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Molecular Identification and Prevalence of Tick-Borne Pathogens in Zebu and Taurine Cattle in North Cameroon

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Molecular Identification and Prevalence of Tick-Borne Pathogens in Zebu and Taurine Cattle in North Cameroon Tick-borne pathogens in African cattle – novel molecular tools for diagnostics in epizootiology and the genetics of resistance Dissertation Der Mathematisch-Naturwissenschaftlichen Fakultät der Eberhard Karls Universität Tübingen zur Erlangung des Grades eines Doktors der Naturwissenschaften (Dr. rer. nat.) vorgelegt von Babette Josiane Guimbang Abanda aus Douala, Kamerun Tübingen 2020 Gedruckt mit Genehmigung der Mathematisch- Naturwissenschaftlichen Fakultät der Eberhard Karls Universität Tübingen. Tag der mündlichen Qualifikation: 17.03.2020 Dekan: Prof. Dr. Wolfgang Rosenstiel 1. Berichterstatter: PD Dr. Alfons Renz 2. Berichterstatter: Prof. Dr. Nico Michiels 2 To my beloved parents, Abanda Ossee & Beck a Zock A. Michelle And my siblings Bilong Abanda Zock Abanda Betchem Abanda B. Abanda Ossee R.J. Abanda Beck E.G. You are the hand holding me standing when the ground under my feet is shaking Thank you ! 3 Acknowledgements Finalizing this doctoral thesis has been a truly life-changing experience for me. Many thanks to my coach, Dr. Albert Eisenbarth, for his great support and all the training hours. To my supervisor PD Dr. Alfons Renz for his unconditional support and for allowing me to finalize my PhD at the University of Tübingen. I am very grateful to my second supervisor Prof. Dr. Oliver Betz for supporting me during my doctoral degree and having always been there for me in times of need. To Prof. Dr. Katharina Foerster, who provided the laboratory capacity and convenient conditions allowing me to develop scientific aptitudes. Thank you for your encouragement, support and precious advice. My Cameroonian friends, Anaba Banimb, Feupi B., Ampouong E. Thank you for answering my calls, and for encouraging me. I am grateful. Those who supported me in different ways, Dr. Adrian Streit, Prof. Dr. Jörn Bennewitz, Dr. Nils Anthes, Dr. Sina Beier, Dr. Volker Heiser, Prof. Dr. Nico Michiels. Those who always had a kind word, cheering me up late Dr. Almeck K. Aboubakar, Prof. Dr. Elias Nukenine, Prof. Dr. Tchuenguem F., Prof. Dr. Albert Ngakou, Prof. Dr. Mamoudou Abdoulmoumini, Dr. Wolfgang Hoffmann, Prof. Dr. Dieudonne Ndjonka, late Dr. Ombiono Messine. All the strong women who made it possible in their own way: Beck A Zock A., Prof. Dr. Elizabeth Ngo Bum. M. Boubakari for his support during the sampling. All the team of the station Programme Onchocercoses in Ngaoundéré, Cameroon for their support: David Ekale, Jeremie Yembo, Kalip Mbaiyanbe, late George Tamenai, late Luther Boudjepto. The staff of the Institute of Agricultural Research for Development (IRAD Wakwa, Cameroon): Dr. Kingsley T. Manchang, Ms. Maimounatou, and the students from the School of Veterinary Medicine and Sciences, at the University of Ngaoundéré in Cameroon. My colleagues and German friends Daniela Renz, Stephanie Maier, Dr. Fernanda Ruiz Fadel, Alexander Schinko, Mrs. Christina Nitsche, for their support and friendship. To all the funding bodies who contributed to the finalization of my doctorate thesis, I am deeply grateful: Boehringer Ingelheim Foundation, DAAD, Otto Bayer Foundation, Baden Württemberg Stiftung, Ministry of Science, Research and Arts Baden Württemberg, University of Tübingen (T@T) and Deutsche Forschungsgemeinschaft (Re 1536/4-1 & 1536/5-1). 4 Table of content Acknowledgements .................................................................................................... 4 Abbreviations .............................................................................................................. 7 Zusammenfassung ................................................................................................... 10 Résumé .................................................................................................................... 11 Disclosure of personal contribution ........................................................................... 12 Own contribution to the published paper 1: ........................................................... 12 Own contribution to the published paper 2: ........................................................... 12 Own contribution to the manuscript under finalization: .......................................... 13 List of publications .................................................................................................... 13 Included in the thesis: ............................................................................................ 13 Not included in the thesis: ..................................................................................... 14 General Introduction ................................................................................................. 15 A. The biology of ticks ......................................................................................... 15 1. Taxonomy, morphology and life cycle ............................................................. 15 2. Role as infectious disease vector ................................................................... 17 3. Pathogen transmission among tick host ......................................................... 18 3.1. Stadial transmission .................................................................................... 18 3.2. Transgenerational transmission .................................................................. 19 3.3. Co-feeding transmission ............................................................................. 20 3.4. Transmission by coxal fluid (Argasidae) ...................................................... 20 4. Interaction interfaces ...................................................................................... 21 4.1. Tick – pathogen interface ............................................................................ 21 4.2. Host vertebrate – tick interface.................................................................... 24 4.3. Host vertebrate – pathogen interface .......................................................... 24 5. Selected tick-borne pathogens and their significance in the cattle host .......... 25 5.1. Tick-borne viruses ....................................................................................... 26 5.2. Tick-borne bacteria ..................................................................................... 26 5.3. Tick-borne protozoans ................................................................................ 30 5.4. Generic life cycle ......................................................................................... 33 6. Epizootiology of tick-borne pathogens in Cameroon ....................................... 35 6.1. Tick-borne pathogens in the cattle population in Cameroon ....................... 36 6.2. Diagnostic tools for identification of pathogens transmitted by ticks............ 36 6.3. Enzootic concept of the interfaces cattle – tick – pathogen ......................... 37 5 6.4. Pathogen prevalence and vector distribution .............................................. 38 6.5. Control of tick-borne pathogens .................................................................. 40 6.6. Host heterogeneity and associated factors in host phenotype .................... 40 6.7. Cameroon’s genetic diversity in cattle as a source of host resistance ........ 41 B. Objectives, methodologies, structure of doctoral research ............................. 44 C. Results and discussion ................................................................................... 45 Chapter 1. Epizootiology of tick-borne pathogens and host associated factors ........ 45 Related publication ............................................................................................ 45 Extended summary ............................................................................................ 45 Heterogeneous reactivity and host response ..................................................... 46 Host response and environmental changes ....................................................... 48 Partial conclusion, perspectives and limitations ................................................. 50 Chapter 2. Development of an universally applicable microarray for the identification of co-infected tick-borne pathogens .............................................................................. 52 Related publication ............................................................................................ 52 Extended summary ............................................................................................ 52 LCD-Array validation: Sensitivity and specificity ................................................ 53 Probe design ...................................................................................................... 55 Limit of detection and copy number ................................................................... 55 Adjusted PCR protocol and biotinylated primers ................................................ 55 Partial conclusion, perspectives and limitations ................................................. 56 Chapter 3. Molecular genetic variants associated with host resistance against tick- borne diseases ......................................................................................................... 57 Related publication ............................................................................................ 57 Extended summary ............................................................................................ 57 Variance component estimates .........................................................................
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