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Icipelibrary ACC No BIO-PROSPECTING FOR INSECTICIDAL SCORPION VENOMS AND IDENTIFICATION OF BIOACTIVE TOXINS By John Bwire Ochola B.Sc. (Hon.),JKUAT A thesis submitted in partial fulfillment for the Degree of Master of Science in Chemistry in the Jomo K.enyatta University of Agriculture and Technology icipeLIBRARY ACC No .. J•• Cl ••::- ..U~ . .C .f:-1:: ............. 2005 CLASS NoI J:l.. ,5 . ~. 5.. : .~.~... Q.~};\ AUTHOR.0.~S'!~Q-. i .• : .: . ~ ........... TITLE. t2~it :: . r. .~~?.~~ .~~ : .~5: . ::-.. : .. : .. 11 DECLARATIONS The work described in this thesis was carried out at International Centre of Insect Physiology and Ecology (ICIPE). Information derived from published work is specifically acknowledged in the text and references appended. The experimental work described is entirely my own. This thesis is my original work and has not been presented for a degree in any other University. ------~~---------------------------- Date------&:1- !!J_~_t(p_ :i_ _______________ _ John Bwire Ochola This thesis has been submitted for examination with our approval as University Supervisors ' . - . ~~AJ~)-~ 21--11-2-005 1. Signature------- ~ - ~ -~ ~--- ~---~ -------------------- Date------------------------------------- Prof. Thuku G. Thiong'o . Chemistry Department Jomo Kenyatta University of Agriculture and Technology Nairobi, Kenya I.~ -:i-1-11-2ooS 2. Signature ------------~~-------------------------- Date------------------------------------- Dr. Wilber Lwande Behavioral and Chemical Ecology Department International Centre of Insect Physiology and Ecology Nairobi, Kenya. 3. Signature --------------------~----- Date--~_.. _ _l_L: __ ~_q_f?_____ _ Dr. Lucie M. Rogo BioNET-International Washington D.C., USA. 4. Signanrre -- = ---- -- ------~~-~.J:,;;-Nili~~~- Date-.?:Jt~-~ 1 --?_f_ _______________ _ ~ Department Kenyatta University Nairobi, Kenya. Ill DEDICATION To my wife,Jackline and our son; Nathan & my parents, Sabastian and Celementina; my sisters; my cousin Hilary IV ACKNOWLEDGEMENT Many people contributed to the success of my study in one way or another and I may not be able to list them all. However, I must mention some individuals and institutions without whose help and encouragement I may not have completed this thesis. I wish to thank Dr. Wilber Lwande, my supervisor at ICIPE for his academic, moral and material support during the entire period of my work I am equally grateful to Prof. Thuku Thiong'o, my University supervisor for his constant discussions and suggestions. I am equally indebted to Dr. Lucie Rogo for her encouragement, guidance, and invaluable advice to undertake the study. I am grateful to ICIPE Capacity building for the scholarship that enabled me to carry out my work under the Dissertation Research Internship Programme (DRJP). I am also grateful to Prof. Isaiah Ndiege for his contribution that gave me scientific direction and continued supervision of this research up to the end. The project was developed under the guidance of my promoter, Dr. Rafael Herrmann whom I sincerely appreciate for his tireless efforts. I further thank Dr. Scott Miller for supplying me with reference materials, Drs. Osir, Tsanuo K., Gikonyo N., Omulokoli E. and Okech M. for their valuable advice towards this work. I also recognize Dr. Prendini Lorenzo of AMNH for his valuable services in taxonomic identification of scorpions and Eric Schepers for the spectroscopic and amino acid analysis. My heartfelt gratitude goes to the entire staff and students of the ICIPE's B&CE Department, Biodiversity & Conservation subdivision and Bioprospecting Programme. I sincerely thank M. Koko, E . Nyandat, B. Nyambegah D. Mbuvi, R. Marubu and E . Ndenga for moral and technical assistance. A big thank you to my fellow students P. Mutungi, G. Ong'amo, V. Mahiva, E. Akhusama and N . Otieno for the morale and academic-sharing sessions. This work was done with full knowledge, permission and cooperation of KWS of which I am grateful. The course was co-funded by the DuPont Agricultural Enterprise, USA and a Sandwich arrangement between ICIPE and the JKUAT. I am sincerely grateful for the financial support. Finally, I sincerely thank my family for their patience and support for the entire period of my study v ACNONYMS AND ABBREVIATIONS BLAST Basic Local Alignment Search Tool cDNA Complimentary DNA Da Dalton DDD Dichlorodiphenyldichloroethane DDT Dichlorodiphenyltrichloroethane DNA Dioxyribonucleic acid DPTU Diphenylthiourea DPU Diphenylurea EPA Environmental Protection Agency ESI Electron Spray Ionisation FDA Food and Drug Administration GC Gas Chromatography HPLC High Performance Liquid Chromatoraphy ICIPE International Centre of Insect Physiology and Ecology JKUAT Jomo Kenyatta University of Agriculture and Technology KWS Kenya Wildlife Services LC Liquid Chromatography LC;0 Concentration that would kill 50% of test organism LD;o Dose that would kill 50% of test organism MALDI Matrix Assisted Laser Desorption Ionisation MS Mass Spectrometry AMNH American Museum of Natural History NMR Nuclear Magnetic Resonance Plit Parabuthm leiosoma insect toxin Plmt Parabuthus leiosoma mammalian toxin Plt Parabuthus leiosoma toxin PMTC N-Phenyl,o-Methyl-thiocarbonate TCEP Tris (2-carboxyethyl) phosphine TDE 1,1-Dichloro-2, 2-bis (p-chlorophenyl) ethane TOF Time of Flight vi TABLE OF CONTENTS Title Page Title ..................................... ........... ........................... .......... ............ ....... .................................. ................ .. 1 Declarations ........... ..... .......... .. ..... ............ .................................................................................... ........ ... .... ... ii Dedications ........ .. ............ .......... .. ... ............................................ ....................... ... .......... .............................. iii Acknowledgement ................ ................. ............................................................................... ..................... .iv Acronyms and abbreviations ............................................... .. ... ..................... ........... ..... ........ .. ............ ....... v Table of contents ........................ ... .............................................................................. ......................... ....... vi List of figures .............................. ........ .... ........................... .................. .... ... .. .. ..... ................. ............... ......... x List of tables ..... ... .......................... .......... ..................... ..................... ............................................................ x List of plates ................ ................... ................... ........... ...................... ........ ....... ......................... ... ............... xi List of schemes ............................................... ...................................... .. ........ ............................................ xii Appendices ... ............. .... .......... .. ............ ................... ... ... .. .......................... .. .... .. .............. .................. ... ...... xii Abstract .............. ... ............... ... ... .. ..................... ........... ... ............ ............ .. .............................................. .. xiii CHAPTER ONE: INTRODUCTION ......................................................... 1 1.1 Background ......................... ... ........ ....................... ....................................... .............. .. ... ... ................. .. 1 1.2 Sustainable economic and environmental development .. .. ...... ..... ........ ............................... ..... ..... 2 1.3 Chilo partel/11s (Swihoe) ..... ........................................................................ ............................................. 4 1.4 Control of insect pests ........... ...... ..... .. .. .......... ... ..................... ..... ,.. ..... .. ................. .... .. ..... .... .. ...... .. ... .4 1.4.1 Cultural practices .. ..... ........... .. ....... ......... ...... .. ... ........... ........................ .. .. ... ............ .... .. .... .. .. 4 1.4.2 Biological control ....... .. ... .... .. ...... ... ................... ......................... ........ ...................... ......... .. .... 5 1.4.2.1 Parasitoids ..... ....... ................... ......................... ................... ................ .................... 5 1.4.2.2 Predators .. .... .............................. ................................. .......... .................................. 6 1.4.2.3 The 'push' and 'pull' strategy ........................................................ .... ... ................ 6 1.4.2.4 Biotechnology ... ... ............. ... ............ ..... .......... .. .. .......................................... ... ...... 8 1.4.3 Insecticides ......... ........ ............... ............................................................................................... 9 1.5 Natural products for human welfare ....... ... ...... ...... ...... ............. ................. ...................... .............. 13 1.5.1 Arthropod defense secretions ............................................................................... ..... ..... ... 13 VII CHAPTER TWO: LITERATURE REVIEW .............................................. 15 2.1. The Scorpion ..................................................................................................... .. ... .. ....................
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