportunistic disease in people with HIV/AIDS. Co-infection may reduce the survival of patients significantly, as VL may modify the progression of HIV/AIDS. On the other hand, HIV-induced immunosuppression facilitates progression of VL. This study describes the epidemiological aspects of deaths related to VL and HIV/AIDS co-infection in Brazil, from 2000 to 2010. We analyzed mortality data obtained from the Mortality Information System (Sistema de Informação sobre Mortalidade - SIM /MS /DATASUS) and included deaths that occurred in which VL (ICD-10: B55.0) and HIV/AIDS (ICD-10: B20-B24) were mentioned in the same death certificate (Declaração de óbito - DO), either as underlying or associated causes of death (multiple causes of death). During this period, the association of VL with HIV/AIDS was mentioned in 230 deaths (7.6% of deaths by VL).

AIDS was an underlying cause in 60.9% (140), while VL was the underlying cause in 38.3% (88) of deaths. VL and AIDS were presented as associated causes in another 2 deaths.

Predominating characteristics were: male gender (78.7%), brown race/color (56.1%), singles (64.3%), age 30-39 years (41.3%), resident in the Northeast region (43.9%) and from all states in Brazil the state of Minas Gerais has the largest proportion of cases (21.7%). Despite the majority residing in rural municipalities (62.2%), the place of occurrence of deaths was mostly in hospitals (97.3%) of the Brazilian state capitals (73%). The systematic description of the epidemiological characteristics and magnitude of mortality related with VL and HIV/AIDS coinfection reflects the need to adopt special strategies to monitor this group. The ruralization of HIV and urbanization of VL in Brazil cause an increasing geographical overlap, representing an important public health problem. Control measures will require comprehensive and reliable information, and an integration of the surveillance of VL and AIDS is needed, as well as enhancement of surveillance of VL and HIV/AIDS co-infection. We recommend the systematic use of HIV testing in patients with VL.

- EPIDEMIOLOGY [526] P 740 - GEOGRAPHIC DISTRIBUITION OF HUMAN AND CANINE VISCERAL LEISHMANIASIS CASES IN THE CITY OF ARACAJU, SERGIPE, BRAZIL TUNON, G.I.; DE MOURA, T.R.; RANGEL, M.U.; SANTANA, A.P.; DE MELO, E.V.; ARAGAO, M.T.; DOS SANTOS, P.L.; DA COSTA, R.V.; LEITE, M.O.; LIMA, S.S.; ALMEIDA, J.A.; ALMEIDA, R.P. UNIVERSIDADE FEDERAL DE SERGIPE, ARACAJU, SE, BRAZIL.

Keyword:geoprocessing; leishmaniasis; spatial analysis

Abstract: The change in the epidemiological profile of visceral leishmaniasis from wild / rural to urban environment is a fact that has been occurring in Brazil’s Northeast region. The use of geoprocessing in conjunction with satellite images are extremely useful tools that can be used in epidemiological studies of endemic diseases transmitted by vectors. Considering that there are

few Brazilian studies of canine visceral leishmaniasis based on geoprocessing techniques, we used this approach to determine the spatial distribution patterns of canine and human cases in the city of Aracaju between 2005 and 2012. The aim was to compare the presence of the disease by neighborhood and the distribuition of nitric oxide resistant Leishmania isolates. The study area comprises the area of the county of Aracaju, which is divided in 37 neighborhoods and is located in the coastal area of the state of Sergipe. We georeferenced 2,060 positive dogs and 110 human cases from 2005 to 2012 from data collected from the Zoonosis Control Center and the UFS University Hospital. The cases were spatialized according to geographical location of the address of each reported case. These data were tabulated and exported to Spring 5.1.5 software to generate thematic maps showing the distribution of leishmaniasis cases by neighborhoods of the city of Aracaju. We observed that the human cases of the city happened in areas with relatively high density of leishmaniasis positive dogs, confirming that the occurrence of canine cases precedes the occurrence of human cases. Most human (18) and canine (676) cases occurred in the southern city neighborhoods of Zona de ExpansĂŁo and Mosqueiro. Comparing the spatial distribution, we noticed that the neighborhoods most affected were those in peripheral areas with lower population densities, recent occupation, poor urban infrastructure and low income. In other neighborhoods close to downtown area (Olaria, Santos Dumont, America and Jardim CentenĂĄrio), also showed significant cases of human and canine disease. In three neighborhoods (Santos Dumont, America and Dezoito do Forte) where isolated nitric oxide resistant Leishmania from both human and dogs, fact that may be related to a more aggressive form of the disease. The use of geoprocessing techniques along with relevant information of the study area allowed visualization of the geographical distribution of this disease in Aracaju and further facilitate the implementation of appropriate control measures.

- CONTROL PROGRAMS [528] P 741 - PERFORMANCE OF KALA-AZAR SURVEILLANCE IN GAFARGAON SUBDISTRICT OF MYMENSINGH, BANGLADESH RAHMAN, K.M.1; MONDAL, D.2; LUBY, S.P.3; SLEIGH, A.C.1

1.AUSTRALIAN NATIONAL UNIVERSITY, CANBERRA, AUSTRALIA; 2.INTERNATIONAL CENTRE FOR DIARRHOEAL DISEASE RESEARCH, BANGLADESH, DHAKA, BANGLADESH; 3.STANFORD UNIVERSITY, STANFORD, UNITED STATES.

Keyword:kala-azar; surveillance; performance

Abstract: Background: The goal of South Asia kala-azar elimination is to lower annual incidence to less than 1 per 10,000 at the district or sub-district level. In Bangladesh, government passive surveillance generated by monthly sub-district reports is being used to monitor progress with the elimination program. We studied performance of the kala azar passive surveillance system in a highly endemic sub-district of Bangladesh. Methods: We conducted a cross-sectional survey from September 2011 to June 2012 in randomly sampled villages of Gafargaon upazila (subdistrict) of Mymensingh district. We screened for kala-azar cases since January 2010, defining cases as those diagnosed by a qualified health care provider with a laboratory test for kala-azar positive. To cross-check with passive surveillance, those identified by our active screening were searched for in the hospital register of the Gafargaon upazila (sub-district) Health Complex (UHC). Results: Of the 67 kala-azar cases detected by screening, we dropped 6 cases from our analysis because they went to a government health facility outside Gafargaon or joined a clinical trial in Gafargaon. Among the remaining 61 cases, 29 (48%) were reported in the (passive surveillance) Gafargaon UHC patient register and eventually contributed to national kala-azar tallies. This proportion increased from 42% in 2010 to 57% in 2011. Conclusion: Around half of the kala-azar cases occurring in a highly endemic sub-district of Bangladesh were not reported through the national passive surveillance system. Larger samples from regions of different levels of endemicity would help confirm these findings. This will eventually contribute to the population based estimation of kala-azar incidence of the country.

- RESERVOIRS [531] P 742 - THE ROLE OF THE DOG IN AN ENDEMIC AREA OF CUTANEOUS LEISHMANIASIS MARQUEZ, E.S.1; NAVARRO, I.T.2; NABUT, L.B.2; CASTRO, E.A.3; COUBEU, S.B.3; RIBEIRO, M.3; FORDELLONE, M.R.C.4; THOMAZ-SOCCOL, V.3 1.UNIVERSIDADE ESTADUL DO NORTE DO PARANÁ, BANDEIRANTES, PR, BRAZIL; 2.UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL; 3.UNIVERSIDADE FEDERAL DO PARANÁ, CURITIBA, PR, BRAZIL; 4.UNIVERSIDADE ESTADUAL DO NORTE DO PARANÁ, BANDEIRANTES, PR, BRAZIL.

Keyword:cutaneous leishmaniasis; dogs; reservoir

Abstract: The role of the dog as a reservoir of L. infantum has been confirmed, but the same does necessarily not apply to other species, such as L. (V.) braziliensis, which inhabits the largest geographical area in Brazil. Using parasitological, serological and molecular methods, we investigated the presence of Leishmania in different tissues (skin lesions, intact skin, spleen, liver, lymph nodes, bone marrow and peripheral blood) of naturally infected dogs to elucidate the role of the dog in maintaining the cycle of the parasite. To achieve this, 48 (80%) samples from naturally infected dogs with lesions suggestive of American cutaneous leishmaniasis were evaluated. Twenty-eight dogs (58.3%) were positive for serology and isolation of the parasite in vitro. Eight dogs with positive parasitological diagnosis were euthanized in accordance with the regulations of the Brazilian Health Surveillance Agency. During necropsy, blood and several other tissues were collected for parasite research by tissue culture and PCR. The parasite was isolated in culture medium of skin ulcer samples from all the dogs euthanized. Parasite identification by RAPD-PCR determined that the species was L. (V.) braziliensis. Parasite DNA was screened for in tissue lesion samples, using the PCR technique, and seven (87.5%) dogs presented amplified fragments of 70 base pairs. Whole blood of four out of eight dogs (50%) was positive for parasite DNA. The amplified DNA fragments were submitted to sequencing, which presented 100% similarity with the Viannia complex. Positivity was determined in more than one tissue fragment from each dog. The percentages for the different organs investigated were: 100% parasite isolation in skin lesions, 87.5% in bone marrow samples, 62.5% in liver samples, 50% in lymph nodes and blood samples and 37.5% in spleen and intact skin samples.

The best explanation for the presence of the parasite in different organs is that following inoculation of Leishmania by the vector, the parasite is preferentially located in the skin, with hematogenous dissemination occurring after an unknown period, such that the concentration of circulating parasites is lower and, consequently, is also lower in other organs than at the skin lesion site. Since the parasite was identified in blood and tissue and the dogs’ period of survival was greater than 12 years, domestic canines may well be good reservoirs for L. (V.) braziliensis.

- EPIDEMIOLOGY [533] P 743 - MOBILITY AND SPATIAL DISTRIBUTION OF CUTANEOUS LEISHMANIASIS IN THE LEGAL AMAZONIA, 2007-2009 VALDES, A.C.; OLIVEIRA, R.M.; SABROZA, P.C. ENSP/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:cutaneous leishmaniasis; ecosistema amazónico; spatial distribution

Abstract: Cutaneous Leishmaniasis (CL) is an endemic disease in Brazil. Its transmission depends on specific features, such as vectors strictly limited to certain environmental factors, diversity of agents and reservoirs. Displays transmission patterns urban and rural, related to the type of work shifts to endemic areas and housing type of infected individuals, which makes it necessary to

know the territory and the flow of people in areas where the disease is inserted for surveillance actions . The aim of this study was to correlate the spatial mobility of people and the spatialtemporal distribution of Cutaneous Leishmaniasis, in Amazonia, in the period from 2007 to 2009, considering the flows from the territory of the municipality of residence to the municipality suspected infection. Descriptive epidemiological study focused on understanding the relationship between mobility and spatial distribution of cases of CL in the Legal Amazonia, identifying areas of epidemiological relevance, according to the municipality of infection and considering flows from the territory of the municipality of residence to the municipality suspected infection. Was identified 09 circuits of production of CL, consisting of 15.01% of the municipalities in the region, with 15.182 cases (39.71%), detection average 46.14 cases per 100.000 inhabitants. and average density 4.40 cases/km2, coinciding with the regions of the circuits when analyzed by municipality of residence, but with reduced quantity of municipalities involved. Regarding flows, 5.891 cases (11.69%) present municipality of residence different from the municipality of infection and identified numerous streams with reduced average cases.

These flows usually occurred between neighboring municipalities. Regarding poles difference was observed between the areas considered most important for the disease, according to different criteria. Data analysis using municipality of infection and flows allows people realize the difference the poles of epidemiological relevance when used different criteria. Currently, we use only the municipality of residence. This combined with the other criteria, enables the improvement of surveillance, identification of transmission patterns, new areas of epidemiological relevance, especially those related to work processes, enabling improved disease control.

- EPIDEMIOLOGY [538] P 744 - CHARACTERIZATION OF PARAGOMINAS/AP AS A POLE OF EPIDEMIOLOGICAL RELEVANCE FOR LEISHMANIASIS IN LEGAL AMAZONIA, 2007 TO 2009.

VALDES, A.C.; OLIVEIRA, R.M.; SABROZA, P.C. ENSP/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:cutaneous leishmaniasis; amazon ecosystem; descriptive epidemiology

Abstract: Introduction: In Brazil, Cutaneous Leishmaniasis is an endemic disease. Specific characteristics are related to its transmission, as vectors strictly limited to certain environmental factors and diversity of agents and reservoirs. Its distribution is also related to the flow of people in the territory, mainly related to shifts in the work process. How important area of concern related to the flow of cases and the distribution of Cutaneous Leishmaniasis was identified municipality Paragominas/AP. Objective: To characterize the city of Paragominas a pole of epidemiological relevance for Cutaneous Leishmaniasis in the Legal Amazonia, according to density of cases within a circuit and flow of cases within the period of 2007/2009. Material and Methods: descriptive epidemiological study focused on identification and characterization of a pole of epidemiological relevance for LT. The identification was made by the high density of cases within a circuit, high number of flows of people living in the city by scrolling up and infecting others with the disease, and residents of other municipalities who became infected in polo study, period 2007/2009. And characterization by the analysis of environmental and economic indicators. Results: Paragominas belongs to the Circuit Tucuruí. The municipality has a geographic position that gives advantages to the flow of production through the Belém-Brasília highway (BR-010). Receives significant amount of people seeking employment opportunities, driven by the mining company VALE, engaged in bauxite mining. It is also a major timber poles Legal Amzonia. There was population growth from 2000-2010 in relation to its upper circuit, but with poor urban structure. About Cutaneous Leishmaniasis, during the period, 119 cases were infected in Paragominas and lived in other cities and other 100 cases were infected in other

municipalities and resided in polo. Conclusion: The environmental indicators indicate an economic and population growth of the city, however, urban development has not kept pace.

Regarding the flow of people who become infected with Cutaneous Leishmaniasis, these individuals may be being drawn to the region in search of new job opportunities. At the same time, the precarious structure for fixing within this population generates large output stream to nearby towns in search of other opportunities.

- EPIDEMIOLOGY [539] P 745 - EXPANSION OF VISCERAL LEISHMANIASIS IN TRÊS LAGOAS - MS ZUQUE, M.A.S.1; ZUQUE, F.T.S.1; ZUQUE, F.R.S.2; LUCHEIS, S.B.3; EMPKE, A.L.T.4; GIMENEZ, D.S.B.4; PETRONI, T.F.1 1.FACULDADES INTEGRADAS DE TRÊS LAGOAS - AEMS, TRÊS LAGOAS, MS, BRAZIL; 2.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL - UFMS / CPCX, COXIM, MS, BRAZIL; 3.AGÊNCIA PAULISTA DE TECNOLOGIA DOS AGRONEGÓCIOS (APTA/SAA), PÓLO REGIONAL CENTRO-OESTE, BAURU, SP, BRAZIL; 4.CENTRO DE CONTROLE DE ZOONOSES DE TRÊS LAGOAS, TRÊS LAGOAS, MS, BRAZIL.

Keyword:urbanization; visceral leishmaniasis; endemicity

Abstract: Introduction: There are numerous difficulties in adopting effective measures in urban centers related to the control of Visceral Leishmaniasis (VL), the proposed measures to control are: application of insecticides, diagnosis and appropriate treatment of reported cases, control of the canine reservoir and euthanasia of seropositive dogs to control the strength of canine infection.

Objectives: To describe the behavior of the LV at Três Lagoas / MS (Brazil), through control actions recommended by the Ministry of Health. Methodology: Was analyzed data from sectors that: endemic, epidemiological surveillance and the Center for Zoonosis Control ( CCZ) for the period 2007 to 2011. Results: The first cases of Canine Visceral Leishmaniasis (CVL) in the urban area of Tres Lagoas occurred in 1999 and the diagnosis of Human Visceral Leishmaniasis (LVH) in 2000. Was recorded 266 cases of LVH in the period from 2000 to 2006, and the lethality rate ranged from 6% to 50%. There was a decrease in cases from 2007 to 2012, with a record of 115 cases of LVH and lethality rate ranging from 4% to 25%. Were collected for euthanasia the symptomatic dogs or reactives for LV, with 3302 dogs in 2007, 3551 in 2008, 3681 in 2009, 3440 in 2010, 3774 in 2011. The positivity rate in canine surveys of areas of intense transmission ranged from 29% in 2007 to 8% in 2011. Discussion: Was considered factors responsible for the emergence and maintenance of LV cases in the city: a geographic location that allows communication with endemic centers of the disease, environmental change caused by deforestation that changes in the ecosystem of the city; growth and spread of L. longipalpis currently detected throughout the urban area. The dog seems to be the facilitating factor of urbanization and the LV was observed by the high replacement canine population.

Since 2007 the city has stepped up measures to control LV and observed a decrease in LVC in areas of intense transmission, sprayed annually and dogs were reactive collected for euthanasia, but there was an increase in LVC in other areas of the city fact observed by the gathering of symptomatic dogs by CCZ. Conclusion: The results suggest the expansion, urbanization and endemicity of the disease in the city with the appearance of cases in each year of the period studied by varying its intensity.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [551] P 746 - DISTRIBUTION OF LUTZOMYIA LONGIPALPIS CHEMOTYPE POPULATIONS IN SÃO PAULO STATE, BRAZIL. CASANOVA, C.1; HAMILTON, J.G.C.2; COLLA-JACQUES, F.E.3; BRAZIL, R.P.4; SHAW, J.J.5

1.SUPERINTÊNDENCIA DE CONTROLE DE ENDEMIAS, SECRETARIA DA SAÚDE DO ESTADO DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 2.CENTER FOR APPLIED ENTOMOLOGY AND PARASITOLOGY, UNIVERSITY OF KEELE, KEELE, UNITED KINGDOM; 3.INSTITUTO DE BIOLOGIA, PROGRAMA DE PÓS-GRADUAÇÃO EM PARASITOLOGIA, UNIVERSIDADE DE

CAMPINAS, CAMPINAS, SP, BRAZIL; 4.INSTITUTO OSWALDO CRUZ, FUNDAÇÃO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL; 5.INSTITUTO DE CIÊNCIAS BIOMÉDICAS, UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL.

Keyword:lutzomyia longipalpis; sex pheromone; são paulo state

Abstract: Information on the geographical distribution, dispersal mechanisms and route tendencies of insect-borne diseases is epidemiologically important. It can help to identify ongoing transmission areas, new risk areas and guide surveillance and control activities. Lutzomyia longipalpis (Lutz & Neiva), the principal vector of Leishmania infantum chagasi (Cunha & Chagas) in the Americas, is a species complex that can be separated according to the type of pheromone. It is still unclear how many members there are in this complex, how they are related and if some are more important vectors than others. Recently, we identified two populations in São Paulo State: a 9-methylgermacrene-B pheromone producing population found in the municipalities of Araçatuba, Dracena, Promissão, and Bauru (located in the western region of the state); and a Cembrene-1 pheromone producing population in the municipality of Espírito Santo do Pinhal, located in the north-eastern region of the state. Both chemotype populations were found in peri-urban areas of São Pedro, located in the central region of the state. Human cases have only been recorded in municipalities located in the western region of the state where L. longipalpis 9-methylgermacrene-B population is collected in large numbers and where the majority of the infested municipalities are located. The remarkable differences between the epidemiological situation and the population size of L. longipalpis in these two regions suggest that 9-methylgermacrene-B may have increased vectorial capacities compared to cembrene-1 populations. The number of municipalities harbouring L. longipalpis and the numbers of canine and human AVL cases are all increasing every year in São Paulo state. The present study updates the geographical and progressional distribution of canine and human AVL and its vector. Also the chemotype of L. longipalpis populations from Presidente Prudente, Adamantina, Osvaldo Cruz, Campinas, Itupeva, Sorocaba, Salto, Socorro, Marilia (and others) is presented, and the possible dispersion routes of the different populations are discussed.

- EPIDEMIOLOGY [552] P 747 - LEISHMANIA SPECIES IDENTIFICATION AND LEISHMANIAVIRUS DETECTION ON CLINICAL SAMPLES FROM CUTANEOUS AND MUCOCUTANEOUS LEISHMANIASIS PATIENTS IN RONDÔNIA, WESTERN AMAZONIAN REGION. CANTANHEDE, L.M.1; SILVA JÚNIOR, C.F.2; ITO, M.M.2; CUSTÓDIO, M.G.F.1; ALVES, P.H.1; NICOLETE, R.1; SALCEDO, J.M.V.1; GARRIDO, L.M.3; PESCARINI, J.M.3; KRIEGER, H.3; FERREIRA, R.G.M.1 1.FIOCRUZ RONDÔNIA, PORTO VELHO, RO, BRAZIL; 2.UNIVERSIDADE FEDERAL DE RONDÔNIA, PORTO VELHO, RO, BRAZIL; 3.UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL.

Keyword:leishmania species; leishmaniavirus; cutaneous and mucocutaneous leishmaniasis

Abstract: The American tegumentary leishmaniasis (ATL) is endemic throughout the Amazon region and the state of Rondonia is the third Brazilian state in number of reported cases, with an average of 1000 new cases per year. In Amazonia, seven different species of Leishmania have been described. Clinical manifestation may range from a cutaneous form to the destruction of the mucosal tissues. The mechanisms that lead to disease progression to the mucosal form in about 10% of patients treated or under treatment remain unclear and might be related with parasite and host genetic factors, immune response and more recently the presence a virus infecting the pathogen, named Leishmaniavirus (LRV). The objective of the present study was to identify the Leishmania species causing ACL in Rondonia, Brazilian western Amazonian region, as well as

verify if the LRV could be detected among studied patients and its possible association with the presented clinical form. Cytology brush cotton swabs were collected from lesions of 137 patients with clinical suspicion of leishmaniasis and stored immediately in 1 ml of RNALater®.

All samples were subjected to PCR amplification and species identification by RFLP with primers to the internal transcribed spacer 1 (ITS1), to the heat shock protein (HSP70) as well as kinetoplastide DNA (kDNA) – only for leishmaniasis diagnosis. 121 patients presented positive results for at least one of the molecular tests applied. The most frequent species found using HSP70 was Leishmania braziliensis 73 (60.3%), followed by Leishmnia guyanensis 33 (27.3%), Leishmania shawi 2 (1.7%), Leishmania amazonensis 1 (0.8%) and mixed infection was L. guyanensis and L. braziliensis 3 (2.5%), L. guyanensis and L. amazonensis 1 (0.8%). Species from 8 (4.2%) patients were not identified. The cutaneous form was found on 78 patients, mucous form on 38, both forms on 4, and 1 patient presented the diffuse form. For LRV detection, RNA was extracted from the positive samples and cDNA synthesized for PCR amplification using primers specific for the virus genome. The virus was detected in 57 patients (47.1% - 95%CI 38.1%...56.0%). A border line association (p = 0.074, Fisher’s exact test) was found between the presence of LRV and the form of the lesion (excluding the diffuse and mixed forms patients). Overall, the risk of been positive for LRV among cutaneous form patients was 0.772 (95%CI 0,591…1.007) and among mucous for patients was 1,702 (95%CI 0.992…2.915).

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [558] P 748 - LARVAL BREEDING SITES OF LUTZOMYIA LONGIPALPIS (DIPTERA: PSYCHODIDAE) IN VISCERAL LEISHMANIASIS ENDEMIC URBAN AREAS IN SOUTHEASTERN BRAZIL CASANOVA, C.1; ANDRIGUETTI, M.T.M.1; SAMPAIO, S.M.P.1; MACORIS, M.L.G.1; COLLAJACQUES, F.E.2; PRADO, A.P.3 1.SUPERINTÊNDENCIA DE CONTROLE DE ENDEMIAS, SECRETARIA DA SAÚDE DO ESTADO DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 2.INSTITUTO DE BIOLOGIA, PROGRAMA DE PÓS-GRADUAÇÃO EM PARASITOLOGIA, UNIVERSIDADE ESTADUAL DE CAMPINAS, CAMPINAS, SP, BRAZIL; 3.INSTITUTO DE BIOLOGIA, UNIVERSIDADE ESTADUAL DE CAMPINAS, CAMPINAS, SP, BRAZIL.

Keyword:breeding sites; lutzomyia longipalpis; são paulo state

Abstract: The scarcity of information on the immature stages of sand flies and their preferred breeding sites has resulted in the focus of vectorial control on the adult stage using residual insecticide house-spraying. This strategy, along with the treatment of human cases and the euthanasia of infected dogs, has proven inefficient and visceral leishmaniasis continues to expand in Brazil.

Identifying the breeding sites of sand flies is essential to the understanding of the vector’s population dynamic and could be used to develop novel control strategies.In the present study, an intensive search for the breeding sites of Lutzomyia longipalpis was conducted in urban and peri-urban areas of two municipalities, Promissão and Dracena, which are endemic for visceral leishmaniasis in São Paulo State, Brazil. During an exploratory period, a total of 962 soil emergence traps were used to investigate possible peridomiciliary breeding site habitats such as: leaf litter under tree, chicken sheds, other animal sheds and uncovered debris. A total of 160 sand flies were collected and 148 (92.5%) were L. longipalpis. In Promissão the proportion of chicken sheds positive was significantly higher than in leaf litter under trees. Chicken shed habitats presented the highest density of L. longipalpis in both municipalities: 17.29 and 5.71 individuals per square meter sampled in Promissão and Dracena respectively. A contagious spatial distribution pattern of L. longipalpis was identified in the emergence traps located in the chicken sheds.The results indicate that chicken sheds are the preferential breeding site for L. longipalpis in the present study areas. Thus, control measures targeting the immature stages in

chicken sheds could have a great effect on reducing the number of adult flies and consequently the transmission rate of Leishmania (Leishmania) infantum chagasi.

- EPIDEMIOLOGY [562] P 749 - A WHOLE BLOOD IFN- Y / IL-10 RELEASE ASSAY AS A MARKER OF THE CELLULAR IMMUNE RESPONSE TO LEISHMANIA DONOVANI INFECTION IN HEALTHY INDIVIDUALS SINGH, O.P.1; SINGH, A.K.1; SINGH, N.1; GIDWANI, K.1; CHOURASIA, A.1; HASKER, E.2; SINGH, R.P.1; MCLEAN, D.3; JONES, S.L.3; BOELAERT, M.2; SUNDAR, S.1 1.DEPARTMENT OF MEDICINE, INSTITUTE OF MEDICAL SCIENCES, BANARAS HINDU UNIVERSITY, VARANASI, INDIA; 2.DEPARTMENT OF PUBLIC HEALTH, INSTITUTE OF TROPICAL MEDICINE, ANTWERP, BELGIUM; 3.CELLESTIS LIMITED, CHADSTONE, VICTORIA, AUSTRALIA.

Keyword:ifn-

/ il-10 release assay ; sla, dat, elisa

Abstract: Background: In an area endemic for visceral leishmaniasis (VL), asymptomatically infected persons play crucial role in transmission of disease. We have recently developed a whole blood IFN- / IL10 release assay as a marker of cellular immunity to detect infected and non- infected persons with high accuracy. In the present study, we evaluated this test amongst healthy individuals living in the kala-azar endemic zone in Bihar, India for identification of individuals exposed to leishmania infection but without disease.

Methods: We enrolled 13,163 persons from eleven highly endemic villages to identify the incident infected healthy persons as measured by seroconversion with DAT and rK39 ELISA at base line survey and at 12 months interval. Whole blood assay was performed longitudinally for two years amongst seropositive and its matched seronegative controls populations using Soluble Leishmania Antigen (SLA). IFN- and IL-10 were determined in antigen stimulated whole blood culture supernatant by conventional ELISA. Subjects were followed up on monthly basis to monitor the progression into disease.

Results: Of 13,163 persons only 309 subjects (3.6%) had converted to seropositive on either of the DAT or rK39 ELISA in one year interval. The percentage of persons with an IFN- -positive response (> 0.78 IU/mL) was equal in both seropositive (19%) and its control groups (16.3%).

Of those IFN- -positive persons 63.8% and 60.7% person remain IFN- positive over 1 year in seropositive and control groups respectively. The new incidence of IFN- positivity in healthy persons was 5.31%. Only one subject developed active disease that had high IFN- and IL-10 levels at base line.

Conclusion: These findings confirm that SLA based IFN- / IL-10 release assay is a promising tool for identification of clinically exposed, immune individuals.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [564] P 750 - SUCEPTIBILITY OF PHLEBOTOMUS ORIENTALIS AND P.BERGEROTI TO INSECTICIDES FROM ETHIOPIA MANAGIDO, M.B.; TSEGAYE, I.; GEBRE-MICHAEL, T. AKLILU LEMMA INSTITUTE OF PATHOBIOLOGY, ADDIS ABABA UNIVERSITY, ADDIS ABABA, ETHIOPIA.

Keyword:p.orientalis,p.bergeroti; insecticides; vector control

Abstract: Phlebotomus orientalis is considered as the vector of Leishmania donovani in Ethiopia. P. bergeroti is a suspected vector of L. major elsewhere in Africa and experimental studies have also proven this possibility. It is also suspected to transmit sandfly fever in Ethiopia. In the past, no effort has been made to control sandfly vectors by the application of insecticide based tools in the country. However, exposure to insecticides used for malaria vector control such as indoor residual spraying of insecticides and long lasting insecticidal nets as well as to agricultural pesticides might be inevitable and induce resistance in sandflies. In order to elucidate the status of insecticide susceptibility of the two species, WHO susceptibility tests were conducted on populations of P.orientalis from three localities, Melka-Worer in the Awash Valley in the east, Libo-Kemkem in the north and Metema in northwest. P.bergeroti was tested from Melka-Worer only. The insecticides included one organochlorine (DDT), one orgnophosphate (malathion) and four pyrethroids (permethrin, lambdacyhalothrin, deltamethrin and alphacypermethrin) and tests were conducted on either F1 or wild caught females. The KDT50, KDT95 and mortality was determined. The results showed that the KDT50 and KDT95 of the pyrethroids to P.orientalis from Melka-Worer and Libo-Kemkem remained under 13 and 23 minutes except lambdacyhalothrin in the latter locality in which 18 and 27 minutes were recorded. The Metema population appeared to be less affected as the values of KDT50 ranged from 21.5-28.9 and KDT95 from 31.7-45.7 minutes. On the other hand, the KDT50 of DDT in all places was from 20 to 27 minutes. In terms of mortality, P. orientalis was greatly susceptible to all insecticides (mortality 98-100%) except for malathion from Melka-Worer in which percentage mortality was 93.8. Similarly, P.bergeroti was found to be significantly susceptible to all insecticides as mortality was 100%. However, the KDT50 and KDT95 values of the pyrethroids were greater than those of P.orientalis. In conclusion, both species are susceptible to the tested insecticides, and therefore, vector control in peridomestic habitats using one of these insecticides is warranted in Ethiopia.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [566] P 751 - VECTORS OF THE LEISHMANIASES IN ETHIOPIA GEBRE-MICHAEL, T.1; MANAGIDO, M.B.1; MEKURIA, A.H.2; WARBURG, A.3

1.AKLILU LEMMA INSTITUTE OF PATHOBIOLOGY , ADDIS ABABA UNIVERSITY, ADDIS ABABA, ETHIOPIA; 2.. COLLEGE OF HEALTH SCIENCES, ADDIS ABABA UNIVERSITY, ADDIS ABABA, ETHIOPIA; 3.THE KUVIN CENTER FOR THE STUDY OF TROPICAL AND INFECTIOUS DISEASES,THE HEBREW UNIVERSITY-HADASSAH M, JERUSALEM, ISRAEL.

Keyword:phlebotomus ; visceral leishmaniasis, cutaneous leishmaniasis, ; ethiopia

Abstract: Teshome Gebre-Michael1, Meshesha Balkew1, Asrat Hailu2 and Alon Warburg3 1. Aklilu Lemma Institute of Pathobiology , Addis Ababa University, P. O. Box 1176, Addis Ababa, Ethiopia, 2. College of Health Sciences, Addis Ababa University, P. O. Box 1176, Addis Ababa, Ethiopia, 3.The Kuvin Center for the Study of Tropical and infectious Diseases,The Hebrew University-Hadassah Medical School Jerusalem 91120, ISRAEL Abstract Information on the vectors of the leishmaniases in Ethiopia is reviewed. To date, there are least 19 species of Phlebotomus belonging to six subgenera, of which at least eight species are proven vectors of various species Leishmania. Two species, P. (Larroussius) orientalis and P. (Synphlebotomus) martini are the principal vectors of visceral leishmaniasis (VL, L. donovani)

in southern endemic areas of Ethiopia, from which the parasite has been isolated and typed. The former species is also the suspected vector in the much larger and most important endemic areas of northern and northwest of Ethiopia, but the parasite has yet to be isolated/detected. P. (S.) celiae and P. (Anaphlebotomus) rodhaini, sympatric with P. martini in the south while, the latter, with P. orientalis in the north are considered secondary vectors since L. donovani has been detected from few specimens of both species in the south, requiring further investigation.

The vectors of cutaneous leishmanisis (CL) caused by L. aethiopica and L. major and L. tropica are various. L. aethiopica which is the most important and widespread cause of CL in the Ethiopian highlands (1700-2700masl) is vectored by three species belonging to two subgenera Larroussius (P. longipes and P. pedifer) and Paraphlebotomus (P. sergenti). However, the parasite was isolated and typed from the latter species in a low altitude Rift Valley area (1000 masl) of eastern Ethiopia where human or animal infections are apparently absent. In addition, P. (Pa.) sergenti and P. (Pa.) saevus) have been found vectors of L. tropica, in the eastern Ethiopia (Rift Valley Region), though CL due to L. tropica is so far rare in Ethiopia. P. (Phelobotomus) duboscqi is a vector of L. major (CL) in southern Ethiopia, where the parasite is similarly so far rare in Ethiopia. Parasites have also been isolated/detected from P. (La.) ashfordi and P. (Adlerius) arabicus, but remain unidentified.

- EPIDEMIOLOGY [581] P 752 - LEISHMANIA PARASITE LOAD STUDY BY QPCR: THRESHOLD VALUE FOR ASYMPTOMATIC INFECTION DETECTION IN KALA AZAR ENDEMIC REGION MUZAFFARPUR, IN INDIA. SUDARSHAN, M.; SUNDAR, S. BANARAS HINDU UNIVERSITY, VARANASI, INDIA.

Keyword:leishmania; qpcr; muzaffarpur,india

Abstract: Background- Case under estimation or misdiagnosis can lead to lethal condition in Kala azar.

Infection detecting tool should make the distinction between acute disease,cure status and asymptomatic infection. This work represent qPCR as a tool for detection as well as quantification of Leishmania donovani in individuals from endemic region of Visceral leishmaniasis (VL), i.e., Muzaffarpur, Bihar.

Material and Method- Peripheral blood collected in citrate tubes from different subjects are patients(n=40), past case history (cure)(n=94) , endemic healthy (EHC)(n=130) as well as nonendemic healthy(40). Leishmania specific kDNA primer is used for parasite quantification.

Result- 36.9% positivity is shown by SYBR green based qPCR in endemic healthy. If detected, qPCR show Parasitemia level in endemic healthy as well as past case history of median value of less than 1parasite genome/ml of blood, having range is 0.008-5.49 while in Patients it is 554,610 parasite genome/ml of blood.

Conclusion- Parasite load in different samples determine threshold level as near to 5 parasite genome/ml of blood are on the risk of clinical outcome of infection,i.e, symptomatic. This study will be helpful for the assesment of the endemic persons for detection of parasite load and to categorise them for threshold reference values in cryptic infections as asymptomatic to decide their contribution in disease transmission and their further follow up as progressor or non progressor.

- CONTROL PROGRAMS [588] P 753 - EVIDENCE ON INSECTICIDE TREATED BED NETS TO CONTROL VISCERAL LEISHMANIASIS IN THE INDIAN SUB-CONTINENT CHOWDHURY, R.1; BHANDERI, P.K.2; CHOWDHURY, V.3; FARIA, S.1; BOELAERT, M.4; DASH, A.P.5

1.NATIONAL INSTITUTE OF PREVENTIVE AND SOCIAL MEDICINE (NIPSOM), DHAKA, BANGLADESH; 2.ACI FORMULATION LIMITED, DHAKA, BANGLADESH; 3.DHAKA COLLEGE, DHAKA, BANGLADESH; 4.INSTITUTE OF TROPICAL MEDICINE, ANTWERP, BELGIUM; 5.WHO REGIONAL OFFICE FOR SOUTH-EAST ASIA, NEW DELHI, INDIA.

Keyword:visceral leishmaniasis, vector control; insecticide treated nets, indian subcontinent; elimination, phlebotomus argentipes

Abstract: In the Indian sub-continent (ISC), Visceral Leishmaniasis (VL) is caused by Leishmania donovani, and transmitted by Phlebotomus argentipes. Bangladesh, India and Nepal are working for elimination of VL by 2015 and integrated vector management (IVM) is one of the important elements for that. Indoor residual spraying (IRS) is very effective to reduce peridomestic vector density but it requires strict monitoring and supervision which is not easy to ensure by the national programmes. The use of insecticide treated nets (ITNs) is a potential alternative for sand fly control as bed net use is a common practice in the endemic regions. ITNs combine the individual protection of a bed net with the effect of insecticide. We systematically reviewed the evidence available about the effect of bed nets and ITNs on VL in the ISC. A study in Nepal showed an association between regular use of bed net and protection from VL (OR 0.23, P < 0.001) but another study could not find protective effect. A study in Bangladesh showed consistent use of bed nets especially during summer was protective (OR 0.69, P = 0.01).

The efficacy of ITNs tested in a study to prevent L. donovani infection and VL in Nepal and India where ITNs significantly reduced P. argentipes density per house by 24.9%. However, there was no significant difference found in the risk of seroconversion over 24 months. A study in ISC, showed in pooled analysis a 43.7% reduction of sand fly density by ITNs and only in Bangladesh IRS and ITNs were associated with a 70–80% decrease of P. argentipes density up to 5 months post intervention. Vector density rebounded by 11 months post-IRS, whereas ITNtreated households (HH) continued to show significantly lower density compared with HHs without intervention. Another study in Bangladesh has evaluated the effect of ITNs and showed reduced sand fly density by 60% at 18 months post-intervention.

In summary, the evidence so far points to a marked to moderate effect of ITNs on sandfly density in the ISC, with possibly a more pronounced effect in Bangladesh than in India and Nepal. There is no evidence so far, ITN would have an additional impact on L. donovani transmission in the current context of VL endemic communities in India and Nepal, with high rates of commercial bed net use and IRS of irregular quality. ITNs should be considered as an additional tool in an IVN strategy without neglecting IRS. More research on alternative and effective sand fly control tools is needed.

- EPIDEMIOLOGY [597] P 754 - PHLEBOTOMINIC FAUNA AND NATUAL INFECTION BY LEISHMANIA SPP. IN THE MUNICIPALITY OF NOVA MUTUM, MATO GROSSO, BRAZIL THIES, S.F.1; MARIA, A.L.2; MICHALSKY, É.M.3; MIYAZAKI, R.D.4; FERNANDES, C.J.5; DIAS, E.S.3 1.MEDICAL ENTOMOLOGY LABORATORY, SCHOOL OF MEDICAL SCIENCESS, UFMT/MATO GROSSO STATE SECRETARY/SES., CUIABÁ, MT, BRAZIL; 2.MEDICAL ENTOMOLOGY LABORATORY, SCHOOL OF MEDICAL SCIENCESS, UFMT, CUIABÁ, MT, BRAZIL; 3.LEISHMANIASIS LABORATORY, RENÉ RACHOU RESEARCH CENTRE/FIOCRUZ, BELO HORIZONTE, MG, BRAZIL; 4.BIOLOGY AND ZOOLOGY, INSTITUTE OF BIOSCIENCES, UFMT, CUIABÁ, MT, BRAZIL; 5.DEPARTAMENT OF MEDICAL PRACTICE, SCHOOL OF MEDICINE, JÚLIO MÜLLER UNIVERSITY HOSPITAL, CUIABÁ, MT, BRAZIL.

Keyword:leishmania; nova mutum; l. antunesi

Abstract: The municipality of Nova Mutum is situated in the mid-north region of Mato Grosso state, which in recent years has reported cases of American Tegumentary Leishmaniasis (ATL) in humans. In the rural area one specific place (woodland), has presented human cases with common aspects, such as short incubation period and multiple lesions. The objective of this work was to identify phlebotominic fauna and verify natural infection by Leishmania spp. in a rural area of the municipality of Nova Mutum, Mato Grosso. Phlebotamine sand flies were captured every other month from June 2011 to April 2012 using luminous CDC traps between 17:00 and 07:00 the next day for three consecutive nights, arranged at 10 equidistant points, roughly every 100 metres from the edge into the heart of the forest. The captured specimens were identified according to the Young and Duncan’s proposed classification (1994). Some pools containing between 1 and 10 females were formed, separated by species, the trap number and capture date. The Polymerase Chain Reaction (PCR) method was used to extract and detect Leishmania DNA. The results showed that an extremely diverse fauna, with 31 different species captured, one of the Brumptomyia genus and 30 of the Lutzomyia genus. The most abundant species was L. antunesi (45,44%), followed by L. saulensis (20,57%) which were found every month of collection and at every capture point. 73,15% of the specimens were female. The most prolific month for collection was October 2011. Others species of importance in ATL epidemiology were collected, such as L. flaviscutellata, L. whitmani and L. umbratilis. The highest number of captured phlebotomine sand flies occurred in months of the rainy season (October and December), when the relative humidity and temperature were higher. Thirteen pool naturally infected by Leishmania spp. were found, corresponding to a general infection rate of 4,44% (13/293). The L. antunesi and L. ubiquitalis were found to be naturally infected by Leishmania spp. with rates varying from 5,61% (11/196) to 12,5% (2/16), respectively. The natural infection allied to the abundance in the area investigated suggest the participation of L. antunesi and L. ubiquitalis, in the ATL zoonotic cycle in Nova Mutum, Mato Grosso.

- EPIDEMIOLOGY [602] P 755 - PHLEBOTOMINIC FAUNA IN THE FEDERAL UNIVERSITY OF MATO GROSSO (UFMT) CAMPUS AND SURROUNDING NEIGHBOURHOODS IN CUIABÁ, MATO GROSSO THIES, S.F.1; MARIA, A.L.2; RODRIGUES, J.S.2; LEITE, H.S.3; RAMOS, S.R.3; GONÇALVES, E.L.4; MIYAZAKI, R.D.5 1.MEDICAL ENTOMOLOGY LABORATORY, SCHOOL OF MEDICAL SCIENCESS, UFMT/MATO GROSSO STATE SECRETARY/SES., CUIABÁ, MT, BRAZIL; 2.MEDICAL ENTOMOLOGY LABORATORY, SCHOOL OF MEDICAL SCIENCESS, UFMT, CUIABÁ, MT, BRAZIL; 3.REGIONAL HEALTH OFFICE OF BAIXADA CUIABANA, SES, CUIABÁ, MT, BRAZIL; 4.ZOONOSES CONTROL CENTRE, CUIABÁ MUNICIPAL HEALTH SECRETARY, CUIABÁ, MT, BRAZIL; 5.DEPARTAMENT OF BIOLOGY AND ZOOLOGY, INSTITUTE OF BIOSCIENCES, CUIABÁ, MT, BRAZIL.

Keyword:leishmaniases; phlebotomine; ufmt

Abstract: Leishmaniases are zoonotic diseases that can afflict man, transmitted through insects of the Lutzomyia genus, denominated phlebotominae, commonly known as sand flies. The disease is manifested in two forms: American tegumentary leishmaniasis, which afflicts the skin and mucous membranes, and visceral leishmaniasis, which afflicts the spleen, liver and other internal organs. The aim of this study was to monitor the presence of phlebotomine sand flies of veterinary medical relevance on the campus of the Federal University of Mato Grosso and to detect their presence in six local neighbourhoods around the university campus. Monitoring was conducted monthly, from December 2008 to December 2010, with the installation of CDC light

traps at six points around the campus for three consecutive nights from 6:00 p.m. until 6:00 a.m. the following day. The surrounding neighbourhoods monitored were Santa Cruz, Morada dos Nobres, Boa Esperança, Jardim das Américas, Jardim Itália and Renascer, employing the same collection technique, from August to October 2010. Following the collections, the insects were prepared and identified in the Entomology Laboratory of UFMT and the State Secretary of Health, according to guidelines set by Young and Duncan (1994). Thirteen species belonging to the Lutzomyia genus and specimens of Brumptomyia were captured on the UFMT campus, totalling 436 insects, of which 83.03% were found to be of veterinary medical relevance, broken down as follows: L. cruzi (267), L. whitmani (87), L. longipalpis (7) and 1 of the longipalpis complex, 66,57% were female and 33,43% male. In the surrounding neighbourhoods, seven species were found belonging to the Lutzomyia and specimens of Brumptomyia, totaling 42 specimens, of which 15 (35,71%) were of veterinary medical relevance; the presence of L. whitmani was detected in the neighbourhoods of Santa Cruz (1), Boa Esperança (7), Morada dos Nobres (1), Jardim das Américas (1) and insects of the longipalpis complex in Boa Esperança (1), Morada dos Nobres (2) and Santa Cruz (1), of which 42,86% were female and 57,14% male.

Considering the abundance and diversity of the findings, an active entomological, epidemiological and environmental surveillance program is recommended to ensure early detection of environmental changes that could alter the population density and diversity of the sand flies and any emergence of new cases of leishmaniasis, thus enabling the adoption of preventive and curative measures, when required.

- EPIDEMIOLOGY [608] P 756 - ANALYSIS OF THE EPIDEMIOLOGICAL SITUATION OF VISCERAL LEISHMANIASIS IN THE MUNICIPALITY OF ARAGUAÍNA (TO): PERIOD 2005-2009 TOLEDO, C.R.S.; SOBRAL, A.A.; SABROZA, P.C.; CHAVES, S.A.M. ENSP/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:visceral leishmaniasis; dissemination pattern; araguaína, epidemiology

Abstract: The objective of this study was to describe the epidemiological situation of human visceral leishmaniasis (VL) in the urban area of the municipalIty of Araguiana (TO) during the period 2005-2009 in order to understand the pattern of cases dissemination in relation with the social organization from the time-space distribution of the disease. The analysis of the human VL spatial and temporal distribution was correlated with the socio demographic situation and the situation of urban infrastructure, in order to characterize and identify the areas of major vulnerability for the occurrence of the disease in the municipality. For the epidemiological characterization of human cases, we used variables included in the Information System for Notifiable Diseases (SINAN) data bank, exclusively relative to habitants of the municipality of Araguiana: sex, age, co-infection HIV/AIDS and outcome. These variables were assessed by using proportions and specific rates. The confirmed autochtonous cases of human VL were georefrenced based on the address found in SINAN data bank. The study used data from sample surveys conducted canines by the Municipal Health Araguaína to estimate the positive canine in the city between 2006 and 2008. During the study period, a sharp increase in the number of confirmed human VL cases (341.6%) was observed in the municipality with a pattern of diffuse peri-urban dissemination occurring, predominantly, in the urban periphery and, secondarily, in areas central/urbanized. We found a predominance of cases in the age class 0-4 years (49.4%), in agreement with the epidemiological pattern observed in the North Eastern region of Brazil.

The main symptoms observed were: fever, asthenia and weight loss. From the survey data canines sampling we observed little change in the positivity of the canine VL in the years 2006 (33.51%), 2007 (37.88%) and 2008 (37.12%) even with the gradual increase in the number of samples collected, respectively, 1549, 4728 and 6223. The analyses reinforced the hypothesis that the occurrence of VL is closely related to the situation of social vulnerability.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [612] P 757 - SEASONAL DYNAMICS, RATES OF INFECTION, AND HOST-FEEDING PREFERENCES OF PHLEBOTOMUS PERNICIOSUS IN A NEW FOCUS OF LEISHMANIASIS IN MADRID, SPAIN. JIMÉNEZ, M.1; GONZÁLEZ, E.1; HERNÁNDEZ, S.1; IRISO, A.2; FÚSTER, F.2; MOLINA, R.1

1.UNIDAD DE ENTOMOLOGÍA MÉDICA, SERVICIO DE PARASITOLOGÍA, CENTRO NACIONAL DE MICROBIOLOGÍA, ISCIII, MAJADAHONDA, MADRID, SPAIN; 2.SUBDIRECCIÓN GENERAL DE SANIDAD AMBIENTAL, CONSEJERÍA DE SANIDAD, COMUNIDAD DE MADRID, MADRID, SPAIN.

Keyword:leishmaniasis; sand flies; madrid

Abstract: Since 2010 it has increased the number of cases of both human visceral leishmaniasis and cutaneous leishmaniasis in southwestern Madrid region (Spain). Direct xenodiagnosis of leishmaniasis in hares (Lepus granatensis) from the focus of the disease proved that they are infective to colonized Phlebotomus perniciosus. Further characterization of promastigotes isolated from these sand flies fed on hares demonstrates that these lagomorphs were infected by Leishmania infantum. Surprisingly the prevalence of canine leishmaniasis in the area of the focus is even lower than that detected in the neighboring areas. In a previous entomological survey carried out in the same area in 2011 P. perniciosus was the only one potential vector identified. In order to a better understanding of the new active focus an entomological survey was carried out from May to October 2012 in 4 stations placed neighbouring the urban area and selected according the high number of sand flies collected in 2011 in the same area. Each month 20 sticky traps (20x20 cm) and 2 CDC traps were used during two consecutive nights in every season. CDC traps were replaced every day. The main objective was to establish the phenology of P. perniciosus in the focus and its relationship to the rates of infection of this species.

Additionally we attempted to identify the source of the blood ingested by females. A total of 16181 specimens were collected, 7709 sand flies by CDC light traps and 8472 in sticky traps.

Taxonomical identification is in progress. For now three sand fly species have been identified: P. perniciosus (85.3%), P. sergenti (0.03%), and S. minuta (14.6%). 735 P. perniciosus female collected with CDC traps in the 4 stations of seasonal study were dissected and 18 of them were found infected with L. infantum (2.45%). Infected females were found in the four stations during June, July, and August. A total of 14 isolates were obtained and characterized as L. infantum.

Host-feeding preferences of sand flies in wild habitats is being done by PCR of cytochrome b, sequencing, and subsequent comparison with sequences deposited in the GenBank database using standard nucleotide BLAST searches. The following hosts have been identified for now: hare, rabbit, dog, and human. The data presented here provide significant epidemiological information relative to the spread of human leishmaniasis in the focus. This study was funded by EU grant GOCE-2003-010284 EDENext and Comunidad Autónoma de Madrid, Spain.

- EPIDEMIOLOGY [615] P 758 - PHLEBOTOMINE SAND FLY (DIPTERA: PSYCHODIDAE) FAUNA IN THE MUNICIPALITY OF JABOTICATUBAS, MINAS GERAIS STATE, BRAZIL – PRELIMINARY RESULTS LANA, R.S.1; MONTEIRO, É.M.1; SILVA, J.C.F.2; SILVA, F.O.L.1; COSTA, A.S.N.3; DIAS, E.S.1 1.CENTRO DE PESQUISAS RENÉ RACHOU/FUNDAÇÃO OSWALDO CRUZ, BELO HORIZONTE, MG, BRAZIL; 2.INSTITUTO DE CIÊNCIAS BIOLÓGICAS, UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 3.SECRETARIA DE SAÚDE, JABOTICATUBAS, MG, BRAZIL.

Keyword:fauna; lutzomyia; jaboticatubas

Abstract: Jaboticatubas is a Brazilian municipality located in the Serra do Espinhaço at 100 km of the capital of the state of Minas Gerais. It is an ecotourism destination since it holds 80% of the total area of the National Park of Serra do Cipó, an ecological sanctuary with rich and diversified fauna and flora. So far, the phlebotomine sand fly fauna there is unknown. Due to recent notifications of human cases of tegumentary and visceral leishmaniases in the municipality we started an entomological survey aiming to describe the local phlebotomine sand fly fauna and to determine their seasonal fluctuation, the natural rate of Leishmania infection, besides identifying the circulating Leishmania in the sand flies infected, if any. Field captures started on May 2012 and are planned to last on April of 2013. HP light traps are mounted for three consecutive nights per month in eight districts of the urban area and two districts in the countryside of Jaboticatubas. In the first six months of our study, a total of 463 sand flies were captured. Seven distinct species were identified under the following percentages: L. cortelezzi (4.3%), L. intermedia (20.9%), L. lenti (2.2%), Lutzomyia longipalpis (12.7%), L. migonei (0.6%), L. pessoai (5.8%) and L. whitmani (52.9%). The high abundance of phlebotomine sand flies species that are proven vectors of leishmaniases accounted for 87.1% of the captured specimens.

Work is in progress to achieve the objectives proposed.

Financial support – FAPEMIG, CNPq and FIOCRUZ

- RESERVOIRS [618] P 759 - STUDIES ON THE CANINE VISCERAL LEISHMANIASIS IN BELO HORIZONTE, AN ENDEMIC CAPITAL IN THE STATE OF MINAS GERAIS, BRAZIL. MICHALSKY, E.M.1; LARA-SILVA, F.O.1; SILVA, M.A.1; COSTA, A.J.1; FRANÇA-SILVA, J.C.2; GOMES, L.1; LIMA, A.C.1; FIUZA, V.O.3; PAGLIONI, D.N.3; ASSIS, A.P.3; BUSSOLOTTI, A.S.3; FORTES-DIAS, C.L.4; DIAS, E.S.1 1.CPQRR, BELO HORIZONTE, MG, BRAZIL; 2.UFMG, BELO HORIZONTE, MG, BRAZIL; 3.SECRETARIA DE SAUDE, BELO HORIZONTE, MG, BRAZIL; 4.FUNED, BELO HORIZONTE, MG, BRAZIL.

Keyword:canine visceral leishmaniasis; belo horizonte; prevalence rate

Abstract: The spreading of the visceral leishmaniasis (VL) in Belo Horizonte, the capital of Minas Gerais state, illustrates well the urbanization of the disease. Since 2001, increasing numbers of human cases of VL have been reported in the city, with 277 human cases between 2009 and 2010. The first human cases were preceded by canine cases and the overall prevalence rate of Leishmania infection in dogs was around 7%, in the same period. In the present work we aim to determine the current status of canine VL in two districts of Belo Horizonte, named Miramar and Salgado Filho. Canine surveys were performed from 2010 to 2012 and Leishmania infection was assayed by enzyme-linked immunosorbent (ELISA) and immunofluorescence (IFA) assays. A total of 4,328 dogs were examined with an average of 721 animals per year and district. The prevalence rates of Leishmania infection per year were determined as 8.1% (2010), 3.7% (2011), 2.3% (2012) for the Miramar district and 4.8% (2010), 2.2% (2011), 4.2% for the Salgado Filho district. After necropsy, tissue fragments from the spleen, bone marrow, lymph nodes and skin from 51 seropositive dogs were tested for the presence of Leishmania parasites by nested PCR (LnPCR) targeting the SSUrRNA gene. The overall percentages of positive samples were as follows: 100.0% for the spleen, 80.4% for the bone marrow, 56.9% for the lymph nodes and 68.6% for the skin, respectively. Bone marrow aspirates were seeded in NNN/LIT culture medium and thirty-eight samples (74.5%) showed Leishmania growth. After subculturing, the promastigotes biomass will be submitted to nested PCR (LnPCR). Gel-extracted amplicons from

both bone marrow subcultures and tissue samples will be sequenced aiming to identify the circulating Leishmania species in Belo Horizonte.

Financial support: FAPEMIG, FIOCRUZ, Secretaria Municipal de Saúde de Belo Horizonte, CNPq.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [619] P 760 - PHLEBOTOMINE SAND FLIES (DIPTERA, PSYCHODIDAE) COLLECTED IN THE SURUWAHÁ INDIAN RESERVE, AMAZONAS STATE, BRAZIL ASSIS, M.D.G.G.; BARROS, J.A.C.; SHIMABUKURO, P.H.F. FIOCRUZ, BELO HORIZONTE, MG, BRAZIL.

Keyword:amazon; indigenous people; phlebotominae

Abstract: The Suruwahá are a small, isolated indigenous population of approximately 140 individuals living in the southern Amazon. The Suruwahá live on agriculture and fishing, while hunting is the main and most prestigious activity. Since 2010 there has been an increase in the number of American cutaneous leishmaniasis (ACL) cases recorded among the Suruwahá, which seems to be related to the distribution of flashlights to the community by the Fundação Nacional do Índio (FUNAI). The use of flashlights has caused a shift in the hunting activity of the Suruwahá from strictly diurnal to nocturnal, thus increasing their exposure to the sand fly vectors of Leishmania which bloodfeed at dusk/night. Among the Suruwahá, leishmaniasis is called “idu”, which is also the name of a foul smelling forest mushroom, that resembles the shape of the ACL lesion, and is believed to harbor a spirit that throws arrows that cause the disease. The objective of this work is to study the sand fly fauna collected in the Suruwahá Indian Reserve, Tapauá municipality, Amazonas state, Brazil. The sand flies were collected during a training course on leishmaniasis at the FUNAI Suruwahá base in February 2012. Two CDC light traps were placed at hunting tracks and operated for 3 consecutive nights. We collected 374 sand flies (247 males and 127 females). The most common species was Trichophoromyia ubiquitalis (82%), while the remaining specimens were identified as Th. octavioi, Th. melloi and Th. brachipyga. The sand fly species Th. ubiquitalis has a wide geographical distribution in the Amazon and is an anthropophilic species that has been incriminated in the transmission of Leishmania (Viannia) lainsoni in Pará state. Further studies are necessary to assess the diversity of sand flies in this area in order to understand the transmission dynamics of Leishmania among the Suruwahá.

Financial support: CNPq grant no. 474506/2012-6.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [620] P 761 - FREQUENCY OF SANDFLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN ADRIANOPLE, A TRANSMISSION AREA OF AMERICAN CUTANEOUS LEISHMANIASIS IN THE STATE OF PARANA, BRAZIL. GONÇALVES, A.L.1; CASTRO, E.A.1; SILVA, C.P.C.1; MELO, A.L.A.1; DE SOUZA, N.A.2; THOMAZ-SOCCOL, V.1 1.UNIVERSIDADE FEDERAL DO PARANÁ, CURITIBA, PR, BRAZIL; 2.FUNDAÇÃO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:american cutaneous leishmaniasis; phlebotominae sand flies; paraná

Abstract: American cutaneous leishmaniasis (ACL) represents a serious public health problem, with repercussions on the economy of endemic countries. Unregulated construction and development by human populations increases the risk of peridomestic transmission and characterizes a new transmission profile. The diversity of sandfly vectors and reservoirs of dermotropic Leishmania species is key improving current understanding of the dynamics of disease transmission. To determine the likely time that the population is more susceptible to being bitten and to acquiring ACL, the hourly rate of sandflies (Psychodidae: Phlebotominae) was investigated in October 2012 in the city of Adrianople, an area endemic for Leishmania (Viannia) braziliensis in the State of Paraná, Brazil. Shannon traps were installed in peridomiciles for one night, between 6 pm and 6 am. In this preliminary assessment, 878 insects were collected. The times of greatest frequency of collection of sandflies were between 3 and 4 am (122 insects collected), midnight and 1 am and 5 and 6 am (120), while fewer insects were collected between 6 and 7 pm (04), 7 and 8pm (28) and between 2 and 3 am (24). The most abundant species was Lutzomyia intermedia s.l., 851 examples collected, followed by Lutzomyia fischeri (15) and Lutzomyia migonei (12). Lutzomyia intermedia s.l. was more abundant during the second half of the night, 547 examples collected. This species predominated between midnight and 1 am and between 3 and 4 am. All the examples of Lu. fischeri were collected between 11 pm and 6 am, while Lu. migonei was also collected more frequently during the second half of the night. The abundance of specimens of Lu. intermedia confirms its dominance in the region and its possible role as a Leishmania vector. Lutzomyia fisheri and Lu. migonei should be considered possible vectors, because they have been found parasitized in other endemic areas in Brazil; however, due to their low density, the vector role is secondary. Studies developed in Adrianople strongly suggest the need to adopt policies that involve health education actions, associated with the notion of environmental management and basic concepts concerning the disease as part of a successful integrated program of entomological surveillance and ACL control.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [623] P 762 - SAND FLIES COLLECTED IN THE CAITITU INDIAN RESERVE (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE), AMAZONAS STATE, BRAZIL SILVA, T.R.R.1; ASSIS, M.D.G.G.2; FREIRE, M.P.1; GONTIJO, C.M.F.2; SHIMABUKURO, P.H.F.2 1.PPG-SSEA, MANAUS, AM, BRAZIL; 2.FIOCRUZ, BELO HORIZONTE, MG, BRAZIL.

Keyword:amazon; leishmaniasis; vectors

Abstract: Sand flies collected in the Caititu Indian Reserve (Diptera: Psychodidae: Phlebotominae), Amazonas state, Brazil Túllio Romão Ribeiro da Silva1, Mauro Diego Gobira Guimarães de Assis 2, Maíra Posteraro Freire1, Célia Maria Ferreira Gontijo3, Paloma Helena Fernandes Shimabukuro2 1Programa de Pós-Graduação em Saúde, Sociedade e Endemias na Amazônia, Universidade Federal do Amazonas/Instituto Leônidas e Maria Deane/Universidade Federal do Pará; 2Centro de Referência Nacional e Internacional para Flebotomíneos, Centro de Pesquisas René Rachou, FIOCRUZ/MG; 3Laboratório de Leishmanioses, Centro de Pesquisas René Rachou, FIOCRUZ/MG

American cutaneous leishmaniasis (ACL) in Lábrea municipality is mainly associated with activities such as extraction of Brazil nuts, rubber, copaiba oil, among other forest products, and affects both indigenous and non-indigenous populations. Data on ACL in indigenous populations in this area are scarce, and there is even less information on the etiological agents, the non-human animal reservoir hosts maintaining zoonotic/sylvatic transmission, and the sand fly vectors involved. The objective of this work was to study the fauna of sand flies collected in the Caititu Indian Reserve, Lábrea municipality, Amazonas state, Brazil. The insects were collected during seven nights in February 2012, along the Transamazônica road (BR-230). Sand flies were collected, using 10 CDC light traps placed at the Brazil nut collection sites (“piques de castanha”) used by the indigenous people, during the period of nut collection. The insects were identified to species level using morphology, and DNA extraction from pooled females followed by ITS1 PCR were used to detect Leishmania DNA in these samples. A total of 960 specimens distributed in 11 genera and 25 species were collected. The most abundant species were Psychodopygus davisi (21% of collected sand flies), Nyssomyia antunesi (12%) and Trichophoromyia ubiquitalis (8%). All three species have been previously implicated in the transmission of Leishmania parasites: Le. braziliensis and Le. naiffi, Le. lindenbergi and Le. lainsoni, respectively. A total of 524 females distributed in 78 pools were used for the molecular detection of Leishmania, of which 11 (14%) pools were positive. Future DNA sequencing will provide information on the Leishmania species found in the studied sand flies. Our work confirms both the presence of Leishmania and sand fly vector species in the studied area. More studies are necessary to elucidate the role of each species in the transmission of ACL among the indigenous population of Lábrea municipality. Financial support: Projeto Saúde e Condições de Vida de Povos Indígenas na Amazônia, Programa de Apoio a Núcleos de Excelência – PRONEX/FAPEAM/CNPq, Edital 003/2009.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [625] P 763 - SALIVARY ANTIGENS FROM PHLEBOTOMUS ARGENTIPES: IDENTIFICATION AND ISOLATION MARTÍN-MARTÍN, I.; MOLINA, R.; JIMÉNEZ, M. UNIDAD DE ENTOMOLOGÍA MÉDICA, CENTRO NACIONAL DE MICROBIOLOGÍA, INSTITUTO DE SALUD CARLOS III, MADRID, SPAIN.

Keyword:phlebotomus argentipes; salivary proteins; sand fly

Abstract: In the Indian subcontinent visceral leishmaniasis is caused by the protozoa Leishmania donovani and is transmitted to humans by the bite of infected sand flies Phlebotomus argentipes in an anthroponotic cycle. Sand fly salivary proteins have been highlighted as vaccine candidates as sand fly saliva is involved in parasite establishment. Moreover, salivary proteins are gaining importance as markers of exposure. Therefore, studies on sand fly saliva would be useful to develop epidemiological tools that help to improve control strategies against leishmaniasis. We identified salivary antigens of P. argentipes by coupling two-dimensional electrophoresis and Western blot techniques. The apyrase SP03, the D7 related protein SP10, the antigen 5 SP05, the 33 kDa-like protein SP09, the PpSP15-like SP01, SP02 and SP07 and other proteins without assigned protein family (SP06, SP17, SP20 and SP56) were described as immunogenic proteins.

In a later step, a cDNA library constructed with 120 salivary glands from recently emerged P. argentipes was cloned into TriplEx2 vector and packaged into phage particles (Gigapack III Gold Packaging Extract, Agilent). The amplified library showed a titer of 1.72 x 1012 pfu/ml and a percentage of non recombinant clones of 5.96 as determined by plating the library by infecting log phase XL1-blue cells in the presence of IPTG and X-Gal. Several salivary antigens such as the apyrase SP03 (DQ136150), the yellow protein SP04 (DQ136151) and the 33 kDa-like

protein SP09 (DQ136155) among others were isolated from the cDNA library and successfully cloned into pCR®4-TOPO® vector (Invitrogen). To our knowledge, this is the first identification of antigens present in the saliva of P. argentipes. Moreover, a cDNA library of high titre and percentage of recombinant clones was successfully constructed and will be useful as a source of material for further studies. Several antigens were isolated from the cDNA library and will be further obtained as recombinant proteins to be tested as vaccines candidates or markers of exposure. This study was funded by the Spanish Ministry of Science & Innovation (Project AGL2008-01592) and EU grant GOCE-2003-010284 EDENext.

- RESERVOIRS [626] P 764 - SEROEPIDEMIOLOGY OF CANINE VISCERAL LEISHMANIASIS IN THE MUNICIPALITY OF JABOTICATUBAS, AN IMPORTANT ECOTOURISM DESTINATION IN THE STATE OF MINAS GERAIS, BRAZIL - PRELIMINARY RESULTS. LANA, R.S.1; SILVA, J.C.F.2; MONTEIRO, É.M.1; MARTINS, J.C.D.3; COSTA, A.S.N.3; MARCELINO, A.P.4; AVELAR, D.M.4; CARDOSO, F.A.4; COSTA, A.J.1; SILVA, K.M.S.1; DIAS, E.S.1

1.CENTRO DE PESQUISAS RENÉ RACHOU/FUNDAÇÃO OSWALDO CRUZ, BELO HORIZONTE, MG, BRAZIL; 2.INSTITUTO DE CIÊNCIAS BIOLÓGICAS, UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 3.SECRETARIA DE SAÚDE, JABOTICATUBAS, MG, BRAZIL; 4.FUNDAÇÃO EZEQUIEL DIAS, BELO HORIZONTE, MG, BRAZIL.

Keyword:canine; visceral; leishmaniasis

Abstract: Jaboticatubas is a municipality in the Brazilian state of Minas Gerais, with about 17.600 inhabitants. Jaboticatubas is an important ecotourism destination in the state since it holds 80% of the total area of the National Park of Serra do Cipó, an ecological sanctuary with rich and diversified fauna and flora. Taken into account recent notifications of human cases of visceral leishmaniases in the municipality and the epidemiological role played by dogs in the disease cycle, we focused the present study in canine VL. A census survey was performed with domiciled male and female dogs, from different ages and breeds, in five distinct districts of the municipality. Six hundred and sixty six canine serum samples were collected and screened for VL infection by dual-path platform immunochromatography (DPP®). The positive samples were tested by enzyme-linked immunosorbent assay (ELISA) in order to have each result confirmed. Eight-one samples (12.2%) were found positive by DPP® and about 60% of them were confirmed by ELISA. The high incidence of canine VL associated to the reported presence of the main VL vector, Lutzomyia longipalpis, in Jaboticatubas may favor the transmission cycle of the disease in the municipality.

Financial support: Fundação de Amparo à Pesquisa de Minas Gerais (FAPEMIG), CNPq and FIOCRUZ. - EPIDEMIOLOGY [627] P 765 - CANINE INFECTION AND PHLEBOTOMINE SAND FLY FAUNA IN SABARÁ, AN ENDEMIC MUNICIPALITY IN THE METROPOLITAN AREA OF THE CAPITAL OF THE BRAZILIAN STATE OF MINAS GERAIS. MICHALSKY, E.M.1; LIMA, A.C.1; FRANÇA-SILVA, J.C.2; LARA-SILVA, F.O.1; COSTA, A.A.1; BRITO, J.F.1; SOUZA, M.A.1; DIAS, E.S.1 1.CPQRR, BELO HORIZONTE, MG, BRAZIL; 2.UFMG, BELO HORIZONTE, MG, BRAZIL.

Keyword:canine visceral leishmaniasis; lutzomyia longipalpis; sabará

Abstract: Sabará is a Brazilian municipality with 126,269 inhabitants located in the metropolitan region of Belo Horizonte, the capital of Minas Gerais state. The first cases of canine visceral leishmaniases (CVL) in the whole region were notified in the neighborhood of Sabará. In the present study we aim to verify the current status of CVL as well as the phlebotomine sand fly composition and density in eight districts of that municipality. Two thousand dogs were screened for VL through ELISA and IFA. The prevalence infection rates varied from 2.26% to 5.03%, according to the district, with an average value of 4.25%. Fifty seropositive dogs were necropsied and had tissues (spleen, lymph nodes, skin and bone marrow) collected for further analysis. Bone marrow samples from those dogs were also seeded in culture medium NNN/LIT for parasite isolation. We obtained parasite growth in 38 (76%) of them. Work is in progress to identify the circulating Leishmania species in the municipality. Entomological captures were performed for three consecutive nights per month from January 2011 to December 2012. A total of 482 specimens were captured in the two-year period and the species distribution with respective percentages were as follows: Lutzomyia longipalpis (94.81%), L. intermedia (0.20%), L. sallesi (2.07%) and L. whitmani (1.45%). Although the density of phlebotomine sand flies was apparently low with an average value of 20 specimens per month of capture, L. longipalpis was highly predominant in Sabará. The great majority of specimens (76.55%) were captured in the peridomicile. Our next step will be to determine the natural Leishmania infection rates and the parasite infecting species in those sand flies.

Financial support: FAPEMIG, FIOCRUZ, CNPq. - CONTROL PROGRAMS [631] P 766 - IMPORTANCE OF VECTOR SURVEILLANCE IN THE DEFINITION OF PUBLIC HEALTH ACTIONS IN THE MANAGEMENT OF AN OUTBREAK OF LEISHMANIASIS IN MADRID REGION IRISO, A.1; JIMÉNEZ, M.2; VÁZQUEZ, M.Á.3; GONZÁLEZ, E.2; TELLO, A.3; VILAS, F.1; FÚSTER, F.1; GONZÁLEZ, D.3; MOLINA, R.2 1.SUBDIRECCIÓN GENERAL DE SANIDAD AMBIENTAL, CONSEJERÍA DE SANIDAD, COMUNIDAD DE MADRID, MADRID, SPAIN; 2.UNIDAD DE ENTOMOLOGÍA MÉDICA, SERVICIO DE PARASITOLOGÍA, CENTRO NACIONAL DE MICROBIOLOGÍA, ISCIII, MAJADAHONDA, MADRID, SPAIN; 3.DEPARTAMENTO ZOOLOGÍA Y ANTROPOLOGÍA FÍSICA, FACULTAD CIENCIAS BIOLÓGICAS, UNIVERSIDAD COMPLUTENSE, MADRID, SPAIN.

Keyword:leishmaniasis; control; madrid

Abstract: At the end of 2010 an outbreak of human leishmaniasis caused by Leishmania infantum was detected by the Epidemiological Surveillance System of the Autonomous Government of Madrid in the south of the region. Both epidemiological and environmental measures were taken: the study of the disease in different domestic and wild animals (reservoirs), the collection of sand flies (vector) and the implementation of control measures and communication to the population. The aim of the vector investigation is to determine the abundance and distribution of sand flies in the area affected by the outbreak, to evaluate the level of risk for the population and to guide the fight against the vector and the population at risk. It is a very complex action due to the size of the affected area: a surface of 70 km2 where more than 500.000 people live. The surveillance was developed in 2011 and 2012 with the use of sticky paper and light traps. In the year 2012, twenty-four sampling areas were established with 120 sticky paper traps placed fortnightly, following the results obtained in 2011. In the areas with more presence of the vector, specific samplings were made to identify breeding places and to design actions of habitat destruction and insecticide application in order to reduce sand fly densities. With the aim of determine the rates of infection and the source of the female blood meals 2 CDC light traps were used in 2 locations in September and October. In 2012 were used 4 CDC traps in a parallel study

and these data are presented in another abstract. The study has shown the role of Phlebotomus perniciosus as main vector in the outbreak. It was present in more than 60% of the captures and showed high densities near the location of the human cases. The average density exceeded 50 sand flies/km2, with more than 100 specimens in some locations. The activity of this species extended from May to early October. There were two peaks of activity, one in July and a second one in September. Regarding the vector control measures taken an estimated reduction in the sand fly activity of more than 50% has been achieved in treated locations. A strategy for the 2013 has been established, based on both the destruction of sand fly breeding habitats and the complementary use of insecticides.

- EPIDEMIOLOGY [633] P 767 - COHORT STUDY OF RISK FACTORS FOR THE PERPETUATION OF CANINE VISCERAL LEISHMANIASIS (CVL) IN ENDEMIC AREA IN THE CITY OF BAURU, STATE OF SÃO PAULO, BRAZIL*.

TANIGUCHI, H.H.1; SILVA, S.B.A.1; SANTOS, P.A.1; BARBOSA, J.E.R.1; GONÇALVES NETO, J.R.2; BARBOSA, J.A.R.1; ELIAS, C.R.1; HIRAMOTO, R.M.1; TOLEZANO, J.E.1 1.INSTITUTO ADOLFO LUTZ, SÃO PAULO, SP, BRAZIL; 2.CENTRO DE CONTROLE DE ZOONOSES, BAURU, SP, BRAZIL.

Keyword:canine visceral leishmaniasis; risks factors; reservoirs

Abstract: Visceral leishmaniasis (VL) presents itself in the process of expansion and persistence of foci of transmission in different regions of São Paulo urban areas. The aim of the study was to assess the environmental diagnosis and the importance of canine replacement in the Vila Santa Teresinha, Bauru. Previously this community presented records of dogs naturally infected by Leishmania chagasi. A cohort study was carried out between 2008 and 2012, including 180 households with the presence of dogs. Serological surveys occurred each six months for the diagnosis of canine VL. In the first survey (June 2008 ) of 130 dogs examined 8 were (6.2%) infected. After eight inquiries, 484 dogs examined, 12.0% (58/484) were diagnosed positive, 41 (22.8%) households with the presence of one up to five infected animals. It was found that over 80% of the households with infected dogs replace one or more dogs, and about 22.0% of these households comeback to present infected animals in the following investigations, which confirmed the persistence of the conditions for the maintenance of the foci of transmission. The strategy of performing biannual identification and removal canine reservoirs enabled reduction of the prevalence of canine infection in the first 18 months, followed by a recrudescence in follows surveys. We identified environmental risk factors for colonization of vectors of LVC in households: the existence of terrain or yard vegetation cover, shaded areas; animal feces or decaying organic matter, presence of dogs and other animals. The risks were ranked: absent, mild - up to 3 of these factors, and high - with 4 or more factors. Over 90% of households were classified between medium and high risk environment for canine VL. One should appreciate the quest for environmental sanity, and this should be a priority component in the control of canine VL. *Supported by CNPq – Doenças Negligenciadas.

- DIAGNOSIS – EXPERIMENTAL AND CLINICAL [639] P 768 - DIRECT COSTS OF DIAGNOSTIC TESTS FOR HUMAN VISCERAL LEISHMANIASIS IN BRAZIL MACHADO DE ASSIS, T.S.; GUIMARÃES, P.N.; RABELLO, A.

CENTRO DE PESQUISAS RENÉ RACHOU, FUNDAÇÃO OSWALDO CRUZ, BELO HORIZONTE, MG, BRAZIL.

Keyword:direct costs; diagnostic; visceral leishmaniasis

Abstract: Several diagnostic tests have been used in the management of patients suspected of human visceral leishmaniasis (VL) enrolled in health services in Brazil. Although the performance of these tests is target of several studies, economic aspects are rarely included. The aim of this study was to estimate the direct costs of six diagnostic tests available for VL in Brazil: rapid test IT LEISH® (Bio-Rad Laboratories, USA); rapid test Kala-Azar detect® (INBIOS International, USA); direct agglutination test - DAT-LPC (Laboratório de Pesquisas Clínicas, Centro de Pesquisas René Rachou-CPqRR, Brazil); indirect fluorescence antibody test – IFAT (Instituto de Tecnologia em Imunobiológicos Bio-Manguinhos, Brazil); polymerase chain reaction (PCR) using as target the kDNA of Leishmania and microscopical examination of bone marrow aspirate. In the cost-estimates, only the costs of operational nature were included, assuming that infrastructure and equipment are in place. The main sources of information were the producers of diagnostic kits, the prefecture of Ribeirão das Neves (RN - city of Minas Gerais state, Brazil) and CPqRR. All estimates were calculated taking into consideration the 102 patients with clinical suspicion in RN, during 2011, for whom any diagnostic test was performed by the health system of RN (DAT and rapid tests) or CPqRR (IFAT and PCR).The mean time for test performance - from the collection of biological material until the release of the result - of the rapid tests IT LEISH® and Kala-Azar detect® and the DAT-LPC was of 30, 50 and 40 minutes, respectively, while the mean time for the performance of bone marrow aspirate/examination, IFAT and PCR was of 120, 130 and 190 minutes, respectively. The direct costs of these tests were: DAT-LPC - US$6.9, IT LEISH® - US$7.7, Kala-Azar detect®- US$8.1, IFAT - US$26.7, bone marrow examination - US$27.5, PCR - US$ 43.3. In conclusion, DAT was the diagnostic test with lower direct cost, followed by both rapid tests IT LEISH® and Kala-Azar detect®, while the rapid test IT LEISH® is the faster, followed by DAT and rapid test Kala-Azar detect®. Further studies, evaluating the cost-effectiveness of diagnostic tests for VL are been conducted and may help health managers to set up protocols and diagnostic algorithms.

Funding: Fundacão de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG, Belo Horizonte, Minas Gerais, Brazil; Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz FIOCRUZ, Belo Horizonte, Minas Gerais, Brazil.

- CONTROL PROGRAMS [644] P 769 - ENVIRONMENTAL MANAGEMENT OF AN OUTBREAK OF HUMAN LEISHMANIOSIS IN THE COMMUNITY OF MADRID (SPAIN): PRELIMINARY ACTIONS. MIRÓ, G.1; CARPINTERO, J.2; SEVILLA, S.3; MARTÍNEZ, A.4; FÚSTER, F.5; ORDOBÁS, M.3; DÍAZ, R.2; DE LA FUENTE, S.4; PEROTE, M.2; ARCE, A.3; IRISO, A.5; ESCACENA, C.6; ESTIRADO, A.3; MARINO, E.5; VILAS, F.5 1.DPTO. SANIDAD ANIMAL, FACULTAD DE VETERINARIA (UCM), MADRID, SPAIN; 2.SUBDIRECCIÓN GENERAL DE RECURSOS AGRARIOS, CONSEJERÍA DE SANIDAD, COMUNIDAD DE MADRID, MADRID, SPAIN; 3.SUBDIRECCIÓN GRAL DE PROMOCIÓN DE LA SALUD Y PREVENCIÓN, CONSEJERÍA DE SANIDAD, COMUNIDAD DE MADRID, MADRID, SPAIN; 4.SERVICIO DE SALUD PÚBLICA AREA 9, CONSEJERÍA DE SANIDAD, COMUNIDAD DE MADRID, MADRID, SPAIN; 5.SUDIRECCIÓN GRAL DE SANIDAD AMBIENTAL, DIRECCIÓN GRAL DE ORDENACIÓN E INSPECCIÓN, CONSEJERÍA SANIDAD, MADRID, SPAIN; 6.SUDIRECCIÓN GRAL DE SANIDAD AMBIENTAL, DIRECCIÓN GRAL DE ORDENACIÓN E INSPECCIÓN, CONSEJERÍA SANIDAD, MADRID, SPAIN.

Keyword:leishmania infantum; human leishmaniosis; outbreak

Abstract: Background: Leishmaniosis due to Leishmania infantum, is a zoonotic disease, which is endemic in Spain, and has been notifiable when found in humans in the Community of Madrid since 1997. In the past decade, between 12 and 25 cases per year have been reported in the region. From July 2009 to December 17, 2012 a total of 403 cases have been reported in the southwest area of Madrid with a corresponding incidence rate of 23.21 per 100,000 people. This communication describes the environmental steps taken to control the outbreak.

Methodology: Surveillance activities, research, environmental and epidemiological monitoring coordinated from the Ministries of Health and Environment. The College of Veterinarians of Madrid participated and advice was provided by the Instituto de Salud Carlos III, VISAVET Centre, the Veterinary Faculty and the Faculty of Biological Sciences (Universidad Complutense de Madrid).

Results and Discussion: When the outbreak was detected multiple measures were implemented with the following results: - Reservoir: The low seroprevalence in dogs detected using IFAT (less than 2%) does not explain the human outbreak in this area. An abundant hare population has been detected in the affected area and their possible involvement has been examined. Other reservoirs (rabbits and cats) have also been studied. Xenodiagnosis studies confirmed that the hares were acting as a reservoir (29% of positivity by PCR from skin and spleen biopsies in March 2012), and a Lagomorph Control Plan was launched. A total of 1100 hares and 120 rabbits were captured.

- Vector: Phlebotomus perniciosus was identified in 64% of the over 20,000 specimens collected and high densities of this insect were detected in the outbreak area (45.3 flebotomines/m2 on average). A Vector Control Plan was also launched and included disinsection, cleaning, clearing, etc.

- Environment: Recent environmental changes in the area with the creation of playgrounds, a new road network, etc. have had an impact on the ecology of the sandfly. A plan was set up to establish environmental sanitation measures to eliminate vector habitats, including the cleaning of organic waste, burrows, sewers, etc.

- Communication: creation and distribution of information, participation in talks aimed at professionals and people in the area.

Conclusions -This is the first report of a non-vectorial transmission route for Leishmania infantum infection involving a new reservoir, the lagomorphs, coinciding in an area with a high density of sandflies. Xenodiagnosis test confirms their active role in the transmission cycle.

- EPIDEMIOLOGY [653] P 770 - SOCIO-ECONOMIC FACTORS ASSOCIATED TO THE OCCURRENCE OF CANINE INFECTION BY LEISHMANIA INFANTUM CHAGASI IN TERESINA – PIAUI – BRAZIL MACEDO, E.C.1; NEGREIROS, A.S.1; WERNECK, G.L.2; SILVA, K.M.1; SILVA, K.R.3; CRUZ, M.D.S.P.1 1.UNIVERSIDADE FEDERAL DO PIAUÍ, TERESINA, PI, BRAZIL; 2.UNIVERSIDADE DO ESTADO DO RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 3.CCENTRO DE PESQUISA GONÇALO MONIZ – FIOCRUZ, SALVADOR, BA, BRAZIL.

Keyword:socio-economic factors; canine; leishmaniasis

Abstract: American visceral leishmaniasis (AVL) is a zoonosis caused by protozoan parasites of the Leishmania genus. In Brazil, the disease is caused by Leishmania (infantum) chagasi and the main vector is the phlebotomine sandfly Lutzomyia longipalpis, found both in natural ecotopes and in the rural and urban environments. This vector is known to adapt easily to the peridomestic environment. The domestic dog has been incriminated as the main reservoir of the parasite in the urban environment, but the control measures based on culling seropositive dogs have not shown to be effective to contain the spread of the disease throughout the country. Many studies evaluated risk factors for human visceral leishmaniasis but few focused on the socioeconomic and environmental factors associated with infection among dogs. Knowledge of these factors might help identify the conditions that contribute to the maintenance of transmission cycles in the urban environment and identify new targets for intervention. The objective of this study was to assess the association between socioeconomic and environmental factors and the occurrence of canine leishmaniasis at Teresina city, Brazil. This cross-sectional study was developed in ten districts of Teresina, involving 536 houses and 800 dogs. Peripheral blood samples were collected by vein punction using vacuntainer tubes without anticoagulant for performing serological tests (indirect immunofluorescence test (IFAT) and Enzyme linked immunosorbent assay (ELISA)). Serum samples were considered positive when positive in both tests. Owners of the selected dwellings were interviewed using a semi-structured questionnaire addressing socioeconomic and environment aspects. The association between variables and seropositivity was assessed through multilevel logistic regression models. Global seropositive prevalence was 42%. There was no statistically significant difference between infection prevalence and age and sex of animals, literacy of the household head, presence of other domestic animals or with household characteristic like water supply, inadequate sewage disposal system, type of floor and roof. Non-breed dogs and those living in houses with absence of masory walls, presence of a kennel, and with longer time of dog residency showed higher odds of seropositivity. These results suggest that some peridomestic characteristics, especially the absence of barriers that allow dogs to have a free access to the street, in association to the presence of a kennel might contribute to maintain the infection cycle between dogs. Intervention measures oriented to the management of the peridomestic environment and responsible dog possession could be useful tools for reducing disease burden in endemic area.

- CLINICAL LEISHMANIASIS [654] P 771 - LEISHMANIA INFANTUM CHAGASI AMASTIGOTES IN CEREBROSPINAL FLUID OF A NATURALLY INFECTED DOG CARDINOT, C.B.1; ALMEIDA, B.F.1; LAURENTI, M.D.2; MARCONDES, M.1 1.SCHOOL OF VETERINARY MEDICINE, SÃO PAULO STATE UNIVERSITY, ARACATUBA, SP, BRAZIL; 2.UNIVERSITY OF SÃO PAULO, SÃO PAULO, SP, BRAZIL.

Keyword:zoonotic visceral leishmaniasis; central nervous system; qpcr

Abstract: Visceral leishmaniasis is considered a multisystem disease, and occasionally neurological signs have been observed in naturally infected dogs. There are even few reports of histopathological lesions in central nervous system of asymptomtic dogs, suggesting that parasites or immmunemediate mechanisms are involved in the development of cerebral leishmaniasis.

Although amastigotes have been found in choroid plexus, meninges and central nervous system, only Leishmania sp. DNA has been detected in cerebrospinal fluid. A 3-year-old male mongrel dog was referred to São Paulo State University in Araçatuba, an endemic area for zoonotic visceral leishmaniasis, presenting poor nutritional state, pale mucous membranes, enlargment of palpable lymph nodes, hepatosplenomegaly, generalized desquamative dermatitis, onychopathy, corneal ulcer, joint swellings and lameness. Visceral leishmaniasis was confirmed by cytologic diagnosis and the dog was classified in stage C of the disease. In compliance with a federal law the dog was euthanized. Before euthanasia, under general anesthesia, cerebrospinal fluid (CSF) was collected by cerebellomedullary cisternal puncture. CSF analysis revealed a clear appearance; 13 cell/ l with a predominance of small mononuclear cells (66%), followed by large monunucleares (30%) with some macrophages presenting amastigotes of Leishmania, and intact neutrophils (4%); proteins: 38.7 mg/dL. Although the dog had no neurological signs, because of the presence of amastigotes in CSF, samples from meninges, frontal cortex, thalamus, cerebellum, choroid plexus of lateral ventricles and fourth ventricle were collected and subjected to Real-time PCR for Leishmania DNA amplification. Mean parasite load was 16.964 parasites/mL. In none of the previously reported leishmaniasis neurological cases amastigotes were found in CSF and parasite load in CNS was evaluated. These findings demonstrate that parasites reached central nervous system and even without neurological disorders, If this dog had not been euthanized neurological signs would have probably been developed. Some authors state that the use of anti-Leishmania drugs could led the parasite to escape from the drug in the CNS, however, this dogs had not been treated with any drug.

- DIAGNOSIS – EXPERIMENTAL AND CLINICAL [659] P 772 - THE FORGOTTEN DISEASE OF CATS: A CASE OF FELINE LEISHMANIOSIS BASSO, M.A.1; MARQUES, C.S.1; DUARTE, A.1; PISSARRA, H.1; CARREIRA, L.M.1; VALÉRIOBOLAS, A.2; SANTOS-GOMES, G.2; TAVARES, L.1; FONSECA, I.P.1 1.CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO EM SANIDADE ANIMAL, FACULDADE DE MEDICINA VETERINÁRIA, LISBON, PORTUGAL; 2.CENTRO DE MALÁRIA E OUTRAS DOENÇAS TROPICAIS, INSTITUTO DE HIGIENE E MEDICINA TROPICAL, LISBON, PORTUGAL.

Keyword:leishmaniosis; cat; portugal

Abstract: A 2-years-old, domestic shorthair, male neutered cat, was referred in July 2012 to the Veterinary Teatching Hospital of Veterinary Faculty of Lisbon with fever, multiple nodular lesions located in the ears and head and contaminated ulcers in the tibio-tarsical and carpical regions. Since 2011 the cat has been observed by other veterinarians wich treated the animal for pyodermatitis with prednisolone, amoxycillin/clavulanic acid followed by doxycycline without clinical improvement.

Following the clinical exam, blood was collected for determination of hematological and biochemical parameters, a fine needle aspirative puncture was performed from the skin nodules and the animal was tested for feline immunodeficiency virus specific antibodies (FIV) and feline leukemia virus (FeLV) antigen.

Cutaneous nodular aspirates revealed the presence of Leishmania amastigostes by optical microscopy. To complement this finding, the quantification of antibodies anti-leishmania was

obtained by indirect immunofluorescent assay technique (IFI titre>1280) and qPCR using Leishmania infantum specific probes was performed in blood samples to determine parasitic load. Cultures in Schneider's Insect Medium were also made from the aspirated material, being the isolated promastigotes infective for mouse.

Treatment was iniciated with 10mg/kg allopurinol twice a day. After 15 days without any clinical improvement, treatment was reinforced with 50mg/kg of injectable antimoniate of Nmetilglucamine (Glucantime®), once a day, for 30 days.

The total remission of the skin nodules of the ears and head was achieved with this treatment but, although the healing seemed to be ongoing, the complete closure of the ulcerative lesions located in the hindlimbs was only possible with a surgical approach.

The medical follow-up was done every two days, and serological and molecular testing was performed every two weeks until the end of the injectable treatment. Parasitic load was highly reduced as well as IFI titre (1/320).

The findings indicate that the use of N-metilglucamine antimoniate in associaton with allopurinol in the prescribed dose was safe and clinical cure was achieved. Taking into account that the prevalence of feline leishmaniosis in cats in Portugal has been increasing in the last years it is crucial to consider this differential diagnosis in dermatological lesions.

Funding: Project FCT PTDC/CVT/ 118566/2010; Grant SFRH/BD/778862011 - VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [664] P 773 - GEOMETRIC MORPHOMETRY REVEALS DIFFERENCES BETWEEN NORTHERN POPULATIONS OF NYSSOMYIA NEIVAI FROM ARGENTINA: PRELIMINARY RESULTS UTGÉS, M.E.1; DUJARDIN, J.2; SALOMÓN, O.D.1 1.INSTITUTO NACIONAL DE MEDICINA TROPICAL, PUERTO IGUAZÚ, MISIONES, ARGENTINA; 2.MALADIES INFECTIEUSES ET VECTEURS: ÉCOLOGIE, GÉNÉTIQUE, ÉVOLUTION ET CONTRÔLE (MIVEGEC), IRD, MONTPELLIER, FRANCE.

Keyword:cutaneous leishmaniasis; wing shape variation; ecoregion

Abstract: Nyssomyia neivai is the principal vector of Leishmania sp. causing epidemic Cutaneous Leishmaniasis in nine endemic provinces of Argentina. At the regional scale, studying the heterogenity of populations can give clues to develop common or differential control strategies, to study risk potentiality and migratory routes. If phenotypic variation has environmental and/or genetic causes, its study could help to detect local populations with potentially important characters. Geometric morphometry is a relevant tool for quantifying phenotypic variation and to understand its epidemiological importance.

To assess differences in shape and size between Ny. neivai populations, we compared wing shape of each sex between individuals from two endemic provinces: Jujuy (24 females, 83 males) and Formosa (70 females, 52 males). In each province we sampled 3 and 5 sites respectively. For Jujuy’s females, data from one more site in Jujuy was available. Because of low sample size in some sites, the comparisons were only made between the two regions. We used 10 landmarks on each wing.

We found significant differences in the wing shape of females between regions (p < 0,005).

Sites in Jujuy and Formosa are 400 km apart and belong to different ecoregions (Yungas and Chaco Húmedo, respectively). The discrimination between them was not due to their slight size

difference (size contribution= 3%). Their reclassification based on shape showed 78% of correct validated classification.

No significant differences in wing shape were found for males. When analyzing centroid sizes between sites, we found significant differences (p < 0,005) between sites in Jujuy that were 80 km apart. The greatest variability was found at a site in Formosa that was near a goat pen (CV = 5,6 %); however, sample size at this site was low (n=8).

These are the first results on geometric morphometry of Ny. neivai wings from Argentine populations. Future sampling efforts may help better characterize these sites and other more in other endemic provinces.

- EPIDEMIOLOGY [668] P 774 - OUTBREAK OF AMERICAN CUTANEOUS LEISHMANIASIS (ACL) IN BANDEIRANTES – PARANÁ IN 2007 ROSA CRUZ, M.F.1; ROSA CRUZ, C.F.1; GALATI, E.B.2; MARQUEZ, E.S.1; ROSA CRUZ, M.F.3 1.UNIVERSIDADE ESTADUAL DO NORTE DO PR, BANDEIRANTES, PR, BRAZIL; 2.FACULDADE SAUDE PUBLICA USP SP, SAO PAULO, SP, BRAZIL; 3.UNIVERSIDADE NORTE DO PARANA, BANDEIRANTES, PR, BRAZIL.

Keyword:american cutaneous leishmaniasis; epidemiogical aspects; outbreak

Abstract: Outbreak of American cutaneous leishmaniasis (ACL) in Bandeirantes – Paraná in 2007 Introduction: American cutaneous leishmaniasis (ACL) is a zoonosis that occurs in men and wild or domestic animals. In the state of Paraná, ACL is endemic, out of the 12,304 cases of the disease reported in southern Brazil from 1980 to 2003, 12,220 (99.3%) occurred in the state of Paraná. Objectives: To describe the occurrence of ACL cases in Bandeirantes in 2007, to recognize the socio-economic, cultural and leisure activities, to identify the risk factors, to describe the knowledge of transmission and the means of prevention, transmission and treatment of ACL. Method: A descriptive epidemiological study using data from a questionnaire previously developed and adapted from other epidemiological studies. Results: There were 36 cases of ACL in 2007. Among the 29 individuals interviewed 66% of cases were female and aged over 60 years. There was a predominance of domestic activity (55%) and 25% had completed only the fourth grade of elementary school, 83% had their own house, while 93% are brick with plaster. Most homes have barns, hen houses and pig sties. There are streams in its vicinity and the presence of orchards, 83% of households are connected to sewage networks, 86% have no public service of garbage collection. Regarding frequency in rural areas, 83% of the subjects attend, 41% have relatives and 34% visit relatives at least once a week. Regarding the environment, the majority reported the presence of organic matter and pets. It is observed that the majority of respondents (66%) did not know about ACL and could not inform any form of prevention. Conclusion: The presence of this breakout suggests that there are ecological conditions that would ensure the reproduction and circulation of parasites. The results suggest that transmission of ACL may be occurring in domestic and peridomestic areas. Most of the people interviewed are not aware of the aspects of transmission, symptoms and prevention of ACL, so interventions are needed in the localities, through educational partnerships with health agencies and educational institutions.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [675] P 775 - LUTZOMYIA LONGIPALPIS IS ASSOCIATED TO BOTANICAL FAMILIES IN BRAZILIAN SEMI-ARID REGION SOARES, M.R.A.1; SANTOS, F.F.M.2; LIMA, M.S.C.S.1; COSTA, C.H.N.3 1.CAMPUS AMÍLCAR FERREIRA SOBRAL/ UNIVERSIDADE FEDERAL DO PIAUÍ, FLORIANO, PI, BRAZIL; 2.UNIVERSIDADE DE BRASÍLIA, BRASÍLIA, DF, BRAZIL; 3.INSTITUTO DE DOENÇAS TROPICAIS NATAN PORTELA/UNIVERSIDADE FEDERAL DO PIAUÍ, TERESINA, PI, BRAZIL.

Keyword:leishmaniasis; vector; landscape

Abstract: Background: Lutzomyia longipalpis is the main vector of visceral leishmaniasis (VL) in Brazil with wide distribution. The disease is highly endemic in the semi-arid northeast, where L. longipalpis is found in the wild. Sand flies are associated with plants, which can be a clue for the transmission of VL. Methods: Sand flies were collected in 4 different points of Serra da Capivara National Park, in the semi-arid of southeast Piauí State. A botanic inventory of the area was built by using the line method, with the demarcation of 10 parcels of 10m x 5m. The botanic samples were compared to the herbarium of the Federal University of Piauí. Results: The captures revealed 11 species of the genus Lutzomyia and 31 botanical families. L. longipalpis was the most abundant species (65%), present in all collection points. The botanical families Fabaceae, Convolvulaceae, Poaceae, Malpighiaeceae e Euphorbiaceae were the most abundants. The similarity Sorensen coefficient showed that 3 points were the most similar at the fitossociological point of view. In these areas, the proportion of L. longipalpis fluctuates around 67%. There was a dependency correlation (R2 = 0.89) of the proportion of L. longipalpis to the collection points, when compared to other species of sand flies. Conclusions: This study demonstrated the association of L. longipalpis with botanical families in the semi-arid of Brazil where kala-azar is endemic. The finding indicates the need to map the distribution of L. longipalpis according to the flora, as a possible to tool for the control of the disease.

- EPIDEMIOLOGY [685] P 776 - AMERICAN CUTANEOUS LEISHMANIASIS IN THE STATE OF PARANÁ: A RETROSPECTIVE EPIDEMIOLOGICAL STUDY FROM 2003 TO 2008 BISETTO JR., A.B.1; LUZ, E.2; THOMAZ-SOCCOL, V.3 1.SESA- PR, CURITIBA, PR, BRAZIL; 2.UFPR, CURITIBA, PR, BRAZIL; 3.UNIVERSIDADE POSITIVO, CURITIBA, PR, BRAZIL.

Keyword:american cutaneous leishmaniasis ; retrospective epidemiological study; leishmania (viannia) braziliensis

Abstract: Our group analyzed medical records of clinical cases reported between 2003 and 2008 as part of our ongoing investigation of American cutaneous leishmaniasis (ACL) epidemiology due to Leishmania (Viannia) braziliensis in the State of Paraná. The spatial and temporal distribution of the disease were assessed and the detection coefficient (DC) was determined, according to county and health region. We also correlated the disease with the climatic and geographic data and vector occurrence. Entomological data analyses were complemented by reports of entomological surveillance of ACL in the State of Paraná, conducted by the Entomology Center of the State Health Department (SESA-Secretaria de Estado da Saúde). To more fully understand the epidemiological features of ACL, knowledge concerning the populations at risk is required. Based on SINAM data (a national healthcare database) collected between 2003 and 2008, we verified a total of 3667 records, 64.44% of which involved autochthones of their municipality of residence. The highest number of cases, 959, and consequently the highest detection coefficient (CD), was determined for 2003: 12.02 cases per 100,000 inhabitants. In the remaining years the number of cases was between 460 and 643. Analysis verified that males

aged 10 or over and whose profession is related to agriculture are the individuals most at risk of contracting ACL. The largest number of cases occurred in regions with a Cfa Köppen climate, at altitudes of less than 600m and rainfall between 1200 and 1800mm³. The species Lutzomyia intermedia s.l. is the most abundant (56.7%) and presents the widest distribution among the municipalities and the mesoregion. Lu. whitmani is the second species (31.8%) regarding prevalence and distribution. Lu. migonei is present in all CL transmission foci, but at low frequencies. These three species represent 92.8% of those of epidemiological importance in the State of Paraná. Parasite species were isolated from patient samples and identified by PCR and RAPD and 60 strains belonged to L. (V.) braziliensis. In order to control the disease, sanitary authorities must develop assistance, prevention and sequel rehabilitation activities.

Environmental management and other control actions are only likely to be effective if education and health promotion actions are implemented.

- EPIDEMIOLOGY [686] P 777 - SEROEPIDEMIOLOGICAL STUDIES OF CANINE LEISHMANIOSIS IN ÉVORA (ALENTEJO, PORTUGAL) SCHALLIG, H.D.F.H.1; CARDOSO, L.2; SEMIÃO-SANTOS, S.J.3 1.KONINKLIJK INSTITUUT VOOR DE TROPEN (KIT), ROYAL TROPICAL INSTITUTE, DEPARTMENT OF PARASITOLOGY, AMSTERDAM, NETHERLANDS; 2.DEPARTMENT OF VETERINARY SCIENCES, ECAV, UNIVERSITY OF TRÁS-OS-MONTES E ALTO DOURO (UTAD), VILA REAL, PORTUGAL; 3.CENTER FOR DIAGNOSIS AND RESEARCH IN LEISHMANIOSIS, ICAAM, UNIVERSITY OF ÉVORA, ÉVORA, PORTUGAL.

Keyword:canine leishmaniosis; direct agglutination test (dat); évora, portugal

Abstract: Canine leishmaniosis (CanL) is a systemic chronic condition and severely affected dogs die unless they are adequately treated. Nevertheless, a major proportion of the infected dogs remains apparently healthy and act as carriers of Leishmania, capable of transmitting parasites to the vector, phlebotomine sand flies. Dogs are the main domestic reservoir for human infection where zoonotic leishmaniosis is caused by L. infantum. The present study describes trends in CanL seroprevalence over two decades in the municipality of Évora, covering an area of 1,307.04 km2 in southern Portugal with an average altitude of 300 metres a.s.l. The climate and vegetation are typically Mediterranean, with dry hot summers (32-35°C) and maximum of rainfall in spring and autumn, and mild winters with temperatures rarely going below 5°C. Canine blood samples on filter paper were collected in the years of 1990 (n = 3,614), 1999 (3,563) and 2010 (n = 1,485). The direct agglutination test (DAT) was used for the titration of antibodies specific to Leishmania. The prevalence of DAT seropositive dogs was 3.9% (n = 141), 9.4% (n = 335) and 5.6% (n = 84) in 1990, 1999 and 2010, respectively. The overall seroprevalence was significantly higher in 1999 (9.4%) compared to 1990 (3.9%), but in 2010 a significant decrease in seroprevalence (5.6%) was found compared to 1999. From 1999 to 2010, seroprevalence significantly turned from higher to lower in the contiguous urban areas compared to the contiguous rural ones. Out of the 141 seropositive dogs in 1990, 113 (80.1%) were apparently healthy and 28 (19.9%) were considered clinically suspect (0.8% of the total study population in 1990). Similar observations were made in 1999 and 2010, where 263 healthy dogs (78.5%) and 72 (21.5%) suspect dogs (2.02% of the total study population) were DAT positive and 71 (80.1%) healthy and 13 (9.9%) suspect dogs (0.87% of the total study population) were seropositive, respectively. Lymphadenopathy, followed by onychogriposis and skin involvement, were the most frequently observed clinical signs. A significant number of dogs of Évora are seropositive for leishmaniosis. A peak in seropositivity was observed in 1999 and there was a decline in the number of seropositive cases in 2010, to the levels of 1990. There was a considerably high proportion of dogs that are seropositive but appear as clinically healthy.

Measures to control human zoonotic leishmaniosis should remain focussed on the canine population.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [689] P 778 - COMPARISON OF TWO TYPES OF LIGHT TRAPS TO CAPTURE SAND FLIES: WHITE AND BLACK LED LIGHT. FERNANDEZ, M.S.1; SANTINI, M.S.2; GOULD, I.T.2; MARTINEZ, M.F.3; BERROSPE, P.E.2; MANTECA-ACOSTA, M.4; PEREZ, A.A.5; SALOMON, O.D.3

1.CENTRO NACIONAL DE DIAGNÓSTICO E INVESTIGACIÓN EN ENDEMO-EPIDEMIAS / CONICET, BUENOS AIRES, ARGENTINA; 2.CENTRO NACIONAL DE DIAGNÓSTICO E INVESTIGACIÓN EN ENDEMO-EPIDEMIAS, BUENOS AIRES, ARGENTINA; 3.INSTITUTO NACIONAL DE MEDICINA TROPICAL / CONICET, PUERTO IGUAZÚ, ARGENTINA; 4.INSTITUTO NACIONAL DE MEDICINA TROPICAL, PUERTO IGUAZÚ, ARGENTINA; 5.DPTO DE ECOLOGÍA, GENÉTICA Y EVOLUCIÓN, FACULTAD DE CIENCIAS EXACTAS Y NATURALES, UBA, BUENOS AIRES, ARGENTINA.

Keyword:sandflies; traps; leds

Abstract: In Argentina leishmaniasis is an endemic and epidemic disease that it has expanded its distribution in recent years. To study the presence and abundance of sandflies involved in the transmission of this disease, we have developed an economic and new type of light traps using white-light LEDs (LLW) and black-light LEDs (LLB). The new traps are a modified CDC minilight trap with a leds ring. The aim of this work was to compare the LLW and LLB traps in relation to phlebotominae captures. A total of 542 traps were located in different sites: peridomiciles of households in urban and rural areas and forest environments, placed in four provinces of Argentina. One LLB and one LLW traps were placed in each site overnight (placed on consecutive nights, randomized one trap per night). Only 76 sites resulted positives for phlebotominae captures (41/190 urban, 24/55 rural and 11/26 forest), with a total of 2711 sandflies captured. Nine species where present: Lutzomyia longipalpis (50.8%), Nyssomyia whitmani (29.4%), Migonemyia migonei (17.3%) and the remaining species representing 2.5% of the total captures (Nyssomyia neivai, Psathyromyia shannoni, Evandromyia cortelezzii, Ev. sallesi, Micropygomyia quinquefer, Pintomyia pessoai and Brumptomyia sp.). Positive association was found between the estimated abundance with both wavelengths for both sexes (rs=0.83, p<0.0001 for females and rs= 0.32, p=0.005 for males) but the LLB trap captured more specimens than the LLW trap (LLB median abundance = 2.0 (IQR=1;7) and LLW median abundance 1.0 (IQR=0;2) for females; LLB median abundance = 5.5 (IQR=1;24) and LLW median abundance 1.0 (IQR=0;3) for males). When an analysis comparing sex abundances was performed for the three main species we found that this pattern of more captures in LLB trap in relation to LLW trap was conserved for Lu. longipalpis in both sexes and for Ny. whitmani only for females. We didn't found differences in the median abundance for Mg. migonei for any sexes and for Ny. whitmani for males. These results suggest the importance of using the same type of trap to compare abundances to avoid biases (in species capture and sex ratio) due to the methodology. Based on this preliminary analysis we suggest using LLB instead of LLW in the studied region for this type of light traps.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [690] P 779 - SEASONAL POPULATION DYNAMICS OF LUTZOMYIA LONGIPALPIS IN CORRIENTES, ARGENTINA MIÉREZ, M.L.; REA, M.J.; BORDA, C.E.; MOSQUEDA, L.A. CENTRO NACIONAL DE PARASITOLOGÍA Y ENFERMEDADES TROPICALES (CENPETROP), FACULTAD DE MEDICINA, UNNE, CORRIENTES, ARGENTINA.

Keyword:seasonal population ; lutzomyia longipalpis ; corrientes, argentina

Abstract:

The Province of Corrientes is located in the subtropical area of the Northeast region of Argentina. The presence of Lutzomyia longipalpis was evidenced since 2009 in the urban area of Corrientes city. Entomological surveillance actions were triggered initially aimed to determine the distribution of the vector, Lu. longipalpis through entomological survey in an endemic zone of canine visceral leishmaniasis in the city of Corrientes (27°28 00 S, 58°50 00 W).

This study examined the seasonal fluctuations of population densities of Lu. longipalpis in the urban zone of Corrientes.

The prevailing climate is subtropical without dry season, with abundant rainfall and high temperatures with little daily or seasonal variations. The average annual temperature for this city is 21°C and its mean annual rainfall, 1,650 mm.

Sand flies were collected weekly from April 2010 to December 2012. CDC light traps were installed at dusk and were operated about 12 hours, for three consecutive nights, in the peridomicile of a house. Traps were placed in two different habitats: hen-house and rabbit burrow with pigeon. Basic environmental conditions, including temperature and relative humidity, were recorded daily.

A total of 12,119 sand flies were trapped. Sand fly numbers captured over the years 2010, 2011 and 2012 were 2,780, 5,358 and 3,981, respectively.

Collection was highest in the rabbit burrow with pigeon (83.5%) compared to hen-house (16.5%).

Sand flies were present during most of the year. Males had the highest number (80%).

In 2010 capture numbers showed peaks in April, May, October, November and December.

During 2011 and 2012, peaks in abundance were observed from January to May. For the three years, temperatures ranged between 16-28°C and humidity from 64-85%.

The highest sand fly collection in one night was collected in April 2010, 385 specimens (81% males, 19% females) at 22°C and 78 % humidity. Sand fly numbers decreased in June to July as the climate became cooler. No sand flies were trapped during the cooler months of July 2010 and 2011. But in winter 2012, five males were collected at 9°C and one at 4ºC during June and seven males at 13°C in July.

The warmest temperatures recorded in April 2012, reaching 39°C (83% humidity) in three serial nights. In these nights 426 sand flies (79% males and 21% females) were collected.

The relationship between the seasonal population dynamics of Lu. longipalpis and abiotic factors will contribute to a broader integrated sand fly control program in Corrientes. For this reason, control strategies based on reducing sand fly populations and health education of inhabitants about the prevention ways from transmission are necessary.

Financial support by SGCYT- Salud Investiga Ramón Carrillo

- EPIDEMIOLOGY [691] P 780 - BATS: RESERVOIRS OR ACCIDENTAL HOSTS OF LEISHMANIA SPP.?

NUNES, C.M.1; OLIVEIRA, F.M.1; BARROS, T.L.1; CARVALHO, C.1; COSTA, L.H.C.2; PEDRO, W.A.1; QUEIROZ, L.H.1

1.UNESP, ARAÇATUBA, SP, BRAZIL; 2.UNIP, ARAÇATUBA, SP, BRAZIL.

Keyword:leishmaniasis; reservoirs; bats

Abstract: Visceral leishmaniasis is widespread in Brazilian urban areas nowadays and some animal species are known reservoirs of this zoonosis, specially the dog. This study aimed at investigating the presence of Leishmania spp. DNA in bats in order to check if they play a role in the transmission. Leishmania (L.) amazonensis DNA has already been detected in bats near São Paulo city, a non-endemic visceral leishmaniasis (VL) area and also Leishmania (L.) infantum chagasi DNA has been amplified in one bat in the Venezuela, in the preserved area and non-endemic for VL. Spleen and skin samples were collected from 375 bats from 19 cities of the São Paulo State, Brazil, which are located in an area of intense transmission of visceral leishmaniasis. Leishmania ssp. kDNA amplification by PCR was positive in 4.5% of the bats; higher positivity was seen in spleen samples (94.1%) and two bats were positive for both skin and spleen samples. Among positive samples, 14 were differentiated by RFLP-PCR of the internal transcribed spacer region (ITS-1) with HaeIII enzyme and 14.3% of the samples showed identical Leishmania (L.) infantum chagasi pattern whereas 64.3% showed restriction pattern of Leishmania (L.) amazonensis. These results demonstrate the occurrence of different species of Leishmania in bats from an endemic area for visceral leishmaniasis, confirming them as accidental hosts. This also alerts to the possibility of bats being potential reservoirs for leishmaniasis and thus contributing to transmission of this zoonosis.

- RESERVOIRS [692] P 781 - BATS AS RESERVOIRS OF LEISHMANIASIS HERNANDEZ, C.G.1; BENTO, E.C.2; BATISTA, L.R.1; TIBURCIO, M.G.S.1; SILVA, E.L.1; RAMIREZ, L.E.1 1.UNIVERSIDADE FEDERAL DO TRIÂNGULO MINEIRO, UBERABA, MG, BRAZIL; 2.INSTITUTO FEDERAL DO TOCANTINS, PALMAS, TO, BRAZIL.

Keyword:bat; leishmania; reservoirs

Abstract: The species of the genus Leishmania transmitted by insects subfamily Phlebotominae are able to infect hosts that vary according to the ecotope. Were described more than 100 species of mammals as potential reservoirs of Leishmania, however was not elucidated the involvement of individuals from order Chiroptera in Leishmania´s life cycle, these mammals live in places suitable for the presence of phlebotomines and therefore at risk acquiring the parasite making them a good reservoir. Due to the lack of information about the participation of bats in leshimaniasis cycle, the goal of this study was the diagnosis of leishmaniasis in bats captured in the Triângulo Mineiro and Alto Paranaiba. Methods: PCR was performed with blood by 93 bats belonging to species captured in the Triângulo Mineiro and Alto Paranaiba by mist nets. Were used for PCR primers JW11 and JW12 specific to the genus Leishmania targeting a conserved region of kDNA that when a product generates positive 120pb. The PCR products were visualized on 6% acrylamide gel and stained with silver. Results: 93 blood samples from bats was analyzed, 8 (8.6%) were positive by PCR for leishmaniasis: 3/8 (37.5%) Glossophaga soricina; 4/8 (50%) Molossus molossus and 1/8 (12.5%) Phyllostomus hastatus. Discussion: Has been demonstrated indirectly leishmaniasis bats in the Old World, through the detection of antiLeishmania (Mutinga, 1975; Morsy, 1987). There is only one report of infection with Leishmania chagasi just a bat (Carollia perspicillata) in Venezuela (Lima, 2008). This report and our finding indicate that bats may be playing an important role in maintaining the cycle of

leishmaniasis in the link between domestic and sylvatic cycle. For identification of Leishmania’s species the samples will be sequencing.

- CONTROL PROGRAMS [707] P 782 - PROGRAM FOR CONTROL OF LEISHMANIASIS: A LOOK AT THE MUNICIPALITY OF CARUARU, PERNAMBUCO STATE COSTA, P.L.1; DE FRANÇA, S.H.F.2; AGRA, M.C.R.2; DE SOUZA, C.L.2; DE SOUZA, C.A.C.2; DA SILVA, E.O.2; DE MELO, S.D.2 1.CENTRO DE PESQUISAS AGGEU MAGALHÃES, RECIFE, PE, BRAZIL; 2.SECRETARIA MUNICIPAL DE SAÚDE, CARUARU, PE, BRAZIL.

Keyword:visceral leishmaniasis; dogs; entomological survaillence

Abstract: Visceral leishmaniasis (VL) is a zoonotic disease caused by Leishmania infantum chagasi (Kinetoplastida: Trypanosomatidae), a parasite transmitted to humans through the bite of infected females of Lutzomyia longipalpis (Diptera: Psychodidae), which has as a reservoir, the dogs, it may submit asymptomatic infection or oligosymptomatic. Rural areas of the municipality of Caruaru, Pernambuco, Brazil, are monitored annually before the constant change in the epidemiological profile of leishmaniasis and the modulation of population density of sandflies. Between the period 2010 to 2012, a total of 456 (44 %) localities was researched for leishmaniasis, being collected 3,223 blood samples from dogs. All samples were sent to the Laboratory of Endemics of IV Regional Health. 1,050 (32 %) samples were positive for the detection of L. infantum chagasi. Even as a component of the program, there were 5,765 pulverization (71 %), according to the agreement on the Programming of the Shares of Health Surveillance (PAVS), Ministry of Health. However, it was observed that the entomological surveillance that is one of the most important components of the program is still very early in the city. Considering the above, and notorious ensure the sustainability of the actions of the program of control of leishmaniasis, seen the importance of knowing the epidemiology of the disease in the municipality of Caruaru, in addition to make the component of entomological surveillance a practice routine, thereby to understand better the behavior vectorial with purpose of guiding the development of improved control strategies.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [717] P 783 - LIFE CYCLE AND CHARACTERIZATION OF IMMATURE FORMS OF LUTZOMYIA WHITMANI CREATED UNDER EXPERIMENTAL CONDITIONS IN SÃO LUÍS - MA DUAILIBE, R.F.1; MELO, M.N.2; REBELO, J.M.M.1; FILHO, A.A.P.2; MORAES, J.L.P.3; SILVA, S.O.2 1.UFMA, SAO LUIS, MA, BRAZIL; 2.UFMG, BELO HORIZONTE, MG, BRAZIL; 3.FUNASA, SÃO LUIS, MA, BRAZIL.

Keyword:tegumentar leishmaniasis ; l. whitmani ; cycle of life

Abstract: This study determined the cycle of life and characterized the immature forms of Lutzomyia whitmani incriminated to be the vector of the Tegumentar Leishmaniasis (LT) in Maranhão. In order to, establish and maintain a colony of L. whitmani in the laboratory, individuals were collected in the municipal of Axixá, endemic area of Leishmaniasis in Maranhão. It was found that the total life cycle was completed after 39,3 days, eggs revealed an ellipsoid shape, whereas the larvae’s exhibited a translucent to yellow cuticles and the pupas had a constant size of 1.5 mm. The hatching offspring represented a sexual relation of 1 male to 1.3 females. Temperature and humidity parameters of colony that minimize a contamination with fungal and mite varying

between 26.20C to 270C and 65% to 72%. In summary, the life cycle of L. whitmani is about a month and ten days since maintained the standards of hygiene and control of physical parameters established in the colony. These data are important for understanding the biology of the vector, as well as for the optimization of techniques for the maintenance of colony, useful to support research in other areas and develop strategies for control and prevention of LT. - VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [721] P 784 - ANALYSIS OF VECTORIAL CAPACITY OF LUTZOMYIA WHITMANI TO LEISHMANIA AMAZONENSIS AND LEISHMANIA BRASILIENSIS IN EXPERIMENTAL CONDITIONS DUAILIBE, R.F.1; REBELO, J.M.M.1; MELO, M.N.2; FILHO, A.A.P.2; MORAES, J.L.P.3; SILVA, S.O.2 1.UFMA, SÃO LUIS, MA, BRAZIL; 2.UFMG, BELO HORIZONTE, MG, BRAZIL; 3.FUNASA, SÃO LUIS, MA, BRAZIL.

Keyword:l. whitmani ; vectorial capacity ; diffuse cutaneous leishmaniasis

Abstract: In this study we examined, whether L. whitmani is susceptible to experimental infection by L. brasiliensis, causing Cutaneous and Cutaneous - Unfledged Leishmaniasis, and L. amazonensis, causing Diffuse Cutaneous Leishmaniasis. L. whitmani individuals were collected to establish a colony free from these parasites. Mice were experimentally infected with L. brasiliensis or L. amazonensis parasites. Female of this L. whitmani colony were permitted to feed on mice exhibiting clinical signs of the disease. Phlebotominae infected with parasites were detected by amplifying a Leishmania 120 bp specific kinetoblast DNA by means of PCR. The L. whitmani infection rate for Leishmania brasiliensis and Leishmania amazonensis was 65.2% and 47.4%, respectively and confirmed that the vector is susceptible to experimental L. brasiliensis and L. amazonensis infection. Comparison of the infection rates revealed statistically significant differences. The high L. whitmani infection rate together with epidemiological and entomological studies, verified that L. whitmani is responsible for L. brasiliensis infection.

Remarkably, here we report for the first time that L. whitmani is vulnerable to experimental L. amazonensis infection and may be a vector of this pathogen and the distributor for Diffuse Cutaneous Leishmaniasis.

- CLINICAL LEISHMANIASIS [726] P 785 - MODE OF LEISHMANIA (VIANNIA) BRAZILIENSIS ENDEMICITY MAINTENANCE IN CORTE DE PEDRA – BAHIA, BRAZIL SILVA, J.1; GIUDICE, Â.1; QUEIROZ, A.1; GUIMARÃES, L.H.1; MACHADO, P.R.L.1; SALGADO, K.1; LAGO, E.1; WILSON, M.E.2; CARVALHO, E.M.1; SCHRIEFER, A.1 1.UNIVERSIDADE FEDERAL DA BAHIA, SALVADOR, BA, BRAZIL; 2.UNIVERSITY OF IOWA, IOWA CITY, UNITED STATES.

Keyword:leishmania (viannia) braziliensis; outbreaks; epidemiology

Abstract: American tegumentary leishmaniasis (ATL) may result from human infection with Leishmania (Viannia) braziliensis, which causes three forms of disease: cutaneous (CL), mucosal (ML) and disseminated leishmaniasis (DL). CL, ML and DL can be found in Corte de Pedra (CP), Bahia / Brazil. In the present study we aim to progressively clarify the mechanism of maintenance of the ATL endemics in CP. We assessed whether the human cases of ATL in that region occur in clusters indicative of outbreaks or isolated like sporadic cases. Two-hundred twenty five CL, 35 ML and 76 DL cases (336 ATL) were enrolled in CP between 2008 and 2011. Geographic coordinates of their living sites were collected by GPS and plotted onto a high definition, geo-

referenced, satellite photograph of the region, using a geographic information system (GIS).

Each form of ATL was analyzed separately. For each new diagnosed case of that particular form of ATL (i.e. CL, DL or ML), the distances for the preceding cases that occurred in the 3, 6 and 12 preceding months were calculated using the GIS tool. Then the correlation between the frequencies of recent cases closest to newly diagnosed cases and incremental distances to these newly diagnosed cases were evaluated. Overall, CL and DL showed significant inverse correlations between frequencies of recent cases and distances to newly diagnosed cases in most time strata taken into account, suggesting that outbreaks are more frequent than sporadic cases of these diseases in CP. ML did not display correlations suggestive of clusters of disease. This suggests that ML may occur as sporadic cases, although a larger sample is necessary for reinforcing this conclusion. We also sought to shed some light on the clonality of these infections within the host. In a preliminary evaluation, parasites of 94 of the ATL patients enrolled in this study were cultured and genotyped by sequencing of a polymorphic locus starting at position 3074 in chromosome 24 of the parasites of CP. Eighty seven percent of these isolates showed only 1 or 2 haplotypes of SNPs and indels detectable for the CHR24/3074 locus, suggesting that these infections presented one or two predominant clones of L. (V.) braziliensis. However this analysis needs to be expanded to biopsy specimens, which better reflect what occurs in the host tissues, particularly given the tendency of some clones of the parasite to adapt better to in vitro culturing than others. All in all, the picture that is emerging is of oligoclonal human infections, which occur in clusters for CL and DL, and as sporadic cases of ML. - EPIDEMIOLOGY [732] P 786 - MOLECULAR SURVEY OF LEISHMANIA, EHRLICHIA/ANAPLASMA, RICKETTSIA AND BABESIA/THEILERIA GENUS IN DOGS FROM MAIO ISLAND, CAPE VERDE MARQUES, C.S.1; SILVA, D.1; DUARTE, A.1; FONSECA, I.P.1; MALTA, M.2; VILAÇA, R.2; PEREIRA, C.2; ALMEIDA, C.2; VIEIRA, S.3; TAVARES, L.1

1.CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO EM SANIDADE ANIMAL, FACULDADE DE MEDICINA VETERINÁRIA, LISBOA, PORTUGAL; 2.VETERINÁRIOS SEM FRONTEIRAS DE PORTUGAL, FACULDADE DE MEDICINA VETERINÁRIA, LISBOA, PORTUGAL; 3.DELEGAÇÃO DO MINISTÉRIO DO DESENVOLVIMENTO RURAL, ILHA DE MAIO, CAPE VERDE.

Keyword:survey; leishmania; cape verde

Abstract: Maio Island is one of the 10 islands from the archipelago of the Republic of Cape Verde located in the Atlantic Ocean, 500 km from West Africa coast.

Canine vector borne diseases (CVBD), including Leishmaniosis, are globally distributed and are spreading rapidly. Information regarding CVBD distribution is therefore importante not only for the local population of dogs and humans but also for travelling people and their pets.

This study aimed to contribute to a better knowledge of the presence of hemoparasites such as Leishmania, Anaplasma/Ehrlichia, Rickettsia and Babesia/Theileria genus in stray dogs from Maio Island, Cape Verde.

In June of 2011 total blood samples were collected from 77 dogs attended under the scope of one Veterinarians Without Frontiers mission. The DNA extraction was performed with the commercial kit DNeasy ® Blood & Tissue Kit (Qiagen, Germany) and detection of Leishmania, Ehrlichia/Anaplasma spp., Rickettsia spp. and Babesia/Theileria spp. DNA was done by qPCR using Taqman® probes.

In this study, Leishmania DNA was not detected in blood samples of any of the 77 dogs included. Ehrlichia/Anaplasma was the most frequent CVBD, present in 70.13% (N=54) dogs, followed by Babesia/Theileria (3.90%, N=3) and Rickettsia (1.30%, N=1). Considering all the CVBDs tested, 3.90% (N=3) of dogs were co-infected with more than one agent and only 28.57% (N=22) were negative for all of them.

To our best knowledge, no study has yet found Leishmania or their vectors, phlebotomine sand flies, in Cape Verde archipelago. This study on Maio Island, also did not find any dog infected with this protozoan. Considering the high number of travelers and their pets, from Leishmania endemic countries, that visit the Republic of Cape Verde, periodic surveys of dogs and vectors, should be implemented to assure rapid epidemiological data flow to local medical doctors and veterinarians.

Besides Leishmania our results revealed a high frequency of infected dogs with other CVBD, especially Ehrlichia/Anaplasma and to a lesser degree Babeisa/Theileria. Further studies are needed to determine the pathogenic and zoonotic degree of these hemoparasites infections found in Maio Island.

Funding: This study was funded by the Portuguese Foundation for Science and Technology and the European Union (FEDER) through Project PTDC/CVT/118566/2010; PhD Grant SFRH/BD/77886/2011 - EPIDEMIOLOGY [734] P 787 - PREVALENCE OF AMERICAN CUTANEOUS LEISHMANIASIS IN SCHOOL CHILDREN OF JATAIZINHO, PARANÁ, BRAZIL CALDART, E.T.; ROCHA, K.F.; CEZAR, T.L.; EGEA, A.C.; FREIRE, R.L.; NAVARRO, I.T.; LOPES-MORI, F.M.R.; MITSUKA-BREGANÓ, R. UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL.

Keyword:seroprevalence; leishmaniasis; children

Abstract: The multiplicity of factors involved in the transmission of the American cutaneous leishmaniasis (ACL) constitutes a challenge to its control. The knowledge of such factors may contribute to redefine the control strategies. The aim of this study was to determine the prevalence of anti-L. amazonensis IgG antibodies and associated factors to infection in 236 school children from one to 15 years old. A questionnaire was applied and analyzed with EpiInfo (3.4.3) statistics software. Serum samples were collected, from August 2011 to October 2012, submitted to Indirect Fluorescence Antibody Test (IFAT) and titers 40 were considered positive. The sample was consisted of: 53.1% female gender; 99.6% from urban area and 50.7% with per capita income between two and four times the minimum wage. Antibodies to L. amazonensis were found in 14 of 236 (5.9%) children distributed as: 7 of them showed a titer of 40; four of 80 and three of 160. Data analysis revealed no association of children leishmaniasis and the variables described. An assay performed in endemic area from Northern Paraná in 1996 found 8.5% (58/683) of positive humans, with varying ages. Another one realized in 2012 in Jataizinho showed a seroprevalence of 5.5% (4/73) just in adulthood people. The prevalence found in this study agrees with the studies previously cited. Moreover, suggests that the number of seroprevalent people has not increased and that there is no difference in prevalence between the various age groups.

- EPIDEMIOLOGY [741] P 788 - ENTOMOLOGICAL ASPECTS RELATED TO THE TRANSMISSION OF VISCERAL LEISHMANIASIS IN BELO HORIZONTE, MINAS GERAIS, BRAZIL SILVA, F.O.L.E.1; SILVA, M.A.1; MICHALSKY, É.M.1; FERREIRA, E.C.1; SCOFIELD, K.M.1; FERREIRA, J.B.1; COSTA, A.J.1; PAIXÃO, F.R.1; SAID, R.F.2; XAVIER, I.M.2; NASCIMENTO, J.C.2; VIEGAS, R.P.2; FIÚZA, V.O.2; DIAS, E.S.1 1.CENTRO DE PESQUISAS RENÉ RACHOU/FIOCRUZ, BELO HORIZONTE, MG, BRAZIL; 2.SECRETARIA MUNICIPAL DE SAÚDE, BELO HORIZONTE, MG, BRAZIL.

Keyword:visceral leishmaniasis; belo horizonte; sand flies

Abstract: Since 1993, human and canine cases of visceral leishmaniasis (VL) have been reported in Belo Horizonte, the capital of the Brazilian state of Minas Gerais. In 2010, 134 human cases were reported to the public health services and the disease was considered a serious health problem with high incidence and mortality. The objectives of this study were to survey the diversity, population, blood feeding sources and natural rates of infection of phlebotomine sand flies captured in two districts of Belo Horizonte, named Miramar and Salgado Filho. Entomological captures were performed with HP light traps from September of 2010 to October of 2012, in nine spots per district. The traps were mounted from 18:00 to 8:00 during three consecutive nights per month. The captured males and females were used for species and blood feeding source identification, respectively. A total of 5,099 specimens were captured with a high predominance of Lutzomyia longipalpis (96.7% of the total of specimens) mostly in the peridomiciles (88.8%). Other species captured with their relative percentages were: Lutzomyia intermedia (0.12%), L. ischyracantha (0.04%), L. whitmani (0.10%), complex cortelezzii (1.86%) and Lutzomyia sp. (1.16%). Five out of 27 engorged sand fly females analyzed displayed the 359 bp fragment of the cytochrome b gene of vertebrates. DNA sequencing of those fragments is in progress aiming the vertebrate identification. Nested PCR (LnPCR) of total phlebotomine sand fly DNA with adequate primers for the Leishmania specific SSUrRNA gene, showed the expected 353bp fragment in 23 samples of L. longipalpis and one pooled sample of cortelezzi complex, among 47 samples tested. Further DNA sequencing of those fragments will allow the identification of the infecting Leishmania at specific level. Minimum infection rates were calculated as 33.8%, and 14.3% for L. longipalpis and the cortelezzi complex, respectively.

The high infection rate of L. longipalpis by Leishmania spp. is in accordance with the high prevalence of canine and human cases of VL in Belo Horizonte. We hope our data will contribute to a better understanding of the epidemiology of VL transmission in Belo Horizonte.

Financial Support: FAPEMIG, FIOCRUZ, Secretaria Municipal de Saúde de Belo Horizonte, CNPq.

- RESERVOIRS [747] P 789 - INFECTIVITY STUDY OF THREE RODENTS HOSTS OF LEISHMANIA (VIANNIA) BRAZILIENSIS: POSITIVITY, PARASITE LOAD AND TISSUE TROPISM ZAMPIERI, R.A.1; JORGE, C.L.1; DA SILVA, M.F.L.1; ANDRADE, M.S.2; BRANDÃO-FILHO, S.P.2; WINTER, L.M.F.1; SHAW, J.J.3 1.BIOSCIENCES INSTITUTE - UNIVERSITY OF SAO PAULO, SAO PAULO, SP, BRAZIL; 2.FIOCRUZ, RECIFE, PE, BRAZIL; 3.BIOMEDICAL SCIENCES INSTITUTE - UNIVERSITY OF SAO PAULO, SAO PAULO, SP, BRAZIL.

Keyword:reservoirs; real time pcr; ssu rdna

Abstract: For over three decades, the primary reservoirs of Leishmania (Viannia) braziliensis were not known, but the development of PCR protocols to detect parasites in samples obtained from sylvatic animals implicated some vertebrate species such as rodents and marsupials as primary reservoirs of L. (V.) braziliensis. To observe the infection profile in some of these reservoirs, animals from established colonies of Nectomys squamipes, Bolomys lasiurus and Rattus rattus were experimentally infected with 106 promastigote forms of L. (V.) braziliensis. After six weeks of infection, samples of skin, spleen and liver were processed to purify template DNA for detection and quantification PCR assays, using DNeasy Blood & Tissue kit (Qiagen), according to manufacturerâ&#x20AC;&#x2122;s protocol. The initial detection protocol consisted in a nested PCR assay using two pairs of SSU rDNA (Small Subunit Ribosomal gene) derived oligonucleotides. The first PCR used primers that amplify a conserved region of all trypanosomatids and the second reaction used primers that amplify a common region of the Leishmania genus. The quantification protocol consisted in a real time SYBR-Green PCR, wherein the parasite load was estimated by normalizing the number of SSU rDNA copies per host glyceraldehyde-3-phosphate dehydrogenase (gapdh) copy number. We observed a higher positivity in samples of B. lasiurus and N. squamipes indicating that these species are more susceptible than R. rattus. However, all samples presented a low parasite load, 11.4, 128 and 235 SSU rDNA copies in R. rattus, N. squamipes and B. lasiurus samples, respectively, what correspond to the DNA of at most one parasite/50 ng of host DNA. Besides, we were not able to observe parasite tissue tropism for the three analysed species. The high positivity in samples of B. lasiurus and N. squamipes, and concomitant low parasite load, may be features that support the life cycle of the parasite, ensuring the role of these rodents as reservoirs in sylvatic cycle.

- EPIDEMIOLOGY [752] P 790 - LEISHMANIA TROPICA LIFE CYCLES IN THE CUTANEOUS LEISHMANIASIS FOCUS OF TATAOUINE REGION, SOUTHEASTERN TUNISIA BOURATBINE, A.1; BOUSSLIMI, N.1; TABBABI, A.2; RAOUANE, M.3; AOUN, K.1 1.LR 11-IPT-06 INSTITUT PASTEUR DE TUNIS, TUNIS, TUNISIA; 2.LR 11-IPT-06, INSTITUT PASTEUR DE TUNIS, TUNIS, TUNISIA; 3.REGIONAL DIRECTORY OF HEALTH, TATAOUINE, TUNISIA.

Keyword:leishmania tropica; cutaneous leishmaniasis; phlebotomus sergenti, north african gundi

Abstract: Leishmania tropica zymodeme MON-8 (syn L. killicki ) is a causative agent of cutaneous leishmaniasis (CL) in Tataouine region, Southeastern Tunisia, North Africa. This area is situated in the northeastern part of the Sahara Desert; and is located in a mountainous area under arid climate. An epidemiological survey recording all CL cases due to this Leishmania species in the region showed that cases were geographically scattered, which suggests a zoonotic reservoir host. The purpose of our work was to explore L. tropica transmission cycles in this region. Sand flies specimens were collected by CDC light traps in and around houses of L. tropica CL cases and in the habitats of the North African gundi which is extremely abundant in natural and peridomestic environments of the area and highly suspected as reservoir. Sand flies were identified according to morphological characters and by a comparative sequence analysis of cytochrome b gene to distinguish between Phlebotomus (P.) chabaudi and P. riouxi. Live female specimens were dissected; the head and genitalia were used for species identification; The gut was examined for promastigotes detection. In case of positivity, cultures were performed in NovyNicolle-MacNeal medium. Gundis were manually caught by the local population near the entrances of caves and crevices inhabited by the rodent and then kept alive in cages; Animals were physically examined; Blood samples and nasal biopsies were used for DNA extraction.Leishmania infection was evaluated by real-time kinetoplast DNA polymerase chain

reaction. Species identification was performed by internal transcribed spacer one (ITS1)-PCRrestriction fragment length polymorphism and high-resolution melting analysis of the 7SL RNA gene.Diversity analysis showed that P. sergenti was the most abundant sandfly species inside houses whereas P. riouxi was the most abundant in the natural habitat of gundis. Both P. sergenti and P. riouxi were equally present around houses in anthropogenic and semianthropogenic habitats. This distribution was consistent with the possibility of two transmission cycles: P. riouxi transmitting L. tropica among wild populations of rodent and P. sergenti transmitting the same parasite strains to humans within or near their houses. L. tropica was identified after a dissection and culture in Phlebotomus sergenti caught inside the house of CL patient and by a real-time PCR in five gundis. Alignments of the ITS-1 DNA sequences and 7SHRM curves analysis indicated that similar genotypes were present in humans, a sandfly and gundis from the same region. This suggests that P. sergenti and the North African gundi are respectively, the vector and the reservoir host of L. tropica in Southeastern Tunisia.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [755] P 791 - QUANTIFICATION OF LEISHMANIA MEXICANA METACYCLIC PROMASTIGOTES TRANSMITTED TO SKIN BY A SINGLE SAND FLY BITE AND THE HOST TISSUE RESPONSE BY RTQPCR GIRAUD, E.; MARTIN, O.; ROGERS, M. LONDON SCHOOL OF HYGIENE AND TROPICAL MEDICINE, LONDON, UNITED KINGDOM.

Keyword:rtqpcr; metacyclics; infected bite site

Abstract: To quantify the dose of Leishmania mexicana parasites, the proportion of metacyclic promastigotes and the tissue immune response following transmission we developed a strategy using a very sensitive quantitative analysis of both Leishmania and mouse transcripts on a bite site.

A real-time reverse transcription quantitative PCR (RTqPCR)-based method was optimised to analyse parasite and mouse transcripts on the same bite site. One hour post- blood-feed, bite sites were collected and processed for RNA extraction. Using the Leishmania gene target ssrRNA and the metacyclic-specific transcript sherp (small hydrophilic ER-associated protein) we could accurately detect 1 transmitted promastigote and confidently differentiate a range of metacyclic doses (from 75% to 100%) for 5 or more transmitted parasites. Our results with individual sand fly bites show that the majority of flies transmit relatively homogenous populations of metacyclic promastigotes (78% delivered 95-100% metacyclics per bite). Closer analysis revealed high dose transmissions (> 1,000 promastigotes) contained more nonmetacyclic promastigotes (56% delivered < 95% metacyclics per bite) compared to low-dose transmissions ( 1,000 promastigotes; 13% delivered <95% metacyclics per bite). In addition, using the chemokines ccl3 and cxcl2 we found that infected bites attracted 4.4 and 3.8 more macrophages and neutrophils to the bite site, respectively, compared to uninfected fly bites which increased significantly with numbers of parasites inoculated (p < 0.05).

These results describe a versatile method capable of analysing the composition of the transmitted dose and the corresponding host tissue or immune response to infection.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [760] P 792 - COMPARATIVE STUDY OF THE ABUNDANCE OF LUTZOMYIA LONGIPALPIS IN 3 CITIES WITH CANINE VISCERAL LEISHMANIASIS TRANSMISSION IN ARGENTINA. SALOMON, O.D.1; GOULD, I.T.2; SANTINI, M.S.2 1.INSTITUTO NACIONAL DE MEDICINA TROPICAL, PUERTO IGUAZÚ, ARGENTINA; 2.CENTRO NACIONAL DE DIAGNÓSTICO E INVESTIGACION DE ENDEMOEPIDEMIAS, CIUDAD AUTÓNOMA DE BUENOS AIRES, ARGENTINA.

Keyword:canine visceral leishmaniasis; lutzomyia longipalpis; abundance

Abstract: The cities of Clorinda (Formosa), Iguazú (Misiones) and Chajarí (Entre Ríos) are locatedin the area with vectorial transmissiom canine visceral leishmaniasis (CVL) in Argentina. The three cities belong to three eco-regions: Clorinda in the "Chaco" region on the border with Paraguay.

The city of Iguazú, in "Paranaense" region on the border with Paraguay and Brazil, and Chajarí in the "Espinal" region on the border with the Republic of Uruguay. The city of Clorinda has the first record Lu. longipalpis for Argentina associated with urban CVL, in 2004, whereas in Iguazú and Chajarí Lu. longipalpis was reported for the first time in 2010, Chajari in turn being the southernmost record of the distribution of vector for Argentina up to now. This study compared the abundances of Lu. longipalpis between these three cities with transmission of CVL, at the same time analyzing whether differences exist in relation to Lu. longipalpis morphotypes between the three eco-regions. The captures were performed during March 2012 with mini CDC light traps placed in the peridomestic environments for two consecutive nights, following the “worst scenario” criterion. A total of 3829 sand flies of Lu. longipalpis were collected. Iguazú were sampled at 51 sites, while Clorinda and Chajarí were sampled at 20 and 10 sites respectively. The percentage of positive sites obtained was highest in Iguazú 92%, followed by Clorinda 80% and Chajarí 50%, Total abundance of was higher in Iguazú, 2897 individuals, followed by Clorinda with 700 and Chajarí with 309 . This can also be observed for the most abundant sites, Iguazú obtained the highest with 631 individuals, Clorinda and Chajari follow with 543 and 221 respectively. These results showed a different availability of sites with conditions (climate, macro and microhabitat characteristics) for phlebotominae reproductive success, although the different effort of capture and different size of cities could introduce a bias in the same. Lu longipalpis morphotype was also analyzed for the first time for Argentina, The results show that 100% of the specimens observed have two light spots located in the tergites 3 and 4 (morphotype 2 S), despite the eco-region (Chaco, Paranaense and Espinal).

- RESERVOIRS [769] P 793 - A COMPARATIVE ANALYSIS OF DPP TEST WITH ELISA AND IFAT FOR THE DIAGNOSIS OF CANINE VISCERAL LEISHMANIASIS IN BRAZIL. LAURENTI, M.D.1; TOMOKANE, T.Y.1; CARVALHO, A.K.1; SILVA, R.M.2; LAUTENSCHLA, H.R.2; MATTA, V.L.1; SOUZA, C.3; CORBETT, C.E.P.1; MARCONDES, M.4 1.MEDICAL SCHOOL-USP, SAO PAULO, SP, BRAZIL; 2.INSTITUTO ADOLFO LUTZ, SAO PAULO, SP, BRAZIL; 3.FUNDAÇÃO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL; 4.UNESP, SAO PAULO, SP, BRAZIL.

Keyword:canine leishmaniasis; diagnosis; serology

Abstract: The diagnostic screening test for canine visceral leishmaniasis (canVL) recommended by the Brazilian Ministry of Health were ELISA-BioManguinhos (ELISA-Bio) and IFATBioManguinhos (IFAT-Bio), until 2011. However, after the development of a new rapid immunochromatographic test (DPP) by BioManguinhos, it has been used instead IFAT. The aim of this study was to investigate the performance of the DPP test, and compare it with ELISABio, IFAT-Bio, L.chagasi-ELISA and L.chagasi-IFAT, two in house test standardized in our

laboratory using Leishmania (L.) infantum chagasi promastigotes as antigen. For this purpose, sera of 66 dogs with infection confirmed by immunohistochemistry (IHC), and 18 negative controls (healthy animals living outside endemic area) were tested. To detect cross-reactivity, sera of dogs with ehrlichiosis (n=17), babesiosis (n=9), toxoplasmosis (n=9), neosporosis (n=6), neosporosis/toxoplasmosis co-infection (n=4), Chagas disease (n=6) and dirofilariosis (n=4), with no Leishmania infection, were also tested. The sensitivity of the DPP test (93.9%) was equal to ELISA-Bio (93.9%), slightly inferior to L.chagasi-ELISA (96.9%), IFAT-Bio (97.9%) and L.chagasi-IFAT (97.7%). The specificity was higher in both DPP test (94.4%) and L.chagasi-ELISA (97.2%) compared to ELISA-Bio (75%), IFAT-Bio (66.7%) and L.chagasiIFAT (50.8%). Strong cross-reactivity was observed on IFAT-Bio and L.chagasi-IFAT with sera from dogs with Chagas disease (67 and 100%), toxoplasmosis (33 and 44%), neosporosis (50 and 100%) and ehrlichiosis (71 and 29%), respectively. L.chagasi-IFAT presented 100% of cross-reactivity with sera from dogs with neosporosis/toxoplasmosis co-infection. ELISA-Bio showed cross-reactivity especially with sera from dogs with Chagas disease (50%), babesiosis (33%) and ehrlichiosis (35%). DPP and L.chagasi-ELISA presented, respectively, 44 and 22% of cross-reactivity with sera from dogs with babesiosis. Agreement was excellent (k=0.93) between DPP and L.chagasi-ELISA, moderate (k=0.67) when both tests were compared to ELISA-Bio and discreet compared to IFAT-Bio (k=0.47) and L.chagasi-IFAT (k=0.37). Besides the advantage of being a quick test, the DPP showed better performance than ELISA-Bio, IFATBio and L.chagasi-IFAT especially concerning cross-reactivity, and provided very similar results to the L.chagasi-ELISA that used the specific causing pathogen of canVL, as antigen.

Supported by LIM-50 HCFMUSP, FAPESP and CNPq. - VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [771] P 794 - STUDIES ON THE SANDFLY FAUNA (DIPTERA: PSYCHODIDAE) IN AN EMERGING FOCUS OF CUTANEOUS LEISHMANIASIS IN THE BROKOPONDO DISTRICT OF SURINAME KENT, A.D.1; VASCONCELOS, T.2; ADAMS, E.3; MANS, D.4; SCHALLIG, H.3; SAMJHAWAN, A.5 1.DEPARTMENT OF PARASITOLOGY, ANTON DE KOM UNIVERSITY OF SURINAME, PARAMARIBO, SURINAME; 2.EVANDRO CHAGAS INSTITUTE, BELEM, PA, BRAZIL; 3.ROYAL TROPICAL INSTITUTE, AMSTERDAM, NETHERLANDS; 4.DEPARTMENT OF PHARMACOLOGY, ANTON DE KOM UNIVERSITY OF SURINAME, PARAMARIBO, SURINAME; 5.PUBLIC HEALTH SERVICE, DEPARTMENT OF ENTOMOLOGY, PARAMARIBO, SURINAME.

Keyword:cutaneous leishmaniasis; vectors of leishmania, leishmaniasis in suriname; psychodidae, sandfly fauna

Abstract: Background In Suriname, cases of cutaneous leishmaniasis (CL) have increased over de past decade and are becoming a public health problem, according to the dermatology Service. Also Leishmania species other than L (V.) guyanensis have been found such as L (V.) braziliensis, L (L.) amazonensis, and L (V.) naiffi. Transmission occurs mainly in the forested hinterland through the bite of an infected female sandfly of the genus Lutzomyia. Due to an increase in activity in the hinterland because of gold mining, tourism and logging, many people travel to these areas with a high risk of getting infected. The district of Brokopondo is one of the high risk transmission areas in the hinterland because of high mining activities, the presence of tourist camps and the major of road leading to the interior passes through this district.

Aim

The last sand fly survey in Suriname is dates from the period between 1952 and 1984, and therefore information regarding the potential vectors of leishmaniasis in Suriname is limited.

This study aims to broaden the knowledge of vectors of medical importance involved in the transmission of CL in one of the foci in Suriname.

Methods/principal findings Sandflies were collected using CDC light traps in the period between September 2010 and July 2011, mainly in the rainy season. The survey was performed in areas around mining plots and villages. A total of 665 sandfly specimens were captured belonging to 21 different species. The most abundant species were L. ubiquitalis (37.8%), L. squamiventris maripaensis (4.5%), L. ininii (3.6%), L. longipennis (2.7%), and L. infraspinosa (1.8%). Also other known vectors of Leishmania were captured: L. umbratilis (0.8%), L. flaviscutellata (0.6%) and L. whitmani (0.3%). In addition 2 not previously recorded species for Suriname were captured: L. ayrozai and L. carvalhoi.

Conclusion This study provides an update of the sandfly diversity of the district Brokopondo and recorded 2 new species for Suriname of which 1(L. ayrozai) is a known vector of Leishmania in other Latin-American countries.

- EPIDEMIOLOGY [774] P 795 - GEOGRAPHICAL DISTRIBUITION OF CANINE VISCERAL LEISHMANIASIS IN INDIGENOUS LAND XAKRIABÁ, MINAS GERAIS, BRAZIL. MACHADO-COELHO, G.L.L.1; ROCHA, A.M.S.1; FRANCA-SILVA, J.C.2; SILVA, J.C.3; REIS, V.W.1; SANTOS, R.C.1; BATISTA, A.P.1; FONSECA, A.M.2; CELESTE, J.L.2; MARINHO, C.C.1; BAHIA, M.T.4; FERREIRA, E.C.5; ANDRADE, H.M.2 1.ESCOLA DE MEDICINA, UNIVERSIDADE FEDERAL DE OURO PRETO, OURO PRETO, MG, BRAZIL; 2.INSTITUTO DE CIÊNCIAS BIOLÓGICAS, UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 3.SECRETARIA ESPECIAL DE SAÚDE INDÍGENA, MINISTÉRIO DA SAÚDE, BELO HORIZONTE, MG, BRAZIL; 4.INSTITUTO DE CIÊNCIAS EXATAS E BIOLÓGICAS, UNIVERSIDADE FEDERAL DE OURO PRETO, OURO PRETO, MG, BRAZIL; 5.FUNDAÇÃO OSWALDO CRUZ, UNIDADE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL.

Keyword:canine visceral leishmaniasis; prevalence; xakriabá

Abstract: Visceral leishmaniasis (VL) in Brazil, is an endemic peri-urban and rural zooantroponose. This disease is potentially fatal in humans whith a mortality up to 10% when no appropriate treatment is instituted. According to the Brazilian Ministry of Health, the expansion of this zoonosis has affected individuals with co-morbidities such as malnutrition and low immunity, which has caused a high number of deaths. The northern region of Minas Gerais state there is a high notification rate of human cases. Dog is an important domestic reservoir, and is the major responsible for the VL endemic. In the indigenous land Xakriabá (ILX), the prevalence of mild to severe malnutrition among children < 13 years old is high. Thus the aim of this study was to evaluate the seroprevalence of canine visceral leishmaniasis (CVL) in ILX. A census serological survey was conducted in 17 of the 32 villages of ILX. Blood dried on filter paper was collected from 949 dogs and sent weekly to the Laboratory of Epidemiology of Parasitic Diseases, School of Medicine of Federal University of Ouro Preto in plastic bags containing silica gel. The eluate obtained was used to perform the ELISA and indirect immunofluorescence assay (IFA). 33.7% of the samples were positive for ELISA and 7.6% positivity in IFA. The prevalence of CVL was heterogeneous for villages, ranging from 14.6 to 60.0% for ELISA and from 1.3 to 32.8% for

IFA. Dogs seropositive for both tests were euthanized (n=35) and 24 necropsied. Of these, 23 (95.8%) had clinical symptoms of LVC: onychogryphosis or splenomegaly (91.3%), lymphadenopathy (90.5%) and weight loss (30.4%). We collected spleen, lymph nodes, skin and bone from the necropsied dogs, and the samples were sent frozen in liquid nitrogen to the Laboratory of Leishmaniasis and Vaccines, Biological Science Institut of Federal University of Minas Gerais. The marrow culture was positive for promastigotes of Leishmania sp. in 12 dogs (57.1%) and the species isolated was Leishmania (L.) infantum chagasi. The prevalence of LVC in this indigenous area is high, and considering the vulnerability of the child population, ILX should be classified as an area of permanent surveillance. Financial support: CNPq (481001/2010-7), FAPEMIG (PPM-00154-12) e CAPES (23038.007082/2011-52).

- EPIDEMIOLOGY [775] P 796 - RELATIONSHIP BETWEEN THE OCCURRENCE OF VISCERAL LEISHMANIASIS AND CLIMATE VARIABILITY IN A HIGHLY ENDEMIC AREA OF BANGLADESH RAHMAN, M.W.; HUDA, M.; MOTALAB, M.A.; RAHMAN, M.; LUBY, S.P.; STREATFIELD, P.K.; RAHMAN, K.M. INTERNATIONAL CENTRE FOR DIARRHOEAL DISEASE RESEARCH, BANGLADESH, DHAKA, BANGLADESH.

Keyword:visceral leishmaniasis; occurance; climate variability

Abstract: Introduction: Visceral Leishmaniasis (VL) is endemic in 43 out of 64 districts in Bangladesh.

Robust community surveillance has estimated VL incidence in highly endemic areas in Bangladesh. It is also among the most vulnerable countries affected by climate change. How climate variability affects the occurrence of VL in Bangladesh is yet to be evaluated. We aimed to determine relationships between changes in climate parameters and the occurrence of VL in a highly VL endemic area in Bangladesh.

Methods: We conducted an ecological study. We obtained data on climate parameters including temperature, rainfall, humidity and cloud cover from 2000 to 2009 in Mymensingh District from the Bangladesh Meteorological Department. Details of VL cases reported at Fulbaria Upazila Health Complex (UHC) in Mymensingh District for the same period were extracted from UHC records. We used scatter plots to observe seasonality and monthly peaks of VL cases against different climate parameters. Due to over dispersion, a generalized linear negative binomial regression model was developed using monthly VL cases and climate parameters. We included indicator variables for each month and year respectively to account for seasonality, long term trends, or other variations. To adjust autocorrelations, we added an autoregressive term (at order one) to the model. We adjusted for potential mutual confounding between the four parameters to identify independent associations of monthly VL cases and individual climate parameter.

Results: There were several seasonal peaks of monthly VL cases, the highest in June-July every year. Major annual peaks were found in 2005, 2006 and 2008. VL cases in Fulbaria increased with increased humidity (p<0.001) and rainfall (p=0.040) with lag 0-6 months. VL occurrence did not vary considerably with changes in temperature (p=0.068) and cloud-cover (p=0.394).

However after adjusting for mutual confounders among monthly VL cases and other climatic parameters, only humidity showed significant association (p=0.039). Each 1% increase in humidity was associated with a 4% increase in the number of VL cases after 6 months.

Discussion: Humidity was the only contributing factor for increasing monthly VL occurrence in our study. The wide range of lag period may be due to the variation of the incubation period of VL as well as differential care seeking delay of patients. Further studies on vector density are needed to reconfirm the findings of our study.

- EPIDEMIOLOGY [777] P 797 - RISK LEVELS OF LOCALIZED CUTANEOUS LEISHMANIASIS PER GEOPOLITICAL ENTITY, VENEZUELA, 2006-2010 DE LIMA, H.; BORGES, R.; SALGADO, A.; CONVIT, J. INSTITUTO DE BIOMEDICINA, CARACAS, VENEZUELA; .

Keyword:leishmaniasis cutaneous; epidemiology; venezuela

Abstract: In Venezuela leishmaniasis is a public health problem. In the 2006-2010 period 11,376 cases of localized cutaneous leishmaniasis (LCL) were registered, reflecting an average annual rate of 8.15 cases per 100,000 inhabitants (2,275 cases per year). The objective of this study was to determine risk areas for LCL according to the geopolitical divisions of the country (states, municipalities and parishes), and quantify the exposed population. This was done by calculating the average annual rate (AAR) of the quinquennium under study per 100,000 inhabitants. A scale was developed with five risk levels defined by the following parameters: No Risk (NR), AAR of 0.5 or less; Very Low Risk (VLR), AAR of 0.51 to 5; Low Risk (LR), AAR between 5.01 and 10, Moderate Risk (MR), AAR between 10.01 and 100, High Risk (HR), AAR over 100. Populations were estimated according to the National Statistics Institute (INE), 2001 Census. Based on the criteria used none of the states were considered HR, 9(37.5%) states were MR, while at the other end only 4 of them can be considered NR. At parish level, the smallest political division, was observed that of the existing 1,116, 32 are HR with an AAR ranging between 100 and 450 cases per 100,000 inhabitants; these represent 2.87% of the parishes with a population of 353,655 inhabitants (1.27% of the total population). The average population per parish in HR areas is 11,051.70, so apparently these are relatively small parishes with regard to the number of inhabitants. HR Parishes are distributed in 13 states, primarily associated with mountainous or Andean foothills zones. On the other end of the scale, 578 (51.79%) parishes were found to have no LCL risk according to the criteria used; 14,365,690 persons inhabit these, ie 51.54% of the general population. In these parishes the average population per parish is 24,854.14 inhabitants. 539 (93.25%) NR parishes did not report a single case during the period.

Conclusions: This analysis allows the identification of high or moderate endemicity areas where the implementation of preventive and control measures could be more effective, resulting in greater benefits for affected populations. The analysis of the results at state level does not permit the identification of HR areas, to achieve this the analysis must be carried out at the lowest levels of disaggregation (parishes). A tendency is observed that the average of inhabitants in HR parishes is lower than in parishes where there is no risk of disease.

- EPIDEMIOLOGY [785] P 798 - STUDY OF VECTORS AND HOST OF LEISHMANIA IN VISCERAL AND CUTANEOUS LEISHMANIAISIS AREAS IN CENTRAL WESTERN OF MINAS GERAIS STATE – BRAZIL SILVA, E.S.1; TEIXEIRA NETO, R.G.2; NASCIMENTO, B.W.L.3; TONELLI, G.3; BOTELHO, H.A.2; ANDRADE FILHO, J.D.3; GONTIJO, C.M.F.2

1.UNIVERSIDADE FEDERAL DE SÃO JOÃO DEL REI – CAMPUS DONA LINDU, DIVINÓPOLIS, MG, BRAZIL; 2.LABORATÓRIO DE LEISHMANIOSES – CENTRO DE PESQUISAS RENÉ RACHOU – FIOCRUZ/MG, BELO HORIZONTE, MG, BRAZIL; 3.CENTRO DE REFERÊNCIA NACIONAL E INTERNACIONAL PARA FLEBOTOMÍNEOS – FIOCRUZ/MG, BELO HORIZONTE, MG, BRAZIL.

Keyword:reservoirs; vectors; epidemiology

Abstract:

Leishmaniasis is a parasitic disease caused by protozoa of the Leishmania genus, which are transmitted by phlebotomine sand flies to different vertebrate hosts. Over the last decade, the geographical spread and the number of cases of visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) has increased considerably within Brazil, making it one of the most serious problems faced by the public health authorities.

Divinópolis municipality is considered an endemic region of CL. From 2000 to 2011, 52 cases were recorded by Health Authorities. Considering the occurrence of VL in the municipality, 15 cases were registered in the last 5 years, being considered a moderated transmission area.

Disorganized growth of Divinópolis can favor the proximity of suburban and residual forest areas. This fact can promote the occurrence of transmission cycles of leishmaniasis.

Nevertheless there are no concluded studies about the epidemiology of leishmaniasis in the municipality.

The aim of this study was provide information about sand flies vector and wild and synanthropic host animals in urban and green areas of Divinópolis municipality.

Sampling of vectors was performed every month between September 2010 an August 2011, using CDC light traps (HP model), located in fifteen houses and five green areas in the municipality. To capture small rodents and marsupials traps were distributed across transects in the green areas (twenty traps were placed along each trail) and in the same fifteen houses (two traps per house). Catches of small mammals were held every two months.

Presence of the parasite was also investigated by amplifying DNA isolated directly from the tissue samples of small mammals and sand flies. Different targets of Leishmania DNA, hsp70 and internal transcribed spacer ITS1, were used to detect and identify the parasite.

A total of 102 specimens of small mammalian and 1064 sand flies were collected. Urban sites displayed a much higher abundance of phlebotomines than the green areas (83,7% and 16,3% respectively). However abundance of small mammalian was higher in green areas (97,1%).

A total of three specimens of small mammalian was positive in the PCR for leishmania, one Didelphis albiventris and one Necromys lasiurus infected by L. braziliensis, and one Rattus rattus infected by L. infantum. None sandfly was positive in PCR reactions analyzed, but from the 17 collected species, seven are proven or suspicious vectors of Leishmania. The main vector of L. infantun, L. longipalpis was the most abundant species (76,9%).

Further studies will be developed and spatial analysis of vector and host may provide important information for understanding the epidemiology of leishmaniasis in this study area.

- CONTROL PROGRAMS [826] P 799 - DECREASE OF CANINE VISCERAL LEISHMANIASIS WITH THE USE OF DOG COLLARS IMPREGNATED WITH DELTAMETHRIN ZUQUE, F.R.S.1; ZUQUE, M.A.S.2; ZUQUE, F.T.S.2; SILVA, I.S.3 1.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL - UFMS / CPCX, COXIM, MS, BRAZIL; 2.FACULDADES INTEGRADAS DE TRÊS LAGOAS - AEMS, TRÊS LAGOAS, MS, BRAZIL; 3.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL.

Keyword:canine visceral leishmaniasis; dog collar impregnated with deltamethrin; control measures

Abstract:

Introduction: The introduction of the etiological agent from domestic reservoirs in areas infested

by Lutzomyia longipalpis, seems to be a facilitator of urbanization Visceral Leishmaniasis (VL).

For vector control is used with household insecticide spraying of pyrethroids class; however, other alternative and complementary measures have also been used as topical treatment of dogs with insecticides and use of dog collars impregnated with deltamethrin 4% . Objective: To evaluate the prevalence of Canine Visceral Leishmaniasis (CVL) after the use the dog collars impregnated with deltamethrin 4% in the city of Tres Lagoas - MS (Brazil) in 2008 and 2009.

Methodology: The study was based on secondary data analysis of Surveillance, Center for Zoonosis Control of Endemic Diseases sector and the city of Tres Lagoas. We evaluated the results of interventions in 10 sectors in the area of intense transmission, in 2008 and 2009.

Results: In 2008 the sectors were sprayed, 1, 2, 5 and 10 with presence of 988 dogs in residency, of which 83% (n = 816) received the collar and 172 were collected for euthanasia to be reactive serum. In 2009 nine sectors were sprayed with the presence of dogs in 3005 in residency and 19% (n = 561) were reactive serum being collected for euthanasia and the others received collars. Only sector 7 was without spraying these two years using only the collars as an alternative measure. Of the four sectors that received the spray and used the collar in two consecutive years, there was a reduction in the prevalence of CVL in sectors 2 and 5: the 156.250 / 00 142.860 / 227.090 00 and / 00 to 160.870 / 0 respectively. However, in sectors 1 and 10 there was an increase in the prevalence of CVL. By comparing the prevalence of LVC from 2008 to 2009 showed that the sectors 3, 4, 6, 8 and 9 before distribution of collars prevalence ranges from 178,95 0/00 to 406,78 0/00 and further to the distribution of Collars ranged from 114,290/00 to 258,160/00 . In Sector 7 where only the collars were used, the prevalence of CVL ranged from 202900/00 in 2008 to 190.750/00 in 2009. The prevalence of CVL decreased in 8 of the 10 sectors in the area of intense transmission. Conclusion: Despite the cost of the dog collar impregnated with deltamethrin to 4%, their continued use in dogs no reagents, coupled with measures recommended control was efficient in reducing the prevalence of CVL in the city.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [827] P 800 - A REVIEW OF NATURAL LEISHMANIA INFECTION OF SAND FLIES IN BRAZIL REGO, F.D.; FILHO, J.D.A.; GONTIJO, C.M.F.; SHIMABUKURO, P.H.F.; CARVALHO, G.M.L. CENTRO DE PESQUISAS RENĂ&#x2030; RACHOU, BELO HORIZONTE, MG, BRAZIL.

Keyword:sand flies; infection; leishmaniases

Abstract: Phlebotomine sand flies transmit pathogens to humans and non-human animals worldwide, including the protozoan Leishmania. Different clinical manifestations of leishmaniases arise from infections with different species of Leishmania, which in turn can be transmitted by different species of sand flies. There are over 800 species of sand flies recorded in the world, of which 98 are proven or suspected vectors of human leishmaniases; these include 56 species in the New World involved in the transmission of 15 Leishmania species. Current studies provide approximate numbers of sand flies species incriminated or suspected in the transmission of leishmaniases, however, systematic data concerning the actual number of species involved, the technique used for the detection, the location of the study and the correlation parasite / vector are not yet available. Thus, the availability of such a database is extremely important to help elaborate future studies, since the disease occur in all states of Brazil. Here, we review the number of sand flies species incriminated in the transmission of Leishmania since the decade of 1970 and the findings of natural infection in sand flies captured in Brazil. We have found a total of 44 species belonging to 10 genera involved in the transmission of leishmaniases: Bichromomyia (3), Evandromyia (5), Lutzomyia (4), Micropygomyia (2), Migonemyia (1), Nyssomyia (8), Pintomyia (5), Psathyromyia (3), Psychodopygus (12) and Trichophoromyia (1).

Our results showed that two genera, Psychodopygus and Nyssomyia presented more species found with Leishmania, 12 and 8 species, respectively. The latter genus presents infection with six confirmed species of Leishmania, while Psychodopygus had only two confirmed species of this parasite. Dissection of the digestive tract of the insect with subsequent characterization of the parasite was the most used method in papers published until the 80’s. Given the difficulty of dissection techniques and the continuous improvement of molecular tools, there has been an increase in the number of works based on the detection of parasite DNA. According to the data we compiled, only a small number of species fit the criteria for vector incrimination and few studies were conducted to establish the actual role of these species in the epidemiological cycle of leishmaniasis.

- RESERVOIRS [832] P 801 - FIRST RECORD OF LEISHMANIA (VIANNIA) BRAZILIENSIS IN BATS (CHIROPTERA) IN MATO GROSSO DO SUL, SOUTHWEST BRAZIL: NEW CHALLENGES FOR THE CONTROL OF LEISHMANIASIS IN AN ENDEMIC AREA? SHAPIRO, J.T.; LIMA JUNIOR, M.S.C.; DORVAL, M.E.C.; FRANÇA, A.O.; BORDIGNON, M.O. UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL.

Keyword:reservoirs; leishmaniasis; zoonosis

Abstract: Leishmaniasis is a zoonotic disease caused by parasites of the genus Leishmania that has expanded beyond its natural range and is becoming increasingly urban, changes linked to environmental degradation. Although Leishmania (Viannia) braziliensis has been detected in dogs, cats and horses, in some areas, there is no apparent correlation between infections in humans and domestic animals, pointing to the role of wild fauna reservoirs in the maintenance and transmission of leishmaniasis. Currently the most likely candidates for wild reservoir species are small mammals such as rodents and marsupials. The objective of this study was to detect Leishmania sp. in the bats of Campo Grande, Mato Grosso do Sul, an endemic area.

Using PCR and PCR-RFLP, we identified Leishmania (Viannia) braziliensis in a blood sample, a lesion and a slide imprint of liver from two bats (Chiroptera). The animals testing positive were found in both a rural site and an urban site, indicating that bats in various environments in the region may be carrying the parasite and could play a role as sylvatic reservoirs. This is the first register of Leishmania in bats in the state and the first register of L. (V.) braziliensis in bats.

Given the large populations of urban bats, their mobility, and their ability to come in close contact with humans, these results could have serious implications for public health and the control of leishmaniasis in Mato Grosso do Sul. The small number of studies on Leishmania in bats indicates a need for further investigation into the role they may play as potential reservoirs of the parasite and possibly establishing urban cycles of transmission of the disease.

- EPIDEMIOLOGY [833] P 802 - MOLECULAR IDENTITY AND THE DRUG SENSITIVITY PROFILES OF RECENT CLINICAL ISOLATES OF KALA-AZAR FROM INDIA AND BANGLADESH MANNA, M.1; KHANRA, S.2; MONDAL, D.3; SAHA, P.4; BANDOPADHYAY, S.K.5; ROY, S.6 1.BARASAT GOVT. COLLEGE, BARASAT, INDIA; 2.BETHUNE COLLEGE, KOLKATA, INDIA; 3.INTERNATIONAL CENTRE FOR DIARRHEAL DISEASE RESEARCH, DHAKA, BANGLADESH; 4.SAHA INSTITUTE OF NUCLEAR PHYSICS, KOLKATA, INDIA; 5.CALCUTTA NATIONAL MEDICAL COLLEGE, KOLKATA, INDIA; 6.INDIAN INSTITUTE OF CHEMICAL BIOLOGY, KOLKATA, INDIA.

Keyword:indian kala-azar, ; restriction fragment length polymorphism; its1 sequencing

Abstract:

Visceral Leishmaniasis or Kala-azar (KA) is a serious health problem in India. Characterization of Leishmania sp. is tremendously important to understand the epidemiology, taxonomy and population genetics of the parasites which eventually helps in designing suitable drug regimen to conflict the disease. In this study, we typed the clinical isolates [n=15] of Leishmania species collected in the period 2006–2010 from patients diagnosed with Kala-azar and Post Kala-azar dermal leishmaniasis (PKDL) by RAPD-PCR method using eight selected primers and Restriction Fragment Length Polymorphism (RFLP) of three genetic markers viz., Internal Transcribed Spacer (ITS), ITS1 and heat shock protein 70 (hsp70). Experimentally, ITS, ITS1 and hsp70 regions of genomes of all the clinical isolates were separately amplified by PCR and then digested with restriction enzymes: ITS with Alu1, EcoR1 and Msp1, ITS1 with Hae III and Rsa1 and hsp70 with Hae III. The RAPD profiles and the RFLP profiles of the clinical isolates were compared with that of the WHO reference strains for Leishmania donovani (DD8) and Leishmania tropica (K27), respectively. The ITS1 regions of all the clinical isolates along with the two WHO reference strains were sequenced and a phylogram was constructed. Interestingly, the RAPD and the RFLP profiles and phylogram revealed closeness of one isolate (T5) with K27. This observation is in congruence with an earlier report claiming that both the species are responsible for KA in India and thus, emphasizes urgent need for systematic characterization of the clinical isolates of Indian KA as appropriate treatment administration relies primarily on proper diagnosis. We have also checked the drug sensitivity of the clinical isolates for the drugs Sodium Stibogluconate (SSG) and Miltefosine respectively. The clinical isolate, T5 is unresponsive to SSG (Sodium Stibo Gluconate) but sensitive to Miltefosine.

- RESERVOIRS [839] P 803 - SEROLOGICAL REACTIVITY TO CANINE LEISHMANIASIS IN TRIÂNGULO MINEIRO AND ALTO PARANAÍBA REGION, MINAS GERAIS, BRAZIL BATISTA, L.R.; TIBÚRCIO, M.G.S.; HERNÁNDEZ, C.G.; SILVA, M.V.; COSTA, T.Á.; ASSIS, E.W.S.D.V.; JÚNIOR, V.R.; RAMIREZ, L.E. UNIVERSIDADE FEDERAL DO TRIÂNGULO MINEIRO, UBERABA, MG, BRAZIL.

Keyword:canine leishmaniasis; elisa; leishmania

Abstract: Leishmaniasis is caused by protozoan parasites of Leishmania genus, and is characterized as a complex of diseases with a large spectrum of clinical and epidemiological diversity. Visceral leishmaniasis has the dog as the main reservoir of the parasite, especially in South America, where a high level of canine infection is associated with an increased risk of human disease.

Ministry of Health recommends serology (ELISA, DPP) as a diagnostic method of canine visceral leishmaniasis. However, ELISA can result in false positives, especially for crossreactivity with other parasites, such as Trypanosoma cruzi, dermotropic Leishmania species, Babesia canis and Erlichia canis. In Minas Gerais, the disease is expanding. In Triângulo Mineiro, despite to be endemic for Chagas disease, is still considered non-endemic for visceral leishmaniasis. 229 Serum samples were collected from dogs from Uberaba (128), Delta (15), Água Comprida (58), Nova Ponte (13) and Perdizes (15), cities belonging to Triangulo Mineiro and Alto Paranaiba region. Other biological samples from these animals had been previously tested by PCR for Leishmania spp. and T. cruzi. Antibodies anti-Leishmania were determined in a home made ELISA, using soluble crude antigen from promastigote forms of L. infantum. Cutoff was calculated as the mean optical density (OD) for negative control sera plus five standard deviations. Antibody titers were expressed as ELISA index (EI) as follows: EI=ODsample/ODcut. EI > 1.2 were considered positive. In Uberaba, 113 (88.3%) of the 128 dogs were from the Zoonosis Control Center (CCZ). 41 dogs (32%) were considered positive for ELISA, 37 of them from the CCZ. Reactive test were: 9 dogs (60%) in Delta, 15 dogs

(25.9%) in Água Comprida, and only one dog (7.7%) in Nova Ponte. In Perdizes, any dog was reactive to the test. Of the 66 positive dogs (28.8%), 7 (10.6%) were PCR positive for T. cruzi and 2 (3%) were PCR positive for Leishmania spp. In addition, 16 dogs were PCR positive for T. cruzi, but there were not reactive in ELISA. Although still not considered an endemic area, Triângulo Mineiro and Alto Paranaiba region is located next to endemic areas, in addition to being the target of people from endemic regions migration. These results demonstrate that studied region could present non autochthonous Leishmania spp. infected dogs, facilitating the spread and establishment of the transmission cycle in these municipalities, especially in the presence of the sand fly.

- EPIDEMIOLOGY [842] P 804 - EPIDEMIOLOGICAL PROFILE OF PATIENTS DIAGNOSED WITH AMERICAN CUTANEOUS LEISHMANIASIS IN THE CENTER OF TROPICAL MEDICINE OF RONDONIA, PORTO VELHO, BRAZIL FERNANDES, L.S.1; JUNIOR, C.F.S.2; BAIÃO, A.1; AZEVEDO, F.S.1; CANTANHÊDE, L.M.3 1.FEDERAL UNIVERSITY OF RONDONIA, PORTO VELHO, RO, BRAZIL; 2.FEDERAL UNIVERSITY OF RONDONIA AND CENTER OF TROPICAL MEDICINE OF RONDONIA, PORTO VELHO, RO, BRAZIL; 3.OSWALDO CRUZ FOUNDATION, PORTO VELHO, RO, BRAZIL.

Keyword:epidemiology; american cutaneous leishmaniasis; rondonia

Abstract: Brazil is one of the countries with the highest number of cases of American Cutaneous Leishmaniasis (ACL), especially the North, Northeast and Midwest. This made the tropical disease an endemic zoonosis in the Western Amazon and affects many people every year in these locations and worldwide. This study aims to analyze the epidemiological profile of 58 patients diagnosed with American Cutaneous Leishmaniasis and attended in the Center of Tropical Medicine of Rondonia (CEMETRON), in the municipality of Porto Velho. It is an observational descriptive case series that occurred from June 2011 to December 2012. During this period, were collected data about the gender, age, profession, nationality, origin and others socioepidemiological and clinical data related to the disease, through an organized form by the responsible physician. Of the patients studied, the average age was 43 years and the vast majority, 82.75% (47) were males. The most frequently profession found was agricultural worker, with 30% (17), and in other 15.52% (9) of the occupations had contact with forest areas, workers as hydroelectric dams workers, fishermen and miners. As for naturalness, 53.44% (31) of the patients were from the North, and 74.19% (23) of these, were from Rondonia. Others 25% (15) were born in the Northeast (15%) and Midwest (10%). About the origin, 60.34% (35) were from the capital of Porto Velho and surrounding cities and 27.58% (16) in the interior of the state of Rondonia. It was also found that 86.20% (50) of the patients already had some contact with areas of forest at least once in their lives and the same percentage (86.20%) have never traveled abroad. At the end of the study it was concluded that the epidemiological profile more common in patients with ACL treated at CEMETRON is male, around 43 years, and occupations at risk for the disease and allow the contact with forests, rivers, or deforested areas, mostly born and living in areas endemic for the disease (Rondonia, Northeast and Midwest) and who have had contact with forests at least once during their lives.

- CLINICAL LEISHMANIASIS [845] P 805 - CUTANEOUS LEISHMANIASIS IN PORTUGAL: IMPORTED AND ATYPICAL AUTOCHTHONOUS CASES CORTES, S.1; CRISTOVÃO, J.M.2; LOPES, L.3; OLIVEIRA, J.4; MANSINHO, K.5; CAMPINO, L.6 1.INSTITUTO HIGIENE MEDICINA TROPICAL(IHMT)/CMDT-UNIVERSIDADE NOVA DE LISBOA(UNL), LISBOA, PORTUGAL; 2.INSTITUTO HIGIENE E MEDICINA TROPICAL-UNL, LISBOA, PORTUGAL; 3.HOSPITAL DE SANTA MARIA, LISBOA, PORTUGAL; 4.HOSPITAL FERNANDO DA FONSECA, LISBOA, PORTUGAL; 5.HOSPITAL EGAS MONIZ. IINSTITUTO HIGIENE

MEDICINA TROPICAL-UNL, LISBOA, PORTUGAL; 6.IINSTITUTO HIGIENE MEDICINA TROPICAL-UNL. DEP.CIÊNCIAS BIOMED. E MED.-UNIVERSIDADE DO ALGARVE, LISBOA, PORTUGAL.

Keyword:cutaneous leishmaniasis; imported cases; autochthonous cases

Abstract: In Portugal, visceral leishmaniasis (VL) assumes the most frequent form of the disease, however, cutaneous leishmaniasis (CL) has been increasingly detected with about 10 new cases/year. CL is considered benign with self cure but several atypical cases have been observed in recent years, from autochthonous L. infantum infections. Other strains have been identified from imported cases such as L. braziliensis, L. guyanensis and L. major.

In the present work we report atypical autochthonous cases of CL with mucosal lesions and imported CL cases.

Authoctonous cases: 1) A male adult previously diagnosed with VL, presented with nasal obstruction and recurrent epistaxis. On observation a nasal polyp was biopsied and molecular analysis revealed L. infantum DNA; 2) A female adult, presented a facial lesion on the nose and malar region, with local edema, erythema and scaling. Leishmania parasites were identified as L. infantum; 3) A male adult presented a recurrent pustule in the nasal vestibule, and on observation exhibited an erythematous desquamative lesion which subsequently extended to the upper lip. Molecular tests identified L. infantum DNA. Imported cases: 1) A male adult who add been living in Brazil in the past two years, with previous diagnosis of CL, presented various cutaneous ulcers distributed throughout his body.

Parasites were isolated from the lesions and L. guyanensis was identified by isoenzyme typing; 2) A girl who had travelled to Tunisia in the previous months, presented several cutaneous lesions in the face and arms. Paraffinised material was analyzed with molecular markers and L. major was identified; 3) A male adult presented an ulcerative tumoral lesion in the forearm. The patient had traveled to Bolivia the previous year. L. braziliensis was identified.

These cases highlight the need to recognize the disease caused by endemic, even atypical cases, and non endemic Leishmania species. It is important that physicians be cognizant of the possibility of imported tropical diseases, namely cutaneous leishmaniasis, in order to perform early diagnosis, parasite identification and adequate treatment. The risk of introduction and spread of non endemic Leishmania species due to traveling to endemic areas and global migration could not be discarded. In addition these species could be transmitted by sand fly species normally considered non-permissive to Leishmania infection.

S. Cortes holds postdoctoral grant (SFRH/BPD/44450/2008 FCT). campino@ihmt.unl.pt

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [846] P 806 - DISTRIBUTION OF VECTORS OF LEISHMANIASIS IN PUERTO IGUAZÚ, ARGENTINA, IN THE BORDER WITH PARAGUAY AND BRAZIL. SALOMON, O.D.1; SANTINI, M.S.2; FERNANADEZ, M.S.2; GOULD, I.T.2; MANTECA ACOSTA, M.1; BERROZPE, P.E.2; GOMEZ, A.3 1.INSTITUTO NACIONAL DE MEDICINA TROPICAL, PUERTO IGUAZÚ, ARGENTINA; 2.CENTRO NACIONAL DE DIAGNOSTICO E INVESTIGACIÓN DE ENDEMOEPIDEMIAS, CAPITAL FEDERAL, ARGENTINA; 3.FUNDACIÓN MUNDO SANOI, CAPITAL FEDERAL, ARGENTINA.

Keyword:distribution of vectors ; lutzomyia longipalpis ; argentina

Abstract: Lutzomyia longipalpis (visceral leishmaniasis, LV) and Nyssomyia whitmani (cutaneous leishmaniasis, LC) have been reported in the urban area of Puerto Iguazú, Argentina for the first time in 2010, and Lu. longipalpis has been associated with dogs rK 39+. Therefore, the distribution in time of vectors of Leishmania spp. in the city was studied. Puerto Iguazú is located in the northeast of Argentina, in the border area with Paraguay and Brazil. In order to sample the entire urban area, the city was divided into 400m2 patches; one household was selected within each patch using “the worst scenario” criterion. This work was carried out at the beginning of the 4 seasons seasonally (spring, summer, fall, and winter). The light traps were placed in each site for two consecutive nights, except in early spring where they were placed only one night (total sampling effort 355 trap per night placed in 51 peridomiciles). Species abundance of each household was estimated using the number of captured individuals trapnight. Nine species were captured: Lu. longipalpis, Ny. whitmani, Migonemyia migonei, Mi. quinquefer, Evandromyia cortelezzii-sallesi, Pintomyia pessoai, Pi. sordelli, Psathyromyia shannoni, Bruptomyia sp. The first two species summed up 97% of the total sample of 5110 sandflies throughout the year. Ny. whitmani was most abundant in early spring and early summer (11.9, SD 60.1; 7.7, SD 26.4, individuals per trap per night respectively) while Lu. longipalpis was most abundant in early fall (30.1, SD 92.5, individuals trapnight) and it was recorded in 65% of the peridomiciles. Both species were present in all the seasons, even in early winter when the lowest captures were registered. Taking into account the total sample the whole abundance, Lu. longipalpis was more abundant than Ny. whitmani (3796 vs. 1545). While further captures are still ongoing, these preliminary results show a temporal trend of the abundance of vectors in the studied area. Puerto Iguazú is then an area of risk of transmission of Leishmania spp. mainly during the early fall, but additional studies will be necessary to establish the risk of transmission of LV and LC throughout the year. Given its geographical location and the transit between border cities, coordinated strategies for prevention and control of vectors at regional level are required. However, to reduce potential breeding sites, work with the community should be adapted to local level due to cultural differences.

- RESERVOIRS [855] P 807 - SEROLOGICAL REACTIVITY TO CANINE LEISHMANIASIS IN NON-ENDEMIC REGION, ITUIUTABA CITY, MINAS GERAIS, BRAZIL TIBÚRCIO, M.G.S.1; BATISTA, L.R.1; HERNÁNDEZ, C.G.1; SILVA, M.V.1; COSTA, T.Á.1; ASSIS, E.W.S.D.V.1; FARIA, K.F.2; CASTRO, R.O.1; MORAES, P.N.1; JÚNIOR, V.R.1; RAMIREZ, L.E.1; OLIVEIRA, K.R.2 1.UNIVERSIDADE FEDERAL DO TRIÂNGULO MINEIRO, UBERABA, MG, BRAZIL; 2.UNIVERSIDADE FEDERAL DE UBERLÂNDIA, ITUIUTABA, MG, BRAZIL.

Keyword:canine leishmaniasis; elisa; leishmania

Abstract: Leishmaniasis are parasitic diseases with enormous impact in human and veterinary medicine.

The visceral form, considered the most severe, is increasing in Brazil, despite major efforts by scientific authorities. Domestic dog is the main domestic reservoir of Leishmania infantum and has important role in disease transmission cycle to humans. In Minas Gerais state, Northwest and Central regions are the most affected, and has a growing number of cases. However, Triângulo Mineiro region is considered a non-endemic area for visceral leishmaniasis, with a small number of confirmed autochthonous cases. According to Ituiutaba Municipal Helth Department, in 2012 were confirmed 4 canine leishmaniais cases in the city. Ituiutaba city is localized in Pontal do Triângulo Mineiro region, borderland to Goiás state, with predominantly rural activities. 551 dog blood samples were collected onto filter paper (eluate), 173 (33,8%) from rural area and 338 (66,2%) from urban area, during anti-rabic vaccination campaign in

2012. Antibodies anti-Leishmania were determined in a home made ELISA, using soluble crude antigen from promastigote forms of L. infantum. Cut-off was calculated as the mean optical density (OD) for negative control sera plus five standard deviations. Antibody titers were expressed as ELISA index (EI) as follows: EI = ODsample/ODcut. EI > 1.2 were considered positive. 169 dogs (33%) were positives to Leishmania antibodies. 42,6% lived in rural areas and 57,4% in urban areas. However, one of the disadvantages of serological test could be crossreactivity with several agents, like other trypanosomatids (Trypanosoma cruzi and dermotropic leishmania species) and common hemoparasites in dogs, such as Babesia canis and Erlichia canis, emphasizing the need for other confirmatory tests. Ituiutaba is a city of rural habits and is considered an ancient endemic region for Chagas disease, which provides better contact between dogs and countryside, predisposing them to living with wild animals (opossum) and insects such as sand flies, triatomines and ticks. However, due to the expansion process of leishmaniasis in Minas Gerais state and the proximity of the studied area and regions where the presence of L. infantum is already known, the possibility of dogs are already carrying this parasite should not be dismissed, calling attention of the competent authorities for a possible Leishmaniasis establishment in the region.

- EPIDEMIOLOGY [859] P 808 - USE OF GEOTECHNOLOGIES TO ANALYZE THE INCIDENCE OF VISCERAL LEISHMANIASIS IN CAMPO GRANDE, MATO GROSSO DO SUL, BRAZIL, FROM 2007 TO 2011 MORAES, V.S.S.1; GALHARDO, J.A.2; CILLI, M.L.C.1; PARANHOS-FILHO, A.C.1; ANTONIALLI, S.A.C.3 1.LABORATÓRIO DE GEOPROCESSAMENTO PARA ANÁLISES AMBIENTAIS, UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL; 2.FACULDADE DE MEDICINA VETERINÁRIA E ZOOTECNIA, UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL; 3.LABORATÓRIO CENTRAL DE SAÚDE PÚBLICA, SECRETARIA ESTADUAL DE SAÚDE DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL.

Keyword:spatial analysis; geographic information system; incidence

Abstract: Since 2002 Campo Grande, Mato Grosso do Sul, Brazil, has entered the cycle of urbanization of american visceral leishmaniasis (AVL) and canine leishmaniasis. Campo Grande has approximately 800,000 inhabitants and is divided into 74 districts. Aiming to improve the control strategies of the disease, the analysis of the AVL incidence in the districts of Campo Grande in the years 2007 to 2011 was performed using spatial analysis tools. For the the indicator construction were used absolute incidence data from the State Health Department of Mato Grosso do Sul and the population data of the years 2007 and 2010 from Brazilian Institute of Geography and Statistics. The population of the years 2008, 2009 and 2011 was estimated by the average annual geometric growth and descriptive analyzes were performed in Prism 5.0. To determine the spatial relationship between districts and vulnerability areas for AVL a geographic information system using the relative incidences per district / year was built and the Local Moran Index (LMI) calculated. Spatial analysis was performed using ArcGIS 9.3 and thematic maps were developed in Macromedia Freehand 9.0 and MAPublisher 4.0. From 2007 to 2011, 14.9% of the districts had no cases, 85.14% had at least one reported case and 24.32% of the districts were considered critical areas, in which 55.1% (351/630) of cases in the entire period occurred. The average incidence was 126 cases / year, with a slight decrease in the incidence observed in the years 2008 and 2009 followed by a progressive increase in 2010 and 2011. The average incidence /10.000 inhabitants / district ranged from 2.14 in 2009 to 3.87 in 2011. In the spatial analysis, districts with high annual incidence occurred mostly in the south and in 2008, a focus of high incidence in the north and another in the central region were identified. Through LMI, some vulnerability areas have been identified and, over the years, an increase in the number of cases in these areas. The increasing tendency of cases / year and detection of disease in areas of vulnerability that over time has become endemic suggest that the

control measures taken are not effective in controlling the disease at study period. The temporal analysis of the incidence of AVL in Campo Grande using descriptive methods associated with GIS is feasible and constitutes a tool available to the government for planning actions to control disease in our reality.

- EPIDEMIOLOGY [861] P 809 - PREVALENCE OF ANTI-LEISHMANIA INFANTUM CHAGASI ANTIBODIES IN HUMANS AND DOGS FROM TAPIRAPÉ INDIGENOUS COMMUNITY IN MATO GROSSO, BRAZIL LIMA, J.T.R.1; SOARES, H.S.1; MINERVINO, A.H.H.2; MARCILI, A.1; MALHEIROS, A.F.3; SOARES, R.M.1; GENNARI, S.M.1 1.USP, SÃO PAULO, SP, BRAZIL; 2.UFOPA, SANTAREM, PA, BRAZIL; 3.UNEMAT, CÁCERES, MT, BRAZIL.

Keyword:leishmaniasis; dogs; seroprevalence

Abstract: Visceral leishmaniasis is a zoonosis with a cosmopolitan distribution which can infect humans and a wide range of animal species. In Brazil Leishmania infantum chagasi is the etiologic agent of the disease and dogs are the main domestic reservoir. Due to the narrow relation of dogs and men, this specie is the major source of infection to humans. The present study aimed to determine the prevalence of anti-L. infantum chagasi antibodies in humans from Tapirapé indigenous community, and in dogs from the same villages. Samples were collected in seven villages located in Confresa municipality, in Mato Grosso state, Brazilian Amazon from September 2010 to May 2011, totalizing 472 humans and 110 dogs. For the detection of anti-L. infantum chagasi antibodies, the serum samples were tested by Indirect Fluorescence Antibody Test (IFAT 40). The effects of gender, age and hunting activities were analyzed using Fisher’s exact test and P 0.05 were considered significant. All humans tested negative and only two (1.81%) of 110 dogs were positive for anti-L. infantum chagasi antibodies. Both positive dogs were adult males and used to participate of hunting activities. Association among risk factors could not be observed due to the low prevalence observed. The results indicate low distribution of the parasite among humans and dogs in the studied area.

- EPIDEMIOLOGY [867] P 810 - DETECTION OF LEISHMANIA (VIANNIA) SP DNA IN TICKS FROM DOGS IN THE URBAN AREA OF IMPERATRIZ-MA, BRAZIL FREITAS, M.S.1; SILVA, P.H.S.E.2; LEITE, S.T.V.C.3; PANTOJA, K.M.B.3; SANTOS, Y.C.V.3; ISHIKAWA, E.A.Y.1 1.UNIVERSIDADE FEDERAL DO PARÁ, BELÉM, PA, BRAZIL; 2.CENTRO DE CONTROLE DE ZOONOSES, IMPERATRIZ, MA, BRAZIL; 3.ESCOLA SUPERIOR DA AMAZÔNIA, BELÉM, PA, BRAZIL.

Keyword:l. (viannia) sp; ticks; r. sanguineus

Abstract: Detection of infected animals and correct identification of the etiologic agent is crucial for the epidemiological study of leishmaniasis in endemic areas. Among the domestic animals, dogs are important hosts within the transmission cycle. According to the epidemiological surveillance service in Brazil, the municipality of Imperatriz located at 550 km from São Luiz, the capital of the state of Maranhão, is endemic for cutaneous and visceral leishmaniasis. However, research in dogs is still restricted to clinical diagnosis, so we do not know the exact distribution of infected dogs in the city. In this study, we investigated the presence of Leishmania sp in ticks collected from dogs of the petshop and Zoonosis Control Center of Imperatriz using the PCR technique. The DNA was extracted from 640 female of ticks, individually, and tested using

primer that amplified the Leishmania sp mini-exon gene. Ticks were collected from 41 dogs from different neighborhoods of the urban area of Imperatriz. Most ticks were identified as Rhipicephalus sanguineus. The following clinical signs suggestive of leishmaniasis were observed in dogs: onychogryphosis in 53.65% (22/41); ulcers in 63.41% (26/41), hair loss and inappetence in 39.02% (16/41). A percentage of 26.56% of ticks (170/640) collected from 16 (39.02%) dogs had DNA of the Leishmania subgenus Viannia, which causes the cutaneous form of the disease, amplifying a fragment of approximately 250 bp. No DNA of Leishmania infantum was detected. Infected ticks were detected from both symptomatic and asymptomatic dogs. Although it has not been proven that ticks can transmit Leishmania to dogs under natural conditions the result of this study has several important aspects because it is a noninvasive method of detection, able to differentiate groups of circulating parasites, especially if the animals do not have lesions, may be a biological indicator in locations where it is not done the serodiagnosis and entomological survey and can support the local health surveillance programs.

- EPIDEMIOLOGY [882] P 811 - ANALYSIS OF THE EPIDEMIOLOGICAL PROFILE CASES OF AMERICAN CUTANEOUS LEISHMANIASIS IN THE CITY OF PORTO VELHO, RONDONIA, IN THE PERIOD FROM 2007 TO 2011 BAIﾃグ, A.1; FERNANDES, L.S.1; AZEVEDO, F.S.1; GADELHA, S.M.S.2; JUNIOR, C.F.S.1 1.FEDERAL UNIVERSITY OF RONDONIA, PORTO VELHO, RO, BRAZIL; 2.APARICIO CARVALHO INTEGRATED COLLEGE, PORTO VELHO, RO, BRAZIL.

Keyword:epidemiology; american cutaneous leishmaniasis; porto velho

Abstract: This study is a retrospective observational descriptive quantitative one, which described the epidemiological profile cases of American Cutaneous Leishmaniasis (ACL) residing in the city of Porto Velho in the period from 2007 to 2011. The ACL is a disease which is expanding geographically and configures itself as one of the most significant dermatologic diseases in tropical regions of Brazil and the world. It is characterized by being a non-contagious and infectious disease caused by protozoa of the genus Leishmania and transmitted to humans by the order Diptera mosquitoes. It is considered an occupational disease that often affects workers already encroaching on rainforests or live in areas close to them. The study used the database of the Municipal Health Department of Porto Velho - RO, through the Information System for Notifiable Diseases (SINANNet-Leishmaniasis). In the period from 2007 to 2011 were reported 455 cases of Leishmaniasis in the municipality of Porto Velho; of these, 93.2% (424) are new cases and 6.2% (28) are recurrences. The male profile was higher, with 87.7% (399) of notifications. The most affected age group was the 20 to 39 years, with 64.2% (292). As for the area of residence, urban was higher with 57% (259). The most common occupation of those affected by the disease was a farmer / agricultural, with 29.2% and 60% claimed to be work related disease. The cutaneous clinical form was the most affected, with 85.5% (389) and the confirmation criterion more used was the clinical laboratory, with 91.4% (416). Of the total of cases, 69.4% (316) were autochthonous, 17.4% (79) were imported from other locations and 13.2% (60) is of undetermined origin. As for evolution, 84.2% (383) of the cases were cured, 1.1% (5) abandoned treatment and in 13.8% (63) of the cases there is no known developments since the notification was scheduled to option 'ignored / white'. It can conclude from this study that ACL continues shaping up as a very important disease in Porto Velho and throughout the State of Rondonia, with constant and significant incidence in recent years. Since that deserves greater attention from public health authorities in order to minimize the risk factors existing by the geographical and social conditions of population of the region in question.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [885] P 812 - VISCERAL LEISHMANIASIS IN URBAN AREAS: STUDY OF ENTOMOLOGICAL ASPECTS AND CONTROL STRATEGIES SARAIVA, L.1; PEREIRA, A.A.S.1; LEITE, C.G.2; SERRA E MEIRA, P.C.L.1; ABREU, B.L.1; GONTIJO, C.M.F.1; ANDRADE FILHO, J.D.1 1.CENTRO DE PESQUISAS RENÉ RACHOU, BELO HORIZONTE, MG, BRAZIL; 2.PREFEITURA MUNICIPAL DE BELO HORIZONTE, BELO HORIZONTE, MG, BRAZIL.

Keyword:visceral leishmaniais; lutzomyia longipalpis; belo horizonte city

Abstract: The control measures of visceral leishmaniasis (VL) are inefficient in Brazil, even in municipalities with an extensive program of seropositive dogs elimination and insecticide spraying in areas with high incidence of human and canine cases, as occurs in the Belo Horizonte municipality. This is one of the Brazilian municipalities that have higher rates of incidence and mortality by VL. From 1994 to 2011, 1401 cases of human disease were confirmed. The Venda Nova Sanitary District (DS), selected for this study, presents high rates of human cases and canine seropositivity. In 2011, 13 human cases were recorded and the canine seropositivity was 6.5%. The aims of this work are: study the seasonal variation of sand flies in Venda Nova DS; evaluate the impacts of control measures in phlebotomine fauna; determine the natural infection rate of females collected in the area; perform spatial analysis of the variables included in the study and develop entomological indicators based on data investigated. The project was implemented in August 2011 and the filed collections and the investigation of natural infection were initiated. The insects are identified according to Galati 2003. The collected females are submitted to PCR and sequencing reactions for the investigation of natural infection with Leishmania species. From August 2011 to August 2012, 1690 sand flies specimens belonging to six species were collected: Lutzomyia longipalpis, Evandromyia sallesi, Evandromyia cortelezzii, Evandromyia lenti, Nyssomyia whitmani and Micropigomyia schreiberi. The species Lutzomyia longipalpis accounted for 97.28% of the total collected sand flies and was recorded in 15 out of the 17 collection points. Among the total sand flies collected 17.75% (300 individuals) were females and 279 (93%) of these belonged to the species Lu. longipalpis. Fifteen female specimens showed positive PCR reaction for the fragment ITS-1 indicating the infection by trypanosomatid. The positive samples will be identified at the species level through the fragment sequencing. Preliminary analysis of the seasonal variation of sand flies and the curve of insecticide spraying indicates that there is a negative correlation between these two variables. The collections are expected to finish in August 2013 and in the end of this period we will analyze the distribution of all variables together, considering the space and time.

- CONTROL PROGRAMS [894] P 813 - CANINE SEROPREVALENCE OF VISCERAL LEISHMANIASIS IN CEARÁ STATE, BRAZIL IN 2012 CUNHA, J.C.L.1; LIMA, J.W.O.2; VERDIANO, V.A.1; CARIOCA, S.M.1; SOUSA, A.Q.1; COSTA, J.R.A.3; SANTOS, J.L.B.1; MOURA, F.B.P.1 1.SECRETARIA DE SAÚDE DO ESTADO DO CEARÁ, FORTALEZA, CE, BRAZIL; 2.UNIVERSIDADE ESTADUAL DO CEARÁ, FORTALEZA, CE, BRAZIL; 3.SECRETARIA DE VIGILÂNCIA EM SAÚDE, FORTALEZA, CE, BRAZIL.

Keyword:program control; leishmaniasis visceral; canine

Abstract: Due to Calazar urbanization in the last century were created favorable epidemiological conditions for remaining of this disease with high population densities of reservoirs, vectors and human being becoming big cities vulnerable. Dog is the main domestic reservoir of L. chagasi being responsible for maintenance of this parasite in the environment. The effective control of

this disease requires an incidence fall, which works as reservoirs of disease in human. The objective of this present study was to quantify the percentual of positive dogs identified by Leishmaniasis National Program (LNP) in Ceará State in 2012. This is a descriptive and transversal where datum were obtained from reservoirs control reports sent every month by municipalities to LNP. Datum were gathered and presented in absolute frequencies way and relative one. In 2012 were done Leishmaniasis serological exams in 273.715 dogs, which 7.451 were positive showing a positive percentual in 2,72 % in Ceará. Monthly frequencies status varied from 2,01 to 9,76 %, being months which had high values December, August, June and July with 9,76%, 3,52%, 3,36% and 3,23%, respectively. These examined animals were from routine control and spontaneous demand actions of population. Therefore, control actions identified a great concentration of canine positivity through months, which precede high incidence rates of this zoonosis in human, clearing the necessity of implementing control actions of this endemic period.

- EPIDEMIOLOGY [895] P 814 - STUDIES ON THE SAND FLY FAUNA (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN WILD AREAS INHABITED BY HOWLER MONKEYS IN THE NORTHEAST OF ARGENTINA MARTINEZ, M.F.1; SANTINI, M.S.2; KOWALEWSKI, M.M.3; BERROZPE, P.2; SALOMON, O.D.1 1.INTITUTO NACIONAL DE MEDICINA TROPICAL, MINISTERIO DE SALUD DE LA NACIÓN, PUERTO IGUAZÚ, ARGENTINA; 2.CENTRO NACIONAL DE DIAGNÓSTICO E INVESTIGACIÓN EN ENDEMOEPIDEMIAS, MINISTERIO DE SALUD DE LA NACIÓN, BUENOS AIRES, ARGENTINA; 3.ESTACIÓN BIOLÓGICA DE CORRIENTES, MUSEO ARGENTINO DE CIENCIAS NATURALES, CORRIENTES, ARGENTINA.

Keyword:wild environment; height traps; phlebotominae

Abstract: In Argentina Leishmania braziliensis has been identified as the main agent causing cutaneous leishmaniasis. Although the reservoir of L. braziliensis has not been identified yet, as no animal meets all the criteria required to be defined as such, horses, cats, canids and rodents are susceptible to infection. In this country, the Phlebotominae sand flies Nyssomyia neivai, Ny. whitmani, and Migonemia migonei have been confirmed as the most frequent vectors of L. braziliensis. In rural houses situated near our study area 7 sand fly species had been recorded: Lutzomyia longipalpis, Ny. neivai, Brumptomyia sp., Mg. migonei, Ny. whitmani, Evandromyia cortelezi-salessi, and Psathyromyia shannoni. The goal of this study is to describe the Phlebotominaes species present in wild areas inhabited by howler monkeys (Alouatta caraya), and asses if community composition of sand flies depends on the height above the ground.

Samples were collected in February 2012 in a locality of northeastern Argentina (27º30’S, 58º41’O). Sand fly traps (CDC-type minitraps) were placed at 10 howler-monkey sleeping sites separated by 1,5km on average, and spanning a total area of 14km2. Traps were hung at 1,5m (low) and 7m (high) above the ground in each sleeping site and set on for two consecutive nights. The specimens were cleared with lacto-phenol and identified with the aid of a microscope. A total of 626 sand flies were collected from the “low” traps, belonging to six species: Ny. neivai (46,2%), Brumptomyia sp. (33,7%), Mg. migonei (19,2%), Ny. whitmani (0,48%), Ev. cortelezi-salessi (0,32%), and Psa. shannoni (0,16%). A total of 252 sand flies were collected from the “high” traps, representing five species: Ny. neivai (41,7%), Mg. migonei (35,3%), Brumptomyia sp. (20,2%), Ny. whitmani (1,6%), and Psa. shannoni (1,2%). The abundance of the different sand fly species was influenced by the height above the ground (X2= 38,35; p-value < 0,0001). All the species of sand flies captured from the wild environment in this study, were also recorded along this region in the domestic environment in a previous survey. No specimen of Lu. longipalpis was obtained in the wild, whereas this species was frequently recorded in domestic habitats in this region. The sand fly females occurring at upper heights may be using howler monkeys as food sources. Therefore, A. caraya might act as a

reservoir host of leishmanias or, at least, as a population amplifier of its vectors in the wild environment.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [898] P 815 - SANDFLY FAUNA DESCRIPTION (DIPTERA: PSYCHODIDAE) IN AN ENDEMIC AREA OF AMERICAN TEGUMENTARY LEISHMANIASIS IN CEARÁ STATE, BRAZIL. CUNHA, J.C.L.1; LIMA, J.W.O.2; DE SOUSA, L.C.1; CUNHA, G.V.D.3; MOREIRA, P.A.C.1; SILVA, L.O.R.1; ARAÚJO, J.M.P.1; SOUSA, F.D.G.3; MOURA, F.B.P.1; SANTOS, F.K.M.1; VERDIANO, V.A.1 1.SECRETARIA DE SAÚDE DO ESTADO DO CEARÁ, FORTALEZA, CE, BRAZIL; 2.UNIVERSIDADE ESTADUAL DO CEARÁ, FORTALEZA, CE, BRAZIL; 3.SECRETARIA DE SAÚDE DO MUNICÍPIO DE VIÇOSA DO CEARÁ, VIÇOSA DO CEARÁ, CE, BRAZIL.

Keyword:vectors; leishmaniasis tegumentary; ecology

Abstract: Sandfly fauna knowledge is indispensable in process understanding of American Tegumentary Leishmaniasis transmission. In Ceará Leishmaniasis have been showed in an endemic way, intercalated by epidemic periods concerning a great importance. This study aimed to describe sandfly fauna in a well known endemic area of Tegumentary Leishmaniasis in Viçosa do Ceará municipality, Chapada do Araripe in Ceará North area. It was done a set up of the entomological survey in four places. Captures were done using light traps placed in a house in each place during three consecutive nights in June 2012. All collected sandflies were marked and identified in Tomás Correia Aragão Medical Entomology Laboratory belonged to Health Secretary of Ceará State. Worked places were: Buíra, Canto da Buíra, Porteiras and Buirinha. In all were captured 1.520 specimens, which 65,7%, 27,8% and 1,8% were L. whitmani, migonei and longipalpis, being 82% surrounding home and 18% inside home, which 67% were males and 33% female. Prevalence order from place and city were similar to general ones. The studied area presents high predominance being these ones the main responsible reservoirs by Tegumentary Leishmaniasis transmission, in addition the presence of L. longipalpis, which could cause an outbreak of Viscera Leishmaniasis in the future.

- EPIDEMIOLOGY [899] P 816 - POOR HOUSING STRUCTURE IS A RISK FACTOR FOR INFECTION BY LEISHMANIA INFANTUM IN A COHORT OF DOGS FROM AN ENDEMIC URBAN AREA COURA-VITAL, W.1; VIANNA, E.N.1; MARQUES, L.H.S.1; ARAÚJO, V.E.M.2; REIS, L.E.S.3; BRAGA, S.L.3; VELOSO, V.M.3; MARQUES, M.J.4; REIS, A.B.3; CARNEIRO, M.1

1.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 2.SECRETARIA MUNICIPAL DE SAÚDE, PREFEITURA DE BELO HORIZONTE, BELO HORIZONTE, MG, BRAZIL; 3.UNIVERSIDADE FEDERAL DE OURO PRETO, OURO PRETO, MG, BRAZIL; 4.UNIVERSIDADE FEDERAL DE ALFENAS, ALFENAS, MG, BRAZIL.

Keyword:canine visceral leishmaniasis; risk factor; cohort study

Abstract: Visceral leishmaniasis (VL) is caused by Leishmania infantum parasites and transmitted by sand flies of the Phlebotominae family. Dogs are the main urban reservoirs and represent the major source of infection for the vector. Studies reported that some dogs showed PCR-positive results months before seroconversion. This is a concurrent cohort study that was designed to identify the incidence and the risk factors to L. infantum infection detected by PCR-RFLP. This study was conducted in Belo Horizonte, the capital of the State of Minas Gerais, located in Southeastern Brazil, which is considered an endemic area of VL. To determine the predictors of risk for infection by L. infantum, a canine survey (n=1443) was carried out, after a cohort study was established (n=282) in which 3 evaluations were carried out for approximately 1.5-year

follow-up period. Molecular methods (PCR-RFLP) and a structured questionnaire were used. To identify the factors associated with infection (PCR-RFLP) Cox regression model was performed. The overall incidence of infection was 5.7/100 dogs-months (95%CI 5.0–6.5).

Infection with L. infantum was associated with a previous case of canine visceral leishmaniasis (CVL) (HR 1.4; 95% CI 1.1–1.8) and house walls unplastered (HR 3.6; 95% CI 1.6–8.1). Our results identified domiciles that were most vulnerable to occurrence of canine infection by L. infantum. The risk factors reported may contribute in the actions of control measures demonstrating the necessity of insecticide spraying in cracks and crevices in the walls of houses, to reduce biting rates within and around them. Furthermore, our results demonstrate that the control program should adopt measures of environmental management in homes where a previous case of CVL has occurred, since it is more vulnerab - EPIDEMIOLOGY [901] P 817 - EVALUATION OF FILLING IN FIELDS ESSENTIAL FOR VISCERAL LEISHMANIASIS NOTIFICATION. STATE OF BAHIA, 2009-2011 SANTOS, R.S.S.S.; CASTRO, J.S.M. SECRETARIA DA SAÚDE DO ESTADO DA BAHIA, SALVADOR, BA, BRAZIL.

Keyword:epidemiologia; leishmaniose viceral; sistema de informação

Abstract: Introduction: Visceral Leishmaniasis (VL) is a vector-borne disease caused by several species of Leishmania. It Is widely distributed throughout the world. In Brazil the measures of prevention and control are guided by the notification of the cases in Information System for Notifiable Diseases (SINAN). The SINAN guides the management of epidemiological surveillance generating information for planning of control measures and to assist in decision making. Data collection is an important step in this process. Filling mistaken or not completing one or more variables may bias the assessment and implementation of necessary interventions harm.

Objectives: To assess the degree of fulfillment of the individual records of notification of Visceral Leishmaniasis in the State of Bahia, in the period from 2009 to 2011. Methods: The study was conducted based on information obtained from the application using SINAN TABWIN. Variables selected from the general identification of cases and research. We excluded the required fields to include the case in the system. Variables with missing and / or ignored were considered incomplete. Results: During the study period 1757 cases were reported as follows: 560 in 2009, 590 in 2010 and 607 in 2011. Regarding the percentage of fill of all selected variables, in each period studied was 79% or more. However, when analyzing the variables recorded individually and stratified by reporting and investigation, ranged from 4 to 100%. Considerations: The results indicate that the management and planning of epidemiological surveillance and control of visceral leshimaniose can be performed based on data available at the Sinan. However, it is necessary to intensify the monitoring of data and continuing education of professionals about the importance of filling full of tools for reporting and investigating effectiveness and timeliness of public health interventions recommended - VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [908] P 818 - DNA BARCODING FOR IDENTIFICATION OF SAND FLIES (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) FROM THE ATLANTIC FOREST OF THE STATE OF ESPÍRITO SANTO, SOUTHEASTERN BRAZIL PINTO, I.S.1; FERREIRA, A.L.2; CHAGAS, B.D.1; ANDRADE-FILHO, J.D.3; GALATI, E.A.B.4; FUZARI, A.A.5; BRAZIL, R.P.5; PEIXOTO, A.A.1 1.LABORATÓRIO DE BIOLOGIA MOLECULAR DE INSETOS, INSTITUTO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL; 2.DEPARTAMENTO DE PATOLOGIA, UNIVERSIDADE FEDERAL DO ESPÍRITO SANTO, VITORIA, ES, BRAZIL; 3.CENTRO DE REFERÊNCIA NACIONAL E INTERNACIONAL PARA FLEBOTOMÍNEOS, CENTRO DE PESQUISAS RENÉ RACHOU, BELO HORIZONTE, MG, BRAZIL; 4.DEPARTAMENTO DE EPIDEMIOLOGIA, FACULDADE DE SAÚDE PÚBLICA DA UNIVERSIDADE

DE SÃO PAULO, SAO PAULO, SP, BRAZIL; 5.LABORATÓRIO DE BIOQUIMICA E FISIOLOGIA DE INSETOS, INSTITUTO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:biodiversity; systematics; coi

Abstract: The insufficient numbers of qualified taxonomists to describe the remaining biodiversity, as well as the great morphological similarity between some sand fly species which are vectors of Leishmania parasites, highlight the necessity to use new species identification tools that are faster than the traditional systematic methods relying on morphology. Currently, the most used of these new methods is DNA barcode. We carried out a preliminary analysis to evaluate the utility of this approach towards species identification of phlebotomine sand flies from the Atlantic Forest in the state of Espírito Santo, Southeastern Brazil. DNA was extracted from legs and used in PCR and sequencing of a COI gene fragment from individual sand flies, while the remaining bodies and wings were mounted in slides for morphological species identification.

We analyzed so far more than 250 sand fly specimens belonging to 25 morphological species and 12 genera as follows: Brumptomyia figueiredoi, Brumptomyia nitzulescui, Evandromyia edwardsi, Evandromyia lenti, Evandromyia tupynambai, Expapillata firmatoi, Lutzomyia longipalpis, Micropygomyia capixaba, Micropygomyia ferreirana, Micropygomyia quinquefer, Micropygomyia schreiberi, Migonemyia migonei, Nyssomyia intermedia, Nyssomyia whitmani, Pintomyia fischeri, Pintomyia misionensis, Pintomyia monticola, Pressatia choti, Psathyromyia lutziana, Psathyromyia pascalei, Psathyromyia pelloni, Psathyromyia shannoni, Psychodopygus hirsutus, Psychodopygus matosi and Sciopemyia microps. Phylogenetic analysis of the sequences that we generated indicates that the observed species clades are congruent with the morphological identification in almost all the cases. All the incongruences were generated by morphological misidentifications, which were solved by reanalyzes of sand fly slides. The results obtained so far show that the fragment of COI gene used in DNA barcoding will be useful for the molecular identification of sand flies although the data should be analyzed carefully due to problems of introgression often associated with this genetic marker.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [912] P 819 - STUDY OF SANDY FLY FAUNA (DIPTERA, PSYCHODIDAE) IN MARANHÃO STATE, BRAZIL DUAILIBE, R.F.1; REBELO, J.M.M.1; MORAES, J.L.P.2; FILHO, A.A.P.3; MELO, M.N.3; SILVA, S.O.3 1.UFMA, SÃO LUIS, MA, BRAZIL; 2.FUNASA, SÃO LUIS, MA, BRAZIL; 3.UFMG, BELO HORIZONTE, MG, BRAZIL.

Keyword:cutaneous leishmaniasis ; visceral leishmaniasis ; vector

Abstract: Sand flies are vectors of protozoan parasites of the genus Leishmania. These insects are widely distributed in tropical forests can be found in many different environments. It's a very diverse group of insects being cataloged about 800 species worldwide, about 480 are distributed throughout the Americas. In Maranhão, has identified 91 species, 19 of these are potentially genus Lutzomyia vectors. Many species are associated with anthropogenic environments narrowing the human-vector. Major human activities have reduced natural areas, threatening biodiversity in areas of occurrence of sandflies. In Maranhão between 2007 and 2010 were 13,192 diagnosed cases of Cutaneous Leishmaniasis and Visceral Leishmaniasis in 2482, a fact that motivated us to perform this work. The aim of this study was to determine the species of sandflies in household environments, peridomicilirares and wild areas studied. The study took place in the towns of Chapadinha, Hills and Caxias in the localities Buritizinho, Bom Jesus and Paul, respectively, in these localities selected two houses, traps them installed HP to 1.5 m tall one inside and around the homes and the other in the woods. Three consecutive days of

sampling once a month for two years. From May to August 2012, 11,600 specimens were found in 9448 were Colinas sandflies belonging to 10 species in 1083 Chapadinha 17 and Caxias 1069 15. Were found in the study sites considered species of sandfly vectors of leishmaniasis effective, among them are: L. longipalpis, L. complex, L. flaviscutellata, L. migonei, L. umbratilis, L. wellcomei, L. whitmani among others. L. whitmani was the most abundant, present in all locations. This represents an important aspect of this species is recognized as a vector of TL. L. longipalpis was second in abundance and corresponds to one of the main vectors of VL in these municipalities. Thus, this study demonstrated the existence of a sand flies with the presence of diverse species considered vectors of Leishmania, which indicates the need for vector control measures in the areas under study in order to decrease the incidence of VL and TL these locations.

- RESERVOIRS [915] P 820 - PREVALENCE RATE OF CANINE VISCERAL LEISHMANIASIS AND DENSITY OF LUTZOMYIA LONGIPALPIS (LUTZ & NEIVA, 1912) IN BARRA DO GARÇAS, STATE OF MATO GROSSO, BRAZIL MORAES, S.C.1; QUEIROZ, M.F.M.2; VARJÃO, J.R.2 1.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 2.SECRETARIA DE ESTADO DE SAÚDE DE MATO GROSSO, BARRA DO GARÇAS, MT, BRAZIL.

Keyword:canine visceral leishmaniasis; lutzomyia longipalpis; reservoirs

Abstract: The visceral leishmaniasis (VL) is a severe disease is caused by the protozoan, Leishmania infantum (syn. L. chagasi) in Latin America. It is a zoonosis in wich the domestic dog is the main reservoir in urban and periurban transmission. The parasite is mainly transmitted by sand fly Lutzomyia longipalpis, vector adapted to the peridomestic environment and widely distributed in Brazil. The objective of the study was to analyze the relationship of the density of L. longipalpis in the transmission of canine visceral leishmaniasis (CVL) over the years in three districts of the municipality of Barra do Garças, State of Mato Grosso, Brazil with reported cases of human visceral leishmaniasis. From September 2004 to August 2006 was conducted entomological monitoring with CDC light traps were installed in peridomestic locations. We calculated the relative frequency and richness of sandflies. Information from canine serological inquiry was carried out in 2004 to 2011 were collected from the Municipal Health Barra do Garças, with these data we calculated the prevalence rate of LV canina. The sandfly population was composed of 2 genera and 27 species, totaling 8,097 individuals collected. The most frequent was L. longipalpis (44%), followed by L. lenti (18.9%), L. whitmani (13.9), L. carmelinoi (9.1%), L. evandroi (5.1%), L. termitophila (3.3%), L. sordellii (1.9%) and twenty other species (less than 4%). The index of species richness (D ) was 6.65, indicate high diversity. The predominant species was L. longipalpis with relative frequency presented at each location, Novo Horizonte (75.18), São José (19.75) and Vila Maria (5.07). The prevalence rate of LVC showed the following results: Novo Horizonte (3.68, 6.84, 1.05, 0.53, 9.87, 23.61, 13.73 and 18.08), São José (0.80, 0.64, 0.0, 0.96, 3.72, 0.0, 2.14 and 11:34) and Vila Maria (0.38, 0.0, 0.0, 0.57, 3.33, 0.0, 5.87, 4.26) respectively for each year analyzed. The district with the highest population density of L. longipalpis presented over the years highest prevalence rate in dogs.

The high presence of vectors and the high prevalence rate of LV canine increases the risk of transmission of the human LV. The monitoring and evaluation of activities undertaken by municipalities for the LV should guide local planning activities for the protection and promotion of Health to reduce the morbidity and mortality related to LV. - VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [917]

P 821 - EVALUATION OF CONTROL MEASURES OF VISCERAL LEISHMANIASIS IN BELO HORIZONTE, BRAZIL, BETWEEN 2010 AND 2012.

SAID, R.F.D.C.1; FIUZA, V.O.1; PESSANHA, J.E.M.1; ALMEIDA, M.C.M.1; BRANDÃO, S.T.1; DIAS, E.S.2 1.SECRETARIA MUNICIPAL DE SAUDE, BELO HORIZONTE, MG, BRAZIL; 2.CPQRR, BELO HORIZONTE, MG, BRAZIL.

Keyword:visceral leishmaniasis; control measures; belo horizonte

Abstract: The visceral leishmaniasis (VL) is widely distributed in Belo Horizonte, the capital of the Brazilian state of Minas Gerais. A cross-sectional study was performed in order to evaluate some variables involved in VL dynamics to subside adequate interventions. Our study was developed in two regions (B and O) of Belo Horizonte based on VL transmission profiles and occupation characteristics (3,182 and 2,887 households, respectively). The study was divided in two periods of 12 months each: October 2010 to September 2011; October 2011 to September 2012. Both regions were divided in 3 sectors based on the type of intervention applied: controlcanine surveys and removal of seropositive dogs; spraying- canine surveys, removal of seropositive dogs and residual chemical control; environmental management- canine surveys, removal of seropositive dogs and environmental management. The sanitary conditions of the households were recorded and the population of Lutzomyia longipalpis was monthly monitored using HP light traps. There were no reports of human VL cases during the study. Three canine surveys were performed with 2,009 samples collected in B and 2,319 collected in O. In B, 100 dogs were seropositive for VL and 93 (93%) of them were euthanized. In O, the number of seropositive dogs decreased to 87 with euthanasia of 62 (71%) of them. The average canine seroprevalence per year was 5.26%, 2.5% and 4.0% in the O region; 8.7%, 3.9% and 2.5% in the B region. Concerning the sectors, the average prevalence rates ranged from 0.8% to 11.3 %, with significant reduction in the spraying and control sectors in B. A total of 253 and 4,846 specimens of L. longipalpis were captured in the B and in the O regions, respectively. In both, the vector population decreased during the second year of study; however there was no significant difference in the results among the sectors. A total of 2,191 (66%) and 2,527 (94%) households were sprayed in B and O, respectively, in three cycles. Twenty-two and 28 tons of solid waste were removed from the environmental management sections in B and in O, respectively. Our results suggest a decrease in the vector population and in the canine prevalence in the second year of the study. However, a more detailed analysis will be needed considering the limitations and the complexity of the control actions in urban areas.

Financial Support: FAPEMIG, FIOCRUZ, Secretaria Municipal de Saúde de Belo Horizonte, CNPq.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [918] P 822 - PERMETHRIN-IMPREGNATED MOSQUITO-NETS EFFECTIVENESS TESTED ON EXPERIMENTAL HENHOUSES: PRELIMINAR STUDY FOR PREVENTION OF CUTANEOUS LEISHMANIASIS IN NE ARGENTINA SALOMON, O.D.; MANTECA ACOSTA, M.; MOLINA, N. INSTITUTO NACIONAL DE MEDICINA TROPICAL, PUERTO IGUAZÚ, ARGENTINA.

Keyword:curtains; insecticide-impregnated; henhouse

Abstract:

The use of insecticide-impregnated fabrics as preventive tools has become a relevant topic for research and control of vector borne diseases due to its effect decreasing vector-human contact, and the possibility to be applied at the local level with the active participation of communities.

In this work we used non-commercial curtains since our objective is to find a participative control strategy where the own community can make and use the curtains. We impregnated them with permethrin because of its moderate volatility and toxicity in mammals, and for its high insecticide and repellent activity. The project is being developed in a rural area of Puerto Iguazú (Misiones, Argentina) where epidemic outbreaks of Cutaneous Leishmaniasis were reported in 2004-2005 with Nyssomya whitmani as incriminated vector. The objective of this work was to evaluate the changes of phlebotomine abundance in presence of 10% permethrineimpregnated mosquito-nets in semi-field conditions. We installed three experimental henhouses as phlebotomine capture stations in the border of the forest, leaving 50 meters between them.

The treatment (impregnated curtain, IC) and controls (non-impregnated curtain, NIC; without curtain, WC) were assigned randomly. To simulate incomplete obstruction conditions of houses, curtains were set as to cover only two sides of the henhouse. During two consecutive nights, a light trap was set from 17 pm to 9 am to measure phlebotomine abundance. This design was repeated once per month from February 2012 to January 2013. In this preliminary analysis we used data from November, December and Februaryanalyzed with an ANOVA including “night” as blocking factor. Autumn and winter months (April-June) were excluded because of low abundances registered in past studies.In the warm months sampled up to now we observed a significant difference in phlebotomine/trap/night abundance between treatments (F2,30=13.15, p=0.0001). Abundance in IC henhouses (25.65, SD 39.29) was lower than NIC (238.82, SD 476.21) and WC (453.43, SD 578.13). In these months, abundance decreased 89.26% in presence of an impregnated curtain compared to abundance in absence of curtain. In the remainig months (March, August, September, October) we found a similar trend but with greater variance of mean abundance within treatments. In the future we expect to build more complex models so as to evaluate the effectiveness of treatments considering daily and annual variation in phlebotomine abundance.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [926] P 823 - THE SAND FLY FAUNA (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN CORUMBÁ, MATO GROSSO DO SUL, BRAZIL. PRELIMINARY RESULTS. CASARIL, A.E.1; DE OLIVEIRA, E.F.2; RIBEIRO, L.M.1; IGUCHI, G.U.1; MONACO, N.Z.3; SERRA, Q.D.3; OSHIRO, E.T.1; INFRAN, J.M.1; DE FIGUEIREDO, H.R.1; OLIVEIRA, A.G.1 1.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL; 2.FACULDADE DE SAÚDE PÚBLICA DA UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 3.CENTRO DE CONTROLE DE ZOONOSE DE CORUMBÁ, CORUMBÁ, MS, BRAZIL.

Keyword:phlebotomine; corumbá; fauna

Abstract: The municipality of Corumbá is considered area of intensive transmission for visceral leishmaniasis (VL) and reported 200 cases of the disease in the period from 1999 to 2011 and seven confirmed cases of American cutaneous leishmaniasis (ACL) from 2007 to 2011. Fauna studies leading up to the association between the occurrence of these insects and the prevalence of the leishmaniasis in the areas with records of the diseases and promoting measures for the control and surveillance. The aim of this study was to identify the characteristics of sand flies (Diptera: Psychodidae: Phlebotominae) in the Municipality of Corumbá, Mato Grosso do Sul, Brazil. The Sand flies were captured weekly with light automatic traps at 06:00 pm to 07:00 am, in peri and intradomicile environments in five urban districts, from April to September 2012.

The sand flies were sent to the Laboratory of Parasitology at the Federal University of Mato Grosso do Sul. They were clarified and identified following the classification proposed by Galati (2003). The 115 weekly captures yielded 4,549 specimens of phlebotomine, 3,877 males

(85.22%) and 672 females (14.78%). The fauna was composed of 11 species: Brumptomyia brumpti, Evandromyia cortelezzii, Evandromyia corumbaensis, Evandromyia sallesi, Evandromyia walkeri, Lutzomyia cruzi, Lutzomyia forattinii, Micropygomyia peresi, Martinsmyia oliveirai, Psathyromyia shannoni and Sciopemyia sordellii. The most abundant species was Lu. cruzi representing 93.65% and the second was Lu. forattinii with 3.82%.

Overall, the insects were more frequently captured in peridomicile (64.65%) than in the intradomicile (35.35%) of the residences. This study is being conducted for 24 months. Financial support: FAPESP. - CONTROL PROGRAMS [933] P 824 - CHALLENGES FOR UNDERSTANDING ECO-EPIDEMIOLOGICAL PROCESSES IN THE INSTALLATION OF VISCERAL LEISHMANIASIS MARZOCHI, M.C.A.1; MARZOCHI, K.B.F.1; FAGUNDES, A.1; DE SOUZA, M.B.2; ANDRADE, M.V.3; MADEIRA, M.F.1; MOUTA-CONFORT, E.1; SCHUBACH, A.O.1

1.INSTITUTO DE PESQUISA CLINICA EVANDRO CHAGAS-FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 2.ESCOLA NACIONAL DE SAÚDE PÚBLICA SERGIO AROUCA-FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.SECRETARIA MUNICIPAL DE SAÚDE E DEFESA CIVIL, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:visceral leishmaniasis; eco-epidemiology; control

Abstract: From 1977 (index case) to 2006, 87 cases of visceral leishmaniasis (VL) were confirmed in Rio de Janeiro, Brazil, with isolation of Leishmania chagasi in humans and dogs in peri-urban areas in the North and West Zones of the municipality, in foci with extremely poor human dwellings, varying amounts and types of vegetation, and the constant presence of dogs and sand flies. The first VL cases were male adults; the majority of all cases were over 5 years of age (65%) and males (61.5%). Case-fatality (10.4%) predominated in individuals less than 14 years of age, with no difference by gender. There were 2 cases of VL/HIV co-infection. An association was assumed between emergence of the disease and human and canine migrations with disorderly occupation of the territory, deforestation, and poverty. Steady reduction of VL (no further human cases) is attributed to 30 years of close epidemiological surveillance in the risk areas, with active case searches and an emphasis on education, mainly cleaning the yards around the human dwellings, combined with vector control using insecticides (organochlorines until 1991, then organophosphates and pyrethroids) and culling seropositive dogs. VL dropped from 2.8 human cases per 100,000 inhabitants in 1981 to less than 0.01/100,000 in 1997, and no cases have been reported since 2007; canine infection dropped from 4.6% in 1984 to 1.62% in 2008.

Many areas in Rio de Janeiro still suffer the same conditions as those prevailing where VL developed in the 1980s, but have never been affected by the disease. Meanwhile, in various other regions of Brazil where the vector was also previously absent, VL foci have continued to emerge in properly urbanized areas with good housing.

We thus call attention to: the installation of canine and/or human VL in Brazil with variable ecoepidemiological patterns in terms of environmental, social, and economic components; lack of knowledge concerning the variables sustaining the emergence of VL with such clearly differentiated patterns; and the priority of investigating processes involved in the installation, adaptation, and persistence of the most important VL vector in Brazil, Lutzomyia longipalpis, aimed mainly at monitoring and preventing its introduction. (We also recall the possibilities for passive transfer of the vector’s eggs, larvae, and pupae in organic residues such as cereals and animal feces, through railway and highway transportation.)

- EPIDEMIOLOGY [938] P 825 - DIAGNOSIS OF URBAN VISCERAL LEISHMANIASIS IN PUERTO IGUAZÚ CITY, MISIONES PROVINCE, ARGENTINA GÓMEZ, A.1; DESCHUTTER, J.2; GALARZA, F.2; SILVA, G.2; SALOMÓN, O.3; SANTINI, S.4; RAMOS, M.5; FATTORE, G.1; BÁEZ, P.1; ABRIL, M.1 1.FUNDACIÓN MUNDO SANO, BUENOS AIRES, ARGENTINA; 2.MINISTERIO DE SALUD DE LA PROVINCIA DE MISIONES, POSADAS, ARGENTINA; 3.INSTITUTO NACIONAL DE MEDICINA TROPICAL, PUERTO IGUAZÚ, ARGENTINA; 4.CENTRO NACIONAL DE DIAGNÓTIGO E INVESTIGACIÓN EN ENDEMO-EPIDEMIAS, BUENOS AIRES, ARGENTINA; 5.DEPARTAMENTO DE ZOONOSIS MUNICIPAL, PUERTO IGUAZÚ, ARGENTINA.

Keyword:visceral leishmaniasis; lutzomyia longipalpis; transmission risk

Abstract: Puerto Iguazú is a city in the northeast of Argentina, located in Misiones Province, in the zone called Triple Border Area; its neighbor cities are Ciudad del Este, Paraguay, and Foz do Iguacu, Brazil. Based on the regular recording of cases of urban visceral leishmaniasis in Brazil and Paraguay, and the presence of cases in humans and dogs with visceral symptomatology in Argentina, in 2011, this project was started with the aim to conduct an integral study of the visceral leishmaniasis situation in this city, and to deliver a first diagnosis of this health problem. The project includes two components: 1. The entomological aspect, to determine the distribution and dispersion of Lutzomyia longipalpis, the main vector of visceral leishmaniasis in the Americas. For that purpose, four captures would be taken spanning an annual period, one at each change of seasons, taking areas of 400 x 400 meters as sampling units. CDC mini light traps were used, with a sampling effort of two consecutive nights per point (51 points in total) for each capture; 2. Local dog population and its role as reservoir. Samples of blood and lymphatic ganglia were took from 350 dogs randomly selected. These samples were subsequently evaluated for diagnosis of visceral leishmaniasis using the following techniques: Immunochromatography (RK39), Indirect Immunofluorescence (IFI) and Polymerase Chain Reaction (PCR). It was observed that Lu. longipalpis was present throughout the year, being more abundant in the summer-fall time of the year (2831 phlebotomus) reported in 67% of households, and less abundant at the start of the winter, (63 phlebotomus) reported in 10% of households. As regards of the dogs, until now 209 samples have been studied, seven of them tested positive for the three tests, and 11 tested positive for two of the three tests. Results for the remaining 184 dogs were negative on the three tests. These results signal a certain state of visceral leishmaniasis transmission risk in Puerto Iguazú, due to the coexistence of the vector disease and the dogs that act as reservoirs in a continuous way throughout the year. Aware of this situation, we plans to go ahead with the design and implementation of local integrated management actions, including monitoring the vector, the evaluation of suspicious dogs, and deploying a control strategy to prevent the transmission of visceral leishmaniasis to the community.

- EPIDEMIOLOGY [940] P 826 - OCCURRENCE OF ANTI-LEISHMANIA SPP. ANTIBODIES IN DOMICILED DOGS FROM THE CITY OF FOZ DO IGUAÇU, STATE OF PARANÁ, BRAZIL. DIAS, R.C.F.1; SOCCOL, V.T.; BISETTO JÚNIOR, A.2; POZZOLO, E.M.3; CHIYO, L.4; FREIRE, R.L.1; MITSUKA BREGANÓ, R.5; PASQUALI, A.K.S.1; ALBAN, S.6; FENDRICH, R.C.6; CALDART, E.T.1; NAVARRO, I.T.1 1.DEPARTAMENTO DE MEDICINA VETERINÁRIA PREVENTIVA, UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL; 2.SECRETARIA DE SAÚDE DO ESTADO DO PARANÁ, CURITIBA, PR, BRAZIL; 3.19ª REGIONAL DE SAÚDE DO ESTADO DO PARANÁ, FOZ DO IGUAÇU, PR, BRAZIL; 4.CENTRO DE CONTROLE DE ZOONOSES, FOZ DO IGUAÇU, PR, BRAZIL; 5.DEPARTAMENTO DE CIÊNCIAS PATOLÓGICAS, UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL; 6.PROGRAMA DE PÓS-GRADUAÇÃO EM ENG. DE BIOPROCESSOS E BIOTECNOLOGIA, UNIVERSIDADE FEDERAL DO PARANÁ, CURITIBA, PR, BRAZIL.

Keyword:visceral leishmaniasis; leishmania infantum; pcr

Abstract: Visceral Leishmaniasis (VL) is the most severe form of Leishmaniasis with occurrence in Brazil. It is caused by the protozoa Leishmania (Leishmania) infantum, which is generally transmitted by infected female of Lutzomyia longipalpis. VL is endemic in some Brazilian regions with high incidence in urban and peri-urban areas. From the Brazilian southern region, two of the three states (Rio Grande do Sul and Santa Catarina) have reported autochthonous cases. In the other state from that region (Paraná), Lu. longipalpis was first identified in March 2012, in the city of Foz do Iguaçu. The entomological search was performed in 61 residences of six different urban areas, where 40 specimens of Lu. longipalpis were gathered. Using parasitological, serological and molecular methods, it was investigated the presence of L. infantum in blood samples collected from dogs living in residences near the traps used in the entomological research, from the 14th to the 18th of May 2012. From the 350 analyzed samples, 11 (3.1%) were positive by Indirect Immunofluorescence Assay – IFA (cutoff 1:40, being that: 3 were performed with title 40, 3 with 80, 1 with 160, and 4 with 320); and 22 (6.3%) were positive by Rapid Immunochromatographic Test Dpp® Canine Visceral Leishmaniasis and 168 (48.3%) were positive by Enzyme-Linked Immunosorbent Assay - ELISA (cutoff 0.195).

Parasites were searched in 30 dogs that presented two or three positive serological results, in their blood, lymph nodes and DNA by Polymerase Chain Reaction (PCR). Five culture strains were isolated, and 18 animals presented DNA amplified with specific primer (RV1/RV2) to L. (L.) infantum. The conditions that are most likely to explain these results and the agent dispersion are the environmental and climate changes, closeness to endemic regions, road and air axis, and great migration flows allied to the presence of the vector Lu. longipalpis.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [942] P 827 - NATURAL BREEDING PLACES FOR LUTZOMYIA LONGIPALPIS IN AN AREA ENDEMIC FOR VISCERAL LEISHMANIASIS IN BAHIA STATE, BRAZIL OLIVEIRA, D.F. CENTRO DE PESQUISAS GONÇALO MONIZ - FIOCRUZ BAHIA, SALVADOR, BA, BRAZIL.

Keyword:natural breeding; immature form, vector,; weather.

Abstract: INTRODUCTION: Until recently, few micro-habitat were identified as breeding places of sandflies, and most successful work in humid climates. OBJECTIVE: The present study was conducted in a semiarid climate region, investigated the occurrence of natural breeding sites in a series of micro-habitats selected according to the presence of moisture and organic matter, besides reduction of luminosity. METHODS: 64 soil samples were monthly collected in peridomestic environment and household of five homes located in the district of Cavunge Ipecaetá (Bahia-Brazil) were analyzed by direct observation techniques and flotation.

Entomological captures were also performed, providing monthly light traps in peridomestic and indoor environments. RESULTS: Of the 1523 soil samples collected, 49 were positive, and we found a total of 64 sand flies, the majority (n = 40) belonging to the species L. longipalpis.

Similarly, phlebotomines captured between 2002 and identified in the majority (n = 1907) belong to the species L. longipalpis. The relationships between climatic variables and the emergence of sandflies and sandflies captured, there was a relationship of moderate strength and statistically significant between the week before the capture entomological and density of sandflies. indicating that for a region that semiarid climatic parameter is more appropriate in predicting the amount of adult forms of sandflies. CONCLUSIONS: Taken together, our results concerning the characterization of both natural breeding sites of sandflies and their relation to climatic variables, can serve as an auxiliary tool in the direction of biological control.

- EPIDEMIOLOGY [944] P 828 - PREVALENCE OF VISCERAL LEISHMANIASIS IN PALMAS, TOCANTINS PROVINCE, BRAZIL GALVÃO, J.D.1; PERIM, M.C.1; SILVA, K.B.2; PINHEIRO, S.B.1; AVERSI-FERREIRA, T.A.1 1.UNIVERSIDADE FEDERAL DO TOCANTINS, PALMAS, TO, BRAZIL; 2.SECRETARIA MUNICIPAL DE SAÚDE, PALMAS, TO, BRAZIL.

Keyword:prevalence, incidence; visceral leishmaniasis; cases, palmas

Abstract: The infectious diseases are an important problem of the public health, because of the fast propagation by endemics in urban environment associated to migratory pathway country-town, increase of the population density, among other factors. Therefore, is urgent intensify the studies of the human ecology to identify the factor risks, social-economic and from ambient, that cause this disease. The Visceral Leishmaniasis [VL] is a infectious, emergent, endemic and neglected disease in around 65 countries with approximately 500.000 new cases/year, is more incident in India, Bangladesh, Nepal, Sudan and Brazil. The hypothesis of the causes of the dissemination are many and the preventive actions are not efficient where is endemic, therefore, the objective this work was to study the VL in Palmas, Tocantins state in Brazil. According Sinan/SVS/MS data, the Tocantins has the most number of cases of VL in North region and in Brazil, with average of the 418 last 5 years with more incidence in 2011 with 487 confirmed cases, 22 obits, 45% of lethality ratio and 34.8 incidence ratio [the national ratio this year was 2.0]. The data this work indicates that Palmas had presented last 5 years 212 cases of VL, being more prevalent in male [65%], among children from minus 1 to 4 years old [86 cases]. Was observed higher prevalence in 5 of the 65 neighborhood with more incidence in the March, May, January and June months with 22, 21, 20 e 19 cases, respectively. The data from this work indicate that Palmas has 10,1% of the all cases in Tocantins state last 5 years, presenting specific regional factors like high prevalence in alternated months, differently of the others regions where the high prevalence correspond to rainy period. In fact, this was verified this work, but the incidence in children was statistically higher than old peoples. These data indicate that is necessary a specific strategy from health programs in Palmas, focusing the neighborhoods with highest prevalence of the VL and on the children with age inferior to 4 years.

- EPIDEMIOLOGY [945] P 829 - OCCURRENCE OF IGG ANTI-LEISHMANIA SPP ANTIBODIES IN DOGS FROM LOCAL RECYCLING CENTERS AND NEIGHBORHOOD WITH ADJACENT FOREST IN THE URBAN AREA OF LONDRINA, PR, BRAZIL. DA COSTA, L.; RUFFOLO, B.B.; TOLEDO, R.S.; EWALD, M.P.C.; DIAS, R.C.F.; VIDOTTO, O.; MITSUKA-BREGANÓ, R.; NAVARRO, I.T.; FREIRE, R.L. DEPARTAMENTO DE MEDICINA VETERINÁRIA PREVENTIVA, UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL.

Keyword:solid waste; adjacent forest; reservoir

Abstract: The American Cutaneous Leishmaniasis (ACL) is a zoonosis caused by a protozoa of the genus Leishmania, an intracellular parasite of mononuclear phagocyte system cells, spleen, liver, bone marrow, lymph nodes, mucosal and leukocytes. The transmission occurs through the bite of infected sand flies of the genus Lutzomyia. ACL is considered by the World Health Organization one of the six most important infectious diseases in the world. It occurs in all states of Brazil, and in Paraná State is present in the northern and northwestern region and in the Ribeira river valley. Human actions on the environment as the increasing and disorderly urbanization, social and economic changes have helped in expanding endemic areas and outbreaks of ACL in urban

areas. The identification of reservoir hosts of Leishmania spp, like dogs, are important to determine the natural cycle of the parasite and to better understand the epidemiology of the disease. The aim of this work was to study the epidemiology of ACL in dogs residents in recycling of solid waste centers (Group 1) and in dogs from a neighborhood adjacent to a forest (Group 2), in Londrina, Paraná State. Blood samples of 61 dogs from Group 1, and 74 dogs from Group 2 were randomly collected. The research of IgG anti-Leishmania antibodies was performed by Indirect Immunofluorescent Antibody Test (IFAT), using promastigotes forms of L. amazonensis, with a cutoff point of 40. Of the 135 dogs analyzed, 13 animals (9.6%) were positive, nine (14.75%) in Group 1 and four (5.40%) in Group 2. When comparing Groups 1 and 2, there was no statistically significant difference. The recycling of solid waste is a major source of income to the poor, held in a disorganized and informal manner. The results lead us to the conclusion that these areas have antibodies against Leishmania spp circulating in the environment from the five regions of Londrina and in one neighborhood adjacent to a forest.

- EPIDEMIOLOGY [955] P 830 - VISCERAL LEISHMANIASIS-HIV COINFECTION ASPECTS IN NORTHEASTERN BRAZIL. BARBOSA, I.R.; TORRES, Ú.P.S.; TAVARES, A.M. CENTRO DE CONTROLE DE ZOONOSES, NATAL, RN, BRAZIL.

Keyword:visceral leishmaniais; hiv; coinfection

Abstract: Introduction: The increased incidence of visceral leishmaniasis in areas previously unaffected and reemergence in endemic areas are associated with modifications of the environment, migration, disorderly process of urbanization, poverty and mainly to individual risk factors such as expansion of AIDS epidemic. Recent changes in the epidemiological profile of AIDS and visceral leishmaniasis in Brazil point to increased population exposure to both infections, characterized by the internalization of HIV infection concurrent urbanization of visceral leishmaniasis. Despite the need for understanding the magnitude of visceral leishmaniasis-HIV coinfection, there are few studies that address the epidemiological aspects in Northeastern Brazil. The clinical and epidemiologic characterization of this epidemic subsidizes public health policies for the development of prevention strategies, as well as laying the foundations for the medical care provided to patients infected. Objective: To describe the epidemiological profile of cases of visceral leishmaniasis co-infected with HIV (HIV-VL) in northeast Brazil in the years 2007 to 2011. Methods: A descriptive study of cases of visceral leishmaniasis co-infected with HIV live in the Northeast of Brazil, registered in the Information System for Notifiable Diseases (Sinan). Results: Out of 9615 cases of VL, 5.08% were coinfected with HIV, the year with the highest percentage of coinfection was the year 2011 with 7.12% of coinfection, the state of Paraíba had the highest percentage of coinfection, followed Rio Grande do Norte and Alagoas, although in absolute numbers of cases of coinfection, the state of Ceará has obtained the highest number of notifications (n = 175 cases). Males accounted for 81.3% of these cases, the browns (84 2%) and aged 20 to 39 years (56%) were the most affected; 41.5% had complete or incomplete primary education; the lethality rate was 10.6%, and the cure rate was 62% .

Parasitological diagnosis was performed in 69.9% of cases. In 8.49% of the municipalities in the Northeast reported cases of coinfection. Conclusion: These findings emphasize the magnitude of HIV-coinfection visceral leishmaniasis in northeastern Brazil, for although the profile of patients HIV-visceral leishmaniasis coinfected is not different from those with classic visceral leishmaniasis, the greater lethality points to the need for structuring services patient care and health surveillance.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK

[958] P 831 - LEISHMANIASIS VECTORS IN PORTO VELHO-RO: SPECIES MONITORING IN THE INFLUENCE AREA OF SANTO ANTONIO HYDROPOWER PLANT RIBEIRO, K.A.N.1; SENA, J.M.1; GALARDO, A.K.R.2; GRAZINOLLI, M.3 1.SANTO ANTONIO ENERGIA, PORTO VELHO, RO, BRAZIL; 2.INSTITUTO DE PESQUISAS CIENTÍFICAS E TECNOLÓGICAS DO ESTADO DO AMAPÁ -, MACAPÁ, RO, BRAZIL; 3.PROBIOTA CONSULTORIA AMBIENTAL, SAO PAULO, SP, BRAZIL.

Keyword:leishmaniasis vectors; environment; hydropower plant

Abstract: This study is part of the Public Health Monitoring Program from Santo Antonio Hydropower Plant, as condition of Brazilian licensing process. The collects were made between September 2011 and January 2012. Sampling was designed for pre and pos installation periods, in the direct influence area, and sampled 14 points distributed in 3 health regions in Porto Velho. Sampling were carried out during rainy season (January and February), dry season (May and June) and transition season (September and October) during 2011 and 2012. CDC miniature light traps, were placed at three different levels at ground level (soil level, 1.5 m and tree canopy). Between 18:00 PM and 6:00 AM of the following day, at two consecutive nights, in a total of 24 collect hours for each 42 traps . Collections were made from sand flies in indoor and peridomestic environments, with the Castro capturing and manual collections on the walls of homes, during the night species diversity, abundance and frequency were identified for six stages, 2,340 sand flies were collected ( 432 in the first, 243 in the second, 187 in the third, 425 in the fourth, 842 in the fifth and 211 in the sixth). 59 species were collected in the genera Lutzomyia (n=42), Psychodopygus (n=14) e Brumptomyia (n=01). P. davisi was the most abundant species, representing 21% of captured individuals. L. flaviscutellata, vector of Leishmania guyanenses deserves to represented 8% of sand flies captured and L. umbratilis were only 69 individuals (3%). This is the main transmisor specie of LTA, in Amazon region. It was found that was in the abundance of sand flies 182 individuals in the transition period, comparing 2011 and 2012. In 2011, Porto Velho presented 58 american tegumentar leishmaniose cases and 102 in 2012, however the occurrence of cases is not related to the activities of the construction of Santo Antonio Hydropowerplant. There are no autochtonous cases of visceral leishmaniasis, also had no record of presence of its vector L. longipalpis in the region so far.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [963] P 832 - NYCTHEMERAL RHYTHM OF PHLEBOTOMINES (DIPTERA: PSYCHODIDAE) IN QUILOMBOLA AREA, PIRAPUTANGA, STATE OF MATO GROSSO DO SUL, BRAZIL, PRELIMINARY RESULTS INFRAN, J.O.M.1; DE FIGUEIREDO, H.R.1; RIBEIRO, L.M.1; CASARIL, A.E.1; IGUCHI, G.U.1; SOUZA, D.A.1; DE OLIVEIRA, W.A.1; DE OLIVEIRA, E.F.2; OSHIRO, E.T.1; OLIVEIRA, A.G.1 1.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL; 2.FACULDADE DE SAÚDE PÚBLICA DA UNIVERSIDADE DE SÃO PAULO, SAO PAULO, SP, BRAZIL.

Keyword:nycthemeral rhythm ; piraputanga; phlebotomine

Abstract: In Brazil leishmaniases are occurring in all regions of the country. In Mato Grosso do Sul (MS) have been expanding in all municipalities and both American cutaneous leishmaniasis (LT) and visceral leishmaniasis (VL) have a high incidence in Aquidauana. This study is being conducted at the Furna dos Baianos, quilombola area, the District of Piraputanga, Aquidauana municipality. It is located to the left side of Aquidauana River in the mountain range of Maracaju. With about 3 000 hectares, has economic activities of tourism and production of homemade sweets. This study aims at presenting distribution, diversity, abundance, hourly rhythm and anthropophily of species. From June to December 2012, monthly, for 24 uninterrupted captures were performed in Shannon`s trap at the foot of the mountain. Abiotic

factors such as temperature and relative humidity were recorded during this period. The sand flies captured were sent to the Laboratory of Human Parasitology at the Federal University of Mato Grosso do Sul in order to identify and analyze molecular species of Leishmania in the females. A total of 381 sand flies were captured and the following species have been identified so far: Nyssomyia whitmani, Psathyromyia misionensis, Psathyromyia shannoni, Evandromyia evandroi, Evandromyia lenti, Lutzomyia dispar, Lutzomyia longipalpis, Micropygomyia oswaldoi and Micropygomyia termitophila. The most abundant species was Nyssomyia whitmani (261) totaling 67.44%, with 50.19% of females and 49.81% males. The second species was Pa. misionensis (69) with 17.82%, 97.10% of females followed by Pa. shannoni (21) with 5.42% of the total species. The frequency of these insects showed variation over the months. In July, the hours of 5:00am showed greater abundance. However, between August and November most insects was caught between the 17:00pm to 21:00pm, with temperature and humidity ranging between 17°C to 27°C and 63% to 95%, respectively. This study will be conducted for 24 months.

- EPIDEMIOLOGY [968] P 833 - VISCERAL LEISHMANIASIS- TRANSMISSION INTENSE IN MUNICIPAL HIGH RISE AGRIBUSINESS BAHIANO, 2007 TO 2011.

LEITE, M.D.X.1; NASCIMENTO-SILVA, M.M.1; BARREIROS, D.C.2; FRAGA, D.B.M.1; BAVIA, M.E.1 1.UFBA, SALVADOR, BA, BRAZIL; 2.SECRETARIA MUNICIPAL DE SAÚDE DE BARREIRAS, BARREIRAS, BA, BRAZIL.

Keyword:visceral leishmaniasis ; positive index ; control measures

Abstract: The Visceral Leishmaniasis (VL) is classified by the World Health Organization(WHO) as one of the seven major endemic diseases of global impact. In Brazil, the great number of VL cases are aggregated in the northeast of Brazil. Several endemic areas are observed in Bahia.

Barreiras, a city located in Bahia, has shown a high number of VL reports in humans and dogs.

This city has the third largest migration flow of Bahia, due to the strong development of agribusiness in the region. Due to the economic importance of the region and because Barreiras is classified as an area with intense transmission, studies should be conducted to clarify the epidemiology of VL in this municipality. In this study we evaluated the positive index (PI) of canine and human in endemic area of VL. The study area has a sub-humid climate with average annual temperature of 31.15 º C. Notifications of dog cases that occurred in the period of 2007 to 2011 were provided by the Center for Zoonotic Control of Barreiras (CCZ). VL diagnosis was performed using Immunofluorescence Assay test (IFI) in 6623 samples of domicile and stray dogs. VL notifications in humans were provided by the Health Department of the Municipality of Barreiras (SSB). PI were estimated by the confirmed VL cases divided by the number of samples tested by IFI. From the total dog samples tested, 12 % were positive. From the positive dogs, 71% were domicile and 29% stray dogs. The PI of dogs in 2007 was 10%, 2008 (38%), 2009 (24%), 2010 (7%) and 2011 (11%). While in humans, PI was 2007 (9%), 2008 (25%), 2009 (23%), 2010 (30%) and 2011 (12%).The rates of PI varied along the years.

Control measures for VL were intensified in 2009, leading to a reduction of the positive dogs in 2010. The number of positive cases in humans decreased in 2011, probably because of the reduction of the positive dogs in the previous year. These findings indicate that Barreiras presented high rates of VL occurrence and control measures seems to have reduced the ocorrence of VL.

- EPIDEMIOLOGY [978] P 834 - EPIDEMIOLOGICAL SITUATION OF AMERICAN CUTANEOUS LEISHMANIASIS IN THE MUNICIPALITY OF MARAÚ - BAHIA, BRAZIL: 2007 TO 2012 DA COSTA, L.B.; CASTRO, J.M.; SANTOS, R.S. DIRETORIA DE VIGILÂNCIA EPIDEMIOLÓGICA DA SECRETARIA DE SAÚDE DO ESTADO DA BAHIA, SALVADOR, BA, BRAZIL.

Keyword:american tegumentar leishmaniasis; epidemiology; control

Abstract: Introduction: Cutaneous Leishmaniasis (ACL) is a disease caused by protozoa of the genus Leishmania, transmitted to humans by the bite of sand flies (Order Diptera: Family Psychodidae: Sub-Family Phlebotominae). In Brazil there are currently six species of Leishmania responsible for human disease, and more than 200 species of sandflies involved in the transmission. It is a disease that has accompanied man since ancient times and has presented in the past 20 years, an increasing number of cases and expanding its geographic occurrence, being found now in all Brazilian states under different epidemiological profiles, with new focus and outbreaks. It is estimated that the major ocurrence is in the North and Northeast. In Bahia cases reach 245 municipalities, mainly in the area of Costa do Dendê. Objectives: To analyze the evolution of cases of American cutaneous leishmaniasis in the municipality of Maraú, in the period from 2007 to 2012. Methods: The cases were obtained from the Information System for Notifiable Diseases (SINAN) through the application of TABWIN. We analyzed the temporal distribution of the incidence of LTA according to the selected variables: age, sex, clinical presentation and outcome. We did not include cases disposed in the system. Results: 389 cases were reported with more frequency in 2011 (31,1%) and variation of 63,5% compared to 2010, above the average of cases in the period (57.5). Of the 13 health services in the city notifying in the period studied, including emergency care units and family health services, stood out the localities of Ibiaçu and Tremembé, representing 73.8% of cases. Most cases occurred in men (62.2%) and in the cutaneous form (99.7%). Regarding the evolution of the cases we observed that 86.9% (338) were cured and 11.8% did not respond. The incidence in children under 10 years was 15.7% with emphasis on the location of Ibiaçu. The distance between Maraú and municipalities with higher detection rate in Bahia, Ituberá and Taperoá, is 46 km and 65 km, respectively. Conclusion: The increasing rise in cases in the city of Marau, exception in 2010, indicates the need for research in the areas of higher incidence, including the evaluation of epidemiological measures adopted. The occurrence in children alert to the possibility of the presence of the vector indoors and intensification of preventive and control measures, and implementation of health education activities aimed at vulnerable groups.

- OTHER SPECIAL TOPICS [987] P 835 - GENETIC DIVERSITY IN LEISHMANIA (VIANNIA) BRAZILIENSIS FROM AN OUTBREAK OF CUTANEOUS LEISHMANIASIS: POTENTIAL CLINICAL AND EPIDEMIOLOGICAL IMPLICATIONS. DE LOS SANTOS, M.B.1; VALDIVIA, H.V.2; ORE, M.3; SAENZ, E.4; CABRERA, R.5; FERNANDEZ, J.3; SANCHEZ, J.F.1; BALDEVIANO, G.C.1; EDGEL, K.A.1; LESCANO, A.G.1

1.DEPARTMENT OF PARASITOLOGY, U.S. NAVAL MEDICAL RESEARCH UNIT 6, LIMA, PERU; 2.LABORATORIO DE IMUNOLOGIA E GENOMICA DE PARASITOS, UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 3.INTELIGENCIA SANITARIA, COMANDO DE SALUD DEL EJÉRCITO, LIMA, PERU; 4.SERVICIO DE DERMATOLOGÍA, HOSPITAL MILITAR CENTRAL CORONEL LUIS ARIAS SCHREIBER, LIMA, PERU; 5.DIRECCIÓN GENERAL DE EPIDEMIOLOGÍA, MINISTERIO DE SALUD, LIMA, PERU.

Keyword:leishmania (viannia) braziliensis; genetic diversity; epidemiology

Abstract: Outbreaks of leishmaniasis with narrow exposure windows can be good opportunity to understand the diversity of circulating strains in endemic settings. We studied the intrapopulation genetic diversity and genetic clusters of leishmaniasis outbreak that affected Peruvian Army cadets, in the northwestern Peruvian Amazon Basin.

We established 33 L. (V.) braziliensis cultures from promastigotes isolated from outbreak cases using blood agar medium. DNA was extracted and purified from parasite cultures using the DNeasy Blood and Tissue kit (QIAGEN) and we used 10 pairs of labeled primers (AC01, AC52, G09, E11, CAK, Ibh3, LBA, EMI, LRC and ARP) previously reported by Rougeron et al. PCR reactions were performed in an Applied Biosystems GeneAmp PCR System 9700 thermocyclers and PCR products were processed by an Applied Biosystems 3130xl Genetic Analyzer. We used GeneMapper v4.0 software for discrimination of electropherogram peaks.

To determine genetic parameter of variability we used Cervus 3.0 and Hardy-Weinberg deviation was estimated with a score test and Fisher’s method using Markov chain algorithm by Genepop v4.0. Structure v2.2 was used for Bayesian grouping analysis and Stata v12.1 to analyze the correlations between genetic clusters and clinical data.

We identified 52 microsatellite alleles in 33 Leishmania strains (5.2 mean alleles per locus) with mean observed (Ho) and expected (He) heterozygosity of 0.649 and 0.625, respectively, and polymorphic information content (PIC) of 0.573. Additionally, we determined that 9 of the 10 loci did not show significant deviation from the Hardy-Weinberg equilibrium; only the CAK locus was in deviation by heterozygote deficit (p<0.05). Finally, Bayesian grouping analysis showed two groups or genetic clusters (K=2, Pr(X/K)=1.0). The two clusters shown marginallysignificant, differences in the number of lesions per patients (p=0.138), the presence of ulcers (p=0.053), and plaques (p=0.104), despite the small sample size.

Our findings suggest significant parasite genetic diversity with potential clinical implications even after a short time of exposure in a single geographical location, and underline the importance of microsatellites markers to understand leishmaniasis’ epidemiology.

- EPIDEMIOLOGY [988] P 836 - VISCERAL LEISHMANIASIS AT THE SEMIARID REGION OF WESTERN VENEZUELA AÑEZ, N.1; ROJAS, A.1; VARGAS, E.2; MEDINA, V.2; CRISANTE, G.1; YEPEZ, J.Y.2 1.UNIVERSIDAD DE LOS ANDES, MERIDA, VENEZUELA; 2.UNIVERSIDAD FRANCISCO DE MIRANDA, CORO, VENEZUELA.

Keyword:visceral leishmaniasis; semiarid region; venezuela

Abstract: An epidemiological study on visceral leishmaniasis (VL) at the semiarid region of western Venezuela, where some acute cases have been previously reported, is recorded. A total of 1036 symptomless people sera samples from 25 villages, and 67 dogs from 8 different localities, were tested by using 3 different serologic methods and the polymerase chain reaction assay.

Serological tests revealed 16.9% and 76.1% seropositive to L.infantum in asymptomatic humans and dogs, respectively. In addition, a PCR assay detected the presence of L.infantum-specific DNA in 17.7% of the symptomless sampled individuals and in 29.9% of the tested dogs. The combined analyses of serologic and molecular findings demonstrate the presence of subclinical or inapparent VL infections in local people from this part of western Venezuela. The present results also serve to call the attention on the presence of infected dogs as a risk factor in the

maintenance of L.infantum as a source for infection to humans in the studied semiarid region. A total of 12 sand fly species were identified from 6000 specimens collected in 12 sampled villages, with a predominance of Lu.evansi (66.6%) over Lu.longipalpis (14.9%). In both sand fly species natural infection by L.infantum was detected using a PCR assay. It is concluded that the association among VL infected people, L.infantum-infected dogs and the presence of Lu.evansi, together with the observed poor sanitary and/or nutritional conditions, are risk factors to explain the high endemicity by VL at the semiarid region of western Venezuela.

- CLINICAL LEISHMANIASIS [989] P 837 - FELINE LEISHMANIOSIS IN ITALY: RETROSPECTIVE EVALUATION OF 24 CLINICAL CASES PENNISI, M.G.1; LUPO, T.2; MIGLIAZZO, A.2; PERSICHETTI, M.F.1; MASUCCI, M.1; VITALE, F.2 1.DIPARTIMENTO DI SCIENZE VETERINARIE, UNIVERSITĂ&#x20AC; DI MESSINA, ITALIA, MESSINA, ITALY; 2.CENTRO DI REFERENZA NAZIONALE PER LA LEISHMANIOSI (CRENAL), IZS-SICILIA, PALERMO, ITALY.

Keyword:cat ; feline leishmaniosis; leishmania infantum

Abstract: Clinical leishmaniosis is sporadically reported in cats living in endemic areas and the feline disease is still not well known. The aim of this study is to give information on the clinical features of the natural disease caused by Leishmania infantum in cats. Data were obtained from 24 clinical cases in which the diagnosis was confirmed by at least one of the following methods: IFAT, PCR, isolation. In some cases cytological or histological evaluations were also available.

The disease was diagnosed in 23 domestic shorthaired and 1 Persian cats, 11 females and 13 males, age range 2-14 years (mean 9, median 10). Sixteen cats lived fulltime or partly outdoor, 4 lived indoor (3 in a rescue cattery and 1 in apartment), 2 others were stray cats. The reason for consultation was weight loss (8 cases), anorexia (7), ulcerative/nodular dermatitis (6), chronic upper respiratory tract disease (4), chronic gingivo-stomatitis (4), apathy (3), diarrhea (3), lymph node enlargement (3), itching (3), spaying (2), lethargy (1), polyuria/polydipsia (1), polyphagia (1), dyspnea (1), uveitis (1), conjunctivitis, (1), otitis (1) trauma (1). Physical examination main findings were: lymph node enlargement (18 cases), ulcerative dermatitis (7), emaciation (6), chronic gingivo-stomatitis (6), dehydration (6), conjunctivitis (4), uveitis (3), nodular dermatitis (2), pale mucous membranes (2), tracheitis (2), otitis (3), hemorrhagic nodule (1), crusty dermatitis (1), pleural effusion (1), liver enlargement (1), jaundice (1), spleen enlargement (1), thyroidal nodule (1), chorioretinitis (1), alopecia (1). All cats had antibodies reacting with L. infantum and IFAT titer ranged between 1:10 and 1:5120. PCR was performed on blood, lymph node or other tissues in 22 cats and 21 tested positive on at least one tissue. Isolation was obtained from 7 cats. Cell blood count abnormalities were found in 15 cats out of 22 and anemia was the most common finding (55%). Twenty-three cats were serologically tested for Feline Immunodeficiency Virus (FIV) and Feline Leukemia Virus (FeLV) infections: 12 were negative for both agents, 10 were positive for FIV and 1 for both FIV and FeLV infections. According to biochemical investigations performed in 20 cats, chronic renal failure was diagnosed in 4 individuals, diabetes mellitus in one and hepatic lipidosis in one other cat. Feline leishmaniosis is mainly diagnosed in old, outdoor cats presenting lymph node enlargement and frequently coinfected by FIV. - VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [991] P 838 - APLLICATION OF MODIFIED CDC AND DISNEY TRAPS FOR CAPTURE OF SANDFLIES IN AN AREA OF FISHING TOURISM, BONITO MUNICIPALITY, MATO GROSSO DO SUL, BRAZIL NUNES, V.L.B.1; BRILHANTE, A.F.1; CRISTALDO, G.2; DORVAL, M.E.M.C.2; RIBEIRO, P.F.M.3

1.UNIVERSIDADE ANHANGUERA- UNIDERP, CAMPO GRANDE, MS, BRAZIL; 2.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL; 3.UNIVERSIDADE ANHANGUERA-UNIDERP, CAMPO GRANDE, MS, BRAZIL.

Keyword:phlebotomines; evandromyia sp.; traps

Abstract: The purpose of this study was to investigate the sandfly fauna at a gallery forest with evidence of secondary vegetation following the selective extraction of timber in a fishing tourism area located in the municipality of Bonito, Mato Grosso do Sul, Brazil, using two kinds of traps. The captures were performed monthly using modified automatic light CDC, from 6 p.m. to 6 a.m., from December 2009 to November 2010, and with a modified Disney trap installed 20 centimeters from the ground, using as bait a rodent (Mesocricetus auratus), from November 2010 to October 2012. All the female specimens captured were dissected for investigation of flagellated forms and identification of the phlebotomine species. The male sandflies were kept over refrigeration until their clarification. The identification was undertaken in according with

by Galati (2003). During the study period of the CDC trap were captured 66 specimens, 24 males and 42 females: Brumptomyia avellari (1), Br. brumpti (2), Evandromyia sp.

(Cortelezzii complex) 24), Ev. lenti (3), Lutzomyia longipalpis (5), Nyssomyia whitmani (9), Psathromyia hermalenti (1), Pa. punctigeniculata (1), Pa. shannoni (19) and Sciopemyia sordelli (1). With the Disney trap 718 insects were captures, 253 males and 465 females: Evandromyia sp. (715), Micropygomyia quinquefer (1), Ny. neivai (1) and Ny. whitmani (1). The greatest phlebotomine species richness was observed with the CDC captures although Disney’s were more expressive, mainly for the Cortelezzii complex specimens who were uncovered as rodentophilic habits. All three specimens that compose this complex are recurrent in all states of the Midwest Region, and Ev. corumbaensis being the only one restricted to this region. Due to epidemiological value recently assigned to the Ev. cortelezzii and Ev. sallesi, found naturally infected with Leishmania in Minas Gerais, the current allowance of sandflies as regards to infection through Leishmania sp. as well as the fact that the studied area has fishing tourism as the major business and recreational activity, behavioral and ecological studies of these insects are required with the aim to promote actions of inspection and prevention of cases of cutaneous leishmaniasis.

- RESERVOIRS [993] P 839 - RELATIONSHIP BETWEEN CLINICOPATHOLOGICAL FINDINGS AND PARASITE BURDEN IN DOGS WITH VISCERAL LEISHMANIASIS FROM AN ENDEMIC AREA OF BRAZIL DA SILVA, S.M.1; AMORIM, I.F.2; RIBEIRO, R.R.3; DE PAULA, R.C.4; AZEVEDO, E.G.2; DEMICHELI, C.5; MELO, M.N.2; TAFURI, W.L.2; MICHALICK, M.S.2; FREZARD, F.5 1.INSTITUTO DE CIÊNCIAS BIOMÉDICAS, FEDERAL UNIVERSITY OF UBERLÂNDIA, UBERLÂNDIA, MG, BRAZIL; 2.INSTITUTO DE CIÊNCIAS, , FEDERAL UNIVERSITY OF UBERLÂNDIA BIOLÓGICAS, BELO HORIZONTE, MG, BRAZIL; 3.CENTRO DE CIÊNCIAS AGRÁRIAS, AMBIENTAIS E BIOLÓGICAS, FEDERAL UNIVERSITY OF RECONCAVO DA BAHIA, CRUZ DAS ALMAS, BA, BRAZIL; 4.FACULDADE DE FARMÁCIA, FEDERAL UNIVERSITY OF MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 5.INSTITUTO DE CIÊNCIAS BIOLÓGICAS, FEDERAL UNIVERSITY OF MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL.

Keyword:canine visceral leishmaniasis; clinical stage; parasite burden

Abstract: Visceral leishmaniasis (VL) is a neglected disease, closely associated to poverty which causes thousands of deaths among human beings worldwide every year. In Brazil VL is a zoonosis caused by Leishmania infantum and domestic dog are the main urban reservoir of the parasite.

The disease in dogs is characterized by a marked pleomorphism, and the clinical signs vary according to the immune response of the animals toward the infection. Other conditions such as genetic susceptibility, nutritional status, immunosuppression, co infections, co morbidities,

strain virulence and parasitic burden can contribute to the severity of the disease. In general, the main clinical signs of canine visceral leishmaniasis (CVL) are various degrees of dermopathy, lymphadenopathy, onychogryposis, weight loss, abnormalities of the musculoskeletal system, eye lesions, hematopoietic disorders, renal disease, and lesions originating from immune complex deposition in tissues. This work aimed to evaluate the clinicopathological findings and its association with parasitic burden in dogs with CVL from the Metropolitan Region of Belo Horizonte (Minas Gerais State, Brazil), one of the most endemic area for VL and CVL in Brazil.

Fifty two dogs with positive results in ELISA and IFAT (indirect immunofluorescence assay) were submitted to physical evaluation followed by laboratorial tests: complete blood count, serum biochemistry, immunohistochemistry (IHQ) and qPCR in ear skin fragment, bone marrow and ultra sound guided spleen aspirates. Based on serological, physical and clinicopathological findings, dogs were classified in clinical stages of CVL (I-mild, II-moderate, III-severe, IV-very severe). Spearman´s correlation test (CI=95%) was used to evaluate the association between the variables. Parasite burden in skin, bone marrow and spleen were positively associated with each other and with anti-Leishmania IgG titers, total serum proteins and globulins, and negatively associated with red blood count, hematocrit and platelets. The parasite burden in bone marrow also was positively associated with monocytes count, and skin and spleen burdens were negatively associated with eosinophils count. The presence of parasites in the skin determined by IHQ was positively associated with skin parasitic burden (qPCR) and clinical staging. Clinical staging showed 92.3% of the animals in stages II or III and 7.7% in stage I (mild disease). The clinical stage was positively associated with the physical examination, anti-Leishmania IgG titers, presence and burden of parasites in skin and spleen.

Based on these findings, in our opinion, clinicians should adopt the staging of CVL in routine as an efficient tool to establishment of accurate prognosis, treatment protocol and follow-ups.

Financial support: CNPq, FAPEMIG and CAPES. - EPIDEMIOLOGY [996] P 840 - PALEOEPIDEMIOLOGY OF LEISHMANIA GENUS (KINETOPLASTIDA, TRYPANOSOMATIDAE) NOVO, S.P.C.1; LELES, D.2; DUTRA, J.M.1; ARAÚJO, A.1

1.FUNDAÇÃO OSWALDO CRUZ - FIOCRUZ / ESCOLA NACIONAL DE SAÚDE PÚBLICA SÉRGIO AROUCA - ENSP / DEPARTAMEN, RIO DE JANEIRO, RJ, BRAZIL; 2.UNIVERSIDADE FEDERAL FLUMINENSE - UFF / INSTITUTO BIOMÉDICO, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:paleoepidemiology; leishmania sp.; molecular diagnosis

Abstract: Species of the genus Leishmania (Kinetoplastida, Trypanosomatidae) are causative agents of leishmaniasis. The identification of these parasites in archaeological material is performed by molecular diagnosis. Analysis of mitochondrial DNA from Leishmania sp. in samples from 90 mummies of the pre-Egyptian dynasty, the dynasty of Abydos (3500-2800 BC) and from western Empire (2050-1650 BC), identified the compatibility of the archeological DNA sample with modern DNA of Leishmania donovani, an agent of visceral leishmaniasis (Zink et al, 2006). Recently, molecular studies and immunological tests have been made, diagnosing the presence of Leishmania infantum in bone fragments of Eleonora de Toledo (1522-1562) (Nerlich et al, 2012, Bianucci et al, 2012). The present work aims to approach the paleoepidemiology of Leishmania sp. in the ancient cultures. For this, we will use samples of tissues and bones fragments from human and other animals stored in our laboratory, collected from South American archaeological sites. We will submit the samples to the PCR amplification and subsequent sequencing. The identified sequences will be confronted with sequences deposited in GenBank. The aim is to obtain data on the distribution of this infection over time in the regions studied, analysis of the origin and antiquity of the host-parasite relationship and analysis of nucleic acids of the parasite with the purpose of evolutionary studies. To test the

effectiveness of ancient DNA extraction kit, four samples extracted from natural mummified bodies were used. The bodies were found in Minas Gerais state, Brazil, dating from the colonial period (circa 1700 AD). To verify the current inhibitors in the samples , human DNA amplification was performed and to verify Leishmania sp. molecular target to Kinetoplast minicircule kDNA was used. The results showed that there was no inhibition and sequences of two samples show similarity with Leishmania sp.

Financial support: CNPq and FAPERJ.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [998] P 841 - STUDY OF SAND FLY FAUNA COMMUNITY IN AN ENDEMIC RURAL AREA OF AMERICAN CUTANEOUS LEISHMANIASIS IN THE MUNICIPALITY OF MOGI GUAÇUSP. COLLA-JACQUES, F.E.1; CASANOVA, C.2; PRADO, A.P.D.1 1.INSTITUTO DE BIOLOGIA, UNIVERSIDADE ESTADUAL DE CAMPINAS, CAMPINAS, SP, BRAZIL; 2.SUPERINTÊNDENCIA DE CONTROLE DE ENDEMIAS, SECRETARIA DA SAÚDE DO ESTADO DE SÃO PAULO, SÃO PAULO, SP, BRAZIL.

Keyword:sand flies; american cutaneous leishmaniasis; são paulo state

Abstract: From the six sand fly species considered important for the American cutaneous leishmaniasis (ACL) in São Paulo state, two of them, Nyssomyia neivai and Nyssomyia intermedia, have been regularly found in high abundance in areas where the disease has been reported. Mogi-Guaçu river valley, located in the Northeast region of São Paulo state, is endemic for ACL and in most cases, N. neivai has been by far the most predominant species. On the other hand, recent entomological investigation for confirming the autoctony of two ACL cases in a rural area of Mogi Guaçu, has reported the high abundance of Nyssomyia whitmani, Expapillata firmatoi and Migonemyia migonei, and nonappearence of N. neivai. Aiming at evaluating the role of each one of this species in the transmission cycle of the disease, the spatial and monthly distribution of the sandflies was studied, as well as the natural infection by Leishmania braziliensis, in different environment where the cases were registered. To evaluate the role of each environment in maintaining the disease cycle and risk of transmitting it, the sandfly community was characterized by using several ecological indices, such as richness, abundance, diversity and similarity. During the studied period, between October/2010 and April/2012, five CDC automatic light traps were installed weekly from 6 p.m. to 8 a.m. in four different kinds of environment: inside the woods, in its edge, in a chicken shed and nearby the human dwelling.

More than mais de 10,000 sandflies were collected, distributed in 15 species. Out of the total, close by 50% were N. whitmani, followed by Mg. migonei and Ex. firmatoi approximatelywith 30 and20%, respectivamente. The chicken shed was the environment where the greatest number of sandflies were collected (more than 60% of the collected flies). However, about 5% were collected in the human dwelling. From the 1,622 females dissected two, a Mg. migonei and an Ex. firmatoi, collected in the chicken shed were found promastigote forms. The sandfly community caracterization by the indices of richness, abundance, similarity and diversity allows us the confirm that there is a risk of getting the disease in every one of the studied environments, and that N. whitmani, Mg. migonei and E. firmatoi may be taking part in maintain the ACL wild cycle. The presence of human cases in the area, added to the presence of important species to the ACL cycle represents the risk of new cases to the area.

- EPIDEMIOLOGY [1001] P 842 - EPIDEMIOLOGICAL SURVEY OF CANINE VISCERAL LEISHMANIASIS IN PARAIBA STATE HINTERLAND, BRAZIL. MELO, M.A.1; FARIAS ROTONDANO, T.E.2; BARBOSA, W.O.1; PINTO, N.F.S.1; LIMA, E.F.1; SOUZA, H.C.1; SEMIÃO-SANTOS, S.J.3; ANDRADE, P.P.2 1.UNIVERSIDADE FEDERAL DE CAMPINA GRANDE, PATOS, PB, BRAZIL; 2.UNIVERSIDADE FEDERAL DE PERNAMBUCO, RECIFE, PE, BRAZIL; 3.UNIVERSIDADE DE ÉVORA, ÉVORA, PORTUGAL.

Keyword:northeast; kala-azar; tick

Abstract: Canine visceral leishmaniasis is a chronic infectious disease and an important zoonosis in the Paraiba State (Brazil) and Leishmania chagasi is found infecting both man and dog. This study aimed to conduct a seroepidemiological study of canine visceral leishmaniasis in three Paraíba State municipalities: Cajazeiras, Sousa and Uiraúna. Seroprevalence was tested by ELISA S7®, whose antigen is the carboxy-terminal region of L. chagasi Hsp70. A questionnaire was applied to owners for identification of risk factors. The seroprevalence found in Cajazeiras, Sousa and Uiraúna were 19.4%, 10.1% and 15.5%, respectively. The risk factors identified in the multivariate analysis by logistic regression was the presence of ticks (Odds Ratio = 3.6276, 95% CI = 1.71 to 7.71, p = 0.0008). The seroprevalence of canine visceral leishmaniasis is high in Cajazeiras, Sousa and Uiraúna and additional studies are needed to evaluate the role of ticks as risk factor.

- EPIDEMIOLOGY [1005] P 843 - EPIDEMIOLOGICAL OVERVIEW OF VISCERAL LEISHMANIASIS IN ALBANIA IN 2002-2011 BINO, S.; ALLA, L.; VELO, E.; MYRSELI, T. INSTITUTE OF PUBLIC HEALTH, TIRANA, ALBANIA.

Keyword:leishmaniasis; epidemiology; prevalence

Abstract: Introduction: Visceral Leishmaniasis is endemic in Alabania but its occurrence in Albania has shown a downward trend over the recent years Aim: To present an epidemiological overview of Visceral Leishmaniasis in Albania over the period 2002-2010, according to relevant registered variables of infected persons (gender, age, living residence, transmission, occupation).

Material and Methods: Elaboration of data reported to National Institute of Public Health – Albania, from District Epidemiological Services and University Hospital Centre as well as data obtained from epidemiological questionnaires and field investigations.

Result: Visceral Leishmaniasis (VL) is endemic in Albania. 668 human cases have been reported over the period 2002 to 2011. The incidence rate in 2002 and 2011 was respectively 4.2 cases/105 population and 1.0 cases /105 population respectively. VL is present in 80% of the country districts. The incidence during years in Lezha district varied from 2.9 cases/10.000 in 2002 to 0.2 cases/ 10.000 in 2011. Shkoder in the northern part of the country showed 1.4 cases/10.000 and Berat in the south with 1.1 /10.000 cases.

Meanwhile Tirana showed a lower one of 0.4/10.000 cases.Sporadically cases were reported in other districts respectively to Kruja, Puka, Mat, Mirdite, Sarande . Most of the reported cases, (71,5%), belong to the age-group 0-14 years old. The most infected were males with 64 % percent of reported cases. Only 70% of reported cases were laboratory confirmed. The diagnosis for other cases was based on clinical signs and symptoms due to lack of laboratory capacities. Rural areas were most affected with 67.3% of the total cases reported.

Conclusions: Leishmaniasis continues to be endemic in Albania with a geographical spread and a high prevalence in the north and central areas of the country. The timely clinical diagnosis is crucial for the initiation of medical treatment of the patients.

- EPIDEMIOLOGY [1009] P 844 - SENTINEL CHICKEN SEROCONVERSION AS A BIOMARKER OF EXPOSURE TO SAND FLY BITES AT AN ENDEMIC AREA FOR VL SOARES, B.R.1; SOUZA, A.P.A.1; PRATES, D.B.2; OLIVEIRA, C.I.1; VALENZUELA, J.3; BARRAL-NETTO, M.1; MIRANDA, J.C.1; BARRAL, A.1 1.CPQGM-FIOCRUZ, SALVADOR, BA, BRAZIL; 2.DEPTO. DE BIOMORFOLOGIA -ICS - UFBA, SALVADOR, BA, BRAZIL; 3.NIAID - NIH, ROCKVILLE, UNITED STATES.

Keyword:lutzomyia longipalpis; igy ; chicken

Abstract: Visceral Leishmaniasis (VL) is transmitted by Lutzomyia longipalpis, during the blood meal, when saliva and parasites are injected into host skin. Chickens in VL endemic area play an important blood food source and their presence is often cited as a risk factor to VL transmission.

However, the role of chickens in the disease epidemiology has not been well defined. We tested here if chicken antibodies against salivary gland sonicate (SGS) could be used as exposure markers to VL and also investigated the potential role of chickens as sentinels. Chickens were distributed in different houses in Cavunge, a VL endemic area in Bahia, and kept to natural exposition to sand flies and monitoring of the seroconversion to SGS was evaluated by ELISA. Experimental immunization with Lu. longipalpis SGS lead to the production of anti-SGS IgY antibodies in all immunized chicken and after the second immunization there was a statistically significant increase in antibody levels. Similarly, chicken naturally exposed to Lu. longipalpis sand fly bites at the VL endemic area also developed significant anti-SGS IgY antibodies.

Beside SGS we observed that rLJM17 and rLJM11 salivary recombinant proteins showed similar performance as SGS. Taken together, these results may be used as important indicators of the Lu. longipalpis vector presence in VL endemics areas, raising the possibility of using the chicken as a sentinel animal.

- EPIDEMIOLOGY [1011] P 845 - AMERICAN CUTANEOUS LEISHMANIASIS AND OCCUPATIONAL HEALTH: AN ANALYSIS OF THE RELATIONSHIP BETWEEN OCCUPATION AND THE INCIDENCE OF CASES IN A CAPITAL OF THE AMAZON REGION. AZEVEDO, F.S.1; GADELHA, S.M.S.2; FALCHETTI, F.1; MONTEIRO, J.C.1 1.FEDERAL UNIVERSITY OF RONDONIA, PORTO VELHO, RO, BRAZIL; 2.APARICIO CARVALHO INTEGRATED COLLEGE, PORTO VELHO, RO, BRAZIL.

Keyword:american cutaneous leishmaniasis; occupational health; agricultural worker

Abstract: This study, a descriptive retrospective quantitative one, analyzes the relationship between American Cutaneous Leishmaniasis (ACL) and the user occupation, in the city of Porto Velho, in Rondonia. One of the risk factors of ACL is the contact with forest areas, pointing out the agricultural workers as the population at risk for ACL, since this is the natural environment and a locus of work. The objective of this work is to identify cases of ACL in which the occupation user may be a predisposing factor to injury, contributing to the prevention of these economic sectors. This study used as the source the database of the Municipal Health Department of Porto Velho/RO, with cases reported from 2007 to 2011. In this period, 455 cases of ACL were reported, being 93.2% (424) new cases. Of the 455 cases, 87.7% are male, and 64.17% are

between 20 and 49 years. Among the 455 cases reported, only in 274 was appointed the occupation of the user. Among these, the occupation farmer/agricultural worker was the most pointed (29.2%). About the relationship of ACL with work, 41.9% users, of the 455 cases registered, contracted the disease at work, and in the year of 2008 there was the higher number of reported cases related to work. The remaining cases, 28.2% said they have no relationship to work, possibly being exposed to cases related to forests or household transmission, and in 29.9% of cases have been not recorded this relationship. It can conclude, after this study, that the ACL is still shaping up as a disease with high incidence rate in Porto Velho/RO, affecting individuals of working age and has a strong relationship with the work done by the patient, which indicates the needing of surveillance and monitoring of populations that develop agricultural work. For this, it is important to valorize filling the field of user’s occupation in reporting a case of ACL, because it is through this information that may be the relationship between illness and work, which will guide the prevention and control of cases from the population of agricultural workers.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1015] P 846 - ECOLOGY OF PHLEBOTOMINAE (DIPTERA: PSYCHODIDAE) IN AQUIDAUANA, ENDEMIC AREA FOR LEISHMANIASES, MATO GROSSO DO SUL, BRAZIL. PRELIMINARY RESULTS FIGUEIREDO, H.R.1; RIBEIRO, L.M.2; INFRAN, J.M.3; CASARIL, A.E.1; IGUCHI, G.U.3; SILVA, D.N.4; MENDES, A.R.4; QUARESMA, P.F.5; ANDRADE FILHO, J.D.5; OLIVEIRA, E.F.6; OLIVEIRA, A.G.1; OSHIRO, E.T.

1.PROGRAMA DE PÓS-GRADUAÇÃO EM DOENÇAS INFECCIOSAS E PARASITÁRIAS, UFMS, CAMPO GRANDE, MS, BRAZIL; 2.PROGRAMA DE PÓS-GRADUAÇÃO EM BIOLOGIA ANIMAL, CAMPO GRANDE, MS, BRAZIL; 3.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL; 4.CENTRO DE CONTROLE DE VETORES, AQUIDAUANA, MS, BRAZIL; 5.CENTRO DE PESQUISAS RENÉ RACHOU, FIOCRUZ, BELO HORIZONTE, MG, BRAZIL; 6.FACULDADE DE PÓSGRADUAÇÃO EM SAÚDE PÚBLICA, USP, SÃO PAULO, SP, BRAZIL.

Keyword:aquidauana; sand flies; ecological aspects

Abstract: Leishmaniases are considered diseases of high importance in public health and are expanding throughout the Brazilian territory. The State of Mato Grosso do Sul registered in 2011, 273 cases of visceral leishmaniasis (VL) and 112 cutaneous Leishmaniasis (CL). The municipality of Aquidauana reported 16 cases and 2 deaths from these diseases. This study aims to identify aspects of the behavior of sand flies (Diptera: Psychodidae: Phlebotominae) in the Municipality of Aquidauana, endemic area for leishmaniases. The captures were carried out in six urban sites, quinzenal, with light automatic traps at 06:00pm to 07:00am and manual aspiration, in peridomicile and intradomicile environments in each dwelling, from April to November 2012.

The specimens were transported to the Laboratory of Parasitology of the Federal University of Mato Grosso do Sul (UFMS) for assemblage, identification, and other observations. After that, females were analyzed by molecular technique for Leishmania sp. The 78 captures yielded 1,254 specimens of phlebotomines, 1092 males (87.08%) and 162 females (12.91%). The fauna was composed of seven species: Evandromyia aldafalcaoae, Evandromyia cortelezzii, Evandromyia evandroi, Evandromyia lenti, Evandromyia walkeri, Lutzomyia longipalpis, Psathyromyia shannoni. The most abundant species was Lu. longipalpis representing 94.09% (1,180), 88.98% males and 11.05% females. The second species was Ev. walkeri with 2.31% (29), 55.17% males and 44.82% females. Overall, the sand flies were more frequently captured in peridomicile 94.33% (1,183) than in the intradomicile 5.42% (68) of the residences. There was predominance of insects in Pinheiro neighborhood, totaling 76.07%. The captures performed with manual aspiration have presented four times more insects than light traps. At the moment all females were negative for Leishmania sp. This study is being conducted for 24 months.

- EPIDEMIOLOGY [1019] P 847 - A COMPARATIVE STUDY OF THE INCIDENCE OF AMERICAN CUTANEOUS LEISHMANIASIS IN THE CITY OF PORTO VELHO, THE STATE OF RONDONIA AND BRAZIL, FROM 2007 TO 2011.

AZEVEDO, F.S.1; BAIÃO, A.1; FERNANDES, L.S.1; GADELHA, S.M.S.2; OLIVEIRA, A.P.M.1; SILVA JÚNIOR, C.F.1 1.FEDERAL UNIVERSITY OF RONDONIA, PORTO VELHO, RO, BRAZIL; 2.APARICIO CARVALHO INTEGRATED COLLEGE, PORTO VELHO, RO, BRAZIL.

Keyword:american cutaneous leishmaniasis; amazon region; epidemiology

Abstract: The American Cutaneous Leishmaniasis (ACL) is an infectious, non-contagious, chronic evolution disease, caused by protozoa of the genus Leishmania, of vector transmission. It is considered an autochthonous disease of the Americas, predominantly in regions of hot and humid weather. During the period from 1991 to 2010, showed an annual average of 27,374 reported cases and an average detection rate of 1.6 / 10,000, constituting a public health problem. The northern region has contributed to the highest number of cases (about 36.4% of total cases registered in the period from 1991 to 2010) and with higher average coefficients of 8/10.000 inhabitants, related to the recent process of colonization. The ACL, considered of universal susceptibility, behaves as an occupational disease associated with economic activities such as mining, agriculture and extraction. The objective of this study is to analyze and compare the incidence of ACL in Brazil and in the Amazon region, especially in the State of Rondonia and the city of Porto Velho, in the years from 2007 to 2011. This study is a retrospective descriptive quantitative study. The survey was conducted through the database of the Municipal Health Department of Porto Velho/RO, and data collection relating to cases of Rondonia and Brazil, in Database Health System. From 2007 to 2011 were 90,211 reported cases of ACL in Brazil, with an average of 18,042.2 cases / year and standard deviation of 664.0. There were 85,440 new cases with an incidence of 4.4 / 10,000. In the same period, were reported 4780 cases of ACL in Rondonia state, with an average of 956 cases / year and standard deviation of 143.4. There were 4526 new cases and incidence of 28.7 / 10,000. In the city of Porto Velho, were reported in this period 455 cases, with an average of 91 cases / year and a standard deviation of 22.3. The new cases totaled 424 resulting in an incidence of 9.7 / 10,000. Facing the data exposed, it can analyze that the incidence of ACL per 10,000 inhabitants in Porto Velho and Rondonia are proportionally higher than the Brazilian average, as was expected. Rondonia had a greater number of cases compared to 10,000 inhabitants than Porto Velho because of the endemic area and the population are much larger than the capital, showing that the interior cities contribute greatly to the high incidence of ACL cases throughout the state of Rondonia.

- EPIDEMIOLOGY [1025] P 848 - ANALYSIS CLINICAL-EPIDEMIOLOGICAL OF CASES OF VISCERAL LEISHMANIASIS IN PREGNANT WITH HIV, BRAZIL GOES, J.J.; CORTEZ, A.M.C.; ALEXANDRA, W.A.; PEDROSA, C.M.S.; CORTEZ, L.R.C.; FREITAS, I.S.; PEIXOTO, V.S. UFAL, MACEIÓ, AL, BRAZIL; .

Keyword:visceral leishmaniasis; hiv coinfection; pregnant

Abstract: Visceral Leishmaniasis (VL)-HIV coinfection becomes increasingly common, especially for ruralization of AIDS and urbanization of the VL, thus becoming an emerging problem. In pregnant, this problem becomes even more pronounced due to physiological

immunosuppression caused by the pregnancy. Visceral leishmaniasis in pregnancy is considered rare, with few publications, especially the LV-HIV coinfection.. During the period from 2007 to 2011, were reported 367 cases of LV in pregnant women in Brazil , reported in the Information System for Notifiable Diseases (SINAN) , Ministry of Health of Brazil, with 28 cases of coinfection VL / HIV. Of the 28 cases examined, three were recurrences. About schooling was registered a patient with higher education, one with high school, and 21 with unknown education. The predominant age group was the reproductive age, with 13 cases. Looking at the geographical region, the Southeast reported 15 cases. The identification of Leishmania was possible in 15 cases, the others were used immune criteria and clinical-epidemiological. There was evolution to death by VL in five cases. Observed the presence of co-infection VL / HIV in 7.6% of reported cases, with a death rate of 17.8%, higher than that found in the general population (5-10%). The importance of co-infection in pregnant women occurs by the physiological changes during pregnancy, predisposing increased risk of infections, and the possibility of vertical transmission. It is necessary to health professionals to be alert of the existence of irregular fever in HIV-positive women of childbearing age, especially in endemic areas of VL, especially during the prenatal period, because of the severity of these diseases when associated. There is a real need to expand the studies on this topic and the lack of studies in endemic regions supposes underdiagnosis or no notification.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1030] P 849 - NATURAL BREEDING SITES OF PHLEBOTOMINE SAND FLIES IN SOUTHERN BRAZIL REINHOLD-CASTRO, K.R.; GASPAROTTO, J.C.; NEITZKE-ABREU, H.C.; TEODORO, U. UNIVERSIDADE ESTADUAL DE MARINGÁ, MARINGÁ, PR, BRAZIL.

Keyword:leishmania; breeding sites; sand fly

Abstract: Despite the importance of phebotomine sand flies as vectors of Leishmania, the information about the ecology of immature stages of these insects is rare. The difficulties in finding sand fly larvae and pupae, which develop in wet soils with available organic matter, lead to this lack. The objective of this study, conducted in the Recanto Marista, municipality of Doutor Camargo, Paraná state, was found natural breeding sites of sand flies, since that location has appropriate characteristics for this insects’development. In the first method employed, 94 soil samples were taken from hen houses, tree bases, between tree roots, and under fallen logs. The samples were placed in non-toxic plastic pots (polypropylene) and were kept in a B.O.D. incubator during 4660 days, between 25.5 and 26.5°C, with air relative humidity between 70-90%. The samples were collected from May 2010 to December 2011. In the second method was used a modified emergence trap by Alencar , which had two main parts: 1) the lower, made from metal frame and semitransparent nylon screen, with pyramidal structure (50 by 50 cm on the basis), truncated apex (10 cm diameter) and 45 cm height; and 2) the upper part, formed by an apparatus comprising a funnel and a collector pot (18 cm height), in which was alcohol 70% to preserve the emerged insects. Ten traps were installed 5 to 20 meters from each other, from May 2011 to August 2012 in a remaining forest area significantly altered. These traps were inspected at intervals of 15 to 30 days, to collect sand flies. From incubated soil samples, 21 sand flies emerged and, from emergency traps, eight sand flies. The following species were collected: one specimen of Brumptomyia brumpti, one of Micropygomyia ferreirana, one of Migonemyia bursiformis, one of Migonemyia migonei, one of Pintomyia pessoai, nine specimens of Nyssomyia whitmani, five of Nyssomyia neivai, two of Nyssomyia sp., and five specimens were not identified. It is noteworthy that this is the second finding of Mi. bursiformis in Paraná state.

The collected species have been reported in several endemic areas of cutaneous leishmaniasis in Paraná, including the Recanto Marista, where Ny. whitmani and Ny. neivai have high numerical representation.

- RESERVOIRS [1035] P 850 - SMALL MAMMALS AS HOSTS OF LEISHMANIA SPP. IN AN AREA OF LEISHMANIASIS ENDEMICITY IN NORTH-EASTERN BRAZIL LIMA, B.S.; DE CARVALHO, M.R.; MARINHO JUNIOR, J.F.; DE ALMEIDA, E.L.; BRITO, M.E.F.; GOMES, F.; TORRES, F.D.; XAVIER, E.M.; BRANDÃO FILHO, S.P. CENTRO DE PESQUISAS AGGEU MAGALHÃES, RECIFE, PE, BRAZIL.

Keyword:reservoirs; leishmania braziliensis; leishmania infantum

Abstract: The leishmaniases are parasitic diseases caused by species of Leishmania, whose hosts may be a variety of wild, synanthropic, and domestic animals. The objective of this study was to identify the species of Leishmania found in small mammals in an area in north-eastern Brazil where visceral and cutaneous leishmaniases occur in sympatry. A total of 180 wild and synanthropic animals trapped from 2003 to 2006 in a rural area located in the north part of the Atlantic Forest region of Pernambuco State were included in this study. Samples were assessed by two PCR protocols, one targeting the kinetoplast DNA (kDNA) of Leishmania species belonging to the subgenus Viannia and other the kDNA of Leishmania (Leishmania) infantum and through the immunochromatographic test with rK39 (recombinant antigen of L. infantum). Forty animals were positive to Leishmania (Viannia) and 14 to L. infantum. Five animals from four different species (Didelphis albventris, Nectomys squamipes, Rattus rattus and Holochilus scieurus) were positive by both PCR protocols, suggesting co-infection by L. (L.) infantum and Leishmania (Viannia) braziliensis. A percentage of 5% positivity was found by immunochomatography whith rK39 recombinant. Remarkably, an isolate obtained from a South American water rat (N. squamipes) was characterized by monoclonal antibodies and multilocus enzyme electrophoresis as L. (V.) braziliensis. These results suggest the role of small mammals (rodents and marsupials) as hosts of L. (V.) braziliensis in the study area. Importantly, our findings add weight to the hypothesis about the existence of a sylvatic cycle of L. (L.) infantum, with the involvement of rodent species as hosts.

- EPIDEMIOLOGY [1036] P 851 - CHARACTERISTICS OF AMERICAN TEGUMENTARY LEISHMANIASIS IN REFERENCE UNIT IN MATO GROSSO DO SUL STATE, BRAZIL HIGA JÚNIOR, M.G.; PANIAGO, A.M.M.; DRUZIAN, A.F.; OLIVEIRA, S.M.D.V.L.; FRANÇA, A.N.; NUNES, J.O.; GUIMARAES, L.N.A.; MOURA JUNIOR, I.L.; SOUZA, A.C.N.; DORVAL, M.E.M.C. UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL, CAMPO GRANDE, MS, BRAZIL.

Keyword:leishmaniasis; epidemiology; mato grosso do sul

Abstract: Introduction: The American Tegumentary Leishmaniasis (LTA) is a zoonotic disease that affects humans, some species of wild and domestics animals. The cutaneos form located is characterized for ulcers and painless lesions; the mucous form is characterized for lesions that affect nasopharyngeal regions; the disseminated form presents multiple skin ulcers by hematogenous or lymphatic dissemination; the diffuse form presents nodular lesions without ulcerations. LTA has been afflicting human beings for many centuries, and, in Brazil, in the past three decades, there has been an important increase in the number of cases and also in its geographical distribution. Many cases are registered in Mato Grosso of Sul. Materials and Methods: This work was carried out through analysis of LTA medical records and Chips of Epidemiological Research of the Hospital Epidemiology Service, generated from visits to the

hospital and to the outpatient care in 2011 and 2012. Results: In these years, 51 new cases were recorded in Reference Center for Infectious Diseases of Campo Grande, Mato Grosso do Sul, Brazil. 58,2% (30/51) were male. The cutaneous form occurred in 82,3% (42/51) and the mucous form in 17,6% (9/51). Of these, 7,8% (4/51) were relapse. 88,2% (45/51) of cases were from Mato Grosso do Sul, 1,9% (1/51) of Mato Grosso, 1,9% (1/51) of Amazonas and 7,8% (4/51) were indeterminate. The co-infection LTA-HIV/AIDS were described in 5,8% (3/51).

The cases were confirmed by laboratory criteria in 98,0% (50/51) and 1,9% (1/51) by clinicepidemiological criteria. Glucantime 68,6% (35/51) was the first choice on treatment, followed

by Amphotericin B 13,72% (7/51) and Lipossomal Amphotericin 5,8% (3/51) and 11,7% (6/51) cases not informed. Conclusion: The patients infected by LTA in this service were generally male, developed the cutaneous form, came from Mato Grosso do Sul, were not co-infected with HIV/AIDS and most showed no relapses. Only in one case the diagnostic criteria was not laboratory and the drug generally used was glucantime.

- EPIDEMIOLOGY [1038] P 852 - DOMESTIC ANIMALS AS RISK FACTORS FOR VISCERAL LEISHMANIASIS IN NORTHEASTERN BRAZIL GOMES, A.A.D.1; SILVA, A.R.S.2; LAURENTI, M.D.3; LEUTENEGGER, C.4; ESTRADA, M.4; VIEIRA, R.F.C.5; FOLEY, J.E.6; BIONDO, A.W.7; MARCONDES, M.8 1.UNIVERSIDADE FEDERAL DO VALE DO SÃO FRANCISCO, PETROLINA, PE, BRAZIL; 2.SÃO PAULO STATE UNIVERSITY, BOTUCATU, SP, BRAZIL; 3.UNIVERSITY OF SÃO PAULO, SÃO PAULO, SP, BRAZIL; 4.IDEXX LABORATORIES, WEST SACRAMENTO, UNITED STATES; 5.UNIVERSIDADE FEDERAL DA PARAÍBA, AREIA, PB, BRAZIL; 6.UNIVERSITY OF CALIFORNIA, DAVIS, UNITED STATES; 7.UNIVERSIDADE FEDERAL DO PARANÁ, CURITIBA, PR, BRAZIL; 8.SCHOOL OF VETERINARY MEDICINE, SÃO PAULO STATE UNIVERSITY, ARACATUBA, SP, BRAZIL.

Keyword:cats; dogs; leishmania infantum chagasi

Abstract: The aim of this study was to investigate domestic animals as risk factors for visceral leishmaniasis in the city of Juazeiro, Bahia State, northeastern Brazil. Epidemiological data from 21 cases of Human Visceral Leishmaniasis (HVL) that occurred between 2007 and 2010 were used. The households from each HVL case were identified. Households located within a 250 m perimeter of each household in which HVL cases occurred were visited, and those having domestic animals were included in the study, totalizing 298 households. Blood samples were collected from dogs (n=383), cats (n=84), goats (n=15), donkeys (n=9), sheep (n=4) and horses (n=3), and tested for Leishmania infantum chagasi by enzyme linked immunosorbent assay (ELISA) and real-time PCR (qPCR). A total of 30/498 (6.02%) domestic animals were seropositive for L. infantum chagasi, 20/383 (5.22%) dogs and 10/84 (11.9%) cats. Two (0.4%) domestic animals tested positive by qPCR, both were dogs which were seronegative for L. infantum chagasi. All goats, sheep, horses and donkeys samples tested negative for Leishmania sp. by ELISA and qPCR. Cats were more likely to be positive for L. infantum chagasi than dogs (OR=2.21; 95% CI, 1.1 – 4.81; p=0.043), and domiciliated cats were more likely to be positive for L. infantum chagasi than semi-domiciliated (OR=9.57; 95% CI, 2.21 – 41.37; p=0.0006).

Living next to a HVL case was not associated with positivity to L. infantum chagasi (p=0.5895).

Households presenting > 1 animal were more likely to present positivity to L. infantum chagasi than those presenting 1 animal (OR=3.46; 95% CI, 1.49 – 8.06; p=0.0025). These results highlight the importance of cats as reservoir of L. infantum chagasi.

- EPIDEMIOLOGY [1040] P 853 - EPIDEMIOLOGICAL ASPECTS OF VISCERAL LEISHMANIASIS IN THE MUNICIPALITY OF CAUCAIA, CEARÁ, BRAZIL, 2001-2011 MARTINS-MELO, F.R.; LIMA, M.S.; ALENCAR, C.H.M.; RAMOS JÚNIOR, A.N.; HEUKELBACH, J.

UNIVERSIDADE FEDERAL DO CEARÁ, FORTALEZA, CE, BRAZIL.

Keyword:visceral leishmaniasis; epidemiology; transmission

Abstract: Visceral leishmaniasis (VL) in Brazil is presented by increasing severity of clinical disease, geographic expansion and urbanization. Caucaia (325,441 inhabitants) in the Metropolitan Region of Fortaleza in Ceará state (Northeast of Brazil), is classified as a municipality of intense transmission and a thus focus area for surveillance actions and control of VL. Here we describe the epidemiological characteristics of VL in Caucaia, from 2001 to 2011. We performed a study with secondary data from the Information System for Notifiable Diseases (Sistema de Informação de Agravos de Notificação – SINAN). We used all cases of VL in residents of the municipality of Caucaia (2001-2011) to describe the epidemiological characteristics, using the indicators: incidence rate (per 100,000 inhabitants) and case fatality rate. In both indicators we analyzed temporal trends by simple linear regression. A total of 262 cases and 16 deaths due to VL were reported in the municipality in the study period. The mean incidence rate was 7.6 cases per 100,000 inhabitants (ranging from 4.6 to 12.6 cases per 100,000). Mean case fatality rate was 6.1% (ranging from 0 to 11.9%). The cases were predominantly of male gender (63.7%), children <15 years of age (48.4%), brown race/color (64.5%), and mostly autochthonous of the municipality (93.0%) with the majority of patients living in the urban area (85.5%). The diagnosis was confirmed by laboratory tests in 95.9%. Cure rate was 85.9%. Leishmania/HIV co-infection was recorded in 4.65% cases. The incidence rate (R2: 0.649, p=0.003) and case fatality rate by VL (R2: 0.496, p=0.016) presented significant increase in the period. In the municipality of Caucaia, VL is common, with intense active transmission. Control strategies have not presented sufficient effectiveness to reduce incidence. We emphasize that surveillance actions and control should be prioritized and implemented in an integrated manner.

- EPIDEMIOLOGY [1045] P 854 - OCCURRENCE OF SEROPOSITIVITY FOR LEISHMANIA INFANTUM CHAGASI IN DOGS FROM BUILT-UP AREAS OF THE CITY OF PORTO ALEGRE, RIO GRANDE DO SUL-BRAZIL TEIXEIRA, M.C.1; FREITAS, T.D.1; BISOL, J.1; ROCHA, A.G.1; TARTAROTTI, A.L.2; RAMOS, R.3; STOBBE, N.S.4; ARAUJO, F.A.P.1 1.UFRGS, PORTO ALEGRE, RS, BRAZIL; 2.CEVS/SS/POA, PORTO ALEGRE, RS, BRAZIL; 3.IPB/LACEN, PORTO ALEGRE, RS, BRAZIL; 4.ICBS/UFRGS, PORTO ALEGRE, RS, BRAZIL.

Keyword:canine visceral leishmaniasis; seroepidemiology; rapid test tr-dpp

Abstract: Visceral leishmaniasis is an infectious disease of chronic character whose etiologic agent in Brazil is the protozoan Leishmania infantum chagasi. Dogs are considered domestic reservoirs of such agent. Up to December 2008, the state of Rio Grande do Sul was considered a harmless area for human and canine visceral leishmaniasis. From 2009, autochthonous cases of the disease in human beings and dogs have been confirmed by the State Health Surveillance Center in São Borja, a city in the western border with Argentina. Between 2010 and 2012, the municipalities of Santa Cruz do Sul, Uruguaiana and Viamão also presented cases of canine visceral leishmaniasis (CVL). In Porto Alegre, the State capital, the first case of CVL was notified in 2010 and confirmed by ELISA test and the growth of the protozoan. The city is characterized for being a transitional zone of forest and has borders with the municipality of Viamão. On this condition, the capital was considered to be an area with cases of the disease. In order to know the occurrence of seropositive dogs in Porto Alegre/RS, 245 canine serum samples will be collected. Up to the present moment, 140 canine serum samples have been collected and analyzed in three communities of built-up areas of the city. These communities

have in common the fact of being relocated from peri-urban to central areas. They are socially vulnerable communities whose main income comes from collection and recycling of waste. At the time of sample collection, animal owners are informed about the purpose of this epidemiological research and respond a questionnaire. In addition, they sign a consent form to participate in the study. Serum samples are tested through TR-DPP rapid test kit (Biomangunhos®) which uses the recombinant antigen K28 of Leishmania infantum chagasi. So far, three animals have showed positive reaction in the test, indicating the occurrence of 2.1% (3/140). All samples will subsequently be tested by ELISA technique. Although the research is still in progress it is possible to affirm the occurrence of CVL cases in the capital city. Thus, it is necessary to increase public health professionals’ awareness, including veterinarians, as well as to develop clarification campaigns for the lay population regarding the risks of the disease and its prophylactic measures.

- RESERVOIRS [1048] P 855 - PROSPECTIVE STUDY ON THE DYNAMIC OF THE CLINICALIMMUNOLOGICAL EVOLUTION OF CANINE LEISHMANIA (LEISHMANIA) INFANTUM CHAGASI-INFECTION IN AMAZONIAN BRAZIL CARNEIRO, L.A.1; LIMA, L.D.R.1; DE JESUS, R.C.S.1; LAURENTI, M.D.2; SILVEIRA, F.T.1 1.EVANDRO CHAGAS, BELEM, PA, BRAZIL; 2.UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL.

Keyword:prospective study; canine infection; amazonian brazil

Abstract: Background: There are few reports concerning the evolution of canine visceral leishmaniasis in Brazil, where the euthanasia of seropositive dogs is recommended by the Health Ministry as a control measure. This study has the main objective to characterize the dynamic of the clinical and immunological evolution of canine Leishmania (Leishmania) infantum chagasi-infection from endemic area of AVL. Methods: The prevalence and incidence (6 and 12 moths) of a cohort of 320 mongrel dogs from Barcarena municipality, Pará state, Brazil, were evaluated by IFAT-IgG and DTH, providing the clinical-immunological profile of canine L. (L.) infantum chagasi-infection.

Results: The IFAT and DTH reactivity recognized three different profiles: (1) IFAT+/DTH(n=104), (2) IFAT-/DTH+ (n=18), and (3) IFAT+/DTH+ (n=14), revealing an infection prevalence of 42.5% (138/320). Thirty percent of dogs from the profile 1 presented clinical signs of disease and 100% of dogs from profile 2 were asymptomatic. Concerning the evolution of the profile 1, most of dogs increased the serological titles, presented positive parasitological diagnosis and naturally died, confirming the high susceptibility of L. (L.) infantum chagasiinfection. However, 22.2% of dogs in the profile 2 maintained asymptomatic with negative serology showing a resistant profile of infection. The evolution of the profile 3, 62.5% of dogs maintained slight to moderate serological title and 37.5% showed increased of the serological titles conferring the susceptibility profile to this group.

Conclusion: These results confirm the high susceptibility of dogs to L. (L.) i. chagasi-infection, which contrasts to the human infection that presents a high predominance of resistance against the infection in endemic area.

- RESERVOIRS [1049]

P 856 - VISCERAL LEISHMANIASIS IN DOGS EXAMINED AT VETERINARY CLINICS FROM THE MUNICIPALITY OF ARAGUAÍNA, TOCANTINS, BRAZIL. SANTOS, H.D.1; GALVÃO, S.R.1; MINHARRO, S.1; BAILONA, G.H.C.1; GEORGETTI, E.D.S.1; CASTRO, A.A.P.2; DIAS, F.E.F.1; CAVALCANTE, T.V.1; NEGREIROS FILHO, O.3 1.UNIVERSIDADE FEDERAL DO TOCANTINS, ARAGUAINA, TO, BRAZIL; 2.FUNDAÇÃO DE MEDICINA TROPICAL DO TOCANTINS, ARAGUAINA, TO, BRAZIL; 3.SECRETARIA MUNICIPAL DE SAÚDE, ARAGUAINA, TO, BRAZIL.

Keyword:frequency; amastigotes; clinical signs

Abstract: Since 2003 the state of Tocantins has recorded the highest incidences of Human Visceral Leishmaniasis in Brazil, reaching 33.9/100.000 inhabitants in 2008 and is also considered an intense transmission area. Canines surveys and euthanasia of seropositive dogs are often carried out by public health agencies using serological methods for diagnosis as recommended by the Brazilian Ministry of Health. Due to the volume of samples to be processed by the official laboratories, the time between sample collection and removal of positive households reaches up to 60 days, which is one factor that may contribute to the lack of success in controlling the disease. The purpose of this study was to determine the frequency of Leishmania in dogs sent to Araguaína city veterinary clinics, using the direct detection of parasites as diagnostic test. The study population were 468 dogs of different breeds and ages attended at veterinary clinics in the city of Araguaína in 2007 and underwent clinical examination, and after this a sample of lymph node cells by needle aspiration were collected. The material was deposited on a microscope slide, stained and stretched using panotico kit. In 187 (40%) dogs were found amastigotes consistent with Leishmania and 281(60%) were negative. 133 (71.1%) animals positives showed clinical signs of the disease and 54 (28.9%) were considered asymptomatic. The parasitological examination, despite having low sensitivity is a low-cost, quick and easy implementation test, which can help to reduce the residence time of the reservoirs in the home environment, reducing the risk of human infection. The results demonstrate the needs for continued vigilance by the owners of the animals in relation to leishmaniasis in areas of intense transmission, and the intensification of veterinary surveillance to find positive animals as soon as posible.

- OPERATIONAL HEALTH [1050] P 857 - PUBLIC HEALTH LABORATORY AND EPIDEMIOLOGIC SURVEILLANCE: WORKING IN ACTIVE SEARCH AND EARLY DIAGNOSIS OF HUMAN VISCERAL LEISHMANIASIS BERTOLLO, D.M.B.1; GOMES, É.V.R.1; FIGUEIREDO, J.K.1; PEDRO, N.F.1; SOARES, M.M.C.N.1; ZINI, R.M.1; HIRAMOTO, R.M.2; BOCCHI, M.R.3; TOLEZANO, J.E.2

1.CENTRO DE LABORATÓRIO REGIONAL INSTITUTO ADOLFO LUTZ SÃO JOSÉ DO RIO PRETO - X, SÃO JOSÉ DO RIO PRETO, SP, BRAZIL; 2.INSTITUTO ADOLFO LUTZ - CENTRO DE PARASITOLOGIA E MICOLOGIA - SÃO PAULO, SÃO PAULO, SP, BRAZIL; 3.GRUPO DE VIGILÂNCIA EPIDEMIOLÓGICA 29, SÃO JOSÉ DO RIO PRETO, SP, BRAZIL.

Keyword:american visceral leishmaniasis; public health laboratory; epidemiologic surveillance

Abstract: American visceral leishmaniasis (AVL) in Latin America is caused by Leishmania chagasi its transmission occurs mainly by Lutzomyia longipalpis. It is a zoonosis in expansion in Brazil and is leaving the rural area and becoming increasingly urbanized. Dogs are the major reservoir of the parasite and play a central role in the cycle of transmission to humans. AVL is endemic in rural areas of most Brazilian states and is considered today as a reemerging infection due to its rapid expansion into non-endemic areas. This work presents the important role of public health laboratory with epidemiological surveillance for early diagnosis of LVA in São José do Rio Preto region, State of São Paulo. Were analyzed the database Information System Disease Surveillance (SINAN) and Integrated Management System Hospital (SIGH) used at Regional

Laboratory Center Institute Adolfo Lutz Sao Jose do Rio Preto - X, laboratory conducting the parasitological examination, rapid test and serological, in the period from 2008 to 2012. Since the first confirmed human case of AVL in 2008, died, in the city of Jales, located northwest of São Paulo in the Region of São José do Rio Preto, the disease has shown broad expansion in the municipalities of the region, from just one municipality in 2008 for 5 with occurrence of human cases in 2012. The attention of health services and surveillance aimed at effectively search for cases of rapid testing and deployment from 2011, resulted in an increase of more than 70% of suspected cases, which resulted in greater numbers of confirmed cases (20%). Regarding the occurrence of deaths, in 2008 there was 60% of deaths (3/5), but in the years following significant falls. The action of Epidemiologic Surveillance in conjunction with Public Health Laboratory, has collaborated with the diagnosis faster and more efficient, which promotes early treatment resulting in better prognosis for the patient.

- CELLULAR AND MOLECULAR BIOLOGY [1054] P 858 - NATURAL INFECTION OF NYSSOMYIA NEIVAI BY LEISHMANIA (VIANNIA) IN ISLANDS OF PARANÁ RIVER, SOUTHERN BRAZIL REINHOLD-CASTRO, K.R.1; SANTOS, B.A.1; CRISTOVÃO, E.C.2; GASPAROTTO, J.C.1; RANUCCI, T.R.1; NEITZKE-ABREU, H.C.1; TEODORO, U.1 1.UNIVERSIDADE ESTADUAL DE MARINGÁ, MARINGÁ, PR, BRAZIL; 2.NÚCLEO DE ENTOMOLOGIA, PORTO RICO, PR, BRAZIL.

Keyword:leishmania; nyssomyia neivai; natural infection

Abstract: The technique of multiplex polymerase chain reaction (multiplex PCR) has been useful in the diagnosis of leishmaniasis and detection of Leishmania infection in sandflies and reservoirs, contributing to the knowledge of the disease epidemiology. The aim of this study was to investigate the natural Leishmania infection in sand flies in Mutum, Bandeira, Longa, and Carioca islands of Paraná river, northwest of Paraná state, using multiplex PCR. Part of the native forest is preserved, with low degree of degradation, where small wild mammals live, which are likely natural Leishmania reservoirs. Those islands inserted in the Environmental Protection Area (APA) of Islands and Lowlands of Paraná river. Sand flies were collected in the islands in June (18:00 to 06:00) and August 2012 (18:00 to 00:00), with Falcão light traps. The sand flies were identified and conserved in tubes containing isopropanol, in pools with 9 to10 females. DNA extraction was carried out with a guanidine isothiocyanate and phenol solution.

Two pairs of primers were used, MP3H and MP1L, to amplify a fragment of 70-pb of the conserved region from the minicircle of the kinetoplast of the subgenus Leishmania (Viannia) and 5Llcac and 3Llca to amplify a fragment of 220-pb from the IVS6 gene region of the cacophony in insects of the genus Lutzomyia. The amplification products were analyzed electrophoresis in agarose gel, and the presence of bands was observed in transilluminator. Were identified 529 sand flies female of the Nyssomyia neivai species. A total of 378 females (38 pools) were tested by multiplex PCR. All pools showed the 220-bp fragment for sand flies PCR detection indicating the absence of inhibitors of Taq DNA polymerase. Taking into consideration that one sandfly was infected in each pool, the infection rate found in multiplex PCR was 0.26%, i.e., at least one female of Ny. neivai from the Mutum island showed DNA fragment of Leishmania (Viannia).

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1057] P 859 - WARILEYA ROTUNDIPENNIS (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) POTENCIAL VECTOR DE LEISHMANIA PANAMENSIS?

FERRO, C.; DELGADO, M.; MARIN, D.; RUBIANO, L.C.; COSSIO, A.; ROSALES, M.; MORENO, M.; OCAMPO, C.; GORE SARAVIA, N. CIDEIM, CALI, COLOMBIA.

Keyword:warileya rotundipennis; vector; leishmania

Abstract: Introduction: Various species of the subfamily Phlebotominae have been incriminated as vectors in the transmission of cutaneous leishmaniasis. Entomologic studies conducted in the community La Cabaña, Department of Risaralda within the Pacific coast región of Colombia identified 5 species of the subfamily Phlebotominae, within households. The most abundant was Warileya rotundipennis followed by Psychodopygus panamensis, Nyssomyia trapidoi, Lutzomyia hartmanni y Lutzomyia lichyi. Although this species has not implicated in transmission of leishmaniasis, considering that the prevalence of infection in this community is 85% the intradomicilary high abundance of Warileya, we conducted analyses of the source of blood meals and infection status of the engorged specimens of Warileya rotundipennis.

Methods: Sand flies were collected using CDC light traps in the bedrooms of the households from 6 pm to 6 am during two consecutive days in each of all 9 households of La Cabaña in August of 2011. Sand flies were identified taxonomically to the species level and blood source was determined by amplification and sequencing of the cytochrome b gene. Infection was evaluated by amplification of Leishmania (Viannia) kDNA and qRTPCR of 7SLRNA gene sequences.

Results: DNA amplified from 32/36 specimens was adequate for sequencing, which revealed that the bloodmeals of 27 female Warileya (84%) were of human origin, while one was from a sloth (Choloeopus didactylus), a known reservoir of Leishmania Viannia subspecies, and 4 were not identifiable. Leishmania specific sequences of kDNA and 7SLRNA were amplified from the specimen that had fed on the sloth. Melting peak data from the 7SLRNA qPCR product and sequencing corroborated the origin of the amplified product as Leishmania and the melting temperature coincided with that of the L.panamensis control.

Conclusiones: The possibility that Warileya rotundipennis may participate in transmission of leishmaniasis in La Cabaña is suggested by these findings but will require further investigation including detection of infection in flies that are not blood-engorged since the bloodmeal itself could harbor infection

- CONTROL PROGRAMS [1058] P 860 - CHANGING OF THE PROTOCOL OF LABORATORY DIAGNOSIS OF CANINE VISCERAL LEISHMANIASIS IN PERNAMBUCO, 2012.

DA SILVA, A.1; CABRAL DE FRANÇA, T.M.1; DA SILVA, J.C.1; SILVA, M.D.C.F.1; DE FRANÇA MENDES, F.G.1; DE CARVALHO, T.G.2; AGUIAR, V.B.2; DO NASCIMENTO, L.A.2; DE OLIVEIRA, G.M.1 1.LACEN PE, RECIFE, PE, BRAZIL; 2.SES PE, RECIFE, PE, BRAZIL.

Keyword:canine visceral leishmaniasis; immunoassay; laboratory training

Abstract: INTRODUCTION: In compliance with the determination of the Ministry of Health regarding the change of protocol for the laboratory diagnosis of Canine Visceral Leishmaniasis (CVL) (Technical Note joint Nº 01/2011 "Clarifications on replacing the protocol diagnosis of canine

visceral leishmaniasis" - CGDT - / CGLAB / DEVIT / SVS / MS), the Central Laboratory of Pernambuco (Lacen-PE), in partnership with the Environmental Monitoring of Pernambuco, trained technicians in the municipalities of the State responsible for the collection of blood in dogs for Rapid Test Platform Double route of Canine Visceral Leishmaniasis (RTPDR/CVL).

OBJECTIVE: To present the results of the trainings conducted by Lacen-PE for use of RTPDR/CVL in Pernambuco, showing the number of trained professionals and municipalities.

MATERIAL AND METHODS: The trainings were conducted in the twelve Regional Health districts that comprise the state of Pernambuco and the dependencies of Lacen-PE, with theoretical and practical classes during the year 2012. RESULTS: We conducted 71 training courses, involving 168 professionals from 72 (39%) of the counties among the 184 existing in Pernambuco. These courses were distributed in the 12 Regional Health districts in the state.

CONCLUSION: The RTPDR/CVL is being used throughout the state of Pernambuco and Lacen-PE is responsible for quality control of the results, observing a good performance test with a high degree of agreement among the analyzes, confirming the learning promoted training, which ensures a reliable diagnosis that is rapid and accurate. Thereby, contributing to the effective control of the disease.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1061] P 861 - STUDY VECTORIAL CAPACITY OF PINTOMYIA FISCHERI (PINTO) (DIPTERA: PSYCHODIDAE) TO LEISHMANIA (VIANNIA) BRAZILIENSIS VIANNA DINIZ, M.M.C.S.L.; LAVITSCHKA, C.O.; OVALLOS, F.G.; OLIVEIRA, E.F.; GALATI, E.A.B. FACULDADE DE SAÚDE PÚBLICA - USP, SÃO PAULO, SP, BRAZIL.

Keyword:vectorial capacity; phlebotomines; american cutaneous leishmaniasis

Abstract: Introduction American cutaneous leishmaniasis (ACL) is an anthroponosis involving several species of Leishmania, mammalian reservoirs and phlebotomine vectors. Objective To identify the vectorial capacity of Pintomyia fischeri for Leishmania (Viannia) braziliensis comparing it with that of Nyssomyia intermedial. Methods The following parameters were identified: in the field – density of the species in relation to a host (hamster), with the installation of a Disney trap in two areas: Embu municipality and Iporanga municipality, and a new trap (Iporanga); in the laboratory – on the basis of the experimental infection of sandflies which fed on hamsters infected with the parasite; daily probability of survival of the sandflies; duration of the gonotrophic cycle; extrinsic period of incubation of the parasite; proportion of phlebotomines which fed on the infective host and which themselves became infectious. With these data the vectorial capacity was calculated, using the expression V = m.a.b.E. Results As regards the parameter observed in the field density (m) of in the Disney trap in Iporanga this value for Ny. intermedia was 0.027 and for Pi. fischeri, 0.011, in Embu, for Pi. fischeri , 0.103 and for the new trap in Iporanga, for Ny. intermedia, 7.9 and for Pi. fischeri, 0.11. In the laboratory: infective life expectancy : Ny. intermedia (0.84 days) and Pi. fischeri (0.89 days); proportion of females fed on the hamster: Ny. intermedia 0.67 and Pi. fischeri 0.77 and median of the duration of the gonotrophic cycle for both species, of 5 days; and proportion of females infected which attained infective form (b) for Ny. intermedia 0.90 and Pi. fischeri 0.59. The vectorial capacity obtained with the Disney for Ny. intermedia in Iporanga was 0.0027 and for Pi. fischeri, 0.00089 and in Embu for Pi. fischeri, 0.0083 and for the new trap only in Iporanga, Ny. intermedia 0.8 and Pi. fischeri 0.0089. Conclusion The vectorial capacity measured by the new and the Disney traps presented considerable differences. For Ny. intermedia in Iporanga the value of the former was 296.3 times greater than that of the Disney and for Pintomyia fischeri it was 10 times greater. In the comparison between traps of the same kind, in Iporanga for Ny. intermedia the value of vectorial capacity obtained with the Disney trap was 3.03 times greater than that of Pi. fischeri and in the new trap it was 89.9 times greater. For Pintomyia fischeri, the vectorial capacity with Disney (Embu) was 9.3 times greater than that for Iporanga. The vectorial capacity of Pi.

fischeri (Embu) was 3.07 times greater than that of Ny. intermedia (Iporanga). Possibly both species are active in the transmission of ACL in the state of São Paulo. In Iporanga Ny. intermedia and Embu Pi. fischeri. Financial support: FAPESP. - CELLULAR AND MOLECULAR BIOLOGY [1075] P 862 - INVESTIGATION OF NATURAL INFECTION OF SAND FLIES BY LEISHMANIA (VIANNIA) IN PARANÁ STATE, SOUTHERN BRAZIL REINHOLD-CASTRO, K.R.; RANUCCI, T.R.; NEITZKE-ABREU, H.C.; GASPAROTTO, J.C.; SANTOS, B.A.; TEODORO, U. UNIVERSIDADE ESTADUAL DE MARINGÁ, MARINGÁ, PR, BRAZIL.

Keyword:leishmania; sand fly; natural infection

Abstract: Leishmaniasis is widespread in the world and has a great impact on public health. The life cycle of the parasites that cause leishmaniasis involves sand fly vectors and mammalian reservoirs that sustain the life cycle of Leishmania in rural and urban areas. The multiplex polymerase chain reaction (multiplex PCR) is a technique with high specificity and sensitivity and has been useful for detecting the presence of Leishmania in sand flies. The objective of this study was to investigate the natural Leishmania infection in sand flies in Bandeirantes municipality, Paraná state. Sand fly collections were conducted in a rural area called Cateto Nomura, in March, April, May, August and September 2012, between 18:00 and 2:30 hours, with Shannon trap. After identification, the sand flies were conserved in tubes containing isopropanol, in pools with 6 to 10 female. The insects were macerated in lysis buffer and DNA extraction was carried out with a guanidine isothiocyanate and phenol solution. Two pairs of primers were used for multiplex PCR: i) the MP3H and MP1L to amplify a fragment of 70-pb of the conserved region from the minicircle of the kinetoplast of the subgenus Leishmania (Viannia); ii) the Llcac and 3Llca to amplify a fragment of 220-pb from the IVS6 gene region of the cacophony in insects of the genus Lutzomyia. The primers 5Llcac-3Llcac were used with the function of internal control reaction. The amplification products were submitted to electrophoresis in agarose gel, and the presence of bands was observed in transilluminator. We identified 837 sand flies female; of which 679, separated in 69 pools, were tested by multiplex PCR. Were identified specimens of Pintomyia pessoai, Nyssomyia whitmani, Pintomyia fischeri, Nyssomyia neivai, Mi. migonei, Expapillata firmatoi e Brumptomyia brumpti. All 69 pools showed the 220-bp fragment for sand flies DNA detection, but were not found anyone with Leishmania infection. The multiplex PCR has proved effective for detecting Leishmania infection in sand flies. The collected species are among the most frequent in anthropic environments, in Paraná state, but there are a few studies of vector competence in this state. Thus, it is necessary to continue the search for Leishmania infection in sand flies.

- EPIDEMIOLOGY [1077] P 863 - A COMPARATIVE STUDY OF CDC LIGHT TRAPS AND THE EMERGENCE ONES TO THE EPIDEMIOLOGICAL SURVEILLANCE OF PHEBOTOMUS (SANDFLY) PROCEEDING FROM THE ATLANTIC FOREST DA SILVA, D.T.1; PIRAJÁ, G.V.1; C. RODAS, L.A.2; F. CHIERICI, N.2; M. PEREIRA, D.C.2; B. LUCHEIS, S.3; VICTÓRIA, C.1; MODOLO, J.R.1

1.VETERINARY MEDICINE AND ZOOTECHNY COLLEGE - UNESP / FMVZ - DHVSP, BOTUCATU, SP, BRAZIL; 2.LABORATORY OF LEISHMANIASIS - SUCEN, ARAÇATUBA, SP, BRAZIL; 3.SÃO PAULO STATE ‘S AGENCY OF TECHNOLOGY OF AGROBUSINESS - APTA/SAA, BAURU, SP, BRAZIL.

Keyword:leishmaniasis; sandflies; trap

Abstract: Mainly because of being recorded in several regions of the world and for presenting some variations in the epidemiological cycle- such as the species of the etiological agent, the geographic region taken into account, the mammal species exposed to the risk of infection and the presence of Phlebotumus (sandfly) vector, the clinical and epidemiological diversities caused Leishmaniasis to be one of the six major diseases concerning in public health importance . On considering this context, the present essay is intended for a comparative study of two kinds of traps- the CDC light trap (CDC-LT) and the emergence trap (ET) - to study sandflies obtained form a fragment of the atlantic forest situated in the urban surroundings of the city of Marília/SP. In order to achieve this aim the following, were utilized intermediate to September and December of 2012, ten CDC -LT and ten ET, wicth were placed monthly, both types at the same spot in an area distributed through the Municipal woods “ Rangel Pietraróia”. The CDCLT were place rising to 150 centmetres in height, after dusk for three consecutive nights, whereas the ET were put along the CDC ones at the first day and they remained in the soil for 15 days. Subsequently, the material was sent to the Laboratory of Leishmaniasis/SUCEN, in Araçatuba/SP to the procedures of identification according Galatti (2003). By means of the CDC-LT it was possible collect 2 males and 1 female of the species Lutzomyia longipalpis and 4 females of the species Brumptomyia sp. Traps of this model usually utilized in surveillance activities because of their efficiency in capturing adult mosquitoes. There was no Phlebotomus (sandfly) captured by the ET, which can be justified by both reasons – the traps lasted for a shorter period time than that requires to successfully accomplish the whole cycle of the mosquito and the few episodes of rain in the region and the time investigated. In fact, the emergence of sandflies (Phlebotomus) in forests seems to be fundamentally related to high rainfall rates, because there is a considerable increase during the period of high precipitation. As the results obtained show, it was not possible to correlate the site of oviposition with that of capturing adult specimens. Although the production of sandflies had been nil, due to factors previously mentioned, the efficiency of the ET could be verified for many other kinds of arthropods were captures: diptera insects (buts no sandflies), collembola ones, Hymenoptera (wasps and ants), among others.

Acknowledgment: Conselho Nacional de Desenvolvimento Científico e Tecnológico - CNPq and Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP - VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1078] P 864 - STUDY ON SAND FLIES (DIPTERA: PSYCHODIDAE) RESEARCH AND MOLECULAR LEISHMANIA SPP. COMMUNITIES IN LUTZOMYIA FLAVISCUTELLATA IN THE MUNICIPALITY OF QUILOMBOLA INHANGAPI, PARA STATE, BRAZIL CALDEIRA, R.D.1; DAMASCENO, Á.R.1; DA SILVA, A.F.1; PRADO, W.S.1; DE OLIVEIRA, A.P.1; FRANCO, A.1; DA SILVA, L.C.O.1; DE FARIAS, D.M.1; JÚNIOR, F.D.1; MATOS, P.C.M.1; BARATA, I.R.2; SILVEIRA, F.T.2; CAVALCANTE, G.G.1; SCOFIELD, A.1 1.UNIVERSIDADE FEDERAL DO PARÁ, CASTANHAL, PA, BRAZIL; 2.INSETÁRIO DE FLEBOTOMÍNEOS DO LABORATÓRIO DE LEISHMANIOSES DO INSTITUTO EVANDRO CHAGAS, ANANINDEUA, PA, BRAZIL.

Keyword:phlebotomines; lutzomyia; cutaneous leishmaniasis

Abstract: The phlebotomine sandflies (Diptera: Psychodidae) have importance for the man and the animals because they act as natural vectors of etiologic agents of diseases, such as protozoa of the genus Leishmania, causing of the leishmaniases. The main objective of this study was an epidemiological study on the diversity of fauna flebotominica and search to infection by Leishmania spp. in captured vectors in quilombola communities of the municipality of Inhangapi, in the state of Pará, area with transmission of cutaneous Leishmaniasis. HP automatic

light traps were used installed in indoors, peridomicile and adjacent forest, during the period december 2010 to november 2011. The natural infection rate due to Leishmania spp. was studied in Lutzomyia flaviscutellata (65 females) using the polymerase chain reaction technique.

A total of 1118 phlebotomines were captured and distributed in 22 species of the genus Lutzomyia. Females accounted for 51.97% (581/1118) of the total captured and 48.03% (537/1118) were male. Among the species caught L. auraensis was the more found with 34.4% (385/1118), followed by L. antunesi with 24.5% (275/1118). The species found those that present epidemiological importance were Lutzomyia flaviscutellata, natural vector of Leishmania amazonensis, Lutzomyia antunesi, incriminated as a possible vector of Leishmania lindembergi and Lutzomyia complexa, vector of Leishmania braziliensis. By means of PCR was not detected the presence of Leishmania spp. DNA in the females of L. flaviscutellata caught, however, the presence of species L. flaviscutellata, L. antunesi and L. complexa serves to alert about the risk of transmission of cutaneous leishmaniasis in the studied area.

- CONTROL PROGRAMS [1080] P 865 - SURVEILLANCE AND CONTROL OF VISCERAL LEISHMANIASIS IN AREAS AT RISK OF FLOODING IN THE MOSSORO, RIO GRANDE DO NORTE, BRAZIL COSTA, K.F.L.1; AMORIM, C.F.1; SILVA, L.F.1; KAZIMOTO, T.A.1; REGINALDO, M.L.1; ALBANO AMORA, S.S.1; DE SOUZA, C.S.F.2; NOGUEIRA FREITAS, Y.B.1; BARBOSA, S.R.1; CÂMARA, F.V.1; FERNANDES, A.M.M.M.3; ALVES, N.D.1; CARNEIRO FEIJÓ, F.M.1 1.UNIVERSIDADE FEDERAL RURAL DO SEMI-ÁRIDO - UFERSA, MOSSORÓ, RN, BRAZIL; 2.DEPARTAMANETO DE PROTOZOOLOGIA - FUNDAÇÃO OSWALDO CRUZ - FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.SECRETARIA DE VIGILÂNCIA A SAÚDE - PREFEITURA MUNICIPAL DE MOSSORÓ, MOSSORÓ, RN, BRAZIL.

Keyword:epidemiological surveillance; community health agents; endemic

Abstract: The Epidemiological Surveillance (EV) is part of the Control Program of Visceral Leishmaniasis, proposed by the Ministry of Health in Brazil and aims to reduce the rates of mortality and level of morbidity of the disease. Thus, we attempted to check for surveillance and control of human and canine VL held in the city from the work carried out by Community Health Agents (CHA) in areas at risk of flooding, favorable to presence of vector.

Semistructured individual interviews were conducted with 12 CHA three quarters in the city of Mossoro, in northeastern Brazil about knowledge, surveillance and control actions VL developed for them. It was found that 84% (10/12) of CHA unknown epidemiological surveillance measures of VL, 8% (1/12) knows the epidemiological surveillance in the dog and 8% (1/12) knows at least two types of surveillance VL. All interviewed unknown the classification of areas for surveillance and control of VL. In relation to training about VL, 33% (4/12) never received and 67% (8/12) had received training in the form of update courses, and of these, only 8% (1/12) affirmed that this occurs more than once a year, but the answers were not consensus on that frequency. CHA who have received training 42% (5/12) have addressed the VL human and canine and 100% (12/12) said good learning. About active case finding positive human or canine, 42% (5/12) do not perform search neither cases of human nor canine cases, 25% (3/12) conducted only when there is confirmed or suspected cases of human or canine and 17% (2/12) perform active case finding human. Regarding the activities of canine and human surveillance 83% (10/12) did not perform. About health education for VL 58% (10/12) develop with the population. It is noticed that the CHA unknown epidemiological surveillance of the VL, affecting the prevention and control of disease. And that there is an disagreement about the frequency of trainings showing that each unit have a timeline own health and dissociated, does not covering all CHA. Moreover, there are gaps in knowledge of these professionals, for there is not constant training, neglecting VL in these areas. Thus, it is clear the need to invest in the continuing education of CHA about VL, on the goals and measures for prevention and control of disease.

- EPIDEMIOLOGY [1086] P 866 - GENETIC HOMOGENEITY AMONG LEISHMANIA (VIANNA) BRAZILIENSIS IN A REGION NEWLY ENDEMIC FOR CUTANEOUS LEISHMANIASIS: EPIDEMIOLOGICAL APPLICATION OF MULTILOCUS SEQUENCE ANALYSIS MARLOW, M.A.1; BOITÉ, M.C.2; FERREIRA, G.E.M.2; CUPOLILLO, E.2; STEINDEL, M.1 1.LABORATÓRIO DE PROTOZOOLOGIA, UNIVERSIDADE FEDERAL DE SANTA CATARINA, FLORIANÓPOLIS, SC, BRAZIL; 2.LABORATÓRIO DE PESQUISA EM LEISHMANIOSE, FUNDAÇÃO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:leishmania (vianna) braziliensis; south brazil; multilocus sequence analysis

Abstract: Background: Recently over the past decade, cutaneous leishmaniasis (CL) has been emerging in the southern state of Santa Catarina, Brazil. Today, CL is now endemic in the northeastern region of the state and Leishmania (Vianna) braziliensis (LB) has been incriminated as the species mainly responsible for autochthonous transmission. As knowledge on species and strains of CL has important clinical and vaccine development implications, 94 strains of LB from 12 states in Brazil, including 32 isolates from SC, were compared by multilocus sequence analysis (MSLA) to determine the intraspecies genetic variation among the strains and to use the MSLA output to evaluate epidemiological inferences.

Methods: Consensus DNA sequences from six loci (MPI, 6PGD, MDH mt, MDH nc, HSP70, ICD) were obtained in Phred/Phrap/Consed software package. DNA sequences presenting ambiguous sites were phased in DNAsp through PHASE algorithm. Clonal complexes were defined after BURST analysis. Population structure was investigated using STRUCTURE software with sequence types as input and the location as prior information. The most probable number of populations was estimated by calculation of K. Results: Strains were found to involve four clonal complexes (CC), with majority (57.45%, n=54) conforming to CCI and approximately 37% (n=35) presenting singleton profiles. For SC strains, 50.00% (n=16) belonged to CCI, 46.88% (n=15) were singletons, and only one strain pertained to CCIV. Conforming with the BURST analysis, four main populations were identified in the STRUCTURE analysis, with nearly 60% (n=56) belonging to one main population (POP3). With the exception of two strains, nearly all SC strains were found to be from POP3 (93.55%, n=28). Both of the non-POP3 SC strains were imported cases and conformed to POP2. One of these cases was imported from Mato Grosso, for which all strains from this state were found to be from POP2 as well. No association between clinical form (cutaneous/mucosal) and population was observed (Fisher’s exact test, p= 0.4510).

Conclusion: Results suggest the same major LB genotype circulating in Brazil, as defined by MLSA, is responsible for the CL emergence in Santa Catarina, and also provides further evidence of the adaptability of this main genotype to various ecosystems and possibly vectors.

The discriminatory power of these six loci for MLSA demonstrates the potential of this method as an epidemiological tool.

- EPIDEMIOLOGY [1088] P 867 - EPIDEMIOLOGICAL ASPECTS OF CUTANEOUS LEISHMANIASIS AND VISCERAL LEISHMANIASIS AND LEISHMANIA SPP. DETECTION IN DOGS OF MAROON COMMUNITIES IN INHANGAPI MUNICIPALITY, PARÁ STATE, BRAZIL DA SILVA, A.F.1; DAMASCENO, Á.R.1; PRADO, W.S.1; CALDEIRA, R.D.1; SILVA, L.C.S.1; DE

MORAIS, R.1; SOUSA, L.O.1; JÚNIOR, F.D.1; GONÇALVES, F.T.1; SANTOS, R.C.1; SILVEIRA, F.T.2; CAVALCANTE, G.G.1; SCOFIELD, A.1

1.UNIVERSIDADE FEDERAL DO PARÁ, CASTANHAL, PA, BRAZIL; 2.INSETÁRIO DE FLEBOTOMÍNEOS DO LABORATÓRIO DE LEISHMANIOSES DO INSTITUTO EVANDRO CHAGAS, ANANINDEUA, PA, BRAZIL.

Keyword:cutaneous leishmaniasis; serology; pcr

Abstract: The study aimed to contribute to the establishment of the epidemiological profile of cutaneous leishmaniasis and visceral leishmaniasis in dogs of maroon communities in the Inhangapi´s municipality, on Pará, using different diagnostic techniques and different biological samples.

The study was conducted in maroon communities Menino Jesus of Petimandeua and Itaboca, in wich 2011, samples of blood, serum and skin in dogs without injury, also being held to fill a questionnaire containing information about the animals and socio-economic characteristics and cultural inhabitants. Techniques were used indirect ELISA, IFA, imprint, histopathology and PCR for L. i. chagasi, L. amazonensis and complex species L. braziliensis. Of the 143 of the dogs, 2.1% (3/143) were seropositive for Leishmania spp. by ELISA and IFAT and 8.4% (12/143) were PCR positive for L. i. chagasi. The three seropositive dogs were not positive in the PCR's. In positive samples of PCR of L. i. chagasi amplification was detected in 8.4% (12/143) from samples of intact skin and 1.4% (2/143) of blood. No sample on the skin was detected DNA from L. amazonensis and complex species of the L. braziliensis. In the only dog euthanized seropositive DNA was not detectable species of Leishmania. The presence of L. i. chagasi associated with the dogs´dispersion, the environmental characteristics, to human action and inadequate sanitation can encourage the spread of this agent and the occurrence of VL in humans in the maroon communities Menino Jesus of Petimandeua and Itaboca, serving as a warning to health authorities.

- CONTROL PROGRAMS [1089] P 868 - KNOWLEDGE OF COMMUNITY HEALTH WORKERS ABOUT VISCERAL LEISHMANIASIS THAT WORK IN AREAS AT RISK OF FLOODING IN THE CITY OF MOSSORO, RIO GRANDE DO NORTE, BRAZIL COSTA, K.F.L.1; AMORIM, C.F.1; SILVA, L.F.1; KAZIMOTO, T.A.1; REGINALDO, M.L.1; ALBANO AMORA, S.S.1; DE SOUZA, C.S.F.2; NOGUEIRA FREITAS, Y.B.1; BARBOSA, S.R.1; DE MACEDO, L.B.1; FERNANDES, A.M.M.M.3; ALVES, N.D.1; CARNEIRO FEIJÓ, F.M.1 1.UNIVERSIDADE FEDERAL RURAL DO SEMI-ARIDO, MOSSORO, RN, BRAZIL; 2.DEPARTAMENTO DE PROTOZOOLOGIA FUNDAÇÃO OSWALDO CRUZ - FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.SECRETARIA DE VIGILÂNCIA A SAÚDE PREFEITURA MUNICIPAL DE MOSSORÓ, MOSSORO, RN, BRAZIL.

Keyword:health public; health education; anthropozoonosis

Abstract: The Visceral Leishmaniasis (VL) is a anthropozoonosis of great importance to public health, it is fundamental to articulation of various professionals and institutions as a way of disseminating information about the disease and environmental conditions relevant for its control and prevention. Thus, we sought to evaluate the knowledge of Community Health Agents (CHA) about VL, since these professionals perform activities of disease prevention and health promotion. Semistructured interviews were conducted with questions about the VL with 12 of CHA Basic Health Units located in areas at risk of flooding in the Mossoro, in northeastern Brazil. It was noted that all CHA interviewees know VL and 67% (8/12) know that VL can affect man and dog. In relation to human VL 92% (11/12) knew at least one symptom, 75% (9/12) cited blood tests main methods of diagnosis, 8% (1/12) unknown the cure, 67% (8 / 12) unknown the treatment, 75% (9/12) denied the existence of VL vaccine and 25% (3/12) not able to answer. Regarding the transmission, 92% (11/12) reported mosquito bites, but none cited as the sandfly vector. In relation to canine VL, 92% (11/12) knew at least one symptom, 92%

(11/12) cited the blood tests as diagnostic methods, 67% (8/12) ensured there is no treatment, of these 50% (6/12) mentioned euthanasia as the only alternative control and only 25% (3/12) know the vaccine. About the preventive measures of VL, 75% (9/12) know at least one, but 25% (3/12) did not know any. Were mentioned specific preventive measures for man, among them 25% (3/12) said the use of repellent, but 50% (6/12) do not know any specific preventive measure for man. The preventive measures mentioned as specific to canine VL were related to the environment in 17% (2/12) and vaccination 17% (2/12), but most 67% (8/12) do not know how to prevent canine VL. One can see that the greatest gaps in knowledge of CHA found about LV are related to the unawareness of humane treatment and prevention measures specific to the man and the dog.

- EPIDEMIOLOGY [1090] P 869 - THE EXPANSION OF CUTANEOUS LEISHMANIASIS IN SANTA CATARINA, BRAZIL. PORTES, M.G.T.1; CAMARGO-NEVES, V.L.2; MAKOWIEKI, M.E.3; FRANÇA, E.M.O.3

1.SECRETARIA DE DESENVOLVIMENTO REGIONAL JOINVILLE, SECRETARIA DE ESTADO DA SAÚDE DE SANTA CATARINA, JOINVILLE, SC, BRAZIL; 2.SUPERINTENDENCIA DE CONTROLE DE ENDEMIAS, SECRETARIA DE ESTADO DA SAÚDE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 3.DIRETORIA DE VIGILÂNCIA EPIDEMIOLÓGICA, SECRETARIA DE ESTADO DA SAÚDE DE SANTA CATARINA, FLORIANÓPOLIS, SC, BRAZIL.

Keyword:cutaneous leishmaniasis; epidemiological profile; expansion of cl

Abstract: In Brazil the number of cases of cutaneous leishmaniasis (CL) has increased from end of the 1980´s, when a clear change in the epidemiological profile occurred. It went from being a disease that typically occurred in the forest and rural environments to peri urban areas. Those modifications are the results of the anthropic action to new agriculture fronts and the building of new urban centers, which have contributed to the establishment of new transmission's cycles. In southern Brazil the CL was known only in the Ribeira Valley Paranaense located in the Serra do Mar Region. In Santa Catarina the first recorded cases of CL are dated 1987 and until the mid2000s, autochthonous cases were only registered sporadically, in the far western counties of the State. However, since 2005 there has been an increasing number of cases accompanied by an increase in the number of municipalities transmission. In this study we have described epidemiological characteristics of the CL cases from 2001 to 2010. All autochthones cases recorded in the National Notifiable System, have been considered, with date of onset of symptoms in the corresponding year. The variables studied were age, sex, clinical presentation and the municipality of infection. As a result we obtained 423 registered autochthones cases distributed in 31 municipalities of the State. The highest detection rate occurred in 2006 (2.5 cases/100,000 inhab.). 156 cases were registred mainly in four counties: Blumenau, Balneário Camboriú, Itapema and Florianópolis. Regarding the epidemiological profile it was observed that: 7.3% of the cases were children aged <10 years of age, 61.8% male, 88.0% of them lived in urban or peri-urban regions, the prevalent clinical form was the skin and prevalent in 99.0% of cases. Since 2005, the state of Santa Catarina has been experiments the expansion of CL with epidemics outbreaks. The epidemiologic characteristics of the cases lead us to think of a recent transmission. The increase in CL State follows the trend that has been occurring in other regions of Brazil. Relevant factors to explain this new profile of the disease in the establishment of new settlements in Serra do Mar Cost in order to obtain cheap labor and also in other parts of the State due to agricultural expansion.

- EPIDEMIOLOGY [1091] P 870 - VISCERAL LEISHMANIASIS ARRIVES AT RIO GRANDE DO SUL, BRAZIL SEVERO, M.M.S.1; DEBONI, S.C.2; BERCINI, M.A.2; ZWETSCH, A.3; BEHAR, P.R.P.1

1.UFCSPA, PORTO ALEGRE, RS, BRAZIL; 2.CEVS/SES-RS, PORTO ALEGRE, RS, BRAZIL; 3.FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:visceral leishmaniasis ; rio grande do sul ; patients

Abstract: The expansion of visceral leishmaniasis (VL) is caused by the presence and adaptation of the vector Lutzomyia longipalpis to the environment, combined to the circulation of Leishmania infantum chagasi in domestic dogs and to socioeconomic and environmental factors that favor its occurrence. Rio Grande do Sul (RS) enters the scene in November 2008 with the notification of the first case of canine visceral leishmaniasis (CVL) in the city of São Borja, diagnosed in October 2008. In January 2009 it was reported the first human case of VL in that city, with symptoms beginning in late 2008. In light of these findings research regarding the vector confirmed Lutzomyia longipalpis in November 2008. Our purpose is to inform professionals about the current scenario of VL in RS describing clinical features and contributing to increase the degree of early detection and treatment. From January 2009 until December 2012 there were 8 cases of VL, 7 from São Borja City and 1 from Uruguaiana, all confirmed by antibody detection by Immunofluorescence Assay (IFA) and / or parasitologic exam. Both cities share mixed-environment urban / suburban / rural characteristics with the presence of organic matter generated by vegetation and pets. The factors of environmental changes generated in peridomestic areas can be aggravated when there are poor sanitary conditions. The patients' ages ranged from 06 to 76 years, being 7 of them, men. All patients lived at homes with dogs with CVL. The most common symptoms were fever (100%) and asthenia (75%). The most common findings on physical examination were hepatomegaly (87.5%), and splenomegaly (75%).

Laboratory abnormalities included anemia, thrombocytopenia, leukopenia, and elevated aminotransferases, bilirubin and lactate dehydrogenase levels. One patient was diagnosed with HIV (human immunodeficiency virus). Another died before the specific treatment for VL. The 7 patients were treated with liposomal amphotericin B and until the conclusion of this study showed no reactivation of VL. The sporadic occurrence of cases complicates the clinical diagnosis, and the report of the cases could contribute to improve the sensitivity of diagnosis.

Early treatment reduces the suffering and death of patients and is a valuable management tool for the VL Program.

- EPIDEMIOLOGY [1095] P 871 - COINFECTION AIDS AND VISCERAL LEISHMANIASIS IN A REFERRAL HOSPITAL GOES, J.J.; CORTEZ, A.M.C.; CORTEZ, L.R.C.; PEDROSA, C.M.S.; ALVES, W.A.; LESSA, M.M.; GOMES, A.R.F.; FREITAS, I.S. UFAL, MACEIÓ, AL, BRAZIL.

Keyword:coinfection; visceral leishmaniasis ; hiv

Abstract: Parasites of the genus Leishmania are opportunistic infectious agents in patients with immunosuppression, such as in individuals with HIV (Human Immunodeficiency Virus) and AIDS (Acquired Immunodeficiency Syndrome). Because the Northeast Region of Brazil is endemic for both infectious agents, we decided to analyze the clinical and epidemiological characteristics of coinfection Visceral Leishmaniasis (VL) / HIV cases, occurring in the referral hospital Dr. Hélvio Auto ( Maceió - Alagoas), reference in infectious and parasitic diseases for SUS (unified health system) Alagoas. Were admitted 10 cases of coinfection HIV-VL between 2003 and 2012. Equal sex ratio. The mean age was 34.2 years. Six cases were from urban area.

The majority from the coastal region. Regarding disease progression, six recovered for VL and four progressed to death. In relation to the time of HIV diagnosis, patients who died had

serological diagnosis at admission and died shortly thereafter, none received previous treatments for HIV and only one case was treated VL. While the survivors were already on antiretroviral therapy. Of the six survivors, five had positive spinal puncture. Of those who died, two had positive spinal puncture, and in the other two, parasites were found in the peripheral blood. The diagnosis of the diseases are being made so late, not being possible to use the therapy, implying a higher probability of death. We highlight the importance of educational campaigns for health professionals and the general population aiming early diagnosis of both diseases.

- CONTROL PROGRAMS [1101] P 872 - KNOWLEDGE OF COMMUNITY HEALTH AGENTS ABOUT CANINE VISCERAL LEISHMANIASIS IN ENDEMIC AREAS IN THE MOSSORO CITY, RIO GRANDE DO NORTE, BRAZIL COSTA, K.F.L.1; AMORIM, C.F.1; SILVA, L.F.1; KAZIMOTO, T.A.1; REGINALDO, M.L.1; ALBANO AMORA, S.S.1; DE SOUZA, C.S.F.2; NOGUEIRA FREITAS, Y.B.1; BARBOSA, S.R.1; CÂMARA, F.V.1; FERNANDES, A.M.M.M.3; ALVES, N.D.1; CARNEIRO FEIJÓ, F.M.1 1.UNIVERSIDADE FEDERAL RURAL DO SEMI-ÁRIDO - UFERSA, MOSSORÓ, RN, BRAZIL; 2.DEPARTAMENTO DE PROTOZOOLOGIA DA FUNDAÇÃO OSWALDO CRUZ - FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.SECRETARIA DE VIGILÂNCIA A SAÚDE - PREFEITURA MUNICIPAL DE MOSSORÓ, MOSSORÓ, RN, BRAZIL.

Keyword:dogs; zoonosis; disclosure

Abstract: The epidemiological chain of Visceral Leishmaniasis (VL) possesses three major elements: the vector, the domestic reservoir, and the susceptible man. The dog is identified as the major domestic reservoir of the protozoa, and, therefore, it is determinant to maintain the transmission cycle of this disease in the urban environment. Thus, to assess the knowledge of community health agents (CHA) about the Canine Visceral Leishmaniasis (CVL), was performed a semistructured interview with 30 CHA of basic health units of endemic districts of the Mossoró city, Rio Grande do Norte, Brazil. Among the agents that were interviewed, 66.7% (20/30) had received a theoretical capacitation course about CVL, which suggested that they may be able to transmit to the population the information about the disease in these animals. Most of the agents (66.7% or 20/30) know three or more symptoms of the CVL, and 90.0% (27/30) of them know that in this species the diagnosis is based on blood tests, nevertheless, they do not know which type of test should be performed. Moreover, 90.0% (27/30) of CHA knows that CVL has no cure, and 83.3% (25/30) affirm that there is no treatment for this disease and indicates the euthanasia as the only measure to be taken after the definitive diagnosis. Only 46.7% (14/30) of CHA know that exist a vaccine to CVL, what demonstrate a lack of knowledge regarding this important prevention method. Concerning the epidemiological surveillance actions of CVL, only 13.3% (04/30) knows the main methods involved. Therefore, despite of the CHA receiving capacitation and having a basic knowledge about the CVL, it is necessary the development of activities more comprehensive and emphatic about the prevention and control methods of the disease in this species to capacitate professionals that promote health education, to reduce or avoid the spread of focuses of this disease.

- EPIDEMIOLOGY [1105] P 873 - PARACOCCIDIOIDES BRASILIENSIS INFECTION IN DOGS SOROPOSITIVE AND SORONEGATIVE FOR LEISHMANIASIS IN MATO GROSSO DO SUL STATE PETRONI, T.F.1; ZUQUE, M.A.S.2; EMPKE, A.3; ONO, M.A.1 1.UNIVERSIDADE ESTADUAL DE LONDRINA, LONDRINA, PR, BRAZIL; 2.UNIVERSIDADE ESTADUAL PAULISTA, BOTUCATU, SP, BRAZIL; 3.CCZ, TRES LAGOAS, MS, BRAZIL.

Keyword:paracoccidioidomycosis; leishmaniasis; dogs

Abstract: Paracoccidioidomycosis (PCM) is a systemic mycosis caused by the fungus Paracoccidioides brasiliensis. PCM is observed in humans and wild and domestic animals. Dogs can be infected

by P. brasiliensis due to the habit of sniffing and digging the soil, the probable fungus habitat.

Dogs are also the main reservoir of Visceral Leishmaniasis (V.L.). Therefore, the objective of this study was to evaluate the infection by P. brasiliensis in dogs seropositive and seronegative for V.L. in the municipality of Três Lagoas, Mato Grosso do Sul state, Brazil. The 349 serum samples (270 positive and 79 negative for leishmaniasis) were evaluated by Immunodiffusion and ELISA using exoantigen of P. brasiliensis as antigen. The dogs seropositive and seronegative for leishmaniasis showed a positivity of 66% and 30%, respectively in the ELISA with P. brasiliensis exoantigen. In the Immunodiffusion test, it was observed a positivity of 9,3% only in seropositive samples for V.L. These results suggest a possible cross-reactivity or a co-infection by Leishmania and P. brasiliensis.

- OTHER SPECIAL TOPICS [1108] P 874 - KNOWLEDGE OF COMMUNITY HEALTH WORKERS ON THE TRANSMISSION OF VISCERAL LEISHMANIASIS IN AREAS AT RISK OF FLOODING IN NORTHEASTERN BRAZIL AMORIM, C.F.1; KAZIMOTO, T.A.1; DE LIMA COSTA, K.F.1; SILVA, L.F.1; REGINALDO SOUSA, M.L.1; ALBANO AMÓRA, S.S.1; NOGUEIRA FREITAS, Y.B.1; BARBOSA, S.R.1; ALVES, N.D.1; CARNEIRO FEIJÓ, F.M.1; RIBEIRO, T.T.1; FERNANDES, A.M.M.M.2 1.UNIVERSIDADE FEDERAL RURAL DO SEMI-ÁRIDO – UFERSA, MOSSORÓ, RN, BRAZIL; 2.SECRETARIA DE VIGILÂNCIA À SAÚDE – PREFEITURA MUNICIPAL DE MOSSORÓ/RN, MOSSORÓ, RN, BRAZIL.

Keyword:health professionals; sandfly; environment

Abstract: The Visceral Leishmaniasis (VL) is a anthropozoonosis in Brazil that is transmitted by females of Lutzomyia longipalpis, the occurrence of which depends on the presence of species involved in the transmission chain and favorable environmental conditions. In the city of Mossoro, in Northeast Brazil, VL is endemic, its vector is presented in significant numbers throughout the year and has its development tied to wetlands, and their increased density in post-rain periods.

These factors justify the present study, which was developed in areas near the river that passing through the city, as are wetlands and suffering flooding in times of heavy rains. Based on the above, this study aims to know the level of knowledge of Community Health Agents who work in these areas and are responsible for the activities of disease prevention and health promotion, including VL. To that end, we chose the three city districts that are subject to these floods and the study was conducted with 12 Community Health Agents who agreed to answer an interview with questions covering aspects of knowledge about the transmission of VL. Of all respondents said they had heard about VL (100%), but only 8 (66.6%) reported that the man and the dog get sick and 2 (16.6%) reported being the man and other animals, but did not specify which animals.

When asked if they knew how the disease was transmitted answered all know (100%), of these, 11 (91.6%) said it was caused by a mosquito bite, but not be cited the sandfly. When asked if they knew where the sandfly females put their eggs, 7 (58.3%) said they knew the answer, but only 4 (33.3%) responded that this stance is on organic matter, of which 1 (25.0%) exemplified leftover fruit and 1 (25.0%) cited feces and uneaten food. When asked about the activity time vector only 4 (33.3%) responded evening. And on ways to reduce the presence of the vector in the environment, 10 (83.3%) responded that cleaning the environment is important. This result demonstrates the lack of professionals working with public health, on issues involving the disease, this fact implies the permanence of VL emphasizing that this is a city endemic for the

disease. It can be seen then, the importance of training policies, seeking to spread the knowledge about how to prevent the formation of environments conducive to the development of the vector.

- CONTROL PROGRAMS [1113] P 875 - THE SURVEILLANCE AND CONTROL OF VISCERAL LEISHMANIASIS IN ENDEMIC AREAS OF THE CITY OF MOSSORO, RIO GRANDE DO NORTE, BRAZIL COSTA, K.F.L.1; AMORIM, C.F.1; SILVA, L.F.1; KAZIMOTO, T.A.1; REGINALDO, M.L.1; ALBANO AMORA, S.S.1; DE SOUZA, C.S.F.2; NOGUEIRA FREITAS, Y.B.1; BARBOSA, S.R.1; DE MACEDO, L.B.1; FERNANDES, A.M.M.M.3; ALVES, N.D.1; CARNEIRO FEIJÓ, F.M.1; MORAIS, M.R.P.T.1 1.UNIVERSIDADE FEDERAL RURAL DO SEMI-ÁRIDO - UFERSA, MOSSORÓ, RN, BRAZIL; 2.DEPARTAMANETO DE PROTOZOOLOGIA - FUNDAÇÃO OSWALDO CRUZ - FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.SECRETARIA DE VIGILÂNCIA A SAÚDE - PREFEITURA MUNICIPAL DE MOSSORÓ, MOSSORÓ, RN, BRAZIL.

Keyword:prenvention; community health agents; calazar

Abstract: The Visceral Leishmaniasis (VL), a disease chronic and lethal commonly caused by the protozoan Leishmania chagasi and transmitted through the bite of sandflies insects, has occurrence associated with several factors, like the discontinuity of control actions related to the disease relevant. In this sense, this research included the completion of semi-structured interviews with professionals working with surveillance, control and health promotion of Basic Health Units in endemic districts for VL of the city of Mossoro in northeastern Brazil. Two groups of professionals were included in the study, Agents Endemic (AE) and Community Health Agents (CHA), totaling 38 professionals, who underwent an interview addressing questions about the activities of Surveillance and Control of the VL, and training courses promoted by the municipality about VL. After the interviews, it was revealed that 15.8% (6/38) of the agents not received training to work with the population in preventing the disease. Among the professionals who received training, they were exclusively in the form of courses, 71.9% (23/32) claimed that these activities included information about the human and canine VL. However, 53.1% (17/32) mentioned that these courses have rare frequency, over 4 years between them. Even among trained professionals, 71.9% (23/32) believe they can absorb the information passed on during training activities, which suggests that if these occur regularly, would bring good results to the work of the agents in the prevention and control of VL. Moreover, 34.2% (13/38) and the ACS stated that AE is not actively search for cases of human and canine LV, and the same percentage of respondents believe that search is only performed when there is suspicion. About the practice of Health Education for VL, 84.2% (32/38) confirmed its realization, however, 47.4% (18/38) said they did not perform any type of surveillance activity for canine VL and 84.2% (32/38) also did not perform for human VL. These responses contrasted with the fact that 47.4% (18/38) of agents have confirmed cases of human VL and / or dogs in the area in which they work, proving the importance of conducting the Surveillance and Control of the disease. Thus, one realizes the need for a greater incentive for the municipality that the CHA and AE work to reduce or prevent the expansion cycle of VL in the city of Mossoro, so that these agents also need to perform activities more emphatic and comprehensive.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1117] P 876 - RESIDUAL EFFECT AVALIATION OF BENDIOCARB, CARBAMATE INSECTICIDE GROUP ABOUT FATALITY RATE OF SANDFLY VECTOR OF TEGUMENTARY LEISHMANIASIS IN CEARÁ STATE, BRAZIL. CARANHA, L.; CUNHA, J.L.; RODRIGUES, L.O.; CUNHA, G.D.; LIMA, J.W.O.; DE ARAÚJO, J.M.P.; DE MOURA, F.K.; C. MOREIRA, P.A.; G. DE SOUSA, F.D. SECRETARIA DA SAÚDE DO CEARÁ, FORTALEZA, CE, BRAZIL; ; ; ; ; ; ; ; .

Keyword:sandfly; control; tegumentary leishmaniasis

Abstract: Tegumentary Leishmaniasis is endemic in Ceará State. It has a large distribution registering 2917 cases from 2010 to 2012. It has showed outbreaks in some municipalities with incidence rate concerning 39,25 cases in 100.000 inhabiting in the last three years (SESA-CE). In Brazil chemical control using residual action insective is the action recommended in vector control in coletive protection way. This study has na objective to avoid or reduce the contact between transmitter vector and human population and therefore diminishing the disease transmission risk. This present study objectives to study residual effect of Bendiocarb (Bayer) PM 80% insective in 300mg ia/m2 dose through biology proofs in wall in na Amarican Tegumentary Leishmaniasis (ATL) endemic area using Bendiocarb. In tests were used males and females insects from natural population always captured in the same place. It was done four tests Each one with ten samples with 20 or 30 specimens in each one. Five samples were exposed inside home, which three in walls having different heights of 0,50, 1,0 and 1,5 m and one in a wood surface and one control. Likewise were exposed five remander ones around home.Test was done in Buira Grande place in Viçosa do Ceará municipality. Insects were exposed during an hour and final report was done twenty four hours. The tests showed that fatality rates were good sooner after spraying (24 h) from 88,9 to 100 in both inhabitings and surfaces. In thirty two days fatality varied from 96,2 to 100 % in wall inside home and 100% in wood surface. After sixty days it was from 13 to 89,3% in wall and 94,9% in wood surface. In the last test 95 days after spraying the fatality inside home reached from 19,2 to 30% fatality in wall and 100% in wood surface. Around home fatality was 14,7 to 57,7% in wall and 94,4% in wood surface. Fatality in controls was from 2,9 to 10% in all tests. Samples were composed by 43,32 Lutzomyia whitmani, 55,81 L. migonei and 0,87 L. longipalpis. The low fatality observed in wall surface in the last test direct us to do cicles until 60 days after spraying. There is also a necessity of complex studies in order to verify how a vector population recover in treated areas. Those knoledges will optimizing cost and benefit in sandfly control.

- EPIDEMIOLOGY [1121] P 877 - CONCLUSIONS ON THE EPIDEMIOLOGY AND POPULATION GENETICS OF TEGUMENTARY LEISHMANIASIS BASED ON MICROSATELLITE ANALYSIS – A FIRST REPORT FROM PARAGUAY ODDONE, R.1; ANDRÉS, C.2; SCHÖNIAN, G.3; KUHLS, K.4 1.INSTITUTO DE INVESTIGACIONES EN CIENCIAS DE LA SALUD, UNIVERSIDAD NACIONAL DE ASUNCIÓN, IICS, ASUNCIÓN, PARAGUAY; 2.LEISHMANIASIS CONTROL NATIONAL PROGRAMME, SENEPA, MINISTRY OF PUBLIC HEALTH, ASUNCIÓN, PARAGUAY; 3.INSTITUTE OF MICROBIOLOGY AND HYGIENE, CHARITÉ UNIVERSITY MEDICINE, BERLIN, GERMANY; 4.TECHNICAL UNIVERSITY OF APPLIED SCIENCES, MOLECULAR BIOTECHNOLOGY AND FUNCTIONAL GENOMICS, WILDAU, GERMANY.

Keyword:microsatellite analysis; population genetics; paraguay

Abstract: In Paraguay tegumentary leishmaniasis is endemic in many areas of the Oriental Region (San Pedro, Canindeyú, Caaguazú, Alto Paraná). The number of registered cases during the last decade ranged between 1251 and 231 cases per year, with an average of 24.5 % of mucocutaneous cases, which is far above all the other countries of Latin America. All cases were caused by Leishmania (V.) braziliensis. So far, information about epidemiology of leishmaniasis and the genetic variability of the parasites in Paraguay is scarce. In this first population genetic study, 46 strains of L. (V.) braziliensis from nine departments, including 17 strains of a big outbreak in 2000, and nine strains from neighbouring endemic regions of Bolivia, Argentina and Brazil were analysed by using multilocus microsatellite typing (MLMT).

This study was aimed to assess the genetic variability and population structures of L. (V.) braziliensis in Paraguay and to correlate them with the geographical origins and with different clinical manifestations, such as development of mucocutaneous lesions, and also to compare Paraguayan L. (V.) braziliensis with strains from neighbour regions in Brazil, Bolivia and Argentina in order to identify the origin of outbreaks and to track the mode of spread of the genotypes MLMT data, obtained by PCR and fragment analysis, were analysed by using a Bayesian model-based clustering algorithm, genetic distance analysis and factorial correspondence analysis. According to all three analysis approaches strains split into two main populations A (20 strains from Paraguay, 2 from Argentina, 1 from Bolivia, 3 from Brazil) and B (26 strains from Paraguay, 2 from Bolivia, 1 from Brazil) that correlate with the geographical origin of the samples. All but one strains obtained from the centre of the 2000 outbreak belong to population A. Further analysis revealed three sub-populations in each of the populations A and B. There was no correlation between population assignment and clinical picture, suggesting that other factors might play key roles in the determination of the clinical outcome. It has to be established whether the two main populations are determined also by other epidemiological factors, as e.g. transmission by different vectors. This work showed a considerable amount of genetic variability in Paraguayan L. (V.) braziliensis, similarity of Paraguayan genotypes to those circulating in endemic regions of neighbouring countries and the usefulness of MLMT in monitoring epidemic outbreaks.

- OTHER SPECIAL TOPICS [1130] P 878 - KNOWLEDGE AGENTS ON ENDEMIC TRANSMISSION OF VISCERAL LEISHMANIASIS IN BRAZILIAN CITY FOR ENDEMIC DISEASE AMORIM, C.F.1; KAZIMOTO, T.A.1; DE LIMA COSTA, K.F.1; SILVA, L.F.1; REGINALDO DE SOUSA, M.L.1; ALBANO AMÓRA, S.S.1; NOGUEIRA FREITAS, Y.B.1; BARBOSA, S.R.1; DE MACEDO, L.B.1; ALVES, N.D.1; CARNEIRO FEIJÓ, F.M.1; FERNANDES, A.M.M.M.2 1.UNIVERSIDADE FEDERAL RURAL DO SEMI-ÁRIDO – UFERSA, MOSSORÓ, RN, BRAZIL; 2.SECRETARIA DE VIGILÂNCIA À SAÚDE – PREFEITURA MUNICIPAL DE MOSSORÓ/RN, MOSSORÓ, RN, BRAZIL.

Keyword:health professional; sandfly; environment

Abstract: The Visceral Leishmaniasis (VL) is aanthropozoonosis and the etiological agent in Brazil, is the protozoan Leishmania infantum chagasi that is mainly transmitted through the bite of female phlebotomine hematophagous species Lutzomyia longipalpis. In the state of Rio Grande do Norte, Northeastern Brazil, VL came to represent an important public health problem in 1989, since then the disease is considered endemic in this state, especially for the city of Mossoro.

Based on the above, this study aims to know the level of knowledge of agents who work in the city Endemics Mossoró, these professionals are responsible for the activities of prevention and control of endemic diseases common in each region. In all the Mossoró for 20 agents who work directly with Endemics VL and of those, eight agreed to participate in the interview that addressed aspects of the transmission of VL. All respondents reported they had heard about the disease. When asked about who might get sick of VL, 2 (25.0%) responded that humans and other animals, without specifying how and 6 (75.0%) answered the man and the dog, one of those said the fox and the cat another. All respondents know that the disease is transmitted by the bite of a mosquito that transmits specifically the VL, but not be cited the sandfly. However, all respondents also have seen the vector, giving accurate information about their morphology.

When asked about where the females lay their eggs in sand fly and knew the environments favorable to the emergence of vector, 7 (87.5%) answered correctly. When asked about the

hours of activity vector, 5 (62.5%) reported being nocturnal. With regard to what to do to decrease the presence of the vector environments, 6 (75.0%) answered the realization cleanliness of the premises and of these, 4 (50.0%) also cited the completion of fumigation environments. Therefore, it is clear that the agents have extensive knowledge about Endemics VL, but still need more training to be able to instruct and work with the population of the areas that are included, more appropriately and consistently.

- OTHER SPECIAL TOPICS [1133] P 879 - KNOWLEDGE OF COMMUNITY HEALTH WORKERS ON VISCERAL LEISHMANIASIS IN ENDEMIC AREAS OF THE CITY OF MOSSORO, RIO GRANDE DO NORTE, BRAZIL AMORIM, C.F.1; SILVA, L.F.1; DE LIMA COSTA, K.F.1; KAZIMOTO, T.A.1; REGINALDO DE SOUSA, M.L.1; ALBANO AMÓRA, S.S.1; NOGUEIRA FREITAS, Y.B.1; BARBOSA, S.R.1; RIBEIRO, T.T.1; ALVES, N.D.1; CARNEIRO FEIJÓ, F.M.1; FERNANDES, A.M.M.M.2 1.UNIVERSIDADE FEDERAL RURAL DO SEMI-ÁRIDO – UFERSA, MOSSORÓ, RN, BRAZIL; 2.SECRETARIA DE VIGILÂNCIA À SAÚDE – PREFEITURA MUNICIPAL DE MOSSORÓ/RN, MOSSORÓ, RN, BRAZIL.

Keyword:health care professionals; health education; calazar

Abstract: Surveillance and control of Visceral Leishmaniasis (VL) involve the articulation of various professionals and institutions in promoting health education as a way of disseminating information about the disease and environmental conditions relevant to reduce the risk of emergence of vector. Aiming to assess the knowledge of health professionals about the VL, a semistructured interview was conducted with the Community Health Agents of the Basic Health Units for VL endemic districts of the city of Mossoro, Rio Grande do Norte, Brazil totaling 30 professionals. The interview covered questions about knowledge of VL, as transmission, symptoms, diagnosis, treatment, and prevention and control. After the interviews, it was found that 96.7% (29/30) of the Community Health Agents knows zoonotic disease; 63.3% (19/30) know some symptoms of disease in humans, whereas 10% (3/30) not aware of any symptoms.

About diagnosis, 66.7% (20/30) knew only that this is achieved through blood test and 13.3% (4/30) did not know how to get. Most respondents, 83.3% (25/30), stated that the disease is curable and 50.0% (15/15) mentioned that the treatment is carried out based medicines such as antibiotics, showing lack of knowledge of these professionals regarding the treatment of this important zoonosis. Regarding prevention, 63.3% (19/30) of Community Health Agents know there is no vaccine to prevent the occurrence of VL. About the transmission, 16.7% (5/30) of agents said that the disease is transmitted by direct contact with infected dogs, which consists of an index worrisome, since a wrong information about the mode of transmission may cause loss or failures in preventing and controlling the disease. Moreover, 30.0% (9/30) of Community Health Agents stated not knowing cite any measures to prevent the occurrence of disease. Thus, we can conclude that it is still a need for greater dissemination of disease to health professionals working in the surveillance and control of disease and/or health promotion, since these professionals represent one of the most responsible for bringing the information to the population and that disease control directly depends on the knowledge of these actors in the matters relating to the disease.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1140] P 880 - AVALIATION OF INSECTICIDE APPLICATION IN VECTOR SANDFLIES POPULATION CONTROL OF TEGUMENTARY LEISHMANIASIS IN CEARÁ STATE, BRAZIL

RODRIGUES, L.O.; SOUSA, L.C.; CUNHA, J.C.; DIAS, G.V.; MOREIRA, P.A.; DE SOUSA, F.D.; MOURA, F.K.; ARAÚJO, J.M.; LIMA, J.W.O. SECRETARIA ESTADUAL DE SAÚDE DO CEARÁ, FORTALEZA, CE, BRAZIL; ; ; ; ; ; ; ; .

Keyword:sandflies; avaliation; tegumentary leishmaniasis

Abstract: American Tegumentary Leishmaniasis in Ceará State showed in the last five years an average of 1003,3 cases with an incidence rate concerning 67.53 in 100.000 inhabiting. Chemical control is guided by Health Ministry (HM) since it is not intended in wild areas and that decisions must be taken from joint analysis of epidemiological and entomology datum. This study objectived to evaluate in an entomology way the treatment using Carbanates insecticide – Bendiocarb (Bayer) PM 80% in 300mgia/m² dose through sistematic collections of sandflies in an ATL endemic area treated with the same insecticide. In withdrawals were used light trap called CDC capturing in 12 hours begining from 6:00 pm and kept at 6:00 am in the following day. It was done four registrations. One registration one week before treatment and the others in three consecutive nights in four places in Viçosa do Ceará municipality. The places are: Buirinha (control area), Buira Grande, Porteiras and Canto da Buira. In each place was chosen a house that could show propitious caracteristics to sign the sandflies presence. In the registration that revealed treatment were collected 1520 sandflies distributed in a such way: Buira Grande (264 specimens), Porteiras (39) and Canto da Buira (31 sandflies). In Buirinha control area were collected 1186 specimens. No one registration verified rainings. In 24 hours registration after treatment we could observe a 81,64% fall with only 61 specimens collected in treated areas. Therefore in the third registration 30 days after we could see this number came back to the same begining status added to 5,4% (352 specimens) in sandflies collected, returning to stabilize and maintaining this pattern until the last registration , which was 60 days after with 305 collected specimens. During avaliation it was collected 5708 sandflies belonged to 6 species, highlighted the predominance of Lutzomyia whitmani (58,29%) and L. migonei (38,75%) collected inside and arround home in all avaliations. In theses conditions we can conclude in case of using chemical control in a right time, transmission will suffer a great impact that can be broken. However, even insecticide has a residual effect it will not ensure the controlof populations that recover in short period of time.

- OTHER SPECIAL TOPICS [1142] P 881 - VISCERAL LEISHMANIASIS: 100 YEARS FROM THE FIRST REPORT IN THE AMERICAS ODDONE, R. INSTITUTO DE INVESTIGACIONES EN CIENCIAS DE LA SALUD, ASUNCION, PARAGUAY.

Keyword:visceral leishmaniasis; migone; paraguay

Abstract: In the history of leishmaniasis, the year 1913 has led to the disclosure of a breakthrough: the report of the first human case of visceral leishmaniasis (HVL) diagnosed in the Americas, whose merit was awarded to the Paraguayan Dr. Luis Enrique Migone, who published it in the Bulletin de la Société de Pathologie Exotique 6: 118-120. The purpose of this review was to recall the fact, as well as to discuss ways to control this neglected disease. Placing us at the historic moment, it was considered an act of courage and bravery make discovering and publishing the finding made in 1911 of a disease hitherto known only in Asia. Nor was conducive the politicalmilitary environment rather tense of Paraguay from 1911 to 1912. The first cases found in Brazil were described over 20 years later.

The case diagnosed by Migone was an Italian immigrant living in Brazil, who came from Corumbá (Mato Grosso do Sul) with the characteristic symptoms of HVL. Migone observed Leishmania in blood smears, liver puncture and spleen puncture, which confirmed the diagnosis. Treated with arsphenamine and despite showing improvement, the patient died a month later from an acute diarrhea.

Considering that LVH is a disease with high mortality rate, and paradoxically very neglected in all countries, this is an important opportunity to highlight the true approach that deserves the population against this public health problem. Control campaigns are not reducible to attend only the human component, neglecting the vector component and domestic reservoir. Thus, studies have been published (reviewed by Romero and M. Boelaert in PLoS Negl Trop Dis 4 (1), 2010) showing the relative effectiveness of integrated interventions, ie addressing all components of the transmission, in an area wide. These references indicate that combined interventions (against vectors and reservoirs) have positive results in reducing the incidence of new infections. This means that working hard as a team, all technicians, health professionals and researchers can gain control of HVL if working seamlessly. Our people deserve this effort.

- EPIDEMIOLOGY [1145] P 882 - TITLE: CUTANEOUS LEISHMANIASIS IN CALAKMUL CAMPECHE, MÉXICO OVER A PERIOD OF 2 YEARS (2006–2008) MONROY OSTRIA, A.M.1; HERNANDEZ RIVERA, M.P.2; HERRNANDEZ MONTES, ,.O.2; CHIÑAZ PEREZ, A.3; SANCHEZ TEJEDA, G.4; WONG RAMÍREZ, C.2; MARTINEZ, M.A.2 1.ENCB, INSTIUTO POLITÉCNICO NACIONAL, MEXICO, MEXICO; 2.ENCB, INSTITUTO POLITECNICO NACIONAL, MEXICO, MEXICO; 3.AYUNTAMIENTO DE CALAKMUL, SECRETARIA DE SALUD, CAMPECHE, MEXICO; 4.SECRETARÍA DE SALUD, MEXICO, MEXICO.

Keyword:cutaneous lieshmaniasis; epidemiology. calakmul; campeche, mexico

Abstract: In the sylvatic region of the Yucatan peninsula of Mexico, an endemic focus of cutaneous leishmaniasis (CL), the Calakmul district of Campeche State, is affected by the localized cutaneous and mucocutaneous clinical forms of leishmaniasis. In the present study, One hundred and forty-six individuals with skin lesions suggestive of Leishmania infection were evaluated over a period of 2 years (2006–2008).

Aspirates taken from the cutaneous lesions were inoculated in Senekjie’s medium and golden hamsters. The culture detected 42% of the positive samples and 92% of the inoculated hamsters were positive. DNA from the aspirates amplified by PCR detected 88% of the infected patients with L. (L.) mexicana, 25% infected with Mexican strains of L. (L.) mexicana, and 12% with members of the L. braziliensis complex; 7% of the infected patients had a mixed infection with L. mexicana and L. braziliensis complex. The diagnose of CL of 57 / 146 was proximately 40% of CL infection rate. The mainly risk factors found are human colonization of large areas of previously untouched forests spending periods in the forest for cultivation of agricultural crops, increase of military troop maneuvers.

After treatment with meglumine antimoniate (Glucantime), 95% of the patients with confirmed CL was healed.

- EPIDEMIOLOGY [1146] P 883 - EPIDEMIOLOGICAL STUDY BY REGION OF BRAZIL ON VISCERAL LEISHMANIASIS VIEIRA, A.T.P.; DOS SANTOS, F.R.B.; BASTOS, C.E.; NEGROMONTE, G.R.P.; MENDONCA, M.S. UNIVERSIDADE FEDERAL DA PARAIBA, JOAO PESSOA, PB, BRAZIL.

Keyword:visceral leishmaniasis; epidemiology; disease

Abstract: Introduction Visceral leishmaniasis (VL) is a typically rural disease, distributed mainly in tropical and subtropical regions. Brazil represents 97% of all cases of VL in the Americas. With the urbanization, the VL suffered an epidemiological transition and has reached urban areas, where there are people sanitary, social and economically excluded. The epidemiology is important, since it provides a higher subsidy for the presumptive diagnosis and serves as guidance for health politics.

Objectives This paper aims to demonstrate the epidemiological profile of VL stratified by Brazilian regions, from 2007 to 2011.

Development This is a descriptive study with a qualitative and deliberative approach. Cases not resident in Brazil and duplicate information have been removed. It has been made graphics to analyze the results and understand its importance.

Results Leishmaniasis prevailed among those with incomplete primary education, among males, with the percentage of 72,7% in the South and 64,3% in the Midwest. It has prevailed also among mixed races. In this case, the percentage was of 74,86% in the Northeast and 80,36% in the North. Only in the South, the percentage was higher for the white race, with 78,12%. Regarding the number of pregnant women, there was 58 cases in the last 5 years, in which was registered only one case in the South.

Analyzing the age groups, the case between children until a year old have prevailed, especially in the North and Northeast. The northeast has shown the higher absolut number in co-infection with HIV, with 491 cases, whereas the South has shown the higher relative number, with 15,62%. Most of the cases have been ignored or in blank, what makes the study more difficult.

Itâ&#x20AC;&#x2122;s shown that the parasitic diagnosis has not been done in more than 43% of the cases, mostly in the South where the number achieved 57,5% of all cases without this diagnosis. When the test was done, the positive result prevailed. 87,24% of all cases have been confirmed.

The situations in which new cases occurred prevailed to the recidivists and transferred cases.

Most cases were healed and the dropout rate was below 1% in most regions observed, highlighting the lowest dropout rate in the Northeast, with 0,49%.

Conclusion VL is now distributed by the urban peripheries and the study of epidemiology is important for observing the extent and spread of VL in the country, thus guiding the actions combat the disease - CONTROL PROGRAMS [1164] P 884 - PREVENTING ZOONOTIC CANINE LEISHMANIASIS IN NORTHEASTERN BRAZIL: PET ATTACHMENT AND ADOPTION OF COMMUNITY LEISHMANIA PREVENTION ESCH, K.1; PONTES, N.2; ARRUDA, P.1; OCONNOR, A.1; MORAIS, L.2; JERONIMO, S.2; PETERSEN, C.1 1.IOWA STATE UNIVERSITY, AMES, UNITED STATES; 2.UFRN, NATAL, RN, BRAZIL.

Keyword:canine; topical permethrin; treatment

Abstract: Visceral leishmaniasis (VL), caused by Leishmania infantum chagasi (L.i. chagasi syn. infantum) in Northeastern Brazil, was responsible for 51,000 new VL cases from 1980--2003.

Household presence of L. infantum-infected dogs is a major risk factor for human infection.

Despite culling of dogs based on seropositivity, canine L. infantum seroprevalence remains near 20%, suggesting dog culling is ineffective for preventing VL spread. We administered a crosssectional survey to 224 households within 300 meters of VL human patient homes diagnosed within the last year. The goal was to develop a model for voluntary preventative utilization based on characteristics and motivations of dog owners. We identified that owner knowledge deficiencies regarding canine transmission of L. infantum associated with increased risk of dog infection (OR=3.681, CI=1.223, 11.08). Higher owner education was associated with decreased levels of dog seropositivity (OR=0.40, CI=0.20, 0.81). Pet attachment (p=0.036) and perception of risk/disease knowledge (p=0.040) were significantly associated with willingness to voluntarily purchase canine VL prevention. These results highlight the importance of owner attachment to their pet in implementing reservoir-targeted zoonotic visceral leishmaniasis prevention.

- EPIDEMIOLOGY [1177] P 885 - VISCERAL LEISHMANIASIS: EPIDEMIOLOGY OF A NEGLECTED DISEASE IN CHILDREN, ALAGOAS, NORTHEASTERN BRAZIL, FROM 2007 TO 2012.

FREITAS, I.S.1; GOES, J.J.1; ALVES, W.A.1; PEIXOTO, V.S.1; FERREIRA, I.G.1; SOUZA, C.M.1; CORTEZ, A.M.C.1; SOBRINHO, A.C.R.1; MACEDO, D.S.1; SÁ, W.T.1; GOMES, A.R.F.1; CALHEIROS, M.B.1; LESSA, M.M.1; SILVA, M.E.2 1.UFAL, MACEIÓ, AL, BRAZIL; 2.NUSP, MACEIÓ, AL, BRAZIL.

Keyword:visceral leishmaniasis; children; epidemiology

Abstract: Visceral leishmaniasis (VL) is an anthropozoonosis with broad geographic distribution worldwide. In Latin America there is a predominance of cases in Brazil (90% cases) where it is an obligatory notifiable disease. Recent data released by Brazil’s Ministry of Health (MS)

indicates the northeast, composed of 11 federal units (UF), representing approximately 50% of notifications, with an average mortality of 6% / year. The dog (Canis lupus familiaris) is the main domestic reservoir of Leishmania (L.) infantum chagasi (Lainson & Range, 2005). MS defines LV: clinically - "person from area where occurs transmission with fever and splenomegaly"; laboratory - person with compatible clinical and immunological or parasitological positive diagnostic. With the lowest UF Human Development Index (HDI) of the country, Alagoas (AL) has characteristics that suggest a limited panorama of the control and prevention of disease, especially in children.

This study describes clinical and epidemiological aspects of VL in children 10 years, reported in AL, from 2007 to 2012.A descriptive, cross-sectional study was conducted with data from the Information System for Notifiable Diseases MS - SINAN. Inclusion criteria: records with clinical or laboratory confirmation. Variables: area of residence, sex, age, clinical features (available in SINAN), drug use and clinical evolution. During the study period, 265 cases were reported by LV, and 111 (53%) in children 10 years. 79% of them lived in rural areas. 56% occurred in boys. The mean age for boys was between 4 years and 3.5 years in girls (p = 0.49).

Frequency of clinical manifestations: fever (99%), pallor (96%), hepatosplenomegaly (94%), weight loss (93%) and weakness (76%). Drug of first choice: pentavalent antimony (85%), amphotericin B (10%), untreated (1.4%). For clinical evolution, only 62 (56%) had data records, as follows: 33 (52%) cures, 1 (2%) abandonment, 24 (38.7%) others. Lethality of 6.5% (4) is noteworthy. AL still features an epidemiological standard where children represent the majority of cases with severe clinical features, high mortality and socioeconomic scenario favorable to increased incidence and lethality LV in the coming years. The study points to greater integration between primary care and health surveillance to strengthen health care for early recognition and treatment of VL child.

- EPIDEMIOLOGY [1179] P 886 - LEISHMANIASIS’ MORTALITY: ANALYSIS OF THE INFORMATION SYSTEM FOR DEATH. PEIXOTO, V.S.1; LESSA, M.M.1; GOMES, A.R.F.1; CALHEIROS, M.B.1; GÓES, J.J.1; FREITAS, I.S.1; FERREIRA, I.V.G.1; SOUZA, C.M.1; CORTEZ, A.M.C.1; SOBRINHO, A.C.R.1; MACEDO, D.S.1; DE SÁ, W.T.1; SILVA, M.E.B.2; ALVES, W.A.1 1.UFAL - FACULDADE DE MEDICINA, MACEIO, AL, BRAZIL; 2.NÚCLEO DE SAÚDE PÚBLICA, MACEIO, AL, BRAZIL.

Keyword:leishmaniasis; epidemiology; mortality

Abstract: Leishmaniasis (Leish) is a zoonose with broad geographic distribution in Brazil, representing serious public health problem in human and veterinary medicine. It is known that Leish have universal susceptibility and the infection does not confer immunity, however visceral leishmaniasis (VL), if untreated, can progress to death. In cutaneous leishmaniasis (CL), monitoring the clinical course is essential to reduce the severe form (mucosal), prevent deformities and death. In Brazil, inputs for laboratory diagnosis (serological and parasitological) and drugs for treatment are provided free by the public health system through primary and hospital care. However, there is an important magnitude in registration of deaths at the Department of Informatics MS - Datasus. The study aimed to characterize Leish’s mortality in Alagoas, using data from the Mortality Information System (SIM), from 2001 to 2010. This is a descriptive transversal study. 52 death records have been verified whose underlying or final/immediate cause (disease that initiated the chain of morbid events leading directly to death) has received one of the ICD10 (International Classification of Diseases) for Leish (B550: LV; B551: Cutaneous Leish; B559: unspecified Leish). 42 (81%) were B550, 5 (9.5%) B551, 5 (9.5%) B559. LV: 55% men, mean age = 22 years (median = 9 years) (range = 0.7 to 73 years),

45% women, mean age = 14 years (median = 6 years) (range = 0.6 - 6 years). Education: only 14 (33%) with data records. Race: only 28 (67%) records with data, predominance of black and brown races (78%). Place where death occurred: 40 (95%) in the hospital. LT: Year of death: 2002 (1) 2001 (1) 2006 (1) 2007 (2); 3 (60%) men; Race: mulatto 75%; Education: 60% illiterate; Age: range: 24 -84 years, median = 48 years. The analysis of information from Health Information Systems is critical to the development of public policies and to better understand the relationship between causal determinants and morbimortality in a population. Secondary data, systematized and analyzed from SIM can contribute greatly to better understand Leish’s mortality in Alagoas, especially LT whose prognosis should be cure. However, it is essential that the quality of data from death certificates is ensured, once it’s source for SIM. - RESERVOIRS [1180] P 887 - LEISHMANIA INFANTUM AND DIROFILARIA IMMITIS INFECTION IN DOMESTIC AND STRAY CATS FROM SOUTHERN PORTUGAL MAIA, C.1; RAMOS, C.2; COIMBRA, M.3; SOUSA, B.4; CRISTÓVÃO, J.M.5; CORTES, S.5; MARTINS, A.2; CAMPINO, L.6 1.UEI PARASITOL MÉDICA/CMDT/INSTITUTO HIGIENE MEDICINA TROPICAL-IHMT/ UNIV NOVA LISBOA-UNL. FMV/ULHT, LISBOA, PORTUGAL; 2.FMV, UNIVERSIDADE TÉCNICA DE LISBOA, LISBOA, PORTUGAL; 3.CLÍNICA VETERINÁRIA PORTO SEGURO, OLHÃO, PORTUGAL; 4.CMDT/FACULDADE DE PSICOLOGIA E CIÊNCIAS DE EDUCAÇÃO, UNIVERSIDADE DE COIMBRA, COIMBRA, PORTUGAL; 5.UEI PARASITOL MÉDICA/CMDT/INSTITUTO HIGIENE MEDICINA TROPICAL-IHMT/ UNIV NOVA LISBOA-UNL, LISBOA, PORTUGAL; 6.UEI PARASITOLOGIA MÉDICA/IHMT/UNL. DEP CIÊNCIAS BIOMED E MED/UNIVERSIDADE DO ALGARVE, LISBOA, PORTUGAL.

Keyword:leishmania infantum; dirofilaria immitis; cat

Abstract: Human visceral and cutaneous leishmaniases are endemic in Portugal. Leishmaniasis and heartworm are vector-borne parasitic diseases caused by a protozoan, Leishmania infantum, and by a nematode, Dirofilaria immitis, respectively. Despite the fact that these emerging diseases are well known in the canine population, they are often underestimated in the feline population, likely due to the unknown of clinicians and especially the owners. The aim of this study was to determine the prevalence of L. infantum and D. immitis infection in the feline population from southern Portugal. From 2011 to 2012, 217 cats from Olhão were analysed. Infection with L. infantum was analysed by direct agglutination test-DAT (cut-off 1:100) and by PCR-RFLP for genotyping. D. immitis infection was determined using the SNAP ® - Feline Triple®. The Chisquare test was chosen in order to explore possible associations among molecular and serological findings and the variables in the study. Fisher's Exact Test was used when the conditions for the realization of the first were not present. The prevalence of Leishmania sp. infection obtained in this study was 5%. Specific anti-Leishmania antibodies were detected in 9 (4%) of the 215 cat´sera. The analysis of the enzymatic restriction of the PCR product showed that parasites belonged to genotype A, the most frequent in Portugal among Leishmania strains isolated from humans, dogs and sand flies. The seroprevalence for D. immitis infection was 6%.

Statistical analysis found a correlation between heartworm and feline leukemia virus (p = 0.049) while no correlation between Leishmania and the other studied variables were observed. As far as we are aware this is the first time that L. infantum and D. immitis infections were detected in the feline population living in southern Portugal. The existence of positive cases emphasizes the need to alert both veterinary community and owners for the risk of infection in the feline population by these two parasites, and consequently, there is a need to implement prophylactic measures to protect animals and people. Nevertheless, our data reinforce the idea that both parasitosis should be included systematically in clinical practice as differential diagnoses, especially in areas where they are endemic.

Acknowledgements: To all cat´s owners and veterinarians that participated. This work was supported by CMDT and IDEXX Laboratories. C. Maia (SFRH/BPD/44082/2008) holds a

fellowship from Fundação para a Ciência e a Tecnologia, Ministério da Ciência, Tecnologia e Ensino Superior, Portugal.

- EPIDEMIOLOGY [1185] P 888 - EPIDEMIOLOGICAL CHARACTERIZATION OF DEATHS FROM VISCERAL LEISHMANIASIS, ALAGOAS, NORTHEAST, BRAZIL, FROM 2001 TO 2012 SOBRINHO, A.C.R.; MACEDO, D.S.; ALVES, W.A.; GOES, J.J.; LESSA, M.M.; CALHEIROS, M.B.; FREITAS, I.S.; FERREIRA, I.V.G.; SOUZA, C.M.; SÁ, W.T.; GOMES, A.R.F.; PEIXOTO, V.S.; SILVA, M.E.B.; PEREIRA, C.W. UNIVERSIDADE FEDERAL DE ALAGOAS, MACEIÓ, AL, BRAZIL.

Keyword:epidemiology; visceral leishmaniasis; mortality

Abstract: Visceral Leishmaniasis (VL) is a chronic and systemic zoonosis that has a high mortality rate (90%) when untreated. In Brazil, the disease is presenting a large expansion to urban areas that classifies it as a serious public health problem. This study aimed to characterize, clinically and demographically, deaths from LV in Alagoas, registered at the Disease Notification System (SINAN) of the Ministry of Health (MS) from 2001 to 2012. We conducted a descriptive, transversal study. 1168 cases of VL, and 40 subsequently died (mortality: 3.5%) were recorded.

Gender: 73% male. Death age: mean = 19, median = 13 years. Average age for males = 21 years (median 17 years) and average age for female = 14 years (median = 2 years) (p = 0.33). Age range: 1 year: 7%; 1-10 years: 37%, 11-30 years: 27%, 31-50 years: 21%, 50 years: 8%.

Race: 88% browns, 2% black, 2% white, 8% ignored. Education: 17% illiterate, 30% 1-7 years of study. Area of residence: 63% rural, 32% urban and peri 5%. Treatment: 50% started with pentavalent antimony, 22% with amphotericin B, pentamidine with 2%, 8% were ignored. It’s important to note that 18% did not use medication. On failure of initial treatment, 12% used amphotericin B, 5% used another drug. It is noteworthy that 25% did not use other medication, and 58% of records had no data on second-line medication. The period (days) between notification and death was: mean = 19 days, median = 9 days, maximum = 167 days. 87% of patients died within the first fifty days between registration on SINAN and the actual death. It is relevant to note that 5% took more than one hundred days to die. Between 2007 and 2012, 25% started treatment within two days after notification, 31% between the third and fifth day, 19% between the sixth and tenth day, the rest was ignored. The profile of deaths is similar to what was described in the national literature from Northeast, focusing on children, male, living in rural areas. The pentavalent antimony was the drug of first choice and the time between notification and death appeared quite low, which may be the result of two hypotheses: cases with a high level of severity or treatment provided was not properly applied. Given the above, it is recommended that the MS of Brazil establishes measures to enhance the quality of SINAN, the investigation of deaths and should improve primary and hospital care.

- EPIDEMIOLOGY [1187] P 889 - VISCERAL LEISHMANIASIS/HIV CO-INFECTION: EPIDEMIOLOGY IN ALAGOAS, BRAZIL, 2001 TO 2012.

CALHEIROS, M.B.; ALVES, W.A.; FERREIRA, I.V.G.; LESSA, M.M.; GOMES, A.R.F.; GOES, J.J.; FREITAS, I.S.; PEIXOTO, V.S.; SOUZA, C.M.; CORTEZ, A.M.C.; SOBRINHO, A.C.R.; MACEDO, D.S.; SILVA, M.E.B.; SÁ, W.T. UNIVERSIDADE FEDERAL DE ALAGOAS, MACEIÓ, AL, BRAZIL.

Keyword:visceral leishmaniasis; hiv; epidemiology

Abstract: Visceral leishmaniasis (VL) still configures as an important neglected disease in Brazil, with significantly magnitude and lethality. In recent decades, co-infection Visceral Leishmaniasis/HIV (VL/HIV) has drawn attention due to the risks already known for complications and deaths. The study aimed to draw a clinical-epidemiologic profile of LV/HIV in Alagoas, northeast region of Brazil, registered in the Information System for Notifiable Diseases (SINAN) and in the Mortality Information System (SIM) of the Brazilian Ministry of Health (HM). This is a cross-sectional and descriptive study. 1168 confirmed cases of VL were recorded in SINAN, from 2001 to 2012. From these ones, 15 (1.28%) were co-infected with HIV. From 2001 to 2010, 02 deaths were recorded in SIM whose underlying cause (disease that initiated the train of morbid events leading directly to death) received the codification of ICD10 (International Classification of Diseases) for LV (B55.0) and terminal/immediate causes for HIV (B24.X). Profile of Individuals LV / HIV in SINAN: 10 (66.6%) male. Age: mean = 30.5 years; most prevalent age group from 20 to 39 years (73.3%). Race: 100% brown. Education: 2 (13,3%) illiterate, 7 (46.6%) elementary school uncompleted and completed, 1 (6.6%) high school. Education: 33% without data recorded. Area of residence: Urban: 11 (73.3%) cases; periurban and rural area: 4 (26,6%) cases. Clinical manifestation: fever (93.3%), weakness and weight loss (86.6%), cough (73.3%), splenomegaly (80%), hepatomegaly (60%), association to infectious conditions, such as meningitis, tuberculosis and tuberculous meningitis (46.6%).

Criterion for LV´s confirmation: Parasitological diagnosis: 9 (60%); Serological diagnosis: 2 (13,3%). Initial treatment: 7 (46.6%) used the pentavalent antimony, 2 (13.3%) amphotericin B. It is noteworthy that only 1 (6.6%) used liposomal amphotericin B and in 4 (26.6%) cases there was no drug administration. Clinical evolution: Healing: 9 (60.0%); Death: 6 (40.0%). The data from the study are consistent with data from Brazilian Ministry of Health. The cases of VL/HIV are mostly young adults, male, brown race, low education and from urban residence area.

Clinical manifestations are compatible with LV. The high mortality rate in these patients is worrisome and the information from this study should support directly actions to primary care and hospital for this type of pacient in the state of Alagoas.

- EPIDEMIOLOGY [1188] P 890 - RISK FACTORS LINKED TO MUCOSAL FORM ON LEISHMANIASIS IN ALAGOAS, NORTHEAST, BRASIL, FROM 2001 TO 2012 PEIXOTO, V.S.; SOBRINO, A.C.R.; GÓES, J.J.; CALHEIROS, M.B.; FREITAS, I.S.; LESSA, M.M.; MACEDO, D.S.; SOUZA, C.M.; CORTEZ, A.M.C.; GOMES, A.R.F.; FERREIRA, I.V.G.; DE SÁ, W.T.; ALVES, W.A. FACULDADE DE MEDICINA - UFAL, MACEIO, AL, BRAZIL.

Keyword:risk; mucosal; leishmaniasis

Abstract: Cutaneous leishmaniasis (CL) is widely spread and presents important morbidity in Brazil and worldwide. The disease has a mean clinical course of 2 months, but it may be shorter or longer (two years) after the bite of an infected sand fly. In Brazil, registration occurs throughout the country and, according to data released by the Ministry of Health (MoH), the mean of the last six years was 20,404 cases / year and 93 deaths happened (mortality rate = 0.08%).

Approximately 93% of the cases had clinical cutaneous form (LTC) and 7% mucosal (LTM).

Brazil’s northeast recorded, in the same period, an average of 4587 cases / year, 22 deaths (mortality: 0.5%), 97% of cutaneous and 3% mucosal forms. Lesions on mucosal form of the disease are less frequent and usually appear after years of skin disease. There are many factors that determine the transmission cycle of LT. However, there are few studies on risk factors for shaping the disease clinical form. Factors that determine the involvement of LTM are not entirely known. Researchers suggest host and parasite’s intrinsic factors are important in LTM.

The objective is to analyze risk factors for LTM through the records in the Information System for Notifiable Diseases MS (SINAN) from 2001 to 2012, of Alagoas, Brazil. Cross-sectional analytical study was conducted. 928 cases of LT were found, 58 cases of LTM(6.3%) and 870 of LTC (93.7%). Gender: 41% male, 59% female (prevalence ratio [PR] = 0.58, 95% confidence interval [95] = 0.35 to 0.95, P = 0.03). Age (years): Average LTM = 44 (Median = 49); LTC = 30 (Median = 25) (p <0.0001). Age (years): <20: 19%, 20: 81% (PR = 3.1, 95 = 1.6 to 5.8, p <0.001). Area of residence: Rural: 55%; Urban: 45% (PR = 0.53, 95 CI = 0.3 to 0.9, p <0.01).

Confirmation criterion: 86% laboratory confirmation, 14% clinical and epidemiological. Clinical outcome: 60% cure, 40% no information. Susceptibility to LT is universal. Disease surveillance aims to reduce mucosal form (severe manifestation) and prevent deformities. Our study highlights the necessity of studies about risk factors for LTM to better understand the disease’s epidemiology in northeastern Brazil.

- EPIDEMIOLOGY [1189] P 891 - STUDY OF LETHALITY BY CUTANEOUS LEISHMANIASIS IN ALAGOAS, 20012012, NORTHEAST, BRAZIL. MACEDO, D.S.; SOBRINHO, A.C.R.; ALVES, W.A.; CALHEIROS, M.B.; FREITAS, I.S.; GOES, J.J.; PEIXOTO, V.S.; LESSA, M.M.; FERREIRA, I.V.G.; SOUZA, C.M.; SÁ, W.T.; GOMES, A.R.F.; SILVA, M.E.B.; PEREIRA, C.W. UNIVERSIDADE FEDERAL DE ALAGOAS, MACEIÓ, AL, BRAZIL.

Keyword:cutaneous leishmaniasis; deaths; epidemiology

Abstract: The Cutaneous Leishmaniasis (CL) should not result in deaths, because when diagnosed and treated early, the disease progresses to clinical cure. However, the data point to an "apparent low lethality", an important reason for being alert. The study aims to describe deaths by CL in Alagoas, with data registered from the Information System for Notifiable Diseases (SINAN) of Ministry of Health, characterizing them by demographic, epidemiological and clinical variables.

This is a cross-sectional and descriptive study from 2001 to 2012. Variables: age, sex, race, education, residence area, type of entry, clinical form, administered drug, prescribed dose, number of ampoules, drug used in case of treatment failure and time between the start of treatment and death. Among the 1003 CL cases reported by SINAN, 10 died. Profile of deaths: male 60%. Age: 30% were aged over 70 years old; 40%: 50-69 and 30%: 30-49. Race: 10% black and 90% brown. Education: 20% illiterate, 10% high school, 20% uncompleted elementary school, 30% ignored, 20% blanked. The residence area was: 50% rural; 50% urban.

All patients were new cases, with 70% of cutaneous form and 30% of mucosa form. In the treatment, 90% had had pentavalent antimony as initially administered drug and 10% had had amphotericin as initially drug administered. The residence area was: 50% rural and 50% urban.

All patients were new cases, with 70% of cutaneous form and 30% of mucosa form. In the treatment, 90% had pentavalent antimony as initially administered drug and 10% had amphotericin as initially drug administered. Prescribed dose: 10% <10 mg/Kg/day/sb5; 30% between 10 and 15; 30% = 15; 10% = 20 and 20% wasn´t informed, including the patient that used amphotericin B, which also has not information on the number of prescribed ampoules.

Number of ampoules: 40% had used 50-60; 20%:30-49; 20%: 2-9. Only three patients had sufficient information to calculate the daily dose; overdosing was not possible to evaluate. Faced with failure of initial treatment, 30% had used other, 30% does not apply, and 40% were not informed. The time elapsed between the start of treatment and death was greater than 25 days in 20% of cases,15-25 in 40%, 7-13 in 30% and below 6 days in 10% of cases.From these data,there is a possibility that the use of antimony had produced toxicity in the majority of deaths or that may had occurred failures in medical monitoring for patients aged over 50 years old and patients with co-morbidities.

- CONTROL PROGRAMS [1211] P 892 - CANINE CALAZAR PREVALENCE IN CEARÁ STATE IN 2012 SOUSA, A.Q.1; CUNHA, J.C.L.1; LIMA, J.W.O.2; MOURA, F.B.P.1; BARROS, A.S.1; CARIOCA, S.M.1; DE SOUSA, L.L.F.1; COSTA, J.R.A.1; PEDROSA, R.B.1; SANTOS, J.L.B.1 1.SECRETARIA DE SAÚDE DO ESTADO DO CEARÁ, FORTALEZA, CE, BRAZIL; 2.UNIVERSIDADE ESTADUAL DO CEARÁ, FORTALEZA, CE, BRAZIL.

Keyword:prevalence; leishmaniasis ; canine

Abstract: Visceral Leishmaniasis is one of seven world endemic whose absolute priority belongs to World Health Organization. Its main reservoir is the dog, being the responsible by maintenance of this parasite in environment. The effective control of this disease requires the diminishing of its prevalence in dog population, which serves as reservoirs to human disease. The objective of this present study was to identify canine visceral leishmaniasis prevalence in Ceará State in 2012. It is about a transversal descriptive study where datum were obtained from monthly reports of reservoirs control sent from municipalities to state. These datum were from the routine control actions and spontaneous demand of population belonged to Leishmaniasis State Control Program of Ceará, being gathered and presented in relative prevalences way. In 2012 Ceará showed canine Calazar in a general prevalence rate of 0,64%. These rates varied from 4,58 to 0,00 I municipalities. Municipalities with ten highest prevalence rates were in order: Maracanaú (4,58%), Tejussuoca (2,56%), Lavras da Mangabeira (2,44%), Nova Olinda (2,18%), Morrinhos (2,14%), São Gonçalo do Amarante (1,53%), Caucaia (1,47%), Fortaleza (1,37%), Tauá ( 1,17), Miraíma (1,10%). The majority of these municipalities are considered as transmission áreas in an intensive and moderate ways, exception only Miraíma which is considered as sporadic.

However, control actions identified a great concentration of canine positivity in municipalities, which showed the greatest rates of incidence of this zoonosis in human, enphasising the necessity of implementing control actions of this endemic in these municipalities.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1218] P 893 - SANDFLIES (DIPTERA: PSYCHODIDAE: PHLEBOTOMINAE) IN FRAGMENTS OF URBAN FOREST AND NATURAL INFECTION OF NYSSOMYIA WHITMANI BY LEISHMANIA INFANTUM CHAGASI IN MATO GROSSO DO SUL STATE, BRAZIL FERNANDES, M.F.1; ISHIMI, C.M.1; OLIVEIRA, A.G.2; DORVAL, M.E.M.C.2; GALATI, E.A.B.3; DOS SANTOS, K.M.1; DA SILVEIRA PERES, L.L.1; OSHIRO, E.T.2; RAIZER, J.1; FERNANDES, W.D.1; NEGRÃO, F.J.1; SANTOS, M.F.C.2; CASARIL, A.E.2; ANDRADE FILHO, J.D.4 1.UNIVERSIDADE FEDERAL DA GRANDE DOURADOS / UFGD, DOURADOS, MS, BRAZIL; 2.UNIVERSIDADE FEDERAL DE MATO GROSSO DO SUL / UFMS, CAMPO GRANDE, MS, BRAZIL; 3.UNIVERSIDADE DE SÃO PAULO / USP, SÃO PAULO, SP, BRAZIL; 4.CENTRO DE PESQUISAS RENÉ RACHOU, FUNDAÇÃO OSWALDO CRUZ, BELO HORIZONTE, MG, BRAZIL.

Keyword:leishmaniasis; vectors; leishmania

Abstract: In Dourados municipality, situated in a transitional region between Atlantic Forest and Brazilian Savanah (Cerrado) - two significant biomes for the epidemiology of leishmaniasis - sandfly vectors of visceral and cutaneous leishmaniasis agents have been found. The preservation of natural environments favors the survival of these insects and reservoirs of Leishmania sp., and human in contact with these ecosystems may become part of the chain of parasite transmission.

This study sought to investigate the sandfly fauna in urban forest ecosystems of Dourados to identify its diversity, abundance and richness species, the monthly distribution of the species, as well as to investigate natural infection by flagellates in females. A total of 10 urban forest areas were sampled with automatic light traps installed once monthly from November 2010 to

October 2011. The females captured were dissected for the investigation of flagellates in their digestive tracts and were then submitted to polymerase chain reaction to identify the Leishmania species. Species abundance was calculated by the Standardized Index of Species Abundance (SISA). The sandfly fauna consisted of 18 species: Brumptomyia brumpti, Brumptomyia cunhai, Brumptomyia galindoi, Brumptomyia pintoi, Evandromyia cortelezzii, Evandromyia lenti, Evandromyia termitophila, Lutzomyia longipalpis, Migonemyia migonei, Micropygomyia acanthopharynx, Nyssomyia whitmani, Psathyromyia aragaoi, Psathyromyia campograndensis, Psathyromyia shannoni, Pintomyia christenseni, Pintomyia misionensis, Pintomyia pessoai and Sciopemyia sordellii. The most abundance species were Ny. whitmani, Br. brumpti, Pa. aragaoi and Pi. pessoai. The greatest frequency (17.8%) of specimens was captured in September. One female of Ny. whitmani was found to be infected with Le. (Le.) infantum chagasi. In conclusion, the study confirmed the presence in fragmentary urban forests of vectors of agents of visceral and cutaneous leishmaniasis. The participation of the Ny. whitmani in the transmission of cutaneous leishmaniasis is known in Brazil and some other South American countries, however, in relation to visceral leishmaniasis, although it has been found naturally infected by the parasite, further investigations are needed to demonstrate its vectorial capacity.

- EPIDEMIOLOGY [1219] P 894 - INSTITUTION OF LIPOSOMAL AMPHOTERICIN B FOR SEVERE CASES OF VISCERAL LEISHMANIASIS: ANALYSIS OF CLINICAL-EPIDEMIOLOGICAL PROFILE OF TREATED PATIENTS EPIDEMIOLOGICAL, BRAZIL, FROM 2005 TO 2006 GOMES, A.R.F.; LESSA, M.M.; CALHEIROS, M.B.; SOUZA, C.M.; FERREIRA, I.V.G.; PEIXOTO, V.S.; ALVES, W.A.; GOES, J.J.; FREITAS, I.S.; SOBRINHO, A.C.R.; SÁ, W.T.; MACEDO, D.S.; CORTEZ, A.M.C. NÚCLEO DE SAÚDE PÚBLICA, FACULDADE DE MEDICINA, UNIVERSIDADE FEDERAL DE ALAGOAS, MACEIÓ, AL, BRAZIL.

Keyword:visceral leishmaniasis; liposomal amphotericin b; death

Abstract: 90% of reported cases of visceral leishmaniasis (VL) in the Americas are located in Brazil.

Currently there is no evidence on the measures of effectiveness in prevention methods of zoonosis. The Ministry of Health has proposed, in 2004, a new treatment program for severe forms of the disease, in which amphotericin B is indicated as first choice in patients with signs of severity. The main objective of the study was to compare the lethality among patients with VL treated with liposomal amphotericin B and treated only with antimonates, provided by the Ministry of Health in 2005 and 2006. This prevalence study was designed using data contained in the instruments of request and the clinical outcomes of these patients. The analysis were done in the software Epiinfo 3.5.4. Of 320 records analyzed, 33 (10.31%) patients evolved to death in 2005 and 2006, respectively 11 (33.33%) and 22 (66.67%). Ages ranged from 8 months to 78 years incomplete. The males (20 cases - 60.6%) were the mean group. The main signs and symptoms were fever, 32 (96.96%), splenomegaly, 30 (90.90%), paleness, 29 (87.87%), hepatomegaly, 26 (78.78%) 16 (48, 48%) and edema, jaundice, 13 (39.39%), malnutrition, 12 (36.36%), vomiting, 9 (27.27%), diarrhea, 5 (15.15%). Of the total, 12 (36.36%) had other symptoms. Of the total, 16 patients (48.48%) occurred in the Northeast region (BA, EC, PB, PI, RN), 15 (45.45%) in the Midwest region (MG, MS). Most cases occurred in children, 10 (30.30%) and farmers, 8 (24.24%). The association between delay in detection of the disease and patients' clinical conditions corroborates to deaths from visceral leishmaniasis. It would be important that health professionals could be trained to diagnose and treat early, choosing which drug best fits the patient's condition. It is relevant that the disease is the same, but the individuals are not. Therefore, the treatment must meet individual needs with regard to drug efficacy and cost.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1225] P 895 - SAND FLY DENSITY REDUCTION IS LESS MARKED IN PRECARIOUS HOUSING AFTER INSECTICIDE THERMAL FOGGING CHAVES, L.F.1; SALDAÑA, A.2; RIGG, C.2; VALDERRAMA, A.2; CALZADA, J.2 1.NAGASAKI UNIVERSITY, NAGASAKI, JAPAN; 2.INSTITUTO CONMEMORATIVO GORGAS, PANAMA, PANAMA.

Keyword:deltametrin; panama; lutzomyia

Abstract: Insecticide thermal fogging (ITF) is a tool to control vector borne diseases. Insecticide application success for vector control has been associated with housing materials and architecture. We conducted a 15 month longitudinal study that included two deltamethrin (6 mg ai/m2) based ITF interventions in 12 of 24 houses at Trinidad de las Minas, a hyperendemic cutaneous leishmaniasis transmission village in western Panama. During the study we followed sand fly (SF) abundance and found a 50 to 80% reduction in SF density in the fogged houses when compared with control houses, while controlling for seasonal changes in SF abundance associated with rainfall. We found some heterogeneities in the reductions, as abundance changed according to SF species, with Lutzomyia gomezi, Lu. panamensis, Lu. dysponeta and Lu. triramula reducing their density between 40% and 90% after ITF, in contrast to Lu. trapidoi whose density increased 5% after the ITF. Differences in the impact of ITF were associated with housing quality, the most precarious houses, i.e., those with features that ease insect entrance, had a disproportionally larger SF abundance, in some cases with an increased domiciliary SF density following the ITF. Our results suggest that Insecticide application success potential to control SF density and Leishmaniasis transmission could depend on housing quality.

- EPIDEMIOLOGY [1229] P 896 - VISCERAL LEISHMANIASIS-HIV CO-INFECTION IN RIO GRANDE DO NORTE, BRAZIL DE LIMA, I.D.1; LOPES, I.D.1; LOPES, X.G.C.2; NASCIMENTO, E.T.1; PEARSON, R.D.3; JERÔNIMO., S.M.B.1 1.UNIVERSIDADE FEDERAL DO RIO GRANDE DO NORTE, NATAL, RN, BRAZIL; 2.SECRETARIA DE ESTADO DA SAUDE PUBLICA DO RIO GRANDE DO NORTE, NATAL, RN, BRAZIL; 3.UNIVERSITY OF VIRGINIA, CHARLOTTSVILLE, UNITED STATES.

Keyword:co-infection; visceral leishmaniasis; hiv

Abstract: Leishmania-HIV co-infection is an emerging health problem worldwide. Infections by Leishmania or HIV can alter host immune responses. Concomitant infection can result in decreased responses to therapy for both pathogens, disease progression and greater severity. The state of Rio Grande do Norte (RN), Northeast region of Brazil, is endemic for Leishmania infantum. Visceral leishmaniasis (VL) has become a disease of periurban areas in Natal and other larger cities during the past 30 years. Moreover, an increase of concomitant VL and AIDS has been observed as previously reported. The Brazilian Minister of Health recommends the performance of HIV testing for all cases of VL, Amphotericin B is the treatment of choice. The objective of this study was to analyze all cases of concurrent VL-AIDS reported in RN between 2007-2012. A descriptive cross sectional study was conducted. A total of 600 cases of VL were reported in Rio Grande do Norte between 2007 and 2012. Of those, 63 (10.5%) were co-infected with HIV (VL-AIDS). Of the 63 VL-AIDS cases, 89% resided in urban areas,and 89% were males, whereas in VL cases without co-infection, 66.8% (n=401) were males. The mean age of VL-AIDS cases was 35 ±16.47 years and for VL cases without HIV 24.0±18.5 years. The clinical presentations of VL-AIDS and VL without co-infection were similar. Of the 63 VL-

AIDS cases, 36.5% were treated with pentavalent antimonial. However, 14% of the VL-HIV relapsed, 9.5% (n=6) died with progressive VL and 12.7% (n=8) because of other causes. This report confirms that co-infection with HIV in areas endemic for Leishmania infantum has become an important risk factor for VL. Screening for HIV should be performed in all VL cases, principally considering that malnutrition and other risk factors for progressive VL have decreased due to improvement in social-economic conditions and general health factors including routine childhood vaccination - EPIDEMIOLOGY [1230] P 897 - CLINICAL AND EPIDEMIOLOGICAL PROFILE OF AMERICAN CUTANEOUS LEISHMANIASIS (ACL) IN ALAGOAS, NORTHEASTERN BRAZIL. SOUZA, C.M.; DE SÁ, W.T.; FERREIRA, I.V.G.; PEIXOTO, V.S.; GOES, J.J.; ALVES, W.A.; LESSA, M.M.; CALHEIROS, M.B.; FREITAS, I.S.; CORTEZ, A.M.C.; R SOBRINHO, A.C.; MACEDO, D.S.; GOMES, A.R.F.; BEZERRA, M.E. UFAL, MACEIÓ, AL, BRAZIL.

Keyword:cutaneous leishmaniasis; clinical; epidemiological

Abstract: Initially considered a wild enviroment zoonosis, the ACL is becoming a public health problem that affects the countryside and periurban zone. According to DATASUS’s data, in the period from 1990 to 2010, the ACL had an incidence of 20.4 in northeastern Brazil and Alagoas represented 3.34 of the reported cases. The objective of this study is to collect data from clinical and epidemiological aspects of ACL infections in the state of Alagoas, northeastern Brazil, in the period from 2007 to 2012. This study has a descriptive and quantitative approach, based on secondary data obtained from confirmed and registered ACL cases and recorded in the de Secretary of State for Health of Alagoas (SESAU). The study period was from 2007 to 2012.

Besides annual frequency, we collected variables by areas (urban and rural), gender (male and female), race (white, black, yellow, brown and indigenous), clinical presentation (cutaneous or mucosa) and evolution of cases (if there was no cure, abandon, death by ACL, death from other causes or diagnostic changes).Over this period, 476 cases of ACL were registered in Alagoas, with the largest number of cases registered in 2007 ( 121 cases) and 2008 (109 cases). The countryside prevailed, with 80% of registered cases (369) versus 19.1% (87) in the urban area.

Gender prevalence evaluation shows a higher frequency in males with 321 cases, against 155 in females. In respect to race, brown corresponded to 79.5% of the total, with 368 cases against 59 White, 13 black, 13 indigenous and 21 ignored cases. New cases accounted for almost all the cases of input type, with 426 (97.1%) cases and 12 (2.5%) cases of recurrence only, and 2 (0.4%) unrecognized. The predominant clinical presentation was cutaneous, with 460 (96.6%) cases versus 16 (3.4%) mucosa. Cure evolved into a total of 389 (96.6%) cases. In the remaining 3 cases (0.7%) progressed to abandonment, 3 (0.7%) who died of LTA, 1 (0.2%) who died of other causes, and 6 (1.5%) developed change diagnostics. It is clear that male, brown, rural residents correspond to the major risk factors associated with LTA. The cutaneous form maintains the prevalence over the mucosa. Despite still being recorded a significant number of cases within 4 years, fortunately, the vast majority resulted in cure, which suggests the presence of effective medical care.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1240] P 898 - SPECIES RICHNESS AND DIVERSITY OF SAND-FLIES (DIPTERA: PSYCODIDAE: PHLEBOTOMINAE) IN THE INFLUENCE AREA OF THE HYDROELECTRIC POWER PLANT OF JIRAU IN PORTO VELHO, RONDONIA, BRAZIL SILVA, C.L.1; KOBS, T.L.1; GOMES, B.1; RODRIGUES, M.S.2; SILVA, A.A.3; GIL SOARES, L.H.1

1.IPEPATRO, PORTO VELHO, RO, BRAZIL; 2.UNIVERSIDADE FEDERAL DE MINAS GERAIS, PORTO VELHO, RO, BRAZIL; 3.UNIVERSIDADE FEDERAL DE RÔNDONIA, PORTO VELHO, RO, BRAZIL.

Keyword:amazon; ecology; insect monitoring

Abstract: The neotropical fauna of Phlebotominae includes over 480 species and 200 are found in Brazil.

The Brazilian government program for the economy acceleration named PAC includes the construction of hydroelectric power plants (HPP) that might be related to ecological and behavioral changes in the sand fly fauna. Therefore, the present work includes a species inventory and a longitudinal study within the influence area of the construction of the HPPJirau. Sand flies were collected using Shannon and CDC traps during 12 hours (06:00 am – 06:00 pm) in 14 sites during 2011 and 2012. Insects were screened and identified according to Yung & Duncan (1994) and diversity was calculated using Shannon diversity index and compared using ANOVA. 5948 sand flies were collected comprising 58 species and three genera. Lutzomyia amazonensis, Lutzomyia antunesi, Lutzomyia chagasi and Lutzomyia umbratilis comprised 40% of the species collected. These species were previously described as potential vectors of Leishmania spp and represent a potential risk to workers living in the forest areas close to natural reservoirs of this parasite. Despite of that, there were no differences in the diversity index calculated using data from several insect collections during 2011 (1.40 ± 0.20) and 2012 (1.74 ± 0.08) (ANOVA F1,23= 2.30, p=0.14). The HPP-Jirau electrical power generation has already started but the flooding of areas that might affect potential breeding sites of phlebotomine is not complete yet. Thus, new data including natural infection by Leishmania and fauna structure after HPP-Jirau is fully operational will provide further information on the impact of this HPP in the vector-parasite dynamics.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1243] P 899 - THE SANDFLY FAUNA OF THE COFFEE-PRODUCING REGION OF COLOMBIA CONTRERAS, M.A.1; PORTER, C.H.2; VELEZ, I.D.1; URIBE, S.I.3 1.PECET, UNIVERSITY OF ANTIOQUIA, MEDELLIN, COLOMBIA; 2.BCENTERS FOR DISEASE CONTROL AND PREVENTION (CDC), ATLANTA, UNITED STATES; 3.PECET, UNIVERSITY OF ANTIOQUIA AND NATIONAL UNIVERSITY OF COLOMBIA, MEDELLIN, COLOMBIA.

Keyword:leishmania vectors; colombian coffee areas; phlebotominae, taxonomy

Abstract: Coffee Axis region is considered a biological corridor and support a rich diversity of plant and animal species, including phlebotomine sandflies (Diptera: Psychodidae: Phlebotominae), some of which transmit Leishmania to humans. The study of the insect vectors in coffee areas is critical for monitoring and prediction of leishmaniasis; a vector-borne diseases of public health importance. The purpose of this study was to obtain information of phlebotomine sand flies in coffee-producing region of Colombia. Sand flies were captured and identified to species.

Sandflies were sampled between the 2009 and 2012 at night using Shannon trap, CDC light trap and mouth aspirator on human and animal bait. Day-time resting sites were sampled by mouth aspirator. Taxonomic identification was performed according to Young and Duncan (1994) and Galati (2010) keys. A total of 1630 sand flies (1.013 females and 617 males) were collected from nine localities investigated. All collected sandflies were identified as belonging to one of 29 species and were collected from nine localities investigated: Alcalá (25 specimens), Jericó (9 specimens), Jardín (87 specimens), Pueblo Rico (250 specimens), Quimbaya (3 specimens), Chinchiná (236 specimens), La Victoria (141 specimens), Samaná (195 specimens) and Salamina (685 specimens). With 526 (35.21% %) specimens, Lutzomyia pia was the specie most frequently collected, followed by Lutzomyia carrerai thula (12.94%), Lu. yuilli (6.69%), Lutzomyia youngi (6.26%), Lutzomyia columbiana (6.07%) y Lutzomyia panamensis (4.72%).

Seven highly anthropophilic sand fly species that are suspected vectors of the parasites were identificated. Thus, these species may be implicated in disease transmission in the studied area.

In our study, we report new departmental records of phlebotomine sandflies in the department of Caldas: Lutzomyia carpenteri, Lutzomyia dasymera, Lutzomyia walkeri, Lutzomyia carrerai thula and genus Brumptomyia. In addition, Brumptomyia beaupertuyi y Lutzomyia sp. de Baduel are new records in the department of Antioquia. The geographic distribution of the genus Lutzomyia is being greatly extended to new Colombian localities in coffee areas in the Andean region. The necessity of systematic and taxonomic studies to know the role of these insects in the transmission of pathologies affecting humans is emphasized.

- EPIDEMIOLOGY [1247] P 900 - RELATIVE RISK OF VISCERAL LEISHMANIASIS IN AN ENDEMIC AREA IN BRAZIL MARQUES, L.H.S.1; ARAĂ&#x161;JO, V.E.M.2; REIS, I.A.3; ROCHA, I.C.M.2; VITAL, W.C.4; PFEILSTICKER, T.R.1; MORAIS, M.H.F.2; CARNEIRO, M.5 1.UFMG- ICB, BELO HORIZONTE, MG, BRAZIL; 2.PREFEITURA DE BELO HORIZONTE, BELO HORIZONTE, MG, BRAZIL; 3.UFMG- ICEX, BELO HORIZONTE, MG, BRAZIL; 4.UFMG, BELO HORIZONTE, MG, BRAZIL; 5.UFMG-ICB, BELO HORIZONTE, MG, BRAZIL.

Keyword:relative risk ; visceral leishmaniasis ; bayesian approach

Abstract: Understanding of relative risk (RR) of visceral leishmaniasis (VL) according to small-areas may assist in the planning of the VL Control and Surveillance Program (VL-CSP). Therefore, it is important to focus on areas of greatest risk disease in urban environment. The aim of this study was to estimate the relative risk for LV in 147 coverage areas (CA) of the basic health units in the city of Belo Horizonte, in the period of 2007-2011. The Bayesian approach was used to implement models to estimate the relative risk for LV, using the software WinBUGS 1.4 and considering the spatial dependence between neighboring areas. This spatial statistical modeling minimizes the instability of crude rates resultant from the low frequency of cases in small-areas.

The human VL cases (n = 646) and its distribution among the areas were the parameters used to estimate the RR, as well as the population of these areas. The results showed that there is a transition of the areas in relation to risk with clusters of high risk in certain health districts. In the period of the study, 44% of low-risk areas become medium-risk areas. Amongst the areas initially classified as high-risk (RR 1.53), 57.6% were finally classified as medium risk-areas.

At baseline, half of the areas of high-risk was concentrated in the health districts Northeast (11) and Venda Nova (2). In the course of time, all areas of high-risk remained localized in the same districts. Despite the clusters of high-risk areas, we found great variation between the RR areas located in the same health district. Therefore, we observed a reduction of clusters of high and low incidences, while there was an increase in the number of that medium-risk area, suggesting a dynamic transmission pattern of the disease. Thus, the estimation and monitoring of RR of VL according to small-areas is important to support the planning, evaluation and orienting measures of the VL-CSP in an urban environment, especially in the large cities.

- OTHER SPECIAL TOPICS [1249] P 901 - MODELING NEGLECTED TROPICAL DISEASES IN COLOMBIA AND BOLIVIA NIETO, P.D.1; MALONE, J.B.1; VONATSOU, P.2; MCCAROLL, J.3

1.LOUISIANA STATE UNIVERSITY, BATON ROUGE, UNITED STATES; 2.2SWISS TROPICAL AND PUBLIC HEALTH INSTITUTE, BASEL, SWITZERLAND, SWITZERLAND; 3.LOUISIANA STATE UNVERSITY, BATON ROUGE, UNITED STATES.

Keyword:gis; neglected tropical diseases; niche modelling

Abstract: In Latin America and the Caribbean neglected tropical diseases NTDs are the most common infections of approximately 200 million of the poorest people in the region (Hotez et al, 2008).

NTDs primarily occur in the poorest and the most marginalized communities where unsafe water, poor sanitation and limited access to basic health care are rife. Control efforts require reliable baseline maps of the geographical distribution of the disease burden and population at risk to allow better targeting of planned interventions at the high risk areas. This work integrates compiled disease from ministries of health, research institutions and publications with environmental, climatic and socioeconomical factors to create statistical models to identify the most significant predictors of disease transmission and provide a mathematical description of the disease-predictors that can be used for control purposes. The maximum entropy was used for generating probability surfaces of the geographical distributions of NTD. The final results were models that predict areas where the diseases are most likely to occur based on environmental correlations. Ecological niche modeling using Maxent focused on vectors of Chagas disease and diseases data on Chagas and leishmaniasis in Colombia as prototypes to develop methodologies to be used for model development for other NTDs of interest in Bolivia and Brazil.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1255] P 902 - PHLEBOTOMINE SAND FLY FAUNA (DIPTERA: PSYCHODIDAE) AND NATURAL FLAGELLATE INFECTIONS IN AMERICAN CUTANEOUS LEISHMANIASIS ENDEMIC AREA UNDER THE INFLUENCE OF BELO MONTE HYDROELECTRIC PLANT AT STATE OF PARÁ, AMAZONIAN BRAZIL. DA SILVA, F.M.M.; DOS SANTOS, T.V.; SILVA SANTOS, L.A.; LEÃO, E.F.; BRANDÃO, R.C.F.; SILVEIRA, F.T. INSTITUTO EVANDRO CHAGAS, ANANINDEUA, PA, BRAZIL.

Keyword:phlebotomine; fauna; hydroelectric plant

Abstract: Background: The increase in American Cutaneous Leishmaniasis (ACL) cases has been related to the deforestation of primary forests, leading to environmental changes and imbalances such as construction of dams and hydroelectric plants. The present study aims to collaborate with the eco-epidemiological knowledge about transmission of ACL presenting the sand fly fauna and natural flagellate infections in an endemic area under the influence of Belo Monte hydroelectric plant at state of Pará, Amazonian Brazil.

Methods: During three 15-day entomological excursions in February, July and October 2012, nine selected areas of capture in four municipalities under direct influence of the plant were investigated as follow: Altamira, Vitória do Xingú, Senador José Porfírio and Anapu. Captures were undertaken with 10 CDC light traps (9 at 1,5 meters to ground and 1 at 20 m in tree canopy) disposed from 6:00 p.m. to 6:00 a.m. and Shannon traps from 6:00 to 8:00 p.m. Sand flies were washed in saline solution to dissect the females and search natural flagellate infections. Females infected had their digestive tract inoculated into Difco B45 media. Males were identified and stored in alcohol 70%.

Results: A total of 3705 individuals (1382 males and 2323 females) distributed among to 51 species were captured. The most frequent species were Lutzomyia davisi (23,4%) and Lu. complexa (11,9%). Natural flagellate infections were found in 2 specimens of Lu. davisi, 1 from Lu. evandroi and 1 from Lu. walkeri. Unfortunately, all positive flies did not succeed in cultivating flagellates.

Conclusion: These data reinforce the importance to investigate Lu. davisi as a vector of ACL in Amazon region once its highly anthropophilic behavior, as well as it has previously been found infected with Leishmania (Viannia) spp. in several occasions at Amazonian Brazil.

Financial support: Norte Energia S.A./ Instituto Evandro Chagas - EPIDEMIOLOGY [1260] P 903 - INTRASPECIFIC GENETIC VARIABILITY OF LEISHMANIA (LEISHMANIA) INFANTUM IN SAMPLES FROM DOGS AND HUMANS DA SILVA, T.A.M.1; GOMES, L.I.2; OLIVEIRA, E.J.2; VITAL, W.C.1; KER, H.G.3; REIS, A.B.3; RABELLO, A.2; CARNEIRO, M.1

1.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 2.CENTRO DE PESQUISAS RENĂ&#x2030; RACHOU - FIOCRUZ, BELO HORIZONTE, MG, BRAZIL; 3.UNIVERSIDADE FEDERAL DE OURO PRETO, OURO PRETO, MG, BRAZIL.

Keyword:leishmania (leishmania) infantum; intraspecific variability; pcr-rflp

Abstract: Visceral leishmaniasis (VL) is a disease distributed worldwide and potentially fatal to humans if not diagnosed and early treated. In addition to human, other mammals are also affected, especially dogs in the urban context, which are the major reservoir of the disease. Individuals showing human immunodeficiency virus (HIV) infection are also susceptible to VL and deserve more attention. The city of Belo Horizonte, Minas Gerais state, has one of the highest lethality rates for VL in humans in Brazil and a high prevalence for the disease in dogs. Leishmania (Leishmania) infantum is the etiological agent for VL in Brazil and the aim of this work was to study the intraspecific genetic variability of Leishmania from human and dogs cases in Belo Horizonte, and some other municipalities in Minas Gerais state, applying PCR-RFLP with different DNA targets: kDNA and gp63. In this work were evaluated 47 DNA samples extracted from clinical specimens and L. infantum culture. These DNA samples were amplified for kDNA using the primers MC1 and MC2, which were specific to the Leishmania donovani complex and generate a fragment of 447bp (base pairs). For gp63 glycoprotein the primers used were SG1 and SG2, which generate a fragment of 1330bp. To perform the kDNA RFLP five restriction enzymes were used: VspI, RsaI, BglII, DdeI and Hpa II. The gp63 glycoprotein RFLP was performed with two enzymes: Hinc II and TaqI. The results obtained with RsaI, BglII, DdeI, Hpa II, HincII and TaqI restriction enzymes proved to be monomorphic. Two distinct profiles were identified with the VspI enzyme: Type I (revealed by 46 DNA samples) and Type II (shown for only one DNA sample evaluated in the study). This same result was also seen by phenogram analysis, constructed with binary matrix and neighbor-joining clustering. The variable sample was from an adult who presented classic VL and is resident of Minas Novas city, Minas Gerais state, municipality not belonging to the Metropolitan Region of Belo Horizonte. These findings reveal there is a high degree of homogeneity among Leishmania from human and dogs in Belo Horizonte, and some other municipalities in Minas Gerais state, Brazil.

- CONTROL PROGRAMS [1261] P 904 - ECO-EPIDEMIOLOGICAL ANALYSIS OF CUTANEOUS LEISHMANIASIS TRANSMISSION IN THE NORTHWESTERN CARIBBEAN COAST REGION OF COLOMBIA CARRILLO, L.M.; CADENA, H.; ACOSTA, L.A.; MONDRAGON, K.; VELEZ, A.; AGUDELO, D.; PEREZ, J.E.; TRUJILLO, J.J.; VELEZ, I.D. PECET, UNIVERSITY OF ANTIOQUIA, MEDELLIN, COLOMBIA.

Keyword:ecoepidemiology; sand fly vectors; cutaneous leishmaniasis

Abstract: The eco-epidemiological method applied to a Cutaneous Leishmaniasis (CL) focus implies a holistic approach where the biological, exact, social and health sciences are joined in order to have a precise comprehension of the natural history of transmission and of the epidemiological infection risk. The main variables in the study are summarized as follows: transmission area, season of greatest risk, hours of higher vector bite activity, characteristics of population in higher risk of infection and place, regarding to domicile, where most people is infected. This information is needed for the design and implementation of affordable and effective control and prevention programs.

This study was carried out during one year, in the Northwestern Caribbean Coast region of Colombia, borders with Panamá in an area of approx. 500 Km2 where 3 natural regions are clearly differentiated: Sylvatic, rural and urban. The performed activities were: 1) Entomological cross sectional study, 2) Active search of cases 3) epidemiological survey using Montenegro skin test, 4) search of domestic and wild animals as possible reservoir hosts, 5) Study of knowledge, attitudes, and practices (KAP) and 6) primary health education for the community.

A total of 4763 phlebotomine sandflies belonging to twenty-five species were collected. Lu. panamensis was the most abundant species, it is present in the 3 natural regions and it was captured in intra, peri and extra domicile, its bite activity starts at 6 p.m. Its high density and the background of being vector in other regions of Colombian allow us to incriminate it as the main vector of Leishmania spp in this region. It is more abundant during the raining season which represents the highest transmission risk.

Lu. cayennensis cayennensis, incriminated as a vector in Panama, was also captured and it`s a new record for this region, although its density was low.

Thirty six patients with CL and 1 with MCL were found. Thirteen strains isolated were identified as L. panamensis. Patients were treated successfully with Miltefosine and a complete cure in all of them was recorded.

111 wild animals from nine species were capture. Four of them were found naturally infected with L. panamensis : Didelphis marsupialis (6/67), Proechymis semispinosus (3/18), Metachyrus nudicaudatus (1/7) and Choloepus hoffmani (1/1). One dog (Canis familiaris) with scrotal lesion was also positive for L. panamensis. D. marsupialis is proposed as liaison host from sylvatic to urban areas and C. familiaris as an accidental host.

In general, the population does not recognize the sandflies as vectors and don’t use antivectorial control measures. Many inhabitants belong to the Evangelic church and they don´t request the health services when they acquire the disease due to their beliefs. There is a high transmission rate in the 3 regions, even though it is higher in the rural area. In the urban area children are the most infected population.

- EPIDEMIOLOGY [1263] P 905 - IS LEISHMANIASIS WIDESPREAD IN TUNISIA ? : FIRST DATA ON LEISHMANIA KILLICKI IN THE PROVINCE OF GAFSA, SOUTHWEST TUNISIA. JAOUADI, K.1; HAOUAS, N.1; GORCII, M.1; KIDAR, A.2; PRATLONG, F.3; DEDET, J.3; BABBA, H.1 1.UNIVERSITÉ DE MONASTIR, MONASTIR, TUNISIA; 2.HOPITAL RÉGIONAL DE GAFSA, GAFSA, TUNISIA; 3.UNIVERSITE DE MONTPELLIER 1, MONTPELLIER, FRANCE.

Keyword:leishmania killicki; cutaneous leishmaniasis; gafsa, tunisie

Abstract: In Tunisia, four clinico-epidemiological forms of leishmaniasis have been recognized: Chronic anthroponotic cutaneous leishmaniasis (CL) encountered in a microfocus in the southeast of the country and caused by Leishmania killicki; Zoonotic CL, epidemic in the centre and south of the country, caused by Leishmania major; Sporadic CL in the north caused by Leishmania infantum; and visceral leishmaniasis (VL) in the north and the center caused by Leishmania infantum. Gafsa area in the South-west of Tunisia is a classical focus of CL due to Leishmania major. We report thirty human cases due to Leishmania killicki, affecting men and women from 3 to 77 years old in area where the disease was previously unknown. Leishmania killicki have been typed using molecular techniques: polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) and gene sequencing. Ten strains from patients have been successfully cultured on NNN medium and isoenzymatically typed as Leishmania killicki MON-8. Our result strongly suggests that Gafsa is a new Leishmania killicki focus with a stable transmission cycle. Cases are considered as autochthonous and are the first to be published in this region of Tunisia.

- RESERVOIRS [1266] P 906 - DETECTION OF LEISHMANIA (VIANNIA) SPP IN WILD AND SYNANTHROPIC RODENTS CAPTURED IN AN ENDEMIC AREA OF AMERICAN CUTANEOUS LEISHMANIASIS IN THE ZONA DA MATA OF PERNAMBUCO, BRAZIL. MARINHO-JUNIOR, J.F.1; BRITO, L.P.S.1; CARVALHO, A.W.S.1; MORAES, R.C.S.1; CARVALHO, F.G.1; CAVALCANTI, M.P.1; COURTENAY, D.A.O.2; BRANDAO-FILHO, S.P.1 1.CENTRO DE PESQUISAS AGGEU MAGALHﾃウS - CPQAM/FIOCRUZ, RECIFE, PE, BRAZIL; 2.UNIVERSITY OF WARWICK, WARWICK, UNITED KINGDOM.

Keyword:reservoir; american cutaneous leishmaniasis; leishmania (viannia) braziliensis

Abstract: Introduction: Leishmaniasis, caused by different species of protozoa of the genus Leishmania are antropozoonoses, considered as a major public health problem in the world, they represent a complex of diseases with epidemiological diversity, where they participate in transmission cycles, various etiological agents, vectors and hosts. In the Northeast the predominant species involved with the American Cutaneous Leishmaniasis (ACL) is Leishmania (Viannia) braziliensis. One of the main shortcomings related to eco-epidemiology of ACL associated with Leishmania (Viannia) braziliensis is related to the identification of reservoir hosts and sand fly that keeps the transmission cycle of the parasite. The municipality of Amaraji, located in the Zona da Mata of Pernambuco South, has an important incidence of ACL. Objective: Detection of natural infection of Leishmania (Viannia) spp in wild animals and synanthropic, to identify reservoir hosts involved in maintaining zoonotic cycle in this region. Material and Methods: the months of may and december 2012, there were catches of wild and synanthropic in the town of Raiz de Dentro, Amaraji municipality, located in the Zona da Mata of Pernambuco. The captured animals were anesthetized, marked with microchip, collected skin samples (right ear) and blood, and later returned to the field at the capture site. The samples will be subjected to DNA extraction and purification, and analyzed by real-time PCR for detection of natural Leishmania infection. Results: We collected 193 animals wild and synanthropic belonging to 8 different species, of which 107 males and 86 females. The predominant species in the catch were Rattus rattus (59/193) and Nectomys squamipes (53/193), corresponding to 30.6% and 27.5% respectively of the total. We analyzed 100 samples of DNA from blood, where 25 (25%) were positive. Conclusions: The detection of natural Leishmania infection in animals enhances

their participation in the transmission cycle of ACL. The continuity of the approach of this study, together with infectivity assays by xenodiagnosis with sandflies can reinforce the hypothesis of activity wild and synanthropic rodents as reservoir hosts of ACL primary or secondary. Support: CAPES, CNPq and FACEPE. - EPIDEMIOLOGY [1267] P 907 - EPIDEMILOGY OF VISCERAL LEISHMANIASIS IN THE STATE OF MARANHÃO, BRAZIL: REPORT OF ACUTE CASES FROM 2001 TO 2011.

SILVA, L.A.C.1; COSTA, M.A.2; DOS REIS, A.S.1; SILVA, L.C.1; PAREDES, A.O.1; CAMELO, N.L.1; GUERRA, R.N.M.1 1.UFMA, SAO LUIZ, MA, BRAZIL; 2.IFES, SAO LUIZ, MA, BRAZIL.

Keyword:visceral leishmaniasis; prevalence; maranhão

Abstract: Leishmaniasis is a complex of several diseases caused by protozoa of the genus Leishmania, with more than 350 million people are at risk of contracting some form of the disease.

According to the Ministério da Saúde do Brasil, Maranhão, Ceará, Piauí and Bahia are the Brazilian states with the highest numbers of cases of visceral leishmaniasis in recent decades.

Therefore, the objective of this study was to investigate the prevalence of visceral leishmaniasis in the state of Maranhão in the last 11 years (2001 to 2011). Therefore, we evaluated the data obtained in Sistema de Informação de Agravos Notificáveis (SINAN). We evaluated the total number of confirmed cases by region, middle region, gender and ethnicity. Between the years 2001 to 2011, there were 5624 confirmed cases of visceral leishmaniasis throughout Maranhão.

The year 2003 had the highest number of cases, with 14.4% of cases, and the year of 2010 the lowest with 7.2%, a decrease of 50%. The LV predominates in urban Maranhão, totaling 3925 patients confirmed in 11 years, equivalent to 70%, above the half, well above the total cases of all other zones. Surpassing even the rural area (25.4%). The city of San Luis has the largest number of confirmed cases of VL periurban area of the state, with 41.5% (N = 39) of cases. In the period from 2001 to 2011 males were more affected than females accounting for 58.8% of cases, with 960 patients diagnosed men more than women. Independent zone or middle region, Maranhão is made up of people from various ethnic groups, generating a large amount a people of mixed race, reflecting the number of VL cases in relation to ethnicity that affects mainly people of mixed ethnicity (75.2%). With respect to population, there is a worrying fact, the year 2011 has the highest mortality rate of LV in Maranhão the past five years, with an incidence rate of 6.6 / 100,000 inhabitants., A fact that reflects the current situation and demonstrates that the disease is not controlled in the state. These data show the factors and conditions that contribute to the onset of the disease and, consequently, there is the situation and evolution of the disease in the state in recent years, the diagnosis in humans and epidemiological knowledge is necessary to control of this disease.

- RESERVOIRS [1271] P 908 - CUTANEOUS LEISHMANIASIS IN TUNISIA: STUDY OF THE DISEASE RESERVOIR AND FIRST DETECTION OF LEISHMANIA KILLICKI (KINETOPLASTIDA, TRYPANOSOMATIDAE) IN CTENODACTYLUS GUNDI (RODENTIA, CTENODACTYLIDAE) JAOUADI, K.1; AMOR HAOUAS, N.1; GORCII, M.1; BOUDABOUS, R.1; DHAHRI, B.2; PRATLONG, F.3; DEDET, J.P.3; SLIM, E.2; BABBA, H.1 1.UNIVERSITÉ DE MONASTIR, MONASTIR, TUNISIA; 2.DIRECTION DES SOINS DE SANTÉ DE BASE, GAFSA, TUNISIA; 3.UNIVERSITÉ DE MONTPELLIER 1, MONTPELLIER, FRANCE.

Keyword:leishmania killicki; ctenodactylus gundi; tunisia

Abstract: In Tunisia, Leishmania infantum, L. major and L. killicki are responsible for cutaneous leishmaniasis (CL). This last taxon has been described in 1980 in Southeast country; it was also recently reported in Lybia and Algeria. Nevertheless, neither vector nor reservoirs of this parasite are known. The identification of the vector and the animal reservoir host of L. killicki is critical for the establishment of an efficient control strategy. Blood, popliteal lymph node, spleen, bone marrow, liver and skin were collected from 50 rodents in 2009 in south western Tunisia. Samples were smeared onto glass slides, cultured on NNN medium and tested by polymerase chain reaction for Leishmania detection. Parasites were detected by PCR from 10 Psammomys obesus and from two Ctenodactylus (C.) gundi. Parasite identification was performed simultaneously by internal transcribed spacer 1 PCR-RFLP and by PCR sequencing.

Both L. major and L. killicki were identified from infected Psammomys and C. gundi respectively. This is the first report of L. killicki identified from C.gundi in Tunisia. This result supports the assumption that C. gundi is a potential reservoir for L. killicki. However, the isolation of the parasite from this rodent is crucial for the confirmation of this first result.

Further investigation, such collecting wild rodents in other L. killicki foci, is required to discern the potential epidemiologic role of C. gundi in spreading infection.

- EPIDEMIOLOGY [1285] P 909 - EPIDEMIOLOGY OF MUCOCUTANEOUS LEISHMANIASIS IN VENEZUELA DE LIMA, H.; BORGES, R.; BELIZARIO, D.; CONVIT, J. INSTITUTO DE BIOMEDICINA, CARACAS, VENEZUELA; .

Keyword:mucocutaneous leishmaniasis; epidemiology; venezuela

Abstract: Mucocutaneous leishmaniasis (MCL) is an aggresive form of the disease, fortunately not very frequent, which prevails in the americas affecting mainly socially excluded populations. The objective of this study was to carry out a retrospective descriptive analysis of MCL in Venezuela for the period 1991-2010. A total of 491 cases were registered. During this period, a decreasing tendency was observed in the number of cases per year, with a mĂ­nimum of 10 in 2007 and a mĂĄximum of 48 in 2000. MCL represents approximately 1% of the total amount of cutaneous leishmaniasis cases reported annually in Venezuela, and reflect rates per 100,000 inhabitants that vary between 0.04 and 0.20. It is predominant in the male genre with a male/female ratio of 2.25 (340/151). In the Bolivar State, a statistically significant increase of this ratio was observed with 5.23 (p < 0.01), while in the Lara State this ratio is equal to 1.18 (p < 0.05). The average age at the time of the diagnosis was 44.67 years and even though cases are observed in all ages, there is an increase with age both in number and rate. Almost one third (31.29%) of the cases are farmers (143/457), household work 20.35% of the cases, and others (including mining) represent 16.85%. Illiteracy is very high 34.73%; difference with the national average is statistically significant. Positive leishmanin skin tests (LST) in 97.59% of the cases, and of these 34.46% are considered hyperreactive. (LST 30 mm). According to the place of infection, 77.80% of the cases (382/491) originating in seven states (Barinas, Bolivar, Lara, Merida, Portuguesa, Tachira and Zulia), are also distributed in 11.82% (132/1,116) of the totality of parishes at national level. These States and their respective parishes are associated almost entirely with rural populations located in mountain ranges such as: the Andes, Serrania de Perija, Andes Foothills or the Guayana Highlands. However, an important group of cases have as probable place of infection urban centers of the Lara State. Conclusions: (i) MCL is not very frequent in Venezuela and appears mainly in certain rural parishes that may be identified well. (ii) There appears to exist different transmission patterns in rural and urban areas. In general in rural areas, it is more common in men probably given to their association with certain activities

(farming and mining), while in urban areas (some parishes of the Lara State) tend to affect men and women equally. (iii) The high percentages of illiteracy found in the affected groups, evidence the existing conditions of social exclusion and inequality. (iv) Although LST is not specific for the diagnosis of MCL it is highly sensitive for their detection, and given to the high proportion of hyperractors could serve as a guide in the diagnosis.

- EPIDEMIOLOGY [1286] P 910 - ECO-EPIDEMIOLOGICAL CHARACTERIZATION OF A CUTANEOUS LEISHMANIASIS FOCUS IN THE AVILA MOUNTAIN, CARACAS, VENEZUELA ZERPA, O.1; CADENA, H.2; VIVEROS, R.J.2; BELIZARIO, D.3; GALINDO, W.3; MORALES, J.3; FELICIANGELI, M.D.4; VELEZ, I.D.1 1.INSTITUTO MEDICO LA FLORESTA, CARACAS, VENEZUELA; 2.PECET, UNIVERSITY OF ANTIOQUIA, MEDELLIN, COLOMBIA; 3.INSTITUTO DE BIOMEDICINA, CARACAS, CARACAS, VENEZUELA; 4.UNIVERSIDAD DE CARABOBO, FACULTAD DE CIENCIAS DE LA SALUD, BIOMED, MARACAY, VENEZUELA.

Keyword:ecoepidemiology; san flies vector; cutaneous leishmaniasis

Abstract: In June 2012 a cross-sectional epidemiological study was performed in the municipality of San JosĂŠ de Galipan, an endemic area for CL not previously studied, located at the Avila mountain, close to Caracas city.

The purpose of the study was to train local health staff on foci studies and determination of risk of infection with Leishmania spp . in support the leishmaniasis control program.

Active search of cases, an eco-epidemiological survey by Montenegro Skin Test (MST), phlebotomine sandfly sampling using CDC traps indoors and in the peri-domestic habitat as well as Shannon traps in the woodland and captures in resting sites, and clinical evaluation of domestic animals looking for possible leishmaniasis lesions, were also implemented.

MST was applied to 106 people. The positivity was 58% without difference by gender. In children under 14 years old, the positivity was 35%. The 40% of positive MST people had scars and clinical history of cutaneous leishmaniasis. No active CL or mucosal leishmaniasis cases were found.

Preliminary results showed the presence of six sandfly species: L. ovallesi , L. lichyi, L. migonei, L. cayennensis, L. atroclavata and L. dubitans. L. ovallesi was the most abundant species. ITS1 nested PCR was used to search for Leishmania spp. infection, which so far was revealed in L. ovallesi and L. lichyi. No domestic animals with CL lesions were found.

In conclusion this is an endemic area for CL, where transmission occurs mainly in the intra and peri-domicile as no difference in positive MST was found by ages group or gender. Lu. ovallesi is incriminated as the main vector as it is the most abundant species and was found naturally infected with Leishmania spp. The absence during the survey of active human or animal cases made difficult the strain isolation for further identification.

Acknowledgments: This project was performed thanks to the financial support of SanofiAventis.

- RESERVOIRS [1291] P 911 - TRIPANOSOMATIDAE INFECTION ON WILD SMALL MAMMALS FROM THE CAPIVARI RIVER BASIN, MIDDLE EAST OF SÃO PAULO STATE, BRAZIL. CUTOLO, A.A.1; TEODORO, A.K.M.2; ZANNA, R.L.D.3; MEIRA, C.S.3; GAVA, R.3; MOTOIE, G.3; PEREIRA-CHIOCCOLA, V.L.3; ALLEGRETTI, S.M.2 1.DEPARTAMENTO DE VIGILÂNCIA À SAÚDE, SMS MONTE MOR, MONTE MOR, SP, BRAZIL; 2.DEPARTAMENTO DE BIOLOGIA ANIMAL, INSTITUTO DE BIOLOGIA, UNICAMP, CAMPINAS, SP, BRAZIL; 3.CENTRO DE PARASITOLOGIA E MICOLOGIA, INSTITUTO ADOLFO LUTZ, SÃO PAULO, SP, BRAZIL.

Keyword:animal reservoir; leishmania; trypanosoma cruzi

Abstract: As part of the municipal zoonoses vigilance and control service of the Public Health Department of Monte Mor County, São Paulo state, Brazil, 42 free living wild small mammals were captured, restrained and inspected during a 36 months period, for identification of eventual natural infection by trypanosomatidae protozoa. Most of the animals were collected after casual encounter by local citizens in rural, urban or peri-urban areas, including backyard of residences.

Blood and skin samples were collected from each animal. Blood smears were prepared right after sampling and skin samples were frozen until lab processing. Direct parasite detection by light microscopy on blood smears and PCR for Leishmania sp. and Trypanosoma cruzi was conducted on the skin samples. Total captured mammal specimens per species were: Didelphis albiventris (34), Didelphis aurita (2), Coendou prehensilis (4), Gracilinamus sp. (1) and Cavia aperea (1). Leishmania sp. infection was identified in 4.76% (2/42) of the examined animals (one D. albiventris and one C. prehensilis). T. cruzi infection was found in 7.14% (3/42) of the small mammals examined through PCR (two D. aurita and one C. aperea). The samples from two Didelphis aurita with positive PCR for T. cruzi had also tripomastigote forms found on the blood smear analyses. Despite of the low prevalence of infection, wild small mammals were confirmed as natural hosts of tripanosomatidae in the studied area. Didelphis albiventris and Didelphis aurita which have synanthropic habits are a potential source of infection of Leishmania sp. and T. cruzi, and may link wild epizootic cycles to urban and peri-urban zoonotic cycles of such important zoonoses causative agents.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1292] P 912 - SAND FLIES (DIPTERA, PSYCHODIDAE) ASSOCIATED WITH NESTS OF DIDELPHIS ALBIVENTRIS (DIDELMORPHIA, DIDELPHIDAE) IN URBAN AREA CUTOLO, A.A.1; TEODORO, A.K.M.2; OVALLOS, F.G.3; ALLEGRETTI, S.M.2; GALATI, E.A.B.3

1.DEPARTAMENTO DE VIGILÂNCIA À SAÚDE, SMS MONTE MOR, MONTE MOR, SP, BRAZIL; 2.DEPARTAMENTO DE BIOLOGIA ANIMAL, INSTITUTO DE BIOLOGIA, UNICAMP, CAMPINAS, SP, BRAZIL; 3.DEPARTAMENTO DE EPIDEMIOLOGIA, FACULDADE DE SAÚDE PÚBLICA, USP, SÃO PAULO, SP, BRAZIL.

Keyword:animal reservoir; epidemiology; transmission

Abstract: Leishmaniases are zoonoses caused by tripanosomatidae protozoa of the genus Leishmania transmitted by the bite of sand flies after blood feeding on infected mammals. Although the dog is considered the main reservoir of Leishmania (L.) infantum in different endemic areas of visceral leishmaniasis in Brazil, other mammalian species have been found infected with Leishmania, including opossums of the genus Didelphis, potential animal reservoirs of the parasite. This study reports for the first time the association of sand flies with nests of opossums and their offspring in the backyard of residences located in the urban area of Monte Mor County, located in the metropolitan region of Campinas city, São Paulo state, Brazil. Eleven specimens of Evandromyia cortelezzii and one specimen of Evandromyia lenti were captured along with two nests of Didelphis albiventris. Evandromyia cortelezzi is considered a secondary

vector species on the transmission of Leishmania (V.) braziliensis and Leishmania (L.) infantum in the Neotropics. The association between this sand fly species and the synanthropic marsupial Didelphis albiventris may contribute to the introduction, establishment and maintenance of urban and peri-urban zoonotic transmission of Leishmania and should therefore be further investigated.

- RESERVOIRS [1301] P 913 - STUDY OF CUTANEOUS LEISHMANIASIS RESERVOIRS IN NORTHERN BARAAN REGION OF ISFAHAN, IRAN SABERI, S.1; HEJAZI, S.H.1; JAFARI, R.2; ARJMAND, R.1; JAVADI, A.3 1.DEPARTMENT OF PARASITOLOGY AND MYCOLOGY, SCHOOL OF MEDICINE, ISFAHAN UNIVERSITY OF MEDICAL SCIENCES, ISFAHAN, IRAN, ISLAMIC REPUBLIC OF; 2.ISFAHAN HEALTH RESEARCH STATION , NATIONAL INSTITUTE FOR HEALTH RESEARCH, TEHRAN, IRAN, ISLAMIC REPUBLIC OF; 3.DEPARTMENT OF SOCIAL SCIENCES, SCHOOL OF MEDICINE, QAZVIN UNIVERSITY OF MEDICAL SCIENCES,, QAZVIN, IRAN, ISLAMIC REPUBLIC OF.

Keyword:cutaneous leishmaniasis , reservoirs; isfahan, iran, rhombomys opimus ; meriones libycus

Abstract: Introduction: Repeated referrals of patients with cutaneous leishmaniasis to treatment centers from North Baraan especially the two villages(Timiyart and Fesaran(and also frequent local reports of the proliferation of rodents in the area, made probable spread of the disease from areas adjacent to these areas. Previously there was no study about the genus and species of regionâ&#x20AC;&#x2122;s rodents. In this study the rodent fauna of the region and also infection rate of leishmania in the hunted rodents was evaluated.

Material and Methods: The study was a descriptive cross-sectional one. Initially active colonies were identified. Using Sherman live traps the rodents were captured and based on key features family, genus and species of them were detected. Samples were taken from inflammations and clinical changes of surface body parts such as ears, snout, paws, nose and tails for direct microscopy and culture in to the N.N.N medium. Afterwards the rodents were dissected and two samples to direct smears and culture were prepared from internal visceral organs.

Results: In this study, 53 species of rodents were caught. Among them 38 rodents (71.7%) were Rhombomys opimus and 15(28.3%) was Meriones libycus.

The infection rate of leishmania in R.opimus was (18.4%) and of M.libycus was (13.3%).

Discussion: Based on the results R.opimus was reported as the principal reservoir of CL and M.libycus as the secondary host in this area.

- EPIDEMIOLOGY [1305] P 914 - MOLECULAR EPIDEMIOLOGY OF CUTANEOUS LEISHMANIASIS IN VENEZUELA RODRIGUEZ, N.M.; DE LIMA, H.; FERNANDEZ, A.; BARRIOS, M.A. UNIVERSIDAD CENTRAL DE VENEZUELA, CARACAS, VENEZUELA.

Keyword:cutaneous leishmaniasis; molecular epidemiology; reservoirs

Abstract: MOLECULAR EPIDEMIOLOGY OF CUTANEOUS LEISHMANIASIS IN VENEZUELA 1Noris Rodríguez, PhD,2 Héctor De Lima, MD; 2Alexis Fernández; 2 Miguel A. Barrios.1Instituto de Biomedicina; Facultad de Medicina; Universidad central de Venezuela.2.

Ministerio del Poder Popular para la Salud. Email: nmrodric@gmail.com In Venezuela the leishmaniasis is considered a public health problem; according with the reports from the Ministerio de Salud; between 1968 and 2012 a total of 58,402 cases of cutaneous leishmaniasis have been reported, distributed in all States of the country with a high incidence in Miranda, Lara and Merida States.(De Lima et al, 2011). However the total number of cases is unknown because the sub register is calculated in 30% due to diagnosis problems. In order to realize a better diagnosis in human cases as well as identify natural reservoirs and natural infected sand flies we have developed several molecular tools to be applied in molecular epidemiological studies in the endemic areas of the country.

The molecular tools such as molecular probes (Rodriguez et al; 1997; 2000) and species specific oligonucleotides (Rodriguez et al; 2002; 2004) have been obtained from the nuclear and the kinetoplast DNA from different New World Leishmania species together with different molecular techniques such as: schizodemes analysis, Southern blot and dot blot hybridized with species specific probes together with species specific PCR reaction has allowed to evaluated 7,350 samples from human, wild animals and sand flies .

The results obtained in diagnosis and molecular identification of the Leishmania parasites demonstrated that cutaneous leishmaniasis is mainly produced by L.(V.) braziliensis (82%) and L.(L.) mexicana (12%), L.(V.) guyanensis (1%) and other species (5%). We also achieved the identification of L.(L.hagasi producing an atypical form of cutaneous leishmaniasis in a rural area of Guarico State, Venezuela. We also identified Lu. gomezi, Lu.ovallesi and Lu. panamensis naturally infected with L.(V.) braziliensis in different endemic areas; additionally Lu. ovallesi was also found naturally infected with L.(L.) mexicana in endemic areas of Miranda state. Regarding the wild animals we found black rats (Rattus sp) naturally infected with L.(V.) guyanensis in Merida state, but Rattus rattus and Sigmodon hispidus were incriminates as the reservoirs for L.(V.)braziliensis and L.(L.)mexicana. We conclude that the molecular tools developed in our laboratory and applied in the field were very useful in order to know the epidemiological cycle of L.(V.) braziliensis and L.(L.) mexicana in endemic areas of cutaneous leishmaniasis in Venezuela, we also known the distribution of both species around the country.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1308] P 915 - LUTZOMYIA WELLCOMEI (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) IN AN ATLANTIC FOREST AREA IN THE METROPOLITAN AREA OF NATAL, RIO GRANDE DO NORTE, NORTHEASTERN BRAZIL.

PINHEIRO, M.P.G.; SILVA, J.H.T.; SIQUEIRA, L.L.D.N.; SILVA, M.M.M.; ALVES, M.L.; FREIRE, R.C.M.; XIMENES, M.F.F.M. UFRN, NATAL, RN, BRAZIL.

Keyword:lutzomyia wellcomei; lutzomyia longipalpis; atlantic forest

Abstract: Leishmaniases, which are diseases distributed worldwide, affect humans and animals in their visceral and cutaneous forms. The latter has been associated, among other sandfly species, to Lutzomyia wellcomei Fraiha, Shaw & Lainson, initially found in the Amazon region and more recently in forest areas of Northeastern Brazil. For this study, monthly captures were conducted from January to December, in an Atlantic Forest fragment and peridomestic environment in the city of Parnamirim, located in the metropolitan region of Natal, the capital of Rio Grande do Norte state. Sandflies were collected in CDC light traps, installed 1 meter above the ground between 5 pm and 7 am. They were then identified and analyzed in relation to rainfall, temperature and humidity at the collection site. Some specimens of L. wellcomei were metalized under carbon tape, visualized and photographed under a scanning electron microscope. Most species occurred in the dry season. Of the total captured in forest and peridomestic environments, around 8% corresponded to L. wellcomei (85% females and 15% males), predominantly in the forest environment, and unlike other species, after the onset of the rainy season. The most abundant species in the forest area was Lutzomyia walkeri, followed by L. wellcomei. In the forest, L. wellcomei was found along with Lutzomyia whitimani and Lutzomyia intermedia. L. wellcomei were not captured in the peridomestic environment, where Lutzomyia longipalpis was predominant, with 78% males and 22% females. Microscopic analysis of the external morphology of L. wellcomei reveals details of the male and female head, primarily the mouth parts. The present study, the first systematized investigation of this species in Rio Grande do Norte, widens knowledge on the occurrence of species suspected of transmitting Leishmania in Northeastern Brazil, in addition to instigating new research on the bioecology of sandflies in urban areas.

- EPIDEMIOLOGY [1312] P 916 - RISK FACTORS OF CHILDHOOD LEISHMANIASIS LESIONS HEALING TIME: A MULTIVARIATE MARGINAL COX REGRESSION ANALYSIS. TOUMI, A.1; BETTAIEB, J.1; MOKNI, M.2; ESSID, A.1; SALAH, A.B.1 1.DEPARTEMENT OF MEDICAL EPIDEMIOLOGY, LABORATORY OF TRANSMISSION, CONTROL APASTEUR INSTITUTE OF TUNIS, TUNIS, TUNISIA; 2.DEPARTEMENT OF DERMATOLOGY, LA RABTA HOSPITAL, TUNIS, TUNISIA.

Keyword:childhood zoonotic cutaneous leishmaniasis; lesion healing time; multivariate survival data

Abstract: Zoonotic Cutaneous Leishmaniasis (ZCL) is a seasonal and endemic disease in central and southern Tunisia, where it is caused by Leishmania (L.) major. Transmission of L. major occurs during the summer and active lesions in humans tend to emerge during autumn and winter. It affects both adults and children involving often only the skin, and may be characterized by one to dozens of lesions. Most skin lesions heal spontaneously; However, some lesions may persist for long periods or leave scars. This study attempts to elucidate the risk factors associated with ZCL lesion healing time in childhood. A prospective study was conducted in primary schools from central and southern Tunisia. Eighty three children with 215 ZCL lesions were randomly selected and followed closely for two years (April 2001- April 2003) to assess the natural evolution of their skin lesions. The Kaplan-Meier method was used for the time-to-healing analysis. Univariate and multivariate marginal Cox proportional hazard models were applied to determine risk factors for lesion healing time. Risk factors investigated included gender, age,

previous personal or family history of leishmaniasis, Leishmanin skin test response, month of lesion onset, clinical form, location, and color of the lesion. The median age of the study population at enrollment was 8 years (Interquartile range [IQR]: 7- 9). The F:M sex ratio was 1.3. The median lesion number was 1 (IQR: 1- 2), and the majority of lesions were papulonodular (95%). Lesions affected mostly the limb (84%), and less face (12%) and trunk (4%).

Survival analysis suggested that median time to heal was 18 weeks. All lesions healed within 8 months. Multivariate analysis indicated that only month of lesion onset had significant effect on healing time. Lesions which emerge early healed faster than those which appeared later, with a significant increasing trend in the adjusted odds ratio of healing time over the 4 months of lesion onset [from September (reference month) to December: 2.1 (Confident Interval [CI]95%: 1.3, 3.5); 3.2 (CI95%: 1.7, 6.0); 8.6 (CI95%: 4.7, 15.6); p < 0.001]. Large prospective studies of natural history of ZCL lesions are needed to further characterize factors that might influence the healing process dynamics.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1315] P 917 - SANDFLIES IN A PERIDOMESTIC AREA OF THE METROPOLITAN REGION OF NATAL, RIO GRANDE DO NORTE, BRAZIL. ALVES, M.L.; PINHEIRO, M.P.G.; SILVA, V.P.M.; SILVA, M.M.M.; SIQUEIRA, L.L.N.; FREIRE, R.C.M.; XIMENES, M.F.F.M. UFRN, NATAL, RN, BRAZIL.

Keyword:periurban area; sandflies; leishmaniases

Abstract: In different regions of Brazil, alterations in the environment have been changing the epidemiological profile of leishmaniases, resulting in contact with sandfly species adapted to new ecotopes and new survival conditions. The number of notified cases of visceral and cutaneous leishmaniasis cases in Rio Grande do Norte since 2009 has been similar. However, their distribution in different geographic regions in the state can be isolated or concomitant. The occurrence of leishmaniases in periurban areas indicates the evolution of a peridomestic transmission cycle, contributing to the growing incidence of human cases and deaths as well as adaptation of their vector to modified environments. This study aims at identifying and analyzing the distribution of sandfly vectors and abiotic interactions, in intradomestic and peridomestic settings, in a periurban area of the metropolitan region of Natal. Insects were captured with CDC light traps over a three-day period, at a same site in the community of Guanduba in the city of São Gonçalo do Amarante. Rainfall, which was low in the Northeast region in 2012, reached its highest level in São Gonçalo do Amarante (288 mm) in June, while temperatures averaged 28ºC throughout the year. A total of 326 sandfly specimens were collected in 2012, 236 males (73%) and 87 females (27%). Of these, 2% were captured inside residences and 98% in the back yard, with the presence of fruit trees, organic matter and the following domestic animals: fifteen chickens, one dog and one cow. Approximately two hundred people live on the street, in an area adjacent to a school, a health post and a church. The results show the concomitant presence of species involved in the transmission of etiological agents of visceral and cutaneous leishmaniasis and a consolidated peridomestic biological cycle of L. longipalpis, L. evandroi and L. whitmani. In 2012 twenty-eight cases of visceral leishmaniasis were notified in the metropolitan region of Natal. The findings of studies conducted in Rio Grande do Norte corroborate other results and discussions about the relationship between environment, proliferation of vector insects and human health. This demonstrates the paradigm of economic development along with ecological sustainability, and aims at contributing with environmental surveillance and epidemiological actions, in addition to a decrease in the environmental degradation process and leishmaniasis proliferation in urban areas.

- EPIDEMIOLOGY [1318] P 918 - ASSESSMENT OF ATTITUDES, KNOWLEDGE, PRACTICES AND EPIDEMIOLOGICAL SURVEY IN EQUADOR ENDEMIC COUNTRY OF AMERICAN TEGUMENTARY LEISHMANIASIS BORJA-CABRERA, G.P.1; HIDALGO, J.O.Q.2; SANCHEZ, A.P.C.2; CERDA, R.M.G.2; RENTERÍA, J.A.C.2; NUNES, D.E.C.2; ROBLES, J.J.T.2; CAGUANO CAZA, W.A.; YUNA, A.G.S.2; GUACHAMÍN, P.E.Y.2; IZQUIERDO, A.Y.2; TUMBAICO, J.W.T.3; CÓRDOVA, G.R.2; SIMBAIA, M.G.S.2; CAÑADAS, K.E.B.2; SURANGO, B.P.S.2; GARÓFALO MINA, K.L.4; FIERRO, M.N.A.2; VALVERDE, J.L.G.2; ARROYO, F.A.5

1.UNIVERSIDAD CENTRAL DEL ECUADOR, UNIDAD DE SALUD PÚBLICA, PROYECTO PROMETEO SENESCYT, QUITO, ECUADOR; 2.UNIVERSIDAD CENTRAL DEL ECUADOR, FACULTAD DE CIENCIAS MÉDICAS, CARRERA MEDICINA, QUITO, ECUADOR; 3.UNIVERSIDAD CENTRAL DEL ECUADOR, FACULTAD DE CIENCIAS MÉDICAS, CARRERA MEDICINA, QUITO, ECUADOR; 4.UNIVERSIDAD CENTRAL DEL ECUADOR, FACULTAD DE CIENCIAS MÉDICAS, QUITO, ECUADOR; 5.UNIVERSIDAD CENTRAL DEL ECUADOR, FACULTAD DE CIENCIAS MÉDICAS, JEFE DE LA CÁTEDRA PARASITOLOGIA, QUITO, ECUADOR.

Keyword:american tegumentary leishmaniasis; kap; equador

Abstract: The protozoa of the genus Leishmania is the causative agent of leishmaniasis. In Equador, American tegumentary leishmaniasis (ATL) is known as sarnan brava, occurs in 20 of 22 provinces with 3,000 to 4,500 new cases occurring annually and almost 3.1 to 4.5 million people live at risk of infection. Human ATL are caused by the Viannia subgenus in Equador and reservoir hosts for domestic transmission cycles are uncertain. In Pichincha province where is located the capital of the country, in the last years ATL has been expanding into northwestern tropical areas and Little is known about the epidemiology of the disease. In this study we began to investigate the epidemiological profile of human disease of 385 peoples from Nanegalito and Puerto Quito districts, through a descriptive study, using a data collection with a questionnaire including demographic, socioeconomic, epidemiological related cycle of the parasite and assessment of attitudes, knowledge and practices (KAP) against the disease. In the survey, 12,2% reported that had the disease in the pass. Assessing the demographic variables, 47,3% were male, 52,2% female sex and 85% mestizos. The socio-economic variables have shown that 53,7% have only elementary and 23,5% secondary school, only 11,4% have superior education and 53% are laborers. Respect to housing, they said that 39.8% are of cement and 38.5% of wood, 69.5% have piped water and 62.5% sewerage, 87,1% have the presence of backyard, 75,2% earthen floor, 58,4% shaded and 48,3% with moist. We identify the presence of animals, predominantly dogs (53,2%) and chickens (36,5%). The animal waste are thrown into the yard (38%), the garbage collected in plastic bags to be removed by the municipal collection (84,8%).

Between 18 to 22 h, 80% of people stay indoors and 50% use insecticides, 69% previously heard talk about leishmaniasis and only 27% know how to prevent the disease using measures as spray insecticide inside the home (13,7%), placing nets on windows and doors (20%), clean the home environment (10,3%) and not housing near vegetations (0,13%) of them. ATL in Equador is known to have a wild cycle, we are researching if ATL suffered urbanization. To confirm, we collected dogs blood samples for seroprevalence and parasite identification that are been processing to confirm domestic transmission and we have also launched several education campaigns together to Ministry of Public Health to improve knowledge, attitudes and practices on prevention.

- EPIDEMIOLOGY [1321] P 919 - EPIDEMIOLOGICAL CHANGES OF AMERICAN VISCERAL LEISHMANIASIS IN PARAGUAY IN THE LAST 10 YEARS. CANESE, A.; MACIEL, J.; ROA, L. PROGRAMA NACIONAL DE CONTROL DE LAS LEISHMANIOSIS, ASUNCIÓN, PARAGUAY.

Keyword:visceral leishmaniasis; paraguay; rk39

Abstract: American visceral leishmaniasis (AVL) has increased considerably over the past 10 years in Paraguay, corresponding this with a large endemic in canine reservoir in urban, peri-urban and rural areas. The aim of this study was to establish the epidemiological indicators of AVL in Paraguay, in the last 10 years.

We used data of confirmed cases of AVL contained in the records of the National Leishmaniasis Control of Paraguay.

In 2012 there were 76 cases of AVL. 62% of cases in 2012 came from the Central Department, 16% of Cordillera, Itapúa 7%, while Asunción and Paraguarí with 4% each. Together these four departments had 93% of all registered cases, coinciding with the more urbanized areas of Paraguay. However, there are also cases of human AVL in 8 other departments. The incidence rate per 100,000 inhabitants of the LVA was 1.14 for all of Paraguay. The lethality of the AVL was maintained between 20.8 and 7.8%, on average was 8.5%. The model case detection AVL: splenomegaly and fever, with confirmation by the rK39 serology has a sensitivity of 97%. As for direct examinations, using bone marrow puncture, which has a sensitivity of 92%? Most cases present with AVL hepatosplenomegaly, fever (not very high and intermittent, usually evening), lethargy, weight loss, anemia, leukopenia, neutropenia and thrombocytopenia.

The AVL epidemiological reality is different from 10 years ago. Have significantly increased the number of cases and has also undergone a geographic expansion, currently covering 13 of the 18 departments of Paraguay.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1328] P 920 - EFFECTS OF THE DEFORESTATION AND HUMAN POPULATION DENSITY ON THE DIVERSITY AND ABUNDANCE OF PHLEBOTOMINE SAND FLIES (DIPTERA: PSYCHODIDAE) IN A RURAL SETTLEMENT ON BRAZILIAN AMAZON PESSOA, F.A.C.1; RAMOS, W.R.2; JULIÃO, G.R.3; LUZ, S.L.B.1; DE MEDEIROS, J.F.3 1.LEÔNIDAS & MARIA DEANE, FIOCRUZ AMAZÔNIA, MANAUS, AM, BRAZIL; 2.INSTITUTO NACIONAL DE PESQUISA DA AMAZÔNIA, MANAUS, AM, BRAZIL; 3.FIOCRUZ RONDÔNIA, PORTO VELHO, RO, BRAZIL.

Keyword:amazonia; environmental impacts; cutaneous leishmaniasis

Abstract: The sand fly are vectors of many parasites, including flagellates which cause leishmaniasis.

These insects can be found predominantly in forests, however, the impact of deforestation and colonization of these areas shown that the adaptation of some species in these environments. We realized a study of the sand flies fauna from Rio Pardo settlement, Presidente Figueiredo Municipality, Brazil. We observed the abundance and diversity of sand flies in four categories with differents degrees of deforestation and human population density: G1. High human population density and High deforestation; G2. Low human population density and high deforestation; G3. High human population density and low deforestation; G4. Low human population density and low deforestation. In each category six collections were done in the following environments - forest, edge of forest, shrub, peridomicile and orchard with light traps; overnight in June, July, August 2009 and September, October, November 2010. A total of 3.073 specimens were collected, 1.163 females (37,8%) and 1.910 males (62,2%), and identified in 13 genera and 53 species. The most abundant genera were Nyssomyia and Psychodopygus. Some

species captured five are incriminated as vectors of Leishmania: Nyssomyia antunesi (1.025 indivíduals), N. umbratilis (448), Trichophoromyia ubiquitalis (164), Psychodopygus amazonensis (82) e Bichromomyia flaviscutellata (76), found in forest and peridomicile. The largest abundances of species were observed in categories G1 and G3, both with high human population density; althought these differences were not statistically significant when compared with others; the Shannon-Wienner Indices shown the diversities in categories G2 and G3.

Among these environments, the largest sandflies abundance was observed in the peridomicile and forest environments. Main conclusions: The results may suggest that even in area with high degree of deforestation and high human population, the abundance and diversity of sand flies still maintain in high numbers. Probably, human population densities produce favorable locations for maintenance life cycle of several species, providing potential site for breending, resting, reproduction and availability of sources meal.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1329] P 921 - STUDY OF RHIPICEPHALUS SANGUINEUS (ACARI: IXODIDAE) AS POTENTIAL VECTOR OF CANINE VISCERAL LEISHMANIASIS IN THE DISTRICT FEDERAL, BRAZIL. SILVA, V.M.1; GONÇALVES, R.G.1; ARAUJO, N.N.S.L.1; CUPOLILLO, E.2; ROMERO, G.A.S.1 1.UNIVERSIDADE DE BRASÍLIA, BRASÍLIA, DF, BRAZIL; 2.FUNDAÇÃO OSWALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:leishmania infantum; rhipicephalus sanguineus; visceral leishmaniasis

Abstract: Alternative forms for the transmission of canine visceral leishmaniasis (CVL) have been investigated and sting or ingestion of infected ticks could have epidemiological relevance. The objective of this study was to detect the presence of Leishmania spp. in R. sanguineus collected from dogs naturally infected in the District Federal. Ticks were obtained from a sample consisting of 48 dogs with suspected or confirmed diagnosis of CVL, through tests. Venous blood was collected from 48 dogs for serologic evaluation of infection by Leishmania spp..

After identification of specimens of R. sanguineus ticks were dissected for DNA extraction and subsequent amplification of 120pb of the conserved region of kDNA and 234pb of the hsp70 gene of Leishmania spp. by polymerase chain reaction (PCR). Ticks sample were also cultured in modified NNN culture medium. The amplified products of kDNA were further subjected to digestion with restriction endonucleases Hae III and BstUI and gene sequencing to identify the parasite species. The positivity of serological techniques applied to canine blood was 66.6% by immunoenzymatic assay. Among positive samples, 81.2% were also positive by Indirect Immunofluorescence Assay and 56.2% by a dual path platform immunochromatographic rapid test, then, 70.8% of dogs had at least one of the three tests with positive results. Isolation of Leishmania spp. was successfull in five pools of salivary glands and intestines of ticks collected from four dogs. Sequencing kDNA amplified by PCR from these samples revealed homology with sequences of Leishmania spp. The amplified products of kDNA in a blood sample from one of the dogs showed sequence homology to L. braziliensis, however, the amplified material from ticks collected from this dog had homology with L. infantum sequence. The isolate analyzed by mulilocus enzyme electrophoresis was identified as L. infantum. The Leishmania infection in dogs where we collected the pools of ticks which had successful parasite isolation in culture was confirmed by PCR. The smear of the pool of male ticks collected from one dog was the only positive for flagellated trypanosomatidae whose identification was not possible by morphometry. Our results confirm the presence of viable L. Infantum in R. sanguineus feeding naturally in dogs naturally infected and the possibility of isolating L. infantum in culture medium from ticks. Financial support: CAPES - Basic Parasitology (032/2010).

- RESERVOIRS [1330] P 922 - GENETIC HOMOLOGY OF LEISHMANIA MAJOR STRAINS ISOLATED IN TUNISIA FROM TWO RODENTS SPP, REVEALED BY MLMT GHAWAR, W.1; ATTIA, H.2; BETTAIEB, J.1; YAZIDI, R.1; LAOUINI, D.2; SALAH, A.B.1 1.DEPARTEMENT OF MEDICAL EPIDEMIOLOGY, LR11IPT02, PASTEUR INSTITUTE ., TUNIS, TUNISIA; 2.LABORATORY OF TRANSMISSION, CONTROL AND IMMUNOBIOLOGY OF INFECTIONS (LR11IPT02),. PASTEUR INSTITUTE, TUNIS, TUNISIA.

Keyword:rodents; leishmania major; mlmt

Abstract: Zoonotic cutaneous leishmaniasis (ZCL) caused by Leishmania (L.) major parasites represents a major health problem with a large spectre of clinical manifestations. Psammomys (P.) obesus and Meriones (M.) shawi represent the most important host reservoirs of these parasites in Tunisia. We already reported that infection prevalence is different between these two rodents .

We aimed in this work to evaluate the impact of parasite genetic background on this difference.

Using the multilocus microsatellites typing (MLMT), we analyzed the genetic diversity among strains isolated from (i) P. obesus (n = 31), (ii) M. shawi (n = 8) and (iii) Mustela nivalis (n = 1), captured in Sidi Bouzid, an endemic region for ZCL located in the Center of Tunisia. Ten highly polymorphic microsatellites markers were used to study the genetic structure and microheterogeneity of these strains.All markers showed no polymorphism among the analyzed strains regardless of their host-reservoir origins.

Absence of genetic variability between strains isolated from different reservoirs can be explained by similarity of geographic and ecological parameters within the study area.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1331] P 923 - HEMATOPHAGY BEHAVIOR OF LABORATORY REARED LUTZOMYIA MIGONEI (FRANÇA) (DIPTERA: PSYCHODIDAE) IN RELATION TO THREE MAMMALIAN BLOOD SOURCES PESSOA, F.A.C.; BRUCE, K.K.M.; MARIALVA, E.F.; RÍOS-VELÁSQUEZ, C.M. CENTRO DE PESQUISA LEÔNIDAS & MARIA DEANE, FIOCRUZ AMAZÔNIA, RUA TEREZINA, 476, ADRIANÓPOLIS, 690570, MANAUS, AM, BRAZIL.

Keyword:colony; fertility; egg production

Abstract: Lutzomyia migonei (França, 1920) is widely distributed in South America. This is important vector of American Cutaneous Leismaniasis in Venezuela and Brazil. The establishment of colonies of sand flies in the laboratory is essential to study of its biology and its interaction with Leishmania. A few laboratory studies addressing behavior hematophagy sand flies using different sources such as blood meal, fecundity, fertility, and the volume of food. Thus, this study aims to verify the fertility and fecundity of females of lab reared L. migonei resource using blood from three different hosts: Mesocricetus auratus Waterhouse (hamster), Mus musculus Linnaeus (mouse) and Homo sapiens Linnaeus (man). The L. migonei used in the experiments, were from a colony initiated in 2010 (24th generation) originally from Ceará State, Brazil, which is maintained at the Laboratory of Entomology, of the Institute Leonidas & Maria Deane.

We separated 70 engorged females fed from the different blood meal sources and placed in individual oviposition tubes. After the females laid eggs and died, the eggs laid were counted and then the female was dissected and counted ing the number of mature oocytes retained in the ovarioles. We obtained eggs from 2.674 hamster blood source, 1.578 from human and 1.487

from mouse; and respectively to fecundity 2.888, 1.678 and 1.657. Descriptive statistical analyzes were performed, and the comparison of fertility and fecundity by Kruskal-Wallis (p <0.01) showed significant differences between the blood meal sources and to fertility and fecundity of L. migonei. The results show that hamster blood is the best tested supply of blood in relation to fertility and fecundity to L. migonei, and mouse blood had lowest results.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1332] P 924 - WING GEOMETRIC MORPHOMETRICS OF THREE CRYPTIC FEMALE LUTZOMYIA (PSATHYROMYIA) SPECIES FROM BRAZILIAN AMAZON RÍOS-VELÁSQUEZ, C.M.1; DE OLIVEIRA, A.F.J.2; MARIALVA, E.F.1; BARBOSA, J.C.1; PESSOA, F.A.C.1

1.ILMD FIOCRUZ AM, MANAUS, AM, BRAZIL; 2.UNIVERSIDADE FEDERAL DO AMAZONAS, MANAUS, AM, BRAZIL.

Keyword:psathyromyia; amazon; taxonomy

Abstract: The females of Lutzomyia (Psathyromyia) dendrophyla, L. (P.) scaffi and L. (P.) shannoni are difficult to identify because of their morphological similarity. These species have sympatric distribution and are considered as vectors or suspected vectors of parasites such as Leishmania.

The correct identification of sand fly species is important component for control and surveillance strategies. The aim of this work is to differentiate L. dendrophyla, L. scaffi and L. shannoni by geometric morphometrics methods. We used 23 specimens of lab reared females of L. dendrophyla, 23 of L. scaffi and 20 of L. shannoni. Field collections were done in Manaus Municipality, Amazonas, Brazil. Right wings of the specimens were slide mounted and photographed. Eight landmarks were identified and transformed in coordinate matrix using COOWin program. We used centroid size, to analyze wing size variation. Generalized Procrustes Analysis (GPA), superimposition algorithm, was used to analyze shape variables by MOG program. Data from GPA were used to compare shape variables across groups by Manova. A factor map of the first two canonical factors was used to show the main results (95% confidence interval). Anova and Manova tests were computed by Past program 1.57. We observed a significant difference in the wing size between the three studied species (H:8,439, p<0,01). L. shannoni showed the higher wing size, while L. scaffi was smaller. The difference was significant between those 2 species, but not in L. dendrophyla. Factor map showed real difference between the configurations of the three species, however some intersection was observed between L. scaffi and L. dendrophyla. ACPs calculated with wing conformation showed well grouped species. The first two principal components explained 51,04% and 16,16% of wing shape variation, respectively. Geometric morphometrics allowed to differentiate the three sandfly species. L. dendrophyla was not significantly different of the other two species, but the factorial map clearly shows the differentiation of the three groups.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1333] P 925 - THE GENITAL ATRIUM OF THREE CRYPTIC LUTZOMYIA SPECIES OF PHLEBOTOMINE SAND FLIES (DIPTERA, PSYCHODIDAE) OF SUBGENUS PSATHYROMYIA FROM CENTRAL AMAZON DE OLIVEIRA, A.J.F.1; BARBOSA, J.C.O.2; MARIALVA, E.F.2; PESSOA, F.A.C.2 1.UNIVERSIDADE FEDERAL DO AMAZONAS, AV. GENERAL RODRIGO OCTÁVIO JORDÃO RAMOS, 3000,, MANAUS, AM, BRAZIL; 2.CENTRO DE PESQUISA LEÔNIDAS & MARIA DEANE,, MANAUS, AM, BRAZIL.

Keyword:morphology; taxonomy; amazonia

Abstract: The taxonomic significance of the armature in genital atrium of sand flies were first examined

by Madulo-Leblond et al.(1991). Then, just very few studies about taxonomic importance of atrium had been done. species. The atrium is a chitinous structure shaped like a wish-bone, with a thin membrane bearing an armature with small spines between the arms of the furca. It is located towards the entrance of the oviposition duct and the atrium membrane connects to the opening of the spermathecal duct or ducts. The aim of this study was to separate criptic Psathyromia female species of the subgenus Psathyromyia, using the structures of genital atrium as a taxonomic character. Lab reared females of Lutzomyia dendrophyla, L. scaffi and L. shannoni were obtained in the entomological laboratory of Institute Leônidas e Maria Deane – ILMD/FIOCRUZ. The procedures followed to isolate the genital atrium and spermathecae closely followed those described by Killick-Kendrick et al.(1994), with modifications. the last tergal three segments of the abdomen were separated and placed in 10% KOH for a period of 24 hour. The samples were washed in acetic acid, water and placed in glycerin for dissection on a stereoscopic microscope. The segments were then separated, as in the usual procedure for isolating the spermathecae. Fine needles were then used to break open this cylinder ventrally and so expose the atrium and then slide mounted and photographed and measured the length of the furca, width of inner part of the atrium, and the shape of the spimes. Statistical analyses were done by PAST program. Nine slide mounted atrium of each studied species were obtained. The arrangement of the spines varied for each studied species: To L. dendrophyla, the spines were larges and formed one to six spine combs forming rows in the mebrane. The L. scaffi atrium armature has three to six spines combs forming a horizontal stripe in the middle of the atrium membrane. L. shannoni has combs of tyhree to six short spines in the middle of the mebrane and large individuals spines in the lateral borders of the aritum membrane. We found a significant size difference between the lengh, width and internal membrane width of the atria in L. dendrophyla and L. scaffi (p=0,0003), (p=0,0004) and (p=0,008)and L. scaffi and L. shannoni (p=0,006), (p=0,000) and (p=0,0016); None signifant difrences were found between the size of the atria structures to L. dendrophyla and L. shannoni . We concluded that the genital atrium is a interesting and useful tool to distinguish in some casdes, criptical female species of Psathyromyia subgenus.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1334] P 926 - DESCRIPTION OF THE LARVAE OF THREE SPECIES OF LUTZOMYIA (PSATHYROMYIA) (DIPTERA: PSYCHODIAE) FROM BRAZILIAN AMAZON MARIALVA, E.F.1; OLIVEIRA, A.F.J.2; BARBOSA, J.C.O.1; PESSOA, F.A.C.1

1.CENTRO DE PESQUISA LEÔNIDAS & MARIA DEANE, FIOCRUZ AMAZÔNIA, RUA TEREZINA, 476, ADRIANÓPOLIS, 690570, MANAUS, AM, BRAZIL; 2.UNIVERSIDADE FEDERAL DO AMAZONAS, MANAUS, AM, BRAZIL.

Keyword:taxonomy; morphology; vectors of leishmaniasis

Abstract: Phlebotomine sand flies (Diptera:Psychodidae) have medical and veterinary importance. As with many insect groups, the morphological characters used to distinguish sand fly species are those of the adults, the immature stages of most species being unknown and/or undescribed. The larval stages of less than 70 species of the 400 new world sand flies have been described (Young & Duncan 1994). The objective of this work was to describe the mature larvae of 03 species of sand flies, belong to Psathyromyia subgenus. The larvae of Lutzomyia dendrophyla, L. scaffi e L. shannoni were obtained from laboratory colonies following the methods of Killick-Kendrick & Killick-Kendrick (1991) and reared from adults originally collected in Presidente Figueiredo, Amazonas State, Brazil. Four slides of 4th instar of each species were slide-mounted in Berlese’s fluid. Mouthparts of other three larvae were dissected and then placed on 10% KOH for 02 days, then slide-mounted. Measurements of the body bristles and morphological

structures as mouthparts, setae, spiracles were made under light microscopy and photographed.

Morphology and chaetotaxy of the head and body were observed following Ward (1976).

General appearance of the larva: the larva is caterpillar-like, with a well sclerotized hypognathous non-retractile head. Antennal tubercle long, conspicuously aboral, longer than antenna. Antenna slender, first segment longer than second, the latter directed backward at slight angle from first segment. Integument covered with numerous spicules, arranged into imbricated design posteriorly. Thorax and abdomen flattened dorso-ventrally, rather short and broad.

Integument covered with spicules, which on summits of dorsal swellings grade into rounded tubercles. Coloration: Head brown, thorax and abdomen light than, with dorsal brown path on abdominal segment VIII. Caudal setae dark. General size body around 5 mm of length The L. scaffi larva was the largest length with 5,7 mm and L. shannoni the smallest with 4,7 mm. The main chaetotaxy differences between the species, with significant different sizes were the dorsolateral setae, anterior ventrolateral and internal caudal setae.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1337] P 927 - PRELIMINARY LIFE CYCLE ASPECTS OF THREE LAB REARED LUTZOMYIA (PSATHYROMYIA) AMAZONIAN SPECIES BARBOSA, J.C.O.1; OLIVEIRA, A.F.J.2; MARIALVA, E.F.1; PESSOA, F.A.C.1 1.CENTRO DE PESQUISA LEÔNIDAS & MARIA DEANE, FIOCRUZ AMAZÔNIA, RUA TEREZINA, 476, ADRIANÓPOLIS, 690570, MANAUS, AM, BRAZIL; 2.UNIVERSIDADE FEDERAL DO AMAZONAS, MANAUS, AM, BRAZIL.

Keyword:colony; criptic species; amazonia

Abstract: Laboratory colonization of sand flies is invaluable for the progress in research on insect biology, physiology, behavior, vector competence, taxonomic studies etc. Sand flies of the genus Lutzomyia subgenus Psathyromyia have public health importance because they are suspected vectors of arboviruses and some trypanosomatid protozoans such Leishmania. Some females of Psathyromyia species are identified with difficulty because they are morphologically similar species e.g. L. shannoni and L. scaffi. Due to the necessity to obtain enough material to taxonomic studies and information of basic biology, we reared in laboratory conditions those species during the three first generations. This study aimed to describe the basic aspects of the life cycle of these species in the laboratory. Sand flies were caught in the field and the in pregnant or engorged females were separated and placed in individual oviposition tubes.

Isofemale broods are placed to individual pots, with a thin layer of plaster of Paris and kept in BOD incubator at 26.5ºC and 90% of humidity. The colonization method was based on KillickKendrick & Killick-Kendrick (1991) The larval diet according to Young et al. (1981), adding fish larvae, with 55% crude protein. Each pot was inspected daily to check the eggs hatch, larvae number, and monitoring of larval instars changes. The species identification was made after the emergence of adult males at the end of the cycle. The average time span (in days) to all life cycle, since the oviposition to adult emergence, to L. shannoni and L. scaffi , respectively were 46,4 (N= 24) and 45 (N= 72). Eggs 10,5 (N= 99) and 8,6 (N=173) 1st instar 7,7 (N=41)and 09 (N=116), 2nd instar 8 (N=37) and 6,3 (N=98) , 3rd 7 (N=35) and 6,8 (N=94), 4th 10,5 (N=33) and 9,8 (N=86), pupa 9,6 (N=27) and 10,5 (N=80). The reared females fed avidly on hamster and human as early as 2 days after emergence. These preliminary observations have shown that L. shannoni and L. scaffi from Amazon basin can be reared in laboratory.

- EPIDEMIOLOGY [1339] P 928 - EPIDEMIOLOGICAL ASPECTS OF CANINE VISCERAL LEISHMANIASIS IN AN ENDEMIC URBAN AREA IN MATO GROSSO DO SUL STATE, BRAZIL CUTOLO, A.A.1; MOREIRA, M.A.B.2; NOGUEIRA, F.S.3; RUI LUVIZOTTO, M.C.4; CARVALHO, F.G.5; MENZ, I.6

1.DEPTO DE VIGILÂNCIA À SAÚDE, SMS MONTE MOR, MONTE MOR, SP, BRAZIL; 2.HOSPITAL VETERINARIO, UNIVERSIDADE ANHEMBI MORUMBI, SÃO PAULO, SP, BRAZIL; 3.FUNDAÇÃO EDUCACIONAL DE ANDRADINA, FEA, ANDRADINA, SP, BRAZIL; 4.FACULDADE DE MEDICINA VETERINARIA, UNESP, ARAÇATUBA, SP, BRAZIL; 5.CENTRO DE CONTROLE DE ZOONOSES, CCZ, CAMPO GRANDE, MS, BRAZIL; 6.DEPARTAMENTO DE BIOLOGIA ANIMAL, INSTITUTO DE BIOLOGIA, UNICAMP, CAMPINAS, SP, BRAZIL.

Keyword:: incidence; incubation time; diagnostics

Abstract: Visceral leishmaniasis (VL) incidences in human and canine hosts are increasing and expanding in Brazil. In order to perform an adequate vigilance of this disease, the knowledge of epidemiological parameters like prevalence, incidence, incubation period and potential of transmissibility in the dog, the main reservoir for the VL etiologic agent is mandatory. This way, a longitudinal study in a VL endemic area located in Campo Grande, Mato Grosso do Sul state, Brazil, was performed during a two years period. Hundred and five dogs were evaluated through clinical examination, serology (ELISA L. major-like, FML and S7) and direct diagnostic techniques (bone marrow and lymph node smears and PCR and immunohistochemistry in skin biopsy) on dogs. A seroprevalence of 30.40% (145/477) at ELISA L. major-like and 26.28% (97/369) at ELISA FML was obtained at the pre-initial phase. Clinical disease prevalence was 7.13% (62/870), and 24.53% (117/477) dogs were seropositive at ELISA L. major-like, but asymptomatic, showing the high amount of infected healthy carriers. At the end of the 24 months period no significant association was found between canine cases of visceral leishmaniasis (VL) with environmental variables evaluated at Day Zero like: previous registration of human or canine VL on the study house or neighborhood, other domestic animal species at the backyard, intra or peridomicile, aggressive behavior, age or sex of the study dog.

Incidence of infection varied from 11.76% (8/68), 11.76% (8/68) e 18.18% (12/66) respectively between Day -30 and Zero, Day Zero and Month 12 using serology and Day Zero and Month 12 for direct diagnostic methods. Natural and induced mortality through euthanasia of sick/infected dogs was 28.57% (30/105). The mean disease incubation period was 6.46 months. Some of the infected animals with positive diagnosis at the beginning of the study had no other positive results after 12 months, indicating a possible cure or an absence of infection establishment, while others had positive diagnosis at the beginning and at month 12 with asymptomatic condition for more than 24 months. A low Leishmania amastigote prevalence at healthy skin was obtained and positive results were observed only in symptomatic animals. Absence of parasites in skin of asymptomatic carriers may indicate they have no important role on the transmission of Leishmania to competent vectors.

- DIAGNOSIS – EXPERIMENTAL AND CLINICAL [1342] P 929 - VALIDATION OF A STRATEGY OF COMBINED TESTING FOR SEROLOGICAL DIAGNOSIS OF CANINE VISCERAL LEISHMANIASIS IN BRAZIL ARRUDA, M.M.1; FIGUEIREDO, F.B.2; CARDOSO, F.A.3; BARBOSA, J.R.3; NORONHA, E.F.1; ROMERO, G.A.S.1 1.NUCLEO DE MEDICINA TROPICAL - UNB, BRASÍLIA, DF, BRAZIL; 2.INSTITUTO DE PESQUISA CLINICA EVANDRO CHAGAS- FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.FUNDAÇÃO EZEQUIEL DIAS, BELO HORIZONTE, MG, BRAZIL.

Keyword:sensitivity and specificity; canine visceral leishmaniasis; diagnosis

Abstract: Current recommendations by WHO for controlling zoonotic visceral leishmaniasis due to Leishmania infantum include serological canine surveys to define the distribution and frequency of the infection in dogs. However, the lack of accuracy of diagnostic tests may cause the permanency of infected dogs in the community or the unnecessary euthanasia of dogs with false positive results. The combination of tests could optimize accuracy of serological diagnosis of canine visceral leishmaniasis (CVL). The objective of this study was to estimate the accuracy of

ELISA and IFA (Bio-Manguinhos 速, Rio de Janeiro, Brazil) which are currently used in the routine of the Visceral Leishmaniasis Brazilian Control Program applied in parallel or sequential testing. The assays were conducted at the Foundation Ezequiel Dias (FUNED), Belo Horizonte Minas Gerais, the National Reference Laboratory for serological diagnosis of CVL. We used a panel of 1,425 dogs sera collected between 2008 and 2010. The animals were randomly selected and classified as infected or not infected using the combination of at least one positive result in the following tests on a skin sample as the gold standard: parasitological examination in histopathological specimens stained with H&E, immunohistochemical examination or parasite isolation in culture. Serological tests were applied according to the manufacturer's instructions and the reading was performed by two reviewers independently and blinded to the results of the gold standard. Of the 1425 dogs, 98 (6.9%) were classified as positive and 1327 (93.1%) as negative by the gold standard. The sensitivity was 91.8% (95%CI 86,33 to 97,27) and 90.8% (95%CI 84,15 to 97,48) for ELISA and IFI, respectively. The specificity was 83.4% (95%CI 81,93 to 85,85) and 53.4% (95%CI 51,59 to 55,12) for ELISA and IFI, respectively. When used in parallel, the sensitivity increased to 99.2% but the specificity decreased to 44.8%. When used in series (ELISA followed by IFA), sensitivity decreased to 83.3% and specificity increased to 92.5%. The positive predictive value of the tests in series was four times higher (44% versus 11%) than the tests applied in parallel and the negative predictive value remained high (99%)for both approaches for the actual prevalence of infection observed in the sample. Based on these results we suggest that the use of ELISA and IFI tests in a sequential manner would be the most suitable for the diagnosis of canine infection.

- EPIDEMIOLOGY [1343] P 930 - THE VETERINARY MEDICAL OFFICER OF PMERJ AND THEIR ROLE IN SURVEILLANCE AND CONTROL OF CANINE VISCERAL LEISHMANIASIS NAURATH, P.E. PMERJ, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:leishmania; military kennels; epidemiology

Abstract: This article had as objective to investigate the paper of the veterinary medical officer of PMERJ (Military Police of Rio de Janeiro City) in the scope of monitoring epidemiology and control of Canine Visceral Leishmaniosis in the canine kennel of Military Police of Rio de Janeiro City, comparing it with military kennels of Federation.and on the State of Rio De Janeiro. Survey from questionnaires sent by electronics was carried through the Veterinary medical officer of PMERJ, military kennels of the State of Rio De Janeiro, and also of other States of the Federacy aiming to evaluate the degree of clarification of the illness and its risks for the Veterinary medical officers , the half used laboratorial diagnosis more for diagnosis, as well as if the accomplishment of prophylactic procedure it is a routine and which more used for same the Twenty nine questionnaires were answered and 82,76% have already mett with animal zoonoses. Of these ones, it was questioned if they have attended any animal suspected or positive for Visceral Leishmaniasis,and 51.75% answered yes About these same respondents, the laboratory diagnosis was made only by serology (IFA / ELISA) in 33.34% of the cases, the serology IFA / Elisa cytology and biopsy of lymph node / bone marrow in 20% of cases, serology IFA / Elisa together with cytology lymph node biopsy with PCR in 20% of cases; SorologiaRIFI / Elisa and Immunohistochemistry-got in 13.33% of the cases, PCR was 16.66% of the cases, SorologiaRIFI / Elisa biopsy and lymph node / bone marrow -got a percentage of 16.66% of cases. Asked respondents who reported seeing positive animal if the animal in question was sacrificed, 73.33% spoke yes. Regarding the question of whether prophylactic procedure is performed for Visceral Leishmaniosis in your corporation, of the 29 respondents,

65.52% are to disease prophylaxis.A relevance of this work consists of the necessity of implantation of a new project of prophylactic measures of monitoring epidemiologist against the Canine Visceral Leishmaniosis in the kennel of the PMERJ, adequate the new demands of cases in the State of Rio De Janeiro city and the importance epidemiologist of this illness, being real the risk of exposition and dog contamination in PMERJ kennel, as well as secondary contamination of military policemen as treators, conductors, adestradores, to assist of services veterinarians, veterinarians and administrative staff of the unit..

- RESERVOIRS [1349] P 931 - ASSOCIATION OF PRESENCE OF LEISHMANIA INFANTUM AMASTIGOTES IN THE SKIN OF DOGS NATURALLY INFECTED BY L. INFANTUM WITH THE CLINICAL FEATURES AND LABORATORY CHANGES. RIBEIRO, V.M.1; BAHIA, E.M.2; TELES, P.P.A.1 1.ESCOLA DE MEDICINA VETERINARIA PUC MINAS, BETIM, MG, BRAZIL; 2.CLINICA VETERINARIA SANTO AGOSTINHO, BELO HORIZONTE, MG, BRAZIL.

Keyword:canine leishmaniosis; clinical features; infectivity

Abstract: The observation of amastigotes forms on the skin of dogs, through the method of immunohistochemistry (IHC) (Tafuri et al., 2004), has been presented as an indicator to the vector Lutzomyia longipalpis infectivity, having a strong correlation with the xenodiagnosis (Amorim et al ., 2011). In this study, 38 dogs were positive to amastigotes forms, using IHC, and the important clinical alterations were observed. Analyzing the clinical standard, 27/38 (71%) were symptomatic and 11/38 (29%) asymptomatic. The common clinical signs observed were lymphadenomegaly 24/27 (89%), dermatitis 21/27 (78%), alopecia 18/27 (67%), weight loss 10/27 (37%), onychogryphosis 3/27 (11%), cutaneous ulcer and otitis 1/27 (4%). The blood count performed in 36 dogs found that 26/36 (72%) had under 37% hematocrit and 10/36 (38%) within normal values. Leukopenia was observed in 10/36 (38%) and the remaining 26/36 (72%) had the leukocyte count within normal standards. When the leukocytes were analyzed, it was possible to observed that the lymphocytes, monocytes and eosinophils cells were the major influence to the leukopenia, cause 11/36 (31%) of the dogs presented lymphopenia, 11/36 (31%) eosinopenia ainfecnd 33/36 ( 92%) monocytopenia. The urea and creatinine concentration were elevated in 7/36 (19%) and 4/36 (11%), respectively. The Albumin/Globulins (A/G) was below 0.6 in 24/36 (67%) due to hypoalbuminemia in 20/36 (56%), and hypergammaglobulinemia in 26/36 (72%). The antibodies dilution by indirect immunofluorescence (IFI) was correlated with animals that were positive for amastigotes forms on the skin as dilutions: equal or greater than 1:5.120, 14/33 (42%); of 1:640 to 1:2560, 12/33 (36%) and 1:80 to 1:320, 9/33 (27%).

Conclusion: Clinical signs, anemia, hyperglobulinemia and inversion of the fraction A/G to values lower than 0.6 and higher titers in IFI are the indicators of infectivity for L. longipalpis in dogs naturally infected by L. infantum.

- RESERVOIRS [1351] P 932 - CORRELATION BETWEEN THE PRESENCE OF AMASTIGOTES FORMS OF LEISHMANIA INFANTUM IN DOGS SKIN NATURALLY INFECTED AND FACTORS AS BREED, GENDER AND AGE. RIBEIRO, V.M.1; TELES, P.P.A.1; BAHIA, E.M.2

1.ESCOLA DE MEDICINA VETERINÁRIA PUC MINAS, BETIM, MG, BRAZIL; 2.CLINICA VETERINÁRIA SANTO AGOSTINHO, BELO HORIZONTE, MG, BRAZIL.

Keyword:canine leishmaniosis; infectivity; canine breeds

Abstract: From January 2008 to July 2012, 473 dogs from 49 different breeds naturally infected by Leishmania infantum were examined. Out of this total, it was found that 38 presented amastigotes forms on the skin on the apex of the inner ear, where the major parasite concentration is considered to locate (Xavier at al., 2006). The method used for the detection of the parasites was the immunohistochemistry (IHQ), which has a strong correlation with xenodiagnoses, therefore it is considered a good predictor of infectivity and evaluator of Lutzomyia longipalpis in dogs naturally infected with L. infantum (Amorim et al., 2011). In 15 of the 49 analyzed breeds, the presence of amastigotes forms on the skin was detected. Although the percentage of infectivity found for each breed was 6% (2/32) in Daschund, 8% (7/84) Mongrel, 10% (1/10) Shitzu, 11% (7/66) Poodle, 13% (2/16) Boxer, 13% (1/8) Schnauzer, 17% (1/6) French Bulldog, 19% (3/16) Labrador Retriever, 20% (1/5) Great Dane, 20% (1/5) Brazilian Fox, 20% (1/5) Pug, 22% (2/9) Lhasa-Apso, 25% (1/4) Brazilian Fila, 33% (3/9) English Bulldog and 45% (5/11) Basset Hound. The number of male animals with positive skin (39%) was lower than the number of female animals (61%) in a total of 242 males and 231 females. In age distribution were found 15/98 (15%) positive dogs between 0 and 24 months, 14/188 (7%) between 25 to 72 months and 9/187 (5%) over 72 months. Considering only the positive animals, 39% were between 0 and 24 months, 37% between 25 and 72 months and 24% over 72 months.

Conclusions: The results allow us to conclude that dogs of various breeds can be infectious.

Although the number of dogs per breed is different, the highest positive percentage was seen in the Basset Hound, English Bulldog and Brazilian Fila breeds. Females seem to be more infectious which could be related to the hormonal interference. Also, young and adult animals are more likely to be infectious compared to the elderly.

- EPIDEMIOLOGY [1353] P 933 - ENTOMOLOGICAL MONITORING OF THE PROGRAMME OF VISCERAL LEISHMANIASIS IN MACEIÓ, ALAGOAS STATE, BRAZIL: INDICATORS AND ENVIRONMENTAL RISK FACTORS ROCHA DOS SANTOS, C.F.1; LIMA, M.C.1; ROCHA NETO, G.B.1; ZWETSCH, A.2; VILELA, M.L.2; SENA, J.M.3

1.SECRETARIA MUNICIPAL DE SAÚDE DE MACEIÓ, MACEIÓ, AL, BRAZIL; 2.LABORATÓRIO DE TRANSMISSORES DE LEISHMANIOSES/ INSTITUTO OSWALDO CRUZ/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.SECRETARIA DE VIGIL}NCIA EM SAÚDE- MINISTÉRIO DA SAÚDE, BRASÍLIA, DF, BRAZIL.

Keyword:maceió; flebótomos; lutzomyia

Abstract: The city of Maceió is ranked, according to the methodology of the Ministry of Health as sporadic transmission to American Visceral Leishmaniasis (AVL). The occurrence of cases follows the process of human occupation, located in discontinuous areas and sparse, with half of the territory is comprised of rural areas, integrating even this scenario, a park and an area of environmental preservation. Entomological surveillance actions were triggered initially aimed to determine the distribution of the vector, Lutzomyia (Lutzomyia) longipalpis through entomological survey and monitoring, in order to know the seasonality. Monthly collections were made with the help of CDC light traps in the period from July 2004 to December 2007, and selected areas for implementation of entomological monitoring from identifying the points of greatest abundance of sand flies in the entomological survey, correlated to the presence of

human and canine cases of the disease. The traps were installed from 18h and 07h drawn around the next day for three consecutive nights, totaling 20 traps indoors and peridodomestic sites. We collected 9652 sand flies and recorded nine species, especially L. (Nyssomyia) whitmani (62%), L. (Lutzomyia) longipalpis (26%) and L. migonei (10%), all other species amounted to 2%. The locations of the Povoado Canto do Farias and Sitio São Gonçalo were those which had the highest abundance vector. Entomological indicators showed variation according to the presence of domestic animal shelters around the houses, and also regarding the conditions of residence and can be seen a decrease in vector abundance inside the house after improvements made. The results showed that there was a relationship between sand fly density and housing conditions and environmental factors, especially the presence of domestic animals (chickens and pigs).

Environmental characteristics and lack of adequate infrastructure in communities, and how precarious proximity with the forest, presence of pets around the houses, garbage accumulation, favored the maintenance of sand fly vector breeding sites, increasing the risk of disease transmission. The entomological data found allies to environmental risk factors identified in the area suggest the need to consider in an integrated manner, entomological indicators and environmental management as a condition for effective control actions.

- EPIDEMIOLOGY [1359] P 934 - ASSOCIATION BETWEEN MALNUTRITION AND LEISHMANIA INFECTION: PRELIMINARY RESULTS OF A PROSPECTIVE STUDY IN NORTHERN ETHIOPIA CUSTODIO, E.1; SORDO, L.2; GADISA, E.3; MORENO, J.4; CAÑAVATE, C.4; NIETO, J.4; CHICHARRO, C.4; ASEFFA, A.3; CRUZ, I.4

1.CENTRO NACIONAL DE MEDICINA TROPICAL, INSTITUTO DE SALUD CARLOS III, MADRID, SPAIN; 2.CENTRO NACIONAL DE EPIDEMIOLOGÍA, INSTITUTO DE SALUD CARLOS III, MADRID, SPAIN; 3.AHRI/ALERT ARMAUER HANSEN RESEARCH INSTITUTE, ADDIS ABABA, ETHIOPIA; 4.WHO COLLABORATING CENTER FOR LEISHMANIASIS, INSTITUTO DE SALUD CARLOS III, MAJADAHONDA, SPAIN.

Keyword:visceral leishmaniasis; malnutrition; infection markers

Abstract: Introduction: In 2005 there was a visceral leishmaniasis (VL) out-break in Amhara Regional State, Ethiopia, that has remained with low endemic rates ever since. In this same region protein-energy malnutrition (PEM) is considered a serious public health problem with children malnutrition rates above 50%. The relationship between malnutrition and visceral leishmaniasis is widely accepted, although the underlying mechanisms are still little explained. We conducted a prospective study in the districts affected by the outbreak with the aim to better understand the role of malnutrition in the epidemiology of VL .

Material and Methods: Between May 2009 and February 2011 a cohort study was conducted in Libo Kemkem and Fogera districts (Amhara Regional State). Sampling was carried out by multistaged cluster survey. 522 children negative to Direct Agglutination Test (DAT) and to Leishmanin Skin Test (LST) aged 4 to 15 years were recruited: 52% were classified as malnourished and 48% as non-malnourished. A child was considered malnourished if its Body Mass Index for Age Z score was < -2 Standard Deviations (SD) and/or its Height for Age Z score was < -2 SD according to the WHO 2007 Reference.

Results: During the 21 month follow up, 14 children tested positive for DAT in the malnourished group (cumulative incidence (CI)= 5.1%), and 10 children in the nonmalnourished group (CI =4.0%), being the risk ratio (RR) 1.28 (IC95%=0.58-2.84) . Regarding to LST, at the end of the study, there was one child in the malnourished group (CI=0.37%) and 2 children in the non-malnourished group (CI= 0.8%) that tested positive (RR=0.46; IC 95%=0.04- 5.03).

Conclusions: The preliminary results of this cohort study confirm the low endemicity of VL in the area at the time of the study, and suggest that malnutrition of the host is not associated with an increased vulnerability to infection as measured by DAT or LST. The lower incidence of positivity in LST as compared to DAT is consistent with the longer time required for LST to get positive after infection. Also LST-positive conversion is considered a marker of repeated exposure, which may not be the case in this area of low endemic rates. The association found between not being malnourished and LST positivity could be related to the impaired cellular immune response that occurs in a situation of malnutrition.

- EPIDEMIOLOGY [1366] P 935 - ACCELERATED ECONOMIC DEVELOPMENT IN THE BRAZILIAN AMAZONIAN STATE OF PARÁ INFLUENCES THE EPIDEMIOLOGICAL PROFILE OF CUTANEOUS LEISHMANIASIS CAMPOS, A.L.B.1; ARAÚJO, F.M.1; ARAÚJO, R.S.1; ALBERIO, S.H.V.O.1; GUIMARÃES, S.S.V.2; SOARES, D.C.3; GARCEZ, L.M.4 1.UNIVERSIDADE DO ESTADO DO PARÁ, BELÉM, PA, BRAZIL; 2.SECRETARIA DE ESTADO DE SAÚDE PÚBLICA DO PARÁ, BELÉM, PA, BRAZIL; 3.INSTITUTO EVANDRO CHAGAS AND SECRETARIA DE ESTADO DE SAÚDE PÚBLICA DO PARÁ, BELÉM, PA, BRAZIL; 4.INSTITUTO EVANDRO CHAGAS AND UNIVERSIDADE DO ESTADO DO PARÁ, BELÉM, PA, BRAZIL.

Keyword:cutaneous leishmaniasis; economic development; epidemiological profile

Abstract: The state of Pará (population 7 600 000) has undergone drastic environmental changes in recent years due to accelerated economic development, favoring increased transmission of vectorborne diseases. Pará reports a high incidence of cutaneous leishmaniasis (CL) involving 7 species of Leishmania which cause disease that shows varying levels of resistance to the antimonial Glucantime, the drug of choice for treatment despite its high toxicity. The recent epidemiological profile of CL in Pará is described here. Data from 2007-2011 obtained from the Brazilian system for notification of diseases (SINAN) were used in this cross-sectional descriptive study. Individuals with mucosal lesions, whether associated or not with cutaneous disease, those with leishmaniasis recidivans and those not treated with Glucantime were excluded from the study. Most of the 18 757 cases of tegumentary leishmaniasis were new cases of CL treated with Glucantime (13 297; 71%). Of the 143 municipalities of Pará, 139 reported autochthonous cases of CL. The 5 with the greatest number of cases had populations varying between 27 000-79 000 inhabitants and the respective numbers of cases per 100 000 inhabitants were as follows: Medicilândia 601, Uruará 309, Santarém 44, Tomé-Açú 214 and Tailândia 130.

The predominant epidemiological profile was male (79%), aged 20 and over (70%), of mixed race (70%), low educational level (67%) and rural origin (57%). Indigenous people comprised 1% of the sample. Most (93%) were diagnosed by direct parasitological examination but the remaining 7% did not have access to this test. The antimonial dose (15mg Sb+5/kg/day + 5mg) recommended by the Brazilian Ministry of Health (MS) was used to treat only 86% of the patients. Although the MS recommends Amphotericin B as the preferred medication for pregnant women with CL, 59 were treated with Glucantime. Therapeutic failure was seen in 21%. The description of the recent epidemiological profile of CL, drawing attention to the frequency of 1% among indigenous populations, provides the bases for health interventions geared towards prevention and control of the disease. Such actions should be priorities in the municipalities most affected by the economic development i.e., those close to the metropolitan area of Belém (Tomé-Açu and Tailândia), Tapajós River Basin (Santarém and Uruará) and integration zone of the Xingu river (Medicilândia), where the Belo Monte hydroelectric dam is under construction.

- EPIDEMIOLOGY [1367] P 936 - FACTORS ASSOCIATED WITH CANINE VISCERAL LEISHMANIASIS SEROPOSITIVITY IN THE CITY OF TERESINA (PIAUÍ, BRAZIL): AN APPLICATION USING REMOTE SENSING. GOUVEIA, C.1; ALMEIDA, A.S.1; CRUZ, M.D.S.P.E.2; WERNECK, G.L.1

1.UNIVERSIDADE DO ESTADO DO RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 2.UNIVERSIDADE FEDERAL DO PIAUÍ, TERESINA, PI, BRAZIL.

Keyword:visceral leishmaniasis; canine leishmaniasis; risk factors

Abstract: In the Americas, visceral leishmaniasis (VL) is caused by Leishmania chagasi and transmitted by the sandfly Lutzomyia longipalpis. The domestic dog is considered the main reservoir of the infection, although their role in the transmission cycle is not fully explained. Generally, human cases occur after the canine epidemic. This study aims to evaluate the association of environmental variables with the occurrence of canine infection in the ciy of Teresina (Piauí, Brazil), where the first VL urban epidemic in Brazil was reported. This is a cross-sectional study based on the results of a canine serological survey conducted during 2011 in 10 different localities with moderate or intense transmission of the disease. The survey data were mapped with the aid of GPS. The areas were divided in grids of 900m2 (30m2x30m2) and through remote sensing images, the microenvironment related to types of land cover (water, streets, dark objects, shiny objects, sand or ceramic roof and vegetation) was characterized. Multilevel logistic regression was used to correlate the prevalence of canine infection with environmental variables in the grid where the house was located adjusting for characteristics of the animals (gender, age and race) and the households. 723 dogs were examined distributed in 498 domiciles and 396 grids. A positivity VL rate of about 40% was observed among the participant dogs, with a inter-neighborhood variation from 16 to 54%. The unadjusted analysis, showed an inverse association between canine seropositivity and the area of the grid covered by ceramic roof (p=0.04). Possibly, this protective effect may be related to the improved infrastructure found in these residential areas. However, after adjustment, this effect was not observed. No statistically significant association was found between canine seropositivity and the other environmental variables. In future analyses, other remote sensing variables, such as the NDVI, will be included to improve the ability to explore the data. Studies focusing on the relation between canine VL and the peridomiciliary environment may not only contribute to a greater understanding of the dynamics of the disease in urban areas, but also provide useful information for prevention and control of visceral leishmaniasis in humans.

- CONTROL PROGRAMS [1371] P 937 - DYNAMIC OF VISCERAL LEISHMANIASIS IN POPULATIONS SUBMITTED TO DIFFERENT CONTROL MEASURES SEVÁ, A.P.; GALUTI, E.; OVALLOS, F.G.; GALATI, E.B.; SOARES, R.M.; MORENO, J.; CARRILO, E.; BAQUERO, O.S.; FERREIRA, F. FACULDADE DE MEDICINA VETERINÁRIA - UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL.

Keyword:mathematic model; control; visceral leishmaniasis

Abstract: Visceral Leishmaniasis is a disease that represents serious public health problems to Brazil and some developing countries around the world. The domestic dog is considered the main reservoir of the disease. In Americas the etiologic agent of visceral leishmaniasis is Leishmania infantum chagasi and the transmission of leishmaniasis occurs primarily by the bite of an infected insect sandfly. Current control strategies recommended by the Ministry of Health in Brazil are early

diagnosis and proper treatment of human cases; use of insecticides with residual action and domestic sanitary measures to reduce the vector density; identification and elimination of domestic reservoir. Immunoprophylaxis and use of collars impregnated with deltamethrin in dogs and use of long-lasting insecticidal nets are also important measures to be followed. Due to the difficulty of controlling this disease, cost and complexity of protocols, it is necessary a reevaluation of effectiveness and feasibility of these protocols in theoretical studies, such as the use of mathematical models. Current models do not assess the impact of deltametrinimpreganted collars and the euthanasia for dogs, on visceral leishmaniasis dynamics. In this study we evaluated the evolution of visceral leishmaniasis dynamics with this measures of control alone and in combination, simulating parameters related to hosts and vectors from endemics áreas of Brazil. We could observe that the both, vaccine and collars results in a better effect for a disease control when used together than alone.

- EPIDEMIOLOGY [1372] P 938 - THE INFLUENCE OF VECTOR DENSITY PER HOST ON VISCERAL LEISHMANIASIS DYNAMICS: A SENSITIVITY ANALYSIS BAQUERO, O.S.1; GALVIS, F.O.2; MORENO, J.3; CARRILLO, E.3; GALATI, E.B.2; GALUTI, E.1; SOARES, R.M.1; SEVÁ, A.P.1; FERREIRA, F.1 1.FACULDADE DE MEDICINA VETERINÁRIA E ZOOTECNIA - UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 2.FACULDADE DE SAÚDE PUBLICA - UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 3.INSTITUTO DE SALUD CARLOS III, MAJADAHONDA, SPAIN.

Keyword:vector density; mathematic model; epidemiology

Abstract: The complex interaction among human, dog and vector factors determines the incidence of visceral leishmaniasis (VL). Through mathematical modeling, it is possible to study the influence of each individual factor and assess the effects on the figures of the behavior disease.

However, this approach requires reliable parameters for the populations studied. In some situations, economic and human resources, as well as time constraints, impede the accurate and precise determination of some these parameters. As a result, estimations from other areas are usually used, however the desviation of a single parameter may have profound effects in the analysis and even change the interpretation of the results. The objective of this study was to assess the influence on VL dynamics of variations in both vector density per human and per dog.

For studying the influence of this parameter, we have worked with estimations of vector density, obtained from a literature review. The range of values between the minimum and maximum were modeled using a deterministic approach based on ordinary differential equations. The results showed that variation in the studied parameters have relevant effects on VL epidemiology. The impact of varying vector density per human while fixing vector density per dog was different from the impact resulted from varying vector density per dog while fixing vector density per human. Given this findings, vector density per host must be known precisely, in order to get valid insights of the epidemiology of VL. - EPIDEMIOLOGY [1374] P 939 - SANDFLY FAUNA IN THE OCCURRENCE AREAS OF VISCERAL AND CUTANEOUS LEISHMANIASIS, PIAUÍ, BRAZIL. ARAUJO, I.B.1; GOMES, A.F.S.1; SOUZA, J.V.1; ALBUQUERQUE, E.R.1; MAGALHÃES, A.C.S.1; SILVA, M.L.R.1; MOREIRA, R.M.1; DOURADO, F.V.G.1; GALVÃO, R.F.P.2; SANTOS, O.D.3 1.UNVERSIDADE ESTADUAL DO PIAUÍ, PARNAÍBA, PI, BRAZIL; 2.CENTRO DE ENDEMIAS, PEDRO II, PI, BRAZIL; 3.REGIONAL DE SAÚDE, PARNAÍBA, PI, BRAZIL.

Keyword:sandflies; leishmaniasis; lutzomyia longipalpis

Abstract: Introduction: Leishmaniasis is endemic diseases in Latin America caused by Leishmania and transmitted to man and animals by infected blood-sucking sandflies of the genus Lutzomyia. The Visceral Leishmaniasis (VL) caused by L. chagasi is currently in the process of increasing urbanization and growth in various regions of Brazil with a high prevalence in the state of Piauí one of the main areas of endemic disease. This work aimed to study sandfly fauna and to investigate cases of leishmaniasis in Pedro II and Parnaíba, Piauí. Methodology: Was studied the sandfly fauna as well as cases of leishmaniasis reported in the municipality of Pedro II, period 2005 to 2011, and Parnaíba, period 2007-2011, through the records provided by the Municipal Epidemiological Surveillance of Health. Were captured the sandflies in different city districts, and analyzed sandfly fauna data collected by the Center for Zoonosis Control and Regional Health. Results: Were captured 7.286 sandflies from ten species: Lu. longipalpis, Lu. migonei, Lu. whitmani, Lu. intermedia, Lu.umbratilis, Lu. evandroi, Lu. lenti, Lu. termitophita, Lu.anduzei e Lu. sylvicola, in the period from September/2010 to August/2011 in the city of Pedro II. The most frequent species were Lu. longipalpis with 1972 (27%) specimens collected and Lu. migonei (21.7%). In the period 2005 to 2011 were reported 114 cases of human leishmaniasis in Pedro II, 96 of Cutaneous Leishmaniasis (CL) and 18 of VL, and 119 cases of canine VL. In the municipality of Parnaíba were collected 1.089 sandflies in the períod 20072009, of which 1.040 were Lutzomyia longipalpis species accounting 95.5%. There were 76 reported cases of VL in humans and 453 confirmed cases of the disease in dogs in the investigated period in Parnaíba. In the current study have been verified a large number of cases of canine visceral leishmaniasis and the presence of sandflies in urban areas of Parnaíba.

Conclusion: The present study showed high frequency of sandflies vectors of leishmaniasis with great diversity, abundance and density species in the city Pedro II and dominance of Lu. longipalpis in Parnaíba. It was observed higher prevalence of CL also the occurrence of VL in Pedro II and high frequency de canine visceral leishmaniasis in the municipality of Parnaíba with considerable number of cases of human disease. The data indicate the need for epidemiological surveillance with continuity of control measures particularly of VL in the areas investigated.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1378] P 940 - SPECIES COMPOSITION OF PHLEBOTOMINE SAND FLIES IN A PATCH OF ATLANTIC FOREST IN NORTH-EASTERN BRAZIL SILVA, K.G.1; SILVA, F.J.1; MIRANDA, D.E.O.1; MOURA, A.C.A.1; BRANDÃO-FILHO, S.P.1; DANTAS-TORRES, F.2 1.DEPARTAMENTO DE IMUNOLOGIA, CENTRO DE PESQUISAS AGGEU MAGALHÃES/FIOCRUZ-PE, RECIFE, PE, BRAZIL; 2.DIPARTIMENTO DI MEDICINA VETERINARIA, UNIVERSITÀ DEGLI STUDI DI BARI, BARI, ITALY.

Keyword:sand flies; diversity; ecology

Abstract: Phlebotomine sand flies are vectors of protozoa of the genus Leishmania, the causative agents of leishmaniasis. The ecology of phlebotomine sand flies is of research interest because it can provide background information for a better understanding of the epidemiology of leishmaniasis and to predict the spatial and temporal variations in the risk of infection in a given area. The objective of this study was to identify the diversity of species that make up the phlebotomine sand fly fauna in a remnant of Atlantic Forest located in the Campo de Instrução Marechal Newton Cavalcanti (CIMNC), Paudalho municipality, Pernambuco state, north-eastern Brazil.

From July to December 2012, light traps were placed (ca. 1.5 m above the ground) in 10 sampling sites (P1-P10) near the burrows of wild animals, trunks and roots of large trees in shady and humid environments. Light traps worked from 18:00 until 6:00, for 4 consecutive days each month. The insects captured were initially examined under a stereomicroscope,

separated according to sex and kept in 70% ethanol. The specimens were mounted on slides and identified morphologically. A total of 5226 specimens belonging to 17 species were identified, as follows: Lutzomyia choti (n=3532), L. longispina (n=1095), L. sordelli (n=216), L. complexa (n=184), L. walkeri (n=76), L. amazonensis (n=53), L. naftalekatzi (n=19), L. barrettoi barrettoi (n=16), L. whitmani (n=3), L. wellcomei (n=3), L. capixaba (n=3), L. shannoni (n=1), L. ayrosai (n=3), L. schreiberi (n=2), L. umbratilis (n=14), L. brasiliensis (n=4), and L. viannamartinsi (n=2). The percentage of males (46.5%) captured was lower than that of females (53.5%).

Lutzomyia choti, L. longispina and L. sordelli were the predominant species, being found in all sampling sites. Our findings indicate a high level of phlebotomine sand fly species diversity, revealing the occurrence of L. umbratilis and confirming the presence of L. wellcomei, two proven vectors of Leishmania parasites, in this area.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1379] P 941 - MONTHLY ABUNDANCE OF PHLEBOTOMINE SAND FLIES IN AN AREA OF ATLANTIC FOREST IN NORTHEASTERN BRAZIL MIRANDA, D.E.O.1; SILVA, K.G.1; SILVA, F.J.1; MOURA, A.C.A.1; BRANDÃO-FILHO, S.P.1; DANTAS-TORRES, F.2 1.DEPARTAMENTO DE IMUNOLOGIA, CENTRO DE PESQUISAS AGGEU MAGALHÃES FIOCRUZ-PE, RECIFE, PE, BRAZIL; 2.DIPARTIMENTO DI MEDICINA VETERINARIA, UNIVERSITÀ DEGLI STUDI DI BARI, BARI, ITALY.

Keyword:abundance ; sand flies; vectors

Abstract: The risk of Leishmania transmission is strongly associated with the presence and abundance of phlebotomine sand fly vectors. Information on the seasonal distribution of phlebotomine sand flies is fundamental to predict the risk of Leishmania transmission in a given region.

Nonetheless, information on the monthly dynamics of phlebotomine sand fly vectors is still incipient, particularly in northeastern Brazil. In this context, this abstract reports preliminary data from a study the monthly abundance of phlebotomine sand flies in an area of Atlantic Forest in northeastern Brazil, where cutaneous leishmaniasis by Leishmania braziliensis is endemic. From July to December 2012, phlebotomine sand flies were collected in 10 sites with CDC light traps, which were placed 1.5 m above the ground level near the burrows of wild animals, trunks and roots of large trees, in shaded, humid environments. Each month, traps operated from 18:00 to 6:00 hours during four consecutive nights. The collected insects were kept in 70% ethanol until mounting and morphological identification. Over 4000 phlebotomine sand fly specimens belonging to 17 species were collected. The overall monthly average of phlebotomine sand flies was 871 (range, 593-1173; 87.1 sand flies per trap), with peaks in September (106.8 sand flies per trap), October (117.3 sand flies per trap) and December (107.3 sand flies per trap). Interestingly, L. choti peaked in September, October and December, when a total of 2372 specimens were collected. So far, our results suggest a high abundance of phlebotomine sand flies in the study area, with a peak in collections at the end or after the raining season. We will continue to follow up this phlebotomine sand fly population monthly for two years and data generated will provide valuable information regarding the seasonal dynamics of several species, including proven Leishmania spp. vectors. Indeed, this will help us to predict the period in which there may be an increased risk of Leishmania transmission in remnants of Atlantic Forest.

- RESERVOIRS [1380] P 942 - ROLE OF M. SHAWI IN THE SPREAD OF ZCL IN CENTRAL TUNISIA USING TELEMETRY AND MATHEMATICAL MODELING. ZAATOUR, W.; CHLIF, S.; GHAWAR, W.; BETTAIEB, J.; SNOUSSI, M.A.; BELHAJ HMIDA, N.; SALAH, A.B.

INSTITUT PASTEUR DE TUNIS, TUNIS, TUNISIA.

Keyword:m shawi, telemetry; fickian diffusion; spatio-temporal dynamics

Abstract: A study was undertaken between March and December 2012, to evaluate the contribution of Meriones (M.) shawi to the geographic spread of the zoonotic cutaneous leishmaniasis (ZCL).

We had quantified the spatial dynamic of the rodent by telemetry. 30 M. shawi were equipped

by VHF radio collars and were tracked for 6 months. We determined their average Home Range (HR) (0.1507 Km²) and the average traveled distance (0.734 Km). We estimated the per capita rate of ZCL infection among M. shawi sample from previous study (3.1 Indivdual-1 Year-1) and the diffusion coefficient (0.6024 Km²Year-1). A spatial mathematic model based on Fickian Diffusion was developed to describe the M. shawi spatial-temporal dynamics. Different simulation scenarios demonstrated its high mobility which accelerates ZCL infection among rodents. To our knowledge it is the first time the dynamics of M. shawi is quantified in Tunisia.

It contributes to further incriminate M. shawi in the spread of ZCL among Humans.

- EPIDEMIOLOGY [1382] P 943 - TEGUMENTARY LEISHMANIASIS ASSOCIATED WITH HIV INFECTION IN THE BRAZILIAN AMAZONIAN STATE OF PARÁ MIRANDA, H.C.1; TAVARES, L.H.M.1; ALBERIO, S.H.V.O.1; SOARES, D.C.2; GARCEZ, L.M.3 1.UNIVERSIDADE DO ESTADO DO PARÁ, BELÉM, PA, BRAZIL; 2.INSTITUTO EVANDRO CHAGAS E SECRETARIA DE ESTADO DE SAÚDE PÚBLICA DO PARÁ, BELÉM, PA, BRAZIL; 3.INSTITUTO EVANDRO CHAGAS E UNIVERSIDADE DO ESTADO DO PARÁ, BELÉM, PA, BRAZIL.

Keyword:tegumentary leishmaniasis; hiv; amazonian

Abstract: In the state of Pará (population 7 600 000), the epidemiology of the tegumentary leishmaniasis (TL) associated with HIV infection needs to be known, given the increasing economic development of the state. Pará shelters 13 regional health centres (RHC) strategically situated for covering the state’s full geographic extension. The recent epidemiological profile of TL/HIV in Pará is described here. Data from 2007-2011 obtained from the Brazilian system for notification of diseases (SINAN) were used in this cross-sectional descriptive study. Individuals with both TL (any clinical form) and HIV infection (whether having AIDS or not), being autochthonous cases from Pará, males and females of all ages were included. Of the 18 771 TL cases reported in the period, 0.5% (98) had HIV infection. Most were new cases of co-infection (90%), recurrent lesions being seen in the other 10%. The predominant epidemiological profile was male (80%), aged 20-50 (63%), of mixed race (62%) and low educational level (66%).

Indigenous people and children aged 10 years or less comprised 3% and 8% of the sample, respectively. Most (91%) had cutaneous lesions, being reported mucosal (4%) and mucocutaneous (5%) disease for the others. Fields indicating other clinical presentations (diffuse or disseminated lesions) were not available in SINAN. Cure was attained in most cases (62%), drawing attention to the lack of this information in 30% of the records. Most of the TL/HIV cases were reported by RHCs of the most dynamic regions in the state: 35 in Carajás region (RHC 11, Marabá) and 23 in Tapajós River Basin (RHC 9, Santarém). Other municipalities from both regions reporting high number of TL/HIV cases in recent years were Almeirim, Novo Progresso and Eldorado dos Carajás (8, 6 and 10 cases, respectively). An annual average of 3 800 TL cases was registered during 2007-2011. In addition, HIV heterosexual transmission is increasing in Pará, where is currently intense the migration phenomenon. Therefore, in despite of the higher frequency of TL/HIV infection in men adults, the incidence in women and children must be investigated and followed up, specially in the southern (Carajás) and western (Tapajós) of Pará. The surveillance for both diseases should

enhance in indigenous communities. The form used for TL notification must be reviewed in order to make the filling easier and to include fields for registering other clinical presentations of the disease.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1390] P 944 - CONTRIBUTION TO THE KNOWLEDGE OF THE EPIDEMIOLOGY OF LEISHMANIOSIS IN THE NE OF SPAIN THROUGH THE STUDY OF THE ALTITUDINAL AND BIOCLIMATIC DISTRIBUTION OF THE VECTORS, PHLEBOTOMUS ARIASI AND P. PERNICIOSUS, IN LLEIDA (CATALONIA) BALLART, C.1; GUERRERO, I.2; CASTELLS, X.3; BARÓN, S.4; ALCOVER, M.M.1; CASTILLEJO, S.1; PORTÚS, M.5; GÁLLEGO, M.6 1.LABORATORI DE PARASITOLOGIA, FACULTAT DE FARMÀCIA, UNIVERSITAT DE BARCELONA; CRESIB, BARCELONA, SPAIN; 2.DEPARTAMENTO DE ECOLOGÍA, FACULTAD DE CIENCIAS, UNIVERSIDAD AUTÓNOMA DE MADRID, MADRID, SPAIN; 3.DSERVEI DE GENÒMICA I BIOINFORMÀTICA, PARC DE RECERCA UAB MRB-IBB, CERDANYOLA DEL VALLES, SPAIN; 4.EDEPARTAMENTO DE PARASITOLOGÍA, FACULTAD DE FARMACIA, UNIVERSIDAD DE GRANADA, GRANADA, SPAIN; 5.LABORATORI DE PARASITOLOGIA, FACULTAT DE FARMÀCIA, UNIVERSITAT DE BARCELONA, BARCELONA, SPAIN; 6.LABORATORI DE PARASITOLOGIA, FACULTAT DE FARMÀCIA, UNIVERSITAT DE BARCELONA, AV JOAN XXIII ; CRESIB, BARCELONA, SPAIN.

Keyword:phlebotomus ariasi; phlebotomus perniciosus; ne spain

Abstract: In Spain, leishmaniosis is caused by Leishmania infantum and two sand flies species, Phlebotomus ariasi and P. perniciosus, are the proven vectors living in sympatric conditions in several foci. In recent years, an increase in canine leishmaniosis has been observed in central and southern Spain as well as the presence of the disease in northern areas including the Pyrenean region of Lleida province The aim of the present study was to determine the role of altitude and bioclimatic zone in the distribution and density of each of the L. infantum vectors in this province.

Lleida is situated in Catalonia (NE of Spain), bordering to the north with the Pyrenean areas of Andorra and France. It covers an extension of 12,173km2. An entomological study was carried out in July 2006, with sticky traps set in sand flies diurnal resting places and recovered after four days, in the entire province at altitudes ranging from 86 to 1755 meters a.s.l. (339 sites were sampled). The results were analysed using bivariate analysis.

A total of 12,777 sand flies were captured and five species were identified with the following densities (specimens/m2 sticky trap): 32.5 of Sergentomyia minuta, 4.5 of P. perniciosus, 1 of P. ariasi, 0.8 of P. papatasi and 0.2 of P. sergenti. The bivariate analysis revealed that altitude and bioclimatic zone were significantly associated with sand fly densities in opposite way, according to the vector species. The density of P. perniciosus was negatively correlated with altitude (continuous variable IRR 0.99 and >800m IRR: 0.21), whilst the density of P. ariasi was positively correlated (continuous IRR: 1.003 and >800m IRR: 4.98). According to these results, the distribution of the vectors showed also an opposite bioclimatic distribution: P. perniciosus was positively correlated with: bioclimatic zones (supra-Mediterranean IRR: 2.90 and mesoMediterranean IRR: 3.93) whilst P. ariasi was negatively correlated (supra-Mediterranean IRR: 0.43 and meso-Mediterranean IRR: 0.02). The results emphasises the importance of the individual analysis of vectors living in sympatric conditions in studies on the risk of leishmaniosis transmission.

This work was supported by grants of the Ministerio de Educación y Ciencia of Spain (AGL2004-06909-C02-01), Departament d’Universitats, Recerca i Societat de la Informació de la Generalitat de Catalunya (Spain) (2009SGR385) and European Union (GOCE-2003-010284 EDEN, Emerging Diseases in a changing European eNvironment).

- EPIDEMIOLOGY [1393] P 945 - ACTIVITIES OF VISCERAL LEISHMANIASIS CONTROL IN DOGS IN AN AREA ENDEMIC OF BELO HORIZONTE CITY, MINAS GERAIS STATE, BRAZIL, FROM 2006 TO 2011.

GONÇALVES, S.A.1; GUIMARÃES, E.L.A.M.1; OLIVEIRA, M.A.1; FREITAS, A.C.P.1; SANTOS, M.R.A.2; MORAIS, M.H.F.2; ARAÚJO, V.E.M.2; SILVA, J.A.1; SOARES, D.F.M.1 1.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE, MG, BRAZIL; 2.SECRETARIA MUNICIPAL DE SAÚDE, BELO HORIZONTE, MG, BRAZIL.

Keyword:leishmaniasis; dog; brazil

Abstract: Belo Horizonte is an endemic municipality for visceral leishmaniasis that presents taxes of incidence and letality superiors of those found in Minas Gerais state and in Brazil. It was made a descriptive study about the activities of control of the disease in dogs, in one region of the city, from 2006 to 2011, with data obtained from the Information System for Zoonosis Control provided by the Municipal Secretariat of Health. From a total of 151,725 tests, 86.1% were negative, 7.4% were positive, 2,1% were undetermined (inconclusive in the indirect immunofluorescence test) and 3.8% were monitor (positive in the immunoenzymatic assay and negative in the immunofluorescence test). There was an increase of the coverage of census surveys and a decrease in the tests requested by residents, followed by decreasing of the canine seropositivity in areas covered in the period. Undeterminated results also have declined, while serologies "monitor" has increased. The seroconversion rate among dogs undetermined was 54.3%, occurring in 104 days on average, with median of 54 days. It was found that 98, 1% of the samples were collected on filter paper and that the laboratory diagnosis was delivered in 18.1 days on average, after blood collection. The infected dogs removal happened in 51.7 days on average, with median of 39 days, counted since the obtention of the sample. The percentage of withdrawal of soropositive dogs was 86.6%, which was negatively impacted by refusals, which represented 7.8% over the period, ranging from 1.9% to 15.6%. The database from the Information System for Zoonosis Control of Belo Horizonte was conducive to study the expansion and the combat of the disease in the city. Thus, it is recommended that it receives periodic maintenance of its quality, so that further studies might be performed. It is also necessary to intensify the activities of health education in the municipality in order to alert citizens about the dog’s importance in the infection transmission dynamics and thereby increase the population adherence to the visceral leishmaniasis control program.

Apoio: Pró-Reitoria de Pesquisa da Universidade Federal de Minas Gerais, CAPES, FAPEMIG, CNPq, Secretaria Municipal de Saúde de Belo Horizonte.

- EPIDEMIOLOGY [1394] P 946 - STUDIES ON THE PREVALENCE AND CHARACTERIZATION OF LEISHMANIA SPECIES IN GHANA BATES, P.A.1; KWAKYE-NUAKO, G.2; MOSORE, M.3; BATES, M.D.1; DESEWU, K.4; PUPLAMPU, N.5; ATTIPOE, I.6; AYEH-KUMI, P.6; JAMJOOM, M.1; BOAMPONG, J.2; DUPLESSIS, C.5; BOAKYE, D.3 1.LANCASTER UNIVERSITY, LANCASTER, UNITED KINGDOM; 2.UNIVERSITY OF CAPE COAST, CAPE COAST, GHANA; 3.NOGUCHI MEMORIAL INSTITUTE FOR MEDICAL RESEARCH, ACCRA, GHANA; 4.GHANA HEALTH SERVICE, ACCRA, GHANA; 5.US NAVAL MEDICAL RESAERCH UNIT 3, GHANA DETACHMENT, ACCRA, GHANA; 6.UNIVERSITY OF GHANA MEDICAL SCHOOL, ACCRA, GHANA.

Keyword:ghana; cutaneous leishmaniasis; identification

Abstract: Leishmaniasis is an emerging or re-emerging disease in many parts of the world, especially in developing countries. In Ghana, until recently leishmaniasis was relatively unknown and little work has been done on the disease. However, since 1999 reports of cutaneous leishmaniasis in the Volta region of the country have been made, with suggestions of a high prevalence in the affected area. The identity of the aetiological agent(s) remains to be firmly established, although L. major has been previously suggested together with an unidentified species. Further work is required to confirm these findings and explore the relationship of the Ghanaian parasites with Leishmania from elsewhere in Africa. In this study we aimed to identify the species of Leishmania responsible for leishmaniasis infections in the Ho district of the Volta region of Ghana. Patients presenting with cutaneous lesions suggestive of cutaneous leishmaniasis in several rural communities in the Ho district were recruited into the study. A total of 44 samples were initially examined, which comprised either skin scrapings or aspirates collected from the sites of active lesion(s), using a single-use sterile scalpel and needle with syringe, respectively.

These were collected on to FTA cards in the field and transported to the laboratory for molecular characterisation using PCR assays. The presence of Leishmania was confirmed using genus-specific minicircle (kDNA) and ribosomal primers in 36 and 27 of the samples, respectively. Recently parasites were successfully isolated into culture for the first time in Ghana, and these are currently undergoing further analysis by DNA sequencing to reach a firm conclusion regarding their identity, a result we expect to report at Worldleish5. Ongoing studies to identify the vector responsible for disease transmission and potential reservoir hosts are also underway.

- EPIDEMIOLOGY [1395] P 947 - THE EPIDEMIOLOGY OF CANINE LEISHMANIOSIS IN THE BALEARIC ISLANDS (SPAIN): VETERINARY QUESTIONNAIRE AND PROSPECTIVE CANINE LEISHMANIOSIS SURVEY ALCOVER, M.M.1; BALLART, C.1; SCALONE, A.2; SERRA, T.3; RIERA, C.4; GRAMICCIA, M.2; PORTÚS, M.4; GÁLLEGO, M.5 1.LABORATORI DE PARASITOLOGIA, FACULTAT DE FARMÀCIA, UNIVERSITAT DE BARCELONA; CRESIB, BARCELONA, SPAIN; 2.SECTION OF VECTOR-BORNE DISEASES AND INTERNATIONAL HEALTH, MIPI DEP, ISTITUTO SUPERIORE DI SANITÀ, ROMA, ITALY; 3.INSTITUT UNIVERSITARI D'INVESTIGACIÓ EN CIÈNCIES DE LA SALUT (IUNICS), CAMPUS UNIVERSITARI, PALMA DE MALLORCA, SPAIN; 4.LABORATORI DE PARASITOLOGIA, FACULTAT DE FARMÀCIA, UNIVERSITAT DE BARCELONA, BARCELONA, SPAIN; 5.LABORATORI DE PARASITOLOGIA, FACULTAT DE FARMÀCIA, UNIVERSITAT DE BARCELONA, AV. JOAN XXIII; CRESIB, BARCELONA, SPAIN.

Keyword:canine leishmaniasis; balearic islands; minorca

Abstract: Leishmaniosis is present in the Mediterranean region of Europe where Leishmania infantum is responsible of the disease. Dogs are the domestic reservoir but, despite the recommendations of the OIE, there is not official data about the distribution and prevalence of the disease (CanL) in Spain. The data published from the Balearic Islands showed a rate of infection of 0.9%-77% according to the island, the kind of dog population analyzed and the diagnostic technique used.

Studies carried out during the eighties showed that, unlike the other islands, the prevalence of CanL in Minorca was very low (0.9%), and all the cases detected were considered imported.

Despite this situation, entomological studies demonstrated the presence of P. perniciosus in the island of Minorca. In the last time several studies reported the emergence or reemergence of leishmaniosis in the Mediterranean foci. This induced us to carry out an epidemiological study on CanL in the Balearic Islands to establish its actual situation.

A veterinary questionnaire was sent out to 111 veterinary clinics. 42 of the questionnaires were returned completed (reply rate of 38%). Over 80% of surveyed veterinary clinics attended more than 20 dogs a week, from which 73.8% had more than 10 dogs confirmed of having CanL

during the last 12 months. Whilst a 50% of the veterinarians thought that CanL has not changed over time other 26.2% perceived an increasing trend, mainly those from Minorca. Most veterinarians considered the new cases diagnosed being autochthonous (88.1%). Due to the results obtained, a cross sectional study on CanL was performed in Minorca by serological methods on 121 dogs of three veterinary clinics located so as to cover different areas of the island. 29 out of them (24%) were seropositive, ranging the seropositivity from 12 to 40% depending on the veterinarian clinic. The seroprevalence between animals we know the local origin and having no history of movements to endemic areas (69 dogs) was 31%. The previous knowledge of the disease in the island of Minorca and the results obtained through the veterinary questionnaire as well as in the prospective study let us to conclude that the presence of CanL in Minorca is consequence of an emergence.

Work supported by Grants CGL2007-66943-C02-01/BOS (Ministerio de Ciencia y Tecnología, Spain) and Departement d’Universitats, Recerca i Societat de la Informació de la Generalitat de Catalunya (Spain) (2009SGR385). EDEN-LEI allowed the work of MMA in Rome

- EPIDEMIOLOGY [1396] P 948 - SOCIO-ENVIRONMENTAL CHARACTERISTICS AND THE OCCURRENCE OF AMERICAN CUTANEOUS LEISHMANIASIS VECTORS OF IN THE CITY OF RIO DE JANEIRO, BRAZIL SENA, C.A.P.1; DE AZEVEDO, A.C.R.2; ALMEIDA, A.S.3; WERNECK, G.L.4 1.ESCOLA NACIONAL DE SAUDE PUBLICA, RIO DE JANEIRO, RJ, BRAZIL; 2.INSTITUTO OSVALDO CRUZ, RIO DE JANEIRO, RJ, BRAZIL; 3.DEPARTAMENTO DE ENDEMIAS SAMUEL PESSOA ESCOLA NACIONAL DE SAÚDE PÚBLICA, RJ, RJ, BRAZIL; 4.FUNDAÇÃO OSWALDO CRUZ INSTITUTO DE MEDICINA SOCIAL – UERJ)., RIO DE JANEIRO, RJ, BRAZIL.

Keyword:lta; sensoriamento remoto; rio de janeiro

Abstract: Socio-environmental characteristics and the occurrence of American cutaneous leishmaniasis vectors of in the city of Rio de Janeiro, Brazil Caroline Almeida Pereira Sena 1, Alfredo Carlos Rodrigues de Azevedo 2, Andréa Sobral de Almeida 3, Guilherme Loureiro Werneck 4 1 Mestranda Epidemiologia em Saúde Publica - Escola Nacional de Saúde Publica ENSP/FIOCRUZ; 2 Laboratório de Transmissores de Leishmanioses/IOC; 3 Departamento de Endemias Samuel Pessoa Escola Nacional de Saúde Pública 4Fundação Oswaldo Cruz Instituto de Medicina Social – UERJ).

The resurgence of diseases such as leishmaniasis reflects an increasing anthropic modification of the environment and is manifested by great regional heterogeneity. In this context, the use of environmental approaches through the use of remote sensing enables new perspectives for the study of leishmaniasis, allowing a better comprehension of the socio-environmental vulnerabilities that contribute to disease transmission. The aim of this study was to evaluate the relationship between socio-environmental characteristics and the occurrence of vectors of American cutaneous leishmaniasis (ACL) in neighborhoods of the city of Rio de Janeiro, Brazil.

Eight localities were selected for weekly captures of sandflies. In each neighborhood light traps were placed three times in the peridomicile of six different houses (144 collections in 48 houses). These traps were georeferenced using Global Positioning System. We carried out analysis of land cover using information extracted from a Landsat 5 TM remote sensing image.

Between the months of March to September 2012, 479 specimens of phlebotomines were

collected belonging to ten species and two genera, Brumptomyia and Lutzomyia. L. intermedia (88,8%) was the predominant specimen followed by L. migonei (6%). The presence of vectors of leishmaniasis were found in 15.3% of the traps. The areas with the greatest abundance of vectors of leishmaniasis were those located in regions with Normalized Difference Vegetation Index (NDVI) between average (20%) and low (19%) values. While L. migonei concentrated in points with low, medium and high NDVI, L. intermedia concentrated only in areas with low and mean NDVI. When captures were stratified by phases of the moon, it was observed that in the new moon there was a greater proportion of points with vectors captured (19.44%), as compared to periods of full (12.50%) and waning (14.93%) moons. The local where traps were placed with greater occurrences of vectors were kennel (33%), chicken sheds (14%), banana / guava / coconut trees (12%) and porch / garage (11%). We observed a statistically significant (p = 0.01) association between the occurrence of L. migonei and the location of traps. The results of this study show that some characteristics of the local environment may be associated with an increased risk for ACL and should be considered when prioritizing areas for intervention.

- CONTROL PROGRAMS [1397] P 949 - LEISHMANIA IDENTIFICATION ON CLINICAL SAMPLES FROM CUTANEOUS LEISHMANIASIS PATIENTS IN THE AMAZONIAN STATE OF PARÁ AND THE CHALLENGES TO REDUCE MORBIDITY CAUSED BY THE ANTIMONIAL TREATMENT SOARES, D.C.1; MARTINS, N.V.D.N.2; SILVA, F.L.D.2; MOTA, M.D.S.S.2; QUIROGA, M.M.2; CHAGAS, A.P.3; SAGICA, F.D.E.S.3; SHAW, J.J.4; CUPOLILLO, E.5; DOS SANTOS, L.M.G.6 1.INSTITUTO EVANDRO CHAGAS/SECRETARIA DE ESTADO DE SAUDE PÚBLICA DO PARÁ, ANANINDEUA, PA, BRAZIL; 2.UNIVERSIDADE DO ESTADO DO PARÁ, SANTARÉM, PA, BRAZIL; 3.INSTITUTO EVANDRO CHAGAS, ANANINDEUA, PA, BRAZIL; 4.UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL; 5.INSTITUTO OSWALDO CRUZ/FIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 6.INSTITUTO EVANDRO CHAGAS/UNIVERSIDADE DE ESTADO DO PARÁ, BELEM, PA, BRAZIL.

Keyword:etiology of leishmania ; cutaneous leishmaniasis; state of pará

Abstract: The state of Pará registered an annual average of 3 800 tegumentary leishmaniasis (TL) cases during 2007-2011. In this Amazonian area the seven Leishmania species known to cause human disease could show varying levels of resistance to the antimonial Glucantime. The subgenus Viannia, especially Leishmania (Viannia) braziliensis is frequently associated to antimonial resistance. We describe the etiology and the response to antimonial treatment in a cutaneous leishmaniasis (CL) case series. Patients (77) were men (69) and women (8), aged 10-73, having CL for the first time. The study was carried out in Juruti (n = 18; 2007-2009) and Santarém (n = 59; 2009-2011) municipalities. The individuals were followed during Glucantime therapy (15mg SB+5/kg/day/20 days) and until three months after treatment for investigating complete reepithelization (cure). PCR was performed for detecting Leishmania species in skin biopsies using glucose-6-phosphate dehydrogenase (G6PD), restriction fragment length polymorphism of ribosomal internal transcribed spacers (RFLP ITS) and of heat-shock protein 70 (RFLP Hsp-70).

The patients were infected with L. (V.) braziliensis (53%), Leishmania (V.) sp (40%) and L. (Leishmania) amazonensis (7%). Only 40 individuals could be followed up as 37 abandoned the treatment. The two-tailed binomial test at the 0.05 level of significance was performed. Most of the patients cured after treatment (67,5%). The proportion of cured patients starting the treatment one or two months after arising the primary lesion was similar, respectively 60% and 72% (Z = 0,7845; p = 2164) and was also the same between patients infected with L. (V.) braziliensis (n = 22; 73%) or L. (V.) sp (n =18; 61%), since there was no statistical difference (Z = 0.7803; p = 0,2176). Patients with more than one primary lesion presented lower rate of cure (n = 10; 40%) if compared to those having unique lesion (n= 30; 76%) (Z = 2.1439; p= 0,0160).

Among the patients who abandoned treatment, 48% were infected with L. (V.) braziliensis. The high proportion of CL caused by L. (V.) braziliensis, the difficulties to perform the regular clinical follow-up for CL patients and the high frequency of multiples lesions in primary infection can lead to an increase in morbidity caused by the repetition of the therapy.

- RESERVOIRS [1398] P 950 - LEISHMANIOSIS IN DOGS IN CAJUﾂｴS NEIGHBORHOOD QUINTﾃグ, L.C.; GOMES, K.; BORGES, D.T.; DUTRA, J. SMSDC - DVS - CAP 1.0, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:dog ; caju; phlebotomus

Abstract: Was notified on 05.26.11 a case of canine visceral leishmaniasis, a mixed breed, female, who lived until November 2010 at the kennel Cemetery St. Francis Xavier with 29 other dogs and from this date he moved to a residence in Caju same neighborhood, an area hitherto considered non-endemic for Leishmania, the site had about 23 more dogs. The notifications were made by Jorge Vaitsmam Institute, which reported that the dog in question was brought to the IJV demand for the clinic's owner, and then having the material sent for examination was positive.

Because then the presence of these animals showing the disease was made a search for detecting the presence of Phlebotomus sp. In the region and both vectors were found in the cemetery and in the region of the residences. So following the technical standards was made a research in all areas of the neighborhood and confirmed the presence of the vector Lutzomyia longipalpis infesting most areas surveyed.

Both kennel in the cemetery as the residence dogs were examined and diagnosed as positive for leishmaniasis, these animals were all referred to the sacrifice. With the high positivity began to make an active search dogs in areas near the locations of the two kennels and once again found positive cases, including dogs that lived inside and residence within a military installation located adjacent to the said cemetery.

So the service of the environmental monitoring DVS-CAP 1.0 has treated as an endemic area for leishmaniasis and has been monitoring the area since then. This monitoring is done to investigate the presence of vectors and animals that may be involved as reservoirs of the disease.

With the region being considered endemic to the family health team that is responsible for the neighborhood Caju, Family Clinic Fernando Lopes Braga, was trained and sensitized by service excellence Research Institute Evandro Chagas to recognize and diagnose potential cases leishmaniasis in humans.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1399] P 951 - GENUS LUTZOMYIA (DIPTERA, PSYCHODIDAE, PHLEBOTOMINAE) IN AREAS WITH TRANSMISSION OF LEISHMANIASIS: NATURAL BREEDING SITES AND UTILITY OF DNA TAXONOMY AND BARCODING FOR THE IDENTIFYING OF IMMATURE MUSKUS, C.E.1; VIVERO, R.J.1; CONTRERAS, M.A.1; BEJARANO, E.E.2; GUTIEREZ, C.T.1; URIBE, S.I.3 1.PROGRAM FOR THE STUDY AND CONTROL OF TROPICAL DISEASES, MEDELLIN, COLOMBIA; 2.BIOMEDICAL GROUP, SUCRE, COLOMBIA; 3.MOLECULAR SYSTEMATICS GROUP, MEDELLIN, COLOMBIA.

Keyword: lutzomyia; breeding sites; control

Abstract: Abstract text: The taxonomic determination of genus Lutzomyia is fundamental in biological studies of the leishmaniasis control in America. By taking into account the poor knowledge

breeding sites of phlebotomine in leishmaniasis transmission areas, and the absent of taxonomic keys for immature stages. In nowadays, the entomological analysis of genus Lutzomyia is mainly based on adult insects, without taking into account the immature stages, suggesting the existence of incomplete inventories. For these reasons, it is fundamental to develop new schemes and complementary analysis that support the taxonomic classification.

Objective: Detect natural breeding sites of genus Lutzomyia and assign taxonomic identity of immatures isolated with classical and molecular tools.

Methodology: The samples were collected in leishmaniasis transmission areas in the Colombian provinces of Antioquia, Chocó, and Sucre. We worked in potential 160 breeding sites. The larvae/pupae differentiation was based on direct observation through stereoscope. Each of the individuals that were found alive in the samples was reared under laboratory conditions until the adult stage. Subsequently identified by using the taxonomic key of Young and Duncan, 1994.

The larvae or pupae that died during the process were genetically identified with the mitochondrial marker COI (Kimura-2P genetics distance and NJ tree).

Results: 142 immature were isolated in 38 breeding sites. Immatures were found mainly in roots (n= 51; 35.91%) and bases of trees (n= 35; 24.64%;). We identified 13 species of Lutzomyia and two of Brumptomyia. A number of 60 inmature (42.25%) were identified by classical taxonomy (adults emerged). Taxonomic identity of 18 inmature (12.67%) was assigned with COI gene analysis, which revealed a intraspecific divergence K2P genetic of 0.002 to 0.033 and values bootstrap (> 95).

Conclusions: Cytochrome Oxidase I (barcoding DNA), facilitates the identification of immature of genus Lutzomyia indistinguishable morphologically (species level). It is a cost effective tool, of use simultaneous for reveal the identity of the insects vectors transmitters of leishmaniasis.

- CONTROL PROGRAMS [1400] P 952 - CURRENT EPIDEMIOLOGICAL PROFILE OF VISCERAL LEISHMANIASIS AND FACTORS ASSOCIATED WITH LETHALITY IN PARÁ STATE, BRAZIL SANTOS, M.B.L.1; TUMA, K.S.1; COSTA, J.L.F.1; FADUL, M.R.2; SOARES, D.C.3; CHAGAS, A.P.4; GARCEZ, L.M.5 1.UNIVERSIDADE DO ESTADO DO PARÁ, BELÉM, PA, BRAZIL; 2.SECRETARIA DE ESTADO DE SAÚDE PÚBLICA DO PARÁ, BELÉM, PA, BRAZIL; 3.INSTITUTO EVANDRO CHAGAS AND SECRETARIA DE ESTADO DE SAÚDE PÚBLICA DO PARÁ, BELÉM, PA, BRAZIL; 4.INSTITUTO EVANDRO CHAGAS, BELÉM, PA, BRAZIL; 5.INSTITUTO EVANDRO CHAGAS AND UNIVERSIDADE DO ESTADO DO PARÁ, BELÉM, PA, BRAZIL.

Keyword:visceral leishmaniasis; lethality; epidemiological profile

Abstract: The state of Pará reported high rates of lethality (3%-6%) caused by visceral leishmaniasis (VL) from 2001-2010. The aspects associated to the deaths vary in different endemic areas and need to be known for improving locally the surveillance and prevention. We describe clinical and epidemiological aspects, highlighting those associated to lethality in the state of Pará. Data from 2007-2011 obtained from the Brazilian system for notification of diseases (SINAN) were used in this cross-sectional descriptive study. Individuals with VL, being autochthonous cases from Pará, males and females of all ages were included. Of the 1 771 VL cases reported in the period, 3.5% (62) died, 56% (993) cured, 13.5% (240) had no information and 27% (476) died for other reasons or had been referred to another hospital. In despite of the gradual reduction in the number of VL cases yearly reported, the disease detection rates (4.1-5.6) and lethality (2.5-4.2) did not vary significantly (p>0.05) in 5 years. Of the 143 municipalities of Pará, 91 (63%)

reported autochthonous cases of VL, being areas of high (20), low (6) and moderate (65) transmission. The municipalities where VL is highly endemic had the highest disease detection rates (9.5-13%) and the lowest lethality (2-4.5%) if compared to those of low (1.7-6.6%; 0-12%) and moderate transmission (0.8-1; 3-18%), respectively. The predominant epidemiological profile was male (59%), aged 4 years or less (49%), of mixed race (77%) and rural origin (65%).

Among those who died, 39% aged 1 year or less and 65% had been treated with Glucantime.

Signs indicating severity and poor prognosis were also seen in this group in the following rates: edema (58%); other infectious process (37%) and hemorrhages (29%). The evolution time to death after the therapeutic intervention was two months or less in 77% patients. Given the lack of information related to the outcome after treatment in SINAN database, the lethality in Pará must be higher than 3,5%. In the municipalities where VL transmission is low or moderate the early diagnosis of the disease must be less frequent, since the lethality is higher in these areas if compared to those with high transmission rates. The description of the recent epidemiological profile of VL provides the bases for health interventions geared towards prevention and control of the disease, but it is still necessary to investigate other social and biological factors, including co-morbidities, influencing lethality.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1401] P 953 - ENTOMOLOGICAL STUDY TO SURVEY THE RISK OF HUMAN INFECTION FOR CUTANEOUS LEISHMANIASIS IN THE BOUNDARY ZONE BETWEEN THE RURAL AND URBAN ÁREAS OF ILHEUS, BA CARVALHO, S.M.S.1; SANTOS, P.R.B.2 1.UNIVERSIDADE ESTADUAL DE SANTA CRUZ, ILHÉUS, BA, BRAZIL; 2.DIRETORIA REGIONAL DE SAÚDE, ILHÉUS, BA, BRAZIL.

Keyword:leishmaniasis; lutzomyia; ilhéus

Abstract: This research proposed to accomplish entomological study in the boundary zone between the rural and urban areas of the city of Ilheus, BA, known as Mata da Esperança, to evaluate human exposure in contact with insects of the genus Lutzomyia. Captures were carried with a CDC light trap and Castro trap, and were collected 214 sand flies between March/2010 and December/2011. Specimens were identified morphologically aiming to identify sandflies in the city. Five species were described: Lutzomyia fischeri (52.8%), L. choti (37.9%), L. whitmani (5.6%), L. viannamartinsi (2.3%) and L. davisi (1.4%). Low density of insects found shows that it is unlikely to maintain a zoonotic cycle around the urban area studied, different than suggested by reports of human cases. It ´s believed in a rural origin of the cases identified in the urban área. Moreover, the proximity of the Mata da Esperança with the rural environment justifies the finding of insects in this border area, but apparently there isn’t risk to the urban area.

- OPERATIONAL HEALTH [1403] P 954 - STRATIFICATION OF HOUSEHOLDS ACCORDING TO THE RISK OF PHLEBOTOMINE SANDFLIES INFESTATION BY LOCAL HEALTH AGENTS IN THE CITY OF BELO HORIZONTE, MINAS GERAIS, BRASIL VIANNA, E.N.1; MORAIS, M.H.F.2; ALMEIDA, A.S.3; SABROZA, P.C.3; DIAS, E.S.4; CARNEIRO, M.1

1.UFMG, BELO HORIZONTE, MG, BRAZIL; 2.MUNICIPAL HEALTH DEPARTMENT, BELO HORIZONTE, MG, BRAZIL; 3.ENSPFIOCRUZ, RIO DE JANEIRO, RJ, BRAZIL; 4.CPQRR FIOCRUZ-MINAS, BELO HORIZONTE, MG, BRAZIL.

Keyword:stratification; phlebotomine sandflies; house infestation

Abstract: Among Latin American countries, Brazil has the largest number of cases of human visceral leishmaniasis (LVH). In Belo Horizonte, the disease occurs since 1994, where is largely dispersed. The vector Lutzomyia longipalpis may be present in peridomicile and intradomicile.

Thus, enabling the identification of the receptive properties (a lot of organic matter, fruit trees, moist and shady places, host availability) and vulnerable ones (presence of attic, crevices, cracks, broken windows) by the endemic disease combat agents (ACE), can help the Visceral Leishmaniasis Control Program to direct their activities. The aim of this study was to test the methodology for identification and validation features of properties that allow classifying them as receptive and / or vulnerable to infestation by phlebotomine sandflies. This project began with a selection of 145 households in the northwestern district of Belo Horizonte, ranked in three levels of receptivity/vulnerability by ACE. Then, it was selected 46 of these households, and 92 HP traps were placed for three consecutive nights in peri and intradomiciles in five evaluations. The specimens were sorted in field by the ACE and measured with laboratory technicians. Among the properties selected and confirmed as very receptive (18), 78% were positive for phlebotomine sandflies as well as the low level of receptivity (7), and receptive (14), 71% and 50% were respectively infested. It is important to note that among the positive households, three had a yield far above to the rest. Among the specimens collected in the field and classified as phlebotomine sandflies, 82% of specimens were laboratory confirmed, revealing previously a good detection by the ACE and therefore an effective training conducted in partnership with research institutions. Standardize and validate training of agents in field to identify the infested houses can enable a better planning and prioritization of vector control through risk classification of households. This work began in May 2012 and is scheduled to finish in March 2013, still leaving two evaluations that will determine the final results of the sandfly vector and other related capture. Beyond this point, other variables containing characteristics of peri and intradomiciles environments obtained by applying semi-structured questionnaire will be correlated to the abundance of the vector, as well as the association between seroprevalence and canine cases of LVH in the region.

- EPIDEMIOLOGY [1404] P 955 - CLINICAL-EPIDEMIOLOGICAL AND LABORATORY FEATURES OF PATIENTS WITH AMERICAM CUTANEOUS LEISHMANIASIS DRS-XVI, SOROCABA - SP, BRAZIL GOMES, A.H.S.1; ARMELIM, I.M.2; CORTEZ, S.V.3; MARTINS, E.P.4; BARBO, M.L.P.5; BRASIL, R.6; KANAMURA, C.T.7; IGLEZIAS, S.D.A.8; PEREIRA-CHIOCCOLA, V.L.9 1.INSTITUTO ADOLFO LUTZ, SOROCABA, SP, BRAZIL; 2.CENTRO DE LABORATÓRIO REGIONAL DE SOROCABA-INSTITUTO ADOLFO LUTZ, SOROCABA, SP, BRAZIL; 3.CENTRO DE LABORATÓRIO REGIONAL DE SOROCABA- INSTITUTO ADOLFO LUTZ, SOROCABA, SP, BRAZIL; 4.GRUPO DE VIGILANCIA EPIDEMIOLOGICA 31, SOROCABA, SP, BRAZIL; 5.PUC, SOROCABA, SP, BRAZIL; 6.CENTRO DE PATOLOGIA - INSTITUTO ADOLFO LUTZ, SÃO PAULO, SP, BRAZIL; 7.CENTRO DE PATOLOGIA- INSTITUTO ADOLFO LUTZ, SÃO PAULO, SP, BRAZIL; 8.CENTRO DE PATOLOGIA -INSTITUTO ADOLFO LUTZ, SÃO PAULO, SP, BRAZIL; 9.CENTRO DE PARASITOLOGIA E MICOLOGIA - INSTITUTO ADOLFO LUTZ, SÃO PAULO, SP, BRAZIL.

Keyword:leishmaniasis; epidemiological ; sorocaba

Abstract: The American Cutaneous Leishmaniasis (ACL) is a parasitic infectious disease that can presents different clinical aspects and epidemiological. In São Paulo state occurs a geographic expansion of this disease with changes in its epidemiological profile due to maintaining old foci or related to environmental changes caused by human activities. This study aims to demonstrate the clinical, epidemiological and laboratory features of patients with clinical suspicion of Health Care Units from 13 municipalities in the Regional Health Authority of Sorocaba. This region is the second endemic area of São Paulo’s state. This study included 56 patients, 16 female and 40 male. The observed age and number of patients vary from: 2 to 12 years (6), 13 to 23 years (9), 24 to 34 years (9), 35 to 45 years (7) and older than 45 years (25). The injury’s time observed

varied from: 1 to 5 months (37), 6 to 12 months (7), and more than 12 months in 2 patients. 11 patients did not reported lesion’s time. The lesion presence was observed in 56 patients, 31 had a single lesion, 16 had multiple injury and 9 does not mentioned. The topography of the lesions was lower limbs (28), upper limbs (12), head (2), trunk (4) and two forms with mucous nasal septum perforation. Of 56 patients studied, laboratory tests were performed: 49 (direct parasitological) whose 31 were negative and 18 were positive; 30 (PCR) with 26 positive for Leishmania (V.) braziliensis and 4 negative; 48 (Montenegro test) whose 42 were positive and 6 were negative. Histopatologic analysis was performed in 54 patients: 18 had positive results, 15 were negative and 21 were suggestive of LTA. Of the 56 patients, 26 reported that they know or have heard about the disease, 17 never heard about and 13 did not report. About the presence of forests areas nearby, 34 patients reported that live near the forest, 9 did not lived near forest areas and 13 did not informed. Of the 56 patients, 34 have pets like dogs, cats, livestock and poultry. The professions or activities developed by these patients were: farmer (7), mason (3), general service aid (5), student (8), housewife (6), mechanical (1) retired (6), merchant ( 1), carpenter (1) painter (1), chemical technician (1) and no information (15). Even today it is important to correlate the clinical, epidemiological and laboratory as criteria for case definition, understanding of transmission dynamics, action planning control, improvement and innovations in laboratory techniques and especially in the treatment of these patients.

Support: FAPESP: 2011/13939-8

- EPIDEMIOLOGY [1406] P 956 - A NEW FOCUS OF VISCERAL LEISHMANIASIS IN THE CITY OF RIO DE JANEIRO, BRAZIL PIMENTEL, J.D.1; BORGES, D.T.1; MÁSPERO, R.C.2; DA COSTA, C.M.2; FERREIRA, F.C.3; BARBOSA, P.R.A.2; DE OLIVEIRA E CRUZ, D.M.2; BEZERRA, I.O.2; GARCIA, M.H.O.2

1.REDE MUNICIPAL DE VIGILÂNCIA EM SAÚDE, SECRETARIA MUNICIPAL DE SAÚDE E DEFESA CIVIL RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 2.SUPERINTENDÊNCIA DE VIGILÂNCIA EM SAÚDE, SECRETARIA MUNICIPAL DE SAÚDE E DEFESA CIVIL RIO DE JANEIRO, RIO DE JANEIRO, RJ, BRAZIL; 3.SUPERINTENDÊNCIA DE CONTROLE DE ZOONOSES, VIGILÂNCIA E FISCALIZAÇÃO SANITÁRIA, RIO DE JANEIRO, RJ, BRAZIL.

Keyword:visceral leishmaniasis; lutzomyia longipalpis; dogs

Abstract: Introduction: Since 1977 the city of Rio de Janeiro records sporadic cases of human and canine visceral leishmaniasis (VL) in periurban areas, mainly on the slopes of the Pedra Branca and Gericinó massifs, in the western zone of the city. In June 2011 the Municipal Health Secretariat of Rio de Janeiro identified confirmed cases of VL due to Leishmania chagasi in dogs living in a shelter located in a cemetery in the north zone, near the port area of the city. An epidemiological and environmental investigation was conducted, which highlighted the presence of the sandfly Lutzomyia longipalpis and other infected dogs living near the cemetery. In January 2013 the first human case was detected. In this report, we describe, in a preliminary way, the human case and the monitoring activities of vectors and reservoirs held since the detection of this new focus of transmission, in 2011. Material and Methods: During 2012 sorological surveys were conducted to monitor the occurrence of canine VL in the area. In October 2012 a new entomological survey was made to search sandflies in the cemetery and nearby. Epidemiological surveillance activities have been strengthened through the awareness of health professionals for early detection of human cases. Information about the human case was obtained from medical records and interviews with the patient. Results: In 2012, blood samples were collected from 201 dogs and 24 (11,9%) were positive for VL in rapid test (DPP). Of these, 8 (33,3%) were positive in ELISA and 6 (75%) dogs were euthanized. We identified the presence of the sandfly Lutzomyia longipalpis at cemetery and nearby. A 29-year-old female, residing near the

cemetery, had onset of symptoms in September 2012 with fever, nausea, myalgia, vomiting and loss of appetite. Subsequently, she developed anemia, emaciation, weakness, diarrhea, jaundice, hepatomegaly, splenomegaly and edema in the lower limbs. In December 2012 she was hospitalized and in January 2013 a bone marrow aspirate was performed showing the presence of Leishmania in monocytes, macrophages and released at optical microscopy. After that, a treatment with amphotericin B was initiated. Conclusions: We report a new emerging focus of canine and human visceral leishmaniasis in an urban area of the city of Rio de Janeiro. We recommend conducting an active search for new human cases, health education activities and periodic monitoring of canine cases and vectors in this area for at least two years.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1410] P 957 - SAND FLY (DIPTERA, PSYCHODIDAE) SPECIES FROM ANALÂNDIA, DRACENA, INDAIATUBA, LIMEIRA, MONTE MOR, RIO CLARO AND VALINHOS COUNTIES, SÃO PAULO STATE, BRAZIL. CUTOLO, A.A.1; OVALLOS, F.G.2; BERNARDINETTI, U.3; PIVETTA, O.S.3; MENZ, I.4; GALATI, E.A.B.2 1.DEPTO DE VIGILÂNCIA À SAÚDE, SMS MONTE MOR, MONTE MOR, SP, BRAZIL; 2.DEPTO DE EPIDEMIOLOGIA, FACULDADE DE SAÚDE PÚBLICA, USP, SÃO PAULO, SP, BRAZIL; 3.DEPTO VIGILÂNCIA EM SAÚDE, SMS INDAIATUBA, INDAIATUBA, SP, BRAZIL; 4.DEPTO DE BIOLOGIA ANIMAL, INSTITUTO DE BIOLOGIA, UNICAMP, CAMPINAS, SP, BRAZIL.

Keyword:sand fly fauna; são paulo state; lutzomyia longipalpis

Abstract: Cutaneous (LTA) and Visceral (LVA) American Leishmaniases incidences are increasing in human and canine hosts, especially LVA, which is expanding its range through São Paulo state.

Distribution and ecology knowledge of different sand fly species is essential for leishmaniasis epidemiology vigilance. Approximately 500 hours of sand fly captures were performed using automatic light traps in rural or peri-urban areas of different São Paulo state inland counties, in random dates from 2011 to 2012, aiming identification of sand fly existing fauna. In Monte Mor county 875 specimens were captured: Nyssomyia whitmani, Migonemyia migonei, Pintomyia fischeri, Pi. monticola, Expapilatta firmatoi and Ny. neivai wich corresponded to 94.7% (829/875) of total. In Indaiatuba county 295 specimens were collected: Brumptomyia avellari, Psathyromyia aragaoi, Pi. fischeri, Pi. pessoai, Pi. monticola, Mi. migonei, Ex. firmatoi, with Lutzomyia longipalpis corresponding to 42.4% (125/295) and Ny. whitmani to 31.9% (94/295) of total. In Limeira county forest area were captured 11 specimens of the following species: Pi. monticola, Pi. pessoai, Ny. whitmani, Mi. migonei, Ex. firmatoi and B. avellari. Valinhos county forest area responded for nine specimens including Pi. fischeri, Pa. aragaoi, Pa. pascalei, Martinsmyia alphabetica and B. carvalheiroi species. Analândia county forest area had 38 specimens of Lu. longipalpis collected. Rio Claro county peri-urban zone had one specimen of Ny. neivai captured. In Dracena county urban area 1480 specimens of Lu. longipalpis and one individual of Evandromyia cortellezzii were captured. The presence of species like Ny. neivai, Ny. whitmani and Lu. longipalpis indicates risk of Leishmania transmission in some counties and demands further local studies on their ecology and transmission capability, despite of their ability in occupying anthropic environments.

- VECTOR BIOLOGY AND CONTROL: EXPERIMENTAL AND FIELD WORK [1426] P 958 - SALIVARY ANTIGEN SP32 IS THE IMMUNODOMINANT TARGET OF THE ANTIBODY RESPONSE TO PHLEBOTOMUS PAPATASI BITES IN HUMANS. MARZOUKI, S.1; ABDELADHIM, M.1; ABDESSALEM, C.B.1; OLIVEIRA, F.1; FERJANI, B.1; GILMORE, D.2; LOUZIR, H.1; VALENZUELA, J.G.2; BEN AHMED, M.3 1.LABORATORY OF TRANSMISSION, CONTROL AND IMMUNOLOGY OF INFECTION, INSTITUT PASTEUR DE TUNIS, TUNIS, TUNISIA; 2.VECTOR MOLECULAR BIOLOGY SECTION, LABORATORY OF MALARIA AND VECTOR RESEARCH, NIH, ROCKVILLE, UNITED STATES; 3.FACULTE DE ME'DECINE DE TUNIS, TUNIS, TUNISIA.

Keyword:salivary; antigen; clone

Abstract: Introduction: Zoonotic cutaneous leishmaniasis (ZCL) due to Leishmania major is highly prevalent in Tunisia and is transmitted by a hematophagous vector, Phlebotomus papatasi (P. papatasi). While probing for a blood meal, the sand fly injects saliva into the hostâ&#x20AC;&#x2122;s skin, which contains a variety of highly immunogenic compounds. We recently showed that the presence of anti-saliva antibodies was associated with an enhanced risk for leishmaniasis and identified the immunodominant salivary protein of P. papatasi as a protein of approximately 30kDa. Material and methods: We cloned and expressed in mammalian cells two salivary proteins from P. papatasi, PpSP30 and PpSP32, with predicted molecular weights close to 30kDa. We tested both recombinant proteins in serological experiments using sera from individuals living in endemic areas for ZCL and naturally exposed to P. papatasi bites. While recombinant PpSP30 (rPpSP30) was poorly recognized by sera, rPpSP32 was strongly recognized by all the tested samples. The binding of IgG antibodies to native PpSP32 was inhibited by the addition of rPpSP32.

Consistently, experiments in mice showed that PpSP32 induced the highest levels of antibodies compared to other P. papatasi salivary molecules while PpSP30 did not induce any detectable antibodies. Conclusion: We demonstrate that PpSP32 is the immunodominant target of the antibody response to P. papatasi saliva. We also show that the recombinant form of PpSP32 is similar to the native one and represents a good candidate for large scale testing of human exposure to P. papatasi bites and perhaps for assessing the risk of contracting the disease.

SATELLITE SYMPOSIUM 1 PALADIN

Title: Efficacy and safety of miltefosine in the treatment of cutaneous leishmanaisis in South America: 2013 update Abstract: This symposium will focus on the role of miltefosine in treating New World cutaneous leishmaniasis, including: an overview of the current status of cutaneous disease in the Americas; a review of the safety and efficacy of miltefosine; and the use of miltefosine in the treatment of children with cutaneous leishmaniasis in Colombia.

Speakers: Reynaldo Dietze, Núcleo de Doenças Infecciosas – UFES, Brazil Sinesio Talhari, Universidade Nilton Lins, Brasil Paulo Machado, Hospital das Clínicas, Salvador, Brazil Nancy Saravia, Corporación CIDEIM, Colombia Robert Vinson, Paladin Labs, Canada

SATELLITE SYMPOSIUM 2 DNDi

Title: DNDI-VL-2098: a nitroimidazooxazole derivative as a potential clinical candidate to treat Visceral Leishmaniasis Author names: Delphine Launaya, Denis Martina, Stéphanie Braillarda, Preeti Vishwakarmab, Suman Guptab, Sunil Purib, Vanessa Yardleyc, Mvenkata Raod, Vikram Ramanathand, H. Krishnappad, Hari Patid, Andrew Thompsone Author affiliations: a

Drugs for Neglected Diseases initiative, Geneva, Switzerland; bCentral Drug Research Institute, Lucknow, India; cLondon School of Hygiene and Tropical Medicine; dAdvinus Therapeutics, Bangalore, India; eAuckland Cancer Society Research Centre, University of Auckland, New Zealand.

Abstract text DNDi (Drugs for Neglected Diseases initiative) is a collaborative, patients’ needs-driven, not-for-profit organization whose mission is to develop new drugs for the most neglected tropical diseases, including Visceral Leishmaniasis (VL).

To this purpose, our research effort focused on the nitroimidazole chemical class, well known for its antimicrobial activity, and quickly led to the identification of DNDI-VL-2098 as a potential candidate for VL. This compound is indeed likely to fulfill DNDi’s Target Product Profile, such as: a) effectiveness against L. donovani in East Africa and India (including drug resistant strains), b) 95 % clinical efficacy, c) no contraindications, d) oral formulation, e) dosing once a day for a maximum of 10 days, f) cost under $10 per course.

DNDI-VL-2098 is a chiral nitroimidazooxazole analog prepared in 8 steps from commercially available materials. Its in vitro activity (IC50) against different strains of Leishmania donovani ranges from 0.3 to 1 µM. In VL in vivo models, DNDI-VL-2098 showed the following efficacies: -

Total cure at 12.5 mg/kg single dose or 3 mg/kg over 5 days in the acute mouse model Total cure at 50 mg/kg over 3 days or 25 mg/kg over 5 days in the chronic hamster model

The oral bioavailability of DNDI-VL-2098 varied from 70 % (rat) to 100 % (mouse, hamster). Its volume of distribution ranged from 2.0 to 2.5 L/kg (about 3-fold the total body water volume) and the mean elimination half-life ranged from 1.2 (hamster) to 6.0 h (dog).

DNDI-VL-2098 moderately inhibited the hERG channel (in transfected Chinese Hamster Ovary cells), with an IC50 of 10.5 µM. However, a telemetry study in male Beagle dogs showed no modification of QTc up to the highest dose tested (250 mg/kg). DNDI-VL-2098 was shown to be neither mutagenic nor clastogenic. In mice and rats, after single dose oral administration, the No-Observed-Effect Level (NOEL) was > 2000 mg/kg.

According to allometric scaling predictions for human PK, the minimum efficacious dose in human beings (60 kg) is expected to be in the range of 200 to 600 mg, with a once a day oral administration.

In conclusion, DNDI-VL-2098 so far shows promising properties to be developed as a clinical candidate for the treatment of visceral leishmaniasis.

Title: Towards the identification of a back-up candidate for DNDI-VL-2098 to treat visceral leishmaniasis

Author names: Delphine Launaya, Eric Chatelaina, Stéphanie Braillarda, Preeti Vishwakarmab, Suman Guptab, S.K. Purib, Vanessa Yardleyc, Louis Maesd, Andrew Thompsone Author affiliations: a

Drugs for Neglected Diseases initiative, Geneva, Switzerland; bCentral Drug Research Institute, Lucknow, India; cLondon School of Hygiene and Tropical Medicine; dLaboratory of Microbiology, Parasitology and Hygiene, University of Antwerp, Belgium; eAuckland Cancer Society Research Centre, University of Auckland, New Zealand.

Abstract text The Drugs for Neglected Diseases initiative (DNDi) is a collaborative, patients’ needs-driven, not-forprofit organization whose mission is to develop new drugs for the most neglected tropical diseases, including Visceral Leishmaniasis (VL).

Through collaboration with the TB Alliance/Auckland University, DNDi identified DNDI-VL-2098 as a potential drug candidate for VL, and is currently completing a set of preclinical studies. Since nitroimidazoles still hold great potential as treatment for neglected tropical diseases (including VL), the project team decided to focus its efforts on a back-up program. The main objectives were as follows: a) modify the core structure of DNDI-VL-2098 (nitroimidazooxazole) to encompass potential safety liabilities; b) improve pharmacological properties; c) decrease hERG inhibition potential and d) show efficacy in in vivo models.

To address these objectives, it was decided to concentrate on substituted nitroimidazooxazine analogues.

High-throughput screening of a library of nitroimidazooxazines at IPK (Institut Pasteur Korea) led to a number of hits from two subseries that were progressed into lead optimization. Medicinal chemistry work on those hit series was mainly concentrated on side chain variations, and particularly on incorporating new heteroaryl systems designed to improve the overall pharmacological profile of the current lead. To date, around 230 compounds were synthesized and screened in vitro at CSIR-CDRI against the intramacrophagic amastigote stage of the parasite (VL) expressing luciférase firefly reporter gene. From these, 190 compounds showed IC50 < 1 µM and more than 60 compounds had IC50 < 100 nM. Selected compounds with excellent in vitro potential (potency as well as microsomal stability) were evaluated in vivo. Those compounds with the most encouraging efficacies in both mouse and hamster models of VL are currently being progressed as back-up candidates. Additional studies are in progress to further characterize these and to select a back-up compound with the best chance of becoming a successful clinical candidate.

DEVELOPMENT OF NEW DRUGS FOR LEISHMANIASIS : WHAT CAN WE LEARN FROM THE IN VIVO PROFILING OF SEVERAL PROMISING CHEMICAL SERIES STEPHANIE BRAILLARD1; ERIC CHATELAIN2; CHARLES E. MOWBRAY3; DELPHINE LAUNAY4; LOUIS MAES5; ANDREW THOMPSON6.

1,2,3,4.DNDI, GENEVA - SWITZERLAND ; 5.LABORATORY OF MICROBIOLOGY, PARASITOLOGY AND HYGIENE, UNIVERSITY OF ANTWERP, ANTWERP - BELGIUM ; 6.AUCKLAND CANCER SOCIETY RESEARCH CENTRE, UNIVERSITY OF AUCKLAND, AUCKLAND - NEW ZEALAND .

Drugs for Neglected Diseases initiative (DNDi) is a collaborative, patients’ needs-driven, not-for-profit organization whose mission is to develop and make available new drugs for the most neglected tropical diseases, such as visceral leishmaniasis (VL).

DNDi’s Lead Optimization programs aim to develop and improve chemical entities that have to fulfill the Target Product Profile (TPP) in terms of efficacy, safety and cost. To this end, building a robust and accurate screening cascade for identifying potential candidates from discovery screening to preclinical development is crucial. This is a dynamic, two-way process that depends on the results obtained through in vivo profiling and, retrospectively, from clinical outcomes.

Partnership between DNDi, academic laboratories with leishmaniasis or series specific medicinal chemistry expertise and committed Contract Research Organizations led to the generation of a reasonable number of in vivo data both on pharmacokinetics and efficacy.

Efficacy data was obtained through in vivo assessment in the Balb/c mouse model, and more recently in the Syrian hamster model developed at LMPH (Laboratory of Microbiology, Parasitology and Hygiene).

This hamster model, which aims to predict results in humans as much as possible, showed robust and highly reproducible results. Supplemented by pharmacokinetic data in different species, this efficacy model allowed us to start building a preliminary assessment of the Pharmacokinetic/Pharmacodynamic (PK/PD) relationships for different new VL chemical series, such as oxaboroles and nitroimidazoles.

Moreover, evaluation of existing drugs or new formulated drugs will allow us to benchmark this model, and, more importantly, will reinforce the accuracy of DNDi’s screening cascade through a better understanding of the in vitro / in vivo profiling as a function of the different mechanisms of action.

NEW DRUGS FOR LEISHMANIASIS: FROM SCREENING TO NEW CANDIDATES FOR CLINICAL DEVELOPMENT Charles E. Mowbray Drugs for Neglected Diseases initiative (DNDi), Geneva, Switzerland

The treatment options for patients suffering from visceral leishmaniasis (VL) are limited and are often old drugs with serious limitations due to toxicity, inconvenient routes of administration, long duration of therapy and limited efficacy. There is an urgent need to deliver a new generation of safe, effective and convenient medicines to tackle this disease and combat the threat of drug resistance.

A short review of the properties and issues associated with some of the approved drugs for VL will serve to underline the on going, unmet medical need for new and improved medicines. However, there are significant challenges to targeting the protozoan Leishmania parasites which are the causative agents of VL. There are few validated anti parasitic mechanisms and furthermore the parasites are intracellular presenting additional challenges in delivering drugs to their site of action.

At DNDi we have adopted a phenotypic screening approach to identify new chemical leads. We are fortunate to have access to the screening collections of a number of generous partners; however we are faced with a number of challenges in selecting appropriate compounds to most effectively utilize our available screening capacity. Some of the lessons and conclusions from our screening campaigns will be presented.

Our screening cascade for optimizing initial hits into optimized leads suitable for pre clinical and subsequent clinical evaluation will be described and illustrated with examples of leads from novel chemical classes which present promise for future clinical studies.

Title: Treatment access challenges in leishmaniasis; recommendations Author name(s): Margriet den Boer1, Daniel Argaw1, Ivan Velez2, Jorge Alvar1,3 Author affiliation(s): 1

World Health Organization, Geneva; 2 PECET (Program for the Study and Control of Tropical Diseases), University of Antioquia, Medellin, Colombia; 3 Drugs for Neglected Diseases initiative, Geneva Abstract text In a world-wide survey performed in 2008-2010 in all countries endemic for leishmaniasis, a multitude of treatment access problems were investigated. A detailed analysis of the outcomes is the content of this presentation.

Multiple challenges for treatment access are posed by a lack of access to leishmaniasis treatment services. Medicine related factors, such as limited production, high prices, absence of pooled forecasting and procurement, and a less-than-thorough quality supervision have caused regular supply ruptures in the past with serious consequences for programs. Other factors that impede treatment access include lack of National Leishmaniasis Control Programs in the majority of the countries, or inadequate funding to roll them out. Diagnosis and treatment is not always available at primary health care level, and many leishmaniasis-endemic areas are very remote, with no health facilities and poor means of transport. Especially in coetaneous forms, patients will often use traditional medicines based on plants and caustics that are perceived as a better option than treatment offered in health care facilities.

Patients are generally extremely poor and suffer economical catastrophe when they spend time away from home in order to receive treatment, or cannot afford treatment at all. There is also a lack of awareness among patients of the serious nature of the disease, causing them to seek substandard private care. Where treatment is decentralized, inadequate trained human resources and discontinuous drug supply pose problems, and in some cases, ineffective means of diagnosis and drugs are offered, not in line with WHOâ&#x20AC;&#x2122;s recommendations.

These problems can be overcome with increased commitment of the governments of endemic countries, development partners, and international donors, and continued cooperation with pharmaceutical manufacturers to make existing treatments accessible and develop new treatments that are safe and affordable. Recently, a significant difference in treatment access has been observed in South Sudan, delivering a proper response to the recent outbreak that affected more than 25,000 patients, in which the reported case fatality rate was <5%, much lower than compared to the outbreak in the nineties (35%).

TITLE:

FROM PHASE III CLINICAL TRIALS TO IMPLEMENTATION: AN EXAMPLE OF SSG AND PM COMBINATION THERAPY IN EASTERN AFRICA

AUTHORS: Wasunna M1,2, Musa A3, Asrat H4, Khalil E3, Olobo J5, Ellis S6, Strub-Wourgaft N6, Balasegram M7 INSTITUTIONS: 1 2

Centre for Clinical Research, Kenya Medical Research Institute, Nairobi, Kenya Drugs for Neglected Diseases initiative Africa, Nairobi, Kenya

3

Institute of Endemic Diseases, University of Khartoum, Khartoum, Sudan

4

Addis Ababa University, Addis Ababa, Ethiopia

5

Makerere University, Kampala, Uganda

6

Drugs for Neglected Diseases Initiative, Geneva, Switzerland

7

MSF-Access Campaign, Geneva, Switzerland

ABSTRACT: The Leishmaniasis East Africa Platform (LEAP), founded by the Drugs for Neglected Diseases initiative (DNDi) in 2003, has three main objectives: to evaluate, validate and facilitate registration of new treatments for visceral leishmaniasis (VL) in the region; to strengthen clinical research capacity; and to serve as a base for on-going educational cooperation among Eastern African countries (Ethiopia, Kenya, Sudan and Uganda), facilitating standardization of procedures and practices in the region, within the confines of local regulations.

A phase 3 clinical trial to evaluate the efficacy and safety of paromomycin (PM) alone versus sodium stibogluconate (SSG), as well as a short course combination of SSG&PM was completed by LEAP in January 2009. Consequently, in March 2010, the World Health Organization expert working group on Leishmaniases recommended the use of SSG&PM as the first line treatment for VL in East Africa. Shortly afterwards, the Federal Government of Sudan approved this recommendation in selected Ministry of Health sites, in collaboration with DNDi and MÊdecins Sans Frontières. The process of inclusion in national treatment guidelines and registration of PM is on-going in the other LEAP member countries.

After the phase 3 clinical trials, the following steps were taken before the treatment was made available to patients: Review by the expert advisory body of national drug regulatory authorities Review by the national expert groups towards inclusion of the short course SSG&PM combination in national VL guidelines Revision of national VL guidelines and inclusion of SSG&PM on the essential medicines list in LEAP countries Registration of PM Pharmacovigilance studies were initiated Community sensitization

Training of health workers Despite these measures and political will from the LEAP country governments, challenges remain and hinder comprehensive evaluation and registration of new medicines.

Harmonization by Eastern African countries of the steps that follow successful completion of phase 3 studies is a priority to make sure effective medicines are actually available to patients.

Title: Translation from WHO Recommendation to Implementation in National Policy Author: Ellis S1, Musa A2, Burza S3, Allirol E4, 5, Lima N3, Chappuis F4, 5, Hailu A6, Khalil E2, Olobo J7, Wasunna M1, 7, Sharma B1, Balasegaram M8, Author Affiliation: 1Drugs for Neglected Diseases initiative; Institute of Endemic Diseases,Khartoum; 3 Médicos Sin Frontieras, Spain; 4Hôpital University de Genève, Geneva; Médcins sans Frontiéres, Suisse, 6 Addis Ababa University School of Medicine, Ethiopia; 7Makerere University, Kampala; 8Kenya Medical Research Institute, Nairobi; 9MSF Access Campaign, MSF Spain, KEMRI, Institute for Endemic Diseases Khartoum Abstract: The most stringent evidence usually comes from randomized controlled clinical trials that have often been conducted in centres of excellence or referral hospital settings. The results from such clinical trials are used to develop policy, which then needs to be implemented by National Control Programs in “real life settings” where most patients are treated. WHO held a technical expert committee to update the visceral leishmaniasis (VL) Guidelines in 2010 in which the treatment recommendations were revised for East Africa and South Asia. It is then up to the individual countries to implement the WHO recommended policy change. There are a number of issues that need to be addressed before recommendations based on results of phase III clinical trials can be implemented. These include registration and/or inclusion on national essential drug lists of the drugs concerned, revision and approval of national treatment guidelines, provision of training to health care workers and feasibility of giving the new treatments in resource limited setting.

Other important considerations when implementing new treatment which need be addressed include the evaluation of treatment in populations routinely excluded from clinical trials eg very young children, pregnant or lactating women. In order to address this pharmacovigilance and effectiveness monitoring of new treatments in broader population is recommended and increasingly required by regulatory authorities.

In order to address some of these issues two projects have been initiated adapted to the local situation and requirements of the individual settings to provide safety and or effectiveness data on new treatment in “real life settings”. The first project in East Africa is collecting Pharmacovigilance data on Sodium Stibogluconate and Paromomycin combination in all VL patients treated in 12 Sites in Ethiopia (3 sites), Kenya(2) , Sudan (6) and Uganda (1). A second project to evaluate safety and effectiveness of WHO recommended treatment regimens in India has started in 2 districts hospitals and 7 primary health care centres in Bihar, India. The design and status of the projects, together with some of the challenges in setting up and conducting such projects will be presented.

Title The Post Kala azar Dermal Leishmaniasis (PKDL) Consortium

Authors Ed Zijlstra a,b, Philippe Desjeux c

Authors’ affiliations a

Rotterdam Centre for Tropical Medicine, The Netherlands; b Drugs for Neglected Diseases initiative, Geneva, Switzerland; c Formerly: PATH/OWH, Divonne, France.

Abstract text Post Kala azar Dermal Leishmaniasis (PKDL) is a neglected entity within visceral leishmaniasis (VL) and occurs in up to 60% of VL cases in Sudan and 10 15% of VL in India and Bangladesh. Without control of PKDL, VL elimination efforts may not be achievable as PKDL patients are believed to be a reservoir for transmission. There are many issues that need to be addressed such as the burden of disease, pathogenesis and adequate diagnosis. Treatment is particularly difficult as in Africa most cases heal spontaneously whereas in Asia this does not seem to be the case. Current drug regimens are expensive, toxic and of long duration.In recent WHO meetings the lack of tools, gaps in knowledge and the need for a research agenda for PKDL were emphasized. Early 2012 the Institute of One World Health/ PATH and the Drugs for Neglected initiative (DNDi) launched the PKDL Consortium; in the first meeting in Delhi (June 2012) 50 experts from all endemic areas attended. The following mission statement was adopted: “The PKDL consortium is committed to promote and facilitate activities that lead to better understanding of all aspects of PKDL that are needed for improved clinical management and to achieve control of PKDL and VL.”It was agreed that the Consortium would focus on Research, Training, Advocacy and Coordination & Communication. Four Working Groups were formed to identify research priorities in Epidemiology, Diagnosis, Treatment and Pathogenesis. Cross cutting issues (between African and Asia) are given priority. In a follow up meeting in January 2013 research proposals will be developed in further detail.

The PKDL Consortium had an excellent start and will continue to act as a forum to coordinate and to stimulate all PKDL related activities.

Animal models in the evaluation and optimisation of new drugs for visceral and cutaneous Leishmaniasis Simon L. Croft, London School of Hygiene & Tropical Medicine, London WC1E 7HT, UK Animal models are a pre-requisite for drug R & D, having roles in (a) regulatory requirements, (b) as a guide to formulation / drug delivery, (c) for determination of selective activity within tissue/organ systems, (d) for determination of selective activity in acute and chronic models of infection, (e) for PK PD analysis, (f) for the evaluation of toxicity, and (g) for the identification of potential drug combinations. Appropriate models provide (i) species relevant to human infection, (ii) similarity of course of infection in animals and humans, and (iii) reproducible parasite burden. For visceral (VL) and cutaneous leishmaniasis (CL) most clinical isolates will infect rodent models (mice, hamsters, rats), parasites will infect target cells / tissues, defined parasite strains are available (sensitive and resistant), defined mouse strains available (selfcure, non-cure; species/strain specific) are available and for VL models are validated with standard drugs; CL models provide some challenges.

For drug development animal models should provide predictive data that is the efficacy and pharmacokinetics of novel anti-leishmanial compounds in acute and chronic models should predict activity in leishmaniasis patients, and the animal data should help to guide the selection of human dose levels and the design of clinical studies. Over the past decade the development of antibacterial drugs, and more recently antimalarials, the analysis of PK PD relationships has proved to be a valuable tool to improve these predictions. Information on plasma and tissue concentrations of novel compounds, on free and protein bound fractions in plasma and tissue, on the rate of kill and time course of anti-microbial effects has helped to determine exposureresponse relationships and construct PK-PD parameters that are predictive of clinical efficacy in humans. The current data available for anti-leishmanial drugs will be presented and the pathway to gaining the full data required will be discussed.

POST KALA-AZAR DERMAL LEISHMANIASIS IN INDIA â&#x20AC;&#x201C; PITFALLS IN DIAGNOSIS POONAM SALOTRA. NATIONAL INSTITUTE OF PATHOLOGY(ICMR), NEW DELHI - Ă?NDIA. Parasites of Leishmania donovani complex cause visceral leishmaniasis (VL), which is endemic in the state of Bihar, Eastern U.P. and parts of West Bengal in India. About 5 -10% cases cured of VL develop post kala-azar dermal leishmaniasis (PKDL). The need to recognize the varied presentations of PKDL and treat them as a part of VL control program has been emphasized since PKDL is a reservoir for the causative parasite. Conventional diagnostic methods based on parasite detection by stained smears, histopathology and culture have poor sensitivity (3558%) and use invasive procedures. Serological tests like DAT, ELISA and rk-39 based tests are not conclusive for PKDL diagnosis because of persistence of anti-leishmanial antibodies after VL. Such limitations have been overcome w ith highly sensitive and specific molecular tests including PCR and nested PCR. We have developed a real-time PCR assay based on kinetoplastid DNA for a rapid and reliable diagnosis of PKDL. The assay is applicable for assessment of cure and to monitor the efficacy of new antileishmanial drugs. The assay revealed that the mean parasite burden was distinctly higher (P<0.0001) in nodular lesions (n=32, 33461 parasites/ g tissue DNA) compared to papular/macular lesions (n=30, 489 parasites/ g tissue DNA). Further, Q-PCR assay using minimally-invasive slit aspirate as the diagnostic specimen offered sensitivity and specificity comparable to that w ith tissue biopsy.

Additionally, we applied Loop-mediated isothermal amplification (LAMP) technique for specific amplification of target DNA under isothermal condition, as a simplified method for parasite detection without the need of sophisticated instruments. The LAMP assay, using SYBR green for detection of amplification product, applied to PKDL samples (n=62) show ed a high sensitivity (96.8%; 95% CI: 88.9-99.1%), and specificity (98.5%; 95% CI: 92.1-99.7%). The LAMP assay could be promoted as a diagnostic tool for simpler and sensitive diagnosis of PKDL in a cost effective manner.

SATELLITE SYMPOSIUM 3, 4, 5 Bayer

Title: Genetics & immune responses in canine Leishmania infantum infection Author name(s): Drs. Christine Petersen1,2 and Lluis Ferrer3 Author affiliations: 1Iowa State University, College of Veterinary Medicine, 1600 S. Christiansen Blvd, Ames, IA, 50011, 2University of Iowa, Department of Epidemiology, College of Public Health, 105 River Street, Iowa City, IA, 52242, 3Tufts Cummings School of Veterinary Medicine, 200 Westboro Road, North Grafton, MA 01536 Abstract Symptomatic infection with Leishmania infantum in dogs has been shown to be associated with, increased blood parasite load as measured by qRT-PCR. This increased parasite load rises concomitantly with increased antibody production. Although infected dogs initially have a substantial CD4+ T cell proliferative response to L. infantum-specific antigens, as the dogs become polysymptomatic they lose this proliferative ability. This loss occurs in conjunction with decreased IFN- production and increased IL-10 production by PBMC in response to L. infantum antigen. A combination of these immunological parameters could be used to predict disease progression. The loss L. infantum-specific CD4+ T cell proliferation was correlated with increased surface expression of Programmed Death (PD)-1, an immune inhibitory receptor, leading to T cell exhaustion. Block of PD-1 ligand (B7.H1) in PBMC from asymptomatic and symptomatic VL dogs leads to regained IFN- production, antigen-specific T cell proliferation, and decreased IL-10 production. A combination of genetic and environmental factors probably explains why some dogs develop an effective immune response and others have only a partial response. A genome-wide association study of 104 boxer dogs with symptomatic infection and 115 boxer dogs with non-symptomatic infection detected regions on chromosomes 1, 4 and 20 associated with progression from infection to symptomatic disease. A single gene is not responsible for the phenotype of progression from infection to clinical leishmaniosis in the dog and it is estimated that a substantial part of the genome is affecting the trait â&#x20AC;&#x153;symptomatic infectionâ&#x20AC;? ( 1.6% of the genome). The genetic variance corresponds to 60% of the total observed phenotypic variance.

Title: Genetics & immune responses in canine Leishmania infantum infection Author name(s): Drs. Christine Petersen1,2 and Lluis Ferrer3 Author affiliations: 1Iowa State University, College of Veterinary Medicine, 1600 S. Christiansen Blvd, Ames, IA, 50011, 2University of Iowa, Department of Epidemiology, College of Public Health, 105 River Street, Iowa City, IA, 52242, 3Tufts Cummings School of Veterinary Medicine, 200 Westboro Road, North Grafton, MA 01536 Abstract Symptomatic infection with Leishmania infantum in dogs has been shown to be associated with, increased blood parasite load as measured by qRT-PCR. This increased parasite load rises concomitantly with increased antibody production. Although infected dogs initially have a substantial CD4+ T cell proliferative response to L. infantum-specific antigens, as the dogs become polysymptomatic they lose this proliferative ability. This loss occurs in conjunction with decreased IFN- production and increased IL-10 production by PBMC in response to L. infantum antigen. A combination of these immunological parameters could be used to predict disease progression. The loss L. infantum-specific CD4+ T cell proliferation was correlated with increased surface expression of Programmed Death (PD)-1, an immune inhibitory receptor, leading to T cell exhaustion. Block of PD-1 ligand (B7.H1) in PBMC from asymptomatic and symptomatic VL dogs leads to regained IFN- production, antigen-specific T cell proliferation, and decreased IL-10 production. A combination of genetic and environmental factors probably explains why some dogs develop an effective immune response and others have only a partial response. A genome-wide association study of 104 boxer dogs with symptomatic infection and 115 boxer dogs with non-symptomatic infection detected regions on chromosomes 1, 4 and 20 associated with

progression from infection to symptomatic disease. A single gene is not responsible for the phenotype of progression from infection to clinical leishmaniosis in the dog and it is estimated that a substantial part of the genome is affecting the trait “symptomatic infection” ( 1.6% of the genome). The genetic variance corresponds to 60% of the total observed phenotypic variance.

Title: Update on the pathomechanisms of clinical canine leishmaniosis Author name(s): Alexander F. Koutinas Author Affiliations: Prof (ret) of Companion Animal Medicine (Thessaloniki), Volos, Greece Dogs with overt leishmaniosis (CanL) show a Th2-like immune response resulting in tissue damage via different pathomechanisms, notably lymphoplasmocytic, lymphohistiocytic and/or granulomatous inflammation (e.g. nodular dermatitis), immune complex deposition (e.g. polyarthritis, glomerulonephritis, uveitis) and auto-antibody production (e.g. thrombocytopenia, myositis). Although the Th-cell dichotomy scenarios, presumably imposing on the immunopathogenicity of CanL, are complex and still unclear, Leishmania infantum infection in fact appears to induce a mixed Th1/Th2 response in either asymptomatic or sick animals. Macrophages play a central role, through the secretion of IFN- , TNF- and IL-2 cytokines by activated T-cells and nitric oxide production by themselves, hence mediating the intracellular killing of Leishmania amastigotes by apoptotic cell death. Macrophages are also lysed by CD8+ and/or CD4+ cytotoxic cells in a histocompatibility complex-restricted process that is suppressed in the symptomatic dog, only to compensate with an exaggerated IgG response to the parasite, that is the cornerstone of the underlying pathology and clinical disease. For some forms of the latter, IL-1, IFN- , TNF- , and acute phase proteins have been incriminated. The clinical disease is basically expressed as exfoliative and/or ulcerative dermatitis, with or without nasodigital hyperkeratosis and onychogryphosis, glomerulonephritis, temporal muscle atrophic myositis, uveitis, keratoconjuctivitis sicca, epistaxis and/or polyarthritis. Chronic colitis, hepatitis, vasculitis, myocarditis, osteomyelitis, and other conditions may appear with the clinical mainstay of CanL. An interplay between immune complex deposition/complement activation and granulomatous inflammation is most likely responsible for their induction. The pathomechanisms of the disease involve epistaxis (thrombocytopathy, serum hyperviscosity, rhinitis), myositis, exfoliative dermatitis, ulcerative dermatitis (role of vasculitis or cryoglobulinemia), cutaneous nodules (excessive granulation tissue formation) and keratoconjuctivitis sicca (periductal reaction & secretory retention, lacrimal adenitis & decreased tear production). Finally, the clinicopathological cadre of CanL contains anemia, hyperglobulinemia and profuse proteinuria, that may be asymptomatic, or associated with chronic kidney disease and/or nephrotic syndrome and may lead to thromboembolic disease or contribute to the dramatic deterioration of body condition.

Title Update on vectorial and non-sand fly transmission of Leishmania infantum Author name(s): Filipe Dantas-Torresa,b Author Affiliations: a Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, 50670-420 Recife, Pernambuco, Brazil. bDipartimento di Medicina Veterinaria, Università degli Studi di Bari, 70010 Valenzano, Bari, Italy

Abstract text Sand flies (subfamily Phlebotominae) are the principal vectors of Leishmania parasites, though biting midges may be vectors among marsupials in northern Australia. Of over 900 sand fly species, less than 100 have been regarded as proven or putative vectors of Leishmania parasites.

With the widespread use of polymerase chain reaction-based methodologies, several sand fly species have been found to carry genetic material of Leishmania spp., which suggests that the list of potential vectors of Leishmania parasites may be larger than currently appreciated. For instance, 21 sand fly species have been regarded as proven or putative vectors of Leishmania infantum in the Old World, whereas only 8 species in the Americas, according to a recent review. Considering the diversity of sand flies found in the New World, further research might reveal the existence of other putative vectors. While primarily transmitted by sand flies, L. infantum transmission may also occur by other ways, such as venereal transmission, congenital transmission, blood transfusion, and through needle sharing. The existence of secondary routes of L. infantum transmission in dogs (e.g., congenital, transfusional and venereal transmission) has been demonstrated experimentally and under natural conditions. There has also been speculation about the role of fleas and ticks in the transmission of L. infantum between dogs, but no convincing scientific evidence has been produced so far. Interestingly, the emergence of canine leishmaniasis among Foxhounds in the United States has been associated to non-vectorborne transmission, but the existence of a sand fly vector (e.g., Lutzomyia shannoni and Lutzomyia vexator) in this country cannot be ruled out. Further research is needed to assess the vectorial competence of different sand flies currently referred to as putative vectors in both Old and New Worlds. It is also fundamental to understand the epidemiological significance (if any) of secondary modes of transmissions in the maintenance of L. infantum in an area where competent sand fly vectors are absent, although it seems unlikely that this parasite could subsist for long periods in a dog population by means of these routes.

Title Epidemiology of Canine Leishmaniosis in Mediterranean Basin Author name(s): Patrick Bourdeau Author Affiliations: Unit of Parasitology Dermatology Mycology. Campus National Veterinary School of Oniris. University of Nantes, Atlanpole, 44307 Nantes, France The understanding of the epidemiology of canine leishmaniosis (CanL) (Leishmania infantum typically Li. MON1 in dogs with Phlebotomus perniciosus as common vector) in the Mediterranean basin has recently progressed. In the northern part (Portugal to Greece), several other strains have been isolated from dogs (i.e.: MON11 27 77 105 108 199). The endemic zones (high % of positive dogs by molecular/serological diagnostic tools) are linked to abundant vectors including P. ariasi (west), P. neglectus (east) and P. tobbi, P. perfiliewi, P. longicuspis. It is now clear that Leishmania may also develop in wild or domestic mammal species, with possible role as reservoirs. Periendemic zones are surrounding active areas with expanding infection in dogs in south western France, northern Italy, and northern Spain (increase of vector populations, or travelling dogs). Non endemic (cryptodemic) areas are characterized by CanL cases where sandfly transmission is unlikely; the non vectorial transmission (venereal, vertical, direct) is now accepted with potential significance when autochthonous cases are described. Good health management, rapid detection, specific treatment and preventative methods influence the evolution of infection both at individual level and collective level (reduction of the infectivity to vector). The cases of human visceral leishmaniasis (VL) are now mainly observed in severely immunocompromised patients whereas a significant % of healthy population is detected

positive. In the eastern (Turkey to Egypt) and north African (Egypt to Morocco) Mediterranean, additional strains and suspected vectors are present (MON: 27–77–98–108; P. tobbi, P. syriacus, P. kandelakii and MON: 34 37; P. ariasi, P. perfiliewi, P. longicuspis, P. langeroni). The situation is possibly complicated by the overlap with other zoonotic Leishmania spp. that focally infect dogs (i.e.: L. tropica).

In these areas the epidemiology is influenced by arid climate and abundance of stray dogs (growing but still limited population of clinically healthy dogs) and the number of VL cases remains important. In many European countries human cases decreased spontaneously (no organized control of infected dogs or vectors), and infection is well controlled in dogs at early stages by treatment and preventative methods (prevent infection and/or role of source). This illustrates the importance of the concept of improvement of both human and canine health conditions (One Health approach).

Title Epidemiology of canine leishmaniasis in South America Author name(s): Filipe Dantas-Torresa,b Author Affiliations: a Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães, 50670-420 Recife, Pernambuco, Brazil. bDipartimento di Medicina Veterinaria, Università degli Studi di Bari, 70010 Valenzano, Bari, Italy Abstract text Canine leishmaniasis (CanL) is widespread in South America, where dogs may be infected by different species of Leishmania (e.g. L. amazonensis, L. braziliensis, L. infantum, L. panamensis and L. peruviana). Several phlebotomine sand flies have been regarded as proven or putative vectors of Leishmania parasites in South America, which further complicates the epidemiological puzzle of CanL in this region. CanL is prevalent in several South American countries and territories, and is spreading in southern Brazil, Argentina and Paraguay. The recent finding of Lutzomyia longipalpis in northern Uruguay indicates the potential risk for the introduction of L. infantum (if not yet introduced) in this region. Similarly, the recent report of L. braziliensis infection in a cat from French Guiana indicates that this parasite may also be circulating among dogs in this region. Studies suggest that the level of exposure to Leishmania spp. in endemic areas of South America may be dependent on the sex, age and breed of the dogs, varying from region to region, according to dog population and criteria of exposure (e.g. diagnostic tools used). For example, some studies reported higher prevalence rates of antiLeishmania antibodies among males in comparison with females, whereas others found no association between dog gender and seropositivity. Similarly, some authors found a high seroprevalence in young dogs (<1 y) whereas others in older dogs (1–6 y). Furthermore, some authors have reported a higher seropositivity in some breeds (e.g. Boxer and Cocker) in comparison with mongrel dogs. Low family income, owner’s knowledge regarding the vector, the dog staying predominantly in the backyard, and a lack of previous serological examination have been associated to the risk of L. infantum infection dogs in south-eastern Brazil. In northeastern Brazil, dwellings with a history of dogs culled by public health authorities in the last 12 months had higher risk of having at least one seropositive dog as compared with dwellings with no history of dog culling. Interesting, another study in north-eastern Brazil indicated that higher owner education, pet attachment and perception of risk/disease were associated with decreased levels of dog seropositivity and with willingness to purchase preventatives for their pets. Further research is needed to piece together the puzzle surrounding the epidemiology of CanL in South America, which appears to be driven by several biotic and abiotic factors.

Title New findings on old and emerging reservoir hosts of Leishmania infantum Author name(s): Gad Banetha Author Affiliations: a School of Veterinary Medicine, Hebrew Univeristy, P.O. Box 12, Rehovot 76100, Israel Abstract text Leishmaniasis caused by Leishmania infantum, or its synonym Leishmania chagasi, is a major potentially fatal zoonotic infection. The disease involves mainly humans and dogs, who are considered the main peridomestic reservoir host for human infection, and also suffer from severe clinical disease. In addition, natural L. infantum infection has been recorded in cats, horses, pigs and in a variety of wild mammal and marsupial species including: golden jackals (Canis aureus), red foxes (Vulpes vulpes), gray wolves (Canis lupus), crab-eating foxes (Cerdocyon thous), opossums (Didelphis albiventris and D. marsupialis), lynx (Lynx pradinus), hares (Lepus granatensis & L. yarkandensis), badgers (Meles meles), mongooses (Herpestes ichneumon), pine martens (Martes martes), genets (Geneta geneta),weasels (Mustela nivalis), seal (Monachus monachus), bat (Carollia perspicillata), rats (Rattus rattus & R. norvegicus) and other rodent species. Not every infected animal species may serve as a reservoir host, e.g. be responsible for parasite transmission to humans or animals. Reservoir hosts for Leishmania should have several characteristics in order to serve as efficient sources of blood meals with parasites for sand fly vectors: they should be abundant, infected at a high rate, attractive and infectious to sand flies, and able to maintain infection year round. The identification of blood meal belonging to a certain animal host in a considerable number of naturally-infected sand flies in a survey of an endemic region may provide evidence for the potential of this host to serve as reservoir. Apart of dogs and humans, only a small number of animal host species have been studied for their capacity to infect sand flies with L. infantum by xenodiagnosis. Studies have shown that domestic cats, hares, black rats, opossums and crabeating foxes can infect sand flies under experimental conditions. However, the mere ability to infect sand flies does not imply that the host is an epidemiologically-important reservoir which plays a role in sylvatic or domestic transmission at the population level. The transmission of L. infatnum from domestic dogs undoubtedly constitutes the main route for human infection in most endemic areas globally. In these areas people share the same habitat with dogs infected for long durations. Although asymptomatic dogs are infectious to sand flies, dogs with clinical signs of disease have been shown to have high parasite loads and be more infectious.

Title: "Emergence of Human and Canine Visceral Leishmaniosis in Argentina" Author name(s): José Octavio Estévez Author affiliation(s): Veterinaria del Oeste, Av. Lavalle 2574, Posadas, Misiones, República Argentina Abstract text Since April 2006 when the first cases of Human and Canine Visceral Leishmaniosis (VL) were detected, Argentina became part of the list of South American countries affected by this disease. Unlike what happened in Brazil where the disease expanded gradually from the rural environment of the North East to the big urban areas. Argentina’s first outbreak occurred in the city of Posadas which has almost 400.000 inhabitants. The population of Posadas is heavily

exposed to mass media and dog owners are committed to their animals as in other modern societies. In this context, the emergence of the disease triggered conflicts between dog owners and the city authorities, mainly because euthanasia of dogs with VL was the main measure of control suggested by the authorities.

Nowadays, VL has expanded throughout the Northeast Region of Argentina and the presence of Lutzomyia longipalpis has been detected in five provinces (Misiones, Corrientes, Chaco, Formosa, and Entre Ríos), with a descending gradient of canine cases from north to south.

Most of the cases are in the province of Misiones, in the extreme Northeast of the country, between Paraguay and Brazil. Other cases have been reported in geographically separate areas, such as the northwest of the country (Santiago del Estero), where the presence of L. longipalpis was not detected, and where Lutzomyia migonei could be involved.

The total number of humans affected by VL between 2006 to 2012 is 103 with 11 deaths. About 80% of the patients are from Misiones province. The others are from the provinces of Corrientes and Santiago del Estero where phlebotomines are present, or from distant areas due to the frequent movement of people and animals. The information available about dogs is still poor and there is no reliable data about prevalence in different cities with the exception of certain places.

In the city of Posadas where the disease was first diagnosed, some official information indicated that the prevalence of dog infection is about 30% and this data is confirmed also from private practices.

The experience of veterinarians who treat affected dogs shows that there is a great compliance of owners who receive adequate information and are likely to carry out the treatment, monitoring of dog progression and topical vector control.

Despite this, the official policy in Argentina is to favour the elimination of positive animals and discourage treatment and vaccination against the disease.

Title: Feline leishmaniosis: top ten frequently asked questions (FAQ) Author names: Maria Grazia Pennisia,c and Laia Solano-Gallegob,c Author affiliations: a

Dipartimento di Scienze Veterinarie, Università di Messina, Italia;bDepartament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain; c The authors are members of the LeishVet association.

Abstract text: In the last years, feline leishmaniosis has mostly received attention from an epidemiological point of view but many questions remain to be answered. The ten most frequently asked questions are: 1) Which Leishmania species are responsible for the feline infection and which is their geographical distribution?

Molecular characterization has confirmed that the cat is infected by the same species found in dogs or humans and Leishmania infantum is the species responsible for the feline infection in the Mediterranean area. 2) What is the prevalence of L. infantum infection in cats in endemic areas? Studies on prevalence performed in these areas provided a wide range of positivity: 0-68.5% for IFAT (cut off range 1:21:100) and 0-61% for blood PCR. 3) How does transmission of Leishmania occur in the cat?

Phlebotomine sand flies are involved because they feed on cats and can become infected after feeding on a cat affected by feline leishmaniosis. 4) Does L. infantum infection cause clinical manifestations in cats?

Case reports published in the last 25 years show that the most frequent findings are skin lesions (mainly ulcerative or nodular dermatitis) and lymph node enlargement but other abnormalities can also be found.

Hyperglobulinemia is often seen. 5) Are there other conditions associated with the disease or L. infantum infection? The most common association is the occurrence of Feline Immunodeficiency Virus antibodies sometimes detected also with Feline Leukemia Virus antigenemia. 6) How is feline leishmaniosis diagnosed? Diagnosis is confirmed by direct methods (cytology, histology, culture, PCR) or serology (IFAT, ELISA). 7) How is feline leishmaniosis caused by L. infantum treated? Allopurinol is the most frequently used drug. 8) What is the expected clinical response to treatment? clinical improvement is usually obtained with a long term course of therapy. 9) Can Leishmania infection be prevented in cats? Currently proven preventative measures such as specific topical insecticides have not been studied in cats. 10) When cats should be tested for leishmaniosis? Cats living or travelling in endemic areas should be tested in case of clinical manifestations compatible with leishmaniosis. In conclusion, information on feline leishmaniosis is growing but further studies are needed to investigate gaps of knowledge that exists mainly on the pathogenic role of Leishmania spp. in cats and on the actual role of infected cats as reservoir in this infection.

Title: Update on the diagnosis of canine leishmaniosis in the Old World

Author name(s): Luís Cardosoa,c, Laia Solano Gallegob.c

Author affiliation(s): a

Department of Veterinary Sciences, School of Agrarian and Veterinary Sciences, University of Trás os Montes e Alto Douro, Vila Real, Portugal; bDepartament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Spain; cThe authors are members of the LeishVet association.

Abstract text: The purposes for which the diagnosis of Leishmania infantum infection is carried out include: 1) to confirm clinical illness; and 2) to investigate the presence of “infection” for epidemiological studies, for screening clinically healthy dogs living in endemic regions, to prevent transmission from subclinical carriers by blood transfusion, to avoid the importation of infected dogs to non endemic countries, and to monitor response to treatment. For these reasons, it is important to separate Leishmania infection from disease and to apply different diagnostic techniques accordingly. Accurate diagnosis of canine leishmaniosis (CanL) often requires an integrated approach consisting of a clinicopathological assessment and specific laboratory tests. Clinical history, a thorough physical examination and several routine diagnostic tests such as complete blood count, biochemical profile, urinalysis and serum electrophoresis can help to raise the suspicion index for this disease. The detection of L. infantum infection in dogs includes parasitological (cytology, histology, immunochemistry and culture), molecular (conventional, nested and real time polymerase chain reaction [PCR]) and serological methods (qualitative and quantitative antibody tests). Several different assays with various target sequences using genomic or kinetoplast DNA have been developed and various tissues (including blood, lymph

node, bone marrow, spleen, skin and conjunctival swab) can be used for PCR detection of the parasite in dogs. Real time PCR allows the quantification of Leishmania loads in tissues of infected dogs, which is important for diagnosis as well as for follow up during treatment. Detection of specific serum antibodies should preferably be based on quantitative serological techniques, such as the immunofluorescence antibody test (IFAT) and enzyme linked immunosorbent assay (ELISA). High antibody levels are associated with severe parasitism and disease. However, the presence of lower antibody levels is not necessarily indicative of disease and further work up is necessary to confirm CanL by other diagnostic methods such as cytology, histopathology and PCR. The challenges of serology include cross reactivity with other pathogens and antibodies elicited by vaccination against CanL. Information provided by PCR should not be separated from the data obtained from clinicopathological and serological evaluations. These should all be combined together for a comprehensive assessment.

Title: Update of the Diagnosis of Canine Leishmaniosis in the New World – Brazilian perspectives Author name(s): Vitor Márcio Ribeiro Author affiliation(s): School of Veterinary MedicinE, Catholic University, Rua do Rosário, 1081, Bairro Angola, CEP 32604115, Betim – Minas Gerais, Brazil vitor@pucminas.br Abstract text The diagnosis of canine leishmaniasis (CL) cannot be based on the physical examination, because 50% of cases may be asymptomatic. It needs to be based on physical, serological, parasitological and routine exams including complete blood count, biochemistry and urinalysis (Solano-Gallego et al., 2011). There are no laboratory techniques that can provide 100% sensitivity and specificity. In Brazil, serological, molecular and parasitological techniques are used. The serological assays used are ELISA and indirect immunofluorescence (IFI) with an IFI cut-off of 1:40, as directed by the Ministry of Health (MoH).

However, studies have shown that IFI has variable sensitivity (72-100%) and specificity (52-100%) (Romero & Boelaert, 2010) as well as ELISA (30-100%) and (64-100%), respectively (Ferreira et al., 2007; Porrozzi et al., 2007; Figueiredo et al., 2010). Since the Brazilian MoH determines the elimination of positive dogs, thousands are killed with false positives diagnoses (Alves & Bevilacqua, 2004). The scientific and civil communities question the ethical stance of the MoH, because examinations with low dilution are indicated only for screening (Campino & Maia, 2008). The recommended IFI titres for clinical diagnosis is 1:160 (Ribeiro et al., 2013). An IFI result with low titre may indicate prior exposure. IFI levels 1:160 can already be associated with clinical disease and increased parasite load (Reis et al., 2006; Baneth & Aroch, 2008). A rapid test is being implemented in Brazil, using the recombinant antigen K28 (rK26 and rK39) of Leishmania infantum. This test is used for screening, and positive samples are subjected to ELISA for confirmation. Its sensitivity is 47-98% and specificity 6896% (Schubach, 2011; Grimaldi et al., 2012). It detects early infections reducing the risk of false positives. Inconclusive serologic tests must be complemented by a parasitological exam. Cytology of the bone marrow has a sensitivity of 40-75%. Immunohistochemistry is used to evaluate the infectivity of dogs before and during treatment (Ribeiro, 2007). Molecular techniques are used in individual cases.

They have high sensitivity and specificity and enable the quantification of the parasite load by real time PCR. This is important for monitoring treated dogs. The clinician should select the exams needed for diagnosis and use routine testing to assess the animal state, select treatment and establish the prognosis.

Title New perspectives on canine leishmaniasis I therapy: parasiticidal treatments Author name(s): Guadalupe Miró Author Affiliations: Animal Health Department. Veterinary Faculty. Universidad Complutense de Madrid. Spain

Abstract text Apart from its zoonotic nature, canine leishmaniasis (CanL) continues to be of major importance in veterinary medicine from the clinical standpoint, since there are several clinical conditions in dogs that can be very serious if proper treatment is not provided.

Before deciding which treatment to use on a dog with leishmaniosis, it is necessary to evaluate the clinical signs, clinicopathologic parameters and the results of a quantitative serological test. With these three values, the LeishVet Group designed a four-stage clinical staging system. Depending on the clinical staging, different treatment regimens are recommended and the prognosis can be estimated.

The treatment of this immune-mediated parasitic disease includes applying drugs with leishmanicidal or leishmanistatic activity, as well as those whose active principle has immunomodulating effects.

To date, an active principle which is 100% effective has not been found since, generally speaking, most dogs improve from a clinical or pathological point of view but a “parasitological cure" is not achieved. In many cases, relapses occur after a period of time which depends on the drugs used and the individual immune response of each patient.

Pentavalent antimonials and miltefosine still remain the drugs of choice for the treatment of CanL, as they usually induce clinical remission, and partial reduction on the parasitic burden although they do not prevent relapses; for this reason they are normally combined with allopurinol.

Other drugs, such aminosidine, furazolidone, marbofloxacine, perifosine, OIPC, have been also recently evaluated, either alone or combined, with mixed results.

We will review, update and assess the most recent clinical trials conducted to evaluate these new treatments to advance the current understanding of CanL parasiticide therapy.

Title: New perspectives on canine leishmaniasis therapy II: immune potentiating treatments Author name(s): Lluis Ferrer Author affiliation(s): Department of Clinical Sciences, Cummings School of Veterinary Medicine, Tufts University, Massachusetts, USA Abstract text Clinical canine leishmaniosis is both an infection and an immunodeficiency. Dogs with clinical leishmaniosis develop a type of immune response that is unable to control the progression of the infection and the development of lesions and clinical signs. Once the disease has developed the animals show signs of immune-deficiency and immunopathologic abnormalities. Numerous attempts have been made to help the immune system to control the infection (immunotherapy), including:

(1) Non-specific immune potentiating drugs. The only one with evidence of efficacy is domperidone, marketed in some European countries to prevent leishmaniosis and to treat mild/early disease. Domperidone is a prolactinogogue drug which induces an increase of the T cell responses and of the phagocytic function of macrophages and neutrophils. Controlled trials have demonstrated that is a safe and effective alternative for the treatment of early/mild cases and seropositive animals. TLR activators (imidazoquinolines: imiquimod, resiquimod) are clearly helpful in cutaneous leishmaniosis and are promising in visceral leishmaniosis.

(2) Cytokines. There are a few experimental trials using -IFN, IL-12, anti-IL-10 and an IL-10 receptor antagonist, but the results have been only partially satisfactory or inconsistent. These treatments are very expensive and are not available for use in clinical cases. Only canine -IFN has been marketed so far (in Japan).

(3) Vaccines. There are several studies and trials demonstrating that vaccines can be used as therapeutic drugs in dogs with clinical leishmaniosis. In a clinical trial in Brasil the Leishmune® vaccine reduced the clinical signs and the parasite load, modulating the outcome of the infection and the dog's potential infectivity to phlebotomines. In another trial, the subunit vaccine Leish111f + MPL-SE was effective in the treatment of dogs with mild disease but not of dogs with severe clinical leishmaniosis. In Spain, an auto-vaccine prepared with parasites isolated/cultivated from the ill dog is marketed, although no clinical trials showing efficacy have been published.

Immunotherapy in canine leishmaniosis is certainly challenging, but has clearly some advantages that make it very attractive. Less and milder side effects than traditional chemotherapy, absence of resistance and the possibility of using it in combined protocols together with parasiticidal drugs, are some of them.

Title: Vaccination against canine leishmaniasis in Brazil

Author names: Clarisa Beatriz Palatnik de Sousa

Author affiliation: Laboratório de Biologia e Bioquímica de Leishmania, Instituto de Microbiologia “Paulo de Góes”, CP 68040, 21941-902. Universidade Federal do Rio de Janeiro, (UFRJ), Rio de Janeiro. Brazil, immgcpa@micro.ufrj.br

Abstract text: Three different veterinary vaccines were licensed for prophylaxis against canine zoonotic visceral leishmaniasis (ZVL): Leishmune® (2004) and Leishtec® (2007) in Brazil, and CaniLeish® (LiEsap) in Europe (2011). Leishmune® and CaniLeish® were licensed after the demonstration of their vaccine efficacies in kennel and field assays while Leishtec® was licensed based on the results of one kennel assay only. Leishmune®, composed of the Fucose–Mannose ligand antigen of L. donovani promastigotes and the QS21 and deacylated saponins of Q. saponaria was immunogenic and immunoprophylactic in mice and hamsters and in dog field trials. After two years of vaccination, only 1% of 550 vaccinated and exposed dogs died of ZVL and 1.2% was symptomatic while 39% of deaths and 20.6% of symptomatic cases were detected among untreated exposed controls. The vaccine reduced the parasite burden accessible for sand flies. The antibodies block the transmission of the disease. It promoted parasitological and clinical cure of dogs while sterile cure required immunochemotherapy. Leishmune® induced enhanced levels of IFN , NO and anti L. chagasi IgG2, the early and persistent activation of

neutrophils and monocytes, increased the CD8+ T cells expressing IFN and diminished the CD4+/CD25+ T cell counts. Vaccinated dogs were not seroreactive and the vaccine displayed an additive effect over dog culling, promoting decrease of the incidence of canine & human ZVL in endemic areas.

Its main antigen, the Nucleoside hydrolase (NH36) hydrolizes nucleosides and releases purines for the parasite DNA replication, is a target for anti leishmanial drugs. Protection against L. chagasi in mice is related to its C terminal domain (F3= amino acids 199 314) and is mediated mainly by a CD4+ T cell driven response with a lower contribution of CD8 + T cells.

The Leishtec® vaccine, on the other hand is composed of the A2 recombinant amastigote antigen and Q. saponaria saponin. The A2 antigen contains an immunogenic epitope for CD4+ T helper cells and multiple repetitive units encoding CD8+ T lymphocyte (CTL) epitopes. The A2 antigen is associated to the ability of viscerotropic species to migrate to visceral organs. In different formulations, Leishtec® showed protection in mice and in a kennel assay in Beagles (seven vaccinated dogs and four controls).

The Leisthec® vaccine was tested in a pre clinical trial in Rhesus monkeys and in a dog field assay for prophylaxis but the results are still unpublished.

Support: FAPERJ; CNPQ

Title: Current status of vaccination against canine leishmaniosis in Europe Author name: Gaetano Oliva Author affiliations: Department of Veterinary Clinical Sciences, University Federico II, Naples, Italy.

Canine leishmaniosis (CanL) is endemic in several European Countries, particularly in those of the Mediterranean Basin area. Current climatic and ecological changes have also permitted some sandfly species to colonize some continental European areas that till a few years ago were considered Leishmania-free. CanL spread is also possible because of the absence of strict limits on the importation of infected dogs from endemic to non-endemic countries. For these reasons, CanL constitutes a challenge for veterinary and public health in most parts of Europe. The main strategies for control of Leishmania infection are the use of repellents against sandflies and the very recent introduction of a specific licensed CanL vaccine, (Canileish®, Virbac). This product consists of cultured Leishmania infantum excreted secreted proteins (ESP), the dominant antigen of which is the promastigote surface antigen (PSA).

Canileish® is adjuvanted with purified Quillaja saponaria extract (QA-21) which is demonstrated to induce a good cell-mediated response. The vaccine is indicated for the active immunization of Leishmania-negative dogs from 6 months of age and induces a 1-year duration of immunity. To date there are few published studies about CaniLeish® efficacy. However it seems clear that this vaccine produces a strong T helper1 (Th1)-profile cell-mediated response within three weeks of completing the primary course, and that this response reduces the parasite load in pre-infected macrophages in vitro.

Moreover, a two year field trial performed on 90 beagle dogs in Italy and Spain showed that although the vaccine did not protect dogs from initial infection, it decreased the risk of developing progressive infections by about four fold in natural conditions of high intensity of transmission. More than 1 million vaccine doses have been sold in Europe, mostly in Portugal, Spain, Italy, France and Greece, corresponding to 2% of the well-cared for canine population (15 millions). To be applied on a large scale the most relevant points that need clarification are the capability of the vaccine to protect dogs for long periods independently from the endemicity of the area, the difficulty to distinguish vaccinated from infected dogs by serological methods and the infectivity of vaccinated dogs to phlebotomine vectors.

Vaccination should be regarded as one of multiple strategies (including control of sandfly exposure and control of stray dogs) in the management of this disease within the dog population.

Title Update on insecticides for the prevention of canine leishmaniosis Author name: Domenico Otranto Author Affiliations: Dipartimento di Medicina Veterinaria, UniversitĂ degli Studi di Bari, 70010 Valenzano, Bari, Italy Abstract text (word count: 2473) Canine leishmaniosis (CanL) by Leishmania infantum is worldwide diffused and endemic in southern Europe, Africa, Middle and Far Eastern countries and in the Americas. In southern Europe, where CanL is endemic, about 4,000 new human cases are reported yearly. Human and canine infections are closely associated and they typically occur in â&#x20AC;&#x2DC;hotspotsâ&#x20AC;&#x2122; of infection where environmental and climatic conditions are suitable for the development of the vectors. Over the last decades, considerable efforts have been directed towards developing novel and cost-effective control strategies against this disease in endemic areas. However, due to the complexities involved in the transmission of L. infantum, the control of CanL has proven challenging, and none of the proposed strategies (e.g., dog culling in Brazil) have yielded satisfactory results so far. For example, the use of insecticide house spraying against phlebotomine sand flies has been unsuccessful in Brazil, due to the fact that vectors are present during the whole year. Concerns regarding potential environmental and human health hazards have prompted the substitution of organochlorines by synthetic pyrethroids. The use of pyrethroids with repellent properties in spot-on formulations or as impregnated collars shown to be useful and cost-effective to reduce the risk of L. infantum infection in dog populations. For example the use of deltamethrinimpregnated dog collars resulted in a protection rate against CanL from 50% to 86%, over two consecutive transmission seasons in Europe. Recently, a study investigated the efficacy of a collar containing a combination of 10% imidacloprid and 4.5% flumethrin to prevent L. infantum infections in young dogs from a hyper-endemic area of southern Italy. Sixty-three dogs were collared (Group A) while 61 were left untreated (Group B), from March-April 2011 until March 2012. Serological, cytological and molecular tests were performed to detect the presence of L. infantum in multiple tissues. At the end of the trial, no dog from Group A proved positive for L. infantum at any follow-up, whereas 22 dogs from Group B were infected (incidence density rate = 45.1%). Results indicated that collars conferred longterm protection against L. infantum infection in dogs, representing a reliable and sustainable strategy to control the spread of CanL in hyper-endemic areas. This strategy will also ultimately result in the reduction of associated risks to human health.

SATELLITE SYMPOSIUM 6 SANOFI

MOVING FORWARD IN THE MANAGEMENT OF CUTANEOUS LEISHMANIASIS Co Chairs:

Ana Rabello Fiocruz, Belo Horizonte, Brazil Robert Sebbag Access to Medicines, Sanofi, Paris, France

Improving access to leishmaniasis treatment Robert Sebbag Access to Medicines, Sanofi, Paris, France Public private partnerships in leishmaniasis : the WHO perspective Daniel Argaw WHO, Geneva, Switzerland Development of a multidisciplinary model of care for CL patients in Pernambuco Otamires Alves da Silva Centro de Pesquisas Aggeu Magalhaes, Fiocruz, Brazil A retrospective cohort analysis of the efficacy and safety of meglumine antimoniate and SSG in Peruvian patients with CL Alejandro Llanos Cuentas Alexander von Humboldt Institute, Lima, Peru Development of a topical meglumine antimoniate formulation: 1st in vivo results Bartira Rossi Bergmann Instituto de Biofísica Carlos Chagas Filho, Rio de Janeiro, Brazil Standardised criteria for clinical trials in CL Byron Arana DNDi, Geneva, Switzerland “Moving forward in the management of cutaneous leishmaniasis"

Each year, according to WHO estimates (PLOS One 7, May 2012, e 3671) between 700,000 and 1. 2 million patients will be affected by cutaneous leishmaniasis (CL), divided fairly evenly, between the 3 principal endemic foci, the Americas, the Mediterranean Basin, and Western Asia, from the Middle East to Central Asia. As is recognized by all, there is a need in the management of CL for greater access to treatment for all affected patients, and for new treatments that are better adapted to patients’ needs.

This symposium, chaired by Prof Ana Rabello (Fio Cruz Belo Horizonte), and Dr Robert Sebbag (Sanofi Access to Medicines), after an initial global overview of public private partnerships in leishmaniasis from the WHO perspective (Dr D Araw, WHO Geneva) , and a presentation on improving access to treatment for leishmania patients by Dr Sebbag, will focus on some of the clinical and research activity in cutaneous leishmaniasis being performed in Latin America.

Dr Otamires (Fio Cruz Recife) will talk about an integrated management programmme in an area in Pernambuco, which has greatly reduced the impact of the disease. Prof Alexandro Llanos Cuentas (von Humboldt Institute, Lima) will present a retrospective cohort analysis study of CL patients treated with either meglumine antimoniate or sodium stibugluconate).

Prof Bartira Rossi Bergmann (Federal University of Rio de Janeiro) will present early animal results of studies on a topical meglumine ointment being developed by Sanofi.

Finally Dr Byron Arana (TDR/DNDi, Geneva) will present the recently developed criteria for the conduct of leishmania clinical trials in CL, which are designed to accelerate the arrival of new treatments, by ensuring that the data obtained from studies performed, can, if positive, be used to obtain approval from regulatory authorities for the tested compound(s) .

CVs Prof Ana Rabello Prof Ana Rabello is currently Head of the Laboratory of Clinical Research in Infectious Diseases at the Oswaldo Cruz Foundation (Fiocruz), in Belo Horizonte, Brazil. She also coordinates the Leishmaniasis Clinical Reference Center at the Fiocruz unit. She graduated as a Medical Doctor and then specialised in Internal Medicine and Clinical Pathology, completing a Master and a PhD in Tropical Medicine at the Federal University of Minas Gerais state, Brazil. Dr Rabello has been at the Fiocruz, since 1989. She is Professor and Supervisor of the Post graduate Programme of Health Sciences. She also coordinates courses on Ethics of Research involving Humans, and on Leishmaniasis. Dr Rabello has published over 90 articles in peer reviewed journals. Her main research interests are control measures for leishmaniasis, experimental and clinical therapy for leishmaniasis, diagnosis of leishmaniasis and schistosomiasis. Dr Rabello has been the recipient since 1996 of a Scholarship in Research Productivity from the Brazilian National Council Research. She has also been since 2012 the coordinator of the Programme for Innovation for Diagnosis and Drugs for Neglected Diseases of Fiocruz and DNDi, in partnership with the Secretariat of Science, Technology and Strategic Inputs/Brazilian Ministry of Health.

Dr Robert Sebbag Dr Robert Sebbag is Vice President Access to Medicines at Sanofi and leads the development and implementation of the companyâ&#x20AC;&#x2122;s strategy to improve access to medicines in developing and emerging countries. Dr. Sebbag has worked in Brussels for the European Pharmaceutical Industry Association (EFPIA) on creating a communications platform for the pharmaceutical companies operating in Europe. Prior to this, he was Senior Vice President of Communications for the vaccine company, Aventis Pasteur (today known as Sanofi Pasteur). In addition to his activities within the pharmaceutical industry, Dr Sebbag is also teaching Public Health within the Paris hospital system, focusing on tropical parasitic diseases. Dr Sebbag is active within the French Red Cross and has participated in numerous health missions in the Southern Hemisphere. Dr. Sebbag is a Medical Doctor with specialty in tropical parasitic diseases and training in psychiatry.

Dr Daniel Argaw Dr Daniel Argaw is currently Medical Officer for the Leishmaniasis Control Programme in the Department of Neglected Tropical Diseases at the World Health Organization (WHO). Prior to this, Dr Argaw was the National Programme Officer for Disease Prevention and Control at the Ethiopia country office of the WHO. Dr Argaw trained as a Medical Doctor at Addis Ababa University, Ethiopia. He obtained a Master of Public Health from Addis Ababa University and a Master of Advanced Studies in Community Health from the University of Innsbruck, Austria. He has completed numerous international postgraduate trainings including: Communicable

Disease Surveillance (Swiss Tropical Institute, Basel, Switzerland), International Training Course on Tropical Disease Control (Jiangsu Institute of Parasitic Diseases, China), National Health Development and Management of Infectious Diseases (University of Tokyo, Japan) Dr Argaw has extensive experience in disease prevention and control, heath programmes and services management, i.e., planning, implementing, monitoring and evaluation of particularly communicable disease control, and reproductive health programmes at international, national and regional level. Dr Argaw has published over 30 peer reviewed papers or book chapters mainly on leishmaniasis, but also on malaria and HIV/AIDS. Dr Otamires Alves da Silva From 1989 to 1990 Otamires Alves da Silva held a specialization in immunoparasitology at Université de Toulouse III (Paul Sabatier) U.T. III, Toulouse, France and completed her doctorate degree in public health at Université de Paris VI (Pierre et Marie Curie) in 1995. She has published 13 articles in specialized journals, 2 book chapters, 39 works in scientific events, also furnishing information for 15 texts released in newspapers. She has supervised 11 master thesis, 1 PhD thesis, holding a substitute position in 2 others, in addition to having supervised 4 scientific initiation work in collective health care. She acts in the collective health field with focus on public health. In her professional activities she has interacted with 97 coworkers in the coauthorship of scientific papers, participated in the board of examiners in a public examination, in 6 consulting processes, having issued 3 technical opinions and being involved in 7 projects preparation. She is an associate researcher and currently Head of the Leishmaniasis Laboratory at Centro de Pesquisas Aggeu Magalhães/Fundação Oswaldo Cruz.

She is the coordinator of the Leishmaniasis Program in the Department of Parasitology of CPqAM/Fiocruz. In the municipality of São Vicente Ferrer/PE she has coordinated the "Leishmaniasis Program" where she created a "laboratory for endemic diseases research", which was installed and maintained with the financial aid of Sanofi Group France, with whom she has been working since 2002 in the fight against human and animal Leishmaniasis in the State of Pernambuco. She has coordinated the project for the installation of drinking water in 5 municipal schools in St. Vincent Ferrer/PE with the support from "Agence de l'eau Seine Normandie/AESN France. In 2005, a DVD movie related to her research lab and field activities was produced, which is used as an educational material in the health care field. In 2007, she participated as a technical consultant for the launch of a magazine on leishmaniasis in the municipality of Penalva in São Luiz of Maranhão. She wrote a chapter in the book "Les chemins solidaires", released on March 21st, 2008 in Paris, with the presence of the authors. From April 30, 2009, she has been appointed for a five year period as the Leishmaniasis program coordinator in Timbaúba municipality in Pernambuco.

Prof Alejandro Llanos Cuentas Prof Alejandro Llanos Cuentas is currently head of the Leishmaniasis Group, at the "Alexander von Humboldt" Institute of Tropical Medicine of the University Peruana Cayetano Heredia (UPCH) in Lima, Peru. He is also Principal Professor at the Departments of Medicine and Public Health, and Dean of the School of Public Health at UPCH. He was also for 4 years the Director of the Global Fund Initiative in Malaria Control in the Cross Border Areas of the Andean region (Colombia, Ecuador, Peru, Venezuela). Pof Llanos Cuentas trained as a Medical Doctor at UPCH, prior to specializing in Infectious Diseases and Tropical Medicine. He subsequently obtained a Master’s degree in Tropical Medicine at the University of Brasilia, Brazil and a PhD in

Epidemiology at the London School of Tropical Medicine & Hygiene. Prof Llanos Cuentas has over twenty years of experience in the planning, implementation, monitoring and evaluation of research in epidemiology and interventions for the prevention and control of leishmaniasis, malaria and Bartonellosis. He has published extensively in these areas.

Prof Bartira Rossi Bergmann Prof Bartira Rossi Bergmann obtained her BSc in Biomedical Sciences at University of São Paulo (Brazil) in 1981. Then, she got her MSc in Medical Parasitology from LSHTM (UK) in 1982, and PhD in Biology from Brunel University (UK) in 1987 on Immunology of trypanosomatids. In 1994, she carried postdoctoral studies at the University of Toronto (Canada) on tolerogenic vaccination. Presently, she is Professor of Parasitology and Immunology at Federal University of Rio de Janeiro (Brazil) and head of the Immunopharmacology Group. Her interests are focused on antileishmanial chemotherapy and vaccination. Particular attention is paid to the use of nanotechnological tools for sustained drug release from subcutaneous implants, needle free nanosystems applicable to topical and oral chemotherapy, and mucosal vaccination.

Dr Byron Arana Dr Byron Arana is currently Head of the Cutaneous Leishmaniasis Programme at Drugs for Neglected Diseases initiatives (DNDi). He has over twenty years’ experience working on clinical trials and epidemiological studies, mainly in cutaneous leishmaniasis, onchocerciasis, diarrheal and respiratory diseases. Since 2008, Dr Arana has been managing the clinical trials on visceral leishmaniasis that TDR WHO is supporting in the Indian Subcontinent, and participating in the development and implementation of strategies in support of the Indian subcontinent’s visceral leishmaniasis elimination program. From 2000 to 2008, Dr Arana served as Co Director of the Center for Health Studies, at Universidad del Valle de Guatemala, Guatemala and managing the first cooperative agreement between the Universidad del Valle de Guatemala and the Centers for Diseases Control, Atlanta, to support the implementation of multidisciplinary field and laboratory research on tropical diseases in the Central America region. Dr Arana received his medical training at the Universidad San Carlos of Guatemala (1983) and his doctoral degree on Tropical Medicine from the University of Liverpool, UK (1998). Dr Arana has published extensively, especially on leishmaniasis.

SATELLITE SYMPOSIUM 7, 9, 10 WHO/PAHO

[LB01] VISCERAL LEISHMANIASIS OUTBREAK RESPONSE IN SOUTH SUDAN 2009-2012.

ABUBAKAR AM (1), RUIZ-POSTIGO JA (2) , PITA J (1), LUGGA MC (3), BEN-ISMAIL R (2), ARGAW-DAGNE D (4) AND ALVAR J (4): 1:WORLD HEALTH ORGANIZATION, SOUTH SUDAN COUNTRY OFFICE, JUBA, SOUTH SUDAN 2: WORLD HEALTH ORGANIZATION, EASTERN MEDITERRANEAN REGIONAL OFFICE, CAIRO, EGYPT. 3:THE MINISTRY OF HEALTH, REPUBLIC OF SOUTH SUDAN 4: WORLD HEALTH ORGANIZATION, GENEVA, SWITZERLAND.

South Sudan is among the countries with the highest burden of neglected tropical diseases (NTD) worldwide. Visceral leishmaniasis (VL) is a major public health problem in South Sudan, and it is endemic in four states, namely Upper Nile, Unity, Jonglei and Eastern Equatoria. The actual prevalence of VL in South Sudan is unknown due to lack of accurate reporting of the disease. Nonetheless, over 2 million people are considered to be at risk for VL. VL in south Sudan is caused by L. donovani and the implicated vectors are P. orientalis and P.martini. In the past three decades, seasonal VL outbreaks had devastated Sudan and South Sudan, and claimed thousands of lives. The most recent outbreak was confirmed in September 2009 and reached its peak in 2010 and remained protracted until now.

Since September 2009 until December 2012, 28 512 VL new cases and 878 related deaths were reported from 27 treatment centers in 17 counties and four states. Of these cases, 90% (25 939/28 512) were primary VL cases, whereas 9% (2 461/28 512) were secondary cases (relapses and PKDL). Of these new cases, 72% (20 529/28 512) were children less than 17 years of age, 52% (14 826/28 512) were males and 48% (13 685/28 512) females. The overall case fatality rate (CFR) from the 27 treatment facilities was 3.1% (878/28 512). The reported CFR has decreased progressively from the year 2009 until 2012. In 2010, 4.1% (393/9695) was lower than in 2009, 6% (115/1914) (p<0.001). In 2011, 2.5% (293/11 888), was also lower than the previous year (p<0.001). In 2012 the CFR reported was 1.54% (77/5015). This was the first time over the last 20 years that the annual CFR in South Sudan had been significantly reported below 4%.

Timely and well-coordinated multisectoral responses had positively impacted the treatment outcomes and overall reduction of case fatality rate. The response includes the expansion of treatment services, training of health workers on case management, early diagnosis and prompt treatment by using combination of sodium stibogluconate plus paromomycin for 17 days and AmBisome® as second line therapy for relapses, VL/HIV coinfected, severe and other complicated cases. Moreover availability of rapid diagnostic tests, nutritional support services, and health education and social mobilization have also contributed to reduce the impact of this major epidemic.

[LB02] LEISHMANIAISIS CONTROL IN AFRICA IN THE CONTEXT OF THE COUNTRY NEGLECTED TROPICAL DISEASES MASTER PLANS ISSA SANOU1, BONIFACE KINVI1, ADIELE ONYEZE1, LANDRY BIDE1, ALEXANDRE TIENDREBEOGO1, ABATE MULUGETA2, DANIEL A. DAGNE3 1

NTD PROGRAMME, WHO AFRO, BRAZZAVILLE, CONGO,

2

NTD WHO ADDIS ABABA, ETHIOPIA,

3

WHO NTD DEPARTMENT, GENEVA, SWITZERLAND

Leishmaniasis remains a major neglected tropical disease (NTD) in the African Region, which home to about 17% of the global burden of the disease. The visceral form of Leishmaniasis affects mainly East African countries, and the cutaneous form of the disease has been reported in at least 26 countries. Furthermore, in most countries, the disease is co-endemic with other NTDs targeted for control, elimination or eradication in the Region. However, the full scope of Leishmaniasis endemicity in the African Region is not known.

The main challenges related to the control of Leishmaniasis include: (i) establishing and strengthening national Leishmaniasis programme capacity (ii) assessing Leishamaniasis burden in countries, iii) mobilizing adequate resources for Leishmaniasis programme.

In order to accelerate the control of Leishmaniasis and other NTDs, 36 countries have developed multi-year NTD strategic plans, also called “NTD Master Plans” with the support of WHO/AFRO. The Master Plans represent a strong platform for harmonized planning and prioritization and integrated budgeting and resource mobilization. In addition, the Master Plans enable harmonizing interventions, treatments, drug supply, logistics, capacity building, as well as integrated monitoring, surveillance and operations research. The major strategic priorities of the NTD Master Plan include scaling up access to interventions, treatment including Leishmaniasis.

Assessment of the burden of Leishmaniasis in 12 countries by 2014, and the establishment and strengthening of national Leishmaniasis programme in 12 countries by 2015 are the major Regional milestones. The major activities for Leishmaniasis control include: (i) assessment of diseases burden, (ii) developing annual plans, (iii) mobilizing adequate resources for programme interventions, (iv) integration of Leishmaniasis programme with other NTDs, and (v) establishing linkages with HIV, vector control and sanitation programmes.

The availability of NTD Master Plan is an opportunity to enhance Leishmaniasis control in the African Region.

[LB03] CONTROL OF THE LEISHMANISES: GLOBAL PERSPECTIVE DANIEL ARGAW-DAGNE, JEAN JANNIN

INNOVATIVE AND INTENSIFIED DISEASE MANAGEMENT, NEGLECTED TROPICAL DISEASE, WORLD HEALTH ORGANIZATION, GENEVA, SWITZERLAND

The sixtieth World Health Assembly (2007) resolution on leishmaniasis control was a major milestone that urged member states and WHO to reinforce efforts to set up national control programmes. It called for creation of conditions to enable WHO to take a leading role for the initiation, scale up, maintenance and monitoring of national and regional leishmaniasis

prevention and control programmes. The WHO since then has made substantive progresses in framing policy guidelines, setting up regional and national strategic control frameworks, supporting national programmes and building the national and periphepheral health capacities, designing appropriate epidemiological surveillance tools and providing improved global incidence estimate data as well as strongly improving access to drugs. According to the recently published WHO leishmaniasis Global incidence estimate data, although leishmaniasis is prevalent in more than ninety eight countries and three territories, six countries: Bangladesh, Brazil, Ethiopia, India, south Sudan and Sudan harbor 90% of the global visceral leishmaniasis burden. Cutaneous leishmaniasis much more widely distributed in countries in the Americas eastern Mediteranean region, extending from middle east to western and central Asian countries. The series impact of the disease remains underestimated as it is very difficult to get accurate global morbidity and mortality data due to the weak health information and surveillance system in most of the affected countries. The recently adopted NTD road map has clearly stipulated the leishmaniasis control or elimination strategic milestones for the different epidemiological zones of the WHO regions. In this NTD road map it is envisagedto reduce the incidence visceral leishmaniasis to less than one case per 10,000 population per year at sub district level in the endemic countries of South East Asia aiming for elimination as a public health problem by the year 2020. In the other endemic regions high level milestones have been also set by nosogeographical entity to control the three forms of the disease. In order to achieve these milestones and bring impactful changes, a concerted and coordinated effort is required to revamp the control programs, accessing remote endemic areas by scaling up and expanding service delivery and strengthening the surveillance system. Countries should take the lead and ownership of the national programmes to ensure equitable access to services, the continuity of services and achievements of the targets.

[LB04] WHO’S SUPPORT AND COLLABORATIVE EFFORTS TOWARDS THE VISCERAL LEISHMANIASIS PROGRAMME IN ETHIOPIA ABATE M BESHAH1, ABEBAYEHU A MENGISTU1, ISSA SANOU2, DANIEL ARGAW DAGNE3 1:WHO, NTD, Addis Ababa, Ethiopia. 2.:WHO AFRO NTD, Brazzaville 3. WHO, NTD, Geneva, Switzerland.

global estimates of leishmaniasis incidence( 2012). of infection.

[lb05]

OVERVIEW OF CUTANEOUS LEISHMANIASIS IN THE EASTERN MEDITERRANEAN REGION RUIZ POSTIGO JA (1), BEN ISMAIL R (1), ARGAW DAGNE D (2), JANNIN J (2) (1)World Health Organization, Eastern Mediterranean Regional Office, Cairo, Egypt. (2)World Health Organization, headquarters, Geneva, Switzerland

Cutaneous leishmaniasis (CL) is major public health problem in the WHO Eastern Mediterranean Region (EMR). Around 100 000 cases, which represents about 57% of the CL burden worldwide, are reported on average annually. The disease lacks accurate reporting of cases, with actual prevalence estimated at three to five times more than notified. Out of the 23 countries in the EMR, 18 are endemic. L. tropica, mainly transmitted by P. sergenti, causing anthroponotic CL is present in 11 of the endemic countries and L. major, mainly transmitted by P. papatasi, causing zoonotic CL is present in all of them. Few CL cases are also due to other Leishmania species, L. killicki and L. infantum.

Case management for CL in the EMR is not standardized and a well functioning national control programmes have not been set up in most of the endemic countries until recently. Disease control measures have until now been unsuccessful in maintaining a sustained low prevalence of the disease at large scale.

In collaboration with endemic countries WHO has prepared the “Framework for action for cutaneous leishmaniasis in the Eastern Mediterranean Region, 2013 2016”. This document sets five Regional targets on surveillance, case management, prevention, capacity building and research, all of them aiming at reducing the disease burden.

With the aim of standardizing and facilitating case management of cutaneous leishmaniasis in the Region, WHO has prepared a manual focusing on diagnosis and treatment. A step wise algorithm based on size and location of the lesions, causative parasite and immunological status defines three clinical situations for treatment options.

The WHO Eastern Mediterranean Region accounts for the higher burden of cutaneous leishmaniasis worldwide. The disease is complex to control and to treat. The variations in the causing parasite, the transmitting vector and the geographical distribution of the disease across the Region makes it difficult for a single, ready to use method of control and treatment. The

new framework for action and the manual for standardized case management of cutaneous leishmaniasis need to be adapted based on the endemic countries eco epidemiological profile and prevailing health system context.

[LB-06]

LEISHMANIASIS IN THE WHO EUROPEAN REGION: EPIDEMIOLOGICAL PROFILE

L. infantum, P.alexandi, Ph.kandelakii,Ph. balcanicus,Ph.sergenti, Ph.smirnovi,Ph.mongolensis,Ph. chinensis, Ph.syriacus,Ph. papatasi, Ph. halepensis, Ph. syriacus, Ph. longiductus, Ph. perfiliewi, Ph. perniciosus, Ph. ariasi, Ph.tobbi, Ph. arabicus, Ph. neglectus.

L. tropica P. sergenti

L. major L.

infantum

SATELLITE SYMPOSIUM 11 Hertape Calier

Title: Biology of A2 molecules in Leishmania Author name(s): Laura Isobel McCall, Wen Wei Zhang, Greg Matlashewski Author affiliation(s): McGill University, Department of Microbiology and Immunology, Montreal, Canada Abstract text

Searching for genes that are expressed at high levels in amastigotes and at low levels in promastigotes can highlight factors that are important for mammalian infection. The A2 gene family was the prototypical such amastigote specific gene to be indentified in Leishmania donovani. A2 expression is induced during promastigote to amastigote differentiation, and conversely A2 levels are low under normal promastigote culture conditions. Downregulation of A2 in L. donovani, either by antisense mRNA transfection or by partial knockout of some A2 genes, significantly decreased parasite capacity to survive in the visceral organs, thereby confirming the role of A2 in visceral infection. In contrast, A2 is a pseudogene in Old World cutaneous species (L. major and L. tropica). Restoring A2 expression in L. major enhanced its ability to survive in the visceral organs while decreasing cutaneous infectivity. These results indicate that A2 is a major determinant of visceral disease and is sufficient to enhance cutaneous species survival in the visceral organs. Recent work highlights mechanisms by which A2 may promote visceral organ survival: in addition to being induced by promastigote to amastigote differentiation, A2 is strongly induced by a number of stresses, including heat shock, misfolded protein stress and induction of the unfolded protein response. A2 also protects against heat shock and oxidative stress. Overall, these results suggest that visceral organ infection is more stressful for the parasite than cutaneous infection; A2 expression allows visceral species to survive the stresses encountered in the visceral organs. A2 is therefore a major determinant of the ability of Leishmania to infect the visceral organs.

Title: The antigen "A2" and the production of recombinant vaccines Author name(s): Ana Paula Fernandes Author affiliation(s): Faculdade de Farmรกcia, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil Abstract text Visceralizing factors of Leishmania may also constitute potential species specific vaccine candidates.

Having this in mind and aiming to develop a vaccine against visceral leishmaniasis targeting the intracellular amastigotes, which is the parasite stage that persists throughout infections, we have

tested the amastigote specific antigen (A2) as a vaccine candidate. The A2 antigen has been extensively characterized, its coding sequence has been optimized and different vaccine formulations containing A2 different adjuvants or A2 expressed by attenuated adenovirus and non virulent trypanosomatids have been developed, which were shown to be protective in mice, dogs and non human primates. After transfer of technology between the Federal University of Minas Gerais and the company Hertape Calier, a recombinant protein formulation containing saponin as adjuvant has been produced, approved by regulatory public authorities and tested in clinical trials in dogs. The technical and scientific steps and aspects followed for development of these recombinant A2 vaccines will be further presented and discussed in the satellite symposium.

Finnancial Supporte: INCTV, FAPEMIG, CNPq and Hertape Calier

Title: Clinical Trial Phase III Study of the efficacy of the vaccine Leish Tec© Author name: George Luiz Lins Machado Coelho Author affiliation: Escola de Medicina, Universidade Federal de Ouro Preto.

Abstract text Brazil is an endemic country for zoonotic visceral leishmaniasis (ZVL) and regularly conducts prophylactic and epidemiologic control programs involving treatment of human cases, insect vector control and the removal of seropositive dogs. Dogs have a fundamental role in urbanization and in the maintenance of the disease in endemic areas. In the urban and rural areas of municipal of Porteirinha, a city near of Montes Claros, north of Minas Gerais state of Brazil, were accomplished a field trial set out in randomized blocks to evaluate the efficacy of Leish Tec® vaccine against canine visceral leishmaniasis (CVL). Blood samples were collected of 2,629 dogs distributed in all 14 urban areas (2,015 dogs) and six of 11 rural districts (614 dogs). The prevalence of CVL were 20.6% (9.1 55.3) and 73.3% (35.9 92.8) in the urban and rural areas, respectively. After serologic survey, a sample of 865 negative dogs by ELISA (Biomanguinhos), and IFAT (Biomanguinhos) were selected to receive three doses and one booster of Leish Tec® vaccine or placebo. In the highest infection areas, 150 negative pre vaccinated (or placebo) beagle dogs were domiciled as sentinels’ dogs. These 1,015 dogs were follow up during 527 days after the third dose. Dogs seropositive by ELISA, IFAT and Kalazar Detect rapid test (KD rK39) (IMBIOS) were removed from the area for necropsy and parasites diagnosis (direct exam, marrow culture, PCR of tissue and xenodiagnosis). The protective effect for the transmission of Leishmania in sandfly was 40% in animals vaccinated with Leish Tec ® that produced antibodies A2. Considering only the parasitological diagnosis results, the protective effect of the vaccine Leish Tec® in this population was 71% (34.9 87.3; p value=0.001), and 81% (CI: 37.6 94.1, p value=0.001) when the analysis was stratified by individual response of the dog to the antigen A2, i.e., the efficiency of the vaccine is increased in those animals seropositive for anti A2. These results demonstrate that the vaccine Leish Tec® could potentially be employed for vaccination in conjunction with other health strategies, such as serologic detection and euthanasia of infected dogs. Supported by Hertape Calier Saúde Animal S.A.

Title: Strategies for Vaccine Leish Tec use as a tool for prevention and control of LVC in Brazil.

Author name: Dr. Francisco Anilton Alves AraĂşjo Author affiliation: Hertape Calier SaĂşde Animal â&#x20AC;&#x201C; Brazil Abstract text

There are many changes that need to be implemented to improve the surveillance and control of visceral leishmaniasis in Brazil. Deploying Test DPP, recommending the use of collar impregnated with insecticide and vaccination of pet dogs with vaccine Leish Tec, aims at the detection of positive animals with the removal of the same, protection of susceptible and comes in intuit replacing the strategy of euthanasia animal. Brazil is the only country in the world to recommend euthanasia of dogs positive for leishmaniasis. The vaccine Leish Tec has a protection level of 96.4% in vaccinated animals effectively and has been considered as an important tool to prevent and control the disease. This strategy should be associated with the other already recommended by the Ministry of Health control vaccinated animals should be done by microchipagem to avoid mistakes and livestock trade. Vaccination in areas of intense transmission should be seen as a priority. Areas where it has been detected the first cases of canine visceral Leishmaniasis has also shown interest in performing vaccination in order to prevent the disease from spreading throughout the municipality.

SATELLITE SYMPOSIUM 13 Zoetis/Pfizer

Simpósio Zoetis: Atualizações e novas pesquisas a cerca da Leishmaniose Visceral Canina.

May 16, 2013 Satellite Symposium 2 – 4 p.m.

1) Imunopatogenia e Sinais Clínicos da Leishmaniose Visceral Canina (25 minutos) Palestrante: Paulo Tabanez

Médico Veterinário graduado pela Universidade Estadual de Londrina (UEL/PR 2001). Pós graduado em Clínica e Cirurgia de Pequenos Animais pelo Centro Regional Universitário de Espírito Santo do Pinhal/SP (2004). Mestre em Imunologia pela Faculdade de Medicina da Universidade de Brasília/DF (UnB 2006). Diretor do Hospital Veterinário Prontovet (DF), Coordenador dos departamentos de Oncologia e Infectologia. Experiência na área de Medicina Veterinária, com ênfase em Imunologia, Infectologia e Oncologia, atuando principalmente nos seguintes temas: Imunopatolgia, Imunomoduladores e Leishmanione Visceral Canina. Membro fundador do BRASILEISH. 2) Indicadores Clínicos e Laboratoriais da Infectividade Canina (20 minutos) Palestrante: Vitor Márcio Ribeiro

Médico Veterinário graduado pela Universidade Federal de Minas Gerais (UFMG/MG 1980).

Mestrado em Medicina Veterinária pela Universidade Federal de Minas Gerais (UFMG/MG 1988) e doutorado em Parasitologia pela Universidade Federal de Minas Gerais (UFMG/MG 2001). Professor adjunto IV da Pontifícia Universidade Católica de Minas Gerais (PUC/MG), nas disciplinas de Clinica Médica de Cães e Gatos, Doenças Infectocontagiosas de cães e gatos e Extensão Rural. Diretor técnico na VISIOVET Diagnóstico Veterinário e Diretor Clínico da Clínica Veterinária Santo Agostinho. Experiência na área de Medicina Veterinária, com ênfase em Doenças Parasitárias de Animais, Clínica e Cirurgia Geral, Neurologia Clínica e Cirúrgica atuando principalmente nos seguintes temas: Leishmaniose Visceral Canina, Clínica e Cirurgia Geral e Neurológica de Cães e Gatos. Presidente do BRASILEISH. 3) Diminuição da carga parasitária em Medula Óssea em cães vacinados com Leishmune®. (20 minutos) Palestrante: Adriane Pimenta da Costa Val Bicalho

Medica Veterinária graduada pela Universidade Federal de Minas Gerais (UFMG/MG 1990).

Mestrado em Medicina Veterinária pela Universidade Federal de Minas Gerais (UFMG/MG 1994) e doutorado em Ciência Animal pela Universidade Federal de Minas Gerais (UFMG/MG 2004). Professora adjunto na Escola de Veterinária da Universidade Federal de Minas Gerais (UFMG/MG) nas disciplinas Semiologia e Clínica Médica de Pequenos Animais, com ênfase em Dermatologia. Áreas de pesquisa: dermatologia de pequenos animais, leishmanioses dos animais domésticos e bem estar animal.

4) Alterações imunológicas em leucócitos do sangue periférico de cães imunizados com vacinas de primeira e segunda geração contra a Leishmaniose Visceral Canina (25 minutos) Palestrante: Olindo Assis Martins Filho

Graduado em Farmácia Bioquímica pela Universidade Federal de Ouro Preto (UFOP/MG 1989).

Mestrado em Bioquímica e Imunologia pela Universidade Federal de Minas Gerais (UFMG/MG 1992), Pesquisador Visitante na Johns Hopikns University (1994), doutorado em Bioquímica e Imunologia pela Universidade Federal de Minas Gerais (UFMG/MG 1997) e pós doutorado

pela The University Of Texas Houston, UTH, Estados Unidos (USA 1999). Pesquisador Titular da Fundação Oswaldo Cruz e Chefe do Laboratório de Biomarcadores de Diagnóstico e Monitoração do Centro de Pesquisas René Rachou/FIOCRUZ Minas Gerais/MG. Experiência na área de Imunologia Celular e Molecular, com ênfase na busca de novas metodologias aplicadas ao diagnóstico, imunidade vacinal e monitoração de doenças infecto parasitárias.

INDEX AUTHOR AARA, N. ABANADES, D.R. ABASSI, I. ABATE, H.L. ABBEHUSEN, M.M. ABBEHUSEN, M.M.C. ABBOUD, L.C.S. ABDALLAH, A. ABDALLAH, A.A. ABDEEN, Z. ABDEL HALEEM, A.A. ABDELADHIM, M. ABDESSALEM, C.B. ABDOLI, H. ABDOUN, A.O. ABEBE, T. ABENGOZAR, M.A. ABI ABDALLAH, D.S. ABID, H. ABRAHAM, T. ABRANTES, C.F. ABREU SILVA, A.L. ABREU, B.L. ABREU, E. ABREU, H.C.N. ABRIL, M. ACARDI, S.A. ACOSTA, L.A. ACUÑA, V.L. ADAMS, E. ADAMS, E.R. ADAUI, V. ADDYA, S. AEBISCHER, T. AFONSO, L.C.C. AFONSO, M.M.S. AFONSO, M.O. AFRIN, F. AGRA, M.C.R. AGRA, M.F. AGUADO, J.M. AGUDELO, D.

ABSTRACT O 171 P 013 P 093 O 216 P 713, P 715 P 223 P 038 P 392 P 691 P 692 O 243, O 290, P 455, P 684 P 288 O 160, P 039, P 958 P 958 O 318 P 610 P 410 O 133 P 461, P 464 P 475 O 159 P 517, P 580 O 003 P 145 P 507 P 812 O 151 P 233, P 554 P 825 O 045, O 097 O 011, O 380, P 904 P 704 O 060, O 258, P 332, P 361, P 458, P 794 P 151 P 586 O 139, P 144, P 382, P 397, P 438 P 333 O 162 P 138 P 725 O 219 P 466 P 782 P 440 O 016 O 380, P 904

AGUIAR SOARES, R.D.O. AGUIAR SOARES, R.D. AGUIAR SOARES, R.D.O. AGUIAR, B.G. AGUIAR, C.O.M. AGUIAR, M.G. AGUIAR, S.B. AGUIAR, V.B. AGUILAR, E. AGUILLERA, A.H. AHMADI, F. AHMED, B. AHMED, E.M. AINSLIE, K.M. AISSI, W. AJOUAD, M. AKHAVAN, A.A. AKOPAYANTS, N. AKOPYANTS, N.S. AL RIMAWI, F. AL DWIBE, H. AL HAZZA, I.M. AL JAWABREH, A. AL JAWABREH, H. AL KHALIFA, M.S. ALAGESAN, K. ALAKURTTI, S. ALAM, M.Z. ALAM, S. ALAM, S.M. ALARCON, G. ALBA, M. ALBAN, S. ALBANEZ, M. ALBANO AMORA, S.S. ALBERGANTE, L. ALBERICIO, F. ALBERIO, S.H.V.O. ALBEROLA, J. ALBERTINI, A. ALBUQUERQUE, A. ALBUQUERQUE, E.R. ALCAZAR, W. ALCOLEA ALCOLEA, P.J. ALCOLEA, P.J. ALCOVER, M.M. ALENCAR, A.A.

P 072, P 143 P 071, P 533 P 129 P 197, P 199 P 276 O 233, O 234 P 587, P 588 P 076 P 860 P 046, P 342 O 305 O 064 O 084, O 222, O 376 O 023 O 308 P 355 O 253 O 195, O 127, O 318 O 348 O 074, O 142, O 351 P 455 P 724 P 014 O 243, P 684 O 243, P 684 P 014 P 019 P 338 P 026, P 724 O 055 O 222 P 704 P 397, P 438 P 826 O 311 P 865, P 868, P 872, P 875 P 874, P 878, P 879 O 340 P 467 P 935, P 943 P 075, P 646 P 366 P 463, P 503 P 939 P 338 P 157 O 348 P 148, P 152 P 944, P 947 O 170, P 583

ALENCAR, C.H.M. ALEXANDER, N. ALEXANDRA, W.A. ALEXANDRE MOREIRA, M.S. ALEXANDRE MOREIRA, M.S. ALEXANDRE PIRES, G. ALEXANDRE, E.C. ALEXANDRE, J.L.A. ALEXANDRE, T.R. ALFANI, A.O.S. ALFONSO, E. ALFONZO, E. ALI, N. ALI, V. ALIANCA, A.S.S. ALIGIANNIS, N. ALIMOHAMMADIAN, M.H. ALIROL, E. ALKAN, M.Z. ALLA, L. ALLEGRETTI, S.M. ALLEN, R. ALLENSPACH, K. ALMEIDA ALVES, T.M. ALMEIDA NETO, W.S. ALMEIDA AMARAL, E.E. ALMEIDA SOUZA, F. ALMEIDA, A.F. ALMEIDA, A.J. ALMEIDA, A.P. ALMEIDA, A.S. ALMEIDA, B.F. ALMEIDA, C. ALMEIDA, E.L. ALMEIDA, G.G. ALMEIDA, J.A. ALMEIDA, J.F. ALMEIDA, L. ALMEIDA, L.F. ALMEIDA, M.A.C. ALMEIDA, M.C.M. ALMEIDA, M.S. ALMEIDA, N.A. ALMEIDA, N.J. ALMEIDA, R. ALMEIDA, R.A.

O 043, P 739, P 853 O 008 P 848 P 440, P 594 P 593 P 488, P 528, P 571 O 182, O 341 P 398 P 293, P 296, P 608, P 609, P 614, P 617 P 351, P 389 P 095 P 257 O 375 O 154 P 599, P 603 P 638, P 644 P 487 O 318 O 356 O 295 P 843 P 911, P 912 O 192 P 646 P 632 O 322 P 346, P 408 P 507 P 411 O 312 P 296, P 608, P 614 P 682 P 720 P 936, P 948, P 954 P 771 P 054 P 786 P 219, P 293, P 296, P 608, P 609, P 614, P 617 O 294, P 393 P 740 O 157, P 123 P 445 P 133 P 076 P 821 P 192 P 550 P 570 O 036, P 074, P 326 O 209, O 210, O 328 P 207

ALMEIDA, R.P. ALMEIDA, T.C. ALMEIDA, T.M. ALMEIDA, V.A. ALONSO, A. ALONSO, C. ALONSO, D.P. ALSAADI, M. ALVAR, J. ALVAREZ, E. ALVES FERREIRA, E.V.C. ALVES, A. ALVES, B.G.V. ALVES, C. ALVES, C.R. ALVES, E.B. ALVES, F. ALVES, F.P. ALVES, K.F. ALVES, M.A. ALVES, M.L. ALVES, M.L.R. ALVES, N.D. ALVES, N.M.E. ALVES, P.B. ALVES, P.H. ALVES, P.M. ALVES, R.J. ALVES, S.S. ALVES, T.M.A.

ALVES, W.A. ALZATE, J.F. AMARAL, A.C.F. AMARAL, D.N.D. AMARAL, E.E.A. AMAROUCH, H. AMATO, V.S. AMIRPOUR, S. AMIT, A. AMOAH, S. AMOR HAOUAS, N. AMORA, S.S.A. AMORIM, A.A.S.

P 323 P 250, P 456, P 481, P 740 P 073 P 421 P 265, P 411 P 639, P 719 P 148, P 152, P 157 P 113 O 354, P 057, P 173 P 450 O 106, O 202, O 016, O 230, O 380 O 267 P 198, P 312 P 476 P 516 P 629 P 629 P 065, P 248, P 550 P 689 O 365 P 588 P 538, P 553 P 488 P 915 P 292 P 917 P 194 P 069 P 865, P 868, P 872, P 874, P 875, P 878, P 879 P 609 P 481 P 504 P 747 P 297, P 623 P 643 P 698 P 516 P 885, P 889, P 891, P 894 P 871, P 886, P 888, P 890, P 897 O 283, O 307 P 507 P 445 P 571 P 545 P 016 P 269 O 168, P 435 P 701 P 063 P 487 P 908 P 678 P 439

AMORIM, C.F. AMORIM, I. AMORIM, I.F. AMPUERO, J.S. AMRI, F. AMRO, A. ANDERSON, B. ANDRADE FILHO, J.D. ANDRADE FILHO, J.D. ANDRADE NARVAEZ, F.J. ANDRADE NETO, V.V. ANDRADE, A.S.R. ANDRADE, B.B. ANDRADE, D. ANDRADE, D.R. ANDRADE, H.F.

ANDRADE, H.M. ANDRADE, J.D. ANDRADE, J.M. Andrade, J.M. ANDRADE, L.C. ANDRADE, L.D. ANDRADE, M.B. ANDRADE, M.C. ANDRADE, M.M.C. ANDRADE, M.S. ANDRADE, M.V. ANDRADE, P.H. ANDRADE, P.H.R. ANDRADE, P.P. ANDRADE, P.S. ANDRE, M.R. ANDRES, C. ANDREU, D. ANDREWS, N. ANDREWS, P. ANDRIC, B. ANDRIGUETTI, M.T.M. ANEES, M. AÑEZ, N. ANG, K.H. ANGEL, S.O. ANGULO, C. ANIBA, M.R. ANNAJAR, B.

P 865, P 868, P 872, P 875 P 678, P 874, P 878, P 879 P 309, P 310, P 317 P 606, P 839 P 134 P 355 P 092, P 455, P 684, P 724 O 348 P 798, P 812, P 846, P 893 P 818 P 647 P 120 P 552 O 294, P 416 P 446 O 051, P 136, P 187 P 022 P 622 P 206, P 213, P 537, P 562 P 066, P 200, P 214, P 349, P 795 P 057 P 442 P 339 P 099 P 149, P 498 P 118 P 496 P 398 P 293, P 614, P 789 P 824 P 520 P 362, P 376 P 842 P 176 P 190, P 249 P 628 P 877 P 475 O 348 O 340 O 013, P 426 P 748 O 124 P 836 O 285 P 341 P 438 O 345 P 724

ANNELIES, M. ANNUNZIATELLA, M. ANTONELLI, L.R. ANTONELLI, L.R.D.V. ANTONIALLI, S.A.C. ANTONIOU, M. ANVERSA, L. AOKI, J.I. AOUN, K. AQUINO, M.C.C. AQUINO, T.A. ARAGAO, M. ARAGAO, M.T. ARAMIN, S. ARANA, B.A. ARANA, F.E. ARANDIAN, M.H. ARANGO DUQUE, G. ARAUJO NETO, A.R. ARAUJO SANTOS, T. ARAUJO, A. ARAUJO, A.I.F. ARAUJO, A.M. ARAUJO, C.F. ARAUJO, F.A.P. ARAUJO, F.M. ARAUJO, I.B. ARAUJO, J.M. ARAUJO, J.M.P. ARAUJO, K.C. ARAUJO, L.H. ARAUJO, M.F.L. ARAUJO, M.I.A. ARAUJO, M.S.S. ARAUJO, M.V. ARAUJO, N.N.S.L. ARAUJO, P.S.R. ARAUJO, R.N. ARAUJO, R.S. ARAUJO, V.E.M. ARAUJO, V.P. ARCE, A. ARCE, L. AREVALO, J. ARGAW, D. ARGYROPOULOU, K. ARIELLA DERENGE, C.

O 146, P 451 P 048 P 165 P 253 P 808 O 135 P 462 P 637 P 355, P 484, P 656, P 790 O 119 P 402 P 428 P 546 O 151 P 740 O 216 P 605 P 605 O 195, O 127, O 318 O 034 P 572, P 619 P 322 O 051, P 013 P 840 P 293, P 296, P 608, P 614, P 617 P 054, P 056 P 282 P 854 P 935 P 939 P 719 P 880 P 815 P 176 P 211 P 395 P 631 P 601 P 141 P 516 P 194 P 292 P 488, P 528, P 571, P 593, P 594 P 921 P 149 P 239 O 317 P 935 P 816, P 945 P 900 P 521 P 769 P 008 O 139, P 144, P 397, P 438 O 351, O 254, O 267 O 202 P 487 O 320

ARISTIDES, S.M.A. ARJMAND, R. ARKIN, M. ARKIN, M.R. ARMAND, A. ARMELIM, I.M. ARONSON, N. ARORA, S.K. ARROYO, F.A. ARRUDA, C.C. ARRUDA, C.P. ARRUDA, E.S. ARRUDA, L.V. ARRUDA, M.M. ARRUDA, P. ARRUDA, S. ARSERIM, S.K. ASEFFA, A. ASELA, C. ASELA, C.X. ASHOURI, H. ASLAN, H. ASRAT, H. ASSIS LIMA COSTA, F. ASSIS, A.P. ASSIS, E.W.S.D.V. ASSIS, M.D.G.G. ASSIS, T.S.M. ASSUNCAO, R.S. ATA, A. ATASOY, A. ATELLA, G.C. ATHAUDA, I. ATTIA, H. ATTIPOE, I. AUGUSTO, J. AULNER, N. AURELIANO, D.P. AVELAR, D. AVELAR, D.M. AVERSI FERREIRA, T.A. AVILA, G. AYALA URDAPILLETA, A.A. AYALA, M. AYALA, M.S. AYAZ, S. AYEH KUMI, P. AZADMANESH, K.

P 554 P 301, P 913 O 306 O 285 P 624 P 955 O 160 P 114 P 918 P 202 P 521 P 093 P 093 O 296, P 929 P 884 P 480, P 527 O 295 P 934 O 364 O 365 P 621 O 160, O 161, O 320, O 026 P 627 P 091, P 341 P 759 P 803, P 807 P 760, P 762 P 358 P 664 P 272 O 321 P 166 P 670 P 058, P 059, P 922 P 946 O 372 O 284 P 431 P 145 P 700, P 703, P 764 P 828 O 380 P 105 P 709 P 536 O 124 P 946 O 239

AZEREDO COUTINHO, R.B.G. AZEVEDO JUNIOR, J.W.C. AZEVEDO, C.G. AZEVEDO, E.G. AZEVEDO, F.S. AZEVEDO, L. AZMI, K. B, V.M. B. DE JESUS, J. B. LUCHEIS, S. BABBA, H. BABIKER, D.E. BACCI, L. BACELLAR, M.O. BACELLAR, O. BACELLAR, O.A. BACHELDAR, E.M. BACHMAIER, S. BAEZ, P. BAEZA RAMIREZ, I. BAFICA, A. BAFICA, A.M. BAGAR, S.R.A. BAHARIA, R. BAHARIA, R.K. BAHIA, A.P.C.O. BAHIA, D. BAHIA, E.M. BAHIA, M. BAHIA, M.T. BAHRIANI, M. BAIAO, A. BAILEY, M. BAILONA, G.H.C. BAILOR, H.R. BAKHIET, S.M. BALASEGARAM, M. BALASEGRAM, M. BALDEVIANO, G.C. BALEELA, R. BALLART, C. BALNO, C. BALTAZAR, F.H. BALTUSSEN, R. BANDARA, S. BANDARRA, M.B. BANDEIRA, C. BANDEIRA, D.S.

O 274, O 165 P 468 P 725 P 606, P 839 O 082 P 845, P 847 P 179, P 804, P 811 P 392 O 290, P 455, P 671, P 684 O 238 P 243 P 863 P 140, P 905, P 908 P 288 P 481 P 021 O 272 O 278, P 023 P 294 O 308 P 101 P 825 P 595 O 291 P 141 P 353 P 043 O 089, P 041 P 381 P 393 O 030 P 630, P 931, P 932 O 180 P 795 P 591 P 179, P 804, P 811, P 847 P 061 P 856 O 092, O 027 P 288 P 627, P 691, P 692 O 356 O 351 P 835 O 102 P 944, P 947 O 131 P 054 P 691, P 692 P 670 P 177, P 190 P 643 P 681

BANDEIRA, E.L. BANDEIRA, M.C.A. BANDINI, G. BANDOPADHYAY, S.K. BANIC, D.M. BANIHASHEMI, M. BANKOTI, R. BANSAL, M. BANULS, A.L. BAÑULS, A.L. BAQUERO, O.S. BAQUI, M.M.A. BARALDI, S.R. BARATA, I.R. BARBAS, C. BARBIAN, K.D. BARBIERI, C.L. BARBO, M.L.P. BARBOSA DOS, M.L. BARBOSA JR, W.L. BARBOSA, A.D. BARBOSA, D.S. BARBOSA, F.M.C. BARBOSA, G.L. BARBOSA, I.R. BARBOSA, J.A.R. BARBOSA, J.C. BARBOSA, J.C.O. BARBOSA, J.E.R. BARBOSA, J.R. BARBOSA, M.G.V. BARBOSA, P.R.A. BARBOSA, S.R. BARBOSA, V. BARBOSA, W.O. BARCELLOS, N.M.S. BARCINSKI, M.A. BARHOUMI, M. BARHOUMI, W. BARKER, J.H. BARON, S. BARRA, A.

BARRAL NETTO, M. BARRAL, A.

P 006 P 673, P 734 O 075 P 802 P 210 P 340 O 371 P 449 O 366 P 140 P 305, P 306 P 937, P 938 P 216 P 489 P 864 P 373, P 461 P 029 P 175, P 350 O 030 P 955 P 261 P 498 P 552 P 708, P 723 P 156 O 377 P 830 P 767 P 924 P 925 P 926, P 927 P 767 P 489 P 929 P 583 P 956 P 865, P 868, P 872, P 874, P 875, P 878, P 879 P 309 P 842 P 509 P 033 P 044 P 403, P 517 P 654 O 302 P 944 P 024 O 238, O 291, P 038, P 093, P 118, P 132, P 187, P 721, P 844 O 238, O 326, O 250, O 291, P 038, P 118, P 132, P 166, P 187, P 198, P 223, P 283, P 343, P 618, P 844

BARRAL, M.A.D.P. BARREIRO, E.J. BARREIRO, E.J.L. BARREIROS, D.C. BARRETO, A.S. BARRETO, T. BARRETO, T.V. BARRIO, A.B. BARRIOS, M.A. BARROS, A.S. BARROS, J.A.C. BARROS, N.B. BARROS, T.L. BARROSO, P.A. BART, A. BART, O. BARTHOLOMEU, D.C. BARUD, H.S. BASAGLIA, F.C. BASOMBRIO, M.A. BASSO, M.A. BASTIEN, P. BASTOS, C.E. BASTOS, L.A. BASTOS, V.J.C. BATE, W. BATES, M.D. BATES, P. BATES, P.A. BATHIA, A. BATISTA, A.J.S. BATISTA, A.P. BATISTA, J.A.S. BATISTA, J.F. BATISTA, L.R. BATISTA, M.A. BAUGH, L. BAVIA, M.E. BAYIH, A.G. BEATTIE, L. BECKER, K.M. BECKMANN, A.M. BEGO, A. BEHAR, P.R.P. BEIJNEN, J.H. BEITING, D.P. BEJARANO, E.E. BELAUNZARAN, M.L.

P 721 P 571 P 488 P 833 P 576 P 310 P 245 O 038 P 914 P 892 P 760 P 108 P 780 O 038 P 485 O 255, P 061 O 163 O 257, P 035, P 181, P 244, P 486, P 491 P 433 P 231 P 485 O 038 P 772 O 014, O 221, O 351 P 883 P 391 P 675 P 055, P 697 P 946 O 332, O 042, O 318 O 158, O 194 P 946 P 576 P 555 P 795 P 489 O 198 P 462, P 781, P 803, P 807 P 509 P 138 O 107, O 348 O 039 O 100, P 833 O 117 O 340 P 523 O 093 P 295 P 870 O 078, O 111 O 241, O 087 P 951 P 334

BELAY, S. BELHAJ HMIDA, N. BELIZARIO, D. BELL, A.S. BELLO, A. BELLO, A.R. BELLO, F.J.G. BELO, V.S. BELOTI, C.C. BEN ABDA, I. BEN ABDERRAZAK, S.B. BEN AHMED, M. BEN KHALAF, N. BEN MESSAOUD, N. BEN SGHAIER, I. BEN ABID, M. BEN OTHMAN, R. BEN SGHAIER, I. BENASSI, J.C. BENCHIMOL, M. BENETI, S.A.A. BENHELEL, K. BENIE, G. BENIGNO, C.V. BENITEZ, A.D.N. BENKAHLA, A. BENSAID, M. BENTES, P. BENTO, E.C. BERCINI, M.A. BERG, M. BERGMANN, B.R. BERL, F.P. BERMUDEZ, H. BERNARD, H.I. BERNARDINETTI, U. BERNARDINO, A.M.R. BERRICH, M. BERRIMAN, M. Berriman, M. BERROSPE, P.E. BERROZPE, P. BERROZPE, P.E. BERTHO, A.L. BERTOLLO, C.M. BERTOLLO, D.M.B. BESCHIN, A. BETTAIEB, J.

O 216 P 942 P 153 P 909, P 910 O 152 O 342 P 217 P 065 P 723 P 683, P 708 P 540 P 355 P 586 P 153, P 958 O 306 O 363 P 355 P 484 O 345 P 484 P 539 P 103 P 631 P 355 O 125 P 695 P 713, P 715 P 059 P 501 P 189 P 781 P 870 O 143, P 382, P 384, P 385, P 386 P 162, P 304, P 407, P 545, P 547, P 559, P 574, P 575 P 371 P 380 P 008 P 957 P 427, P 523 O 124 O 002, O 143, P 672 O 298 O 344 P 778 P 814 P 806 P 228 P 238 P 150, P 237 P 632 P 857 P 101 P 942 P 916 P 305, P 306, P 922

BETTAÏEB, J. BETTS, M.R. BEVERLEY, S. BEVERLEY, S.M. BEZERRA DE MENEZES, J.G.P. BEZERRA, A.C.D.S. BEZERRA, C.B. BEZERRA, F.M.B.V. BEZERRA, G.F.B. BEZERRA, I.O. BEZERRA, I.P.S. BEZERRA, M.E. BEZUNEH, A. BHANDARI, V. BHANDERI, P.K. BHASKAR, B. BHASKAR, K.R.H. BHATIA, A. BHATTACHARJEE, H. BHATTACHARYA, P. BHATTACHARYA, S.M. BHATTACHARYYA, T. BHATTARAI, N. BHATTARAI, N.R. BHOWMICK, S. BHUIYAN, A.T. BIANCO, E.M. BICHAUD, L. BIFELD, E. BIGELI, J.G. BILCH, K. BIMAL, S. BINO, S. BIONDO, A.W. BISETTO JR., A.B. BISETTO JUNIOR, A. BISOL, J. BISWARO, L. BITTAR, R.C. BITTENCOURT, C.R. BIVONA, M. BLACKBURN, G.J. BLACKWELL, J. BLACKWELL, J.M. BLAKE, D. BLANCO, V.M. BLANDON, G.A.

O 125 O 087 O 297, O 348 O 074, O 075, O 142, O 351, P 256, P 277 O 129 O 052, P 112, P 115, P 116 P 212 P 678 P 303 P 565 O 112 P 436, P 729, P 730, P 732, P 733 P 956 O 116, P 208 P 897 O 216, P 610 P 363, P 364 P 753 P 003 O 266 O 093, P 610 O 004 O 026 P 388 O 265 O 143, O 078, P 053, P 672 O 111, O 067, P 383, P 386, P 679 O 154 O 154 O 109 P 638, P 644 O 194 O 105 P 676 P 410 P 063 P 599 P 295, P 611, P 843 P 395, P 852 P 776 O 096 P 826 P 854 P 500 P 269 P 561 O 187 P 053 P 133, P 264 O 343 O 352 O 268, P 011 P 646 O 147 O 099, P 737 P 589

BLANK, A.F. BLAYNEY, B. BLONDEL, A. BLOOM, J. BLUM, J. BOAKYE, D. BOAMPONG, J. BOAVENTURA, V. BOCCHI, M.R.

BOELAERT, M. BOELART, M. BOGAART, E.V.D. BOGDAN, C. BOGDAN, C.M. BOGGILD, A. BOGGILD, A.K. BOIREAU, P. BOITE, M.C. BOIX, E. BOLAS, A.S. BOLETI, H. BOLHASSANI, A. BONANO, V. BONFIETTI, L.X. BONFIM, T.D. BONGIORNO, G. BONGRAND, P. BORCHANI, L. BORCHICHI, S.R. BORDA, C.E. BORDIGNON, M.O. BORDONI, M. BORGES, A. BORGES, A.C.J. BORGES, D.T. BORGES, R. BORGES, V. BORGES, V.M. BORJA CABRERA, G.P. BORJA, L.S. BORST, P. BOSHART, M. BOSQUIROLI, L.S. BOSQUIROLI, L.S.S.

P 481 P 600 P 403 O 085 P 061 P 946 P 946 O 238, P 343 P 857 O 012, O 078, O 111, O 015, O 313, O 316, O 357, O 055, O 163, O 067, O 262, O 263, O 265, O 267, O 268, O 269, P 144, P 361, P 679, P 691, P 692, P 749, P 753 O 315 P 332 O 064 O 132 O 254 P 397 O 221, O 124 P 073 P 866 O 136, P 119 P 475 P 185 P 318 O 239 O 282 P 540 P 294 O 212 P 207 P 184, P 517, P 580 P 008 P 357, P 666, P 779 P 801 O 044, P 391, P 526 P 638, P 644 P 051 P 950, P 956 O 259, P 542, P 797, P 909 P 136 P 106 P 036, P 118, P 343 P 548 P 433 O 275, O 326, O 051, P 013 P 918 O 044, P 526 O 107 P 101 P 202, P 521 P 543

BOSSCHAERTS, T. BOTELHO, H.A. BOTT, E. BOU HAYA, J.V. BOU HABIB, D.C. BOUDABOUS, R. BOUILLET, L.E.M. BOUILLET, L.M. BOUJAAMA, A. BOULET, G. BOULORIZAEDH, M. BOURATBINE, A. BOURDEAU, P. BOURDOISEAU, G. BOURREAU, E. BOUSSLIMI, N. BOUSSOFFARA, T. BOUTY, L.F. BOUTY, L.F.M. BOUVIER, G. BOWES, D.E. BOZZA, M. BOZZA, M.T. BOZZA, P.T. BRACHER, F. BRAGA, D.S.F.S. BRAGA, E.T. BRAGA, M.S. BRAGA, S.L. BRAILLARD, S. BRANCAL, H. BRANDAO FILHO, S.P. BRANDAO FILHO, S.P. BRANDAO, N.A.A. BRANDAO, R.C.F. BRANDAO, S.T. BRANNIGAN, J.A. BRAS GONCALVES, R. BRASIL, H.H. BRASIL, R. BRASOV, I.E. BRAZ, J.M. BRAZIL, R.P. BRAZUNA, J.C.M. BREITLING, R. BRENNAND, A. BRESCIANI, K.D.S.

O 342 P 798 P 334 P 725 P 183 P 908 P 434 P 135, P 242 P 067 P 654 P 452 P 591 P 355, P 656 P 484, P 790 O 221 O 221 O 273 P 790 P 153 P 499 P 502 P 403 O 265 P 118, P 136 P 446 O 051, P 013 O 275 P 399, P 414 P 162 P 054, P 056, P 686 P 143 P 143 P 533, P 816 P 636 P 596, P 642 O 293 P 659 P 850 P 906 P 789 O 009, O 128 P 609 P 219, P 293, P 296, P 330, P 335, P 585, P 608, P 614, P 617, P 650, P 940, P 941 P 613 P 902 P 821 O 152 P 159 P 209, P 211 P 160, P 955 P 518, P 607 P 261 P 057, P 065, P 746, P 818 O 296 P 384 P 101 P 402

BRETTMAN, E. BRILHANTE, A.F. BRILHANTE, M.A. BRIONES, M.R. BRITO, A.M.G. BRITO, J.F. BRITO, L.P.S. BRITO, M.E.F. BRITO, R.C.F. BRITO, R.S. BRODSKYN, C. BRODSKYN, C.I. BRUCE, K.K.M. BRUNK, B. BRUNO, F. BRUSTOLONI, Y.M. BRYCESON, A. BUENO, L.L. BUEREN, J.A. BUFFET, P. BUFFET, P.A. BULA, I. BUMB, R.A. BURCHMORE, R. BURCHMORE, R.J. BUREL, A. BURRUS, R.G. BURZA, S. BUSSOLOTTI, A.S. BUTAUD, T. BUTLER, C.D. BUTT, W.A. BYRD, J. C, J. C. MOREIRA, P.A. C. RODAS, L.A. CABEZAS, Y. CABRAL DE FRANCA, T.M. CABRAL, G. CABRAL, L. CABRERA, L. CABRERA, M. CABRERA, R. CACERE, C.R. CACERES, O.C. CADENA, H. CAETANO, R.C.S. CAFFARENA, E.R.

O 074, P 277 P 838 P 171 O 030 P 612 P 765 P 322 P 906 P 585 P 010, P 293, P 296, P 411, P 608, P 609, P 614, P 617, P 850 P 135 P 170 P 038, P 136 P 187 O 275, P 036 O 238, P 223 P 923 O 142 P 064, P 400 P 569, P 579 O 201 O 257, P 486, P 491 P 525 O 085, O 362, O 363, P 061 P 457 P 722 O 171 P 356 P 100 O 033 O 190 P 680 O 108, O 356, O 057, O 261 P 759 P 068 O 062 P 690 O 369 O 238 P 876 P 863 O 190 P 860 O 341 P 463 O 267 P 127, P 203, P 298, P 302 P 835 P 231 P 345 O 011, O 196, O 380, P 904, P 910 P 233 P 550

CAGUANO CAZA, W.A. CALABRESE, K.S. CALDART, E.T. CALDAS, I. CALDEIRA, R.D. CALDERON, W. CALEARE, A.O. CALEGARI SILVA, T.C. CALHEIROS, M.B. CALIARI, M.V. CALOGEROPOULOU, T. CALVO BADO, L. CALVO BADO, L.A. CALZADA, J. CALZADA, J.E. CAMARA, A. CAMARA, C.A. CAMARA, F.V. CAMARASA, M. CAMARASA, M.J. CAMARGO NEVES, V. CAMARGO NEVES, V.L. CAMARGO, D.G. CAMARGO, L.M.A. CAMELO, N.L. CAMERA, C.A. CAMPAGNARO, G. CAMPI AZEVEDO, A.C. CAMPILLO GIMENEZ, L. CAMPINO, L. CAMPOS SALINAS, J. CAMPOS SANTOS, E.C. CAMPOS, A.L.B. CAMPOS, D. CAMPOS, J.M. CAMPOS, M.B. CAMPOS, S.P. CAMPOS, T. CAÑADAS, K.E.B. CANAVACI, A.M.C. CAÑAVATE, C. CANAVEZ, F. CANCINO, B. CANDANEDO GUERRA, R.M.S.N. CANDLISH, A.A. CANESE, A. CANGUSSU, S.

P 918 P 139 P 192, P 507 P 713, P 715, P 716, P 787, P 826 O 180 P 864, P 867 P 535 P 625 O 037 P 885, P 891, P 894 P 886 P 888, P 889, P 890, P 897 P 316 O 310 O 197, P 540 P 544 P 146, P 895 P 009 O 180 P 593 P 640 P 865, P 872 O 307 P 529 O 377 P 869 P 686 P 345 P 907 P 492 P 357, P 666 O 276 O 330 O 219, P 163, P 453, P 463, P 503, P 805, P 887 P 417 P 563 P 935 P 343 P 143 P 167, P 169, P 193, P 212, P 424 P 568 O 240 P 918 P 165 P 018, P 253 O 215, P 934 P 082 P 722 P 685 P 450 P 919 P 310

CANNONE HERGAUX, F. CANTANHEDE, L.M. CANTILLO, O. CANTINHO JUNIOR, J.J. CANTO CAVALHEIRO, M.M. CANTO CAVALHEIRO, M.M.D. CANUTO, G.A.B. CAPITANI, C.O. CAPPARELLI, F.E. CAPPELLAZZO, A.C. CAPPIELLO, S. CARANHA, L. CARAZZOLLE, M.F. CARBAJO, D. CARCELEN, J. CARDIM, L.L. CARDINOT, C.B. CARDOSO, C.R. CARDOSO, F.A. CARDOSO, F.O. CARDOSO, J.M.O. CARDOSO, L. CARDOSO, L.S. CARDOSO, M. CARDOSO, R.M. CARDOSO, T.M. CARDOZO, R.M. CARGNELUTTI, D.E. CARIDHA, D.P. CARILLO, E. CARIOCA, S.M. CARLOS, S.M. CARLSEN, E.D. CARMO, R.F. CARN, G. CARNEIRO FEIJO, F.M. CARNEIRO, A.L. CARNEIRO, C.M. CARNEIRO, D.D.M.T. CARNEIRO, G. CARNEIRO, J.C. CARNEIRO, L.A. CARNEIRO, M. CARNEIRO, M.P.D. CARNEIRO, M.W. CARNES, J. CARNIELLI, J.B.T. CAROLLO, C.A.

P 262 P 747 P 179, P 804 P 709 P 319 P 120, P 346, P 408, P 523, P 563 P 421 P 373 P 499 O 157 P 404 O 292 P 876 O 030 P 467 P 113 O 039, O 100 P 771 P 687 O 238 P 764 O 296, P 929 P 139 O 091, P 067, P 072, P 714 O 293 P 476, P 503, P 726, P 777 P 141 O 166 P 428, P 546 P 021 O 038 P 104 O 114 O 293 P 813, P 892 P 297 O 053 O 374 O 365 P 865, P 868, P 872, P 874, P 875, P 878, P 879 P 570 O 257 O 091, P 067, P 071, P 072, P 129, P 434, P 714 O 039, O 100 O 116 P 677 P 422, P 855 P 900 P 071, P 434, P 533, P 816, P 903, P 954 O 156 P 022 P 113 P 349 P 521, P 543

CARPINTERO, J. CARRASCO, N. CARREGAL, V.M. CARREGARO, V. CARREIRA, L.M. CARREIRA, V.S. CARREﾃ前, H. CARRIJO, R.S.G.G. CARRILLO, E. CARRILLO, L.M. CARRILO, E. CARSON, C. CARTER, K. CARTER, K.C. CARTER, N. CARVALHAL, D.G.F. CARVALHEIRO, M. CARVALHO ANDRADE, A. CARVALHO DANTAS, I.M. CARVALHO, A.K. CARVALHO, A.M. CARVALHO, A.S. CARVALHO, A.W.S. CARVALHO, B.M. CARVALHO, C. CARVALHO, D.M. CARVALHO, E.

P 769 O 016 P 416 O 210, O 328 P 772 P 634 P 448 P 556 P 725 O 215, O 016, O 046, P 938 O 011, O 280, P 904 P 937 O 197, P 513 P 356 O 115 P 053 P 450 O 348 P 207 O 312, O 336, P 508 P 105 P 266 P 267 P 142, P 793 O 250 P 023, P 024 P 163 P 906 P 705 P 780 O 170, P 583 O 240, O 351, P 264

CARVALHO, E.M. CARVALHO, F.A.A. CARVALHO, F.G. CARVALHO, F.S. CARVALHO, G.M.L. CARVALHO, K.I. CARVALHO, K.I.L. CARVALHO, L. CARVALHO, L.O.P. CARVALHO, L.P. CARVALHO, P. CARVALHO, P.S.L. CARVALHO, R.M.A. CARVALHO, R.S. CARVALHO, S. CARVALHO, S.F.G. CARVALHO, S.M.S. CARVALHO, S.S.

O 233, O 277, O 050, P 128, P 204 O 241, O 272, P 294 P 480 O 278, O 166, O 169, P 021, P 023, P 024, P 087, P 134, P 141, P 337, P 423, P 527, P 590, P 785 P 133 P 689 P 906 P 928 P 655 P 800 P 193 P 142 O 240, O 278 P 192 P 204 P 294 O 241, O 272, O 087 P 023 P 243 P 362 P 721 O 170, P 583 O 182, P 185 O 070 P 496 P 953 P 261, P 456

CARVALHO, T.B.F. CARVALHO, Y. CASANOVA, C. CASARIL, A.E. CASTAGNINO, M. CASTANYS, S. CASTELAO, K.G. CASTELLI, G. CASTELLS, X. CASTELLUCCI, L. CASTELO BRANCO, R.C. CASTIGLIONI, P. CASTILHO MARTINS, E.A. CASTILHO, R.O. CASTILHO, T.M. CASTILHO, W.S. CASTILLEJO, S. CASTRO BORGES, W. CASTRO OLIVEIRA, R. CASTRO, A.A.P. CASTRO, E.A. CASTRO, E.V. CASTRO, H. CASTRO, J.M. CASTRO, J.S.M. CASTRO, L. CASTRO, L.G.M. CASTRO, M.C.A.B. CASTRO, N. CASTRO, R. CASTRO, R.A.O. CASTRO, R.A.O.E. CASTRO, R.O. CASTROVINCI, P. CASTROVINCI, P.A. CATARINO, C.M. CATCHPOLE, B. CATHERINE, R. CAUAS, E.M.C. CAUMES, E. CAVALCANTE DE OLIVEIRA, A. CAVALCANTE SILVA, L.H.A. CAVALCANTE, G.G. CAVALCANTE, T.V. CAVALCANTI, A.S. CAVALCANTI, M.P. CAVALCANTI, S.M.M. CAYLA, M.

P 209, P 211 P 098 P 746, P 748, P 841 P 057, P 823, P 832, P 846, P 893 P 707 P 413, P 417 P 601 P 064, P 400 P 944 P 133, P 264 P 436, P 729, P 730, P 732, P 733 O 035, O 052, P 112 P 373 O 129 P 376 P 520 P 180 P 080, P 718 P 944 P 126, P 143 P 197 P 856 O 096 P 236 P 098, P 441, P 444, P 742 P 761 O 335 O 070, O 188, O 059, P 262, P 471 P 728, P 834 P 817 P 283 P 341 P 411, P 566 P 429 P 311, P 317 P 509 P 126 O 082 P 807 O 320 O 161 P 738 O 213 O 052 P 616 P 457 P 198 P 571 P 864, P 867 P 856 O 184 P 335, P 906 P 293 O 205

CEBALLOS, A.L. CEDEÑO, D.L. CELES, F. CELES, F.S. CELESTE, B.J. CELESTE, J.L. CELESTE, J.L.L. CEN, S. CERDA, R.M.G. CERIZE, N.P. CESETTI, M.V. CEZAR, T.L. CHAARA, D. CHAGAS, A.P. CHAGAS, B.D. CHAKRABARTI, S. CHAKRAVARATY, J. CHAKRAVARTY, J. CHAKROUN, A. CHAKROUN, A.S. CHALGHAF, B. CHAMAKH AYARI, R.C. CHANCE, M.L. CHANDRAWANSA, P.H. CHANG, K.P. CHAOUCH, M. CHAPPUIS, F. CHARREL, R. CHATELAIN, E. CHAUDHARY, K. CHAUDHARY, R. CHAVES, C.S. CHAVES, L.F. CHAVES, S.A.M. CHAVES, S.P. CHAVEZ FUMAGALLI, M.A. CHAVEZ FUMAGALLIA, M.A. CHECA, R. CHELBI, I. CHEMKHI, J. CHEN, H. CHEN, S. CHENIK, M. CHERNI, S. CHICHARRO, C. CHILDERSTONE, M. CHIÑAZ PEREZ, A. CHIOCCOLA, V.L.P.

O 365 O 283, P 327, P 328 O 326 P 433 P 446 P 341 P 795 P 200 P 377 P 918 P 407 P 648 P 787 P 140 P 949, P 952 P 818 P 030 P 186 O 315, O 163, P 478, P 582 O 138 P 096 O 125 O 119 O 158 O 217 O 329, O 288, O 028, P 026 P 449 P 586 O 356 O 269 O 194 P 636, P 642 O 104 P 063 P 171, P 174 P 895 P 415, P 725, P 756 P 574 P 362 P 018, P 123, P 520 P 226 O 342 P 654 O 125 O 194, O 289 O 285 O 208, O 306, O 119 P 654 O 106, O 215, O 068, P 188, P 934 P 366 P 882 P 160

CHIRINOS, Y. CHIYO, L. CHLIF, S. CHOO, A.Y.L. CHOO, Y.L. CHOUDHURY, T.A. CHOUHAN, G. CHOURASIA, A. CHOWDHURY, R. CHOWDHURY, S. CHOWDHURY, T.A. CHOWDHURY, V. CHRISTODOULOU, V. CHROBAK, M. CHUSRI, S. CIANCAGLINI, P. CIARLINI, P.C. CILLI, M.L.C. CLARK, S.C. CLOKE, T. CLOS, J. CLOSS, E. CLOUTIER, S. COELHO NETO, G.T. COELHO FINAMORE, J. COELHO, A.C. COELHO, C.H. COELHO, E.A. COELHO, E.A.F. COELHO, E.F. COELHO, G. COELHO, G.M. COELHO, Z.C.B. COHEN, S. COIMBRA, E.S. COIMBRA, M. COLER, R.N. COLHONE, M.C. COLLA JACQUES, F.E. COLLAVERI, J. COLLOVATI, M. COLOMBO, F.A. CONCECAO SILVA, F. CONCEICAO SILVA, F. CONDE, C.A. CONDELO, H.S. CONRADO DOS SANTOS, W.L. CONSORTIUM, S.

P 259 P 826 O 125, P 305, P 306, P 942 O 367 P 110 O 314, P 665 P 466 O 015, P 478, P 749 P 753 P 359, P 365, P 401 O 222 P 753 O 135 O 105 P 662 P 108 P 398 P 808 P 319 O 133, P 410 O 001, O 006, O 105, O 149 O 133 O 101 P 212 O 051 P 405 P 253 P 226 P 191 P 375 P 123, P 253, P 362, P 376 P 018, P 520 O 157 P 629 P 145 P 174 O 003 P 459, P 568 P 887 O 092, O 093, O 027 P 108 P 748, P 841 P 746 P 600 P 615 P 505 O 122 O 237, P 037, P 090, P 164 P 560, P 573 P 083 P 522, P 524, P 526 O 345, P 058, P 059

CONSTANTINO, C. CONTER, C.C. CONTRERAS, M.A. CONVIT, J. COOMBS, G.H. CORBETT, C.E.P. CORBETT, C.P. CORDEIRO DA SILVA, A. CORDEIRO, J.L.P. CORDEIRO, J.P. CORDOVA, G.R. CORREA JUNIOR, J.D. CORREA OLIVEIRA, R. CORREA, I.A.A.Q. CORREA, J.R. CORREA, M. CORREA, P.R. CORREA, Z.J. CORSET, C. CORSET, M. CORTES BOITE, M. CORTES, L.M.C. CORTES, S. CORTESE, L. CORTEZ MARCOLINO, L.M. CORTEZ, A.M.C. CORTEZ, D.A.G. CORTEZ, L.R.C. CORTEZ, S.V. CORVO, M.L. CORWARE, K. CORWARE, K.D. COS, P. COSSIO, A. COSTA E SILVA, M.F. COSTA FERREIRA, M.E. COSTA FERREIRA, V.S. COSTA RIBEIRO, M.C.V. COSTA SILVA, M.F. COSTA SOUZA, B.L.S. COSTA VAL, A.P. COSTA, A.A. COSTA, A.J. COSTA, A.P. COSTA, A.S.N. COSTA, B.H. COSTA, B.L. COSTA, C.

P 713, P 716 P 554 O 252, P 899 P 951 P 797, P 909 P 053, P 450 O 033 P 142, P 146, P 167, P 169, P 212, P 291, P 793 P 193, P 422, P 424 O 330, O 372, O 293, P 079, P 476, P 519, P 726 P 696 P 699 P 918 O 081 P 242, P 533 P 588 P 556 O 361 P 065 P 424 O 133 O 133 P 711 P 065 O 219, P 163, P 463, P 503, P 805, P 887 P 048 P 354 P 897 P 889 P 848, P 885, P 886, P 890, P 894 P 871 P 421 P 848 P 871 P 955 O 312 O 133, P 410 P 437 P 452, P 460 O 099, O 258, P 737, P 859 P 069 P 738 P 166 P 236 P 738 O 276, P 643 P 208 O 116 O 186 P 765 P 788 P 759 P 764 P 685 P 758, P 764 P 191 P 304 O 349

COSTA, C.H.N. COSTA, D. COSTA, D.C. COSTA, D.L. COSTA, E.C. COSTA, E.M. COSTA, F.B. COSTA, G.F. COSTA, J. COSTA, J.F. COSTA, J.L.F. COSTA, J.M. COSTA, J.R.A. COSTA, K.F.L. COSTA, L. COSTA, L.D. COSTA, L.E. COSTA, L.H.C. COSTA, M.A. COSTA, M.D. COSTA, N.A. COSTA, P.L. COSTA, P.R.R. COSTA, R. COSTA, S.M. COSTA, T. COSTA, T.A. COSTA, T.F.R. COSTA, T.R. COSTA, V. COSTA, W.A. COTA, B.B. COTA, G.F. COTOVIO, M. COTRIM, P. COTRIM, P.C. COTTON, J. COTTON, J.A. COUBEU, S.B. COURA VITAL, W. COURTENAY, D.A.O. COURTENAY, O. COUTINHO COSTA VELOSO, O.M. COUTINHO SILVA, R.

P 323 P 405 O 017, O 198, O 354, O 358, O 322, O 069, P 052, P 060, P 195, P 275, P 276, P 279, P 280, P 281, P 290, P 319, P 561, P 613, P 639, P 645, P 689, P 775 O 328 P 138 O 017, O 198, O 069, P 052, P 060, P 195, P 275, P 279, P 280, P 281, P 613, P 645 P 290 P 021 P 202, P 521 P 436, P 729, P 730, P 732, P 733 P 685 P 181 O 291, P 187, P 343 P 548 P 952 P 132 P 813, P 892 P 865, P 868, P 872, P 875 P 375 P 268 O 157, P 018, P 123, P 226, P 362 P 376 P 780 P 907 P 660 P 545 O 231 P 650, P 659, P 782 P 408, P 563 O 240 P 204 P 696 P 222 P 803, P 807 P 263 P 362 O 070 P 699 P 516 P 228 O 224, P 358 P 476, P 726 P 472 P 577 P 637 O 140, O 141, O 143, O 344, P 053, P 672 O 298 P 441, P 444, P 742 P 071, P 434, P 533, P 816 P 906 O 041, O 042, O 197, P 513, P 540 P 321, P 322 P 555

COUTINHO, O.M.V.C. COUTINHO, S.G. COUTO, L.S. COVAS, C.J.F. CRAVO, P. CREPALDI, P.H. CRISANTE, G. CRISTAL, J. CRISTAL, J.R. CRISTALDO, G. CRISTIANO, M.L. CRISTOVAO, E.C. CRISTOVAO, J.M. CROFT, S.L. CRUZ, A. CRUZ, A.K. CRUZ, A.S. CRUZ, E. CRUZ, I. CRUZ, J. CRUZ, K.P. CRUZ, M. CRUZ, M.D.S.P. CRUZ, M.D.S.P.E. CRUZ, M.E. CRUZ, M.E.M. CRUZ, M.S. CRUZ, M.T. CRUZ, T. CUENCA, M. CUERVO ESCOBAR, S. CUERVO, P. CUIN, A. CUISINIER, A. CULL, B. CUMAN, R.K.N. CUNHA DO NASCIMENTO, M.T. CUNHA JUNIOR, E.F. CUNHA NETO, E. CUNHA, C.F. CUNHA, G.D. CUNHA, G.M.R. CUNHA, G.V.D. CUNHA, J. CUNHA, J.C. CUNHA, J.C.L. CUNHA, J.L.

P 572, P 619 P 269 P 215, P 235 P 228 P 238 P 303 P 103 P 836 P 024 P 187, P 283 P 838 P 463 P 236, P 858 P 805 O 219, P 887 O 334 O 348 P 198, P 312 P 729, P 730 O 182, P 508 O 106, O 215, O 259, O 068, P 188, P 934 P 310 P 326 P 074 P 245 P 468, P 689, P 770 P 720, P 936 O 336 O 312, P 471 P 618 O 372 P 262 P 298, P 302 O 325 O 325, P 124, P 243 P 459 O 212, O 214, O 215, P 068 O 033 P 233 O 301, P 420 P 408 P 563 P 033 P 150, P 237 P 876 P 069 P 815 O 046, O 372, O 293 P 038 P 880 P 813, P 815, P 892 P 876

CUNHA, J.L.R. CUNHA, J.M. CUNHA, P.N.A. CUNNINGHAM, J. CUPOLILLO, E. CUSTODIO, E. CUSTODIO, M.G.F. CUTOLO, A.A. CUYPERS, B. D, A.K. DA CONCEICAO, W.M. DA COSTA LIMA, T.D. DA COSTA SILVA, T.A. DA COSTA, A.G. DA COSTA, A.P. DA COSTA, C.M. DA COSTA, E.C. DA COSTA, J.B.N. DA COSTA, L. DA COSTA, L.B. DA COSTA, L.D. DA COSTA, R.V. DA COSTA, S.C.G. DA CRUZ, A.M. DA CRUZ, N.C.A. DA CRUZ, P.F. DA CUNHA, J.P. DA FONSECA, A.C. DA GRACA, G.C. DA MATTA, C.B.B. DA MOTTA, J.C. DA MOTTA, J.O.C. DA ROCHA, D.R. DA ROCHA, W.D. DA SILVA JUNIOR, V.A. DA SILVA, A. DA SILVA, A.D. DA SILVA, A.E. DA SILVA, A.F. DA SILVA, A.J.M. DA SILVA, C.V. DA SILVA, D.T. DA SILVA, E.D. DA SILVA, E.O. DA SILVA, E.S. DA SILVA, F.J. DA SILVA, F.M.M.

P 244 P 223 P 345 P 361 O 136, O 137, O 139, O 184, P 073, P 082, P 119, P 320, P 568, P 711, P 866, P 921, P 949 P 934 P 747 P 601, P 631, P 911, P 912, P 928, P 957 P 384, P 385, P 386 O 023 P 609 O 103 P 367 P 258 P 155 P 956 P 543 P 528 P 829 P 834 P 635 P 740 P 139 P 150 P 432 P 224 P 282 P 735 P 320 P 528, P 571, P 593, P 594 O 083 P 428 P 409 P 226 P 018 P 640 P 581, P 860 P 459 P 440, P 528, P 594 P 864, P 867 P 408 P 563 P 161, P 229, P 247, P 252, P 511, P 512 P 863 P 539 P 522 P 782 P 708 O 009 P 659 P 902

DA SILVA, G.O. DA SILVA, G.P. DA SILVA, I.S. DA SILVA, J.C. DA SILVA, J.R. DA SILVA, J.S. DA SILVA, K.L.O. DA SILVA, L.C. DA SILVA, L.C.O. DA SILVA, L.H.P. DA SILVA, L.V. DA SILVA, M.F.L. DA SILVA, M.R. DA SILVA, S.M. DA SILVA, T.A.M. DA SILVA, V.M.G. DA SILVEIRA NETO, M.M. DA SILVEIRA PERES, L.L. DA SILVEIRA, J.F. DA CRUZ, A.M. DA SILVA, S.A. DABIRI, S. DABIRIAN, S. DADO, D. DAGGER, F. DAGNE, D.A. DAIFALLA, N.S. DALAGRANA, L. DALIGAULT, F. DAMAJ, S. DAMASCENA, N.P. DAMASCENO, A.R. DAMASCENO, L. DAMASCENO, L.M. DAN GOOR, M.E. DANCKAERT, A. DANDREA, A. DANTAS PEREIRA, P.D. DANTAS TORRES, F. DARABI, M.H. DARDONVILLE, C. DARGHOUTH, M. DARVISHI, M. DAS, A. DAS, M.L.

O 156 P 678 P 729, P 730 O 069, P 052, P 275 P 581, P 860 P 500 P 253 P 688 P 676 P 864 P 420 O 322 O 048, O 129, P 465, P 789 P 240 P 239 O 082, P 129, P 606, P 839 P 903 P 282 P 633 P 893 O 030 P 228, P 269 O 233, O 234, P 215, P 217, P 234, P 235, P 237, P 238, P 241, P 447, P 551, P 569 P 124 P 591, P 701 O 239 O 342 P 429, P 494 O 094 O 355 O 117 P 738 O 190 P 684 P 576 P 864, P 867 P 213 P 226 P 191, P 537 O 032 O 284 P 050 O 009 O 009, O 010, O 128, P 219, P 330, P 335, P 650, P 940, P 941 O 195, O 127 P 461 P 586 O 195, O 127 O 154 O 111, O 067

DAS, P. DAS, P.K. DAS, S. DAS, V. DAS, V.N. DAS,, S. DASH, A.P. DASKALAKI, A. DAVID, C.C. DAVID, J.R. DAVIES, M. DAZA, J.A. DE ALMEIDA LEITE, J.R.S. DE ALMEIDA, A.B. DE ALMEIDA, A.C.O. DE ALMEIDA, A.F. DE ALMEIDA, A.P. DE ALMEIDA, A.P.F. DE ALMEIDA, E.L. DE ALMEIDA, L. DE ALMEIDA, M.C.C. DE ALMEIDA, R.P. DE ALMEIDA, T.M. DE AMORIM, I.F.G. DE ANDRADE PINTO, P.C.E. DE ANDRADE, A.J. DE ARAUJO, C.N. DE ARAUJO, E.A.C. DE ARAUJO, J.M.P. DE ARAUJO, K.C.L. DE ARAUJO, V.C. DE ARAUJO, V.C.P. DE ARRUDA, C.C.P. DE AZEVEDO, A.C.R. DE AZEVEDO, A.F. DE AZEVEDO, R.B. DE BARROS, N.B. DE BRITO, C.F.A. DE BRITO, M.E.F. DE BRITO, R.F. DE CAMARGO, R.A. DE CARVALHO, A.T. DE CARVALHO, G.Q. DE CARVALHO, G.S. DE CARVALHO, L. DE CARVALHO, L.C.P. DE CARVALHO, L.M.

P 063 O 108, O 056, O 057, O 061, O 256, O 261, P 599, P 603 O 300 P 041, P 042, P 043, P 381 O 235, P 088 O 061 O 057 O 061 O 089 P 753 P 059 P 640 O 271 O 347 O 280, P 589 P 195, P 561 P 695, P 712 O 234 P 010, P 205, P 220, P 265, P 566 P 265 O 184 P 850 P 633 P 538, P 553 P 526 O 151, P 261, P 504, P 576 P 568 P 010, P 205, P 220, P 566 P 316 P 097 O 007, P 649 P 414 P 345 P 876 P 229 P 202 P 543 P 543 P 948 P 504 P 500, P 510, P 556 P 325 P 225 P 330, P 566, P 633 P 242 O 168, P 435 O 091, P 496 P 136 P 459 O 030 P 020, P 182, P 271, P 282, P 522 P 490

DE CARVALHO, M.R. DE CARVALHO, T.G. DE CASTRO FREITAS, T.M. DE CASTRO NETO, A.L. DE CASTRO, E.M. DE CASTRO, M.C.A.B. DE CELIS, H.A. DE CROP, M. DE DONCKER, S. DE FARIAS, D.M. DE FIGUEIREDO, H.R. DE FIGUEIREDO, L.C. DE FRANCA MENDES, F.G. DE FRANCA, S.H.F. DE FREITAS, E.O. DE FREITAS, J.C. DE FREITAS, L.A. DE JESUS, A.C. DE JESUS, A.R. DE JESUS, E.A. DE JESUS, J.B. DE JESUS, R.C.S. DE LA FUENTE, S. DE LA GUARDIA, C.I. DE LIMA COSTA, K.F. DE LIMA, B.D. DE LIMA, D.A. DE LIMA, H. DE LIMA, I.D. DE LIMA, V.M.F. DE LOS SANTOS, M.B. DE LUCA, P.M. DE LUCCA, H.R.L.R. DE LUCENA, N.C. DE MACEDO, L.B. DE MACEDO, M.S.F. DE MEDEIROS, J.F. DE MELLO, M.F. DE MELO NETO, O.P. DE MELO, E.V. DE MELO, G.D. DE MELO, I.M. DE MELO, M.F. DE MELO, S.D. DE MENDONCA, I.L. DE MENEZES, J.P.B. DE MIRANDA, M.B. DE MORAIS, I.B.

P 850 P 860 P 321 P 271, P 592 P 225 P 010, P 205, P 220, P 265 P 227 O 262 P 053, P 383 P 864 P 823, P 832 P 060 P 860 P 782 P 470 P 713, P 715 P 639 P 562 O 209, O 151, P 576 P 729, P 730 P 124 P 855 P 769 P 009 P 874, P 878, P 879 P 428, P 688 P 695, P 698 O 259, P 797, P 909, P 914 P 896 O 185, P 396, P 398, P 688 P 835 P 210 P 489 P 633 P 868, P 875 P 878 P 222 P 920 O 156 O 103 P 592 P 270, P 271 O 209, P 740 P 049, P 196 P 492 P 099 P 782 O 322 P 106 P 240 P 167

DE MORAIS, O.O. DE MORAIS, R. DE MORAIS, R.C.S. DE MOURA, F.K. DE MOURA, S.A.L. DE MOURA, T.C.M. DE MOURA, T.R. DE MUYLDER, G. DE OLIVEIRA E CRUZ, D.M. DE OLIVEIRA NETO, M.P. DE OLIVEIRA SILVA, E.P. DE OLIVEIRA, A.F.J. DE OLIVEIRA, A.H.C. DE OLIVEIRA, A.J.F. DE OLIVEIRA, A.L.L. DE OLIVEIRA, A.N. DE OLIVEIRA, A.P. DE OLIVEIRA, B.C. DE OLIVEIRA, B.M.N. DE OLIVEIRA, C.I. DE OLIVEIRA, D.E. DE OLIVEIRA, E.F. DE OLIVEIRA, F.A. DE OLIVEIRA, G.M. DE OLIVEIRA, J.H. DE OLIVEIRA, L.B. DE OLIVEIRA, M.A. DE OLIVEIRA, M.F. DE OLIVEIRA, M.Y.K. DE OLIVEIRA, R.N. DE OLIVEIRA, T.F. DE OLIVEIRA, V.C. DE OLIVEIRA, W.A. DE PADUA MILAGRES, F.A. DE PAIVA CAVALCANTI, M. DE PAIVA, R.M. DE PAULA, C.D.R. DE PAULA, N.A. DE PAULA, R.C. DE RESENDE, D.M. DE SA, J.C. DE SA, W.T. DE SIMONE, S.G. DE SOUSA, A.C. DE SOUSA, C.B. DE SOUSA, F.D. DE SOUSA, L.C.

P 648 P 867 P 330, P 585, P 650 P 876 P 240 P 240 O 151, P 250, P 261, P 456, P 504, P 576, P 740 O 285 P 956 O 234 P 354 P 924 P 557 P 925 P 569 P 543 P 864 P 205, P 220, P 566 P 205, P 220, P 265, P 566 P 182 O 250, P 187 O 087, P 433 P 223 O 238, O 326, P 022, P 024, P 446, P 548 P 574 P 823, P 832 P 676 O 151, P 576 P 860 P 681 P 616 P 354 O 304 P 217 P 492 P 158 P 012 P 832 P 321, P 322 P 219, P 330, P 585, P 650 O 350 P 530 P 201 P 546 P 606, P 839 P 143 P 626 P 886 P 890, P 897 P 168 P 673, P 674 P 163 P 880 P 097, P 815

DE SOUSA, L.L.F. DE SOUSA, M.D.G. DE SOUSA, M.R. DE SOUZA, B.M. DE SOUZA, C.A.C. DE SOUZA, C.C. DE SOUZA, C.L. DE SOUZA, C.M. DE SOUZA, C.S.F. DE SOUZA, E.B. DE SOUZA, F.A. DE SOUZA, J.V. DE SOUZA, M.A. DE SOUZA, M.B. DE SOUZA, N.A. DE SOUZA, O.M.F. DE SOUZA, T.L. DE SOUZA, W. DE VRIES, H. DE VRIES, H.J. DE VRIES, P.J. DE LUCA, P.M. DEACON, A. DEACON, A.D. DEBNATH, S. DEBONI, S.C. DEBORAH, D. DEBRABANT, A. DEBUS, A. DECUYPERE, S. DEDET, J. DEDET, J.P. DEEP, D.K. DEGROSSOLI, A. DEKONING, H. DEL VALLE, S.G. DELGADO, M. DELGADO, O. DELLAGI, K. DELPUTTE, P. DEMANT, P. DEMICHELI, C. DEMICHELI, C.P. DEMIR, M. DEMUYLDER, G. DEN BOER, M. DEN BOER, M.L. DEOTI, B.

P 892 P 436, P 732, P 733 O 224, P 358 P 108 P 782 P 066 P 782 P 735 P 139, P 192, P 415, P 865, P 868, P 872, P 875 P 231 P 139 P 240 P 511 P 695, P 698, P 712, P 824 P 097, P 761 O 044 P 695, P 712 O 310, P 103, P 352, P 399, P 414, P 432 O 098 P 151 O 078 O 274, P 005 O 279 P 111 P 333 P 870 O 297 O 297 O 132 O 111 O 135, O 014, O 221, P 140, P 905 P 098, P 908 P 364 P 168 P 101 P 367 P 859 P 417 O 259, P 542 P 058, P 059 P 452 O 049, O 086, O 260 O 311, P 393, P 606, P 839 O 081, O 082 P 272 O 306 O 203, O 023 O 202 P 309

DEPAQUIT, J. DERBALI, M. DESCHACHT, M. DESCHUTTER, J. DESCOTEAUX, A. DESEWU, K. DESJARDINS, M. DESJEUX, P. DESOTI, V.C. DESPONDS, C. DEY, R. DHAHRI, B. DHAKAL, S.S. DHALARIA, P. DHALIA, R. DHOM LEMOS, L. DI MARCO LO PRESTI, V. DI MUCCIO, T. DI BLASI, T. DIAO, L.Y. DIAS DE MELO, G.D. DIAS TEIXEIRA, K.L. DIAS, E.S. DIAS, F.C. DIAS, F.E.F. DIAS, G.V. DIAS, M.H.F. DIAS, R. DIAS, R.C.F. DIAS, S. DIAS, S.G. DIAS, T.L.M.F. DIAZ, B.L. DIAZ, E. DIAZ, N.L. DIAZ, R. DIETZE, R. DIKOLLI, E. DILERMANDO, J.A. DILGER, E. DILL, D.C.S. DILLON, R. DILLON, R.J. DINIS OLIVEIRA, R.J. DINIZ ATAYDE, V. DINIZ, A.G. DINIZ, M.M.C.S.L. DIOGO, E.B.

O 221 P 654 P 452 P 825 O 031, O 034, O 324, O 051, P 227 P 946 O 324 O 019 P 625 P 112, P 116 O 026 P 908 O 111 O 109, O 021 P 020 P 486 P 050 O 232, O 123, O 292, P 604 O 246 P 518, P 607 P 032 P 155 P 821 P 788 P 214, P 567, P 754, P 758, P 759, P 764, P 765, P 954 P 557 P 856 P 880 P 292 P 615 P 716, P 826, P 829 P 178 P 216 P 440, P 594 P 170 P 017, P 338 P 046 P 025 P 254 P 047 P 146 P 769 P 349 P 295 P 735 O 041 P 080, P 718 O 332 O 194 O 330 O 029 P 294 P 861 P 492

DIONISIO, E. DIONISIO, L. DIXT, U.G. DO CANTO CAVALHEIRO, M.M. DO CARMO, J.P.M. DO NASCIMENTO, L.A. DO NASCIMENTO, M.B. DO NASCIMENTO, P.P. DO VALLE, T.Z. DO VALLE MATTA, M. DOBSON, D.E. DOEHL, J. DOMMANGET, N. DOMONT, G. DOMONT, G.B. DONELSON, J.E. DONG, R. DORES, T.L. DORLO, T.P. DORO, B. DORO, D.H. DORTA, M.C.L. DORTA, M.L. DORVAL, M.E.C. DORVAL, M.E.M.C. DOS ANJOS, K.F.L. DOS ANJOS, M.S. DOS REIS, A. DOS REIS, A.S. DOS REIS, F.C.G. DOS SANTOS, A.P.A. DOS SANTOS, B.N. DOS SANTOS, E.C.T. DOS SANTOS, F.R.B. DOS SANTOS, G.R.R.M. DOS SANTOS, K.M. DOS SANTOS, L.D. DOS SANTOS, L.M.G. DOS SANTOS, M.G. DOS SANTOS, M.H. DOS SANTOS, M.S. DOS SANTOS, N.F.B. DOS SANTOS, P.C.F. DOS SANTOS, P.L. DOS SANTOS, T.V. DOS SANTOS, W.L. DOS REIS JUNIOR, C.A.

P 369 P 163 P 013 P 427 O 291 P 860 P 271 P 255 P 139 O 180 O 142, O 351 O 251 O 221 P 243, P 474 P 040 O 302 P 094 P 210 O 078, O 111 O 115, P 356 P 165 P 613 P 404 P 131, P 251, P 283, P 479, P 569, P 579, P 801 P 838, P 851, P 893 P 638, P 644 P 020 P 375 P 907 P 263 P 344, P 345 O 136 P 427 P 883 O 037 P 893 P 108 P 949 O 129, P 081 P 538, P 553 P 523 P 427 P 657 P 511 O 151, P 250, P 261, P 576, P 740 P 097, P 649, P 902 P 223 P 069

DOS SANTOS, W.L. DOS SANTOS, W.L.C. DOSSET, M. DOURADO, F.V.G. DOUSTDARY, F. DOWNING, T. DOWSLEY FILHO, ,.M.A.S. DRAGAS, S. DRAHOTA, J. DRIEMEIER, M. DRINI, S. DRUMOND, K.O. DRUZIAN, A.F. DUAILIBE, R.F. DUARTE, A. DUARTE, M.C. DUARTE, M.I.S. DUBE, A. DUBOVSKY, J. Dujardin , J.

DUJARDIN, J. DUJARDIN, J.C. DUMAIDI, K. DUMETZ, F. DUNCAN, R. DUNN WALTERS, D. DUPANOVIC, B. DUPLAY, P. DUPLESSIS, C. DUQUE, M. DUQUE, S.M. DUREAU, R. DURVASULA, R. DUTHIE, M. DUTHIE, M.S. DUTRA, J. DUTRA, J.L.L. DUTRA, J.M. DUTRA, W.O. DUVAL, R. DUZ, A.L.C. DVORAK, V. E FERREIRA, R.C. E SILVA, A.A.S. E SILVA, J.S.F. E SILVA, R.P.

P 282 O 181, O 183, P 076, P 077, P 207, P 391, P 639, P 719 O 118 P 939 O 025 O 140, O 141, O 143, O 298, O 344, P 053, P 672 P 616 O 013 O 244 P 579 O 208 O 198 P 579, P 851 P 783, P 784, P 819 O 220, O 319, P 772, P 786 P 226 O 157, P 018, P 123, P 362, P 376, P 520 P 639 O 089, O 120, P 041, P 042, P 043, P 381, P 388, P 514, P 582 O 369 O 344 O 002, O 140, O 143, O 078, O 148, O 285, O 315, O 351, O 357, O 254, O 267, P 053, P 062, P 144, P 348, P 382, P 383, P 384, P 385, P 386, P 438, P 672 P 773 O 235, O 111, O 113, P 088, P 363, P 364, P 401, P 679 O 243 O 002 O 026 P 077 O 013, P 426 P 227 P 946 O 280 P 328 O 190 O 192, O 247 P 250, P 615 P 576 O 092, O 093, O 027 O 266 O 209 P 950 P 504 P 840 O 277, O 050 P 555 P 714 O 216, O 248, O 249 O 030 P 319 O 083 P 330, P 650

EATON, P. ECHEVERRI, L.F. EDGEL, K.A. EFSTATHIOU, A. EGEA, A.C. EGURROLA, J.A. EIDE, D. EJARA, E.D. EJGHAL, R. EJOV, M. EL AHMADI, Z. EL HASSAN, A.M. EL HESSNI, A. EL SAFI, S. EL SAFI, S. ELAMINE, W. ELEUTERIO, C. ELIAS, C.R. ELKHOURY, A.N. ELKHOURY, A.N.S.M. ELLIS, S. ELLIS, S.J. ELNAIEM, D. ELNAIEM, D.A. EMPKE, A. EMPKE, A.L.T. ENGEL, J.C. ENTRINGER, P.F. EQUBAL, A. ERDEM, H.A. EREN, R.O. EREQAT, S. ERMIS, Ö.V. EROL, N. ERTABAKLAR, H. ES SETTE, N. ESCACENA, C. ESCH, K. ESCOBAR, P. ESLAMI, G. ESPADA, C.R. ESPINOSA, D. ESPINOZA, J.R. ESPIR, T.T. ESSID, A. ESTAQUIER, J. ESTECHE, M.V. ESTEVAM, L.M.

P 561 P 327 O 351 P 680, P 835 P 372, P 406 P 787 O 164 O 070 O 262, O 263 P 015, P 016 O 094 O 363 O 343 P 015 O 254 O 265 O 138 O 312 P 658, P 767 P 489 O 380 O 040, O 223 O 262, P 366, P 627 O 356 O 075, O 297 O 218, O 020 P 873 P 745 O 285 P 166 P 599, P 603 P 287 O 052 O 035 O 290, P 684 O 321 O 321 O 295, O 321 O 253 P 769 P 884 P 497, P 500, P 510, P 556, P 560, P 573 P 001, P 621 P 404 O 139 P 245 P 258 P 916 O 330, O 372 P 628 P 703

ESTEVES, A. ESTIRADO, A. ESTRADA, M. ETHIER, C. EVANGELISTA, J.S.A.M. EVANGELISTA, M.G. EVERS, F. EWALD, M.P.C. EZZIKOURI, S. F. CHIERICI, N. FABER, D. FABIANO OLIVEIRA, L. FABIENNE, T. FABRA, M. FADUL, M.R. FAGUNDES SILVA, G.A. FAGUNDES, A. FAIOES, V.S. FAIRLAMB, A.H. FAIVRE CHARMOY, M. FAIZ, A. FAIZ, M.A. FAKIOLA, M. FALCAO, C.A.B. FALCAO, C.B. FALCAO, M.J.C. FALCAO, S.A.C. FALCHETTI, F. FARACO, A.A.G. FARAUT, F. FARIA, A. FARIA, A.C. FARIA, A.R. FARIA, D. FARIA, J.V. FARIA, K.F. FARIA, S. FARIAS ROTONDANO, T.E. FARIAS LUZ, N. FARIAS, J.C. FARINHA, A. FASAHAT, M. FASEL, N. FATHALLAH MILI, A. FATTORE, G. FAUCHER, B. FAUSTINO, S.K.M. FEIJENS, P.

O 336, P 508 P 769 P 686, P 852 P 336 P 502 P 472 P 713 P 829 P 016 P 863 P 332 O 159 O 326 P 113 P 952 P 234, P 238 O 122, P 824 P 408, P 427 O 153 O 297 O 109 P 366 O 343, O 352, P 133 P 547 P 407 P 622 O 326 P 845 P 520 O 014 P 309 P 069 P 537 O 277 P 523 P 807 P 753 P 842 P 118 O 198 P 163 P 621 O 035, O 074, O 052, O 351, P 112, P 115, P 116, P 218 P 501 P 825 O 014 O 322 P 460

FEIO, M.J.F. FELGER, I. FELICIANGELI, M.D. FENDRICH, R.C. FEREGUETTI, T.O. FERGUSON, M.A. FERJANI, B. FERMIN, Z. FERNANADEZ, M.S. FERNANDES, A.B. FERNANDES, A.C.S. FERNANDES, A.M.M.M. FERNANDES, A.P. FERNANDES, A.P.S. FERNANDES, A.P.S.M. FERNANDES, C.E.S. FERNANDES, C.J. FERNANDES, F.R. FERNANDES, J.R.M. FERNANDES, J.S. FERNANDES, L.S. FERNANDES, M.A. FERNANDES, M.F. FERNANDES, M.Z. FERNANDES, R.M. FERNANDES, R.P.M. FERNANDES, W.D. FERNANDEZ REYES, M. FERNANDEZ, A. FERNANDEZ, J. FERNANDEZ, J.L. FERNANDEZ, M.S. FERNANDEZ, R. FERNANDO, F.S. FERRARI, H. FERRARI, H.F. FERRARINI, S.R. FERRAZ, R. FERREIRA DA COSTA, E.S. FERREIRA SOARES, M.C. FERREIRA, A.L. FERREIRA, A.L.C. FERREIRA, A.M.T. FERREIRA, A.S. FERREIRA, C. FERREIRA, D.D. FERREIRA, E.C. FERREIRA, E.M.

P 561 O 255, P 061 O 259, P 910 P 826 O 224 O 075 P 958 P 040, P 379, P 380, P 443 P 806 P 720 P 246 P 678, P 865, P 868, P 872, P 874, P 875, P 878, P 879 P 018, P 165, P 226, P 253, P 375 P 145 P 191, P 214 P 057 P 754 P 393 O 311 P 545 P 141 P 179, P 804, P 811 P 847 O 170, P 583 P 504 P 283, P 893 P 278 P 481 P 504 P 893 P 475 P 914 P 835 P 008 O 045, O 095, O 097, P 778 P 680 P 190 P 056 P 634 P 448 P 559 P 237 O 096 P 158 P 818 P 416 P 552 P 057 P 325 P 378, P 420 P 370 P 788 P 731 P 795 P 703

FERREIRA, F. FERREIRA, F.C. FERREIRA, F.M. FERREIRA, G.E.M. FERREIRA, G.M. FERREIRA, I.G. FERREIRA, I.V.G. FERREIRA, J.B. FERREIRA, J.H.L. FERREIRA, J.L.P. FERREIRA, J.O. FERREIRA, L.A.M. FERREIRA, L.T. FERREIRA, P.A. FERREIRA, R.A. FERREIRA, R.G.M. FERREIRA, S.A. FERREIRA, T.R. FERREIRA, V.F. FERREIRA, W. FERREIRA, W.A. FERREIRO TERCEIRO, B.R. FERRER, J. FERRER, L. FERRERA, W. FERRIERES, V. FERRO, C. FERRO, M.C. FERRUA, B. FIALHO JUNIOR, L.C. FIECK, A. FIERRO, M.N.A. FIGUEIRA, C.P. FIGUEIRA, L.P. FIGUEIREDO, F.B. FIGUEIREDO, H.R. FIGUEIREDO, J.K. FIGUEIREDO, L.C. FIGUEIREDO, M. FIGUEIREDO, M.M. FIGUEIREDO, R.C.B.Q. FIGUEIREDO, S.G. FIGUEIREDO, W.M.E. FIGUEREDO, L.A. FIGUEREIDO, J.R. FILHO, A.A.P. FILHO, C.J.L.B. FILHO, D.M.

P 937, P 938 P 956 P 509 P 126 P 866 P 073, P 082, P 320 P 119 O 136 P 885 P 888, P 889, P 891, P 894 P 897 P 886, P 890 P 788 P 350 P 445 P 210 P 393 O 116 P 587 P 007 O 276 P 316 P 532 P 747 O 294, P 416 P 198, P 312 P 492 P 409 O 311 P 393 P 090 P 113 O 110 O 279 O 190 P 859 O 008 P 430 P 066 O 247 P 918 P 207 P 258 O 296, P 415, P 695, P 698, P 712, P 929 P 846 P 857 P 276 P 309, P 311, P 629 P 316 P 640 P 638, P 644 P 349 P 034 P 171, P 174 P 219 P 622 P 673, P 674, P 675, P 734, P 783, P 784, P 819 P 712 P 498

FILHO, E.P.S. FILHO, J.D.A. FILHO, J.D.S. FILHO, L.X. FILHO, O.A.M. FILHO, S.P. FILHO, V.P. FINKELSTEIN, L.C. FIORENTINO, E. FISA, R. FISCHER, L. FISCHER, M.R. FISHER, A. FITZPATRICK, S. FIUZA, J. FIUZA, V.O. FLOETER WINTER, L.M. FLOHE, L. FLORENTINO, P.V. FLORES LOPEZ, E.J. FLOREZ, A. FLOREZ, M.F. FOGLIA MANZILLO, V. FOLEY, J.E. FONSECA, A.C. FONSECA, A.F.Q. FONSECA, A.M. FONSECA, G.S. FONSECA, I.P. FONTELES, R.S. FORDELLONE, M.R.C. FORSHAW, A. FORTES DIAS, C.L. FORTIN, A. FOSTER, L.J. FOTOUHIARDEKANI, R. FRAGA, D.B.M. FRAGA, J. FRAGA, T.L. FRANCA COSTA, J. FRANCA DIAS, M.H. FRANCA SILVA, J.C. FRANCA, A.N. FRANCA, A.O. FRANCA, E.M.O. FRANCA, J.R. FRANCA, T.M.C. FRANCESQUINI, F.

P 516 P 800 P 362 P 612 P 292, P 496, P 516 P 633 P 731, P 735 P 210 O 123, O 292, P 604 O 366, P 084, P 722 O 320, P 038 O 333 P 543 O 102 O 026 P 759, P 821 P 788 O 129, P 081, P 465 O 188 O 305 P 461 O 286, P 531 O 347 O 283 O 212, O 292, P 048 P 852 P 731 P 532 P 795 P 496 O 220, O 319, P 490, P 772, P 786 P 673, P 674, P 675, P 734 P 444, P 742 O 192 P 759 O 338, O 342, P 460 O 114 P 180 P 701 O 044, P 391, P 522, P 526, P 833 O 254, P 062 P 283, P 569 P 343 P 194 P 795 P 214, P 759, P 765 P 567 P 851 P 479, P 801 P 869 P 520 P 581 P 291

FRANCISCATO, C. FRANCO, A. FRANCO, A.M.R. FRANCO, A.R. FRANCO, G.R. FREIRE, F.P. FREIRE, G.M. FREIRE, J.M. FREIRE, M.L. FREIRE, M.P. FREIRE, R.C.M. FREIRE, R.L. FREITAS, A.C.P. FREITAS, A.I. FREITAS, I.S. FREITAS, J.C. FREITAS, J.C.C. FREITAS, L.A.R. FREITAS, M.S. FREITAS, R.A. FREITAS, T.D. FREUDZON, M. FREZARD, F. FREZARD, F.J. FREZARD, F.J.G. Frezard, F.J.G. FRIESEN, J.A. FRY, L. FUCHS, J.R. FUENZALIDA, A.D. FUJIWARA, R. FUJIWARA, R.T. FUKUDA, M. FUMAGALLI, M. FUMAGALLI, M.A.C. FUMAGALLI, M.C. FUSTER, F. FUX, B. FUYA, P. FUZARI, A.A. FUZIBET, J.G. G. DE SOUSA, F.D. GABORIAUD KOLAR, N. GADELHA, S.M.S. GADISA, E. GAELLE, B. GAGO BADENAS, F. GAGO, F.

P 251 P 864 P 258, P 260 P 224, P 549 P 178 P 147 P 546 P 225 O 276 P 568 P 762 P 917 P 915 P 713, P 715, P 716, P 787, P 826, P 829 P 945 P 609 P 888, P 889, P 891, P 894 P 848, P 871, P 885, P 886, P 890, P 897 P 499 P 502, P 506 P 076 P 524 O 181 P 810 P 345 P 854 P 089 O 311, P 606, P 839 O 082, O 112, O 145, P 387, P 565 P 492 O 081 P 509 P 393 P 339 O 283 P 410 O 308 O 097 O 026 O 257, O 294, O 317, P 181, P 239, P 347, P 486, P 491 O 034 P 375 O 157 P 376 P 757, P 766, P 769 P 731 P 709 P 818 P 221 P 876 P 406 P 811 P 845, P 847 P 934 O 146, P 451 P 529 O 307, P 218, P 525

GAIDO, S.B. GALAÏ, Y. GALANTE, A. GALARDO, A.K.R. GALARZA, F. GALATI, E.A.B. GALATI, E.B. GALEANO, E. GALHARDO, J.A. GALINDO, W. GALISTEO, A.J. GALLEGO, M. GALLO, G. GALLO, L. GALUPPO, M.K. GALUTI, E. GALVAO, J.D. GALVAO, L. GALVAO, R.F.P. GALVAO, S.R. GALVEZ, R. GALVIS, F.O. GAMA, A. GAMA, M.E. GAMA, M.E.A. GAMARRO, F. GAMERO, M.D. GANGNEUX, J. GANGWAR, S. GANNAVARAM, S. GARATE, T. GARCEZ, L. GARCEZ, L.M. GARCIA BUSTOS, M.F. GARCIA HERNANDEZ, R. GARCIA PEREZ, A. GARCIA, A.L. GARCIA, B. GARCIA, E. GARCIA, F. GARCIA, G.R. GARCIA, L. GARCIA, M. GARCIA, M.H. GARCIA, M.H.O. GARCIA, R.D. GARDINASSI, L.G.A. GARG, M.

P 276 P 484 P 400, P 693 P 831 P 825 O 007, P 097, P 658, P 818, P 861, P 893, P 912, P 957 P 652, P 653, P 774, P 937, P 938 P 707 P 808 P 910 P 389 P 944, P 947 P 050 P 472 P 004, P 031 P 937, P 938 P 828 O 180 P 939 P 856 O 342 P 938 P 476 P 167 P 212 P 385, P 413, P 417 P 379, P 443 O 303, O 190, P 122 P 329 O 368, O 026 P 494 O 042, P 513 O 197 P 660, P 935, P 943, P 952 P 104 O 038 P 385 P 413 P 367 P 438 O 375 O 215 O 068 P 148, P 152 P 323 O 254 P 527 O 199 P 956 P 439 P 323 O 076

GARGANO, V. GARLAPATI, R.B. GARNAOUI, A. GARNAOUI, A.M. GAROFALO MINA, K.L. GARRIDO, L.M. GASHOUT, A. GASIMOV, E. GASPANI, S. GASPAR, M. GASPAR, M.M. GASPAROTTO, J.C. GAUDENCIO, K. GAUNA, A. GAUR, U. GAUTAM, S. GAUTUM, S. GAVA, R. GAWHARI, F. GAZZINELLI GUIMARAES, P. GAZZINELLI, R.T. GEA, S. GEBRE MICHAEL, T. GEDAMU, L. GEERTRUYDEN, J.V. GEHRKE, S.S. GELANEW, T. GEN, F. GENES, C. GENESCA, M.A.M. GENEST, P. GENNARI, S.M. GENOIS, M. GENTILE, A.G. GEORGETTI, E.D.S. GEORGIKOPOULOU, K. GERALDO, M.M. GETTI, G. GHAEDI, K. GHALIB, H. GHANEI, M. GHARBI, A. GHAWAR, W. GHEDIRA, K. GHERARDINI, P.F. GHERSI, B.M. GHIYA, B. GHODS, R.

O 187 P 651 O 119 O 208, O 236, P 039 P 918 P 747 P 724 O 094 O 203 O 182 O 312, O 336, P 471 P 236, P 849, P 858, P 862 P 659 P 380 O 302 O 206, O 207, P 086 O 299 P 160, P 601, P 631, P 911 P 450 P 486 P 165 P 035, P 145, P 191, P 213, P 537, P 562 P 104 O 248, P 029, P 750, P 751 O 117 O 012, O 316 P 482 P 308 P 313 P 218 P 392 O 107 P 685, P 809 P 336 O 038 P 856 O 310 P 156, P 172 O 279, P 111 P 299, P 300 P 610 O 318 O 125, O 289 O 125, P 922, P 942 P 059 O 346 P 680 O 171 O 318

GHOLAMI, E. GHOSH, J. GHOSH, P. GHOSH, R.S. GIACOIA GRIPP, C.B. GIAMMARRESI, M. GIDWANI, K. GIL SOARES, L.H. GIL ZAMBRANO, G.C. GILMORE, D. GIMBLET, C. GIMENEZ, A. GIMENEZ, D.S.B. GIMENEZ, G. GIOANNINI, T.L. GIRALDO, L.E.R. GIRAUD, E. GIRONA LLOBERA, E. GITHURE, G. GIUDICE, A. GIUNCHETTI, R. GIUNCHETTI, R.C. GIUSTA, C.J. GIZZARELLI, M. GODINHO, J.L.P. GODINHO, N.D.V. GODOY, E. GODOY, R.E. GOES, J.J. GOLDMAN PINKOVICH, A. GOLDSCHMIDT, M.H. GOLLOB, K.J. GOMES ALVES, A.G. GOMES SILVA, A. GOMES, A.A.D. GOMES, A.F.S. GOMES, A.H.S. GOMES, A.M.O. GOMES, A.R.F. GOMES, B. GOMES, C.C. GOMES, C.E.M. GOMES, C.M. GOMES, C.M.C. GOMES, D. GOMES, D.C. GOMES, E.V.R.

O 025 O 235, P 088 O 266 O 109, O 021 P 228 P 017 O 015, P 361, P 749 P 898 P 379, P 443 O 160, P 958 O 272 O 270 P 745 P 334 O 073 P 462 O 332, O 047, O 193, P 791 P 084, P 722 P 101 P 423, P 785 O 293 P 194 O 091, P 067, P 071, P 072, P 129, P 197, P 199, P 292, P 434, P 533, P 714 P 191 O 292 O 310, P 432 O 126, P 428 P 628 P 097 P 889, P 891 P 848, P 871, P 885, P 888, P 894, P 897 P 886, P 890 O 131 O 087 O 277, O 050 O 188 P 215, P 234, P 235, P 237, P 238, P 269, P 551 P 852 P 939 P 007 P 160, P 955 P 559 P 897 P 888, P 891 P 886, P 889 P 871, P 885, P 890 P 894 P 898 P 193, P 422, P 424 P 278 O 083, O 126, P 095, P 530, P 546, P 648 P 142, P 146, P 167, P 169, P 212, P 291 P 212 P 304 O 156 P 857

GOMES, F. GOMES, G.S. GOMES, K. GOMES, L. GOMES, L.H.M. GOMES, L.I. GOMES, M.S. GOMES, P.A.T. GOMES, R. GOMES, R.A. GOMES, S. GOMES, V.A. GOMEZ NIETO, L.C. GOMEZ OCHOA, P. GOMEZ, A. GOMEZ, C.P. GOMEZ, M. GOMEZ, M.A. GOMIDE, L.B. GONCALVES NETO, J.R. GONCALVES, A.L. GONCALVES, A.M. GONCALVES, D.D. GONCALVES, E.G.R. GONCALVES, E.L. GONCALVES, F.T. GONCALVES, G.M. GONCALVES, L.M. GONCALVES, R. GONCALVES, R.G. GONCALVES, S.A. GONCALVES, S.C. GONCALVES, V.T. GONDIM, F. GONDIM, F.C. GONGORA, R. GONTIJO, C.M.F. GONTIJO, N.F. GONTIJO, R.C. GONZAGA, W.F. GONZAGA, W.F.K.M. GONZALEZ LOMBANA, C. GONZALEZ REY, E. GONZALEZ, D. GONZALEZ, E. GONZALEZ, K. GONZALEZ, L. GONZALEZ, M.

P 850 O 220, O 319 P 950 P 759 O 264 O 319, P 490 P 260 P 358 O 276, P 903 O 257, P 491 P 541 O 160, O 161, O 320 P 440 P 563 P 703 P 113 O 110 O 365 P 806 O 270 P 825 P 227 P 157 O 258 O 147 O 029, O 099, P 089, P 100, P 737 P 731 P 767 P 761 P 158 P 713, P 715 P 212 P 755 P 867 P 108 P 471 O 312 P 660 P 921 P 945 P 335 P 330, P 585 P 650 P 528 P 600 P 407, P 575 P 100 P 812 O 276, P 225, P 762, P 798, P 800 P 239 P 606 O 317 O 276, P 225 P 156 P 172, P 175 O 272 P 417 P 766 P 757, P 766 P 146 P 248 O 108, O 261

GONZALEZ, N. GONZALEZ, Y.P. GORCII, M. GORDON, M. GORE SARAVIA, N. GOTO, H. GOTO, Y. GOTUZZO, E. GOULART, L.R. GOULD, I.T. GOULD, M. GOUNDRY, A. GOUVEIA, C. GOUZELOU, E. GOYAL, N. GOYAL, N.G. GRADONI, L. GRAFF, J.W. GRAFOV, A. GRAFOVA, I. GRAMICCIA, M. GRANO, F.G. GRAVINA, M.C.S. GRAZIANI, D. GRAZINOLLI, M. GREGGIANIN, G.F. GREKOV, I. GRIEWANK, K. GRIFFITHS, S. GRIMALDI, G. GRIMMER, G.H. GRISARD, E.C. GROGI, M. GROGL, M. GROPPI, S. GROSS, E. GRYMBERG, P. GUACHAMIN, P.E.Y. GUAN, R. GUARDO, M. GUCCIONE, J. GUDERIAN, J. GUEDES JUNIOR, J.C.O. GUEDES, C.E.S. GUEDES, H.L. GUEDES, H.L.M. GUEDES, P. GUEDES, R.P.O.

O 270 O 170, P 583 P 905, P 908 O 100 O 099, P 737, P 859 O 354, P 002, P 091, P 266, P 267, P 341, P 590 O 090, O 370, O 174, O 177, O 178 O 267 O 157, P 123 O 045, P 778, P 806 P 792 P 101 O 071 P 699, P 936 O 135 O 076, P 003, P 329 O 309 O 212, O 232, O 292, O 320 O 073, O 087 P 549 P 549 O 232, O 123, O 254, O 292, P 604, P 947 P 032, P 049, P 196 P 392 O 126, P 095, P 428 P 831 P 439 O 049 O 333 O 162 P 145 P 183 P 515 O 114 O 362, O 363, O 289, P 518, P 607 P 704 P 655 P 178 P 918 P 470 O 380 P 048 P 250, P 576 O 209, O 092, O 027, P 610 P 526 O 036, P 074, P 326, P 391 P 304 O 116, O 156, P 162, P 575 O 180 P 080, P 718

GUEDRI, E. GUEGUEN, S. GUERBOUJ, S. GUERFALI, F. GUERFALI, F.Z. GUERRA, B. GUERRA, J. GUERRA, J.A. GUERRA, J.A.O. GUERRA, J.M. GUERRA, P.V. GUERRA, R.N.M. GUERREO, R. GUERRERO, I. GUHA, R. GUIGUEN, C. GUILLEN, C. GUIMARAENS, L.H. GUIMARAES COSTA, A.B. GUIMARAES, A. GUIMARAES, E.L.A.M. GUIMARAES, L.H. GUIMARAES, L.N.A. GUIMARAES, P.N. GUIMARAES, S.S.V. GUIMARAES, T.T. GUIMARAES, V.F. GUIZANI TABBANE, L. GUIZANI, I. GUNDAMPATI, R.K.K. GUO, H. GUPTA, C.M. GUPTA, D. GUPTA, D.S. GUPTA, G. GUPTA, K. GUPTA, R. GUPTA, S. GUPTA,, R. GUSHI, L.T. GUSTIN, C. GUTERRES, S.S. GUTIEREZ, C.T. HABBOUL, Z. HAGIWARA, M.K. HAILU, A.

O 363 O 214, O 215, P 068 P 096 O 205 P 058, P 059, P 232 P 559 P 189 P 307 O 170, P 583 P 489 P 034, P 171, P 174 P 907 P 257 P 944 O 235, P 088 P 122 P 084, P 722 P 480 O 301, O 304 O 158 P 945 O 169, P 134 O 233, O 166, P 133, P 423, P 527, P 590, P 785 P 204 O 277, O 050 P 337 P 851 P 579 P 768 P 935 P 492 P 659 O 345 O 138, P 096, P 184, P 403, P 501, P 517, P 580, P 586 P 045 O 075 P 130 O 104, P 602 P 315 P 598 O 308, O 339 O 371 P 388 P 596, P 642 O 089 P 173 P 058 P 558 P 559 P 951 P 355, P 484 P 266 O 133, P 410 O 216, O 356, O 262, O 263, P 029, P 116, P 308 O 248

HAILU, W. HALABALAKI, M. HALLIDAY, A.J. HAMANO, S. HAMARSHEH, D.O.O. HAMARSHEH, O. HAMDI, S. HAMILTON, J.G.C. HANAFI BOJD, A.A. HANCOCK, R.E.W. HAND, T.W. HANNOLD, C. HAOUAS, N. HAQUE BHUIYAN, A.T.R. HAQUE, R. HARADA, S. HARALAMBOUS, C. HARBIGE, L. HARDOIM, D.J. HARIGUA, E.S. HARLEY, D. HARRABI, M. HARTKOPF, A.C.L. HARTL, D. HARTLEY, M. HARTMANN, S. HASHEMINIA, S.J. HASHIGUCHI, Y. HASKER, E. HASSAN, S.M. HASSANI, K. HASSEINE, L. HASSOUNA, N.B. HATEFI, Z. HAUYON LA TORRE, Y. HAVELKOVA, H. HAY, C.L. HAYDOCK, A. HEALY, A. HEINE, C. HEJAZI, H. HEJAZI, S.H. HEJAZI, S.H.H. HELMER CITTERICH, M. HELYNCK, O. HENARD, C.A. HENDRICKX, S.

O 133 O 216 P 487 O 158 O 084 P 455 P 092, P 684, P 724 O 253, P 015, P 016, P 092 P 746 O 127 P 377 O 121 P 518 P 140, P 905 P 366 O 084, O 109, O 021, O 176, P 366 O 179 O 135 P 111 P 192 P 403 O 018 P 305, P 306 P 131 P 019 O 035, O 052, P 112, P 115 P 320 P 299, P 300 O 038 O 012, O 015, O 316, O 357, O 055, O 163, O 067, P 749 O 109, O 021 O 029 P 221 O 208, O 236, O 306, P 039, P 153 P 621 O 250 O 086 O 049 O 053 O 348 P 461 O 166 P 301 P 620 P 300, P 913 P 621 P 299 O 346 O 284 O 053 P 452

HENRIQUES, S. HERAI, R.H. HERINGER, J.F. HERMIDA, M.D. HERMOSO, T. HERMOSO, T.J. HERNANDEZ MONTES, O. HERNANDEZ RIVERA, M.P. HERNANDEZ, C. HERNANDEZ, C.G. HERNANDEZ, L. HERNANDEZ, S. HERRERA VELIT, P. HERRNANDEZ MONTES, ,.O. HEUKELBACH, J. HIAMAMOTO, R.M. HICKMAN, M.R. HIDALGO, J.O.Q. HIDE, M. HIDER, R.C. HIGA JUNIOR, M.G. HIGINO, T.M.M. HIRAMOTO, R.I.M. HIRAMOTO, R.M. HOCHBERG, L. HOFMEYER, K. HOLDER, A.A. HOLETZ, F.B. HOLZMULLER, P. HOMBACH, A. HOMBACH, A.L. HOMEDES, J. HOMMEL, M. HORACIO, A.M.L. HOROWITZ, M.R. HORTA, M.A.P. HORTA, M.F. HORTA, M.F.M. HOSEIN, S. HOSEINI, S.G. HOSSAIN, M.S. HOSSEINI, M. HOU, Y. HOUGHTON, R. HOWARD, R.F. HOYOS, C. HOYOS, C.L. HROBARIKOVA, V.

P 453 O 030 P 551 O 183, P 207 P 380, P 443 P 474 P 040 P 595 P 595, P 882 P 600 P 781 P 462, P 803, P 807 O 342 P 757 P 245 P 882 O 043, P 739, P 853 O 296 O 114 P 918 O 366 P 482 P 851 P 644 P 631 P 395 P 489, P 540, P 601, P 767, P 857 O 289 O 092, O 027 O 152 P 020 P 159 O 006 O 001 O 110 P 366 P 567 P 293 P 708 P 102 P 178 P 646 P 620 O 084 O 318 P 055, P 697 O 194, O 289 O 093, P 610 O 266 P 485 O 038 O 249

HU, R. HU, R.V.V. HUBER, K. HUBERT, B. HUDA, M. HUEB, M. HUITEMA, A.D. HULANGAMUWA, S. HUMMER, K. HUPPERT, A. HURRELL, B.P. HURWITZ, I. HUSSAIN, M. IBRAHIM, I.C. IBRAHIM, M.E. ICEVIC, M. IGLEZIAS, S.D. IGLEZIAS, S.D.A. IGUCHI, G.U. IKEDA, F.A. IMAMURA, H. IMAMURA, R. INACIO FILHO, J.D. INACIO, R.R. INAOKA, D.K. INBAR, E. INFRAN, J.M. INFRAN, J.O.M. INIESTA, L. INIESTA, V. INSTITUT PASTEUR INTERNATIONAL NETWORK,L. IPE, A. IPE, I.M. IRETON, G. IRETON, G.C. IRISO, A. ISHIKAWA, E.A. ISHIKAWA, E.A.Y. ISHIMI, C.M. ISIKGOZ TASBAKAN, M. ISLAMUDDIN, M. ISOLA, E.L. ISOMURA, K. ITALIA, J. ITO, M.M. ITOH, M. IZIDORO, M.A.

O 098, P 458 P 151 P 399 O 221 P 796 P 158 O 078 P 670 O 160 O 216, O 065 O 326 O 192, O 247 O 124 O 051 O 343 O 013 P 160 P 955 P 823, P 832, P 846 P 395 O 002, O 143, O 298, O 344, P 672 O 168 P 346 P 090 P 164 O 179 O 072, O 297 P 823, P 846 P 832 P 084, P 722 P 113 P 663 P 667 P 667 P 610 O 266 P 757, P 766, P 769 P 422, P 424 P 060, P 275, P 810 P 893 P 117, P 287 P 466 P 334 P 313 P 450 P 747 O 173 P 538

IZQUIERDO, A.Y. JACOBUCCY, J.F. JAEGER, T. JAFARI, R. JAFFE, C. JAFFE, C.L. JAGANNADHAM, M.V. JAISANKAR, P. JAISWAL, A.K. JAISWAL,, A.K. JAKOB, U. JALALI ZAND, N. JAMIESON, S. JAMIL, K. JAMJOOM, M. JANINO, M.P.D. JANKEVICS, A. JANOTTI, A. JANSEN, H. JANUARIO, C.I. JAOUADI, K. JAQUET, D. JARA, M. JARDIM FILHO, W.R. JARDIM, A. JAROSIKOVA, T. JASKE, E.L. JASPER, L. JAVADI, A. JEAN CLAUDE, D. JEANDEL, P.Y. JEBRAN, F.A. JEDDI TEHRANI, M. JERONIMO, S. JERONIMO, S.B. JERONIMO, S.M. JERONIMO, S.M.B. JERONIMO., S.M.B. JESUS, A.M.R. JESUS, A.R. JESUS, R.S. JHA, A.N. JIA, P. JIANG, R. JIMENEZ MARCO, T. JIMENEZ RUIZ, A. JIMENEZ, A. JIMENEZ, M.

P 918 P 489 O 188 O 195, O 127, O 318, P 913 O 216 O 032, O 254 P 045 P 359 P 041, P 042, P 514 O 120, P 043 O 089 O 059 O 318 P 133, P 264 O 109 P 946 P 546 P 384 P 321, P 322, P 572 O 338, O 342, P 460 P 516 P 140, P 905, P 908 P 361 P 144, P 397, P 438 P 729, P 730 O 256, O 131 O 049 P 029 O 289 P 299, P 300, P 913 O 146, P 451 P 221 O 064 O 318 P 884 P 273, P 289 P 286 O 352 P 147 P 128, P 255, P 278 P 285, P 522 P 896 P 323 P 133, P 250, P 264, P 456 P 422, P 424 P 137 O 373 O 353 P 084, P 722 O 307, P 218 P 525, P 529 O 361, O 364, O 164 O 046 O 244, O 245 P 757, P 763, P 766

JIMENEZ, M.I. JIN HONG, W. JOCHIM, R.C. JOHNSON, A. JOJOA, J. JONES, M.A. JONES, S.L. JORGE, C.L. JORGE, L. JORGE, T.A. JOSHI, S. JOSHI,, S. JUCA, A.A. JULIAO, G.R. JUNG, J.Y. JUNIO, A. JUNIOR, C.B. JUNIOR, C.F.S. JUNIOR, E.R. JUNIOR, E.S. JUNIOR, E.T.A.M. JUNIOR, F.D. JUNIOR, J.G.C. JUNIOR, M.B. JUNIOR, M.G.H. JUNIOR, M.S.C.L. JUNIOR, S.F. JUNIOR, V.R. JURADO, D.S. JUSI, M.M.G. K. KAAK, O. KAFFELA, M. KAJILURU, L.P. KALABRESE, K.S. KALANTARI, M. KALIL, S.P. KALLAS, E.G. KAMHAWI, S. KAMMOUN REBAI, W. KAMMOUN, W.R. KANAMURA, C.T. KANASHIRO, E.H.Y. KANG, A. KANSAL, S. KAPLAN, M.H. KARAKUS, M. KARAPETYAN, M.

P 157 O 324 P 029 O 369 P 737 O 283, P 328 P 749 P 081, P 789 O 110 P 699 O 120, P 043 O 089 P 729, P 730 P 920 O 329 P 309, P 310 P 195 P 179, P 804, P 811 P 510, P 556 P 728 P 492 P 020 P 864, P 867 P 568 P 354 P 579 P 131, P 479, P 579 P 234 P 623 P 803, P 807 P 589 P 249 P 130 P 184 P 580 P 602 P 626 P 301 P 419 P 142 O 029, O 159, O 160, O 161, O 318, O 320, O 025, O 026, P 248 P 039 O 236 P 955 P 160 P 577 O 247 O 015, O 163 O 339 O 295 O 100

KARKI, P. KAROUI, H. KARUNAWEERA, N. KASBARI, M. KASSAHUN, A. KASTELLAO, K.G. KATAKURA, K. KATO, H. KATO, K. KATO, K.C. KATZ, S. KAUFMANN, F. KAYE, P.M. KAZIMOTO, T.A. KEAN HOOI, K.A. KEDRA, D. KEESEN, T. KEESEN, T.L. KEESEN, T.S. KEESEN, T.S.L. KEL, A. KELLY, H. KENDJO, E. KENNEDY, L. KENT, A. KENT, A.D. KER, H.G. KESHAVARZ VALIAN, H. KESSLER, A. KETELUT, N. KHAJEIAN, M. KHALIL, E. KHALIL, E.A. KHAMESIPOUR, A. KHANAL, B. KHANDELWAL, K. KHANRA, S. KHARE, P. KHARE,, P. KHATAMI, S. KHATRI, M.L. KHOURI, R. KIAFAR, 5.

KIDAR, A. KIKA, B. KIM, O.M. KIMA, P.E. KINGHORN, A.D.

O 111 P 517 O 217, P 027, P 028, P 668, P 669 O 221, O 124 O 216, O 248 P 631 P 026 P 026 O 311 O 081, O 082 P 350 O 297 O 162 O 340 P 865, P 868, P 872, P 874, P 875, P 878, P 879 O 306 O 205, P 232 P 255 P 286 P 128 O 050, P 273 P 059 O 109 P 457 O 213 O 098 P 151 P 458, P 794 P 129, P 434, P 903 O 211 P 367 P 253 O 195, O 127 P 627 O 356 P 692 O 211, O 080, O 318, P 085, P 299, P 300 O 002, O 015, P 384, P 672 O 171 P 802 O 120, P 043, P 388, P 514 O 089 O 239 O 123 O 291, P 618 P 340 P 905 P 295 P 534 O 005 O 308

KIRSZTEIN, O. KISSINGER, J.C. KITA, K. KITAHARA OLIVEIRA, M.Y. KJAER, V.K. KLATT, S. KOBETS, T. KOBOR, M.S. KOBS, T.L. KOCHHAR, S. KOHIDAI, L. KOHL, K. KÖHLER, K. KOKICI, M. KOLARICH, D. KOLES, N. KOLLI, B.K. KOLOURA, I. KONG, P. KONTHUR, Z. KOPYDLOWSKI, K.M. KOREN, R. KORENAGA, M. KORLACH, J. KOSTALOVA, T. KOTHA, S. KOUROU, K. KOWALEWSKI, M.M. KRAIEM, M. KRAJA, D. KRATOCHVILOVA, T. KRAVAR GARDE, L. KRAWCZAK, F.S. KRAYTER, L. KREISHMAN DEITRICK, M. KREISHMAN DETRICK, M. KRIEGER, H. KRISHNAPPA, H. KROPF, P. KRSTICEVIC, F.J. KRUMBIEGEL, G.B. KUAMAR, D. KUHLMANN, F.M. KUHLS, K. KULSHRESTHA, A. KUMAR, A. KUMAR, D. KUMAR, G.

O 216 O 200 O 179 O 234 P 080 P 019 O 049, O 086, O 260 O 353 P 898 O 109, O 021 P 017 O 035 P 694 P 295, P 611 P 019 O 160 P 449 P 487 P 519 P 019 O 363 O 147 O 038 O 107 P 029 O 308 P 318 P 814 P 501 P 295, P 611 P 717 P 544 P 700 P 671 O 362, O 363 O 289 P 747 P 596 O 237, O 133, P 410, P 437 P 121 P 164 P 582, P 597 O 074 O 102, O 137, O 139, P 694, P 711, P 877 P 364 O 109, O 021 O 026 P 154, P 495 P 284

KUMAR, M.R. KUMAR, N. KUMAR, R. KUMAR, V. KUMAR, V.N. KUREY, I. KUSHAWAHA, P.K. KUSHAWAHA,, P.K. KUTLUBAY, Z. KUYPERS, K. KWAKYE NUAKO, G. LA GIGLIA, M.A. LAAMRANI, A. LABRUNA, M.B. LACERDA, H.G. LACEY, C.N.J. LACHAUD, L. LADEIRA, L.O. LAFORGE, M. LAGE, D. LAGE, D.P. LAGE, P. LAGE, P.S. LAGO, E. LAGO, E.L. LAGO, J.H.G. LAHAV, T. LAHMEDI, R. LAI A FAT, R. LAI A FAT, R.F. LAKHAL NAOUAR, I. LAKSHMI, B.S. LAL, C. LAL, D. LAL, S. LAM, L.L. LAMBERT, M. LAMBERTZ, U. LAMI, P. LAMMEL, E.M. LAMOTHE, J. LANA, M. LANA, R.S. LANDGRAF, V.O. LANG, T. LANGHOR, I. LANGUE, R. LANNA, M.F.

P 450 O 057 O 206, O 207, O 299, O 265, P 086 P 063 P 130 O 049, O 260 P 388 O 089 P 360 P 460 P 946 P 050 O 253 P 700 P 128 P 285 O 162 P 457 P 562 O 330 P 375 P 226, P 520 O 157, P 018, P 123, P 362, P 376 P 375 P 226, P 520 P 018, P 376 O 278, P 024, P 423, P 785 P 134 P 641 O 348 O 138 O 098 P 151 O 026 O 088 O 057 P 667 P 154 O 353 O 303 O 054, P 180 O 221, P 140 P 334 P 221 P 067 P 758, P 764 P 479 O 047, P 430 P 448 P 098 P 197, P 199

LANZA, J.S. LAOUINI, D. LARA SILVA, F.O. LARANJEIRA, D.F. LARANJEIRA, E.T.S. LAROSA, R. LARRAGA, V. LAUKENS, K. LAULETTA LINDOSO, J.A. LAUNAY, D. LAUNOIS, P. LAURANCE, J. LAURENT, M.D. LAURENTI, M.D. LAUTELA, J.A. LAUTENSCHLA, H.R. LAVITSCHKA, C.O. LAWYER, P. LAWYER, P.G. LAZO ESCALANTE, M. LAZO, M. LE CACHEUX, C. LEAL JUNIOR, F.G. LEAL NETO, O.B. LEAL, G.G.A. LEAL, S.M. LEAO, E.F. LEATHERBARROW, R.J. LECOEUR, H. LEE, G.I. LEEDS, S. LEEFLANG, M. LEGENTIL, L. LEITE, A.B. LEITE, A.C.L. LEITE, C.G. LEITE, H.S. LEITE, J.C.S. LEITE, J.H.A.C. LEITE, J.P.V. LEITE, M.D.X. LEITE, M.O. LEITE, R.S. LEITE, S.T.V.C. LELES, D. LEMAIRE, J. LEMESRE, J.

P 492 P 058, P 059, P 922 P 759, P 765 O 181, P 524 P 209, P 211 P 658 O 348, P 148, P 152, P 157 P 385 P 231 P 596, P 636, P 642 O 273 O 092, O 027 P 193, P 422 P 424 P 142, P 146, P 167, P 169, P 291, P 394, P 793, P 855 P 212, P 736, P 771, P 852 P 431 P 793 P 861 O 297 O 161 O 121 P 245 P 535 O 164 O 181 P 664 P 071 P 497, P 510, P 556 P 902 O 152 O 047 P 148 O 114 O 060 O 190 P 528, P 594 P 541, P 564 P 812 P 755 O 183 P 054, P 056, P 686 P 362 P 833 P 740 P 416, P 552 P 810 P 840 P 058 P 159

LEMICHEZ, E. LEMOS, E.M. LEMRANI, M. LEON GUERRERO, D. LEON, L. LEON, L.L. LEOPOLDO, P.T.G. LESCANO, A.G. LESSA, M. LESSA, M.M. LEUNG, D.T. LEUTENEGGER, C. LEVCHENKO, A. LEZAMA DAVILA, C. LIA, R.P. LIAO, L. LIARTE, D.B. LIBURKIN, T. LICIO, C.S.A. LIEBEL, S.M.R.S. LIGEIRO, D. LIMA JUNIOR, M.S.C. LIMA JUNIOR, D. LIMA, A.C. LIMA, A.L. LIMA, A.M. LIMA, A.P.C.A. LIMA, B.D. LIMA, B.S. LIMA, C. LIMA, C.A.A. LIMA, E.F. LIMA, F.P. LIMA, I.S. LIMA, J.G.B. LIMA, J.P.M.S. LIMA, J.R. LIMA, J.T.R. LIMA, J.V.F. LIMA, J.W.O. LIMA, L.D.R. LIMA, L.M. LIMA, L.R. LIMA, L.V. LIMA, M.A. LIMA, M.C. LIMA, M.S. LIMA, M.S.C.S.

P 430 P 349, P 496 O 253, P 015, P 016, P 092 P 417 P 427 P 523 P 250, P 261, P 456 O 351 P 680, P 835 P 133 P 871, P 889, P 891 P 886, P 888, P 890, P 897 P 885, P 894 P 278 P 686, P 852 O 371 O 308 O 010 P 055, P 697 P 099, P 256 O 072 P 509 P 236 O 341 P 801 O 328 P 759, P 765 P 499, P 502, P 506, P 677 P 147 P 263 O 071 P 263 P 222 O 126, P 095 P 850 O 293 P 209 P 842 P 731 O 181, P 524 O 036 P 285 P 278 P 809 P 422, P 424 P 813, P 815, P 892 P 876, P 880 P 169, P 855 P 065 P 571 P 488 P 193, P 422 P 424 O 108, O 057, O 261 P 933 O 043, P 739, P 853 P 775

LIMA, N. LIMA, P.M. LIMA, R.B. LIMA, S.C. LIMA, S.S. LIMA, V.M.F. LIMA, W.P. LIMAM, S. LINDOSO, J.A. LINDOSO, J.A.L. LINO, M.T.B. LIOTTA, D.J. LIPIK, V. LIPOLDOVA, M. LITZKE, L. LIU, D. LLANES ACEVEDO, I.P. LLANES, P. LLANOS CUENTAS, A. LLANOS, A. LLEONART, R. LLULL, J. LO VERDE, V. LOBO, C.H. LOCATELLI, F.M. LOCKWOOD, D.N. LONARDONI, M.V.C. LONGCHAMP, J. LOPES DO CARMO, L.A. LOPES MORI, F.M.R. LOPES, A.P. LOPES, D.M. LOPES, I.D. LOPES, J.D. LOPES, L. LOPES, M.V. LOPES, R. LOPES, R.M. LOPES, U.G. LOPES, X.G.C. LOPEZ GONZALVEZ, A. LOPEZ SANCHEZ, A. LOPEZ, A. LOPEZ, L. LOPEZ, Y. LORDELO, M. LORENZ, B. LORIA CERVERA, E.N.

O 356 P 262 P 664 O 167, P 479 P 519 P 740 P 645 P 394, P 402 P 395 P 736 P 407 P 586 O 227, P 472 P 371 P 241 P 577 P 504 O 045, O 097 P 549 O 049, O 086 O 260 P 694 O 373 P 188 P 009 O 267, P 144, P 245, P 397, P 438, P 535 O 254 P 009 P 075 P 400, P 693 P 622 O 038 O 085 P 233 P 053 P 698, P 712 P 787 P 726 P 141 P 896 P 172, P 175 P 156 P 805 P 559, P 625 O 182, O 336, P 508 O 312 O 037, O 275, P 155, P 183, P 222, P 304 P 896 P 373 P 461 P 464, P 467 O 280 O 361, O 380 O 364 P 025, P 046, P 047 O 238 O 333 P 647

LOSADA BARRAGAN, M. LOURDES, R.A.E. LOUTET, M. LOUZIR, H. LOVE HOMAN, L. LOVELL, A.M. LU, W. LUBBAD, M.Y. LUBY, S.P. LUCAS, C.M. LUCHEIS, S.B. LUGO, D. LUGO, D.A. LUISI JUNIOR, C.M. LUN, Z. LUPO, T. LURA, T. LUTGEN, P. LUVIZOTTO, M.C.R. LUZ, E. LUZ, N. LUZ, N.F. LUZ, S.L.B. LYE, L. LYNEN, L. LYONS, P. LYRA, M.R. M, S.L. M, S.L.L. M. PEREIRA, D.C. MACEDO SILVA, S.T. MACEDO, B.M. MACEDO, D.S. MACEDO, E.C. MACEDO, L.A. MACEDO, S.R.A. MACHADO DE ASSIS, T. MACHADO DE ASSIS, T.S. MACHADO COELHO, G.L. MACHADO COELHO, G.L.L. MACHADO, A.L. MACHADO, A.R.S.R. MACHADO, C.M.T. MACHADO, E.M.M. MACHADO, F.C. MACHADO, G.F. MACHADO, L.F.M. MACHADO, L.M.

O 325 P 035 O 003, P 377 O 236, O 363, P 039, P 958 O 302 P 289 P 110 P 014 O 018, O 062, O 172, P 741, P 796 O 351 P 745 P 298, P 302 P 203 P 247 P 055, P 697 O 187, P 064, P 400, P 693, P 837 O 194 P 469 P 634 P 635 O 096, P 098, P 776 P 136, P 343 O 051 P 920 O 074, O 142, O 351, P 116, P 277 O 262, O 263 O 343 P 237 P 597 P 483 P 863 P 352 P 526 P 888, P 891 P 897 P 885, P 886, P 889, P 890, P 894 P 468, P 770 O 275 P 325 O 264 P 768 P 567 P 214, P 795 P 021 P 201, P 534 P 347 P 533 P 512 P 176, P 229, P 247, P 252 P 032, P 049, P 056, P 196, P 687 P 165 P 191, P 253 P 145

MACHADO, N. MACHADO, P. MACHADO, P.A. MACHADO, P.R. MACHADO, P.R.L. MACHADO, R.Z. MACHELARD, A. MACHINI, F.K. MACIEL REZENDE, C.M. MACIEL, B.L. MACIEL, J. MACKENZIE, B. MACKENZIE, C.D. MACORIS, M.L.G. MACRÌ, D. MADDUGODA, M.P. MADEIRA, I. MADEIRA, M.F. MADHUBALA, R. MAES, I. MAES, L. MAGALHAES, A. MAGALHAES, A.C.S. MAGALHAES, A.S. MAGALHAES, F.B. MAGALHAES, L.K. MAGALHAES, M.A.F. MAGALHAES, R.D.M. MAGALHAES, V. MAGALHAES, V.P. MAGILL, A. MAHAJAN, R. MAHFUZ, F. MAHMOUDI, A.R. MAIA ELKHOURY, A.N. MAIA, C. MAIA, F.C. MAIA, J.M. MAIA, R. MAIA, R.G. MAJI, M. MAJUMDAR, S. MAJUMDER, H.K. MAJUMDER, S. MAKOWIEKI, M.E. MAKURATH, M.A. MALAVIYA, P. MALEKI, 3.

P 476 P 204, P 480, P 527 P 459 O 277, O 050 O 169, P 134 O 166, P 337, P 423, P 785 P 249 O 273 P 282 P 538, P 553 P 255 P 919 O 070 P 448 P 748 O 187 P 430 P 310 O 122, P 824 P 090, P 415 O 088 P 062 O 146, P 451, P 452, P 460, P 636, P 642 P 023 P 939 P 264 P 592 P 271 P 583 P 725 P 018 P 087 P 516 O 289 O 108, O 057, O 261 O 064 O 318 O 229 O 219, P 453, P 887 P 640 P 726 P 577 P 492 O 154 P 584 P 359, P 365 P 584 P 869 O 283 O 012, O 015, O 316, O 357 P 340

MALHEIRO, A. MALHEIROS, A.F. MALIK, M.D. MALONE, J.B. MALTA, H. MALTA, M. MALVIYA, P. MANAFI, H. MANAGIDO, M.B. MANCERO, T. MANDAL, G. MANDAL, S. MANNA, L.L.M. MANNA, M. MANNAER, A. MANNAERT, A. MANS, D. MANS, D.R. MANSINHO, K. MANTECA ACOSTA, M. MANTECA ACOSTA, M. MANTILLA, J.C. MANUEL, C. MANZANO, J.I. MARCELINO, A.P. MARCET, P. MARCILI, A. MARCO, J.D. MARCONDES, C.B. MARCONDES, M. MARCOS, S.F. MARCUS, J. MARGIOTTA, A. MARIA, A.L. MARIALVA, E.F. MARIANTE, R.M. MARIN, D. MARINHO JUNIOR, J.F. MARINHO JUNIOR, J.F. MARINHO, C.C. MARINHO, P. MARINO, E. MARION, E. MARKIKOU OUNI, W. MARKIKOU, W. MARKING, U. MARKUS, R.P. MARLOW, M.A.

P 258 P 809 P 671 P 901 P 343 P 786 O 163 P 701 P 750, P 751 O 380 O 004, P 070, P 107, P 248 P 070, P 107 P 425 P 802 O 344 O 002, O 143, P 672 O 098, P 458, P 794 P 151 P 805 P 806, P 822 O 097, P 778 P 560, P 573 P 122 P 413 P 764 O 252 P 685, P 809 O 038, P 485 O 358 P 054, P 056, P 394, P 395, P 686, P 687, P 736, P 771, P 793, P 852 P 209, P 211 P 277 P 064 P 754, P 755 P 924 P 923, P 925, P 926, P 927 O 301 P 859 P 850 P 906 P 585 P 795 P 278 P 769 O 366 O 208 P 039 O 023 O 048 P 866

MARQUES PAULO, E. MARQUES, C. MARQUES, C.S. MARQUES, L.H.S. MARQUES, M. MARQUES, M.C.S. MARQUES, M.J. MARQUES, P.R. MARQUEZ, E.S. MARQUEZ, R. MARQUIS, H. MARR, A.K. MARRA, A.K. MARTEL, C. MARTELETO, B. MARTELETO, B.H. MARTIN MARTIN, I. MARTIN, D. MARTIN, O. MARTIN, R. MARTIN, V. MARTINEZ FLOREZ, A. MARTINEZ, A. MARTINEZ, A.M.B. MARTINEZ, D. MARTINEZ, D.Y. MARTINEZ, E. MARTINEZ, M.A. MARTINEZ, M.F. MARTINEZ, R. MARTINKOVIC, F. MARTINS FILHO, O. MARTINS FILHO, O.A. MARTINS MELO, F.R. MARTINS, A. MARTINS, A.C. MARTINS, A.C.C. MARTINS, C. MARTINS, D.A. MARTINS, D.R. MARTINS, D.R.A. MARTINS, E.P. MARTINS, J.C.D. MARTINS, L.E. MARTINS, L.F. MARTINS, M.S. MARTINS, N.V.D.N. MARTINS, T.F.C.

P 228, P 241 O 331 O 220, O 319, P 490, P 772, P 786 P 816 P 900 P 453 P 543 P 434, P 533, P 538, P 553, P 816 P 408 P 441, P 444, P 742 P 652, P 653, P 774 O 147, O 258 O 159 O 003, P 377 O 353 O 338 P 292 P 194 O 244, O 245, P 763 P 596 O 193, P 791 O 035 P 494 O 214, P 068 P 075 P 769 P 208 P 144 O 267 P 535 P 578 P 882 O 097, P 778, P 814 P 707 O 135 O 293 O 276, P 069, P 194, P 197, P 199, P 643 O 043, P 739, P 853 P 887 P 037 P 090 P 164 O 182 P 147 P 278 P 285 P 955 P 764 P 520 P 351 O 039, O 100 P 949 P 472

MARTINS, T.M. MARTINS, T.Y. MARTINS, V. MARTINS, V.P. MARTINS, V.T. MARTIS, N. MARTY, P. MARUYAMA, S.R. MARZOCHI, K.B.F. MARZOCHI, M.C.A. MARZOUKI, S. MASATAKA, K. MASCARENHAS, N.M.F. MASHAYEKHI, 4.

MASPERO, R.C. MASSAFERA, R. MASSON, J. MASTRANGELO, A.V. MASUCCI, M. MATHEOUD, D. MATHIAS, F.A. MATHIAS, F.A.S. MATLASHEWSKI, G. MATOS GUEDES, H.L. MATOS, D.C.S. MATOS, M.F.C. MATOS, P.C.M. MATOS, V.T.G. MATRANGOLO, F.S.V. MATSUMOTO, Y. MATTA, V.L. MATTOS, M.S. MAURICIO, I. MAURICIO, I.L. MAZUMDER, S. MAZZONI, C. MC.MASTER, R.W. MCCAFFREY, A.P. MCCALL, L. MCCAROLL, J. MCGAHIE, D. MCKERROW, J. MCKERROW, J.H. MCLEAN, D. MCLIDEN, J.H. MCMAHON PRATT, D. MCMASTER, R.W. MCMASTER, W.R.

P 408 P 462 P 375 P 165 P 226 P 018, P 362, P 376, P 520 P 221 O 014, O 366, P 221, P 430 P 323 O 122, P 824 P 824 O 122 P 958 P 485 P 716 P 340 P 956 P 738 P 336 O 095 P 837 O 324 P 072 O 091 P 067, P 129 O 155, O 348, O 349, P 670 P 208 O 274 P 131 P 251, P 479 P 864 P 579 P 099 P 313 O 090, O 370, O 174, O 176, O 177, O 178, P 313 P 793 P 269 O 106, P 711 O 102 O 154 P 320 P 377 O 073 O 155 P 901 O 214 O 306 O 285 O 268, P 749 P 278 P 180 O 003 O 353

MEDEI, E.H. MEDEIROS, A.C.R. MEDEIROS, J.F. MEDEIROS, M.A. MEDEIROS, Z. MEDEIROS, Z.M. MEDINA, V. MEDLEY, G. MEDRANO, F. MEHEUS, F. MEHTA, R.D. MEIRA, C.S. MEIRELES, J. MEJIA, E. MEJRI, T. MEKONEN, W. MEKURIA, A.H. MELBY, P.C. MELENDEZ, V. MELLO, M.N. MELO JUNIOR, O.A.O. MELO, A.L.A. MELO, F.L. MELO, G.D. MELO, G.M.A. MELO, J.A.T. MELO, L.M. MELO, M. MELO, M.A. MELO, M.B. MELO, M.M.

MELO, M.N. MELO, N. MENDES AGUIAR, C.O. MENDES SOUSA, A.F. MENDES, A.R. MENDES, B.P. MENDES, M.S.T. MENDES, T. MENDES, T.A.O. MENDES, V.G. MENDEZ, G.A. MENDEZ, J. MENDEZ, S. MENDONCA, A.L.P. MENDONCA, D.S.

P 559 P 293, P 296, P 608, P 609, P 614, P 617 P 345 P 005 P 149, P 368, P 369 P 493, P 498 P 836 O 041 O 016 O 067, P 691, P 692 O 171 P 082 P 601, P 631, P 911 O 220, O 319, P 490 P 259 P 517, P 580 P 410 P 610, P 751 P 447 P 518 O 139, P 711 O 311 O 276 P 761 P 493 P 369 P 368 P 687 P 440 P 195 O 185, P 396, P 688 P 629 P 842 P 532 P 516 P 606, P 839 O 276, O 112, O 091, O 294, P 051, P 066, P 072, P 200, P 213, P 225, P 244, P 393, P 416, P 492, P 565, P 673, P 674, P 675, P 734, P 783, P 784, P 819 O 350 P 238 O 317 P 846 P 102 P 095, P 648 P 486 P 181 P 139 P 008 P 518, P 607 O 159 P 358 P 269

MENDONCA, I.L. MENDONCA, L.Z. MENDONCA, M.S. MENDONCA, S.C. MENDONCA, S.C.F. MENDOZA LEON, A. MENESES FILHO, A.J.S. MENESES, C. MENEZES, J. MENEZES, J.A. MENEZES, J.P.B. MENEZES, T. MENNA BARRETO, R. MENTEN, J. MENZ, I. MERELLES, S.L. MESA, C.V. MESQUITA LIMA, R.T. MESQUITA RODRIGUES, C. MESQUITA, J.T. MESTRE, G.L.C. MHADHBI, M. MICHALICK, M. MICHALICK, M.S. MICHALSKY, E.M. MICHEL, G. MICHELS, P.A. MICHIELIN, O. MIECZKOWSKI, P. MIEREZ, M.L. MIGLIAZZO, A. MIGOTTO, A. MIGUEL, D.C. MIKROS, E. MILAGRES, F.A.P. MILANI, B. MILANI, N. MILES, M. MILES, M.A. MILLER, E. MILLER, E.N. MILLS, J.E. MILON, G. MINERVINO, A.H.H. MINHARRO, S. MIRA, M. MIRANDA VERASTEGUI, C. MIRANDA, D.E.O.

O 198 P 197, P 199 P 883 P 150 O 274, O 165, P 005 P 474 O 322 O 159, O 160, O 161, O 320 P 146, P 169 P 731, P 735 P 167 P 204 P 124 O 262, O 269 P 601, P 631, P 928, P 957 O 044, P 526 O 280, P 589 P 370 P 124 P 370 P 389, P 390 O 184 P 586 P 311 P 606, P 839 P 788 P 567, P 759, P 765 P 754 P 430 P 598 O 035 O 030 P 666, P 779 O 187, P 400, P 693, P 837 P 351 O 282, P 477 P 419 P 406 P 572 P 619 O 203 P 342 P 711 O 137, O 265, P 308 O 102 O 216, O 065 P 133 O 152 O 284 O 047 P 809 P 856 P 189 P 535 O 267, P 245 O 128, P 940, P 941

MIRANDA, H.C. MIRANDA, J.C. MIRANDA, N. MIRET, J. MIRHENDI, H. MIRO, G. MIRTHA, M. MIRZAEI, F. MISHRA, A. MISHRA, A.K. MISRA, P. MITRA, G. MITSUKA BREGANO, R. MITSUKA BREGANO, R. MITTMANN, J. MITTRA, B. MIYASHIRO, S.I. MIYAZAKI, R.D. MKADA, I. MKANNEZ, G. MOCHLY ROSEN, D. MODABBER, F. MODOLELL, M. MODOLO, J.R. MOFFI, P. MOHAMATH, R. MOHAMED, H.S. MOHAMMED, R. MOKNI, M. MOLINA, N. MOLINA, R. MOLINARO, C.M. MOLLA, L.M. MONACO, N.Z. MONDAL, D. MONDEGO, J. MONDRAGON, K. MONROY OSTRIA, A.M. MONTALVO, A.M. MONTAﾃ前, N.A. MONTASSIER, H.J. MONTE NETO, R.L. MONTE NETO, R.L.D. MONTE NETO, R.L.D. MONTEIRO, E.L.O. MONTEIRO, E.M. MONTEIRO, G. MONTEIRO, G.G.

P 943 O 250 P 024, P 844 P 178 O 270, P 704, P 707 O 318 O 342, P 769 P 628 P 621 P 137, P 661 P 449 P 381 O 108, O 261 P 826 P 787, P 829 P 012, P 354 O 348 P 091, P 266 P 754, P 755 O 138 P 058, P 059 O 282 O 365 O 133, P 410 P 863 O 259 O 092, O 027 O 343 O 262, O 263 P 916 O 363, P 125 P 822 O 244, O 245, P 157, P 757, P 763, P 766 P 415 P 631 P 823 O 209, O 084, O 018, O 021, O 062, O 172, O 266, P 366, P 741, P 802 O 030 O 011, O 196, O 380, P 904 P 595, P 882 O 254, P 062 O 254 P 190 P 339 O 112, P 565 O 145 P 729, P 730, P 733 P 214 P 758, P 764 P 522 P 147 P 286

MONTEIRO, G.R. MONTEIRO, G.R.G. MONTEIRO, J.C. MONTEIRO, M. MONTEIRO, M.B.A. MONTEZZO, L.C. MONTGOMERY, J.M. MONTI ROCHA, R. MONTILLA MATOS, M.L.A. MONTOYA, A. MONTOYA, E.A. MONTOYA, R. MOON, K. MORA, M.C. MORADIN, N. MORAES, J.L.P. MORAES, M.O. MORAES, P.N. MORAES, R.C.S. MORAES, S.C. MORAES, V.S.S. MORAIS, G.B. MORAIS, L. MORAIS, M.H.F. MORAIS, M.H.G. MORAIS, M.R.P.T. MORAIS, R.C. MORAIS, S.M. MORALES, J. MORANNE, O. MOREA, G. MOREIRA, B.P. MOREIRA, C.P.S. MOREIRA, D. MOREIRA, D.R.M. MOREIRA, D.S. MOREIRA, I.M. MOREIRA, J.S. MOREIRA, M.A. MOREIRA, M.A.B. MOREIRA, M.L. MOREIRA, M.R. MOREIRA, N.D. MOREIRA, O.C. MOREIRA, P.A. MOREIRA, P.A.C. MOREIRA, P.R.R. MOREIRA, R.M.

P 255 P 273 P 285 P 128, P 289 P 845 P 476 P 261 P 448 P 680 P 349 P 342 O 342 P 327 O 380 P 180 O 038 P 485 O 324 P 673, P 674, P 734 P 783, P 784, P 819 O 184 P 807 P 906 P 820 P 808 P 502 P 884 P 533, P 900, P 945, P 954 P 731 P 735 P 875 P 335 P 622 P 910 P 221 P 104 P 216 P 632 O 372 P 541 P 564 P 339 P 664 P 037 P 090, P 164 P 634 P 928 P 194 P 292 P 209 P 067 O 091, P 072, P 129, P 240 P 083 P 880 P 815 P 177, P 190 P 939

MORELLI, K.A. MORENO MATEOS, D. MORENO, D. MORENO, J. MORENO, M. MORENO, M.A. MORGADO, F.N. MORGADO, M.G. MORIZOT, G. MORKOWSKI, S. MORTARA, R.A. MORTAZAVI, H. MOSER, B. MOSORE, M. MOSQUEDA, L.A. MOSSER, D. MOTA, M.D.S.S. MOTA, V.Z. MOTALAB, M.A. MOTAMEDIRAD, M. MOTOIE, G. MOTTRAM, J. MOTTRAM, J.C. MOU, Z. MOURA JUNIOR, I.L. MOURA NETO, J.P. MOURA, A.A.A. MOURA, A.C.A. MOURA, A.C.J. MOURA, A.L. MOURA, A.P.V. MOURA, F.B.P. MOURA, F.K. MOURA, V.A. MOURI, O. MOUTA CONFORT, E. MOUTA CONFORT, E. MOWBRAY, C.E. MOYO, D. MUKHERJEE, A. MUKHERJEE, B. MUKHERJEE, S. MUKHOPADHYAY, A. MUKHOPADHYAY, R. MUKHTAR, M. MUKHTAR, M.M. MULLEN, A. MULLEN, A.B.

O 136 P 529 O 307 O 215, O 016, O 046, O 293, O 068, P 934, P 937, P 938 P 859 P 342 O 375 P 148 P 037 O 237 P 228 O 363, P 457 O 194, O 289 O 030, O 305, O 335, P 192 P 085 O 140, O 141 P 946 P 779 O 240, P 204 P 949 P 459 P 796 O 239 P 082, P 911 P 222 O 033, O 071 O 373 P 851 P 719 P 622 O 128, P 940, P 941 O 186 P 181 P 200 P 813, P 815 P 892 P 880 P 572, P 619 P 321, P 322 P 457 O 122, P 090, P 415 P 824 P 636 O 287 O 340 P 336 P 401 P 401 P 314, P 315 O 004, P 070, P 107, P 248 P 401 O 349, P 610 O 138 P 288 P 356 O 115 P 450

MULLER, I. MÜLLER, K.E. MUNARI, D.P. MUNASINGHE, A. MUNDER, M. MUNIER LEHMANN, H. MUNIZ, A. MUNIZ, A.C. MUNIZ, C.R. MUÑOZ, D.L. MUNRO, P. MUNSTERMANN, L. MURILLO, J.D. MURRAY, P.J. MURTA, M.F. MURTA, S.M.F. MUS, A. MUSA, A. MUSA, A.M. MUSA, B. MUSKUS, C. MUSKUS, C.E. MUTHUSAMY, N. MUXEL, S.M. MYBURGH, E. MYLER, P. MYLER, P.J. MYNX, P. MYRIANTHOPOULOS, V. MYRSELI, T. NABI, S.G. NABIL, B.H.H. NABIPOUR, Y. NABUT, L.B. NADAL, J.M. NÆSS, A. NAGEM, R. NAGEM, R.A.P. NAIFF, M.F. NAKAGHI, A.C.H. NAKAMURA, C.V. NAKAO, R. NAKHASI, H. NAKHASI, H.L. NAMAN, C.B. NAOUAR, I. NARA, T. NARDY, V.

O 133, P 410 O 237, P 437 P 230 P 177 P 398 P 670 O 133, P 410 O 284 O 250 O 278 P 436, P 729, P 730, P 732, P 733 O 280, P 327, P 328, P 469, P 589 P 430 O 008 O 280 O 132 P 256 P 099, P 339, P 442 P 627 P 691 O 343, O 356, P 692 O 343 O 254 O 286, O 347, O 164, P 531, P 589, P 951 O 369 O 048, O 129, P 081, P 465 O 071 O 142, O 054, O 349 O 107, O 348 O 142 P 406 P 843 O 222 O 084, O 376 O 289 P 621 P 441, P 444, P 742 P 075 P 230 O 257 P 018 P 258 P 249 P 625 P 026 O 368 O 026 O 308 O 236, P 039, P 153 O 179 P 187

NARVAEZ, M.A. NARVAEZ, M.L.A. NASCIMENTO SILVA, M. NASCIMENTO SILVA, M.M. NASCIMENTO, A.A. NASCIMENTO, A.A.S. NASCIMENTO, B.W.L. NASCIMENTO, D.V. NASCIMENTO, E.G. NASCIMENTO, E.T. NASCIMENTO, F.C. NASCIMENTO, J.C. NASCIMENTO, L.M.D. NASCIMENTO, M.B. NASCIMENTO, M.D.D.S.B. NASCIMENTO, M.P. NASCIMENTO, M.S. NASCIMENTO, M.S.L. NASCIMENTO, M.T. NASCIMENTO, P.P. NASEREDDIN, A. NASKAR, K. NASSEREDIN, A. NATARAJAN, G. NATARAJAN, G.R. NATEGHI ROSTAMI, M. NAURATH, P.E. NAVARATNE, C. NAVARRO, A.L. NAVARRO, I.T. NAVAS, A. NDAO, M. NDUNGU, J. NEEDHAM, J. NEGRAO, F. NEGRAO, F.J. NEGREIROS FILHO, O. NEGREIROS, A.S. NEGRI, A. NEGROMONTE, G.R.P. NEIRA, L.F. NEITZKE ABREU, H.C. NERLAND, A.H. NETO DA SILVA, M.A.C. NETO, C.P. NETO, F.F. NETO, J.F.R. NETO, L.S.

P 040 P 443 P 380, P 474 O 039 O 100, P 833 O 317 P 239 P 708, P 798 P 226 P 391 P 896 P 289 P 165 P 253 P 788 P 270 P 592 P 436, P 729, P 730, P 732, P 733 P 201, P 534 O 180, O 210, O 328 P 253 O 304 P 147, P 289 O 243 P 684 O 032, O 290 P 088 P 455 O 369, O 026 O 368 O 211 P 930 P 670 P 489 O 096, P 441, P 444, P 713, P 715, P 716, P 742, P 787, P 826, P 829 O 147, P 089 O 256 P 366 O 289 P 283 P 893 P 856 P 468, P 618, P 770 O 307 P 883 P 497 P 236, P 849, P 858, P 862 P 230 P 436, P 729, P 730, P 732, P 733 P 176 P 255, P 286 P 128 P 398

NETO, M.B. NETO, O.P.M. NETO, R.B. NETO, R.D.M. NETTO, M.B. NEVES, B. NEVES, F.T.P. NEVES, L.X. NEVES, P.C.B.C. NEWTON, R. NGUNDAM, F. NICODEMO, A.C. NICOLETE, R. NIETO, J. NIETO, P.D. NIHASHI, N.R. NILGES, M. NIVEYRO, C. NOBUSUE, K. NOGUEDA TORRES, B. NOGUEIRA DE MELO, G.A. NOGUEIRA FREITAS, Y.B. NOGUEIRA, B.M.F. NOGUEIRA, F.C. NOGUEIRA, F.S. NOGUEIRA, P.M. NOGUEIRA, R.F. NOGUEIRA, R.S. NOIRI, E. NONATO, F.R. NORONHA, A. NORONHA, E.F. NORWOOD, J. NORWOOD, J.A. NOTREDAME, C. NOTTON, T. NOVAIS, F. NOVICOV, M. NOVO, S.P.C. NOVOZHILOVA, N.M. NOZAWA, S. NUCATOLO, G. NÜHS, A. NUNES PINHEIRO, D.C. NUNES PINHEIRO, D.C.S. NUNES, C.M. NUNES, D.E.C. NUNES, F.F.C.

P 036, P 198 P 020 O 169 P 492 P 022, P 223 O 372 P 565 P 143 P 512 O 360 O 114 O 168 P 435 P 108, P 325, P 747 O 215, O 068, P 934 P 901 O 179 P 403 P 008 O 084 P 595 P 233 P 865, P 868, P 872, P 874, P 875, P 878, P 879 P 358 P 040 P 928 O 275 P 150 P 551 P 269 O 173, O 174, O 176, O 177, O 178 P 487 O 238 O 296, P 929 O 289 O 362 O 205, P 232 O 075, P 277 O 241, O 087 P 485 P 840 O 075 P 189 P 693 O 149 P 499 P 677 P 502, P 506 P 394 P 540, P 687, P 780 P 918 P 496

NUNES, J. NUNES, J.O. NUNES, J.V.S. NUNES, M. NUNES, T. NUNES, V.L.B. NYLEN, S. NYUGEN, B. OCAMPO, C. OCAMPO, C.B. OCAMPO, R. OCAMPOS, H. OCHOA, R. OCHOA, R.A. OCONNOR, A. ODDONE, R. OGAWA, G.M. OGHUMU, S. OJEDA, J. OKWOR, I. OLIVA CORREIA, C. OLIVA, G. OLIVEIRA FILHO, R.D. OLIVEIRA NASCIMENTO, J.M. OLIVEIRA SILVA, C.V. OLIVEIRA NETO, M.P. OLIVEIRA SILVA, A. OLIVEIRA, A. OLIVEIRA, A.F.J. OLIVEIRA, A.G. OLIVEIRA, A.P. OLIVEIRA, A.P.M. OLIVEIRA, B.B. OLIVEIRA, B.C. OLIVEIRA, B.C.M. OLIVEIRA, C.I. OLIVEIRA, C.V.S. OLIVEIRA, D.F. OLIVEIRA, E. OLIVEIRA, E.B. OLIVEIRA, E.F. OLIVEIRA, E.J. OLIVEIRA, F. OLIVEIRA, F.A. OLIVEIRA, F.C.B. OLIVEIRA, F.M. OLIVEIRA, G.G.S. OLIVEIRA, H.C.

P 285 P 851 P 034 P 453 O 341 P 838 O 206, O 207, O 239, O 299 O 241 P 859 O 008 P 328 P 707 O 286, P 531 O 347, P 589 P 884 O 139, P 877, P 881 P 345 O 339 O 270, P 707 O 373 P 525 O 212, O 292, O 320, P 048 P 678 P 283, P 569 P 576 P 269 O 233, O 167, P 234, P 238, P 274, P 551 O 276 P 493 O 330 P 926, P 927 P 846 P 057, P 823, P 832, P 893 P 010, P 411 P 847 O 185 P 010, P 411 P 402 P 036 P 132, P 844 P 106 P 827 O 264, P 358, P 374 P 601 P 846, P 861 P 903 O 160, O 161, O 320, P 958 P 250 P 261, P 456 P 714 P 780 O 183, P 006, P 077, P 182, P 271, P 522 P 391 P 020 P 655

OLIVEIRA, I.Q. OLIVEIRA, J. OLIVEIRA, J.C. OLIVEIRA, J.S. OLIVEIRA, K.R. OLIVEIRA, L.A.M. OLIVEIRA, L.V. OLIVEIRA, M. OLIVEIRA, M.A. OLIVEIRA, M.A.P. OLIVEIRA, M.M. OLIVEIRA, M.P. OLIVEIRA, M.R.F. OLIVEIRA, N.J.D. OLIVEIRA, P.O. OLIVEIRA, P.S. Oliveira, R.B.d.

OLIVEIRA, R.C. OLIVEIRA, R.M. OLIVEIRA, S.D. OLIVEIRA, S.M.D.V.L. OLIVEIRA, T.M.F.S. OLIVEIRA, T.S. OLIVEIRA, W.W. OLIVIER, M. OLLIER, B. OLOBO, J. OLORTEGUI, C.D.C. OLSEN, A. OMMEN, G. ONO, M.A. OOMMEN, J. OPPERDOES, F. ORDOBAS, M. ORE, M. ORIN, C. ORTA, J. ORTA, J.F. ORTEGA, M.E. ORTIGAO, M.R. ORTIZ, R. ORUE, A. OSADA, Y. OSHIRO, E.T. OSMAN, M. OSMAN, O. OSORIO, E.Y. OSORIO, W.G.

P 522 P 805 P 133, P 264 P 419 P 376 P 807 P 135, P 240, P 242 O 170, P 583 P 343 P 945 P 613 P 135, P 242 O 234, O 167, P 274, P 479 O 304 O 296 P 135 P 579 P 593 P 683 P 067 P 743, P 744 P 507, P 626 P 555 P 579, P 851 P 539 P 249 P 073 P 119 O 272 O 029, O 327, O 329, O 338 O 213 O 356, P 627 P 347 O 172 O 006 P 873 O 352 P 384 P 769 P 835 P 544 P 070 P 248 P 203 P 224 P 578 P 474 O 370 P 202, P 569, P 823, P 832, P 846, P 893 O 162 O 265 P 447 P 698

OSPINA, V.E. OSTOLIN, T.L.V. OSTRITZ, A. OSTYN, B. OSTYN, B.J. OTOMURA, F. OTRANTO, D. OUAISSI, A. OUALHA, R. OUELLETTE, M. OVALLOS, F.G. OVANDO, M.O. OWEN, N. OWENS, K. OYAFUSO, L.K. OZAKI, C.Y. OZBEL, Y. ÖZBEL, Y. OZENSOY TOZ, S. PAAPE, D. PACHECO, A.D. PACHECO, L.V. PACHECO, S.G. PACIENZA LIMA, W. PADMANABHAN, P.K. PADRON NIEVES, M. PADRON, M.D.C. PADUA, R.M. PAES LEME, F.O. PAES, R.A. PAGAN, A.J. PAGLIARI, C. PAGLIONI, D.N. PAGNIEZ, J. PAIN, A.M. PAIXAO, F.R. PAIXAO, L.S. PAL, T.K. PALATINK DE SOUSA, C.B. PALATUCCI, A.T. PALMA, M.S. PALMEIRO, M.R. PAN, L. PANDEY, K. PANIAGO, A.M.M. PANSINI, L.N.N. PANTOJA, K.M.B. PAPADAKI, A.

P 560 P 126 P 699 O 078, O 111, O 113, O 015, O 315, O 055 O 357 P 738 O 010, P 330 O 330, P 519 P 184 O 145, O 150, P 079, P 336 P 861, P 912, P 937, P 957 O 054 O 100 O 075, O 351, P 277 P 577 P 637 O 295, O 321, O 174, P 117 O 135 O 295, O 321, P 117, P 287 O 077, O 152 P 056, P 736 P 522 P 344 P 600 O 101 P 017, P 331, P 338 O 144 P 362 O 186 P 090 O 121 P 639 P 759 P 700 P 159 P 700 P 788 P 020, P 182 P 030 P 470 P 048 P 108 P 037, P 090, P 164 O 308 O 056 O 057, O 061, P 599 P 251, P 579, P 851 P 349 P 810 P 318

PAPADOPOULOU, B. PAPARCONE, R. PAPIEROK, G. PAPIEROK, G.M. PAPPENFUSS, T.L. PAQUET, M. PARANHOS FILHO, A.C. PAREDES, A.O. PARENTE, D.M. PARINANDI, N. PARKINSON, T. PARRADO, R. PARRAS, M.A. PARREIRA, B.L. PARREIRAS, P.M. PASA, S. PASCHOAL, M. PASCOAL XAVIER, M.A. PASCUAL, Y. PASQUALI, A.K.S. PASSERO, L.F.D. PASSOS, J. PASSOS, R.O. PASSOS, S. PASSOS, S.T. PATEL, B.N. PATHAK, N. PATI, H. PATRICK, R. PAUL CHOWDHURY, B. PAUL, C. PAULA, C.D.R. PAULINO, T.P. PAVELY, R. PAVINSKI BITAR, A. PAYS, E. PAZ, G.F. PEACOCK, C.S. PEARSON, R.D. PEDRO, N.F. PEDRO, W.A. PEDROSA, C.M.S. PEDROSA, M.O. PEDROSA, R.B. PEDROSO, R.B. PEINE, K. PEIXOTO, A.A. PEIXOTO, S.A.T.

O 101, O 103, O 025, P 070, P 312 O 212, O 292 P 159 P 068 O 339 O 338 P 808 P 907 O 322 O 308 O 152 P 438 O 097 P 322, P 572, P 619 P 516 O 295, O 321 P 378 P 069 O 276 P 542 P 716, P 826 P 142, P 291 P 641 O 188, P 262 P 576 O 272, O 278, P 204 O 240 P 728 P 602 P 596 O 297 P 584 O 146, P 451 P 095 P 297, P 623 O 193 O 159 P 101 P 700, P 703 O 367 P 110 P 278 P 289, P 896 P 857 P 780 P 848, P 871 P 006 P 892 P 554 O 308 P 818 O 170, P 583

PEIXOTO, V.S. PEÑA, M.S. PENAFORTE, K.M. PENHA, L.L. PENNISI, M.G. PERALES, J. PEREIRA DA FONSECA, I. PEREIRA CARVALHO, R. PEREIRA CHIOCCOLA, V.L. PEREIRA, A. PEREIRA, A.A.S. PEREIRA, B.A.S. PEREIRA, B.S. PEREIRA, C. PEREIRA, C.A. PEREIRA, C.C. PEREIRA, C.W. PEREIRA, D.H.D. PEREIRA, G.C. PEREIRA, G.G. PEREIRA, H.F. PEREIRA, J. PEREIRA, J.C. PEREIRA, L. PEREIRA, L.I.A. PEREIRA, L.O.R. PEREIRA, L.R. PEREIRA, L.S. PEREIRA, M.A. PEREIRA, M.H. PEREIRA, M.M.C. PEREIRA, R.M.S. PEREIRA, T.J.C.F. PEREIRA, V.F. PEREIRA, V.R.A. PERES, A.C. PEREZ VALERO, R. PEREZ VICTORIA, J.M. PEREZ, A.A. PEREZ, J.E. PEREZ, J.G. PEREZ, J.L. PEREZ, M. PERIM, M.C. PERKINS, M.D. PEROSSO, J. PEROTE, M. PERRI, S.H.V.

P 889, P 891 P 848, P 885, P 886, P 888, P 890, P 894, P 897 P 044 P 683 P 263 P 837 P 474 O 331, O 341 P 238 P 601 P 007, P 082, P 505, P 631, P 955 P 658, P 911 O 262 P 812 P 065 P 550 P 786 O 170, P 583 P 194, P 292 P 891 P 888 P 191 P 526 O 030 O 322 O 336, P 471 P 160 O 156 P 038 O 083 P 404 O 167, P 479 O 184 P 223 O 331 P 239 O 317 P 270 P 183 O 198, O 322, P 645 P 539 P 564 P 010, P 541 P 205, P 220, P 265, P 411, P 566 P 480, P 527 P 047 P 025, P 046 P 417 P 778 P 904 P 680 P 379 O 008 P 828 P 366 O 185, P 396, P 688 P 769 P 402

PERSICHETTI, M.F. PERTEGUER, M.J. PERUHYPE MAGALHAES, V. PERVITSKY, D. PESCARINI, J.M. PESCHER, P. PESSANHA, J.E.M. PESSOA, F.A.C. PETERKA, C.R.L. PETERS, N.C. PETERS, S. PETERSEN, A.L.O.A. PETERSEN, C. PETERSON, A. PETITDIDIER, E. PETRI, S.C.S. PETRONI, T.F. PFEILSTICKER, T.R. PIANTEDOSI, D. PIAZZA, F.M. PIAZZA, M. PICADO, A. PICONE, A. PIETRA, R.S. PIMENTA, P.F. PIMENTEL, J.D. PIMENTEL, J.P.D. PIMENTEL, M.I. PIMENTEL, M.I.F. PINHEIRO, C.G.M. PINHEIRO, F.G. PINHEIRO, L. PINHEIRO, L.S. PINHEIRO, M.P.G. PINHEIRO, M.S.B. PINHEIRO, P.G.M. PINHEIRO, S.B. PINHO, F.A. PINHO, M.C.S.O. PINNA, R.A. PINTO, A.J.W. PINTO, B.F. PINTO, E.F. PINTO, E.G. PINTO, I.S. PINTO, J. PINTO, M.C. PINTO, M.D.C.

P 837 P 494 O 276, P 069, P 643 P 518 P 747 O 205, P 232 P 821 P 345, P 920, P 923, P 924, P 925, P 926, P 927 P 681 O 121 O 171 P 106, P 326 P 884 P 094 P 159 P 371 P 745, P 873 P 900 P 048 O 093 P 064, P 400, P 693 O 055 O 092, O 027 P 643 P 103 P 956 P 258 P 150 O 237, P 237 O 165 P 006, P 020, P 182 P 224, P 258, P 260 P 309 P 120 P 915, P 917 P 649 P 006 P 828 P 091 P 266, P 267 O 129 P 210 P 316 P 358 O 156, P 215, P 235 P 409, P 482, P 612 P 818 P 262 P 700, P 703 P 492

PINTO, M.R. PINTO, N.F.S. PINZAN, C. PIPERO, P. PIQUEMAL, D. PIQUET PELLORCE, C. PIRAJA, G.V. PIRANGY, F.R.F. PIRES, F.R. PIRES, M.A. PIRES, S.F. PIRMEZ, C. PIROVANI, C.P. PISHGOOEI, 2.

PISSARRA, H. PIVETTA, O.S. PLEBAN, K. POCHE, D. POCHE, R. POHLMANN, A.R. POIRIER, G. POLAT, E. POLATOGLOU, E. POLETTO, F. POLITOU, A. POLMAN, K. POMARES, C. POMBA, C. POMPEU, M.M.L. PONTE SUCRE, A. PONTES DE CARVALHO, L.C. PONTES, C.S. PONTES, E.R.J.C. PONTES, N. PONTES, N.N. PONTES, S.M. POPOV, V. PORROZZI, R. PORTER, C.H. PORTES, M.G.T. PORTO, A.L.F. PORTUS, M. POSTIGO, M. POTET, J. POZZOLO, E.M. PRADO, A.P. PRADO, A.P.D. PRADO, W.S.

P 557 P 842 P 198 P 611 O 345, P 059 P 122 P 863 P 583 P 538, P 553 P 476 P 066, P 349 P 206 O 234, O 167, P 274 P 099 P 340 P 772 P 957 P 482 P 651 P 651 P 654 P 558, P 559 P 336 P 360 P 101 P 558, P 559 P 318 P 144 O 014, O 366, P 221, P 430 O 319 P 034, P 174 P 331, P 338 O 183, P 006 P 695, P 698, P 712 P 251 P 884 P 255, P 273, P 278, P 286 O 051, P 013 O 053 O 184, O 325 O 252, P 899 P 869 P 640 P 944, P 947 P 157 O 203 P 826 P 748 P 841 P 864, P 867

PRAJAPATI, V.K. PRASASD, N. PRATA FONSECA, A.M.D.R.B. PRATAP, J.V. PRATES, D.B. PRATLONG, F. PRATTI, J.E.S. PRESBER, W. PRESTES, S.R. PREVEL, F. PRICE, H.P. PRINA, E. PROFETA DA LUZ, Z.M. PROTEASA, G. PRUZINOVA, K. PUIG, M. PULIDO, S.A. PULIDO, X. PULLUKCU, H. PUPLAMPU, N. PURCELL, N. PURI, S. QUARESMA, J. QUARESMA, P.F. QUEIROZ, A. QUEIROZ, A.C. QUEIROZ, D.C. QUEIROZ, D.P.S. QUEIROZ, D.S. QUEIROZ, I.T. QUEIROZ, J.W. QUEIROZ, L.H. QUEIROZ, M.F.M. QUEIROZ, P.V. QUEIROZ, P.V.S. QUIJADA, J.D. QUINNELL, R. QUINTAL, A.P. QUINTAL, A.P.N. QUINTANA, M.G. QUINTAO, L.C. QUINTELA, G. QUINTELA, L.P. QUINTELLA, L.P. QUIROGA, M.M. QVIT, N. R SOBRINHO, A.C. RABAN, R.

O 315 O 113, P 364 O 171 P 616 P 449 O 326, P 118, P 844 O 135, O 014, O 221, O 351, P 140, P 272, P 905, P 908 P 162 O 135 P 583 O 052, P 112 O 077 O 284 O 374 O 035 O 248 P 450 O 283, P 328 P 467 P 117, P 287 P 946 P 366 P 596, P 642 P 493 P 225 P 846 O 276 O 240, O 166, P 423, P 785 P 593 P 440, P 488, P 528, P 594 O 317 P 543 P 202 P 521 O 227 P 278 P 147, P 289 P 780 P 820 P 147 P 285 P 257 O 008, O 042, O 197, P 513 P 176 P 161, P 229, P 247, P 512 O 045, O 095, O 097 P 950 P 036 P 090 P 164 P 949 O 282 P 897 P 654

RABELLO, A. RABELLO, A.L.T. RABHI, I. RABHI, S. RABIDAS, V.N. RACHID, M.A. RACINE, G. RAFATI, S. RAHMAN, K.M. RAHMAN, M. RAHMAN, M.M. RAHMAN, M.W. RAHMAN, R. RAI, A. RAI, K. RAI, M. RAI, R. RAIZER, J. RAMALHO ORTIGAO, J.M. RAMALHO ORTIGAO, M. RAMANATHAN, V. RAMASAMY, G. RAMASAWMY, R. RAMDAS, S. RAMESH, V. RAMILO, D. RAMIREZ, L.E. RAMOS JUNIOR, A.N. RAMOS SANCHES, E.M. RAMOS SANCHEZ, E.M. RAMOS, A.P. RAMOS, A.S. RAMOS, C. RAMOS, F. RAMOS, G.S. RAMOS, M. RAMOS, P.K. RAMOS, P.K.S. RAMOS, R. RAMOS, S.R. RAMOS, W.R. RANASINGHE, P.S. RANASINGHE, S. RANGEL, E.F. RANGEL, M. RANGEL, M.U. RANGEL, O. RANSOM, J.

O 264, P 069, P 228, P 358, P 374, P 768, P 903 O 224 P 587, P 588 O 345, P 059 O 345 O 056 P 051 P 079 O 239, O 118, O 025, P 109 O 018, O 062, O 172, P 741, P 796 P 796 O 222, O 376 P 796 O 109, P 366 P 514 O 111, P 386 O 206, O 207, O 113, O 315, O 063, P 154, P 478, P 483, P 495 O 279 P 893 O 246 O 192 P 596 O 107, O 142, O 054, O 348, O 349 P 189, P 264, P 307 O 098 O 024 P 364 O 220 P 781 P 803, P 807 O 043, P 739, P 853 P 637 P 002, P 091 P 535 P 445 P 887 O 038, P 485 O 311 P 825 P 422 P 193 P 424 P 169 P 854 P 755 P 920 P 670 P 668 O 349 O 155 O 378, P 097, P 696, P 699, P 705, P 725 P 250 P 740 P 489 O 363

RANUCCI, T.R. RAO, M. RAOUANE, M. RASO, P. RASSI, Y. RASTOGI, R. RATNAM, J. RAUHUT, F. RAVEL, C. RAVINDRAN, R. RAVINETTO, R. RAYCHAUDHURI, S. RAZURI, H.R. RE, M.I. REA, M.J. REA, S. READY, P.D. REAL, F. REALE, S. REBBESTAD, K. REBELO, J.M.M. REED, S. REED, S.G. REED, S.J. REGINA SILVA, S. REGINALDO DE SOUSA, M.L. REGINALDO SOUSA, M.L. REGINALDO, M.L. REGIS, W.B. REGIS, W.C. REGIS, W.C.B. REGNAULT, B. REGO, F.D. REIFUR, L. REIMAO, J.Q. REINER, N. REINER, N.E. REINHOLD CASTRO, K.R. REIS CUNHA, J. REIS, A. REIS, A.B. REIS, C.R. REIS, I.A. REIS, L.C. REIS, L.E.S. REIS, P.G.

P 858, P 862 P 596 P 790 P 629 O 318 P 314 O 306 P 694 O 221, O 124, O 255, P 140, P 272, P 457 O 154 O 262 O 194, O 289 P 680 P 208, P 407 P 357, P 666, P 779 P 050 O 191 O 030, O 305, P 155 O 187, P 400 P 230 P 783, P 784, P 819 P 673, P 674, P 675, P 734 O 161 P 576, P 615 O 209, O 090, O 092, O 093, O 293, O 027, O 177, P 313, P 610 O 266, P 250 O 178 O 092, O 027 P 214, P 567 P 878, P 879 P 874 P 865, P 868, P 872, P 875 P 362 P 226 P 376 P 123 P 018 P 520 O 345, P 059 P 225, P 800 P 098 P 505 P 370 P 390, P 418, P 419, P 477 O 054 P 180 P 233 P 236, P 849, P 858, P 862 P 486 O 293 O 091, P 067, P 071, P 072, P 129, P 135, P 143, P 197, P 199, P 240, P 242, P 434, P 496, P 522, P 533, P 714, P 816, P 903 O 103 P 700, P 703, P 900 P 002 P 129, P 434, P 533, P 816 O 081 O 112, O 145, P 565

REIS, V.W. REITHINGER, R. REMPEL, S.S. RENARD, P. RENTERIA, J.A.C. RENTSCH, D. REQUENA, J.M. RESENDE, D.M. RESENDE, L.A. RESENDE, M. RESENDE, M.O. RESSNER, R. RESTREPO, A.M. RESTREPO, C.M. REZENDE, A.M. REZENDE, M.T. REZENDE, S.A. RHAJAOUI, M. RIBEIRO SAMPAIO, R.N. RIBEIRO ALVES, M. RIBEIRO DE SENNA, M.C. RIBEIRO DIAS, F. RIBEIRO GUIMARAES, M.L. RIBEIRO ROMAO, R.P. RIBEIRO, A.J. RIBEIRO, J.B.P. RIBEIRO, J.G. RIBEIRO, J.M. RIBEIRO, K.A.N. RIBEIRO, L.C. RIBEIRO, L.M. RIBEIRO, M. RIBEIRO, M.L. RIBEIRO, M.S. RIBEIRO, P.F.M. Ribeiro, R.A.N. RIBEIRO, R.R. RIBEIRO, S.J.L. RIBEIRO, T.G. RIBEIRO, T.T. RIBEIRO, V.M. RIBEIRO, Y.J.S. RIBOLLA, P.E.M. RICE, R. RICO, C.I. RICO, E. RIDOFF, M. RIERA, C.

P 567, P 795 P 513 P 408, P 563 P 058 P 918 O 072, O 131 P 062 P 135, P 242 P 434 P 197, P 199 O 372 P 293 O 160 O 280, O 365, P 327, P 328, P 469, P 589 P 009 P 135 P 242 P 067, P 072 P 509 O 082, P 126, P 197 P 092 O 083, O 126, P 095, P 105, P 428, P 439, P 530, P 546, P 648 O 184 P 069 P 404 P 083 P 215, P 235 P 261 P 726 P 476 P 095, P 439 P 108 P 029 P 831 P 158 P 823, P 832, P 846 P 441, P 444, P 742 P 550 P 431 P 838 P 683 P 606, P 839 P 433 P 226, P 376 P 018, P 520 P 874, P 879 P 194 P 630, P 931, P 932 P 292 P 436, P 729, P 730, P 732, P 733 O 354, P 057, P 173 O 289 P 573 P 525 O 193 P 084, P 722, P 947

RIGG, C. RIJAL, S. RIJAL, S.R. RILEY, L. RINCON, M.G. RIOS VELASQUEZ, C.M. RIOS, L.A. RIOS, R. RITMEIJER, K. RIVAS, L. RIVAS, M. RIYAD, M. RIZK, Y.S. RIZZO, N.R. ROA, L. ROATT, B.M. ROBERT GANGNEUX, F. ROBERT, P. ROBERTS, S. ROBERTS, S.C. ROBLEDO, M.B. ROBLEDO, S.M. ROBLES, J.J.T. ROCHA DOS SANTOS, C.F. ROCHA NETO, G.B. ROCHA, A.A.A. ROCHA, A.G. ROCHA, A.M.S. ROCHA, F.A. ROCHA, I.C.M. ROCHA, K.F. ROCHA, L.C. ROCHA, M. ROCHA, M.A.W. ROCHA, M.C. ROCHA, M.N. ROCHA, M.O.C. ROCHA, O. ROCHA, R.L. ROCHA, R.R. ROCHA, V. ROCHAEL, N.C. RODRIGUES NETO, J.F. RODRIGUES SILVA, R. RODRIGUES, A. RODRIGUES, A.M. RODRIGUES, A.M.S.

P 895 O 002, O 140, O 143, O 078, O 285, O 015, O 344, O 357, O 067, O 269, P 053, P 382, P 383, P 384, P 386, P 672, P 679 O 111 O 166 P 443 P 923, P 924 P 328 O 258 O 023, O 226, O 262 O 281, P 373, P 461, P 464, P 467, P 475 P 542 O 253 P 202, P 521, P 543 P 605 P 919 P 067, P 071, P 072, P 129, P 143, P 197, P 199, P 239, P 434, P 533 O 303, O 190, P 122 P 207 O 130 O 132 O 365 O 280, O 283, O 286, O 361, O 365, P 327, P 328, P 469, P 589 O 364 P 918 P 933 P 933 P 493 P 854 P 567, P 731, P 795 P 256 P 900 P 787 P 260 P 472 P 296 P 577 P 637 P 632 P 209, P 211 P 310 O 017, P 279, P 280, P 281 P 496 P 372, P 487 O 304 P 255 O 276 O 331, O 341 P 209, P 211 P 428

RODRIGUES, A.P.R. RODRIGUES, B.S. RODRIGUES, C.M. RODRIGUES, D.B. RODRIGUES, E.H.G. RODRIGUES, F.M.D. RODRIGUES, G.M. RODRIGUES, I.A. RODRIGUES, J.C.F. RODRIGUES, J.S. RODRIGUES, K.M.P. RODRIGUES, L.M. RODRIGUES, L.O. RODRIGUES, M.A. RODRIGUES, M.M. RODRIGUES, M.S. RODRIGUES, O. RODRIGUES, P.O. RODRIGUES, S.A. RODRIGUES, V. RODRIGUES, V.T. RODRIGUES, W.B. RODRIGUEZ VEGA, C.A. RODRIGUEZ CORTES, A. RODRIGUEZ, C. RODRIGUEZ, J. RODRIGUEZ, N.E. RODRIGUEZ, N.M. RODRIGUEZ, O.L. RODRIQUEZ, O.L. ROGERS, M. ROGERS, M.B. ROGERS, M.E. ROHE, M. ROHOUŠOVA, I. ROHOUSOVA, I. ROHRLICH, P.S. ROJAS, A. ROJAS, E.M. ROJAS, L.J. ROLAO, N. ROMA, E.H. ROMAN ESCUTIA, M. ROMANHA, A.J. ROMANO, A. ROMAO, A.B.D. ROMAO, R.R.P.R. ROMERO, G.A.S.

P 622 P 206 O 157 P 243 P 623 P 297 P 293, P 296, P 608, P 609, P 614, P 617 P 212 P 729, P 730 P 445 O 310, P 103, P 352, P 399, P 414, P 432 P 755 O 167 P 141 P 876, P 880 O 182, P 185, P 508 P 633 P 898 O 331 P 202 P 612 P 297 O 330 P 715 P 243 P 646 P 075 P 578 P 573 P 013 P 624, P 914 P 127, P 203, P 254, P 257, P 578 P 302 P 298 O 332, O 042, P 791 O 298 O 193 P 019 O 245 O 244, P 717 P 029 O 118 P 836 P 727 O 147 P 453 O 121 P 113 P 515 O 297 P 263 O 156 O 359, O 296, P 921, P 929

ROMERO, I. RONET, C. ROOS, D. ROOS, D.S. ROSA CRUZ, C.F. ROSA CRUZ, M.F. ROSA, D.S. ROSA, F.A. ROSA, J.R. ROSA, M.D.S.S. ROSA, M.E. ROSALES, M. ROSANDISK, M.L. ROSAS, J. ROSELINO, A.M. ROSELINO, A.M.F. ROSENTHAL, E. ROSSI BERGMANN, B. ROSSI, C.N. ROSSI, M. ROSTAN, O. ROUAULT HARDOIN, E. ROUAULT, E. ROWTON, E. ROY, G. ROY, K. ROY, L. ROY, S. ROYO, M. ROZZA, D.B. RUBIANO PEREA, L.C. RUBIANO, L.C. RUBINO, V. RUFFOLO, B.B. RUGANI, J.M.N. RUGGIERO, G. RUI LUVIZOTTO, M.C. RUIZ SANTAQUITERIA, M. RUIZ, D. RUIZ, J.C. RUTHEL, G. SA, R.G. SA, V.M.H. SA, W.T. SAADI, Y. SABAJO, L. SABAJO, L.O. SABATE, D.

P 515 O 035, O 074, O 273, O 351, P 112, P 115 O 142 O 087 P 652, P 653, P 774 P 774 P 652, P 653, P 774 P 193 P 054, P 056, P 686 O 097 P 445 P 134, P 337 O 099, P 737, P 859 P 150 P 542 P 201, P 534 P 546, P 648 P 221 O 116, P 208, P 555, P 558, P 600 P 394 O 035, O 052, P 112 O 303, P 122 O 284 O 047 O 161 P 336 P 030 O 067 O 235, P 030, P 088, P 382, P 401, P 802 P 467 P 177, P 268 P 737 O 099, P 859 P 048 P 829 P 225 P 048 P 448, P 928 O 307 P 132 P 135 P 242, P 256 O 087 P 067 P 572, P 619 P 885, P 894 P 889, P 891 P 888 P 501 O 098 P 151 O 110

SABERI, S. SABINO, K.C. SABINO, K.C.C. SABIO, P.B. SABOIA VAHIA, L. SABROZA, P.C. SACKS, D. SACKS, D.L. SACRAMENTO, L. SADLOVA, J. SAENZ, E. SAFI, N. SAFI, N.U. SAGICA, F.D.E.S. SAHA, P. SAHAL, D. SAHASRABUDDHE, A.A. SAHEKI, M.N. SAID, R.F. SAID, R.F.D.C. SAKRU, N. SAKURAI, T. SALA, T.N. SALAH, A.B. SALAM, M.A. SALAUN, P. SALAZAR, A.M. SALAZAR, L.M. SALAZAR, V.A. SALCEDO, J.M.V. SALDAÑA, A. SALDANHA, A.C. SALDARRIAGA, O.A. SALES, A.D. SALES, K.G.S. SALES, R.H.F. SALGADO, A. SALGADO, K. SALGADO, V. SALIBA, J.W. SALIBA, S.W. SALIH, M.A. SALJOUGHIAN, N. SALMON, D. SALOMON, M.C. SALOMON, O. SALOMON, O.D.

P 301, P 913 P 307 P 408 O 007 O 325, P 243 P 124 P 743, P 744, P 756, P 954 O 206, O 207, O 299, O 297 O 075, O 121 O 210 O 249 O 251 P 835 O 379 P 690 P 949 P 802 P 466 P 042 O 104, P 130, P 284, P 602 P 090 P 164 P 788 P 821 P 272 P 026 O 129, P 465 O 236, O 236, O 362, O 363, O 125, O 289, P 039, P 125, P 153, P 305, P 306, P 916, P 922, P 942 O 266 P 387 P 446 O 164 P 475 P 747 P 146, P 895 P 132 P 447 P 622 O 128 P 633 O 375, P 797 P 087, P 423, P 785 P 187 P 358, P 374 P 374 O 343 O 025 P 101 P 104 P 825 O 045, O 095, O 097, P 121, P 778, P 792, P 806, P 814, P 822 P 773

SALOTRA, P. SALTO, E. SAMANT, M. SAMJHAWAN, A. SAMPAIO FILHO, A.P. SAMPAIO, E.P. SAMPAIO, R.N.R. SAMPAIO, S.M.P. SAMSON, M. SAN MARTIN, J.V. SANCHES, F. SANCHES, L.C. SANCHEZ TEJEDA, G. SANCHEZ GOMEZ, M. SANCHEZ MURCIA, P. SANCHEZ MURCIA, P.A. SANCHEZ, A.P.C. SANCHEZ, C. SANCHEZ, J.F. SANCHEZ, M.A. SANCHEZ, M.C.A. SANCHEZ, M.V. SANDERS, M. SANJOBA, C. SANTA BARBARA, M.C.A. SANTABARBARA, M.C.A. SANTAMARIA, C. SANTANA, A.L. SANTANA, A.M. SANTANA, A.P. SANTANA, G. SANTANNA, C.M.R. SANTAREM, A.A. SANTAREM, N. SANTIAGO, A.S. SANTIAGO, M.E.B. SANTIAGO, R.C. SANTINI, M.S. SANTINI, S. SANTORO, M.M. SANTOS GOMES, G. SANTOS GOMES, G.M. SANTOS MATEUS, D. SANTOS OLIVEIRA, J.R. SANTOS, A.M.A. SANTOS, B.A. SANTOS, B.N. SANTOS, C.

O 113, O 022, O 171, O 026, P 363, P 364, P 671 O 016 O 101 P 794 P 677 O 234 P 570 P 748 O 303, P 122 O 016 P 291 P 396, P 688 P 882 O 325 P 525, P 529 O 307 P 918 O 215, O 046 P 835 P 203 O 375, P 127, P 254, P 257, P 259, P 342, P 578 P 341 P 104 P 053 O 090, O 370, O 174, O 177, O 178 P 579 P 579 O 106 P 481 P 481 P 740 P 187 P 528 P 095 P 079 O 293 P 099 P 398 P 548 O 045, O 095, O 097, P 778, P 806 P 792, P 814 P 825 P 376 O 336, O 341, P 772 O 182, O 331, P 185, P 508 O 182, O 331, O 341, P 185, P 508 P 228 P 241 P 498 P 858, P 862 P 073, P 119 P 110

SANTOS, C.N. SANTOS, C.R.O. SANTOS, C.S. SANTOS, D.A. SANTOS, D.M. SANTOS, E.L. SANTOS, F.F.M. SANTOS, F.K.M. SANTOS, F.N. SANTOS, G.J. SANTOS, G.J.L. SANTOS, H.D. SANTOS, I.A. SANTOS, I.K.F.M. SANTOS, I.O. SANTOS, J.L.B. SANTOS, J.L.C. SANTOS, L.C.S. SANTOS, L.G. SANTOS, M.B.L. SANTOS, M.F. SANTOS, M.F.C. SANTOS, M.L.B. SANTOS, M.R.A. SANTOS, M.S. SANTOS, O.D. SANTOS, P.A. SANTOS, P.L. SANTOS, P.R.B. SANTOS, P.V.W.G. SANTOS, R.C. SANTOS, R.S. SANTOS, R.S.S.S. SANTOS, T.A. SANTOS, T.A.R. SANTOS, T.C. SANTOS, T.R. SANTOS, Y.C.V. SARAH, H. SARAIVA CORREA, D. SARAIVA, E.M. SARAIVA, L. SARAVIA, N. SARAVIA, N.G. SARDAR, A.H. SARDINHA, F. SARNOSKI JUNIOR, S. SASSI, A.

P 632 O 168, P 435 O 238 P 645 O 367 P 036 P 504 P 433, P 446 P 619 P 775 P 815 P 415, P 712 P 499, P 677 P 502, P 506 P 856 P 655 P 323 P 409 P 813, P 892 P 532 P 526 P 639 P 952 O 319, P 490 P 057, P 893 O 151, P 250, P 456, P 576 P 945 P 440, P 488, P 528, P 594 P 145 P 939 P 549 P 767 P 456 P 953 P 201, P 534 P 795 P 867 P 834 P 817 O 275 P 541 P 564 P 077, P 207 P 634 P 810 P 451 O 146 P 461, P 464 O 301, O 304, P 183, P 246, P 378, P 420 P 812 O 029, O 351, O 258 O 147, P 089 O 256 P 189 P 098 P 184

SATOSKAR, A. SATOSKAR, A.R. SATOW, M.M. SAVAGE, P. SAVAGE, P.B. SAWALHA, K. SAWALHA, S. SCALONE, A. SCHÄFER, C. SCHALLIG, H. SCHALLIG, H.D. SCHALLIG, H.D.F.H. SCHECHTMAN, D. SCHEIRER, J. SCHER, R. SCHESKE, L. Schettini, D. SCHLEICHER, U. SCHLISSELBERG, D. SCHNORR, D. SCHNUR, L. SCHNUR, L.F. SCHOENIAN, G. SCHONIAN, G. SCHÖNIAN, G. SCHOONE, G. SCHOONE, G.J. SCHOUNTZ, T. SCHRAMM, V.L. SCHRIEFER, A. SCHROOTEN, W. SCHUBACH, A. SCHUBACH, A.O. SCHWANKE, K. SCHWEDERSKY, R.P. SCHWEIGERT, A. SCHWEYNOCH, C. SCIO, E. SCODELLER, E.A. SCOFIELD, A. SCOFIELD, K.M. SCOPEL, K.K.G. SCOPELLITI, D. SCORZA, J.V. SCOTT ALGARA, D. SCOTT, P. SCOULICA, E. SEBLOVA, V.H.

O 026 O 308, O 339, O 368, O 369, O 171, P 094 P 577 P 578 P 259, P 342 P 455 P 684 O 232, O 292, P 604, P 947 O 149 O 098, O 258, O 293, P 332, P 458, P 726, P 794 P 151 P 777 O 282 O 289 P 481, P 504 O 258 P 683 O 105, O 064, O 132 O 072 O 278 O 290 P 671 O 137, O 139, O 140, O 141, O 290, P 711 P 364 O 102, O 105, O 135, O 255, P 026, P 092, P 671, P 694, P 724, P 877 O 258, P 332, P 361 P 586 P 647 P 470 O 351, O 166, O 169, P 087, P 337, P 423, P 785 O 267 P 237 O 237, P 037, P 824 O 274, O 122, P 090, P 150 P 422 P 208 P 032, P 049, P 196 O 139, P 711 P 643 P 104 P 864, P 867 P 788 P 568 P 064 P 727 P 298 O 240, O 241, O 272, O 087, P 023, P 204 O 312, O 336 O 248

SEBY, F. SEDEﾃ前, M. SEIFERT, K. SEKAR, A. SEKI, M. SELVAPANDIYAN, A. SEMIAO SANTOS, S.J. SENA FILHO, J.G. SENA, C.A.P. SENA, J. SENA, J.M. SENE, I.S. SENEVIRATNE, K. SERAFIN, L.C. SERAKIDES, R. SERAKIDES, R.S. SERGENT, O. SERRA E MEIRA, P.C.L. SERRA, Q.D. SERRA, T. SERRANO, M.L. SERUFO, A.V. SERVIS, C. SESANA, A.M. SEVA, A. SEVA, A.P. SEVERO, M.M.S. SEVILLA, S. SEYED, N. SGHAIER, R.M. SHAHBAZI, F. SHAIK, J. SHAMS, A. SHAMSIMEYMANDI, M. SHAMSIMEYMANDI, S. SHANKAR, R. SHAPIRO, J.T. SHAREGHI, N. SHARIFI, I. SHARMA, B. SHARMA, M. SHARMA, S. SHARP, A. SHARPE, A.H. SHAW, C. SHAW, C.D. SHAW, J.J. SHAW, M.M.L.

O 080 P 084, P 722 O 079 O 107, O 348 O 370 O 026 P 777 P 842 P 481 P 948 O 114 P 831, P 933 P 052 O 198, P 275, P 290 P 670 P 456 O 276 P 516 O 190 P 812 P 823 P 947 P 494 P 165 P 145, P 191, P 253 O 035 P 213, P 562 O 046 P 937, P 938 P 870 P 769 O 118 P 058, P 059 O 195, O 127 O 074 P 621 P 701 P 701 O 015 P 801 O 318 O 118, P 591 O 356 O 004, P 107, P 248 O 113 P 450 P 094 P 356 P 450 P 053 P 949 O 134, O 242, O 323, O 066, P 658, P 746, P 789 O 323

SHAWKI, A. SHCUBACH, A.O. SHELTON, A. SHIFERAW, W. SHIMABUKURO, P.H.F. SHIO, M.T. SHIPMAN, C.S. SHIRZADI, M.R. SHKURTI, K. SHORTE, S. SHUKLA, S. SILBER, A.M. SILPAPOJAKUL, K. SILVA BARCELLOS, N.M. SILVA JUNIOR, C.F. SILVA SANTOS, L.A. SILVA ARAUJO, M.S. SILVA CARDOSO, L. SILVA COSTA, D.C. SILVA LOPEZ, A. SILVA PEREIRA, R.A. SILVA SANTOS, G. SILVA, A.A. SILVA, A.C. SILVA, A.F. SILVA, A.G. SILVA, A.L.A. SILVA, A.M. SILVA, A.P. SILVA, A.Q. SILVA, A.R. SILVA, A.R.S. SILVA, C.I.M. SILVA, C.J. SILVA, C.L. SILVA, C.P.C. SILVA, C.V. SILVA, D. SILVA, D.A. SILVA, D.C.M. SILVA, D.N. SILVA, E.B. SILVA, E.D. SILVA, E.L. SILVA, E.S. SILVA, E.V.L.P. SILVA, F. SILVA, F.F.

O 070 P 164 O 308 O 216 P 760, P 762, P 800 O 329 O 100 O 195, O 127 P 295, P 611 O 284 O 063 P 473 P 662 O 081 P 080, P 718, P 747, P 847 P 902 O 276 P 166 P 090, P 164 P 338 P 099 O 291 P 898 P 411, P 541, P 564, P 566 P 090 P 569 P 139 P 192, P 626 P 155 P 664 O 169 P 212 P 852 P 547 P 296, P 609, P 614 P 898 P 761 P 176 P 786 P 440 P 626 P 846 P 564 P 149 P 498 P 781 P 683, P 798 P 209, P 211 P 176 P 346

SILVA, F.J. SILVA, F.L.D. SILVA, F.O. SILVA, F.O.L. SILVA, F.O.L.E. SILVA, F.S. SILVA, G. SILVA, G.A.F. SILVA, G.M.C. SILVA, H.C.A. SILVA, I.D.R. SILVA, I.S. SILVA, J. SILVA, J.A. SILVA, J.C. SILVA, J.C.F. SILVA, J.E.S. SILVA, J.F. SILVA, J.H.T. SILVA, J.M. SILVA, J.P. SILVA, J.R.A. SILVA, J.S. SILVA, J.S.F. SILVA, K.B. SILVA, K.G. SILVA, K.L.O. SILVA, K.M. SILVA, K.M.S. SILVA, K.R. SILVA, L. SILVA, L.A. SILVA, L.A.C. SILVA, L.C. SILVA, L.C.S. SILVA, L.F. SILVA, L.O.R. SILVA, L.S. SILVA, L.T. SILVA, M.A. SILVA, M.A.L. SILVA, M.B.A. SILVA, M.D.C.F. SILVA, M.E. SILVA, M.E.B. SILVA, M.G. SILVA, M.L.F. SILVA, M.L.R.

O 128, P 940, P 941 P 949 P 323 O 069, P 052, P 275 P 758 P 788 P 550 P 825 P 569 O 037 P 678 P 424 P 799 P 785 P 945 P 290 P 795 P 758 P 764 P 049 P 032, P 196 P 687 P 650 P 915 P 060 P 720 P 445 O 210, O 328 O 181, P 077, P 524 P 530 P 828 P 940 P 650, P 941 P 396 O 185 P 468, P 770 P 764 P 468, P 770 P 618, P 721 P 311 P 303 P 210 P 907 P 907 P 867 P 865, P 868, P 872, P 874, P 875, P 878, P 879 P 815 P 192 P 091 P 335 P 759 P 788 P 149, P 493 P 368, P 369 P 581 P 581, P 860 P 885 P 888, P 889, P 891 P 886 P 637 P 297, P 623 P 939

SILVA, M.M.M. SILVA, M.R.B. SILVA, M.S. SILVA, M.V. SILVA, O. SILVA, P.H.S.E. SILVA, R. SILVA, R.A. SILVA, R.M. SILVA, R.P. SILVA, R.P.E. SILVA, R.T. SILVA, S. SILVA, S.B.A. SILVA, S.O. SILVA, T.C. SILVA, T.L.V.E. SILVA, T.M.S. SILVA, T.O. SILVA, T.R.R. SILVA, V.C. SILVA, V.L. SILVA, V.M. SILVA, V.P.M. SILVEIRA LEMOS, D. SILVEIRA LEMOS, D. SILVEIRA, A.M. SILVEIRA, F.T. SILVEIRA, H.M.C. SILVEIRA, T.G.V. SILVEIRA, T.L. SILVEIRA, V.M. SILVESTRE, R. SIMA, M. SIMBAIA, M.G.S. SIMOES DIAS, B.R. SINGH, A. SINGH, A.K. SINGH, B. SINGH, D. SINGH, D.P. SINGH, K.P. SINGH, N. SINGH, N.S. SINGH, O.P. SINGH, R.

P 915 P 917 P 613 O 170, P 583 P 297 P 803, P 807 P 670 P 810 O 070 P 678 P 522 P 489, P 793 P 335 P 585 P 512 P 423 P 309, P 317 P 767 O 294, P 200, P 244, P 673, P 674, P 675, P 734, P 783, P 784, P 819 O 139, P 711 P 625 P 222 P 209 P 639 P 594, P 640 P 593 P 489 P 762 P 195, P 561 P 415, P 712 P 921 P 285 P 917 P 138 P 197 P 546 P 864, P 867 P 142, P 146, P 167, P 169, P 193, P 212, P 291, P 422, P 424, P 855, P 902 P 583 P 554 P 268 P 149 O 330, O 372, O 293, P 079, P 726 O 049 P 029 P 918 O 036, P 074, P 326 O 206 O 315, O 058, O 063 O 315 O 015, P 011, P 478, P 749 P 154 O 061 P 667 P 603 O 189 O 206, O 207, O 299, O 268, P 154, P 749 P 667 O 207, O 015, O 265, O 268, P 011, P 086 P 749 P 364 O 078, O 111

SINGH, R.P. SINGH, S. SINGH, S.P. SINGH, T. SINHA, P.K. SIPAHI, O.R. SIQUEIRA FORMIGA, C.C. SIQUEIRA, L.L.D.N. SIQUEIRA, L.L.N. SIREKBASAN, S. SIRIMANNA, G. SIRIWARDANA, Y. SIRIYASATIEN, P. SISSAY, Y. SIVINI, M.A. SJURSEN, H. SKALTSOUNIS, A. SLAPNICKOVA, M. SLEIGH, A.C. SLIM, E. SMIRLIS, D. SMITH, B.A. SMITH, D.F. SMITH, P.L. SNOUSSI, M.A. SOARES, B.R. SOARES, C.R. SOARES, D.C. SOARES, D.F.M. SOARES, F.V. SOARES, G. SOARES, H.S. SOARES, K.A. SOARES, M.B.P. SOARES, M.I.S. SOARES, M.M.C.N. SOARES, M.R.A. SOARES, R. SOARES, R.M. SOARES, R.O.A. SOARES, R.P. SOARES, R.P.P. SOARES, S.C. SOARES, S.R.C. SOBRAL, A.A. SOBRINHO, A.C.R. SOBRINHO, L.S.V. SOBRINO, A.C.R.

O 012, O 015, O 316, O 357, O 055, O 163, P 749 O 093 O 055 P 449 P 186 O 057 P 117, P 287 P 322 P 915 P 917 P 360 P 027, P 669 O 217, P 027, P 028, P 668, P 669 P 662 O 133 P 664 P 230 P 406, P 487 O 086 P 741 P 908 P 318, P 372, P 406, P 487 O 298 O 077, O 162 O 152, O 251, O 298 O 363 P 942 P 844 P 368, P 369, P 493 P 246 P 378, P 420 P 935, P 943, P 949, P 952 P 945 P 549 O 291 P 685, P 809 P 105 P 487 P 372 P 141 P 857 O 198, P 060 P 775 P 629 P 809, P 937, P 938 P 065 P 412 O 275 O 051 P 431 P 241, P 371 P 756 P 885, P 889, P 894 P 886 P 888 P 891 P 394 P 890

SOCCOL, V.T. SOHRABI, Y. SOLANO GALLEGO, L. SOLANO GALLEGO, L. SOLCA, M.S. SOLEIMANI, H. SOLER, R.C. SOMENZARI, M.A. SOONG, L. SOORI, T. SORDO, L. SOREMEKUN, S. SØRNES, S. SOSA BIBIANO, E.I. SOSA, L. SOSA, N. SOSSAU, D. SOTERIADOU, K. SOTERIADOU, K.S. SOTO, A.M. SOTO, M. SOTO, P.A. SOUB, A.C.S. SOUSA NASCIMENTO LEITE, K.S. SOUSA BATISTA, A.J. SOUSA, A. SOUSA, A.M. SOUSA, A.Q. SOUSA, B. SOUSA, F.D.G. SOUSA, G.R. SOUSA, L.C. SOUSA, L.O. SOUSA, M.C.S. SOUSA, O. SOUSA, P.B. SOUSA, R. SOUSA, S. SOUSA, S.A. SOUTHWORTH, D. SOUTO PADRON, T. SOUTTER, F. SOUZA NOGUEIRA, F. SOUZA FAGUNDES, E.M. SOUZA, A.C.N. SOUZA, A.P.A. SOUZA, B.L.S.C.

P 236 O 096, P 826 O 049, O 086, O 260 O 213 P 646 P 391 P 526 O 044 O 195, O 127 P 577 P 398 O 053, P 016 P 085 P 934 O 042 P 230 P 647 P 707 O 362 O 132 P 406, P 487 O 135 O 380 P 132, P 166, P 223 P 018, P 093, P 226, P 375 P 624 O 083 P 158 P 558 O 188 P 731, P 735 P 813, P 892 P 887 P 815 P 660 P 880 P 867 P 530 P 009 P 307 O 166, P 087 O 293, P 726 P 594 O 059 P 246 O 213 P 474 P 643 P 851 P 132, P 844 O 156

SOUZA, B.V.C. SOUZA, C. SOUZA, C.M. SOUZA, C.S.F. SOUZA, D.A. SOUZA, D.M. SOUZA, F.A. SOUZA, G.O. SOUZA, H. SOUZA, H.C. SOUZA, I.S. SOUZA, J.B. SOUZA, J.V. SOUZA, L.D. SOUZA, L.E. SOUZA, L.M. SOUZA, M.A. SOUZA, M.F.V. SOUZA, P.A.D.N. SOUZA, P.A.N. SOUZA, P.R. SOUZA, R.S. SOUZA, V.A. SOUZA, V.B. SOYZA, P. SPÄTH, G. SPÄTH, G.F. SPONTON, A. SRIVASTAVA, P. SRIVASTAVA, R. STABELI, R. STAGER, S. STAHL, H. STAHL, K. STAMPER, L.W. STAPLETON, J.T. STARK, O. STARKE BUZETTI, W.A. STASHENKO, E. STAUCH, A. STEINDEL, M. STEINKAMP, H. STELL, F. STEVENS, Y. STEVERDING, D. STIENSTRA, S. STILLE, K. STOBBE, N.S.

P 507 P 310 P 793 P 885, P 888, P 889, P 891, P 894 P 886, P 890 P 897 P 731 P 507 P 832 P 297, P 623 O 257, P 491 P 626 P 549 P 489 P 842 O 069 P 509 P 939 O 091, P 067, P 072 P 590 O 082 P 189, P 307 P 765 P 411 P 010, P 205, P 220, P 265, P 566 P 440 O 181 P 524 P 080, P 718 P 550 P 713 P 183 P 005 P 668 O 205, O 284, O 337 P 232 P 421 P 186 P 602 P 108 O 371 O 064 O 064 O 121 P 278 O 137, O 140, O 141, P 364 P 539 P 497 P 382 P 357, P 515, P 666, P 866 O 308, O 339 P 680 O 194, O 289 P 482 O 338 O 262 P 854

STOBER, C.B. STOCKDALE, L. STOLF, B.S. STRASSER, R. STRAUS, A.H. STREATFIELD, P.K. STRUB WOURGAFT, N. STRUCHINER, C.J. STUHLMANN, F. SUAREZ, J. SUAREZ, M. SUAREZ, M.S. SUAU, H.A. SUCRE, A.P. SUCUPIRA, M.V. SUDARSHAN, M. SULEMAN, M. SUMAN, S.S. SUMI, D.V. SUMOVA, P.

P 011 O 360 P 004, P 031, P 033, P 044 O 131 O 335 P 796 P 627 P 708 O 188 O 380 P 038 P 036 O 029 O 144, P 017 P 522 P 454, P 478, P 752 O 190 P 599 O 168 P 029 O 244

SUNDAR, S. SUNYOTO, T. SURANGO, B.P.S. SUZANNE, G. SVOBODOVA, M. SZELAG, E.A. TABBABI, A. TACCHINI COTTIER, F. TAFURI, W. TAFURI, W.L. TAHERI, 1.

TAHERI, T. TAIPALE, A. TAKAGI, H. TAKAHASHI, H.K. TAKELE, Y. TAKIYA, C.M. TALADRIZ, A. TALHARI, A. TALHARI, A.C. TALHARI, S. TALIGNANI, L. TALLY, J.D. TALVANI, A.

O 113 O 002, O 140, O 143, O 206, O 207, O 235, O 299, O 012, O 015, O 089, O 315, O 316, O 344, O 352, O 357, O 055, O 058, O 063, O 163, O 175, O 265, O 268, P 011, P 086, P 088, P 154, P 186, P 363, P 364, P 383, P 401, P 454, P 478, P 483, P 495, P 582, P 597, P 749, P 752 O 269 P 918 P 544 O 049, O 086, O 298 O 260 O 097 P 790 O 133, P 410 O 250 P 309, P 310, P 311, P 317, P 629 P 311, P 629 P 606, P 839 P 066, P 206, P 316 P 340 O 118, O 025, P 109 O 107 O 173 O 335 O 133 P 432 P 461 P 189 P 307 P 307 P 189 P 140 P 607 O 180

TAMAS, M.J. TANDON, R. TANDON,, R. TANIGUCHI, H.H. TANIWAKI, N.N. TAPADAR, J. TAPIA, F.J. TARALLO, V.D. TARTAROTTI, A.L. TASLIMI, Y. TATE, E.W. TAVARES, A.M. TAVARES, C.A. TAVARES, C.A.P. TAVARES, J. TAVARES, L. TAVARES, L.C. TAVARES, L.H.M. TAVARES, M.F.M. TAVARES, N. TAVARES, N.M. TAVARES, V.B. TAYLOR, M.J. TAYPE, C.A. TEESCH, L.M. TEIXEIRA NETO, R.G. TEIXEIRA CARVALHO, A. TEIXEIRA NETO, R.G. TEIXEIRA PINTO, L.A.L. TEIXEIRA, C. TEIXEIRA, D.E. TEIXEIRA, E.M. TEIXEIRA, F. TEIXEIRA, M.C. TEIXEIRA, M.J. TEIXEIRA, N.C. TEIXEIRA, P.C. TEIXEIRA, S. TEIXEIRA, S.C. TEIXEIRA, S.M.R. TEIXEIRA, S.R. TEIXEIRA, T.L. TEIXEIRA, V.N. TEKWANI, B.L. TELES RABELLO, A.L. TELES JUNIOR, E.P. TELES, C.B.G. TELES, M.

O 004 P 041, P 042, P 381 O 089 P 489, P 601, P 631, P 658, P 658, P 767 P 507 P 389, P 418 O 063 P 046, P 047 P 025, P 127, P 203, P 254 O 010 P 854 O 239, O 025 O 152 P 830 P 375 P 226 P 520 P 018, P 123, P 362, P 376 P 519 O 319 O 220, P 772, P 786 P 371 P 943 P 373 O 238 P 136 O 275 P 036 P 581 O 158 P 245 O 302 P 798 P 069, P 071 O 276, P 643 P 683 P 168 O 160, O 161, O 320, P 038 P 103 P 516 O 081, P 587, P 588 O 182, P 185 O 059 P 854 P 034, P 171, P 174 P 020 P 033 P 035 P 161, P 229, P 511 P 165 O 350 P 161, P 176, P 229, P 247, P 511 P 098 P 324 O 081 P 321 P 344 P 345 P 250

TELES, P.P.A. TELES, Y.C.F. TELLEVIK, M.G. TELLEZ, J. TELLO, A. TEMI SHIO, M. TEMPONE, A.G. TEMPONE, A.J. TENORIO, A.P.M. TEODORO, A.K.M. TEODORO, U. TERCEIRO, B.R.F. TERRAO, M. TERUYA, E.O. TERZIC, D. TESTASICA, M.S. TESTASICCA, M.C.S. TETIK, A.V. THAMMAPALO, S. THANGARAJ, K. THELLIER, M. THIES, S.F. THOMAZ SOCCOL, V.T. THOMAZ SOCCOL, V. THOMPSON, A. THOMSON, A. TIBURCIO, A.M.L. TIBURCIO, M.G.S. TILLEY, P. TIMMIS, J. TINOCO, N.A.B. TIVELLI, P. TIWARI, A. TIWARY, P. TODT, M. TOLEDO, C.R.S. TOLEDO, M.S. TOLEDO, R.S. TOLEZANO, J.E. TOLOUEI, S. TOMAS, A.M. TOMOKANE, T.Y. TONELLI, G. TONIN, L. TOPNO, R.K. TORALES, M. TORO LONDOﾃ前, M.

P 630, P 931, P 932 P 440 P 230 P 515 P 766 O 338 P 482 P 370, P 409, P 641 P 351, P 461, P 505 P 367, P 389, P 390, P 464, P 612 O 246 P 488, P 528, P 594 P 911, P 912 P 236, P 738, P 849, P 858, P 862 P 164 O 348 P 131 O 013, P 426 P 145 P 253 O 321 P 662 P 137 P 457 P 754, P 755 P 098, P 738 P 441, P 444, P 742, P 761, P 776 P 596, P 636, P 642 P 450 P 145 P 214 P 462, P 781 P 803, P 807 P 476 O 340 P 210 P 054, P 056 P 114 O 063 P 154, P 495, P 582, P 597 O 209 P 756 O 335 P 829 P 631, P 857 P 395, P 489, P 601, P 658, P 767 P 299, P 300 O 070, O 059, P 262 P 471 O 182, O 188, P 185 P 394 P 736 P 146, P 291, P 793 P 798 P 075 O 056, O 061, P 599 O 270 P 529

TORO, M.A. TORRACA, T.S.S. TORRELLAS, A.M. TORRENT, M. TORRES SANTOS, E.C. TORRES, A.R.A. TORRES, B. TORRES, C. TORRES, F.D. TORRES, M.S. TORRES, O.D.S. TORRES, T.M. TORRES, U.P.S. TORRICO, M.C. TOUMI, A. TOVAR, B. TOZ, S.Ö.

TRAJANO, L.A.M. TRANNIN, M.A. TRAUB CSEKÖ, Y.M. TRAVI, B.L. TRENTIN, B. TRINCONI, C.M. TRIPATHI, C.D.P. TRIVEDI, V. TRONCO, C.T. TROVATTI, E. TRUJILLO, J.J. TRUPPEL, J.H. TSEGAYE, I. TSIGANKOV, P. TSUGE, C. TSUNASHIMA, R. TULADHAR, R. TUMA, K.S. TUMBAICO, J.W.T. TUNA, G.E. TUNON, G.I. TUON, F.F. TURCO, S.J. TURGAY, N. TURNER, J. TUTTERROW, Y. UBEDA, J. UCHOA, F.C. UEDA NAKAMURA, T. ULIANA, S. ULIANA, S.R.B.

O 307 P 090, P 164 O 259 P 475 P 120, P 408, P 421 P 266, P 267 P 222 O 196 P 659, P 850 P 479 P 657 P 285 P 830 P 438 O 125, P 039, P 305, P 306, P 916 P 254 O 135 P 585 O 136, P 073, P 119 O 246 P 447 O 345, P 059 P 419 O 089, O 120, P 043, P 388 O 104, P 130 P 473, P 477 P 433 P 904 P 738 P 750 O 346 O 179 P 313 P 094 P 952 P 918 O 321 P 740 O 168, P 435 O 075 P 117, P 287 O 158 O 092, O 027 P 336 P 489 P 625 O 282 P 405 P 404, P 418, P 419, P 473, P 477

ULLMAN, B. UMAÑA PEREZ, A. UNCOS, A. UNRAU, P. UNVER, A. UPEGUI, Y.A. URANW, S. URBINA, J.A. URIBE, S.I. UTGES, M.E. UZONNA, J.E. VALADARES, D.G. VALDERRAMA, A. VALDERRAMA, C.H. VALDES, A.C. VALDIVIA, H.O. VALDIVIA, H.V. VALDIVIESO, E. VALE, M.M. VALENCA, W.O. VALENCIA, A. VALENCIA, B. VALENCIA, B.M. VALENZUELA, J. VALENZUELA, J.G. VALERIO BOLAS, A. VALETE ROSALINO, C.M. VALLEJO, A. VALVERDE, J.G. VALVERDE, J.L.G. VAN DER AUWERA, G. VAN DER GEEST, S. VAN DER VEER, C. VAN GRIENSVEN, J. VAN LUENEN, H.G. VAN WYNSBEGHE, N. VANAERSCHOT, M. VANEGAS, O. VANERSCHOT, M. VANWALLEGHEM, G. VARGAS, D.A. VARGAS, E. VARGAS, M.L.F. VARIKUTI, S. VARJAO, J.R. VASCONCELOS, A.C. VASCONCELOS, M.F.

O 132 O 325 O 038, P 485 O 054 P 117 O 280, P 327, P 469 O 078, O 111, O 357, O 067 P 352 O 252, P 899 P 951 P 773 O 373 P 362 P 895 O 008 P 743, P 744 O 351 P 680 P 835 P 429, P 494 P 705 P 492 O 196 P 144, P 438 P 397, P 535 O 159, O 025, O 026, P 038, P 844 O 161, O 318, P 958 O 160, O 320, P 029 O 331, P 772 O 237, P 037 P 090, P 164 P 243 P 255, P 273, P 286 P 918 O 254, O 255, O 290, P 061, P 062, P 438 O 098 P 151 O 228, O 262, O 263, O 269 O 107 P 647 O 002, O 143, O 111, O 344, P 382, P 383, P 384, P 385, P 386, P 672, P 679 P 017 O 113 P 101 O 147, P 089, P 100 P 836 P 532 O 339 P 820 P 051 P 408 P 421

VASCONCELOS, R.O. VASCONCELOS, T. VASQUEZ, D. VASQUEZ, G.M. VASSALLO, M. VAUCHY, C. VAZQUEZ, M.A. VELAND, N. VELASQUEZ, E. VELASQUEZ, L.G. VELAZCO, E. VELAZQUEZ, S. VELDHUIS, S. VELET, M. VELEZ, A. VELEZ, I. VELEZ, I.D. VELO, E. VELOSO, V.M. VELURU, N.K. VENDRAME, C.M.V. VENDRAME, C.V. VERA, H. VERAS, L.M.C. VERAS, P.S. VERAS, P.S.T. VERCOSA, B.L.A. VERCOZA, B.R.F. VERDIANO, V.A. VERDONCK, K. VERGARA, D. VERMA, N. VERMELHO, A.B. VERMONT, M. VERONESI, F. VERSTEEG, I. VERWEIJ, J. VIANA, J.B.M. VIANA, K.F. VIANA, S.M. VIANNA, E.N. VICTOIR, K. VICTORIA, C. VIDAL, M.L. VIDAL, R.O. VIDOTTO, O. VIEGAS, R.P.

P 177, P 190 P 268 P 794 O 164 P 680 P 221 O 118 P 766 O 254 P 040, P 380 P 031 P 259 O 307, P 529 O 023 P 272 O 011, O 196, O 380, P 904 O 202 O 364 O 011, O 280, O 283, O 361, O 365, O 196, O 252, O 164, O 380, P 327, P 328, P 469, P 589, P 899, P 904, P 910 O 254 P 843 P 533, P 816 O 104 P 590 P 294 P 680 P 561 O 305 P 074, P 326, P 522, P 524, P 526 O 036, O 181, O 044, P 106, P 391 P 051 P 399, P 414 P 813, P 815 O 267, O 269, P 144 O 011 O 061 O 108, O 057, O 261 P 445 P 460 P 604 O 060 O 255 P 293, P 296, P 608, P 609, P 614, P 617 P 197, P 199, P 292 P 034, P 171, P 174 P 816, P 954 P 663 P 863 P 293 O 030 P 829 P 788

VIEIRA BRANQUINHO, H.C. VIEIRA FILHO, J.N. VIEIRA GONCALVES, R. VIEIRA, A.T.P. VIEIRA, E. VIEIRA, E.L.M. VIEIRA, P.M.A. VIEIRA, P.S. VIEIRA, R.F.C. VIEIRA, S. VIEIRA, T.S.S. VILA NOVA, N.S. VILACA, R. VILAS, F. VILELA, M.L. VILLANUEVA LIZAMA, L.E. VILLELA REZENDE, G. VILLELA, E.V. VIOL, M.A. VISHWAKARMA, P. VITAL, W.C. VITALE, F. VITALE, M. VIVANCO, B.C. VIVARINE, A.C. VIVARINI, A.C. VIVERO, R. VIVERO, R.J. VIVEROS, R.J. VLKOVA, M. VOGAS, G.P. VOJTISKOVA, J. VOLF, P. VOLFOVA, V. VOLKOVA, V. VON STEBUT, E. VONATSOU, P. VOTYPKA, J. VOTÝPKA, J. VOUGOGIANNOPOULOU, K. VOULDOUKIS, I. WACKWELLA, H. WAGHABI, M.C. WAGNER, G.K. WALLACE, J. WARBURG, A. WARD, R.J.

P 321 P 319 P 551 O 233 P 883 P 250 O 050 O 091, P 067, P 072, P 714 P 557 P 395, P 852 P 786 O 301, O 304 P 622 P 786 P 766, P 769 P 933 P 647 O 276 P 217 P 402 P 596, P 642 P 522, P 714, P 900, P 903 O 187, P 064, P 400, P 693, P 837 P 050 P 156, P 172, P 175 P 222 P 183 O 037 O 196, O 380 O 252 P 951 P 910 P 029 P 393 O 294 O 086 O 049 O 216, O 244, O 245, O 248, O 249, O 251, O 298, O 318, P 029, P 717 O 248, P 029 O 049, O 086, O 260 O 333 P 901 O 216, O 248, O 249 P 717 P 406 O 214, P 068 P 670 P 065 P 482 P 450 O 216, O 248, O 249, P 029, P 751 P 557

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ZINI, R.M. ZORRILLA, V.O. ZULETA, M. ZULUAGA, J.J. ZUﾃ選GA, C. ZUQUE, F.R.S. ZUQUE, F.T.S. ZUQUE, M.A.S. ZWETSCH, A.

P 857 P 680 O 196 O 365 O 380 P 745, P 799 P 745, P 799 P 745, P 799, P 873 P 870, P 933

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