EVALUATING ONCHOCERCIASIS USING PARASITOLOGICAL, ENTOMOLOGICAL AND SLIDE AGGLUTINATION TECHNIQUE IN KACHIA, KADUNA STATE, NIGERIA BY Hudu Okankhamame OSUE, B.Sc. (BENSU, 1987), M.Sc. (ABU, 1996) Ph. D/Scien/19948/07-08. A THESIS SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF A DOCTOR OF PHILOSOPHY IN MICROBIOLOGY DEPARTMENT OF MICROBIOLOGY, FACULTY OF SCIENCE, AHMADU BELLO UNIVERSITY, ZARIA, NIGERIA JULY, 2015 i DECLARATION I declare that the work in this Thesis entitled "Evaluating Onchocerciasis Using Parasitological, Entomological and Slide Agglutination Technique in Kachia, Kaduna State, Nigeria" has been carried out by me in the Department of Microbiology, Faculty of Science, Ahmadu Bello University, Zaria under the supervision of Prof. (Mrs.) H. I. Inabo, and Prof. S. E. Yakubu, of Microbiology Department and Prof. P. A. Audu, of the Department of Biological Sciences. The information derived from the literature has been duly acknowledged in the text and a list of references provided. No part of this dissertation was previously presented for another degree or diploma at this or any other Institution. ______________________________ __________________ _______________ Hudu Okankhamame OSUE Signature Date ii CERTIFICATION This Thesis entitled EVALUATING ONCHOCERCIASIS USING PARASITOLOGICAL, ENTOMOLOGICAL AND SLIDE AGGLUTINATION TECHNIQUE IN KACHIA, KADUNA STATE, NIGERIA by Hudu Okankhamame OSUE meets the regulations governing the award of the degree of Doctor of Philosophy in Microbiology of Ahmadu Bello University, and is approved for its contribution to knowledge and literary presentation. Prof. H. I. Inabo_____________ ___________________ ______________ Chairman, Supervisory Committee (Signature) Date Prof. S. E. Yakubu______________ ____________________ ______________ Member, Supervisory Committee (Signature) Date Prof. P. A. Audu______________ _____________________ ______________ Member, Supervisory Committee (Signature) Date Prof. S. A. Ado _________________ _____________________ ______________ Head of Department (Signature) Date Prof. K. Bala__________________ _____________________ ______________ Dean, School of Postgraduate Studies (Signature) Date iii ACKNOWLEDGEMENTS I am immensely grateful to members of my supervisory team: Prof. H. I. Inabo and Prof. S. E. Yakubu of the Department of Microbiology and Prof. P. A. Audu of Department of Biological Sciences for the articulate academic guide and morale boosting they provided to me. My thanks go to the Head of Department, Prof. S. A Ado, Prof. O. S. Olonitola, immediate past Postgraduate Coordinator, Prof. I. O. Abdullahi, the Postgraduate Coordinator, Dr. D. Machido and the other esteemed and respected Lecturers of the Department of Microbiology. My appreciation goes to Prof. E. C. Okolocha and Dr. Kabiru Junaidu of Department of Veterinary Public Health and Preventive Medicine. My affection and gratitude goes to my wife, Hajia Bilkisu Adamu Hudu and my children, Usman, Seudat, Khadijat, Mohammed, Buithat, Medinat, and Naimat for their support and understanding. I thank my brothers, Mr. Musa I. Oshionei, Zakari Osue, Alh Idris Bako Osue and my sisters, Fatimetu, Hajia Omomo Uthman and Rametu for their encouragement. My gratitude goes to the Director General/CEO of NITR, Prof. M. Mamman for sponsorship and funding of the research projects. Many thanks to the former Director of Administration and Finance, Alhaji (Dr.) A. A. Dangaladima, the first Tukura Gwandu, the entire Management and staff of NITR, my colleagues, Mallam Sule Enyisi and fellow postgraduate students for their cordial relationship. I whole-heartedly appreciate the assistance rendered by the Chairman and Director of Health, Kachia Local Government Area, the people and Village Heads of Bomjock, Gantan, Gidan Tama, Kurmin Gwaza and Unguwar Shaho for their warm reception, cooperation and voluntary participation without which this thesis would not have been possible. I wish to register my appreciation to Dr. Gloria Chechet for her immense role in the PCR assay. Above all, I thank Almighty Allah (SWT), the beneficent and the most merciful for given me the grace, the health and wherewithal to embark and complete this course. iv DEDICATION This dissertation is dedicated to the evergreen and blessed memory of my parents, Madam Rabiet and Mallam Saidu Osue, my maternal uncle and guardian, Lieutenant Abdul Braimah Lawrence, and brother, Prince Ahmed Momodu Ikhesi, and my immediate younger sister, Aminat Amedu (Nee Ayoshi). May the Almighty Allah (SWT) grant them Aljauna Firdausi (Ameen). To all the blind and visually impaired people the world over. v Table of Contents Page Title page------------------------------------------------------------------------ i Approval Page------------------------------------------------------------------- iii Acknowledgements------------------------------------------------------------- iv Dedication------------------------------------------------------------------------ v Table of contents---------------------------------------------------------------- vi List of Tables-------------------------------------------------------------------- xi List of Figures------------------------------------------------------------------- xii List of Plates--------------------------------------------------------------------- xiii List of Appendices-------------------------------------------------------------- xiv Abbreviations-------------------------------------------------------------------- xv Definitions------------------------------------------------------------------------ xviii Glossary--------------------------------------------------------------------------- xx Abstract--------------------------------------------------------------------------- xxii 1.0 INTRODUCTION----------------------------------------------------------- 1 1.1 Preamble----------------------------------------------------------------------- 1 1.2 Statement of the Research Problem-------------------------------------- 3 1.3 Justification of the Study--------------------------------------------------- 4 1.4 Theoretic framework-------------------------------------------------------- 5 1.5 Aims and objectives of the study------------------------------------------ 6 1.6 Research Questions----------------------------------------------------------- 7 2.0 LITERATURE REVIEW-------------------------------------------------- 8 2.1 Epidemiology of Onchocerciasis------------------------------------------ 12 2.2 Geospatial Data and Vector-Host Interaction------------------------- 13 vi 2.3 Disease Transmission------------------------------------------------------- 15 2.4 Immunopathology of Onchocerciasis------------------------------------ 16 2.5 Clinical Manifestations of Onchocerciasis------------------------------ 17 2.6 Serodiagnosis of Onchocerciasis------------------------------------------ 18 2.7 Current Control of Onchocerciasis-------------------------------------- 19 2.7.1 Community directed treatment with ivermectin (CDTI)---------------- 20 2.7.2 Possibility of development of drug resistance to ivermectin------------ 21 2.7.3 Macrofilaricidal action of ivermectin--------------------------------------- 22 2.7.4 Expansion of community directed treatment with ivermectin----------- 22 2.7.5 Impact of CDTI on onchocerciasis epidemiological status-------------- 25 2.7.6 Effect of ivemectin treatment on adult worm ----------------------------- 26 2.7.7 Criteria for cessation of ivermectin treatment----------------------------- 27 2.8 Research and Development for Macrofilaricides--------------------- 27 2.8.1 Macrofilaricidal activities of some antibiotics---------------------------- 29 2.8.2 Moxidectin clinical trials and commercialization------------------------ 30 2.9 Wolbachia Bacterial Endosymbiont of Onchocerca volvulus------- 31 2.10 Strategy for Onchocerciasis Drug Discovery Initiative-------------- 33 2.11 Diagnostic Challenges in Onchocerciasis Control-------------------- 35 2.11.1 Immunodiagnostic tests------------------------------------------------------ 36 2.11.2 Nucleic acid based test------------------------------------------------------- 37 2.11.3 Application of recombinant technology----------------------------------- 39 2.11.4 Enzymes as diagnostic biomarkers---------------------------------------- 40 2.11.5 Metabolomics diagnostic biomarkers for onchocerciasis---------------- 41 2.11.6 Importance of antibody based serodiagnosis of onchocerciasis--------- 42 2.12 Bioinformatics and Databases for Onchocerciasis-------------------- 43 vii 2.12.1 Role of expressed sequence tag in onchocerciasis------------------------ 45 2.12.2 Possible areas of application of expressed sequence tag----------------- 45 3.0 MATERIALS AND METHODS------------------------------------------ 47 3.1 The study Area--------------------------------------------------------------- 47 3.2 The Study Population------------------------------------------------------- 48 3.3 Bio-clinical Data-------------------------------------------------------------- 48 3.4 Socio-Economic Activities of the Study Area--------------------------- 49 3.5 Geospatial and Entomology Data----------------------------------------- 49 3.6 Experimental Design--------------------------------------------------------- 53 3.7 Field Epidemiological Surveys--------------------------------------------- 55 3.8 Case Definition of Onchocerciasis----------------------------------------- 55