PHARMACOLOGICAL AND CLINICAL EVALUATION OF THE ANTHELMINTIC ACTIVITY OF ALBIZIA ANTHELMINTICA BROGN, MAERUA EDULIS DE WOLF AND MAERUA SUBCORD AT A DE WOLF PLANT EXTRACTS IN SHEEP AND MICE If DANIEL WAWERU GAKUYA, BVM, MSc. UNIVERSITY 5 F n\ ir#w gABEIL U1RAJLX A thesis submitted in fulfillment of the requirements of Doctor of Philosophy Degree in Veterinary Clinical Studies Faculty of Veterinary Medicine University of Nairobi 2001 DECLARATION This thesis is my original work and has not been presented for a degree in any other University. Signed T** NniAUJM- Date 4 ^ 2 ^ .1s>ol. DANIEL WAWERU GAKUYA, BVM, MSc I his thesis has been submitted for examination with our approval as University supervisors. _________ Date c 3 - C f l2* PROF. P.M.F. MBITH1, BVM, MSc., MVSc., PhD \ _______ Date L z t S ^ ■ i - e o l . PROF. T.E. MAITHO, BVM. MSc., PhD — U J t$ A A /V ) <o>*_________ Date__ PROF. N.K.R MUSIMBA, BSc., MSc., PhD Department of Clinical Studies, Faculty of Veterinary Medicine, College of Agriculture and Veterinary Sciences, University of Nairobi, KENYA. li DEDICATION TO MV LOVlNQ PARENTS, THE LATE JAMES QAKUVA AND MUM MARVtfANJIKU ill TABLE OF CONTENTS PAGE DECLARATION........................................................................................................ii DEDICATION...........................................................................................................iii TABLE OF CONTENTS?^.. C ..................................................................... iv LIST OF TABLES................................................................................................. viii LIST OF FIGURES................................................................................................ix LIST OF APPENDICES...........................................................................................x ACKNOW LEDGEM ENT.................................................................................... xii ABSTRACT............................................................................................................ xiv CHAPTER 1. INTRODUCTION AND OBJECTIVES....................................1 1.0. 1.TRODUCTION.............................................................................................. 1 1.1.OBJECTIVES....................................................................................................... 4 CHAPTER 2. LITERATURE REVIEW .............................................................. 5 2.0. Helminthoses in Kenya....................................................................................... 5 2.0. 1. Economic importance of helminthoses............................................5 2.0. 2. Helminth control methods................................................................ 6 2.0. 2.1..Anthelmintics.................................................................................. 6 2.0. 2.2. Nutrition..........................................................................................6 2.0. 2.3. Management practices................................................................... 6 2.0. 2.4. Grazing management..................................................................... 7 2.0. 2.5. Helminth resistant breeds.............................................................. 8 2.0. 2.6. Destruction of intermediate host...................................................8 2.0. 2.7.Helminth vaccines...........................................................................8 2.0. 3. Constraints to helminth control........................................................8 2.0. 3.1. Finance.............................................................................................8 2.0. 3.2. Anthelmintic resistance................................................................. 9 2.0. 3.2.1. Causes of anthelmintic resistance.............................................10 2.1. Background of the usage of medicinal plants..................................................11 2.1.1. Development of Ethnoveterinarymedicine..................................................11 2.1.2. Plant anthelmintics.........................................................................................13 2.1.2.1. Plant that have been used to treat livestock helminthoses.................... 13 2.2. Methods used to identify plants of medicinal value and pharmacological parameter............................................................................................................. 15 iv 2.2.1. Survey questionnaire......................................................................................15 2.2.2. Methods for extraction....................................................................................16 2.2.3. Detecting bioactivity in plant material using brine shrimp lethality test.....................................................................................................17 2.2.4. Toxicity tests and determination of LDso......................................................18 2.2.5. Determination of the percentage faecal egg counts reduction................. 20 2.2.6. Separation and identification of the pharmacological active ingredient in the plant anthelmintic..............................................................32 2.2.7. Limitation of plant anthelmintics..................................................................33 2.2.7.1. Ethnodiagnosis...............................................................................33 2.2.7.2. Toxicity...........................................................................................34 2.2.7.3. Availability..................................................................................... 34 2.2.7.4. Sustainability.................................................................................. 35 2.2.7.5. Traditional beliefs.......................................................................... 35 CHAPTER 3 IDENTIFICATION OF MEDICINAL PLANTS USING PARTICIPATORY RURAL APPRAISAL TOOLS........................................37 3.0 Introduction........................................................................................................37 3.1. Materials and methods...................................................................................... 39 3.2. Results...............................................................................................................41 3.3. Discussion.......................................................................................................... 47 CHAPTER 4 DETERMINATION OF THE EFFICACY OF CRUDE WATER EXTRACTS OF ALBIZIA ANTHELMINTICA BROGN AND MAERUA SPECIES ON GASTROINTESTINAL NEMATODES IN SHEEP....................................................................................................................... 49 4.0. Introduction........................................................................................................ 49 4.1. Materials and methods...................................................................................... 51 4.1.0. Plant material...................................................................................... 41 4.1.1. Animals............................................................................................... 52 4.1.2. Experimental design...........................................................................52 4.1.3. Statistical analysis.............................................................................53 4.2. Results and discusion.........................................................................................53 CHAPTER 5 DETERMINATION OF THE BIOACTIVITY OF PLANT EXTRACTS USING THE BRINE SHRIMP LETHALITY TEST.............. 57 5.0. Introduction.........................................................................................................57 5.1. Materials and methods.......................................................................................58 5.1.1. Acquisation and preparation of plant material............................... 58 5.1.2. Extraction of active ingredient from plant material........................58 5.1.3. Hatching of the brine shrimp............................................................. 60 5.1.4. Bioassay.............................................................................................. 61 5.2. Results and Discussion......................................................................................64 v CHAPTER 6 EVALUATION OF THE EFFICACY OF PLANTS EXTRACTS AGAINST NEMATODE HEL1GMOSOMOIDES POLYGYRUS INFECTIONS IN M ICE........................................................... 73 6.0. Introduction.........................................................................................................73 6.1. Materials and methods...................................................................................... 74 6.1.0. Experimental design........................................................................ 74 6.1.1 .Treatment regime................................................................................ 75 6.1.2. Parasitological techniques...............................................................77 6.1.2.1 .Faecal egg count concentration (EPG) determination................ 95 6.1.2.2. Accurate determination of total worm burden.............................78 6.1.3.Statistical analysis.............................................................................. 82 6.2. R esults...............................................................................................................