Veterinary College, Bengaluru Monthly e-Bulletin
Newsletter Date : 31st May 2017
Volume No: 06 Issue: 05
Rathnamma, D*., Usharani, J., Kavita, G**., Veeregowda, G*. Viswas K. N***. *Department of Veterinary Microbiology, Veterinary College Hebbal, **Institute of Animal Health and Veterinary Biologicals, ***NIVEDI, Email: rathnarohit@gmail.com Introduction: Clostridia are Gram positive, rod shaped, obligate anaerobic endospore forming organism occurring worldwide and are ubiquitous in the environment. This genus covers wide range of hosts usually humans, ruminants, horses and poultry. Consuming contaminated or intoxicated feed or water is the major route of infection. Most clostridia produce toxins which are the major virulence factors for the pathogenesis of infection. Other predisposing factors such as changes in dietary ingredients, severe stress, injury, other infectious agents, coccidiosis, or immunosuppressive infections such as infectious bursal disease or chicken infectious anemia, are responsible for the establishment of anaerobic clostridial infections. As a result, clostridial diseases are responsible for great economic loss to poultry industry either by death or due to poor production and growth rate. Major clostridial infections which affect poultry include – 1. Ulcerative enteritis caused by Clostridium colinum; 2. Necrotic enteritis or gangrenous dermatitis by C. perfringens, C. septicum and S. aureus 3. Botulism by C. botulinum. There are some sporadic case reports of C. chauvoei, C. difficile, C. fallax, C. novyi, C. sordellii and C. sporogenes infections in poultry. Ulcerative enteritis or Quail disease Ulcerative enteritis is an acute, highly contagious disease of quails and poultry caused by Clostridium colinum. It is characterized by ulcers of the intestines and caecae with 100% mortality in quails while in chickens only 10%. Young birds of chicken 4-12 weeks, turkey 3-8 weeks, and quail 4-12 weeks of age are more frequently affected than adult birds. The organism being fastidious requires enriched media with anaerobic growth condition. Major clinical signs of ulcerative enteritis include listlessness, retracted neck, drooping wings, partially closed eyes, ruffled feathers, diarrhea, anaemia and watery white faeces in quail.
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Botulism (Limber neck or Western duck sickness) Botulism is an intoxication prevalent worldwide caused by a preformed neurotoxin (exotoxin) of Clostrdium botulinum type C. The disease is characterized by the progressive flaccid paralysis. There is worldwide distribution of botulism affecting wide range of free range as well as confinement reared poultry, wild birds, avian and mammalian predators and scavengers. An estimated 10 to 100 thousand birds die of botulism annually and in some large outbreaks, a million or more birds may die leading to huge loss. Among all the bird types, ducks are the one which gets affected more often. In case of chickens, flaccid paralysis progress from legs, wings, neck, and eyelids. Birds are found sitting and are reluctant to move, wings drooped, paralysis of the neck, when eyelid get paralyzed bird look comatose and dead. Finally death is due to cardiac and respiratory failure. Gangrenous dermatitis (necrotic dermatitis, gangrenous cellulitis, gangrenous dermatomyositis, avian malignant edema, gas edema disease, wing rot or red leg) Gangrenous dermatitis is a disease affecting fattening and growing birds of 4 to 8 weeks of age. Etiological agent/s of the gangrenous dermatitis include/s C. septicum, C. perfringens or S. aureus, either singly or in combination. The disease is characterized by necrotization of different skin areas and a severe cellulitis of the subcutaneous tissue revealed as dark, moist areas of skin with exposed underlying muscle areas. Birds exhibit varying degrees of depression, incoordination, inappetence, leg weakness, ataxia and death within 24 hrs. The affected birds have dark, moist areas of skin devoid of feathers, overlying wings, breast, abdomen, or legs with extensive blood-tinged edema, with or without gas (emphysema), is present beneath affected skin. Necrotic enteritis Necrotic enteritis is an acute toxico-infectious disease caused by toxin released by C. perfringens (C. welchii) Affecting poultry especially broiler chickens (2-5 wk old), turkeys (7-12 wk old) and ducks raised on litter but can also affect commercial layer pullets raised in cages. The organism commonly resides in the intestinal tract of the host. Hence, through faeces the organism reaches and contaminates the soil, litter, feed, water and surrounding environment. Alteration in the intestinal microflora or from a condition that results in damage to the intestinal mucosa (eg, coccidiosis, mycotoxicosis, salmonellosis, ascarid larvae, immune suppression), high dietary levels of animal byproducts that promotes excessive bacterial growth and toxin production or slows feed passage rate and provides anaerobic condition could promote the multiplication of bacteria and production of excessive amount of alpha toxin results in the occurrence of necrotic enteritis. The disease may be clinical or subclinical. In clinical cases, there will be sudden increase in flock mortality, often without premonitory signs, although wet litter. Gross lesions are confined to small intestine, primarily jejunum and ileum. Intestines are often friable and distended with gas.
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Diagnosis of clostridial diseases Diagnosis is by history and clinical signs of the disease. Gross lesions and microscopic characteristics of the affected organs. Isolation and identification of causative agent support the diagnosis. Confirmation is by isolation and identification of the causative agent. PCR based on 16S rRNA sequence, multiplex PCR targeting the different toxins (C. botulinum, C. perfringens) ELISA, direct fluorescent antibody test is highly specific and confirmative test. Apart from above tests general diagnosis and molecular PCR, Mouse bioassay test, Colony immunoblot assay, Fluroscent antibody technique, antigen-capture ELISA, commercial lateral flow assay, can be used for detection of botulinum toxin in contaminated material. Toxin neutralization test either by invivo test in mice or by invitro test is gold standard for toxin typing of C. perfringens. Control measures Disease control in poultry farm is difficult, but prevention of outbreak can be taken care by removing the dead birds often, maintaining house environment that is as dry as is practical (environments that are too dry will encourage respiratory diseases), keeping equipment as clean as possible (particularly open-type waterers) and preventing other animals from entering the house (domestic animals, wild animals, birds and humans). Managing the clostridial disease during outbreak is very important as these are spore forming bacteria, complete eradication including spores, disinfecting the area, clean out of poultry stock, use of probiotics, vitamins etc., helps in controlling the clostridial infections. Total clean out of farms, followed by thorough cleaning and disinfection of the house and floor, using large amounts of water mixed with a phenolic disinfectant (1500 gallons per 20,000 ft 2) is helpful in prevention of the disease. Treating the floor with salt at 60-100 pounds per 1,000 ft2 prior to placement of bedding material also has decreased the incidence of gangrenous dermatitis on problem farms. Management procedures to improve litter condition, reduce moisture and bacterial levels in the environment, and minimize trauma are useful adjuncts to treatment
Nishanth. C, Mohan H. V And Leena G Dept. of Veterinary Public Health And Epidemiology, VCH, Bengaluru. (Email: nishanthvet @gmail.com) Introduction Vector-borne Disease (VBD) are those infections that are transmitted by the bite of infected arthropod species, such as mosquitoes, black flies, triatomine bugs, ticks etc. Around the world, Vector borne disease accounts for more than 17% of all infectious diseases. One-sixth of illnesses worldwide is due to VBD resulting in the annual mortality of more than one million. India is endemic for six major vector-borne diseases namely dengue, malaria, filariasis, chikungunya, Japanese encephalitis, and visceral leishmaniosis in human beings. The major vector-borne diseases of animal importance in India are trypanosomiasis, blue tongue and babesiosis. However recently Crimean – Congo Haemorrhagic Fever
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and West Nile Fever antibodies have also been reported in animals. Vector borne disease have re-emerged and spread to new parts of the world during past two decades. Zika virus fever Zika viral disease is a mosquito borne viral disease, caused by a single stranded, positive sense RNA virus belonging to the family Flaviviridae, genus Flavivirus. The disease is transmitted to humans by the bite of infected mosquitoes of Aedes species. Reports of transmission of disease through intra uterine route (congenital/vertical) and also sexual route has also been noticed. According to WHO, Zika virus (ZIKV) transmission has been documented in 59 countries reporting the autochthonous transmission of the disease. Zika virus was first isolated in 1947 from a rhesus monkey in the ZIKA forest of Uganda, followed by its first report in humans in 1952. The first reported, official epidemic took place in island of Yap, Micronesia, in 2007. From then on the disease has spread across the world.
Epidemiology and transmission of ZIKA virus
The incubation period of the disease is unknown and the symptoms lasts for 2-7days. The disease is manifested clinically from asymptomatic cases to an influenza like syndrome associated with mild fever, non-purulent conjunctivitis, headache, maculopapular rashes and less frequently, retro-orbital pain, diarrhoea and abdominal pain. Microcephaly is the outcome in new born kids. Biological confirmation of ZIKV infections is based on detection of virus RNA in serum by using RT-PCR. There is neither vaccine to prevent nor specific anti-viral treatment available. Treatment is directed primarily at relieving symptoms using anti-pyretics that can reduce fever and analgesics that relieve pain. Prevention and control is by, Mosquito control and pregnant women should avoid travelling to place where ZIKV is circulating. Dengue Fever Dengue fever (DF) classical dengue fever (DHF) and dengue shock syndrome (DSS) are the most important emerging mosquito borne disease of human beings. Dengue fever is caused by infection with four serotypes such as DEN-1, 2, 3 and 4, which are closely related to each other antigenically. The virus belong to the family Flaviviridae and genus Flavi virus. Dengue fever incidence has increased in numbers of infected individuals over the period and is characterized by occurrence of high fever, sever body ache and intense headache. It is transmitted through the bite of the female mosquitoes belonging to the genus Aedes. Incubation period varies from 5-6days, but however it can extend up to 10 days. The classical dengue fever is a self-limiting disease and may not prove fatal. However, the other types (DHF and DSS) can prove fatal if prompt treatment is not started. There is no treatment including vaccines and drugs.
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Supportive treatment such as anti pyretics and hydro therapy can be given to the patient. If any of the symptoms are indicative of DHF and DSS develop, rush the patient to the hospital and may require transfusion of fluids or platelets. Prevention of breeding and protection from the biting of the Aedes mosquitoes could be the best control measures. It is important to remember that the Aedes mosquitoes usually bite during day time and hence one should take necessary measures accordingly. Chikungunya fever Chikungunya fever is a mosquito borne viral disease, caused by a single stranded, positive sense RNA virus belonging to the family Flaviviridae, genus Flavi virus. It is transmitted to human beings by the bite of infected Aedes mosquitoes. This infection can cause a debilitating illness, most often characterized by fever, headache, fatigue, nausea, vomiting, muscle pain, rashes and joint pain. Incubation period varies from 2-12 days but usually it is 3-7 days. Symptoms and signs are Chikungunya are similar to dengue except the mortality rate is very high in dengue. Protection from mosquito bite and supportive therapy are only the helpful guidelines and preventive measures.
Chikungunya virus transmission cycle
Japanese encephalitis Japanese encephalitis (JE) is the largest worldwide cause of epidemic viral encephalitis. The first reported outbreak of the disease occurred in Japan causing a huge mortality in human beings. Virus was isolated for the first time from a post-mortem case of human brain in Japan in 1933. The disease was clinically diagnosed for the first time in India in 1955 at Vellore, North Arcot district of Tamil Nadu. In India, JE is considered mainly as a paediatric problems characterized by severe encephalitis with compact clinical course, high mortality, and high disability rate in survivors. Japanese encephalitis virus is a negative sense, single stranded enveloped RNA virus belonging to genus Flavi virus, family flaviviridae. The disease has re-emerged causing recent outbreaks in different parts of India that includes Karnataka (2005), West Bengal (2009), U.P (2011), Delhi (2011), Bihar (2011) whose intensity has alarmed the public health agency of the country. The disease is considered to be an emerging public health problem in southern states namely Kerala, Tamil Nadu (TN), Andhra Pradesh and Karnataka. Outbreak of the disease occurs throughout the year with the higher risk after rainy season and the prevalence of disease is more in paddy growing areas NVBDCP (2012). New born and young children’s are at risk of contracting the disease. Japanese encephalitis virus is transmitted to humans through mosquito bites. Pig’s acts as an important amplifiers of the virus producing high viremia’s which infect mosquito vectors.
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Mosquitoes belonging to genus Culex, mainly Culex tritaeniorhynchus transmits the virus upon biting human beings. The natural maintenance reservoir for JE virus are birds of the family Ardeidae (herons and egrets), although they do not demonstrate clinical disease they do generate high viremias upon infection. Horses are the primary affected domestic animals of JE though essentially a dead-end host; other equidae (donkeys) are also susceptible. Humans are vulnerable to the disease and this disease is a primary public health concern in Asia; humans are considered as dead-end host, no person-person or animal-person transmission has been reported. Incubation period in animals varies from 8 to 10 days showing varied neurological lesions such as impaired locomotion, stupor, teeth grinding, Blindness, coma, death (rare). Incidence of mummified foetuses and infertility has also been noticed in Pigs. In man a Prodromal Stage lasts for 1 to 6 days. An Acute encephalitic Stage begins by third to fifth day, the symptoms include fever convulsions, , stiff neck, muscular rigidity, tremors in fingers, tongue, eyelids and eyes and a Late Stage is characterised by the persistence of signs of CNS injury such as, mental impairment, , paresis, speech impairment, epilepsy, abnormal movements and abnormal behaviour.
Transmission cycle of Japanese Encephalitis
Diagnosis is through detection of JE virus-specific IgM antibody in CSF or serum by an IgM-capture ELISA specifically for JE virus, detection of JE virus genome in serum, plasma, blood, CSF, or tissue by RT-PCR or an equally sensitive and specific nucleic acid amplification test. Treatment of JE is essentially supportive. Measures to be taken for the control of vector borne disease: 1. Early Case Detection and Treatment. 2. Vector Control includes: Reduction of breeding source for larvae- use of larvicides, draining mosquito habitats. Reduction in man-mosquito contact- use of mosquito repellents, bed nets. Control of adult mosquitoes – using insecticide. 3. Pig control: segregation, slaughtering and vaccination of the pigs wherever, they act as an amplifier host. 4. Vaccines- vaccines are available in market for the control of few of the vector borne diseases such as JENVAC for the control of Japanese encephalitis in children’s. The use of these vaccines should be
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propagated. 5. Integrated Disease Surveillance in humans – use of some of the commercially available kits for the diagnosis of JE (Xcyton kit), Dengue (PanBio kit) and other vector borne disease can be used as a part of integrated disease surveillance program in India. 6. Integrated Disease Surveillance in animals- Japansese encephalitis elisa kit for swines have been recently developed by a team of scientist, Division of veterinary public health, IVRI for the screening of the pigs infected with JE as they serve as the amplifying host. Similar approaches has to be developed in other vector borne diseases.
B. Kamal Hasan1 and M.A. Kshamma2 . Contract teacher1 , Associate Professor2, Veterinary College, Bengaluru. Email Id: kamalmicrobiology@gmail.com A proper investigative approach to dermatologic cases should include the following 1) History 2)Physical examination 3) Laboratory and other diagnostic techniques ( Hematology ,Biochemistry, biopsy). The skin diseases can be mainly classified in to the following categories : 1. Bacterial skin disease ( Pyoderma) : these include mainly surface pyoderma and deep pyoderma . 2. Parasitic skin disease: these may be ticks, lice, mites , fleas, flies helminth parasites (Ancylostoma caninum), protozoa ( Leishmania canis), rickettsia. 3. Fungal skin disease : include ringworm, mycetoma sporotrichosis, histoplasmosis, cryptococcosis, blastomycosis. 4. Viral skin disease: include canine distemper. 5. Seborrhoea: canine seborrhea and other sebborhoeic syndromes include tail gland Hyperplasia. 6. Immunological skin disease: include the Type – I, Type –II, Type- III, Type- IV hypersensitivities, atrophy, food allergy, auto- immune disorders. 7. Physical skin disease: like nasal solar dermatitis. Burns and frost bite also affect skin. 8. Chemical skin disease : may be due to chemicals like strong acid and alkali. 9. Tumour condition of the skin like papilloma, sweat gland tumour, sebaceous gland 10.Tumour, mast cell tumour (mastocytoma) 11.Physical skin disease like nasal solar dermatitis. Burns and frost bite also affects skin. 12.Chemical skin disease may be due to chemicals like strong acids and alkalies. 13.Inherited skin diseases like hypotrichosis, dermatomyositis. 14.Nutritional skin diseases may be result of a deficiency of Protein, essential fatty acid, vitamins 15.Skin conditioning due to poisoning due to snake bite, bee sting. Canine Pyoderma: The main causative microorganisms include staphylococcus epidermididis, Corynebacterium, Clostridium spp. It affects all age groups. symptoms include hot spot, wet eczema. Treatment may include cleaning with shampoos, such as 2.5 % benzyl peroxide. Parenteral treatment with Antibiotics may also be advocated with surgical drainage in some cases. Viral skin disease : In some cases of distemper, there is hyperkeratosis of nose and foot pads. Prevention is achieved by vaccinating the animal at 4,8, 12 and 16 weeks of age and annual
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vaccination is recommended. Treatment includes use of broad spectrum antibiotics for prevention of secondary bacterial infection, application of antiseptic lotion on the pustules and symptomatic treatment for systemic disorders. Endocrine skin disease: Hypothyroidism is the commonest endocrine disorder in dog and affects any age but mostly between 6 and 10 year of age. Some breeds are reported to be predisposed like Doberman, Boxer. There may be bilateral symmetrical alopecia, hyper pigmentation, thickening of skin and scaling. Diagnosis is based on history, physical examination, laboratory investigation and findings and drug of choice is thyroxin at the rate of 10-20 mg/kg given two times a day. Immunological skin disease: May show acute onset wheals, hairs in affected areas stand up and Pruritis is observed. Diagnosis may done by history , physical examination and to treat, eliminate the known cause. adrenaline 1 :1000 may be given intravenously. glucocorticosteroids may be given in the case of utricaria. Antihistamines, antibiotics are given to prevent secondary bacterial infection and recently some enzyme and antiproteinase are of considerable value. Inherited skin disease: The specific skin disease may be diagnosed by history, pedigree, physical examination, histopathological examination of the biopsy material. once a diagnosis of a inherited skin disease is made, then the owner should be intimated that the problem is incurable and one should avoid trauma in such cases, such animals should not be bred. Skin conditions due to poisoning: In case of snake bite, first the wound is cleaned by clipping of hairs from wound area. Linear incision is given in many cases, polyvalent antiserum may be given by intravenous route and a little anti venom may be injected at the site of bite. Corticosteroids and antibiotics are given. In case of string bite, there are utricarial weals on skin. Wash with washing soda, and antihistamines are given to combat allergic reaction.
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Rijin Shankar,1Dr. Ranjith. D and 2Dr. Viswanath S College of Veterinary and Animal Sciences, Pookode, Kerala, 2 Veterinary Officer, Department of AH and VS, Karnataka, Email: ranjith946@gmail.com Introduction Parasitic infestations are one of the major constraints for profitable animal rearing in tropical and subtropical Countries including India. Gastrointestinal parasites cause considerable global economic losses as a consequence of reduced weight gain, digestive disturbance, lowered production, impaired reproductive performance, condemnation of affected organs and mortality. The economic losses are closely associated with the extent to which the pathogenic effect of helminth infections influences the production of the individual host. These may vary considerably from clinical disease including mortality to chronic production losses. Control of helminths relies almost exclusively on a limited number of synthetic anthelmintic drugs. The limitations of this reliance on chemotherapy are the threat of parasites developing resistance to drug treatment and the cost of drugs for small-scale farmers in developing countries and for some helminths, lack of efficacy of current available drugs. Although until recently the majority of the evidence on the antiparasitic activity of medicinal plants was anecdotal and lacked scientific validity, there is currently an 1
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increasing number of controlled experimental studies that aim to verify and quantify anthelminthic plant activity. There are indeed a large number of plants whose anthelmintic activity has been demonstrated under controlled experimentation, either through feeding the whole plant or administering plant extracts to parasitized hosts. The most important predisposing factors of helminth infections are grazing habits, climate, nutritional deficiency, pasture management, immunological status, vector, presence of intermediate host and the number of infective larvae and eggs in the environment. The effect of helminth infections is determined by a combination of factors, of which the varying susceptibility of the host species, the pathogenicity of the parasite species, the host/parasite interaction and the infective dose are the most important. It is impossible to draw general recommendations to control parasitic diseases due to varied geo-climatic conditions and methods adopted for rearing the livestock in the country like India. In view of increasing incidence of anti-parasitic drug resistance in animals, there is an urgent need to design sustainable parasite control strategy which must include on the host as well as off the host control measures to harvest the maximum productivity from the animal for an indefinite period. Herbal Dewormers: Some common botanical Dewormers include garlic; wormwood (Artemisia spp.); wild ginger or snakeroot; goosefoot; conifers (pine, spruce, or fir); mustard and castor oil; squash or pumpkin seeds; carrot and fennel seeds; pyrethrum (plant extract from Chrysanthemum) etc. The use of crude neem leaf powder (500mg/kg) against bovine strongylosis found to be beneficial. The extracts of Eucalyptus globulus which contains tannins and flavonoids and Cynanchum komarovii seeds with Verticillium dabliae can be used effectively against helminths. Clerodendrum colebrookianum walp (Lamiaceae) which is a perennial shrub can be used for treatment against tapeworm infections. Boiled Nutmeg water, Immature areca nut powder, pumpkin seeds, papaya and gentian Herbal Combinations includes: 1. Ajowan + Ginger + Black pepper 2. Beetle leaves + garlic +ginger + turmeric + cumin seeds 3. Aloe vera + Turmeric + Tamarindus indica + Jaggery 4. Ocimum sanctum+ honey+ ginger+ pepper 5. Cumin seeds (25 gms), Mustard (10 gms), Pepper (5 gms), Garlic (5 Nos), Leucas aspera (One hand full), Neem leaves (One hand full), Banana stems (100 gms), Bitter Gourd (50 gms), Jaggery (100 gms). Advantages: 1. Reduced risk of side effects: Most herbal medicines are well tolerated by the animals, with fewer unintended consequences / side effects than pharmaceutical drugs. 2. Effectives with chronic conditions: Herbal medicines tend to be more effective for long-standing health complaints that don't respond well to traditional medicine. 3. Lower cost: Herbs cost much less than prescription medications and usually considered as a boon to poor farmers.
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4. Widespread availability: Yet another advantage of herbal medicines are their availability. Herbs are available without a prescription. Plants and their combination
Helminthic infection in animals
Butea frondosa
100% lethal to earthworms
Ananas sativus + Embellia ribes + Macuna prurita + Melia azedarach
Taenia canina and Paramphistomum cervi
Carica papaya
Ascaris lumbricoides, Ascaridia galli
Cucurbita mexicana (Cucurbitaceae) seeds
Moniezia expansa, Fasciolopsis buski, Ascaris
Piper betle
Anticestodal activity
Cucurbita pepo + Calotropis gigantean, Juglans regia + Momordica charantia + Musa paradisaca + Scindapsus offcinalis
Haemonchus contortus of goat
Helleborus niger + rhizomes of Zingiber offcinale + seeds of Carum copticum + Agati gratifola + Mangifera indica.
Ascaris species
Caraca papaya + Sapindus trifoliatum (Sapindaceae) + Butea frondosa + Momordica charantia
Ascaridia galli worms
Boswellia serrata (Burreaceae) + Cinnamomum tamala
Anti-tapeworm activity
Conclusion: It is of prime importance, in both socioeconomic and scientific perspective to extrapolate the exuberant resource of Ethnoveterinary knowledge prevailed in the local healers of our country by documenting and scientifically validating the practices. The advantages of the herbal medicines includes as it is cheaper, more sustainable, easily available and reliable alternatives to synthetic drugs. Furthermore, when put into production locally, they can reinforce the income and status of local inhabitants. However, there are problems connected with the use of herbal medicine, the largest being the lack of scientific evaluation. Such evaluation is the most important step, once information about indigenous uses of medicinal plants has been collected. The most effective path for obtaining such knowledge is the ethnobotanical approach, which assumes that the indigenous use of plants indicate the presence of biologically active constituents in the plants. Herbal medicine may very well become a pioneer in turning local knowledge into global knowledge, through the recognition of local practices as an indispensable source of sustainable development for both people and the environment all over the world. This of course does not mean that traditional medicine should take over western medicine, but rather that we may benefit from a harmonious balance of both.
Usharani, J., Kavita, G*., S Isloor**., Rathanamma, D**., Viswas K.N*** *Institute of animal health and veterinary Biologicals, **Department of Veterinary Microbiology, Veterinary college Hebbal, ***IVRI, Bareilly, Email I.D: rathnarohit@gmail.com Lateral flow immunoassay is one of the rapid diagnostic technique which can be used at the point of care using any crude clinical sample containing the infectious agent. Lateral flow device is simple, compact, easily portable hand held prefabricated strips that do not require external reagents for results. The lateral flow assay is the extended principle of latex agglutination test. Lateral flow assay is not only
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qualitative but also semiquantitative. Key principle of lateral flow assay Includes the movement of the test analyte along the various zones of the polymeric material by capillary action. The analyte of interest interacts with the specific ligand/s on pre-engineered reporters and the complex of analyte and reporter move towards the test and control zone where molecules have been attached which exert more or less interaction with the test analyte. Based on the test formats whether sandwich or competitive the positive color signal is formed at both test and control zone or only at the control zone respectively (Figure 2 and 3) Figure 1: lateral flow format for immunoassay Sample applica-
Conjugate pad
Test zone
Control zone
Wick pad
Backing material
Figure 2: Sandwich lateral flow assay format results read out Positive
Negative
Figure 3: Competitive lateral flow format read out Positive
Negative
Lateral flow assay for the detection of some infectious diseases/toxicosis like Anhrax, Brucellosis, Dengue, Leptospirosis, FMD,Rabies, CD, Canine Parvoviral gastroenteritis, Syphilis, Botulinum toxicosis, Staphylococcal enterotoxin B (SEB), Ochratoxin A (OTA), Aflatoxin B1 Merits of the lateral flow immunoassay system (LFIA) Relative ease of manufacture – equipment and processes already developed and available Stable – shelf-lives of 12–24 months often without refrigeration Ease of use: minimal operator-dependent steps and interpretation
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It can work well with the multiple samples in a small volume amounts It can be a quantitative assay if we are integrating with reader system Economical, easy and involves less time for development Market presence and acceptance – minimal education required for users and Regulators Drawbacks/limitations of LFIA Qualitative or semiquantitative High concentration of protein sample compared to the other immunoassays Restriction of sample volume It is only for single use Sensitivity of lateral flow assay is much less than the ELISA Should maintain the integrity of the membrane and carefully preserve the membranes till use
Gaikwad S S., Rathnamma, D., Usharani, J., Kavita, G. Department of Veterinary Microbiology, Veterinary College Hebbal. Email: rathnarohit@gmail.com Introduction: Rapid and cost effective diagnostics are highly desirable for detection of veterinary pathogens. For emerging and re-emerging zoonotic viral pathogens which have public health significance, rapid onsite diagnosis is most desirable. Biosensors offer alternatives to conventional diagnostic methodology for pathogen detection. Antibody based biosensors are a class of biosensors which provide high specificity and sensitivity. This technology has huge potential due to recent developments in the area of antibody generation, microfluidics nanotechnology. Infectious diseases of animal not only account for economic losses due to increased treatment costs, loss of production, morbidity and/or mortality but, also due to public health risk these pathogens pose. The challenges are greater in case of viral infections which are relatively difficult to detect and treat. These pathogens adapt themselves for survival, by mechanisms such as mutation, recombination, reassortment, host switching and acclimatizing to new environment. The diseases with huge economic repercussions, e.g. classical swine fever (CSF), foot–and–mouth disease (FMD), infectious bursal disease (IBD), bovine virus diarrhea (BVD), canine distemper, swine influenza (SI), chicken infectious anemia (CIA), avian influenza (AI), bluetongue disease, peste–des–petits ruminants (PPR), Newcastle disease (ND), sheep and goat pox, infectious bovine rhinotracheitis (IBR), Marek's disease (MD), pseudorabies, porcine reproductive and respiratory syndrome (PRRS), etc. have posed major challenges to field veterinarians. Need of Rapid Diagnostics: Traditional methods for viral detection include isolation, in vitro culture, immunoassays, and electron microscopy. These methods are expensive, require trained staff and time–consuming. These assays can not be performed on-site. In between the period of sample collection and result interpretation, in the meantime, there is risk of alleviating disease conditions, spread of infectious disease to more population and even death due to absence of appropriate disease management. Many of the major disease causing agent described above can only be handled in higher biosafety standards.
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In this scenario, rapid, on-site diagnosis of a pathogen will provide prompt measures to stop outbreak progression. One of such approach is a Biosensor device. A biosensor is a compact analytical device with a ligand– specific biorecognition element, e.g. antibody, enzyme, receptor, nucleic acid, aptamers, peptide/protein, lectin, cells, tissue or whole organisms, immobilized on a sensor surface integrated directly or indirectly with a signal conversion unit called transducer. Biosensors are broadly classified based on the biorecognition element and the transducers. Antibody–based biosensors employ antibody as biorecognition elements. Antibodies as Bio-recognition Element: Antibodies are Y–shaped proteins (immunoglobulins) that are, produced by plasma cells as host's immune response to counter antigen. Antibodies bind the target antigen and destroy it in order to protect the host. A typical antibody (immunoglobulin G/ IgG) is composed of a fragment antigen binding (Fab) region and a fragment crystallizable (Fc) region, made up of four polypeptide chains: two heavy chains (50 kDa) and two light chains (25 kDa). Antibodies used as probes can be polyclonal, monoclonal or recombinant. Transducer: A transducer converts the biorecognition phenomenon into a measurable signal. Many different types of transducers are present and they are classified as; 1. Electrochemical: production or consumption of the ions or electrons 2. Potentiometric: measure difference in potential 3. Impedimetric : measures the change in electrical resistance of the solution 4. Optical: measure changes in intensity of light 5. Mass–based: detect change in mass occurs following the interaction between the biorecognition element and the target analyte . Antibody immobilisation: An important factor that influences biosensing is the immobilization of the antibodies to the sensor surface. Various types of sensor surfaces like gold, silver, glass, platinum, silica are used. Ideal sensor surface should be stable, should possess excellent electrical and thermal conductivity and less signal–to–noise ratio due to matrix effects. Advances in Nanotechnology have produced high performance matrices such as carbon nanotubes, fabricated nanoparticles, self–assembled monolayers (SAMs) and quantum dots. These provide new generation sensor platforms compatible with the required sensor needs. The antibodies can Sr. No. Virus Antibody Type Transducer Type Detection Time 1 Rabies virus mAb Reflectance 30 min be coupled to the 2 Foot and Mouth Disease Virus mAb Optical Not mentioned sensor surface by 3 Porcine rotavirus pAb Reflectance 30 min various methods; 4 H1N1 pAb Surface plasmon resonance >20 minutes 1.Passive absorption 5 BVDV mAb Conductometric 8 min 2.Covalent coupling 6 H5N1 mAb Impedimetric >2 hours 3.Matrix entrapment 7 Infectious Bursal Disease mAb SPR Not specified 4.Encapsulation 8 Duck hepatitis virus serotype1 pAb Ellipsometry 30 min 5.Affinity tags Biosensors available for Veterinary Pathogens:
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Future prospects: Antibody–based sensors exhibit high sensitivity, high affinity and avidity with its respective antigen and thus, serve as good candidates for use with unprocessed samples. Most of the reported antibody–based biosensors use polyclonal or monoclonal antibodies. The availability of chicken antibodies and different recombinant antibody formats along with improved immobilization chemistries have helped overcome stability issues of test. Advances in nanotechnology and microfluidics led to development of point–of–care and lab–on–a–chip diagnostics. This has revolutionized the field of bio sensing and thus, there is increased interest in biosensors. Consequently, there is expansion in the biosensor market which is expected to be worth 27.06 Billion USD by 2022 (http://www.marketsandmarkets.com/PressReleases/biosensors.asp). However, the detection of veterinary pathogens is still to harness this technology on large scale to its benefit with bio sensing. Viral veterinary diagnostics can benefit from biosensors as they allow rapid, robust simple alternatives to conventional viral detection techniques. In addition, biosensors allow “on–site” or “pen-side” testing, and can be performed and interpreted, within a matter of minutes, by farmers or veterinarians. This is a highly desirable compared to collection and shipment of samples followed by time taken to get results. Consequently, biosensors can allow veterinarians to provide specific and timely treatment to animals thus reducing the morbidity and mortality. Additionally, it prevents the spread of contagious pathogens to another animals and humans (in case of zoonotic pathogens).
Priyanka .N., Pooja.C.R and Pushpa.G.Malagittimath Dept. of Veterinary Extension, Veterinary college. Shivamogga. Email I.D: pushpamalagittimath@gmail.com Hypogalactia may be defined as abrupt reduction in normal amount of milk production. It is not a disease but it is a condition/clinical sign. Causes of Hypogalactia: 1. Physiological causes & its pathogenesis: Udder edema: The condition results due to excess accumulation of fluid between secretary cells of the udder in high yielders after calving. Commonly occurs in heifers. Pathogenesis: Udder edema increases activity of the mammary gland leading to increase in blood supply to alveolar tissue of udder. And this increases capillary blood pressure& decreased plasma colloidal pressure leading to increase permeability of vascular endothelium causing movement of fluid to interstitial Udder edema space. It often extends forward under skin as fluid keeps on accumulating. Due to increase in pressure on the udder, milk let down is difficult. Hence decrease in milk yield. Estrus: Pathogenesis: Hormonal imbalance (increased estrogen level) & increased muscular activity, decreased feed intake. Temporarily seizes the prolactin secretion leading to decrease in milk secretion. Pregnancy: No effect on milk production in initial stages of pregnancy. It is due to less progesterone activity.
Pashubandha 2017 2014
Volume No :06 : 3 Issue Issue: 01 : 05
Pathogenesis: In advance stages of pregnancy there is increase in progesterone secretion this seizes prolactin and oxytocin secretion temporarily leading to decreased milk yield. 2. Pathological causes and its pathogenesis Mastitis Pathogenesis: Entry of causative agent through teat canal/systemic route. Invasive stage: Agent enters teat canal & mammary tissue cause inflammation in turn granulomatous tissue. Infective stage: Bacteria multiply cause damage to udder tissue. Septicemia & toxemia increase in temperature. Udder becomes hard painful/fibrotic/gangrenous. Decrease in milk production and let down. Trauma/Laceration Pathogenesis: Physical injury to the udder/wound. This trauma damages to the vascular system of udder leading to inflammation of udder (alveolar) tissue. Leads to hemogalactia/predispose to mastitis leads to decrease in milk production. Hypoplasia of udder Teat trauma Pathogenesis: Hormonal imbalance, deficiencies, chronic disease conditions leading to hypoplasia of alveolar tissue & duct system of mammary gland hence decrease in milk production. Metabolic disorders: Usually animal will be in negative energy balance in turn decreases the production (calf , milk ,beef) . 3. Environmental causes & pathogenesis Heat stress: Pathogenesis: Milking procedures and frequency: Knuckling procedure of milking may damages the teat cistern in turn results in decreased milk yield. Milking once /skipping the milking leading to increase in milk in udder, increases pressure. Decreased in milk production in alveolar tissue. Decrease in milk yield and letdown. DIAGNOSIS OF HYPOGALACIA: Udder edema: History of calving. Uniform swelling of udder & distended teats. Estrus: Signs of estrus. Pregnancy: By per rectal examination/ultrasonography. Mastitis: Physical examination of udder. Physical examination of milk. CMT test. ABST Trauma/laceration: History of trauma/laceration. Physical examination of udder. Swelling of part/ whole udder, erythematic, pain on palpation lacerations. Metabolic disorders: History of calving /advance Pregnancy.Symptoms: according to their deficiencies like ketosis fruity odor of breath. Estimation of biochemical profile like Ca, P, Mg & Serum glucose, enzymes. Heat stress: Seasonal, during summer. Increase in body temperature >39ĚŠ C. Open mouth breathing, salivation, dehydrated. TREATMENT FOR HYPOGALACIA AND ITS MANAGEMENT Udder edema: Treatment: Diuretics: Furosemide 1-2mg/kg bwt IM/IV 3 days. Carticosteroids:Dexamethasone 10-30 mg TD IM 3 days
Pashubandha 2017 2014
Volume No :06 : 3 Issue Issue: 01 : 05
Management: Frequent milking, once in 4 hr. Cold water splashing. Mastitis Antibacterial: Amoxicillin +clavulanic acid 8.75mg/kg bwt IM IV bid 5 days (Venclox, Glenmox –CL). Cefoperazone+salbactum 5-10 mg /kg bwt IM IV 5 days (Cephatin, Pathocef). Ceftriaxone +tazobactum 5 -10 mg /kg bwt IM IV 5 days (Intacef Tazo, Wocef - XP). Anti-inflammatory: Meloxicam 0.2-0.3 mg/kg bwt IM/IV sid 3-5days. Tolfenamic acid 2-4 mg/kg bwt IM/ IV sid 3-5 days. Intramammary infusions: Mastiwok oint (cefaperazone sodium 250mg). Pedistrin -SH (Procine penicillin 1 lac IU, streptomycin sulphate 100mg ,sulfamerazine 500mg,hydrocortisone acetate 29mg). Topical preparations: Mastilep gel 125g. Mastinil spray 500ml Heat stress Treatment: NSAIDS and fluid therapy if necessary. Management: Give cold water to drink. Cold water splashing. Put wet gunny bags on animal. Galactogogues: Dugdhdan: 4 tab bid oral. Lactomore powder 20 g daily. Other preparation includes OstoVet & Vimeral.
The National Bureau of Animal Genetic Resources (NBAGR) has conferred rare and singular species genetic recognition tag to the threatened breed of Kendrapara sheep found in Odisha. Kendrapara sheep is found only in Odisha. Locally it is called kuji mendha. The rare genetic traits lead to the multiple birth syndrome in them.Kendrapada sheep are primarily used for production of mutton. They are well adapted to high ambient temperature, high humidity and heavy rains.
monthly e-Bulletin Published and circulated by Veterinary College, Hebbal, Bengaluru. Editor: Dean, Veterinary College, Hebbal, Bengaluru Dr. H. N. NARASIMHA MURTHY (Ex-Officio)
Associate Editior: Head, Dept. of Vety.& Animal Husbandry Extension Education Dr. K. Satyanarayan (Ex-Officio)
Contact : Dept of Veterinary and Animal Husbandry Extension Education Veterinary College, Hebbal Bangalore email: pashubandhavch@gmail.com Blog: pashubandhavch.blogspot.in
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Pashubandha 2017 2014
PELVIC
Volume No :06 : 3 Issue Issue: 01 : 05