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Identification of Ixodes Ricinus Female Salivary Glands Factors Involved in Bartonella Henselae Transmission

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Identification of Ixodes Ricinus Female Salivary Glands Factors Involved in Bartonella Henselae Transmission UNIVERSITÉ PARIS-EST École Doctorale Agriculture, Biologie, Environnement, Santé T H È S E Pour obtenir le grade de DOCTEUR DE L’UNIVERSITÉ PARIS-EST Spécialité : Sciences du vivant Présentée et soutenue publiquement par Xiangye LIU Le 15 Novembre 2013 Identification of Ixodes ricinus female salivary glands factors involved in Bartonella henselae transmission Directrice de thèse : Dr. Sarah I. Bonnet USC INRA Bartonella-Tiques, UMR 956 BIPAR, Maisons-Alfort, France Jury Dr. Catherine Bourgouin, Chef de laboratoire, Institut Pasteur Rapporteur Dr. Karen D. McCoy, Chargée de recherches, CNRS Rapporteur Dr. Patrick Mavingui, Directeur de recherches, CNRS Examinateur Dr. Karine Huber, Chargée de recherches, INRA Examinateur ACKNOWLEDGEMENTS To everyone who helped me to complete my PhD studies, thank you. Here are the acknowledgements for all those people. Foremost, I express my deepest gratitude to all the members of the jury, Dr. Catherine Bourgouin, Dr. Karen D. McCoy, Dr. Patrick Mavingui, Dr. Karine Huber, thanks for their carefully reviewing of my thesis. I would like to thank my supervisor Dr. Sarah I. Bonnet for supporting me during the past four years. Sarah is someone who is very kind and cheerful, and it is a happiness to work with her. She has given me a lot of help for both living and studying in France. Thanks for having prepared essential stuff for daily use when I arrived at Paris; it was greatly helpful for a foreigner who only knew “Bonjour” as French vocabulary. And I also express my profound gratitude for her constant guidance, support, motivation and untiring help during my doctoral program. She has always been nice to me and made herself available to clarify my doubts despite her busy days. During my research, she gave me enough freedom to do many projects without objection. In a word, I could not have completed my doctoral thesis without her. I express my gratitude to Dr. Muriel Vayssier-Taussat, Director of “Vectotic” group. She gave me many insightful suggestions for the experiments, thesis writing, and presentation. I also thank her for completing the manuscript of my thesis. I am thankful to Dr. Sara Moutailler. It was a pleasure to share the office with her and she gave me many scientific advices for the experiments, especially quantitative PCR performing and results analysis process. She was kind to share her ticks with me. Thanks for her contribution to my manuscripts, too. Many thanks to Danielle Le Rhun who has always been ready to help me, when I was in trouble not only for the experiments but also for my life in Paris. And she gave many scientific advices to my experiments, and did a lot of work for my thesis. I also 1 thank her for correcting my French speaking and writing. Without her help, I could not have finished my PhD projects on time. I would like to thank Martine Cote. She engorged many ticks for me during my doctoral program, and sometimes had to work during weekends. I am grateful to Evelyne Le Naour. Thanks for her help in reagents preparation and for inviting me in her house for my holidays. It’s my pleasure to acknowledge all the colleagues in “USC-INRA Bartonella et Tiques” and “Mission Tiques”, especially Jean-Philippe Buffet, Dominique Huet, Françoise Féménia, Julien Chotte, Lorraine Michelet, Elodie Devillers, Thibaud Dugat, for their supporting and providing a good atmosphere in the laboratory and office. I’m very grateful to them for helping me to complete my PhD studies. I also thank our secretaries Matthieu Chaumien, Viviane Domarin, and Sophie Hourigat, who took care of all official works, including products orderings. I also express my gratitude to Dr. Richard Paul (Institute Pasteur) and Prof. José de la FUENTE (Oklahoma State University). They gave me many insightful suggestions for the experiments. I thank Director Pascal Boireau, Director Nadia Haddad, Prof. Jacques Guillot， Dr. Sandrine Lacour for their help on my thesis. They are always very kind to all the Chinese students in UMR BIPAR. I also would like to thank the Director of CRBM, Thomas Lilin, and all his colleagues: Benoît Lécuelle, Serge Kouame, Cathy Claramonte, and Ingrid Gruyer for taking care of animals used in my PhD studies. And there’re also many thanks to Alain Bernier, Océane Le Bidel, Sébastien Allix for animal helping. I thank all the present members of SEPPIC group: Elodie Carneaux, Nicolas Versillé, Anna Rosemond, and Jennifer Maye. The acknowledgements are for their help and friendship to me during the past four years. Two persons, who took care of non-scientific works, but are very important to me: thank to Dany Espuche and Madeleine Carle for washing all the materials and 2 autoclave. I thank the present people of doctoral school in Université Paris-Est: Director Cyril Kao (ABIES), Director Alain Berdeaux (SVS), and Secretary Corinne Fiers (ABIES), Secretary Candice Gottscheck (SVS). Thanks for their help in my inscription in Université Paris-Est. I thank my friends Dr. Hongkuan Deng, Dr. Xiaocui Zhou, Dr. Yong Yang, and Dr. Dongying Wang. We all studied in UMR BIPAR and discussed many issues on laboratory and experiment topics. Thanks for their propositions for my thesis. I also would like to thank my friends Yiming Xiao, Limin Meng, Qiongfei Xu, Ke Wang. We have been the members of a Chinese students unit in Paris named “Association de Boursiers CSC en Ile de France”. We organized many travels in France, that let me well understand the history and culture of this country and made my life meaningful and wonderful. There are too many friends I want to thank but not listed here. However, you know who you are. The acknowledgements for the present members of Jilin university: Prof. Mingyuan Liu, Dr. Xiuping Wu, Prof. Xuelin Wang, Prof. Lu Yu, Director Guangming Wang, thanks for their encouragement. I will forever be thankful to CSC (China Scholarship Council), thanks for the scholarship supporting throughout my 48 months PhD studies in Paris. I also thank the EDENext Steering Committee for fund providing and the “Tiques et Maladies à Tiques” working group (REID- Réseau Ecologie des Interactions Durables) for thesis suggestions. I would like to be grateful to all the members of the Office of Educational Affairs of the Embassy of P.R. China in France, especially Minister Counselor Ms. Xiaoyu Zhu, Officer Ms. Yaping Qiang, and Officer Ms. Jingmei Zhao; thanks for their help and encouragement. Last but not the least I would like to thank my family. My parents provided unconditional love and care to me. I would not have made it so far without them. 3 TABLE OF CONTENTS ACKNOWLEDGEMENTS .......................................................................................... 1 TABLE OF CONTENTS .............................................................................................. 4 I. INTRODUCTION ................................................................................................... 5 II. BACKGROUND.................................................................................................... 8 II.1. IXODES RICINUS ...................................................................................................... 8 II. 1.1. Taxonomy and morphology ................................................................................... 8 II. 1.2. Geographical distribution .................................................................................... 11 II. 1.3. Biological cycle ..................................................................................................... 12 II.1.4. Pathogens transmitted by I. ricinus ...................................................................... 14 II.2. BARTONELLA SPECIES AND TICK BORNE TRANSMISSION .................................................. 16 II.3. TICKS REARING AND INFECTION METHODS .................................................................. 19 II.4. TICK-BORNE PATHOGEN TRANSMISSION: MODALITIES AND MOLECULAR MECHANISMS ......... 32 II.4.1. General introduction ............................................................................................ 32 II.4.2. Tick molecules implicated in pathogen transmission ........................................... 33 II.5. TBD VACCINE STRATEGIES BASED ON TICK MOLECULES .................................................. 69 III. OBJECTIVES ..................................................................................................... 71 IV. EXPERIMENTAL STUDIES ................................................................................. 72 IV.1. EVALUATION OF MEMBRANE FEEDING FOR INFECTING I. RICINUS WITH BARTONELLA SPP. .... 72 IV.1.1. Introduction to article 1 ...................................................................................... 72 IV.1.2. Article 1 ............................................................................................................... 73 IV.1.3. Conclusion of article 1 ......................................................................................... 94 IV.2. ANALYSIS OF B. HENSELAE-INFECTED I. RICINUS SALIVARY GLAND TRANSCRIPTS ................. 95 IV.2.1. Introduction to article 2 ...................................................................................... 95 IV.2.2. Article 2 ............................................................................................................... 98 IV.2.3. Conclusion of article 2 ....................................................................................... 138 V. DISCUSSIONS AND CONCLUSIONS ..................................................................
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