Journal of Research in Biology Volume 3 Issue 4 by Journal of Research in Biology

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List of Editors of Editors in the Journal of Research in Biology Managing and Executive Editor: Abiya Chelliah [Molecular Biology] Publisher, Journal of Research in Biology. Editorial Board Members: Ciccarese [Molecular Biology] Universita di Bari, Italy. Sathishkumar [Plant Biotechnologist] Bharathiar University. SUGANTHY [Entomologist] TNAU, Coimbatore. Elanchezhyan [Agriculture, Entomology] TNAU, Tirunelveli. Syed Mohsen Hosseini [Forestry & Ecology] Tarbiat Modares University (TMU), Iran. Dr. Ramesh. C. K [Plant Tissue Culture] Sahyadri Science College, Karnataka. Kamal Prasad Acharya [Conservation Biology] Norwegian University of Science and Technology (NTNU), Norway. Dr. Ajay Singh [Zoology] Gorakhpur University, Gorakhpur Dr. T. P. Mall [Ethnobotany and Plant pathoilogy] Kisan PG College, BAHRAICH Ramesh Chandra [Hydrobiology, Zoology] S.S.(P.G.)College, Shahjahanpur, India. Adarsh Pandey [Mycology and Plant Pathology] SS P.G.College, Shahjahanpur, India Hanan El-Sayed Mohamed Abd El-All Osman [Plant Ecology] Al-Azhar university, Egypt Ganga suresh [Microbiology] Sri Ram Nallamani Yadava College of Arts & Sciences, Tenkasi, India. T.P. Mall [Ethnobotany, Plant pathology] Kisan PG College,BAHRAICH, India. Mirza Hasanuzzaman [Agronomy, Weeds, Plant] Sher-e-Bangla Agricultural University, Bangladesh Mukesh Kumar Chaubey [Immunology, Zoology] Mahatma Gandhi Post Graduate College, Gorakhpur, India. N.K. Patel [Plant physiology & Ethno Botany] Sheth M.N.Science College, Patan, India. Kumudben Babulal Patel [Bird, Ecology] Gujarat, India.

Dr. Afreenish Hassan [Microbiology] Department of Pathology, Army Medical College, Rawalpindi, Pakistan. Gurjit Singh [Soil Science] Krishi Vigyan Kendra, Amritsar, Punjab, India. Dr. Marcela Pagano [Mycology] Universidade Federal de São João del-Rei, Brazil. Dr.Amit Baran Sharangi [Horticulture] BCKV (Agri University), West Bengal, INDIA. Dr. Bhargava [Melittopalynology] School of Chemical & Biotechnology, Sastra University, Tamilnadu, INDIA. Dr. Sri Lakshmi Sunitha Merla [Plant Biotechnology] Jawaharlal Technological University, Hyderabad. Dr. Mrs. Kaiser Jamil [Biotechnology] Bhagwan Mahavir Medical Research Centre, Hyderabad, India. Ahmed Mohammed El Naim [Agronomy] University of Kordofan, Elobeid-SUDAN. Dr. Zohair Rahemo [Parasitology] University of Mosul, Mosul,Iraq. Dr. Birendra Kumar [Breeding and Genetic improvement] Central Institute of Medicinal and Aromatic Plants, Lucknow, India. Dr. Sanjay M. Dave [Ornithology and Ecology] Hem. North Gujarat University, Patan. Dr. Nand Lal [Micropropagation Technology Development] C.S.J.M. University, India. Fábio M. da Costa [Biotechnology: Integrated pest control, genetics] Federal University of Rondônia, Brazil. Marcel Avramiuc [Biologist] Stefan cel Mare University of Suceava, Romania. Dr. Meera Srivastava [Hematology , Entomology] Govt. Dungar College, Bikaner. P. Gurusaravanan [Plant Biology ,Plant Biotechnology and Plant Science] School of Life Sciences, Bharathidasan University, India. Dr. Mrs Kavita Sharma [Botany] Arts and commerce girl’s college Raipur (C.G.), India. Suwattana Pruksasri [Enzyme technology, Biochemical Engineering] Silpakorn University, Thailand. Dr.Vishwas Balasaheb Sakhare [Reservoir Fisheries] Yogeshwari Mahavidyalaya, Ambajogai, India.

CHANDRAMOHAN [Biochemist] College of Applied Medical Sciences, King Saud University.

Dr. Pankaj Sah [Environmental Science, Plant Ecology] Higher College of Technology (HCT), Al-Khuwair.

B.C. Behera [Natural product and their Bioprospecting] Agharkar Research Institute, Pune, INDIA.

Dr. Erkan Kalipci [Environmental Engineering] Selcuk University, Turkey.

Kuvalekar Aniket Arun [Biotechnology] Lecturer, Pune.

Dr Gajendra Pandurang Jagtap [Plant Pathology] College of Agriculture, India.

Mohd. Kamil Usmani [Entomology, Insect taxonomy] Aligarh Muslim university, Aligarh, india.

Dr. Arun M. Chilke [Biochemistry, Enzymology, Histochemistry] Shree Shivaji Arts, Commerce & Science College, India.

Dr. Lachhman Das Singla [Veterinary Parasitology] Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, India.

Dr. AC. Tangavelou [Biodiversity, Plant Taxonomy] Bio-Science Research Foundation, India.

Vaclav Vetvicka [Immunomodulators and Breast Cancer] University of Louisville, Kentucky.

Nasroallah Moradi Kor [Animal Science] Razi University of Agricultural Sciences and Natural Resources, Iran

José F. González-Maya [Conservation Biology] Laboratorio de ecología y conservación de fauna Silvestre, Instituto de Ecología, UNAM, México.

T. Badal Singh [plant tissue culture] Panjab University, India

Dr. Kalyan Chakraborti [Agriculture, Pomology, horticulture] AICRP on Sub-Tropical Fruits, Bidhan Chandra Krishi Viswavidyalaya, Kalyani, Nadia, West Bengal, India. Dr. Monanjali Bandyopadhyay [Farmlore, Traditional and indigenous practices, Ethno botany] V. C., Vidyasagar University, Midnapore. M.Sugumaran [Phytochemistry] Adhiparasakthi College of Pharmacy, Melmaruvathur, Kancheepuram District. Prashanth N S [Public health, Medicine] Institute of Public Health, Bangalore. Tariq Aftab Department of Botany, Aligarh Muslim University, Aligarh, India. Manzoor Ahmad Shah Department of Botany, University of Kashmir, Srinagar, India. Syampungani Stephen School of Natural Resources, Copperbelt University, Kitwe, Zambia. Iheanyi Omezuruike OKONKO Department of Biochemistry & Microbiology, Lead City University, Ibadan, Nigeria. Sharangouda Patil Toxicology Laboratory, Bioenergetics & Environmental Sciences Division, National Institue of Animal Nutrition and Physiology (NIANP, ICAR), Adugodi, Bangalore. Jayapal Nandyal, Kurnool, Andrapradesh, India. T.S. Pathan [Aquatic toxicology and Fish biology] Department of Zoology, Kalikadevi Senior College, Shirur, India. Aparna Sarkar [Physiology and biochemistry] Amity Institute of Physiotherapy, Amity campus, Noida, INDIA. Dr. Amit Bandyopadhyay [Sports & Exercise Physiology] Department of Physiology, University of Calcutta, Kolkata, INDIA . Maruthi [Plant Biotechnology] Dept of Biotechnology, SDM College (Autonomous), Ujire Dakshina Kannada, India. Veeranna [Biotechnology] Dept of Biotechnology, SDM College (Autonomous), Ujire Dakshina Kannada, India. RAVI [Biotechnology & Bioinformatics] Department of Botany, Government Arts College, Coimbatore, India. Sadanand Mallappa Yamakanamardi [Zoology] Department of Zoology, University of Mysore, Mysore, India. Anoop Das [Ornithologist] Research Department of Zoology, MES Mampad College, Kerala, India.

Dr. Satish Ambadas Bhalerao [Environmental Botany] Wilson College, Mumbai Rafael Gomez Kosky [Plant Biotechnology] Instituto de Biotecnología de las Plantas, Universidad Central de Las Villas Eudriano Costa [Aquatic Bioecology] IOUSP - Instituto Oceanográfico da Universidade de São Paulo, Brasil M. Bubesh Guptha [Wildlife Biologist] Wildlife Management Circle (WLMC), India Rajib Roychowdhury [Plant science] Centre for biotechnology visva-bharati, India. Dr. S.M.Gopinath [Environmental Biotechnology] Acharya Institute of Technology, Bangalore. Dr. U.S. Mahadeva Rao [Bio Chemistry] Universiti Sultan Zainal Abidin, Malaysia. Hérida Regina Nunes Salgado [Pharmacist] Unesp - Universidade Estadual Paulista, Brazil Mandava Venkata Basaveswara Rao [Chemistry] Krishna University, India. Dr. Mostafa Mohamed Rady [Agricultural Sciences] Fayoum University, Egypt. Dr. Hazim Jabbar Shah Ali [Poultry Science] College of Agriculture, University of Baghdad , Iraq. Danial Kahrizi [Plant Biotechnology, Plant Breeding,Genetics] Agronomy and Plant Breeding Dept., Razi University, Iran Dr. Houhun LI [Systematics of Microlepidoptera, Zoogeography, Coevolution, Forest protection] College of Life Sciences, Nankai University, China. María de la Concepción García Aguilar [Biology] Center for Scientific Research and Higher Education of Ensenada, B. C., Mexico Fernando Reboredo [Archaeobotany, Forestry, Ecophysiology] New University of Lisbon, Caparica, Portugal Dr. Pritam Chattopadhyay [Agricultural Biotech, Food Biotech, Plant Biotech] Visva-Bharati (a Central University), India

Table of Contents (Volume 3 - Issue 4) Serial No

Accession No

RA0344

Title of the article

Influence of the growing area on oil palm (Elaeis guineensis) inflorescences insects population.

Page No

940-946

Koua Kouakou HervĂŠ, Akpesse Apka Alexandre Moise, TUO Yalamoussa, and Hala Nâ&#x20AC;&#x2122;klo. 2

RA0349

A chromosomal analysis of seven Cameroonian Acrididae species (Orthoptera: Acridinae, Oedipodinae and Spathosterninae) based on published data.

947-953

Seino Richard Akwanjoh and Dongmo Tonleu Ingrid. 3

RA0324

Impact of the residue of Deltamethrin and Endosulfan pesticides on biochemical toxicity and some neurotransmitter contents in different brain areas of male Albino mice.

954-966

Somaya M. Ismail, Azza A. Said and Samira M. El-Sayad. 4

RA0342

Prevalence and the effect of plant extracts on community associated methicillin resistant Staphylococcus aureus in Owerri, Imo State, Nigeria.

967-976

Amadi ES, Oguoma OI, Ibekwe VI, Abanobi SE, Chikwendu CI and Egbadon OE. 5

RA0346

Odonata diversity (Insecta: Arthropoda) in rice and vegetable fields in a north-eastern district of Tamil Nadu, India.

977-983

Veeramuthu Anbalagan, Michael Gabriel Paulraj and Savarimuthu Ignacimuthu. 6

RA0343

Hepatic enzyme markers and proteins in serum and some selected tissues in Clarias gariepinus from swamp around Kokori-Erhoike oil field, Nigeria. Osioma E, Akanji MA and Arise RO.

984-992

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Influence of the growing area on oil palm (Elaeis guineensis) inflorescences insects population Authors: Koua Kouakou Hervé1, Akpesse Apka Alexandre Moise2, Tuo Yalamoussa3, and Hala N’klo4. Institution: 1. Félix Houphouet-Boigny University of Cocody (Abidjan, Côte d’Ivoire). 22 BP: 1611 Abidjan 22. 2. Félix Houphouet-Boigny University of Cocody (Abidjan, Côte d’Ivoire). BP: 582 Abidjan 22. 3. Félix Houphouet-Boigny University of Cocody (Abidjan, Côte d’Ivoire). BP: 582 Abidjan 22.

ABSTRACT:

Oil palm tree grows naturally on low ground and on plain. Seed production varies from one area to another on the same oil palm plantation. Pollination of oil palm is essentially entomophilous; it appeared useful to assess the influence of the growing area on the fluctuation of pollinating insects’ population. Samplings were performed each month on male and female inflorescences during two years on plots in lowland and plain. The insects showed no qualitative change from one area to another. Sixteen species of insects were observed on the male inflorescences against 10 species on female inflorescences. The inflorescences showed variation in the number of insects based on the growing area and the stage of flowering.

Keywords: culture area; pollinating insects; Lamé; Côte d’Ivoire.

4. National Center of Agronomic Research (CNRA, Côte d’Ivoire) BP: 1740 Abidjan 01.

Corresponding author: Koua Kouakou Hervé.

Article Citation: Koua Kouakou Hervé, Akpesse Apka Alexandre Moise, TUO Yalamoussa, and Hala N’klo. Influence of the growing area on oil palm (Elaeis guineensis) inflorescences insects population. Journal of Research in Biology (2013) 3(4): 940-946

Web Address:

Dates:

http://jresearchbiology.com/ documents/RA0344.pdf.

Received: 25 Mar 2013

Accepted: 02 May 2013

Published: 16 May 2013

This article is governed by the Creative Commons Attribution License (http://creativecommons.org/ licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.

Journal of Research in Biology An International Scientific Research Journal

940-946 | JRB | 2013 | Vol 3 | No 4

www.jresearchbiology.com

Koua et al., 2013 insects in two agro ecosystems: lowland and plain.

INTRODUCTION Many problems

are opposed

to a

good

production of oil palm. These include pests, pathogenic

MATERIAL AND METHODS

fungi and especially the continuing decline of pollinating

Study site

insects. For several years many problems of fruit set

Our study site was the experimental station of La

were observed in some regions of cultivation of oil palm

Mé, located at 5°26, N, 3°50, W. The station is located to

tree causing a gradual decline in seed production (Mariau

about thirty kilometers north-east of Abidjan (Côte

et al., 1991). The observation was made that the most

d’Ivoire). This area is characterized by an ombrophilous

affected areas, spread over large areas of lowland.

forest (Traoré and Mangara, 2009).

There are many evidences that the pollinator

The study area has an equatorial climate

insects effectively contributes to the reproduction of

characterized by two distinct rainy seasons (March to

many cultivated plant species. Regarding the oil palm

July and November). These two seasons are alternated by

tree, the discovery of the role of insects in pollination

two dry seasons: December to February and from August

was made by Chevalier in 1910. The works of Syed

to October (Pene and Assa, 2003). The monthly mean

(1979) and Syed et al. (1982) have confirmed this

temperature was about 27°C. The monthly average of the

finding.

essentially

highest temperature was recorded in March and the

entomophilous (Corrado, 1985). Without pollination,

lowest in August with respectively 28.55 and 25.5°C.

fruit set by wind is extremely low (Mariau et al., 1991).

The average annual rainfall was about 1500 mm. The

Pollinating insects are thus an undeniable role. In Côte

average annual sunshine duration was about 1790 h; the

d'Ivoire (West Africa), five species of Elaeidobius

average monthly relative humidity was about 81%.

(E.

Insects of male inflorescences

Pollination

kamerunicus,

oil

palm

plagiatus,

subvitatus,

E. bilineatus, E. singularis), two species of Microporum

Three operations were performed to assess the

(M. congolense and M. dispar), two species of

male inflorescences of oil palm insect fauna: location,

Proseostus (P. minor, P. sculptilis), Atheta burgeoni,

sampling and identification of insects (Fataye, 1984).

Gabrius sp, Litargus sp, Thrips sp and Anthocoris sp

Location

have been described by Desmier De Chenon, (1981) and

Each month a location was carried out in order to

Hala et al., (2012) as pollinators of oil palm. It therefore

count four inflorescences in the process of flowering.

seemed appropriate to follow the dynamics of these

This location has identified 192 male inflorescences at a

insects on two different ecological zones: lowland and

rate of eight per month during two years.

plain. The research question that we asked is whether the

Sampling

growing area of oil palm has an influence on populations

As soon as the third florets of each inflorescence

of pollinating insects. Many factors can explain the

were listed, with pair of secateurs four spikelets per

fluctuations of insect populations. The latest studies on

inflorescence were collected. This collection was done in

this subject have established outside the bioclimatic

the beginning of anthesis (BA). The second (full

factors which have a clear implication, that the use of

anthesis: FA) and third (end anthesis: EA) took place

insecticides in the fight against pests do not spare

respectively after the first three days and two days after

beneficial insects that are pollinators (Tuo et al., 2011).

the second. Each batch of four spikelets collected was

This preliminary study established a quantitative and qualitative comparison of oil palm inflorescences 941

placed in a bag and then the insects collected were neutralized

with

insecticide

bomb.

Before

Journal of Research in Biology (2013) 3(4): 940-946

Koua et al., 2013 identification, insects of each batch of spikelet were

classify the means into homogeneous groups

collected in pillboxes containing alcohol 70%. Identification

RESULTS AND DISCUSSION

Using the collection of the insect fauna of oil palm inflorescences of the National Agricultural

Variation in the number of insects on male inflorescences

Research Centre of La ME and a binocular microscope,

On male inflorescences of oil palm tree, the

insects of every month and each area were identified at

insects mostly belong to Elaeidobius (E) genus. Five

the species level.

species were observed: E. kamerunicus, E. plagiatus,

Insects of female inflorescences

This study was conducted according to the

subvittatus,

singularis and E.

bilineatus.

Microporum (M) genus was represented by the species

methods of N'goran (1982) and Fataye (1984).

M. dispar and M. congolense. Prosoestus genus was

Location

present with two species P. sculptilis and P. minor.

Two non-flowering inflorescences per plot were

Species, Atheta burgeoni, Lithargus sp., Anthocoride sp.,

identified and followed by month. The inflorescences

Thrips sp, Gabrius sp. and bees (Nomia sp and

were cleared of husks and bulky leaves with machetes

Apis mellifera) were also observed.

and

At the beginning of anthesis (BA)

knives

three

days

before

flowering.

Each

inflorescence thus revealed was covered with a muslin

The number of insects collected from the plot of

cage and attached to the floral stem with a rubber. The

lowland (61%) is higher than that collected on the plain

bagged inflorescences were controlled each afternoon to

(31%). At the species level, it was observed that

follow the evolution of the inflorescence.

E. singularis, E. bilineatus, P. sculptilis, M congolense

Sampling

and Anthocoris sp showed a significant difference

All the insects that were attracted are placed on

depending on the growing area with a higher effective in

the cage once flowering commences. Using a vacuum

lowland areas. The other species showed no preference

cleaner, these insects were captured every hour for ten

for one area (Figure 1A).

minutes. This operation was performed at 6 AM to 6 PM

At full anthesis (FA)

during the two days of the anthesis length. The collected

The total number of insects collected was 42% in

insects were immediately stored in pillboxes containing

the plain region and 58% in lowland areas. For the

70% alcohol. At the end of the day, insects collected

species, E. plagiatus, E. kamerunicus, M. congolense,

were sent to the laboratory. At each study site, sampling

M. dispar and A. Burgeoni, a significant difference was

was conducted on 48 inflorescences.

found between their respective populations based on the

Identification

growing area. Only A. burgeoni presented a higher

Insects collected were identified using the same protocol as previously.

effective in the lowland areas. The other species were much more present in plain areas. Besides these species, no differences were recorded between the number of

STATISTICAL ANALYSIS Data processing was performed using Statistica software version 7.1. An analysis of variance (ANOVA)

insects collected in lowland areas and those collected on the plain (Figure 1B). At the end of anthesis (EA)

revealed significant differences between the data. The

The total number of insects differ from one area

test of Student-Newman-Keuls at 5% was used to

to another. It was 75% in lowland areas against 25% on

Journal of Research in Biology (2013) 3(4): 940-946

942

Koua et al., 2013 the plain. With regard to species, only the number of E. singularis, M. dispar, Lithargus sp and A. burgeoni depends on the area. These species excepted Lithargus sp were more abundant in the lowland than on the plain (Figure 1C). Variation in the number of insects on female inflorescences The species observed on female inflorescences A: Beginning of anthesis

were: E. kamerunicus, E. plagiatus, E. subvittatus, E. bilineatus, E. singularis, M. congolense, M. dispar, P. minor, P. sculptilis and Atheta burgeoni. First day of anthesis The total population of insects was significantly higher on the plain (78%) than in the lowland area (22%). At the species level, only the species M. dispar, M.

congolense,

plagiatus,

kamerunicus,

E. subvittatus, E. singularis, A. burgeoni and P. minor,

B: Full anthesis

were affected by the growing area. A. burgeoni attended more inflorescences of the lowland area. Other species were more present on the plain than in the lowlands (Figure 2A). Second day of anthesis Insects were relatively influenced by the growing area. Indeed, 78% of the insects were collected on inflorescences of the plain region against 28% in the lowlands. Regarding species, E. subvittatus, A. Burgeoni,

C: End of anthesis

P. minor and P. sculptilis, had no preference for the

Lowland

growing area. The species E. kamerunicus, E. plagiatus,

Histograms with the same letter significantly different at the 5% level

E. bilineatus, E. singularis, M. dispar and M. congolense

Plain are

not

were receptive to the area of culture (Figure 2B).

Figure 1: Influence of the growing area on the Among the species, only E. kamerunicus, number of insects present on male inflorescences E.su: Elaeidobius subvittatus; E.p: Elaeidobius E. plagiatus, E. singularis and M. dispar were affected plagiatus; E.s: Elaeidobius singularis; E.b: Elaeidobius by the growing area both the first and second day of balineatus; E.k: Elaiedobius kamerinucus; P.m: Prosoestus minor; P.s: Prosoestus sculptilis; anthesis. M.c: Microporum congolense; M.d: Microporum The insects of oil palm inflorescences showed no dispar; L.sp: Lithargus sp.; Ant.sp.: Anthocoris sp.; A.bur: Atheta burgeoni: T.sp.: Thrips sp.; G.sp.: qualitative change from one area to another. These Gabrius sp. species were always present on the oil palm tree and

Sixteen insect species were observed on the male

colonize the inflorescences of this plant regardless of the

inflorescences against only 10 species on female

study area.

inflorescences. The six species that were absent on the

943

Journal of Research in Biology (2013) 3(4): 940-946

Koua et al., 2013 female inflorescences (Lithargus sp., Anthocoride sp., Thrips sp., Gabrius sp., and bees (Nomia sp. and Apis mellifera) would not intervene mainly in oil palm pollination. According to Mariau et al., 1991, four species provide the largest share of pollination of oil palm tree: E. kamerinucus, E. plagiatus, E. subvittatus and E. singularis. The male inflorescences showed a variation in the numbers of insects based on the growing area and the

A: First day of anthesis

stage of flowering. In general, the lowland areas showed significantly more insects than upland areas (61% against 31% at the beginning of anthesis, 58% against 42% at full anthesis and finally 75% against 25% at the end of anthesis). Insects observed in the male inflorescences live on them usually. For example, the male flowers are the breeding sites of insects of the genus Elaeidobius (Beaudoin-Ollivier et al., 2012). The differences could be explained by changes in environmental factors. At the A: Second day of anthesis

beginning of anthesis, flowers began to appear on the male inflorescences that induced an attractive factor because of the strong smell of anise emitted by the male

Lowland

Plain

Histograms with the same letter are not flowers. To this, were added the environmental factors significantly different at the 5% level. including relative humidity and temperature. It was noted Figure 2: Influence of the growing area on the that the number of insects on lowland inflorescences numbers of insects present on female inflorescences E.su: Elaeidobius subvittatus; E.p: Elaeidobius were two times higher than those of insects collected on plagiatus; E.s: Elaeidobius singularis; E.b: Elaeidobius plain region inflorescences. At the species level, if for balineatus; E.k: Elaeidobius kamerinucus; P.m: Prosoestus minor; P.s: Prosoestus sculptilis; M.c: E. bilineatus, P. sculptilis and M. congolense, numbers Microporum congolense; M.d: Microporum dispar; were highest in the lowland area than on the plain region. L.sp: Lithargus sp.; Ant.sp.: Anthocoris sp.; A.bur: Atheta burgeoni: T.sp.: Thrips sp.; G.sp.: Gabrius sp. These three species are unfortunately not effectively intervening in the pollination of oil palm tree. The other

M. congolense were significantly different from one area

insect species showed no significant difference at the

to another. Indeed, apart A. burgeoni, all these species

beginning of anthesis.

were more prevalent in upland areas. This can be

In full anthesis, the number of insects on male inflorescences has reached its maximum value. The

explained by bioclimatic factors which were more favorable to the activity of these insects.

attractive factor of flowers took over on bioclimatic

We observed three times more insects on

factors. Thus, it has almost the same number of insects

lowland inflorescences than on those on plain at the end

from one region to another. In terms of species observed

of anthesis. This can be explained by the fact that on

full

plain, the flowers were fading and dry faster than the

kamerunicus,

anthesis,

the A.

number Burgeoni,

of M.

plagiatus,

dispar

Journal of Research in Biology (2013) 3(4): 940-946

and

inflorescences in lowland areas (lower temperature and 944

Koua et al., 2013 higher relative humidity). Insects were removed quickly

Chevalier A. 1910. Documentation sur le palmier à

on inflorescences located in plain region. At the species

huile, végétaux utiles de l’Afrique Tropicale- VII - Paris,

level, only E. singularis, M. dispar, A. burgeoni and Lithargus sp. showed significantly different numbers

Corrado F. 1985. La conformation des régimes de

from

palmier à huile (Elaeis guineensis Jacq.) dans quelques

one

requirements

area of

to another. these

species

Only bio-ecological can

explain

this

plantations de Colombie. Oléagineux, 40 (4):173-187.

distribution. According to Hussein et al., 1990, the change in the population of pollinating insects in

Desmier De Chenon R. 1981. Entomophil pollination of

plantations of oil palm is largely due to the influence of

oil palm in West Africa. Preliminary research. In: The oil

intrinsic and extrinsic factors, in particular, biological

palm in agriculture in the eighties. Incorporated Society

and chemical factors.

of Planters ed., Malaysia, Vol. I, 239-291.

During the anthesis, the total number of insects is higher on female inflorescences taken from the plain

Fataye A. 1984. Rôle des principaux insectes dans la

region than in the lowland area. At the species level, only

pollinisation des palmiers à huile en Côte d’Ivoire.

P. sculptilis showed no preference zone during the two

Rapport de stage de fin de première année agronomique,

days during anthesis. The other

ENSA, Abidjan - Station palmier à huile IRHO-CIRAD

species except

A. burgeoni showed a preference for the plateau region.

de La Mé, CI, 26.

The determining factor is mainly the higher temperature which allows the presence and maximal activity of

Hala N, Tuo Y, Akpesse AAM, Koua HK and Tano

insects found on female inflorescences (Mariau et al.,

Y. 2012. Entomofauna of Oil Palm Tree Inflorescences

1991). Thus, the first day of anthesis as the second, the

at La Mé Experimental Station (Côte d’Ivoire).

numbers of insects were higher in these areas.

American Journal of Experimental Agriculture 2(3): 306319.

CONCLUSION The number of insects collected on the plain

Hussein MY, Lajis NH and Ali JH. 1990. Biological

region is significantly different from that harvested the

and chemical factors associated with the successful

lowland area. The number of insect has been higher in

introduction of Elaeidobius kamerunicus faust, the oil

male inflorescences in lowland areas than on the plain.

palm pollinator in Malaisia. Acta Horticulturae, 288: 81-

This number was higher on the plain than in the lowland

87.

area. The numbers of insects are influenced by the culture area. This factor is to be taken into account in the

Mariau D, Houssou M, Lecoustre R and Ndigui B.

implementation and the entomological monitoring of oil

1991. Insectes pollinisateurs du palmier à huile et taux de

palm plantations.

nouaison en Afrique de l’ouest ; Oléagineux., Vol. 46 (2) : 43-51.

REFERENCES Beaudoin-Ollivier L, Frerot B, Coffi A, Flori A. 2012.

N’goran DF. 1982. Etude du trafic des insectes sur les

Duperie olfactive et pollinisation chez le palmier.

fleurs femelles des palmiers à huile ; importance des

Journée Filière Palmier à Huile – juillet Cirad, Paris

populations sur les fleurs mâles. Rapport de stage de fin de première année agronomique, ENSA, Abidjan-

945

Journal of Research in Biology (2013) 3(4): 940-946

Koua et al., 2013 Station palmier à huile IRHO-CIRAD de La Mé, CI.13p. Syed RA. 1979. Studies on oil palm pollination by insects. Bull. Ent. Res, 69(2): 213-224. Syed RA, Law IH and Corley RH. 1982. VInsect pollination of oil palm: introducing, establishment and pollinating efficiency of E. kamerunicus FRAUSTY in Malaysia. Incorporated Society of Planters ed. Vol. 58, 547-560. Traoré

and

Phyto-écologique

Mangara, des

2009.

Adventices

dans

Etude les

Agroécosystèmes élaeicoles de la Mé et de Dabou. Eur. J. Sci. Res., 65, 519-533. Tuo Y, Akpesse AAM, Hala N and Koua HK. 2011. Impact of terrestrial spraying of thiocyclam hydrogen oxalate on oil palm pollinating insects. Journal of Agriculture and Biological Sciences Vol. 2(7) pp.208 213.

Submit your articles online at www.jresearchbiology.com Advantages

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Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

A chromosomal analysis of seven Cameroonian Acrididae species (Orthoptera: Acridinae, Oedipodinae and Spathosterninae) based on published data Authors: ABSTRACT: Seino Richard Akwanjoh1,2 and Dongmo Tonleu Ingrid1. So far, the karyotypes of seven Acrididae species from Cameroon have been reported. These species included: Acrida turrita, Chirista compta, Coryphosima stenoptera producta, Oxycatantops spissus (Acridinae), Paracinema luculenta, Morphacris fasciata (Oedipodinae) and Spathosternum pygmaeum (Spathosterninae). Institution: Karyotype and meiosis relationships among these species were analysed from 1. Laboratory of Applied published data. The species had a common karyotype made up of 23 acrocentric Ecology (LABEA), chromosomes (males), the sex mechanism in all seven species was XXâ&#x2122;&#x20AC;-XOâ&#x2122;&#x201A; Department of Animal Biology, Faculty of Science, and meiosis was normal and chiasmate. The chromosomes in the species occurred in three size groups of long, medium and short. The number of chromosomes per size University of Dschang, group however varied among the species (A. turrita = 4LL:5MM:2SS; P.O. Box 353, Dschang, C. compta =4LL:4MM:3SS; C. stenoptera product=2LL:6MM:3SS; O. spissus Cameroon. =5LL:3MM:3SS; P. luculenta = 6LL:2MM:3SS; M. fasciata = 6LL:2MM:3SS; and 2. Department of Biological S. pygmaeum = 2LL:7MM:2SS). The X chromosome was long in the Oedipodinae, Science, Faculty of Science, medium in the Acridinae and short in the Spathosterninae. Total length of University of Bamenda, chromosomal material was in the series C. compta > O. spissus > P. luculenta > P.O. Box 39, Bamenda, S. pygmaeum > A. turrita > M. fasciata > C.s. producta. Cameroon. Corresponding author: Seino Richard Akwanjoh

Keywords: Acrididae, Acridinae, Oedipodinae, Spathosterninae, karyotype, relationships.

Email: raseino@yahoo.co.uk

Article Citation: Seino Richard Akwanjoh and Dongmo Tonleu Ingrid. A chromosomal analysis of seven Cameroonian Acrididae species (Orthoptera: Acridinae, Oedipodinae and Spathosterninae) based on published data. Journal of Research in Biology (2013) 3(4): 947-953

Web Address:

Dates:

http://jresearchbiology.com/ documents/RA0349.pdf.

Received: 15 Apr 2013

Accepted: 02 May 2013

Published: 20 May 2013

Journal of Research in Biology An International Scientific Research Journal

947-953 | JRB | 2013 | Vol 3 | No 4

www.jresearchbiology.com

Seino and Dongmo, 2013 Coryphosima stenoptera producta, Oxycatantops spissus

INTRODUCTION The use of Orthoptera material for karyotype

(Sub-family

Acridinae)

Paracinema

luculenta,

studies dates from the inception of cytogenetics. This is

Morphacris fasciata (Sub-family Oedipodinae) and

simply because Orthoptera material presents large

Spathosternum pygmaeum (Sub-family Spathosterninae).

chromosomes and few chromosomes per karyotype.

The species and the sources from which karyotypic

Chromosome size and number are of important

information on them was obtained for this analysis are

cytotaxonomic value (Turkoglu and Koca, 2002). The

shown in Table 1.

Orthoptera are also well known for their karyotypic

To analyse these karyotypes for similarities and

uniformity in chromosome number and morphology

differences, the karyotypes of the seven species were

(Ashwathanarayana and Ashwath, 2006; Chadha and

also arranged together (Figure. 1) and the morphometric

Mehta, 2011a).

characters for the seven species were arranged in a

It has been severally shown that analysis of

tabular form (Table 2).

karyotype differentiation between species yields better understanding of the evolutionary interrelationships and

RESULTS

divergence (Chadha and Mehta, 2011a; Sandhu and

Information

chromosome

number,

Chadha, 2012). A survey of investigations on karyotype

morphology, size, and length of X chromosome obtained

evolution in different groups of animals has revealed that

for the seven species is summarised in Table 2. A perusal

several karyotypes are dynamic and are subject to

of Table 2 revealed that among the seven species studied

change. Therefore, the stable karyotypes of the Acrididae

A. turrita, C. compta, C. stenoptera producta and

are subject to change.

O. spissus belonged to the sub-family Acridinae,

The cytogenetic diversity of Cameroonian

P. luculenta and M. faciata belonged to the subfamily

acridid grasshoppers has not been investigated. During

Oedipodinae and S. pygmaeum belonged to the

this study, published data on karyotypic characters were

subfamily Spathosterninae (Mestre and Chiffaud, 2009).

analysed to determine similarities and differences as well

Table 2 also revealed that the seven species investigated

had a common a diploid chromosome number of 2n=23

interrelationships

among

seven

Cameroonian

Acrididae species.

and the sex determining mechanism was XO in males. Figure 1 also revealed that the in the seven species investigated was acrocentric in morphology. The

MATERIALS AND METHODS The cytogenetics of only seven Cameroonian

chromosomes in all seven species occurred in three size

Acrididae species have so far been described. The

groups of long, medium and short. The number of

species

chromosome pairs per size group varied between species

include

Acrida turrita,

Chirista

compta,

Table 1: The species analysed, their subfamilies and references from which karyotypic information was obtained S/No 1 2 3 4 5 6 7 948

Species Acrida turrita Chirista compta Coryphosima stenoptera producta Oxycatantops spissus Paracinema luculenta Morphacris fasciata Spathosternum pygmaeum

Subfamily Acridinae

Oedipodinae Spathosterninae

Source of data Seino et al, 2008 Seino et al, 2010 Seino et al, 2010 Seino et al, 2010 Seino et al, 2012 Seino et al, 2012 Seino et al, 2012

Journal of Research in Biology (2013) 3(4): 947-953

Seino and Dongmo, 2013

Nature of X chromosome

and subfamilies (Table 2; Figure. 2). The Oedipodinae showed most similarity since both of them revealed 3SS)

6 long, 2 medium and 3 short chromosomes (6LL: 2MM: in

their

karyotypes.

The

lengths

the

C. compta > O. spissus > C.s. producta > M; fasciata >

1.7±0.00

5.5±0.24

7.7±0.59

6.6±0.00

5.6±0.56

7.3±0.52

5.0±0.08

Length (μm ) of X chromosome

X chromosome was in the series P. luculenta > A. turrita > S. pygmaeum. However, the X chromosome

All

160.7±0.91

129.9±0.63

164.7±2.26

165.4±0.61

123.6±7.60

A. turrita> M. fasciata > C.s. producta.

176.3±0.14

C. compta> O. spissus> P. luculenta> S. pygmaeum>

total length of chromosomal material was in the series

134.6±0.79

A M

Morphology of chromosomes Total chromosome length (μm)

and short in the Spathosterninae species (Figure. 2). The

DISCUSSION Every species has a unique karyotype which

Journal of Research in Biology (2013) 3(4): 947-953

characterised by a basic karyotype of 23 chromosomes in M= Metacentric, SM= Submetacentric, A= Acrocentric; L=long, M=Medium, S=Short

7 2 XX-XO 23 Spathosternina S. pygmaeum 7

3 2 6 XX-XO 23 Oedipodinae M. fasciata 6

3 2 6 XX-XO 23 Oedipodinae P. luculenta 5

3 3 5 XX-XO 23 Acridinae O. spissus 4

3 6 2 XX-XO 23 Acridinae C. S. producta 3

3 4 4 XX-XO 23 Acridinae C. compta 2

2 5 4 XX-XO 23 Acridinae A. turrita

Species

Sub- family S.N

Ranganath, 1997). Acridid grasshoppers are known to be

♀-♂

Sex determining mechanism

Long

Medium

Number of chromosome per size group

Short

provides an identity to the species (Channaveerappa and

Total number of chromosomes per cell in the male

Table 2: Morphometric characters of karyotypes of the seven species investigated

was medium in the Acridinae, long in the Oedipodinae

males. Due to this great cytogenetic uniformity Acridids are

considered

example

‘karyotypic

conservation’ (Aswathanarayana and Aswath, 2006). In the present study, seven Acridids have been investigated which belong to three different sub-families that

include

the

Acridinae,

Oedopodinae

and

Spathosterninae. The results of this study revealed that the seven Acrididae have a chromosome number of 23 and a sex determining mechanism which is XO/XX. Similar observations have been made for several other Acrididae species. With respect to chromosome number, chromosome

morphology

and

sex

determining

mechanism, Bugrov et al., (1994); Bugrov (1995), Bugrov et al., (1999) Bugrov and Sergeev (1997) observed

similar

results

for

Podisma

and

Eyprepocnemidinae (Acrididae) grasshoppers in Russia and Central Asia. Camacho and Cabrero (1983) also reported similar results for European species of Acrotylus (Oedopodinae). Mehta

(2011a)

Spathosternum

Yao

(2006)

reported

and similar

pransiniferum

Chadha

and

results

for

(Spathosterninae) 949

Seino and Dongmo, 2013

g Figure. 1: Mitotic Metaphase chromosomes in the seven species investigated. a) Acrida turrita, b) Chirista compta, c) Coryphosima stenoptera producta, d) Oxycatantops spissus, e) Paracinema luculenta, f) Mophacris fasciata, g) Spathosternum pygmaeum. Chromosomes are tapered towards one end and centromeres were deemed to be towards the tapered ends of the chromosomes. respectively from Asia and India. So the Acridid

Acrididae species (White, 1973; Sharma and Gautam,

grasshoppers

showing

2002; Mayya et al., 2004; Chadha and Mehta, 2011a).

cytogenetic uniformity regarding chromosome number,

Turkoglu and Koca (2002) reported the presence of

morphology and sex determining mechanism. The results

metacentric,

of this study confirmed that the basic Acrididae

chromosomes in the karyotypes of Oedipoda schochi and

karyotype is 23 acrocentric chromosomes and a sex

Acrotylus insbricus (Oedopodinae) from Turkey. The

determining

type.

aberrant chromosomes were the result of centric fissions.

Metacentric chromosomes through fusions were not

X - autosome fusion resulting in the Neo - XY sex

observed in the seven species here investigated even

mechanism have been reported in some acridid

though they have been reported in several other

grasshoppers (White, 1973). Bidau and Marti (2000)

950

different

mechanism

regions

the

are

XX/XO

submetacentric

and

acrocentric

Journal of Research in Biology (2013) 3(4): 947-953

Seino and Dongmo, 2013 16

Relative Chromosome Length (RCL)

16 14 12 10 8 6 4 2 0 L1

M9 S10 S11

14 12 10 8 6 4 2 0

Chromosome pair

A. turrita

S10 S11

Relative Chromosome Length (RCL)

C. compta

18 16 14 12 10 8 6 4 2 0 L1

S10 S11

14 12 10 8 6 4 2 0

Chromosome pair

S10 S11

Chromosome pair

O. spissus

C.s.producta 14

Relative Chromosome Length (RCL)

Chromosome pair

12 10 8 6 4 2

12 10

0 L1

S10 S11

8 6 4 2 0 L1

Chromosome pair

M. fasciata

P. luculenta Relative Chromosome Length (RCL)

16 14 12 10 8 6 4 2 0 L1

M9 S10 S11

Chromosome pair

S. pygmaeum Figure. 2: Idiograms of the seven species investigated Journal of Research in Biology (2013) 3(4): 947-953

951

Seino and Dongmo, 2013 r eport ed (Acrididae:

Neo-XY

Dichropl us

Melanoplinae).

This

type

v ittatus

Bidau CJ and Marti DA. 2000. Meiosis and the Neo-

XY system of Dichroplus vittatus (Melanoplinae:

sex

determination mechanism was absent in the seven species investigated in this study. The X-chromosome during this investigation was found to be medium in the four Acridinae. However, Chadha and Mehta (2011a), investigating Indian Acridinae observed that the X chromosome in A. turrita was the longest chromosome in the karyotype. There is

Acrididae): a comparison between sexes. Bugrov AG, Warchalowska-Sliwa E, MaryanskaNadachowska A. 1994. Karyotype evolution and chromosome C-banding patterns in some Podisma grasshoppers (Orthoptera: Acrididae) Caryologia, 47: 183-191.

therefore disagreement of this report with that of the

Bugrov AG. 1995. Interpopulation sex chromosome

present investigation. Chadha and Mehta (2011b)

polymorphism in the grasshopper Podisma sapprorensis

reported the X chromosome in Oedaleus abruptus

Shir. From Sakhalin and Kurile Island. Folia Biologica

(oedipodinae) to be the largest element in the karyotype.

(Krakow), 43: 51 - 53.

During the present study, the X-chromosome in P. luculenta and M. fasciata (Oedipodinae) were among the large chromosomes of the karyotypes. There is no doubt that the X chromosomes of different species of the Oedipodinae is one of the largest elements in the karyotype. Though this chromosome was acrocentric in

Bugrov AG

and Sergeev MG.

1997. A new

grasshopper species of the genus Podisma, Bertold (Orthoptera-Acrididae) from the Southern Island and its karyotypic features. Acta Zoologica Cracoviensia, 40: 47 -52.

the two Oedipodinae investigated here, Turkoglu and

Bugrov AG, Warchalowska E, Vysotskaya L. 1999.

Koca (2002) found the same chromosome in Oedipodia

Karyotypic features of Eyprepocnemidinae grasshoppers

schochi schochi and Acrotylus insbricus (Oedipodinae)

from Russia and Central Asia with reference to the B

from Turkey to be Metacentric in morphology.

chromosomes in Eyprepocnemis plorans (Charp.). Folia Biologica (Krakow), 47 (3 -4).

ACKNOWLEDGEMENTS The authors are thankful to Dr Watcho Pierre

Camacho JPM and Cabrero J. 1983. karyological

(Associate Professor in the Department of Animal

differences between two species of grasshopper genus

Biology, University of Dschang - Cameroon) for reading

Acrotylus (Acrididae: Oedipodinae). Caryologia, 36(2):

the manuscript and helpful suggestions. We are also

121-127.

grateful to Professor Mpoame Mbida (Head of the Laboratory of Applied Ecology (LABEA), University of Dschang for laboratory space.

Chadha P and Mehta A. 2011a. Chromosome study in few

species

(Acrididae:Tryxalinae):

karyotype analysis and distribution of constitutive heterochromatin.

REFERENCES

Acridids Journal

Entomology

and

Nematology, 3 (1): 14 â&#x20AC;&#x201C; 19.

Aswathanarayana NV and Ashwath SK. 2006. Structural polymorphism and C-banding pattern in a few

Chadha P and Mehta A. 2011b. Chromosome

Acridid grasshoppers. Cytology, 71(3):223 â&#x20AC;&#x201C; 228.

complement and C-banding patterns in 6 species of grasshoppers. International Journal of Genetics and Molecular Biology, 3(1): 25-30.

Journal of Research in Biology (2013) 3(4): 947-953

952

Seino and Dongmo, 2013 Channaveerappa

and

Ranganath

1997.

Karyology of few species of South Indian Acridids. II. Male germ line karyotypic instability in Gastrimargus. J. Biosci. 22 (3): 367 -374. Mayya S, Screepada KS, Hegde MJ. 2004. Nonbanded and C-banded karyotypes of ten species of shorthorned grasshoppers (Acrididae) from South India. Cytologia, 69(2): 167 - 174. Mestre J and Chiffaud J. 2009. Acridien du Cameroun et de Republique centrafricaine (Orthoptera Caelifera). Supplément au catalogue et atlas des acridiens d’Afrique de l’ouest. Edition numérique ISBN 978-2-9523632-1-1. Sandhu KS and Chadha P. 2012. Karyological studies of four species of grasshoppers from Gurdaspur district of Punjab, India. Nucleus, online first article, Vo 55(3):1 -4. Seino RA. 1989. Cytogenetic characterization of seven Acridomorphoid grasshoppers. MSc Dissertation, University of Lagos, Nigeria. Sharma T and Gautam DC. 2002. Karyotypic studies of eleven species of grasshoppers from north-western Himalayas. Nucleus, 45(1-2): 27-35. Turkoglu S and Koca S. 2002. Karyotype C- and G-band patterns and DNA content of Callimenus (=Bradyporus) macrogaster macrogaster. J. Insect Sci. 2 (24): 1-4. White MJD. 1973. Animal cytology and evolution. Third edition. Cambridge University Press. Yao S. 2006. The karyotypes and C-band of five Locusta species of Acridoidea in Guizhou. Journal of Guizhou Normal University (Natural Sciences), 2: 3 - 14.

Submit your articles online at www. jresearchbiology.com Advantages

Easy online submission Complete Peer review Affordable Charges Quick processing Extensive indexing You retain your copyright submit@jresearchbiology.com www.jresearchbiology.com/Submit.php.

953

Journal of Research in Biology (2013) 3(4): 947-953

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Impact of the residue of Deltamethrin and Endosulfan pesticides on biochemical toxicity and some neurotransmitter contents in different brain areas of male Albino mice Authors: Somaya M. Ismail1, Azza A. Said2, Samira M. El-Sayad2.

Institution: 1. Zoology department, Faculty of Science, Cairo university. 2. Zoology department, Faculty of Science, Fayoum university.

ABSTRACT: Evaluating the action of the residues of pesticides on non-target organisms has been of interest to many researchers. The present study aimed to evaluate the pesticides deltamethrin and endosulfan on biochemical toxicity and some neurotransmitter contents in different brain areas of male albino mice. The results showed that the daily oral administration of deltamethrin and endosulfan caused a significant decrease in neurotransmitter contents (NE, DA and GABA) in most of the tested brain areas (cerebellum, striatum, cerebral cortex, hypothalamus, brain stem and hippocampus). On the other hand a gradual significant reduction, ALT, AST and ALP enzyme activities, while the glucose level and acid phosphatase increase were observed in serum of mice treated with deltamethrin and endosulfan for two weeks. Also, this study has a significant inhibition in the activities of enzymes in liver tissues of treated mice including glutathione reductase. Meanwhile, the activity of lipid peroxide, glycolytic (PK, PFK and GPI) and gluconeogenic enzyme activities (F-1, 6-D-Pase) were significantly increased in liver tissues of treated mice in response to treatment. Additionally, total protein and glycogen content showed a significant reduction in liver tissues of mice treated with deltamethrin and endosulfan for two weeks. It was concluded that the pollution of the aquatic environment by deltamethrin and endosulfan pesticides, would adversely affect the metabolism of the mice.

Corresponding author: Somaya M. Ismail

Keywords: Deltamethrin, Endosulfan pesticide, Laboratory-bred strain Swiss albino male mice, neurotransmitter contents (NE, DA and GABA).

Email: mragaa11@yahoo.com

Article Citation: Somaya M. Ismail, Azza A. Said and Samira M. El-Sayad. Impact of the residue of Deltamethrin and Endosulfan pesticides on biochemical toxicity and some neurotransmitter contents in different brain areas of male Albino mice. Journal of Research in Biology (2013) 3(4): 954-966

Tel: +201118244212

Dates: Received: 12 Jan 2013

Accepted: 22 Feb 2013

Published: 24 May 2013

Web Address:

http://jresearchbiology.com/ documents/RA0324.pdf.

Journal of Research in Biology An International Scientific Research Journal

954-966 | JRB | 2013 | Vol 3 | No 4

www.jresearchbiology.com

Ismail et al., 2013 abundant in the environment and its use is increasing

INTRODUCTION Using pesticides is an important procedure for enhancing

agriculture

yield.

However,

the

(Pozo et al., 2006; Harner et al., 2006). It reaches aquatic

great

systems through direct application, as well as spray drift

consciousness, brought back upon their deleterious

and runoff from agricultural areas (Broomhall, 2002;

effects on human, animal and environmental health,

Jergentz et al., 2004 and Rand et al., 2010).

leading to the shortage of their use by imposing various rules (Ahmsd et al., 2010; Botella et al., 2004).

It is known that exposure to pesticides during development may interfere with the normal development

Among pesticides, Deltamethrin, which is a type

of neurotransmitter systems and cause their direct

II pyrethroids, has a wide acceptability, and is used in

damage (Richardson et al., 2006). The central nervous

agriculture

high

system (CNS) during development is particularly

activity against a broad spectrum of insect pests

susceptible to the toxic effects of xenobiotics (Tilson,

(Villarini et al., 1998). The oral route constitutes the

2000). The mechanism by which these effects occur is

main sources of general population exposure to this

not known but currently it is assumed that the

pesticide which is ingested within food and water

monoaminergic neurotransmitters play a role during

(Barlow et al., 2001).

development, defined as â&#x20AC;&#x153;morphogeneticâ&#x20AC;? (Buznikov

and

forestry

because

its

It has been reported that deltamethrin caused an

oxidative damage in liver and intestine of

Carassius auratus gibelio explained by an increase of

et al., 1996; Levitt et al., 1997; Nicotra and Schatten, 1990). Organophosphate pesticide represent one of the

LPO level and an enhancement of antioxidative defence

worldâ&#x20AC;&#x2122;s

most

parameters (Dinu et al., 2010). Oral absorption of

Consequently, many of its residues are frequently found

deltamethrin is rapid and is metabolized with microsomal

in the environment. The aim of this study was to

enzyme system in liver and with tissue esterase present

determine the effects of the pesticides, deltamethrin and

in intestinal wall and liver in addition to plasma

endosulfan

carboxylesterases (Usmani et al., 2006). According

neurotransmitter contents in different brain areas of male

to Simsek et al., (2008), Deltamethrin applied at different

albino mice.

commonly

used

biochemical

agrochemical.

toxicity

and

some

concentrations of 25, 50, 100, 200, 400, 800 and 1600 mg /L, for 1,24, 48, 72 and 96 h increased lipid

MATERIALS AND METHODS

peroxidation which is accompanied by a decrease of

Pesticides

reduced glutathione and catalase activity in digestive

Deltamethrin

gland and gill of fresh water mussel.

Deltamethrin is a synthetic pyrethroid pesticide

Endosulfan (6, 7, 8, 9, 10-hexachloro -1,5,5a,6,

[ ( S ) -a c ya n o-3 - ph en ox ybe n z yl ( 1 R, 3 R) - 3 -( 2, 2-

9a-hexahydro-6,9-methano -2, 4, 3 benzodioxathiepine-3

dibromovinyl)-2,2-dimethylcyclopropanecarboxylate]

-oxide) is a broad-spectrum organochlorine pesticide

with molecular formula (C22H19 Br2NO3). Solubility in

(insecticide and acaricide) first registered for use in the

water is <0.1 mg/L at 25oC. Relative molecular mass of

United States in 1954 to control agricultural insect and

the compound is 505.2 g/mol, and the melting point is

mite pests on a variety of fruits, vegetables, rice, grains,

100oC (Figure 1).

tea, coffee, cotton and also in animal farm and houses

Endosulfan

(US EPA). Results from a global monitoring network for

Endosulfan is an off-patent organochlorine

persistent organic pollutants revealed that endosulfan is

pesticide and acaricide that is being phased out globally

955

Journal of Research in Biology (2013) 3(4): 954-966

Ismail et al., 2013 [6, 7, 8, 9, 10, 10-Hexachloro-1, 5,5a, 6, 9, 9a-hexahydro

Animals

-6, 9-methano-2,4,3-benzodioxathiepine-3-oxide]. With

Swiss albino male mice of 10 weeks old with an

the molecular formula of (C9H6Cl6O3S). Solubility in

average weight of 28.5±2.5 g obtained from the National

water is 0.33 mg/L. Relative molecular mass is found to

Research Centre, Cairo, Egypt were used. They were

−1

be 406.93 g mol ,and the melting point is 70-100°C,

maintained in a well ventilated animal house. They were

343-373 K, 158-212 °F (Figure 1)

housed in large polypropylene cages with free access to food and water ad labium during the course of the experiment. Animals were housed in groups (5 animals/ group) and maintained under standard conditions of temperature (23°C to 25°C), a relative humidity of 65% to 86% and in a schedule of 12 hours of light and 12 hours of dark. Animal treatment The animals were divided into three groups (n=6) of equal number, The control group (1) was orally and daily administered with equivalent amount of the vehicle (distilled water) for two weeks, the second group was given drinking water with 1.28 mg/kg BW of

Fig. 1. Chemical structure of the pesticides Deltamethrin and Endosulfan

deltamethrin (Yousef et al., 2006) during two weeks of oral and daily administration and the third group was orally

and

daily

administered

with

endosulfan

(1.5 mg /kg BW). At the ends of the experimental period (2 weeks), the mice were sacrificed under diethyl ether anesthesia at fasting state. Effect of deltamethrin and endosulfan (pesticide) on some neurotransmitter contents in different brain areas of male albino mice During the experiment six mice of each group were decapitated each week till the end of the 2-week duration times. The mice were killed by sudden decapitation at the designed times. The brain was rapidly Fig. 2. Changes (%) of activities of glucose level (GL), some enzymes (alanine aminotransferase (ALT), aspartate aminotransferase (AST), Alkaline phosphatase (ALP), acid phosphatase (ACP) in serum of male mice,. Lipid peroxide (LP) glutathione (GSH) , pyruvate kinase (PK), phosphofructokinase (PFK), glucose phosphate isomerase (GPI), Fructose 1, 6-diphosphatase (F-1, 6-ase) enzymes, Total protein (TP), glycogen content in tissues of male mice liver exposed to LC25 of Deltamethrin and Endosulfan pesticides for 2 weeks.

and carefully excised and then dissected on dry ice glass plate according to the method of Glowinski and Lversen (1966) into the following regions; cerebellum, striatum cerebral

cortex,

hypothalamus,

brain

stem

and

hippocampus. Brain tissues were wiped dry with filter paper, weighed, wrapped in plastic films and then in aluminum foil and quickly frozen in dry ice. NE and DA were extracted and estimated in the brain tissues

Journal of Research in Biology (2013) 3(4): 954-966

956

Ismail et al., 2013 according to the method of Chang (1964) modified by

according to Bradford (1976) Determination of tissues

Ciarlone (1978). GABA were extracted and estimated in

glycogen was evaluated according to Nicholas et al.,

the brain tissues according to the method of Sutton and

(1956). Lipid peroxide ( LP) was measured according to

Simmonds (1973). The fluorescence was measured in

Buege and Aust (1978). Glutathione (GSH) was

Jenway 6200 fluorometer.

measured according to Moron et al., (1979) Pyruvate

Effect of deltamethrin and endosulfan (pesticide) on

kinase

biochemical toxicity of male albino mice

spectrophometrically by the method of Bucher and

Serum

samples

were

obtained

(PK)

relative

activity

was

measured

the

pfleiderer (1975). phosphofructokinase (PFK) was

centrifugation of blood of six rats of each group at

measured according to Zammit et al., (1978) Glucose

4000 rpm for 15 min at 4°C, and were then divided in to

phosphate isomerase (GPI) was measured according to

Eppendorf tubes. Isolated sera from each group were

King (1965). Fructose -1, 6-diphosphatase (F- 1, 6-ase)

stored at -20°C until they were used for the analyses. For

was measured according to Sand et al., (1980). All

preparation of tissue homogenates of mouse liver tissue

biochemical l parameters determined in this study were

of six mice of each group, one gram of liver tissues of

determined spectrophotometrically, using reagent kits

mouse from each group was homogenized in 5 ml

purchased from BioMerieux Company, France. Kits

distilled water at pH 7.5.A glass homogenizer was used

purchased from BioMerieux Company, France.

and the homogenate was centrifuged for 10 minutes at

Statistical analysis

3000 rpm, fresh supernatant was used.

The results obtained in the present work are

The levels of serum alanine aminotransferase

represented as means ± standard deviation (SD), and

(ALT), aspartate aminotransferase (AST) were measured

were analyzed using analysis of variance (ANOVA). The

according to Reitman and Frankel (1957). Alkaline

significance

phosphatase (ALP) was measured according to Belfield

calculated using the Duncan Multiple Range Test (Steel

and Goldberg (1971) and acid phosphatase (ACP) was

and Torrie, 1980).

difference

between

means

were

measured according to Wattiaux and De Duve (1956) and sera glucose concentrations (GL) were determined

RESULTS

according to the glucose oxides method of Trinder

Results in Table 1 showed that the daily oral

(1969). Total protein (TP) content was determined

administration of deltamethrin and endosulfan resulted in

Table (1): Effect of oral administration of Deltamethrin and Endosulfan on dopamine (DA) content in the different brain areas of male albino rat. Cerebellum mean ± S.E.

Striatum mean ± S.E.

Cerebral cortex mean ± S.E.

Hypothalamus mean ± S.E.

Brain stem mean ± S.E.

Hippocampus mean ± S.E.

122.7± 0.72

280.5±0.64

52.3±0.084

433 ± 4.2

310.2±0.45

222.1±0.6

82.3±1.2*

186.6±0.6*

36.5±0.21*

146.3±2.1**

254.5±1.4

160.2±0.62

32.79%

33.69%

30.21%

66.21%

17.69%

-27.87%

122.7± 0.72

280.5±0.64

52.3±0.084

433±4.2

310.2±0.45

222.1±0.6

48.2±2.42**

145.12±2.3**

21.4±0.73**

116.4±2.5***

192.63± 1.5**

112.6±1.6**

60.72%

48.26%

59.1%

73.12%

38.11%

49.30%

Pesticides

Deltamethrin

Endosulfan

- Statistical analyses were performed between control (C=6) and treated (T=6) animals by using paired t' test % : Percentage of change from control *p< 0.05,**p< 0.01 & ***p< 0.001 957

Journal of Research in Biology (2013) 3(4): 954-966

Ismail et al., 2013 a significant decrease in DA content in all brain area.

deltamethrin and endosulfan at the concentrations of

The maximal decrease (p<0.001) in DA content was

54.1% and 62.89%, respectively.

found in the hypothalamus of mice treated with

The results in Table 4 showed that a clear

deltamethrin and endosulfan at the concentrations of

reduction (P<0.001) in liver enzyme activities in serum

66.21% and 73.1%, respectively. Also, Table 2 showed

of mice treated with deltamethrin and endosulfan as

that the daily oral administration of deltamethrin and

compared to the control mice. On the other hand, the

endosulfan caused a significant (p<0.001) decrease in

glucose concentration and Acid phosphatase in serum of

GABA content in all the brain area, the maximal

treated mice showed a marked increase (P<0.001) in

decrease (p< 0.001) in GABA content was found in

comparison with the control group. Glycogen content in

brain stem of mice treated with deltamethrin and

tissues of treated mice showed a significant (p>0.001)

endosulfan at the concentration of 72.52% and 80.52%,

decrease in comparison with the control group. The

respectively.

reduction rates were 36.32% and 58.24% for mice

The results obtained from Table 3 showed that the maximal decrease (p<0.001) in NE content was

treated with deltamethrin and endosulfan, respectively (Table 7).

found in the hypothalamus of mice treated with Table (2):Effect of oral administration of Deltamethrin and Endosulfan on gama-butyric acid (GABA) content in the different brain areas of male albino rat. Cerebellum mean ± S.E.

Striatum mean ± S.E.

Cerebral cortex mean ± S.E.

Hypothalamus mean ± S.E.

Brain stem mean ± S.E.

Hippocampus mean ± S.E.

165.7±0.65

154.21±0.8

44.2±0.62

321.6±0.82

121.2±0.197

204.3±1.6

102.6±1.3**

92.6±0.428**

35.2±0.8

2925±0.43*

33.3±0.764**

98.8±0.577**

-61.97%

39.82%

18.1%

72.52%

51.64%

165.7±0.65

154.21±0.8

44.2±0.62

321.6±0.82

121.2±0.197

204.3±1.6

60.6±1.2***

72.4±0.87**

28.6±0.83***

252±1.6

23.6±0.82*

88.4±1.6***

63.43%

53.1%

35.29%

21.64%

80.52%

56.73%

Pesticide

Deltamethrin

Endosulfan

- Statistical analyses were performed between control (C=6) and treated (T=6) animals by using paired t' test. % : Percentage of change from control. *p< 0.05,**p< 0.01 & ***p< 0.001 Table (3): Effect of oral administration of Deltamethrin and Endosulfan on norepinephrine (NE) content in the different brain areas of male albino rat. Cerebellum mean ± S.E.

Striatum mean ± S.E.

Cerebral cortex mean ± S.E.

Hypothalamus mean ± S.E.

Brain stem mean ± S.E.

Hippocampus mean ± S.E.

102.6±1.4

434.2±1.6

64.6±1.54

462.2±2.11

342±0.53

233.1±1.4

77.5±0.56**

344.23±1.4**

35.2±0.54*

212.2±052**

243.1±0.45**

106.2±0.62***

24.64%

20.72%

48.57%

54.1%

28.95%

45.44%

102.6±1.4

434.2±1.6

64.6±1.54

462.2±2.11

342±0.53

233.1±1.4

48.2±0.62**

223.3±0.61**

22.3±1.4***

210.8±1.1***

168.5±1.4***

86.5±0.83***

53%

48.57%

65.48%

99.45%

50.73%

62.89%

Pesticide

Deltamethrin

Endosulfan

958

Ismail et al., 2013

Data represent mean values of five replicates. Within columns for dose, time and (dose x time), mean values followed by different letters are statistically significantly different based on LSD at P = 0.05.

37.27% 8.84 F±0.22 46.26% 1.94 ±0.01 49.52% 4.22 ±0.45 44.8% 10.04 ±0.311 64.9% 15.6 ±1.8 Endosulfan

11.8% 14.21 ±0.316 44.5 ±0.64 Deltamethrin

22.4 ±1.5

49.7%

% Change

18.21 ±0.432

44.87%

6.835 ±0.12

8.36 ±0.338

18.48%

2.85 ±0.05

3.61 ±0.03 % Change % Change AST

Control

959

7.21 ±0.22

6.44 ±0.23

11.96%

% Change % Change

ALP ALT

Liver function enzymes (umole/mg protein/min.) Glucose (GL) mg/g tissue

Table 4: Effect of Deltamethrin and Endosulfan on liver function enzymes in serum of male mice.

Acid phosphatase (ACP)

The present result in Table 5 indicated that a significant increase in lipid peroxide accompanied with a significant reduction in glutathione and total protein in liver enzyme activities of mice treated with deltamethrin and endosulfan as compared to the control mice. The present results in tables (6, 7) demonstrate a significant elevated level of glycolytic (PK, PFK and GPI) and gluconeogenic enzyme activities (F-1,6-DPase) in tissue of mice treated with deltamethrin and endosulfan as compared to the control. The elevation rates in the activities of PK, PFK, GPI and F-1-6, D-Pase enzymes were 97.36%, 76.1%, 74.84% and 69.1%, respectively for mice treated with endosulfan. DISCUSSION Many monoamine neurotransmitters, including DA , NE and GABA are important in the regulation of brain development prior to assuming their roles as transmitters in the mature brain (Whitaker-Azmitia, 1992; Di Pino, 2004; Ansorge, 2008), thus any circumstance that affects these neurotransmitters in the developing brain can alter the final structure and function of the brain. Developmental neurotoxicity involves alterations in behavior, neurophysiology. From the present results, it is clear that the daily oral administration of deltamethrin and endosulfan caused reducing side effect in some neurotransmitter tissue in the brain and a significant decrease in neurotransmitter contents (NE, DA and GABA) in most of the tested brain areas. Cerebellum which is responsible for the voluntary movement; pons and medulla oblongata which is responsible of essential reflexive acts; striatum which is a brain region responsible for motor activity; cerebral cortex is responsible for sensation including visual, auditory and olfactory as well as motor coordination and association, also is responsible for higher mental function such as thinking,

planning,

reasoning,

memory

and

consciousness and hippocampus, this is the key area Journal of Research in Biology (2013) 3(4): 954-966

Ismail et al., 2013 Table 5: Effect of Deltamethrin and Endosulfan on lipid peroxide, glutathione and total protein in male mice liver. Lipid peroxide(LP) (ug/g tissue)

Glutathione(GR) (ug/g tissue)

% change

Total protein(TP) (mg/ml)

% Change

% change

Control

0.65 ±0.01

30.22 ±1.22

52.44 ±2.11

Deltamethrin

0.95 ±0.57

-46.15%

20.20 ±1.12

33.16%

41.11 ±1.15

21.61%

Endosulfan

1.22±0.06

-87.69%

15.16 ±0.85

49.83%

31.22 ±1.65

40.46%

concerned with learning (Ansorge, 2008). Brain stem is

Locomotors activity as measured in the open field

responsible for integration of coordination of essential

appears to be associated with the dopaminergic system

reflexive acts such as swallowing, vomiting and

(Chiavegatto et al., 1998). Also, in the present study, we

respiration (Bloom, 1983).

similarly found a loss of the NE and gamma-butyric acid

Our findings support the idea that deltamethrin

(GABA) content in the cerebellum, striatum, cerebral

and endosulfan is neurotoxic in the developing brain.

cortex, hypothalamus, brain stem and hippocampus. The

The present result found that these pesticides induced a

loss of brain stem DA levels and the loss of hippocampus

decrease in DA levels in cerebellum, striatum, cerebral

GABA levels were higher in treated mice.

cortex, hypothalamus, brain stem and hippocampus of

These effects may represent a large number of

treated mice. The loss of hippocampus DA levels was

actions involved in the development of synaptic

higher in treated mice. DA is an important component of

dysfunction in these neurotransmitter systems that

the neuroendocrine mechanism that regulates the

ultimately

activation of male sexual behavior in mammalian species

Nevertheless further behavioral testing is needed to

(Castagna and Ball, 1997). Moreover, steroidogenesis in

confirm this suggestion. Moreover, the present findings

the brain may play a critical role in mammalian brain

might indicate that prenatal and postnatal exposure to

developmental of both sexes (Konkle and McCarthy,

pesticide altered the program for developmental of DA,

2011). Steroids play a role in the development of

NE, and GABA synaptic functions. Given that, the

catecholamines systems (Leret, 2009; Muneoka et al.,

dysfunction in serotonin and dopamine systems is

2010; Pappas et al., 2010).

involved such as appetite, affective, locomotion,

contribute

behavioral

anomalies.

It is known that DA is the major compound

learning, neurological and neuropsychiatric disorders

involved in the control of the motor system. Bernardi and

(Insel et al., 1990; Kaye, 2008), further testing of this

Palermo-Neto, (1983) showed that locomotion and

function is needed to confirm that alteration of these

rearing frequencies observed in an open field might be

neurotransmitter systems is the cause of some of these

used to detect drug-induced dopaminergic interference.

dysfunctions. In general, our results support the

Table 6: Effect of Deltamethrin and Endosulfan on some glycolytic enzymes in male mice liver. PK Control Deltamethrin Endosulfan

4.16 ±0.26 7.18 ±1.44 8.23 ±1.64

Glycolytic enzmes (umole/mg protein/min.) PFK % change % change 7.44 ±1.16 -72.6% 10. 1 ±1.22 -35.75% -97.36% 13.1 ±1.23 -76.1%

GPI % change 77.34 ±2.43 113.50 ±3.2 135.22 ±6.4

-46.81% -74.84%

Journal of Research in Biology (2013) 3(4): 954-966

960

Ismail et al., 2013 Table 7: Effect of Deltamethrin and Endosulfan on Glycogen and some Gluconeogenic enzymes in male mice liver. Fructose-1,6-diphos-phatase (umole/mg protein/min.)

Glycogen( mg/g tissue ) % change

% Change

Control

6.8 ±0.64

12.6±1.22

Deltamethrin

4.33±0.64

36.32%

15.4 ±1.11

22.22%

Endosulfan

2.84±1.02

58.24%

21.3 ±1.43

69.1%

suggestion that at least some of the effects of these

This was attributed to the irritation of liver cells by

disorders that are increasing in humans can be caused by

toxins or due to increase loss of intracellular enzyme by

exposure to neurotoxin environmental contaminants

diffusion through cell membrane. In the present study,

(Slikker W and Schwetz, 2003).

acid phosphatase show significant elevation in serum

In conclusion, the results observed in this study

of treated mice. Higher levels of acid phosphatase in

reinforce the idea of the use of neurochemical measures,

tissue was observed by El-Aasar et al., (1989) and

such as the DA, NE and GABA content and its

Abdel-Rahman et al., (1993), which was attributed to the

metabolites

irritation of liver cells by toxins or metabolic products of

neurotoxicity, including developmental neurotoxicity,

growing schistosomula of adult worms and eggs or due

induced by chemical agents. Because of serotonergic

to increase loss of intracellular enzyme by diffusion

dysfunction is involved in appetite and affective

through cell membrane which appear to act as a stimulus

disorders, and the catecholamine DA and NE have been

to the synthesis of more enzyme.

brain

regions

indicators

most often linked to the behavioral pathology of a

Regarding the sources of energy for mice,

number of neurological and psychiatric disorders, studies

deltamethrin and endosulfan significantly decreased the

of pesticide on DA, NE- and GABA. Related behaviors

glycogen content in liver tissues of treated mice, while

in animal models will be needed to clarify the outcomes

the glucose level increased in the serum of treated mice.

of long-term alterations in noradrenergic, serotonergic

This may be attributed to the activity of the pesticides

and dopaminergic systems identified here.

that impedes oxygen consumption of mice, thus inducing

Concerning, ALT, AST and ALP enzyme

anaerobic respiration. Under hypoxic conditions, animals

activities, gradual significant reduction was observed in

derive their energy from anaerobic breakdown of

serum of mice treated with deltamethrin and endosulfan

glucose, which is available to the cells by increased

for two week. The reduction observed in AST and ALT

glycogenolysis (Vincent et al., 1995; Sambasiva, 1999).

attributed to the hepatocellular damage resulting from

Nakano and Tomlinson (1967) have suggested that

chemical-toxicity,

levels

catecholamine levels rise under stressful environmental

showed an intimate relationship to cell necrosis and /or

conditions, enabling the increased utilization of glycogen

increased cell membrane permeability which led to the

for

discharge of enzyme to blood stream. The decrease in

requirements, the mouse has to increase the rate of

transaminase levels providing additional support for the

glycolysis thus bringing about a reduction of the

side effect of the deltamethrin and endosulfan on

glycogen content and increase glucose level in the blood

mitochondria of the hepatic cells as it is the subcellular

(Baskaran and Palanichamy, 1990;

localization of transaminases (El –Shazly et al., 2001).

Baskaran, 1990).

961

where

the

transaminases

energy

production.

restore

its

energy

Vasanthi and

Journal of Research in Biology (2013) 3(4): 954-966

Ismail et al., 2013 The data obtained in the present study showed

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966

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Prevalence and the effect of plant extracts on community associated methicillin resistant Staphylococcus aureus in Owerri, Imo State, Nigeria Authors: Amadi ES1, Oguoma OI1, Ibekwe VI1, Abanobi SE2, Chikwendu CI1 and Egbadon OE1.

ABSTRACT: The prevalence of Methicillin resistant Staphylococcus aureus (MRSA) among apparently healthy inhabitants of Eziobodo Community and Students of Federal University of Technology Owerri (FUTO), Imo State, Nigeria was studied. The work further ascertained the antibacterial activities of medicinal plants including Azadirachta indica, Pterocarpus mildbraedii, Garcinia kola, Phyllanthus amarus and Vernonia amygdalina against the MRSA isolates. A total of two hundred nasal swab specimens were randomly collected from the participants. The Kirby-Bauer technique was used to determine the susceptibility pattern of the isolates to Vancomycin (5µg), Institution: Ciprofloxacin (5µg), Ceftriaxone (30µg), Oxacillin (5µg), Methicillin (10µg) and 1. Department of Erythromycin (15µg). The antibacterial properties of the ethanolic plant extracts were Microbiology, School of Science, Federal University determined using the agar well diffusion technique. A total of 181 (90.5%) and 141 of Technology, P.M.B. 1526, (70.5%) of the nasal swab samples, yielded Staphylococcus species and Staphylococcus aureus respectively. The antibiotic sensitivity screening revealed that Owerri Imo State, Nigeria. 38 (27%) of the S. aureus isolates were methicillin resistant. The MRSA isolates also 2. Department of exhibited the highest resistance to vancomycin and the least to ceftriaxone. Biochemistry, School of Furthermore, the result showed that crude ethanolic extracts of all tested plant Science, Federal University extracts except Pterocarpus mildbraedii exhibited antibacterial activities against the of Technology, P.M.B. 1526, MRSA isolates. Phytochemical components such as Alkaloids, Tannins, Glycosides, Owerri Imo State, Nigeria. Saponins, Flavonoids, Terpenoids, Phlobatannins, Steroids and Anthraquinones were detected in the plant materials in varying proportions. This study unveils a relatively high occurrence of MRSA among the study population which could be a risk factor for infection with MRSA. These plant extracts could also serve as potential sources of therapy for the treatment of MRSA infections. Corresponding author: Chikwendu CI.

Keywords: Antibiotic resistance, Staphylococcus aureus, Methicillin, Plant extracts, Isolates.

Email:

Article Citation: Amadi ES, Oguoma OI, Ibekwe VI, Abanobi SE, Chikwendu CI and Egbadon OE. Prevalence and the effect of plant extracts on community associated methicillin resistant Staphylococcus aureus in Owerri, Imo State, Nigeria. Journal of Research in Biology (2013) 3(4): 967-976 Dates:

Web Address:

Received: 20 Mar 2013

Accepted: 09 May 2013

Published: 05 June 2013

http://jresearchbiology.com/ documents/RA0342.pdf. This article is governed by the Creative Commons Attribution License (http://creativecommons.org/ licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.

Journal of Research in Biology An International Scientific Research Journal

967-976 | JRB | 2013 | Vol 3 | No 4

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Amadi et al., 2013 and Daum (2010), all infections occurring among

INTRODUCTION Staphylococcus aureus is a coagulase positive,

outpatient or among inpatients with an MRSA obtained

gram positive cocci, which apart from being a normal

earlier than 48 hours after hospitalization could be

flora of the anterior nares, skin and large intestine, is also

regarded as CA-MRSA. In addition, livestock associated

capable of causing a wide range of diseases varying from

MRSA (LA-MRSA) have been reported to pose a

minor skin infections to life threatening septicemia,

challenge particularly in countries with low level of

pneumonia, endocarditis, deep-seated abscess among

MRSA (Stefani et al., 2012). Morris et al.,

others (Willey et al., 2008; Lowy 2003; Kuehnert et al.,

reported the potential for pet animals to harbour MRSA

2006; Tenover and Gaynes, 2000; Holmes et al., 2005;

when residing with human MRSA patients.

(2012)

Nester et al., 2007). Penicillin and later methicillin were

The fact that MRSAs are becoming more

very efficacious in the management of Staphylococcus

prevalent worldwide and also resistant to a wide range of

infections in the early 1960s. However, over the years,

antibiotic groups, underlines the need for alternate

most strains have acquired resistance to these drugs due

strategies to stem the immense public health challenge

to acquisition of gene encoding the enzyme penicillinase.

posed by these organisms. Natural products from local

In recent times, strains of S. aureus have emerged that

medicinal plants are increasingly being used in the

not only produce penicillinase, but also have Penicillin

treatment of many hard to treat diseases and the search

binding proteins (PBPs) with low affinity for all β-lactam

for more potential compounds from plants has continued

drugs. These strains referred to as methicillin resistant

(Lai et al., 2010; Newman and Cragg, 2007). According

Staphylococcus

to WHO, 65-80% of the world population rely on

drugs

traditional medication for their ailments (Gurinder and

(Nester et al., 2007; Willey et al., 2008). Nearly all

Daljit, 2009). A number of works has highlighted the

MRSA have additional genetic material known as mec A

efficacy of local indigenous plants against a wide range

gene not found in methicillin sensitive strains, which

of pathogens (Ugbogu et al., 2010; Lai et al., 2010;

encodes PBP 2a, a cell wall transpeptidase, having

Aliyu et al., 2008; Aliyu et al., 2011; Ajibade et al.,

reduced affinity for β -lactam antibiotics. The mec A

2010). The present study was aimed at ascertaining the

gene is found as a part of a mobile genetic element found

occurrence

in MRSA strains known as Staphylococcal cassette

Eziobodo community inhabitants and FUTO students as

chromosome mec (SCC mec) (Jeshina and Surekha,

well as their susceptibility to different antibiotic groups.

2009; Pinho et al., 2001).

It also determined the antibacterial effects of some local

m eth i cil l in

aureus an d

(MRSA)

oth er

are

resistant

-l a cta m

In addition to β lactam drugs, MRSA isolates

of MRSA

among apparently healthy

plant extracts on the MRSA isolates.

have become resistant to a number of antimicrobial agents such as, fluoroquinolones, aminoglycosides and

MATERIALS AND METHODS

macrolides (Shittu et al., 2009). MRSA could be

Collection of nasal specimens

categorized as either hospital acquired (HA-MRSA) or

Two hundred (200) nasal specimens were

community acquired (CA-MRSA), depending on the

collected, 100 each from the anterior nares of apparently

source of acquisition. While the former occur in

healthy individuals of Eziobodo community (one of the

individuals who are/have recently been in a hospital or

communities hosting FUTO) and students of Federal

other healthcare facility, the latter are acquired by

University of Technology (FUTO), all in Owerri West

persons not recently hospitalized. According to David

LGA, Imo State, Nigeria. They were aseptically

968

Journal of Research in Biology (2013) 3(4): 967-976

Amadi et al., 2013 collected using sterile swab sticks between August and November 2010. Cultivation and isolation of Staphylococcus aureus The respective nasal specimens were cultivated within one hour of collection in Mannitol salt agar and nutrient agar using standard techniques to obtain discreet colonies. The plates were incubated at 37oC for 24 hours. The axenic cultures of the isolates were subsequently identified using colony morphology, microscopy and biochemical tests including catalase and coagulase tests (Cheesbrough, 2002). Antibiotic susceptibility test The antibiotic susceptibility screening of the S. aureus isolates was conducted using the Kirby-Bauer

Figure 1: Antimicrobial resistance rates (%) of MRSA isolates to different antibiotics Preparation of plant extracts

disc diffusion method (Cheesbrough, 2002). Standard

The leaves, barks and seeds of the plants were

inoculum, equivalent of 0.5 McFarland standards of the

washed and dried at room temperature and later

isolates was evenly spread on Mueller Hinton agar

pulverized. 20gm of each plant powder was separately

plates. Antibiotic discs including Vancomycin (5µg),

mixed with 250ml of ethanol and the extraction was done

Ciprofloxacin (5µg), Ceftriaxone (30µg), Oxacillin

using the soxhlet extraction procedure.

(5µg), Methicillin (10µg) and Erythromycin (15µg)

Phytochemical screening

(Oxoid) were aseptically placed on the plates. The plates

The phytochemical screening of each plant

were then incubated at 37oC for 24 hours and the

extract was carried out to determine the presence or

inhibition zones recorded in millimeters using a meter

absence of Alkaloids, Tannin, Saponins, Glycosides,

rule.

Anthraquinone, Steroids, Flavonoids, Terpenoids, and Subsequently, all the isolates identified as

phlobatannins (Harbone, 1973; Sofowora, 1993).

Methicillin resistant Staphylococcus aureus (MRSA)

Susceptibility screening of MRSA isolate to plant

were subjected to antibiotic screening test using the same

extracts

disc diffusion technique as above. The following

The antibacterial effects of each plant extract on

antibiotics were used; Vancomycin (5µg), Ciprofloxacin

MRSA were determined using the agar well diffusion

(5µg), Ceftriaxone (30µg) and Erythromycin (15µg)

technique (Perez et al., 1990). Standard inoculum,

(Oxoid).

equivalent of 0.5 McFarland standards of the isolates was

Collection of plant materials

evenly spread on Mueller Hinton agar plates. Sterile cork

The leaf and bark of Pterocarpus mildbraedii,

borer was used to make wells on the agar. The

Azadirachta indica, leaves of Vernonia amygdalina, and

reconstituted extracts (25mg/ml, 50mg/ml, 100mg/ml

whole plant of Phyllanthus amarus were obtained from

and 200mg/ml) were respectively introduced into wells

the premises of FUTO. The seeds of Garcinia kola

and labeled accordingly. Following the incubation of the

however, were purchased from Ekeukwu Owerri market,

plates at 37oC for 24 hours, the inhibition zone diameters

Imo State. The plant materials were subsequently

were recorded using meter rule.

authenticated by a taxonomist. Journal of Research in Biology (2013) 3(4): 967-976

969

Amadi et al., 2013 Table 1: Prevalence (%) of Staphylococcus aureus and MRSA isolates from Eziobodo and FUTO inhabitants Target population

No of samples

Staphylococcus sp.

Staphylococcus aureus

MRSA

Eziobodo

100

95(95)

66(66)

20(30.3)

FUTO students

100

86(86)

75(75)

18(24)

Total

200

181(90.5)

141(70.5)

38(26.9)

The Methicillin resistant Staphylococcus aureus

RESULTS AND DISCUSSION Out of the total of 200 specimens collected, 181

(MRSA) prevalence rate of 27% among apparently

isolates were identified as Staphylococcus species, while

healthy individuals as reported in the present work is

141 isolates were identified as S. aureus, representing a

considerably low compared to a report by Ugbogu et al.,

prevalence rate of 90.5% and 70.5% respectively

(2010) who isolated 83.5% of MRSA from healthy

(Table 1). Also, the antibiotic resistance screening of the

individuals in Abia State, South East Nigeria and

isolates showed that 38 (27%) of the S. aureus isolates

Onanuga et al., (2005) that recorded 69% isolation from

were MRSA. The MRSA isolates exhibited their highest

healthy women in Zaria, Nigeria. Similarly, our current

sensitivity to Ceftriaxone and the least to Vancomycin

finding is also lower than the report of Olowe et al.,

antibiotic (Table 2 and Figure 1).

(2007) and Onanuga and Temedie (2011), in which

The Phytochemical screening of the plant extracts revealed the presence

47.8% and 47.6% MRSA were isolated in South West

of Phytochemical

and Niger Delta regions of Nigeria respectively. The

components such as alkaloids, saponins, flavonoids and

prevalence rate of 47.15% and 43% has also been

others in varying quantities (Table 3).

reported from Ibadan and Jos, Nigeria (Ghebremedhin

The antibacterial screening of the ethanolic

et al., 2009; Ekeh, 2003). However, the current result is

extracts of the medicinal plants used in this study

consistent with the report of Nwankwo et al., (2010) in

indicated that all except Pterocarpus mildbraedii,

which 28.6% was recorded. The difference in the

exhibited inhibitory activity against MRSA isolate.

prevalence of MRSA obtained in the present study and

(Table 4).

those of previous works could be attributable to strain

The result of this study showed that 90.5% and 70.5% of the isolates from the anterior nares of Eziobodo

variation in different geographical regions and locations (Ikeagwu et al., 2008).

inhabitants and FUTO students were respectively

It is important to note that the recovery of MRSA

Staphylococcus species and Staphylococcus aureus. This

from apparently healthy community inhabitants in the

is consistent with the findings of Ugbogu et al., (2010)

present study is very significant particularly at this time

and Chigbu and Ezeronye (2003), but higher than the

when infiltrations of Community acquired MRSA

33.3% prevalence reported from Amassoma community

(CA-MRSA) to healthcare facilities has been reported in

in Niger Delta, Nigeria (Onanuga and Temedie,

some parts of the world (Stefani et al., 2012). According

2011).This high occurrence in our present work is not

to Creech et al., (2005), Farley et al., (2008), and

unexpected, since S. aureus is a normal microflora of the

Hidron et al., (2005) enormous reservoirs of MRSA now

human body, particularly the upper respiratory tract

exist outside health care settings and this implies that the

(Willey et al., 2008; Cheesbrough 2002).

current methods of MRSA control in health facilities are not likely to succeed. In this regard, preventive measures

970

Journal of Research in Biology (2013) 3(4): 967-976

Amadi et al., 2013 Table 2: Frequency (%) of antibiotic resistance S. aureus isolates from nasal samples of Eziobodo and FUTO inhabitants No (%) of resistant isolates Eziobodo FUTO No of isolates

No of resistant isolates

No of isolates

No of resistant isolates

Total no of isolates

Oxacillin

21(31.8)

20(26.7)

141

Total no of resistant isolates 41(29.1)

Methicillin

18(27.3)

20(26.7)

141

38(27)

Ciprofloxacin

16(24.2)

10(13.3)

141

36(25.5)

Vancomycin

15(22.7)

7(9.3)

141

22(15.6)

Erythromycin

10(15.1)

9(12)

141

19(13.5)

Ceftriaxone

3(4.5)

0(0)

141

3(2.1)

Antibiotics

Table 3: Phytochemical components of plant extracts Phytochemical components Plant extracts

AIL AIB PML PMB PA VA GA

Alkaloids

Tannins

Glycosides

Saponinss

Flavonoids

Phlobatannins

Steroids

Anthraquinones

Terpenoids

+ + + -

+ + + + +

+ + + -

+ + + + +

+ + + + -

+ + + +

+ + + -

+ + + + + +

+ + +

Key: AIL – Azadirachta indica Leaf, AIB – Azadirachta indica Bark, PA – Phyllanthus amarus, PML – Pterocarpus mildbraedii Leaf, PMB – Pterocarpus mildbraedii Bark, VA – Vernonia amygdalina, GK – Garcinia kola.

to stop the possible transmission in the communities is a

the over 70% resistance recorded by Ojulong et al.,

viable approach ( Charlebois et al., 2004; Cooper et al.,

(2009) in Kampala, Uganda. Azeez-Akande et al.,

2004; David et al., 2008; Liu et al., 2008).

(2008), however reported a susceptibility rate of 93.9%

The antibiotic susceptibility test revealed that all

of MRSA isolates to ciprofloxacin.

the S. aureus isolates exhibited the least resistance to

Furthermore, vancomycin has been described as

ceftriaxone antibiotic in the present study. This finding is

a reliable alternative for the treatment of MRSA

consistent with the report of Masood and Aslam (2010)

infections. Elhamzaoui et al., (2009) and Nwankwo and

in which 96.1% susceptibility of S. aureus isolates to

Nasiru (2011) reported 100% sensitivity of S. aureus

ceftriaxone was highlighted. Ceftriaxone was apparently

isolates from a University hospital in Rabat Morocco and

recommended by these workers as a drug of choice for

a tertiary health institution in Kano, Nigeria, to

infections caused by S. aureus, Escherichia coli,

Vancomycin respectively. Nevertheless, this antibiotic,

Pseudomonas aeruginosa, Klebsiella pnuemoniae and

vancomycin, which was initially a drug of choice in the

Salmonella typhi. On the other hand, the S. aureus

treatment of MRSA infections, is witnessing resistance

isolates were more resistant to Ciprofloxacin and

in recent times (Von-Eiff et al., 2001). In the present

Erythromycin (Tables 2 and 3). The resistant rates are in

work, over 50% of the MRSA isolates were resistant to

line with the reports of Shanhraz et al., (2012), and

Vancomycin. This is worrisome because Vancomycin

Onanuga and Temedie (2011), but quite low compared to

has been described by various workers as very effective

Journal of Research in Biology (2013) 3(4): 967-976

971

Amadi et al., 2013 concomitant effect of the active compounds contained by

Table 4: Inhibitory activities of plant extracts against MRSA isolates

these plants on MRSA. However, none of the extracts

Mean zone of inhibition (mm)/ Concentration of plant extracts (mg/ml) Plant extract AIL AIB PML PMB PA VA GK

100

200

7 -

8 9 -

9 12 8 10 11

were active against the MRSA isolates at the lowest concentration of 25mg/ml (Table 4). The inhibitory effect of Garcinia kola extract on MRSA as observed in the present study is in agreement with the work of Ugbogu et al., (2010) and Adeleke et al., (2006), in which Garcinia kola extracts exhibited antibacterial activities against MRSA isolates in Nigeria. Also, Taiwo et al., (1999) reported that Garcinia kola exhibited strong activity against MRSA.

Key: AIL – Azadirachta indica Leaf, AIB – Azadirachta indica Bark, PA – Phyllanthus amarus, PML – Pterocarpus Similarly, the inhibitory properties of Azadirachta indica mildbraedii Leaf, PMB – Pterocarpus mildbraedii Bark, and Vernonia amygdalina against MRSA as recorded in VA – Vernonia amygdalina, GK – Garcinia kola.

the present work is consistent with the reports of against MRSA and in fact a drug of choice in the

Skariyachan et al., (2011), Aliyu et al., (2011)

treatment of multidrug resistant S. aureus infections

and

(Ojulong et al., 2009; Elhamzaoui et al., 2009). The

Phyllanthus amarus extract had antibacterial activity

vancomycin resistance rate as recorded in the current

against MRSA is in line with the findings of

study is however contrary to the report of Onanuga and

Aliyu et al., (2008). Ajibade et al., (2010) also

Temedie

and

highlighted the antimicrobial activity of Phyllanthus

Shanhraz et al., (2012) in which over 70% susceptibility

species against MRSA. Undoubtedly, the findings of this

was recorded. The present finding thus suggests that

study support the local use of these plant materials in the

vancomycin may be inefficient in the treatment of

treatment of most hard to treat infections.

(2011)

Niger

Delta

Nigeria

Aliyu

infections caused by MRSA in the near future among our target

population.

The

present

study

al.,

(2008).

Furthermore,

that

In conclusion, the recovery of CA-MRSA from

therefore

the external nare of apparently healthy individuals in this

recommends ceftriaxone as a drug of choice for the

study underscores the significance of the nasal region as

treatment of MRSA infections in our study area.

a reservoir of S. aureus, and by implication MRSA. In

The increasing resistance of MRSA to β-lactam

fact, MRSA colonization of the nares is believed to be a

and other broad spectrum antibiotics has stimulated

risk factor for a clinically apparent infection with MRSA

recent investigations on plant parts for naturally

(Croft et al., 2009; Huang et al., 2006; Lu et al., 2007;

occurring active compounds as alternatives to treatment

Muder et al., 1991). It is therefore very imperative that

of MRSA

strategies should be designed to halt the further spread of

caused infections. The phytochemical

screening of the plant extracts used in this study revealed

MRSA

the presence of alkaloids, Tannins, saponins, flavonoids,

immunodeficient

terpenoids, anthraquinones, glycosides and steroids

Stefani et al., (2012), CA-MRSA clones spreading in the

(Table 4). The antibacterial screening of the plant

community could also infiltrate healthcare facility in

extracts showed that all the plant materials used except

many parts of the world. This certainly would exacerbate

Pterocarpus mildbraedii exhibited inhibitory activity

the challenges already posed by MRSA. Interestingly

against MRSA. This effect could be attributed to the

however, the therapeutic activities of the plant materials

972

communities

and most

individuals.

especially to

According

Journal of Research in Biology (2013) 3(4): 967-976

Amadi et al., 2013 used in this study could hold a great promise as a

Chigbu CO and Ezeronye OU. 2003. Antibiotic

potential precursor in the development of therapies for

resistant Staphylococcus aureus in Abia State of Nigeria.

the management of MRSA infections, if properly

Afr. J. Biotechnol., 2(10): 374-378.

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Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Odonata diversity (Insecta: Arthropoda) in rice and vegetable fields in a north-eastern district of Tamil Nadu, India Authors: Veeramuthu Anbalagan, Michael Gabriel Paulraj and Savarimuthu Ignacimuthu*

Institution: Entomology Research Institute, Loyola College, Chennai-34.

ABSTRACT: Odonata diversity in vegetable fields (brinjal and okra) and rice fields was studied from January 2005 to December 2008 in Tiruvallur district of Tamil Nadu. Totally 23 species of Anisoptera (dragonflies) and 12 species of Zygoptera (damselflies) were recorded and all these species were grouped into eight families. In vegetable fields 31 species of dragonflies and damselflies were recorded under 22 genera. In rice fields the species richness (21 species) and total genera (16) were less than vegetable fields during the entire study period. Libellulidae was the large family in both vegetable and rice fields which comprised maximum number of species. Pantala flavescens (Fabricius), a migratory species, was the most dominant in numbers throughout the year. Diversity indices clearly showed that odonata diversity was higher in vegetable fields than in rice fields.

Corresponding author: Keywords: Savarimuthu Ignacimuthu Dragonflies, Damselflies, Libellulidae, Pantala flavescens

Email Id:

Web Address:

http://jresearchbiology.com/ documents/RA0346.pdf.

Article Citation: Veeramuthu Anbalagan, Michael Gabriel Paulraj and Savarimuthu Ignacimuthu. Odonata diversity (Insecta: Arthropoda) in rice and vegetable fields in a north-eastern district of Tamil Nadu, India. Journal of Research in Biology (2013) 3(4): 977-983 Dates: Received: 06 Apr 2013

Accepted: 23 May 2013

Published: 05 June 2013

Journal of Research in Biology An International Scientific Research Journal

977-983 | JRB | 2013 | Vol 3 | No 4

www.jresearchbiology.com

Anbalagan et al., 2013 INTRODUCTION

Sampling of Odonates:

Dragonflies and damselflies in the order Odonata

In each village, dragonflies and damselflies were

are important group of insects in agroecosystems, forest

sampled in three different locations by quadrate method.

ecosystems and aquatic ecosystems. They are potential

Quadrates of 25 m x 10 m size were laid down with

biocontrol agents of agricultural, horticultural and forest

threads inside rice, brinjal and okra fields separately.

pests. Many studies have shown that the larval stages of

Totally three quadrates were put in each rice and

Odonata are important biological control agents of

vegetable fields. Perched dragonflies and damselflies

mosquito larvae (Mandal et al., 2008; Spencer et al.,

found inside the quadrates were collected by sweeping

1999).

According to Corbet (1999), dragonflies and

net (25 cm in diameter) during day times (between 10.00

damselflies are excellent ecological indicators. Around

AM to 15.00 PM). Flying Odonates inside quadrate area

6,000 species and subspecies of Odonata have been

were also caught with sweeping net. Sampling was done

described under 630 genera in 28 families throughout the

twice in a month from January 2005 to December 2008.

world (Tsuda, 1991). In India, 499 species, 139 genera

Specimens from replications were pooled together.

and 17 families of dragonflies and damselflies have been

Identification:

documented (Prasad and Varshney, 1995; Sharma,

The

specimens

were

identified

using

2010). Odonata diversity has been extensively studied in

identification keys provided by Fraser (1933, 1934 and

different forest areas. Emiliyamma (2005) has recorded

1936) and Subramanian (2009). After identification and

31 species of dragonflies and damselflies from southern

counting the total number of specimens, few specimens

Western Ghats in the Kottayam district of Kerala. Very

from each taxa were retained and others were left behind

few investigators have studied the Odonata diversity in

alive in the field. Specimens which were not identified in

agricultural

1999;

the field were brought to the laboratory for identification.

A knowledge on Odonata

The identified specimens were deposited at the

fields

(Gunathilagaraj

Kandibane et al., 2005).

al.,

diversity in different agro ecosystems is very essential to

Entomology

understand the influence of crop type on species

Chennai.

richness, abundance and evenness of dragonflies and

Meteorological Data:

damselflies. Hence the present work was undertaken to

Data

Research

Institute,

atmospheric

Loyola

College,

temperature, relative

assess the Odonata diversity in two different agricultural

humidity, mean total rainfall and total number of rainy

fields, i.e. rice and vegetable fields in Tiruvallur district

days from 2005 to 2008 were obtained from Regional

of Tamil Nadu.

Meteorological Centre, Chennai. Diversity indices:

MATERIAL AND METHODS Study site:

Total number of dragonflies and damselflies collected during the study period was recorded. Total

Dragonflies and damselflies were sampled from

abundance, Simpson‟s index of diversity (1-D), Shannon

vegetable fields, viz. brinjal and okra in Kolappancheri

-Wiener Diversity Index (H), Shannon entropy, species

village and rice fields in Vayalanallur village of

richness and species evenness were calculated by using

Tiruvallur district. The geocordination of Tiruvallur

the software „Past.exe‟ (ver. 2.14). Jaccard‟s similarity

district is 12° 15 and 13° 5`N Latitude and 99° 15` and

index was calculated to find out the similarity in Odonata

80° 20` E Longitude.

diversity between vegetable and rice fields.

978

Journal of Research in Biology (2013) 3(4): 977-983

Anbalagan et al., 2013 The formulae for the diversity indices are as follows: Simpson‟s index (D) = Σ ni (ni-1) / N (N-1) i=1 Where ni = number of individual for each species N = total number of individuals Shannon index of general diversity ( H )

ni N

log

ni N

Where ni = number of individual for each species N = total number of individuals

Figure 1. Total number of genera and species collected under different families of Odonata collectively from vegetable and rice fields

Evenness (e) e=

H log S

Coenagrionidae, Platycnemididae

were

recorded

Lestidae under

and

Zygoptera

(damselflies) (Table 1). Totally 31 species of dragonflies

Where H = Shannon index

and damselflies were recorded under 22 genera, of which

S = number of species The similarity in odonata diversity between vegetable fields and rice fields was assessed by using the formula of Jaccard‟s similarity index as follows: Jaccard‟s Index = A/ (A+B+C) Where A= total number of species present in both communities B= the number of species present in community 1 but not 2 C= the number of species present in community 2 but not 1

15 genera and 22 species were dragonflies and 7 genera and 9 species were damselflies. Libellulidae was found to be the largest family, which has the highest number of species (18 species) throughout the study period. Species richness was 31 throughout the study. Total abundance was maximum (4167) in 2008. Maximum evenness of 0.899 was recorded in vegetable fields in 2007 and this was correlated with the maximum Shannon-Wiener diversity index of 3.328 during the same study year (Table 2). The similarity index (Jaccard‟s similarity

RESULTS Totally 35 species of dragonflies and damselflies were recorded collectively from vegetable and rice fields in Tiruvallur district from January 2005 to December 2008 (Figure1). The species composition, richness, evenness and other diversity indices showed variations between vegetable and rice fields. Species composition and diversity in vegetable fields Three families viz., Aeshnidae, Gomphidae and Libellulidae

Euphaeidae,

were

recorded

under

Anisoptera

(dragonflies) and five families viz., Calopterygidae, Journal of Research in Biology (2013) 3(4): 977-983

index) was calculated as 0.660 for each study year (Table 2). Species composition and diversity in rice fields Five

different

families namely Aeshnidae,

Gomphidae, Libellulidae, Coenagrionidae and Lestidae were recorded in rice field. All the species collected from rice fields were grouped under 16 genera (12 Anisoptera and 4 Zygoptera). Total number of species recorded in rice field was 21 (15 Anisoptera and 6 Zygoptera). Maximum total abundance (1703) was recorded in 2008. Maximum Shannon-Wiener diversity index (2.871) and 979

Anbalagan et al., 2013 Table 1. Taxonomic composition and total number of individuals collected under different species of Odonata from North-Eastern Tamilnadu during 2005-2008 Sl.No.

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35

Species Anisoptera Family: Aeshnidae Anax guttatus (Burmeister) Anax immaculifrons (Rambur) Family: Gomphidae Heliogomphus selysi (Fraser) Ictinogomphus distinctus (Rambur) Ictinogomphus rapax (Rambur) Family: Libellulidae Brachythemis chalybea (Brauer) Brachythemis contaminata (Fabricius) Bradinopyga geminata (Rambur) Crocothemis servilia (Drury) Diplocodes trivialis (Rambur) Neurothemis tullia (Drury) Orthetrum glaucum (Brauer) Orthetrum sabina (Drury) Orthetrum testaceum (Burmeister) Pantala falvescens (Fabricius) Rhyothemis variegata (Linn.) Sympetrum vulgatum flavum (Bartenef) Tholymis tillarga (Fabricius) Tramea basilaris (Palisot de Beauvois) Tramea limbata (Desjardins) Trithemis aurora (Burmeister) Trithemis festiva (Rambur) Trithemis pallidinervis (Kirby) Zygoptera Family:Calopterygidae Caliphaea sp Family: Coenagrionidae Agriocnemis femina femina (Brauer) Agriocnemis pygmaea (Rambur) Ceriagrion coromandelianum(Fabricius) Ischnura aurora (Brauer) Ischnura delicata (Hagen) Ischnura inarmata (Calvert) Ischnura senegalensis (Rambur) Family: Euphaeidae Euphaea sp Family: Lestidae Lestes viridulus (Rambur) Family: Platycnemididae Copera marginipes (Rambur) Platycnemis sp Total

Number of individuals collected Vegetable fields Rice fields 2005 2006 2007 2008 2005 2006 2007

2008

0 25

0 52

0 67

0 72

9 12

11 6

7 14

6 11

179 128 112

158 94 75

124 108 82

186 134 92

38 0 29

88 0 42

76 0 37

54 0 32

128 106 27 220 175 98 116 125 114 480 219 90 30 170 150 112 107 72

142 85 35 145 205 112 105 145 108 306 184 109 18 165 120 78 118 110

108 122 42 189 218 147 98 102 122 318 225 128 45 138 111 65 128 95

129 148 33 238 232 121 165 148 148 372 236 114 55 145 165 92 108 108

0 78 0 36 125 58 78 51 0 185 89 0 0 31 0 26 0 52

0 55 0 42 163 82 67 25 0 211 58 0 0 27 0 34 0 45

0 86 0 46 158 117 63 48 0 203 62 0 0 41 0 31 0 69

0 73 0 58 182 93 85 60 0 197 71 0 0 29 0 40 0 42

0 0 190 70 0 71 92

0 0 78 78 0 65 84

0 0 158 65 0 108 149

0 0 212 128 0 108 132

110 92 140 43 121 0 0

74 68 61 59 82 0 0

101 81 125 78 88 0 0

122 105 156 88 106 0 0

118

120

70 54 3656

78 65 3277

92 88 3670

55 65 4167

0 0 1464

0 0 1368

0 0 1622

0 0 1703

evenness (0.8409) in rice fields were recorded during

similarity index) was calculated as 0.660 for each study

2007. Odonata diversity in rice fields was lower than

year.

vegetable 980

fields.

The

similarity index (Jaccardâ&#x20AC;&#x;s Journal of Research in Biology (2013) 3(4): 977-983

Anbalagan et al., 2013 Table 2. Diversity indices for Odonata in vegetable and rice fields from 2005 to 2008 Sl. No. 1 2 3 4 5 6

2005 Vegetable Rice Fields Fields 21 31

Diversity Indices Species richness (S) Total no. of individuals Shannon-Wiener Diversity Index (H) Simpson 1-D Evenness Jaccard Similarity Index

2006 Vegetable Fields 31

Rice Fields 21

2007 Vegetable Fields 31

Rice Fields 21

2008 Vegetable Fields 31

1464

3656

1368

3277

1622

3670

1703

4167

2.84

3.221

2.828

3.3

2.871

3.328

2.847

3.308

0.9326 0.815

0.9508 0.8082

0.9288 0.8056

0.9584 0.8749

0.9358 0.8409

0.9601 0.8991

0.934 0.8208

0.9591 0.8817

0.660

Table 3. Mateorological data for the years from 2005 to 2008 Year

Mean Maximum temperature (oC)

2005 2006 2007 2008

33.6 33.8 33.3 33.7

Mean minimum temperature (oC)

Mean Relative Humidity (%) @ 0830/1730 hrs IST 66.8-75.6 64.3-75.9 67.1-75.7 64.3-75.3

24.8 24.6 24.5 24.8

Mean Total Rainfall (mm)

Several

Meteorological data:

199.8 123.9 106.9 150.2 investigators

Total number of rainy days (2.5mm and above) 73 67 68 63 have

reported

that

The meteorological data is given in the table 3.

dragonflies and damselflies are very common in rice

Mean maximum and minimum yearly temperatures were

agroecosystems. Kandibane et al., (2003) have recorded

low in 2007 compared to other three years. Also the

12 species of Odonata under three families in rice fields

relative humidity was high in the year 2007.

of Madura. In the present work the number of species and families recorded in rice fields were high compared to the results of Kandibane et al., (2003, 2005). Among

DISCUSSION Present study reports the odonata diversity in

the various species, Pantala flavescens, a migratory

vegetable and rice agroecosystems. Odonates are

species, was abundant in numbers. The damselfly

predaceous insects and they are important biocontrol

Ceriagrion coromandelianum was abundant in both

agents of agricultural pests and vector mosquitoes.

vegetable

In the present study families Libellulidae in Anisoptera

Agriocnemis femina femina was also found to be

and Coenagrionidae in Zygoptera were found to be more

abundant. Kandibane et al., (2003) have reported that

diverse families in terms of the number of species.

A. femina was more abundant in rice ecosystems.

fields

and rice

fields.

rice

field,

Similar findings were already reported by some

The diversity and distribution of insects may be

investigators. Ghahari et al.,(2009) have reported that

influenced by type of ecosystems and climate. In the

families Libellulidae and Coenagrionidae were dominant

present study the species richness, total abundance and

in terms of number of species in rice fields in Iran.

diversity of Odonata were high in vegetable ecosystems

Kumar

family

compared to rice ecosystem. Higher evenness values

Libellulidae was represented by high number of species

were recorded in vegetable fields than rice fields during

(18 species) among a total collection of 42 species from

2006, 2007 and 2008.

Sahstradhara,

were

evenness of a community increases, the Shannon index

published by Prasad (2002), Kumar (2002) and

also increases. In the present study the Shannon index

Vashishth et al., (2002).

was higher in vegetable fields than rice fields. This was

and

Mitra

Dehra

(1998)

Dun.

reported

Similar

that

reports

Journal of Research in Biology (2013) 3(4): 977-983

When the richness and the

981

Anbalagan et al., 2013 due to the higher species richness and evenness in

Fraser FC. 1933. The Fauna of British-India Including

vegetable fields. The dominance of species was found to

Ceylon and Burma, Odonata.vol.1, Taylor and Francis

be lower in vegetable crops compared to rice fields.

Ltd., London.

Hence the Simpson„s index of diversity (1-D) was higher in vegetable crops and it clearly explained that species distribution in vegetable crops was equal. Besides the type of crop, the climatic factors

Fraser FC. 1934. The Fauna of British-India Including Ceylon and Burma, Odonata. vol. II. Taylor and Francis Ltd., London.

such as rainfall, atmospheric temperature and humidity

Fraser FC. 1936. The Fauna of British-India Including

also affect the insect diversity. The average annual

Ceylon and Burma, Odonata. vol. III. Taylor and Francis

temperature was the lowest in the year 2007. This lowest

Ltd., London.

average temperature in 2007 coincided with the maximum insect diversity in both rice and vegetable crops. Brinjal and okra plants grow taller with branches and provide suitable microclimate and resting place for perching adult Odonata. Vegetable fields also harbour

Ghahari H, Tabari M, Sakenin H, Ostovan H and Imani S. 2009. “Odonata (Insecta) from Northern Iran, with comments on their presence in rice fields,” Mun. Ent. Zool, 4(1): 148-154.

variety of small insects, which are the main prey of

Gunathilagaraj K, Soundarajan RP, Chitra N and

Odonates. Latif et al., (2009) have reported 20 species

Swamiappan M. 1999. “Odonata in the rice fields of

of pest insects and 10 families of predaceous insects in

Coimbatore”, Zoo`s Print J., 14(6): 43-44.

brinjal field. Hence the presence of variety of prey insects might be the reason for higher odonata diversity in vegetable fields.

Kandibane M, Mahadevan NR and Gunathilagaraj K. 2003. “Odonata of irrigated rice ecosystem of Madurai, Tamil Nadu,” Zoo’s Print J., 18: 1155-1156.

CONCLUSION

Kandibane M, Raguraman S and

Ganapathy N.

It is concluded that dragonfly and damselfly

2005. “Relative abundance and diversity of Odonata in

diversity was influenced by type of crop because

an irrigated rice field of Madurai, Tamilnadu,” Zoo’s

vegetable ecosystem supported more taxa of Odonates

Print J., 20(11): 2051-2052.

than rice field.

Kumar A. 2002. “Odonata Diversity in Jharkhand State

ACKNOWLEDGMENTS The authors thank Entomology Research Institute for financial support

with Special Reference to Niche Specialization in their Larva Forms,” In: Current Trends in Odonatology, Kumar, A., (Ed.). Daya Publishing House, New Delhi, 297-314.

REFERENCES

Kumar A and Mitra A. 1998. “Odonata diversity at

Corbet PS. 1999. Dragonflies: Behavior and Ecology,

Sahastredhara (Sulphur springs), Dehra Dun, India,” with

Cornell University Press, Ithaca, New York, 829.

notes on their habitat ecology. Fraseria, 5(1/2): 37-45.

Emiliyamma

2005. “On the odonata (Insects)

Latif MA, Rahman MM, Islam MR and Nuruddin

fauna of Kottayam District, Kerala, India”. Zoos Print J.,

MM. 2009. “Survey of arthropod biodiversity in the

20(12): 2108-2110.

brinjal field,” J. Entomol., 6(1): 28-34.

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Anbalagan et al., 2013 Mandal SK, Ghosh A, Bhattacharjee I and Chandra G. 2008.“Biocontrol efficiency of odonate nymphs against

larvae

the

mosquit o,”

Culex

quinquefasciatus Say, 1823. Acta Tropica,106(2):109114. Prasad M. 2002. “Odonata Diversity in Western Himalaya, India,” In: Current Trends in Odonatology, A. Kumar, (Ed.). Daya Publishing House, Delhi, 221-254. Prasad M and Varshney RK. 1995. “ A checklist of odonata of india including data on larval studies,” Oriental Insects, 29(1): 385-428. Spencer M, Blaustein L, Schwartz SS and Cohen JE. 1999. “Species richness and the proportion of predatory animal

species

temporary

freshwater

pools:

relationships with habitat size and permanence,” Ecol. Lett., 2(3): 157-166. Sharma G. 2010. “Studies on odonata and lepidoptera (Insecta: Arthropoda) fauna of Mount abu, Rajasthan, India,” Hexapoda, 17: 136-141. Subramanian KA. 2009. Dragonflies of India: A Field Guide. Vigyan Prasar, New Delhi, India. Tsuda S. 1991. A Distributional List of World Odonata. Privately Published, Osaka, Japan, pages: 362. Vashishth N, Joshi PC and Singh A. 2002. “Odonata community dynamics in Rajaji National Park, India,” Fraseria, 7: 21-25. Submit your articles online at www.jresearchbiology.com Advantages

Easy online submission Complete Peer review Affordable Charges Quick processing Extensive indexing You retain your copyright submit@jresearchbiology.com www.jresearchbiology.com/Submit.php.

Journal of Research in Biology (2013) 3(4): 977-983

983

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Hepatic enzyme markers and proteins in serum and some selected tissues in Clarias gariepinus from swamp around Kokori-Erhoike oil field, Nigeria Authors: Osioma E1*, Akanji MA1 and Arise RO1.

ABSTRACT: This study determines changes in some biochemical parameters in serum and tissues of Clarias gariepinus obtained from fish natural habitat in the oil exploration environs of Kokori-Erhoike in Delta State, Nigeria. Sampling sites include Ethiope River (Eku axis, reference Site A); Erhoike swamp (Site B) and Erhoike fish pond (Site C). However, Sites B and C are located in the oil exploration region of Erhoike. Clarias gariepinus (n=8) were collected from each site and used for the study. Levels of total proteins, albumin, haemoglobin as well as the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase were determined in serum, gill, liver, brain and muscle tissues. Results showed that total protein concentrations were comparable (p>0.05) in serum. Albumin concentrations of fish from Site B and C were lower (p<0.05) as compared with that of Institution: site A in gill, muscle and brain tissues. Haemoglobin concentration was significantly 1. Department of lower (p<0.05) in fish from Site A as compared with that of Sites B and C. Results also Biochemistry, Faculty of Science, University of Ilorin, indicated that total protein and albumin concentrations were significantly (p<0.05) Nigeria. higher in gill, muscle and brain tissues of Clarias gariepinus from Site A as compared with fish tissues from Sites B and C. Hepatic enzymes (ALT and AST) and ALP activities were elevated (p<0.05) in serum, gill, brain and liver of fish from Sites B and C as compared with that of Site A. The observed biochemical changes in fish from Sites B and C could have resulted from contaminants arising from the oil exploration activities in Site B and the presence of organic/inorganic contaminants in Site C due to the presence of fish feeds. These biochemical alterations show that the fish were under stress in their natural habitat. These biomarkers could be employed in the environmental monitoring of crude oil pollution as well as early warning signs of the adverse effects of environmental pollution. Corresponding author: Osioma E.

Keywords: Fish, Kokori-Erhoike, Clarias gariepinus, Albumin, Alkaline phosphatase, Haemoglobin.

Email:

Article Citation: Osioma E, Akanji MA and Arise RO. Hepatic enzyme markers and proteins in serum and some selected tissues in Clarias gariepinus from swamp around Kokori-Erhoike oil field, Nigeria. Journal of Research in Biology (2013) 3(4): 984-992

Web Address:

Dates:

http://jresearchbiology.com/ documents/RA0343.pdf.

Received: 23 Mar 2013

Accepted: 23 May 2013

Published: 13 June 2013

Journal of Research in Biology An International Scientific Research Journal

984-992 | JRB | 2013 | Vol 3 | No 4

www.jresearchbiology.com

Osioma et al., 2013 group from α-amino acid to α-keto acid. AST and ALT

INTRODUCTION Over the years, the extent of oil exploration

are biological responses of severe hepatic injury and

activities and its related environmental effects has been

their bioassay can serve as a diagnostic tool for

on the increase, (Tolulope, 2004). Some quantities of

estimating necrosis of the liver cells. (Cappo et al.,

petroleum and its products may be released into the

2002). The determination of ALT and AST activities has

environment during oil exploration activities due to

been applied in fish research to indicate bacteria, viral

operational, accidental, transportation or other means.

and parasitic infection, intoxications and water pollution

Apart from contaminating the flesh of commercially

(Bucher and Hofer, 1990). Alkaline phosphatase

valuable fish, crude oil compromise fish hatcheries in

comprises group of enzymes which is responsible for

coastal waters (Leighton, 1991) and its products are the

hydrolyzing phosphoric ester bonds present in organic

most relevant to aquatic ecotoxicology (Pacheco and

compounds at an alkaline pH (Akcakaya et al., 2007).

Santos, 2001). In Nigeria, crude oil was discovered at

The enzyme (ALP) has been reported to be a marker

Oloibiri in 1959 (Akpofure et al., 2000) and exploration

enzyme for the plasma membrane and endoplasmic

activities has been carried out in Kokori- Erhoike

recticulum (Akanji et al., 1993).

environment for over 30 years (Emoyan, 2009).

Fish species are excellent subjects for the study

The cause-effect relationship and result of

of various effects of contaminants (El-Shehami et al.,

xenobiotic pollution in an ecosystem can be assessed

2007) and African catfish (Clarias gariepinus) has been

through the analysis of biochemical alterations on

used in fundamental research and toxicological studies

organisms

(Nguyen and Janssen, 2002)

inhabiting

that

environment.

These

biochemical alterations may be sensitive and specific as

A significant body of research has investigated

early indicators of aquatic pollution (Norris et al., 2000;

the effects of crude oil (or its derivatives) and refinery

Strinac and Braunbeck, 2000)

effluents on fish health. The investigators include:

Protein plays a vital role in the physiology of

Yarbrough et al., 1976; Kuehn et al., 1995; Sunmonu

living organisms and its metabolism according to Adams

and Oloyede, 2006; Wegwu and Omeodu, 2010;

et al., (1990) provides information on the general energy

Mahmoud et al., 2011 and Nwaogu et al., 2011.

mobilization of an animal and show relationship with

However, there is dearth of information on the effect of

effects of contaminants in these organisms. The

crude oil exploration activities on African catfish

concentration of plasma albumin is a useful index of the

(Clarias gariepinus) obtained from swamps (fish natural

state of protein repletion and it makes the major

habitat) around Kokori- Erhoike oil field located in Delta

contribution to plasma sulphydryl groups which can

State, Nigeria. This information deficiency prompted this

function as a chain breaking antioxidant (Halliwell,

study. Therefore, the aim of this research is to determine

1988). Haemoglobin contained in the red blood cells

changes of some biochemical parameters in serum and

which serve as the oxygen carrier in blood has been

tissues

employed in assessing the health of fish and monitoring

Clarias gariepinus obtained from swamps around Kokori

stress response of several environmental contaminants

- Erhoike petroleum flow station in Delta State, Nigeria.

(gill,

liver,

brain

and

muscle)

including petroleum hydrocarbons (Soivio and Oikari., 1976; Gabriel et al., 2007). Alanine aminotransferase (ALT) and aspartate

MATERIALS AND METHODS Sampling sites:

aminotransferase (AST) catalyze the transfer of α-amino 985

Journal of Research in Biology (2013) 3(4): 984-992

Osioma et al., 2013 This study was carried out in Ethiope East Local

Kokori-Erhoike petroleum flow station where oil

Government Area of Delta State, Nigeria. Experimental

exploration activities have been on for more than 35

areas have been previously described by Aries et al.,

years. This area has a number of oil wells and flow

2013 and are represented in Figure 1 below.

stations. The aquatic ecosystem in the area is constituted

Site A (reference site) is the Eku axis of the

by non-tidal freshwater swampy forest characteristics of

Ethiope River, Delta State. There is no presence of oil

those found within the freshwater survey zone of the

facilities/operations or any industry located along the

Niger Delta. Site C is a natural fish pond located within

Ethiope River from its source, Umuaja, about 22km to

Kokori-Erhoike environment. The main uses of water in

the Eku axis. The upper axis of the Ethiope River has

the catchments include domestic, recreational (e.g.

been reported to be relatively unpolluted (Ikomi et al.,

swimming) and fishing. Their major occupation includes

2005; Agbaire and Obi, 2009; Aries et al., 2013). This

farming (cassava, yam, Okro etc.), fishing and petty

qualifies the Eku axis of Ethiope River as a reference site

trading on food stuff.

for this study. Site B is the swampy environment of

Fig 1. Map of Ethiope East showing the location sampling sites Source: Ministry of lands, Surveys and urban Development, Asaba. (2008) Journal of Research in Biology (2013) 3(4): 984-992

986

Osioma et al., 2013 Duncan’s Multiple Range Test (DMRT) at 5%

Fish Eight African catfish (Clarias gariepinus) were collected from each site in October, 2011. At site A (Eku

probability level. All statistical analysis was performed using SPSS version 16.

River) and site B (Erhoike swamp), fish were caught with the help of professional local fishermen and at site C (Erhoike Fish pond) fish net was used to catch the fish.

RESULTS Results (Table 1) show that gill, muscle and brain total protein concentrations were significantly

Preparation of Serum About 1.5ml blood was taken by caudal arterial

higher (p<0.05) in fish from Site A as compared with

puncture from each fish into a sterilized plain tube.

that of Site B and C. Total protein concentration in serum

Blood was allowed to clot for about 5min dislodged and

and liver of Clarias gariepinus from all sites were

centrifuged at x15,000g for 15min to obtain the serum,

comparable at p>0.05. The data also indicated significant

which was stored frozen at – 20°C until analyzed.

reduction

Preparation of Tissue Homogenate

concentration of Clarias gariepinus from Sites B and C

(p<0.05) in

gill

and muscle albumin

The fish were dissected and the gill, liver, brain

as compared with that of Site A. Levels of serum and

and muscle tissues were quickly removed. The tissues

liver albumin were comparable in fish from all sites.

(gill, liver, brain and muscles) were washed in cold

Results in Table 1 also revealed that fish from Sites B

saline (0.9% NaCl) solution several times and then 1g of

and C has elevated (p<0.05) haemoglobin concentration

wet tissue was homogenized in 9 ml of the physiological

compared with that of Site A.

solution (normal saline). The resulting homogenate was centrifuged at 5000g for 20min. The supernatant was decanted and used for further biochemical analysis. Biochemical Investigations The concentrations of total protein and albumin were determined in serum and tissues (gill, liver, muscle

Table 1: Levels of total protein, albumin and haemoglobin concentrations in serum and tissues (gill, liver, muscle and brain) of Clarias gariepinus from swamps around Kokori-Erhoike Petroleum Flow Station in Delta State, Nigeria SAMPLING SITES A B C Total protein concentration (g/dl)

and brain) employing the methods of Doumas et al., 1981

and

Doumas

al.,

1971

respectively.

Haemoglobin level was estimated by the method of Tietz (1976), while the method of Roy (1970) was used to determine the activities of alkaline phosphatase. Alanine aminotransferase and aspartate aminotransferase were analyzed using the method of Reitman and Frankel (1957). All assays were carried out with the aid of commercially

available

kits

supplied

TECO

Diagnostics, Anahem, USA and Randox Laboratories, Ardmore, United Kingdom. Statistics Analysis of Variance (ANOVA) was used to analyse data obtained from the various biochemical investigations. Group means were compared by the 987

Serum (n=8) Gill (n=8) Liver (n=8) Muscle (n=8) Brain (n=8)

9.31±0.53a 2.82±0.05a 2.81±0.37a 5.28±0.32a 4.17±0.17a

9.30±0.31a 2.46±0.24b 2.91±0.15a 4.29±0.10b 3.72±0.13b

9.29±0.33a 2.43±0.03b 2.98±0.40a 4.08±0.40b 3.96±0.12c

Serum (n=8) Gill (n=8) Liver (n=8) Muscle (n=8) Brain (n=8)

Albumin concentration (g/dl) 7.25±0.32a 7.01±0.01a 6.96±0.13a 2.48±0.36a 2.03±0.16b 2.08±0.24b a a 2.52±0.43 2.51±0.26 2.69±0.18a a b 4.09±0.17 2.09±0.42 2.55±0.12b 3.78±0.22a 2.25±0.11b 3.68±0.10a Haemoglobin concentration (g/dl)

Blood (n=8)

22.24±1.57a

27.03±1.65b

27.29±1.65b

Values are given as Mean ± SD. Means not sharing a common superscript letter on a given row differ significantly at p<0.05. A= Ethiope River (Eku axis); B= Erhoike swamp; C= Erhoike fish pond. Journal of Research in Biology (2013) 3(4): 984-992

Osioma et al., 2013 Increased aminotransferase

(p<0.05) in

activities

serum

and

alanine

liver

Clarias gariepinus from Sites B and C were observed as compared with that of Site A (Table 2). Similar trend was observed for the activity of liver aspartate

Table 2: Changes in the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) in serum and tissues (liver, gill and brain) of Clarias gariepinus collected from swamps around Kokori-Erhoike Petroleum Flow Station in Delta State, Nigeria SAMPLING SITES A (n=8) B (n=8) C (n=8) Alanine aminotransferase (IU/L) 43.86±2.79a 48.28±0.34b 49.19±0.65 b 25.10±0.80a 45.90±1.63b 42.38±1.82c Aspartate aminotransferase (IU/L) 53.75±8.84 a 54.34±2.88 a 54.25±3.15 a 52.31±0.79a 69.70±1.63b 65.94±1.82c Alkaline phosphatase (IU/L) 32.86±3.14 a 46.22±3.05 b 45.76±3.49 b 43.01±1.48a 52.67±1.79b 53.83±2.91b 29.73±3.73a 36.79±1.39b 32.74±5.36a 52.94±2.36a 57.34±1.21b 60.37±2.25c

aminotransferase. The table also showed that the activity of aspartate aminotransferase in serum of fish from all sites (A, B and C) were comparable (p>0.05), although, fish from Site A had relatively lower aspartate aminotransferase activity. Results (Table 2) also showed that the activity of alkaline phosphatase in (serum, liver, gill and brain) of Clarias gariepinus from Sites (B and C) were significantly (p<0.05) higher as compared with that of Site A. DISCUSSION Biochemical markers of pollution are considered

Serum Liver Serum Liver Serum Liver Gill Brain

Values are expressed as Mean± SD. Means not sharing a common superscript letter on a given row differ significantly at p<0.05. A= Ethiope River (Eku axis); B= Erhoike swamp; C= Erhoike fish pond.

indicators employed in fish toxicity tests and for field monitoring of aquatic contamination. They established

contaminated the aquatic environment and petroleum

contact of the sample with definite groups of chemical

hydrocarbon can act as a mediator in free radical

compounds and clarify their metabolic fate. Biochemical

generation in fish (Achuba and Osakwe, 2003). During

investigations allow cause-effect relationship to be

stress conditions, fishes need more energy to detoxify the

established at an early stage of pollution and these

toxicants and to overcome stress, thus carbohydrate

sensitive and predictive diagnostic tools (Biomarkers) for

reserve is depleted to meet energy demand (Nelson and

assessing animal exposure and toxic effects of chemical

Cox, 2005; Sudhanshu and Ajay, 2009). Since fish have

contaminants are needed as aquatic environmental

a very little amount of carbohydrate, the next alternative

contamination assessment indicators.

source of energy is protein to meet the increased energy

The total protein concentration in serum, gill,

demand occasioned by a pollutant.

liver, muscle and brain of Clarias gariepinus from Eku

The decrease in total protein level observed in

River is higher than the values obtained from fish

the gill, brain and muscle tissue could be to meet the

samples in Erhoike swamp and Erhoike fish pond. The

higher energy demands for metabolic purposes due to the

differences were significant (p<0.05) in the gill, brain

presence

and muscle tissues.

environment and could also be related to impaired food

Proteins play a vital role in the physiology of living organisms and provide information on the general

petroleum

hydrocarbon

Erhoike

intake, increased energy cost of homeostasis, tissue repair and detoxification mechanism during stress.

energy mobilization of an animal and show relationship

Albumin is the most soluble and most electrically

with the effect of contamination in these organisms

mobile of all the major serum protein components and it

(Adams et al., 1990). The over 30 years of petroleum

is synthesized entirely by the hepatic parenchymal cells.

exploration activities in Erhoike vicinity could have

Lower albumin concentration was observed in the serum,

Journal of Research in Biology (2013) 3(4): 984-992

988

Osioma et al., 2013 gill, liver, muscle and brain tissues of Clarias gariepinus

Alanine

aminotransferase

and

aspartate

from Erhoike swamp and Erhoike fish pond compared

aminotransferase (ALT and AST) are enzymes directly

with corresponding albumin level of Clarias gariepinus

associated with the conversion of amino acids to keto

from Eku River (control site). The utilization of proteins

acids. Apart from being considered to be important in

as an alternative source of energy by fish in stress

assessing the state of the liver and some other organs

condition could have accounted for the reduced albumin

Verma et al., (1981), transamination of the same

level in fish from Erhoike swamp and Erhoike fish pond.

represents one of the main pathways for synthesis and

Albumin has also been regarded as an antioxidant

deamination of amino acid, thereby allowing interplay

molecule. It reacts with and neutralizes peroxyl radicals

between carbohydrate and protein metabolism during the

(Stocker and Frei, 1991) and it is considered as a

fluctuating energy demands of the organisms in various

sacrificial molecule that prevents damage when it acts as

adaptive situations. ALT and AST activities are direct

an antioxidant because albumin is destroyed in the

indicators of intense hepatic damage, thus their bioassay

process (Halliwell, 1988). Therefore, the observed

can assist as a diagnostic tool for determining necrosis of

reduction of albumin concentration in African Catfish

the liver cells (Whitehead et al., 1999; Cappo et al.,

from the oil exploration areas may be linked to its

2002). Ugwu et al. (2008) concluded that AST enzyme

participation as an antioxidant molecule to quench free

activity in Heterobranchus bidorsalis adults could be

radical reactions in other to mitigate the impact of

used as biomarker for monitoring crude oil pollution in

oxidative stress or its utilization as a source of energy by

Nigeria.

the fish in stress condition.

C om p a r e d

wi t h

th e

c on t r ol

(i . e .

This study showed that Clarias gariepinus from

Clarias gariepinus from Ethiope River, Eku axis) the

Erhoike swamp and Erhoike fish pond have higher levels

activities of ALT and AST in serum and liver of

(p<0.05) of haemoglobin as compared with haemoglobin

Clarias gariepinus from Erhioke swamp and Erhoike

concentration of Clarias gariepinus from the control site

fish pond were higher. Such increase of ALT and AST

(Eku River). Elevated levels of haemoglobin observed in

may be partly due to hepatic damage resulting from

African catfish from Erhoike swamp and fish pond could

petroleum pollution (in case of Erhoike swamp) or

be as a result of stress induced by the presence of crude

organic/inorganic contaminants (present in Erhoike fish

oil and other contaminants (as in the case of the fish

pond, Arise et al., 2013) â&#x20AC;&#x201C;induced oxidative insults on

pond) that leads to environmental hypoxia as a result of

the hepatocytes. In addition, increased protein catabolism

chronic exposure to the contaminants and anaerobic

might be responsible for the elevation of these

condition

transaminases. These results agree with the findings of

which

lead

increase

haemoglobin

concentration as a compensatory mechanism

for

increased oxygen demand. This result corroborates with

Ayalogu et al., (2001); Orisakwe et al., (2005). In this study, serum, liver, gill and brain alkaline

the findings of Mdegela et al., (2010) who reported

ph osph a t a s e

a ct i vi t y

wer e

m ea sur ed

significant elevation of haemoglobin concentration in

Clarias gariepinus from the three sampling sites. Marked

fish from Mzumbe sewage water. Zaki et al., (2010) also

increase in ALP activity was recorded in the serum, liver,

reported significant increase in haemoglobin levels in

gill and brain tissues of Clarias gariepinus from Erhoike

Tilapia zilli exposed to acute lethal concentration dose of

swamp and Erhoike fish pond as compared with ALP

lead (Pb).

activity of Clarias gariepinus from the control site (Eku River). ALP together with ALT and AST provide an

989

Journal of Research in Biology (2013) 3(4): 984-992

Osioma et al., 2013 indication of the degree of inflammation as well as

Akanji MA, Olagoke OA and Oloyede OB. 1993.

possible causes of hepatocellular damage as well as

Effect of Chronic Consumption of metabisulphite on the

distortion of the plasma membrane and endoplasmic

integrity of rat liver cellular system. Toxicol., 81: 173 –

recticulum.

179.

Results of the research agree with the

findings of Yarbrough et al., (1976) and Ayalogu et al., (2001).

Akcakaya H, Aroymak A and Gokce S. 2007. A quantitative colorimetric method of measuring alkaline phosphatase activity in eukayotic cell membranes. Cell

CONCLUSION The study demonstrates that the level of

Biol Int., 31(2): 186 – 190.

contamination is enough to cause changes in the protein,

Akpofure EA, Efere ML and Ayawei P. 2000.

albumin and haemoglobin level with attendant effect on

Integrated grass root post-impact assessment of acute

the integrity of hepatocytes as evidence in the elevated

damaging effects of continuous oil spills in the Niger

activities as seen in liver marker enzymes and alkaline

Delta. A paper report on oil spillage in the Niger Delta.

phosphatase activity of Clarias gariepinus from Erhoike

Source:

swamp and Erhoike fish pond. The above biochemical changes showed that the fish were under stress in their natural habitat (Erhoike swamp) thus; these markers

http://www.waado.org/environment/petrolpolution/ oilspill/OilSpills_AdverseEffects.html.

could be employed in the environmental monitoring of

Arise RO, Osioma E and Akanji MA. 2013.

crude oil pollutant and their associated metabolic

Assessment of water quality parameters from swamps

changes as early warning signs of adverse effects of

around Kokori-Erhoike petroleum flow station in Delta

environmental pollution.

State, Nigeria. Der Chemica Sinica., 4(1): 155-161. Ayalogu OE, Igboh NM and Dede EB. 2001.

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