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Organochlorine Pesticides Residues in Fish and Sediment from Lake Naivasha ' ORGANOCHLORINE PESTICIDES RESIDUES IN FISH AND SEDIMENT FROM LAKE NAIVASHA ' Japhet Mworia M Anampiu BSc in Biochemistry/Zoology, University of Nairobi A project Research Report submitted in partial fulfillment for the Degree of Master of Veterinary Public Health, in the University of Nairobi University ot NAIROBI Library IKIPUI 0460013 6 Department of Public Health, Pharmacology and Toxicology, Faculty of Veterinary Medicine, College of Agriculture and Veterinary Science, University of Nairobi 2011 DECLARATION This report is my original work and has not been presented for a degree in any other university. 1 1 ] 1 J.M. M'ANAMPIU. \)i DateJ..|s..i....^:.^..!..\.............. This report has been submitted for examination with our approval as university supervisors. DR. L. W .KANJA (BSc, MSc, PhD). ate. JL*f: University of Nairobi Department of Public Health, Pharmacology and Toxicology Department of Public Health, Pharmacology and Toxicology o/( PROF. F.M. NJERUH (B.V.M, MSc, a University of Nairobi Department of Public Health, Pharmacology and Toxicology ii D EDICATION This report is dedicated to my wife Lydia, children and my parents iii ACKNOWLEDGEMENTS I am sincerely grateful to my university supervisors Dr.Laetitia.W.Kanja, Dr. James Mbaria and Prof.Francis.M.Njeruh for their guidance and assistance through out the period of this study. My thanks also go to the Naivasha District fisheries Office staff for their help during samples collection from Lake Naivasha, especially Mr.Rodrick Kundu who was in charge of the station. A good deal of appreciation goes to the lake Naivasha Riparian Association and the Kenya marine and fisheries institute (KMFRI) Lake Naivasha station for availing information that was helpful to me during my research work. Mr.Francis.K.Gitau, Department of Public Health, Pharmacology and Toxicology is highly acknowledged for his technical assistance in the analysis of the samples and to the entire staff of the Pharmacology and Toxicology Laboratory for their assistance. I would also like to acknowledge Dorcas chege for her help in statistical analysis of data. I sincerely thank my entire family for encouragement and patience throughout the entire course of my study. Finally I profoundly thank Prof.Francis.M.Njeruh who was the chairman of the Department of Public Health, Parmacology and Toxicology from the bottom my heart for the co-operation accorded and for allowing me to carry out research in the department. The lecturers generously offered their extra time and showed patience with me during the course of the study and for this I thank them sincerely. IV TABLE OF CONTENTS Page DECLARATION jj DEDICATION jjj ACKNOWLEDGEMENTS iv LIST OF TABLES viii LIST OF APPENDICES ix ABBREVIATIONS x ABSTRACT xi CHAPTER ONE 1 1.0 INTRODUCTION 1 CHAPTER TWO 4 2.0 LITERATURE REVIEW 4 2.1 General Definition and Pesticides 4 2.2 Classification of organochlorine pesticides 8 2.2.1 DDT and Its Analogues 8 2.2.2 Cyclodienes 9 2.2.3 HexachloroCyclohexanes (HCH) 9 2.2.4 Toxaphene Group 10 2.3 Absorption and Accumulation of Organochlorine Pesticide in Fish 10 2.4 Metabolism and Excretion of Organochlorine Residues in Fish 11 2.4.1 Metabolism. 11 2.4.2 Excretion of Organochlorine Residues by Fish 12 2.5 Toxic Effects of Organochlorines in Fish 12 2.5.1 Acute toxicity *3 v 2.5.2 Chronic toxicity and sublethai exposure 14 2.6 Review of organochlorine pesticides residues in fish in the world 15 2.7 Review of studies on organochlorine pesticide residues in Kenyan fish 18 CHAPTER THREE 20 3.0 MATERIALS AND METHODS 20 3.1 Study site 20 3.2 Sample collection 22 3.2.1 Sample procedure 22 3.2.2. Species of fish sampled 22 3.3. METHODS 23 3.3.3 Washing of Concentrated Sulphuric acid with hexane 23 3.3.4 Preparation of pesticide standards 24 3.3.5 Gas liquid chromatograph (GLC) 24 3.3.6 Operating conditions 24 3.3.7_Detection limits 25 3.4 Extraction of Organochlorine residues 25 3.4.1 Clean-up 25 3.4.2 Acid clean up 26 3.4.3. Base clean up 26 3.4.4. Quantification of the pesticide residues 27 3.4.5 Confirmation 27 3.5. Toxicological Evaluation . 28 3.6. Statistical analysis • 28 CHAPTER FOUR 31 4.0 Results 31 4.1 Levels of organochlorine pesticides residues in Fish samples 31 vi 4.2 Levels of organochlorine pesticide residues in Sediment samples 32 4.3 Toxicological results 32 CHAPTER FIVE 38 5.0 Discussion 38 5.1 Organochlorines contaminants 39 5.1.1 Hexachlorocyclohexanes group 39 5.1.2 Cyclodiene group 40 5.4.3 DDT group 40 5.5 Maximum residue limit (MRL) and Acceptable Daily Intake (ADI) 41 5.6 Toxicological significance of the results 41 5.7 Comparison of health results with others on different studies 42 CHAPTER SIX 43 6.0 Conclusions and recommendations 43 6.1 Conclusions 43 6.2 Recommendations 43 REFERENCES 44 APPENDICES 48 vn LIST OF TABLES Table 1: Importation of different groups of pesticides into Kenya (1986-1995), Quantity in tonnes..............................................................................................................................4 Table 2: Legislation Relating to Pesticides in Kenya........................................................................... 6 Table 3: Test of Homogeneity of Variances................................................................................28 Table 4: Anova for organochlorine Pesticide residues differences between the fish species............................................................................................................................................... 29 Table 5: Anova for the organochlorine pesticide residues in liver, eggs, muscles and sediment.... 30 Table 6: The MRL and ADI for the pesticide residues detected. Values (FAO/WHO codex Alimentarius commission 1986)..........................................................................................................33 Table 7: Mean standard deviation (SD) and ranges of pesticide residue levels (mg/kg) in two species offish obtained from Lake Naivasha.........................................................................34 Table 8: Mean, Standard Deviation (SD) and ranges of pesticide residue levels (mg/kg) in the different sexes of the two fish species (tilapia and black bass) two sexes sampled...........................................35 Table 9: Mean, standard deviation (SD) and ranges of pesticide residue levels (mg/kg) in the liver, muscle and eggs of the two species (tilapia and black bass)................................................................36 Table 10: Mean and standard deviation (SD) of pesticide residue levels (mg/kg) in Lake Naivasha sediment Pesticide residue (mg/kg)..................................................................................................... 37 vm LIST OF APPENDICES APPENDIX 1: Form used during sample collection in Lake Naivasha............................48 APPENDIX 2: Pesticides residue levels in livers of black bass fish................................. 49 APPENDIX 3: Pesticide residue levels in eggs of black bass fish..................................... 50 APPENDIX 4: Pesticides residue levels in muscles of black bass fish...............................51 APPENDIX 5: Pesticide residue levels in muscles of tilapia fish....................................52 APPENDIX 6: Pesticide residue levels in eggs of tilapia fish...........................................53 APPENDIX 7: Pesticide residue levels in livers of tilapia fish .........................................54 APPENDIX 8: Pesticide residue levels in the lake sediment .......................................... 55 IX ABBREVIATIONS FAO Food and Agriculture Organization WHO World Health Organization KMFRI Kenya Marine and Fisheries Research Institute GLC Gas Liquid Chromatograph DDT 1,1,1 -trichloro-2, 2-di- (p-chlorophenyl) ethane HCH Hexachloro Cylohexane ANOVA Analysis of Variance MRL Maximum residue limit. ADI Acceptable daily intake LC50 Lethal concentration 50%. x ABSTRACT The main objective of this study was to identify and determine the levels of organochlorine pesticides residues in fish and sediment from Lake Naivasha and to evaluate the toxicological implications of their presence. A total of 82 fish samples and 24 samples of sediment were analyzed using a Packard gas liquid chromatograph fitted with a eiNi electron capture detector. The fish samples comprised of liver, eggs and muscle tissues. Out of 82 samples of fish analyzed 93.9% had detectable levels of at least one of the organochlorine compounds. Hexachlorocyclohexanes were the most frequently detected pesticides residues, a -HCH showing in 93.9% of the samples, 3-HCH in 60.9% of the samples and lindane in 6% of the samples. The cyclodienes were also detected with 50% of fish samples having heptachlor which had a mean of 0.004mg/kg, dieldrin was detected in 26 samples with a mean of 0.023 mg/kg, heptachlor epoxide in 25 samples with a mean of 0.0052mg/kg, aldrin was detected in only one liver sample of a female black bass and endrin was also found in only one liver sample of a male Tilapia fish. The 24 sediment samples analyzed, 62.5% showed presence of a -HCH with a mean of 0.003mg/kg, 37.5% samples had 3-HCH with a mean of 0.00315mg/kg, 12 samples had lindane with a mean of 0.00175mg/kg; Heptachlor epoxide was detected in only one sample with a value of 0.0012mg/kg, dieldrin was found in one sample with a value of 0.0033mg/kg and aldrin in one sample with a mean of 0.0025mg/kg. No DDT or its analogue was detected in fish and sediment samples. This showed that there has not been any recent use of the pesticide
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