Bioprospecting for Anti-Mosquito Phytochemicals Associated with Olfaction and Larvicidal Activities from Selected Kenyan Plants
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i BIOPROSPECTING FOR ANTI-MOSQUITO PHYTOCHEMICALS ASSOCIATED WITH OLFACTION AND LARVICIDAL ACTIVITIES FROM SELECTED KENYAN PLANTS . JOHN BWIRE OCHOLA (MSc. B.Sc.) I84/27752/2013 A THESIS SUBMITTED IN FULFILMENT OF THE REQUIREMENT FOR THE AWARD OF THE DEGREE OF DOCTOR OF PHILOSOPHY (CHEMISTRY) IN THE SCHOOL OF PURE AND APPLIED SCIENCE OF KENYATTA UNIVERSITY JANUARY 2020 ii DECLARATION I certify that this thesis is my original work and has not been presented for award of a degree in any other university. Besides where due acknowledgement has been made, the work is the result of my labour which has been accomplished from the commencement date of the permitted studies application. Signature ……………………………… Date ………………………………… John Bwire Ochola Reg I84/27752/2013 Department of Chemistry Declaration by supervisors We confirm that the work reported in this thesis was carried out by the student under our supervision and has been submitted for examination with our approval as supervisors Signature ……………………………………. Date …………………… Prof. Ahmed Hassanali Department of Chemistry, Kenyatta University, Nairobi, Kenya Signature ……………………………………. Date …………………… Dr. Wilber Lwande Applied Bioprospecting Programme International Centre of Insect Physiology and Ecology, Nairobi, Kenya. Signature ……………………………………. Date …………………… Dr. Barbara Frei Haller Institute of Pharmaceutical Sciences, ETH Zurich Switzerland iii DEDICATION “To my wife Jackline and sons, Nathan, Seth, Lawrence and Ronnie, you kept my spirit alive.” iv ACKNOWLEGDEMENTS I express my sincere gratitude to Dr Wilber Lwande of ICIPE, Prof. Ahmed. Hassanali of the Department of Chemistry, Kenyatta University and Dr. Barbara Frei Haller of the Institute of Pharmaceutical Sciences, ETH Zurich, Switzerland for their support and mentorship throughout the research period and in the writing of this thesis. I would also like to thank Dr Margret Ng’ang’a for her guidance and support. My sincere thanks go to Dr Patrick Guerin and Thomas Krober for hosting me at Neuchâtel University, Switzerland and for providing practical knowledge on GC- EAD technique. Dr. Pie Muller for hosting me at the Swiss Tropical and Public Health Institute, Switzerland, and sharing enthusiastically his experiences in Vector Control Research, mainly on repellent testing. My most sincere appreciation goes to Dr Barbara Frei Haller for her dedication to fund my tuition fee without whose support I would not have come this far. Special thanks also go to UBS Optimus Foundation for funding part of my research work within the African Repellents Project. ICIPE is highly appreciated for provision of the research environment. I am equally indebted to my colleagues; Mr. Wanyama for carrying out GC-MS analyses of my samples, Rose Marubu, Nixon Onyimbo, Fred Nduguli and Lamberts Moreka at the Bioprospecting Programme at ICIPE. I owe special gratitude to my family particularly my wife, Jackline Majuma Bwire for her love, support and patience mixed with a dose of “go for it!” Lots of thanks also go to my dear parents (posthumous), siblings and friends. Finally, I give all glory and honor to God Almighty who continues to guide, sustain and carry me through life’s challenges. v TABLE OF CONTENT DECLARATION ................................................................................................................. ii DEDICATION .................................................................................................................... iii ACKNOWLEGDEMENTS ............................................................................................... iv TABLE OF CONTENT ...................................................................................................... v LIST OF FIGURES ............................................................................................................ xi LIST OF PLATES ............................................................................................................ xiii LIST OF SCHEMES ........................................................................................................ xiv LIST OF APPENDICES ................................................................................................... xv CHAPTER ONE .................................................................................................................. 1 INTRODUCTION ............................................................................................................... 1 1.1 Background information ............................................................................................ 1 1.2 Statement of the Problem ........................................................................................... 5 1.3 Justification of the Study ........................................................................................... 5 1.4 Significance of the study........................................................................................... 6 1.5 Hypotheses ................................................................................................................. 6 1.6 General objective ....................................................................................................... 7 1.6.1 Specific objectives ..................................................................................................... 7 1.7 Scope and limitation ................................................................................................. 8 CHAPTER TWO ................................................................................................................. 9 LITERATURE REVIEW ................................................................................................... 9 2.1 Mosquitoes as vectors of diseases ............................................................................. 9 2.2 Biology and life cycle of a mosquito ....................................................................... 11 2.3 Economic importance of mosquitoes ....................................................................... 13 2.4 Host-seeking behaviour of mosquito ....................................................................... 13 2.5 Mosquito vector borne disease burden .................................................................... 16 2.5.1 Malaria ..................................................................................................................... 17 2.5.2 Dengue ..................................................................................................................... 18 2.5.3 Chikungunya ............................................................................................................ 18 vi 2.5.4 Yellow fever ............................................................................................................ 19 2.5.5 Zika virus disease..................................................................................................... 19 2.5.6 West Nile virus ........................................................................................................ 20 2.6 Distribution of mosquito borne diseases .................................................................. 20 2.7 Control of mosquito vectors .................................................................................... 22 2.7.1 Chemical control of mosquitoes .............................................................................. 22 2.7.2 Environmental control ............................................................................................. 25 2.7.3 Biological/Genetic Control ...................................................................................... 25 2.7.4 Vector Resistance .................................................................................................... 26 2.8 Use of Natural products in Mosquito Control ........................................................ 27 2.8.1 Mosquito Repellents ................................................................................................ 28 2.8.2 Larvicides................................................................................................................. 31 2.9 Plants used in the study ............................................................................................ 32 2.9.1 The lamiaceae family ............................................................................................... 32 2.9.1.1 The genus Satureja .................................................................................................. 32 2.9.1.1.1 Biological studies and clinical properties of Satureja Species ........................ 33 2.9.1.1.2 Previous phytochemical studies of Satureja species ....................................... 33 2.9.1.2 The genus Plectranthus ........................................................................................... 34 2.9.1.3 The genus Ocimum .................................................................................................. 35 2.9.1.3.1 Ocimum kilimandscharicum ............................................................................ 35 2.9.1.3.2 Ethnobotany of O. kilimandscharicum ............................................................ 35 2.9.2 The family Asteraceae ............................................................................................. 38 2.9.2.1 The genus Conyza .................................................................................................... 38 2.9. Terpenoid biosynthesis ...........................................................................................