University of Veterinary Medicine Hannover Crimean-Congo hemorrhagic fever orthonairovirus (CCHFV): Surveillance studies among different livestock in sub-Saharan Africa and the molecular characterization of Hyalomma ticks serving as main reservoir and vector Inaugural-Dissertation to obtain the academic degree Doctor medicinae veterinariae (Dr. med. vet.) submitted by Ansgar Schulz Wismar Hannover 2020 Academic supervision: Prof. Dr. Martin H. Groschup Friedrich-Loeffler-Institut Federal Research Institute for Animal Health Institute of Novel and Emerging Infectious Diseases Greifswald-Insel Riems 1. Referee: Prof. Dr. Martin H. Groschup Friedrich-Loeffler-Institut Federal Research Institute for Animal Health Institute of Novel and Emerging Infectious Diseases Greifswald - Insel Riems 2. Referee: Prof. Dr. Christina Strube Institute of Parasitology University of Veterinary Medicine Hannover Hannover Day of the oral examination: 16th June 2020 - Sponsorship: This work was supported by the German Federal Foreign Office in the framework of the German Partnership Program for Excellence in Biological and Health Security. TABLE OF CONTENTS 1 INTRODUCTION ...........................................................................01 2 LITERATURE REVIEW ................................................................04 2.1 History and naming of CCHFV ..........................................................................04 2.2 Classification ......................................................................................................04 2.3 Morphology and genome characterization .......................................................05 2.3.1 S segment ……………………………………………………………………………….06 2.3.2 M segment ............................................................................................................06 2.3.3 L segment .............................................................................................................06 2.4 Virus replication ..................................................................................................07 2.5 Transmission ......................................................................................................08 2.6.1 Hylomma ticks and their special role in CCHFV transmission cycle ……………….08 2.6.2 Tick transmission routes …....................................................................................10 2.6.3 Exposure risks in humans/ direct transmission ......................................................11 2.6 Epidemiology ......................................................................................................12 2.6.1 Europe ……………………………………………………………………………….….12 2.6.2 Asia .......................................................................................................................14 2.6.3 Africa ....................................................................................................................14 2.7 Clinical signs in humans ….................................................................................16 2.8 Pathology ............................................................................................................17 2.9 Diagnostic and detection of CCHFV ..................................................................17 2.10 Prevention and treatment ...................................................................................19 3 MATERIAL AND METHODS ........................................................21 3.1 Samples…………………………. ..........................................................................21 3.2 Serology……. ......................................................................................................21 3.3 DNA/RNA extraction………………………… .......................................................22 3.4 Morphological tick species identification………………….................................22 3.5 Molecular tick species identification……….......................................................22 3.6 Virus detection and characterization……..........................................................23 3.7 Statistical analyzes…………………………..........................................................23 3.8 Literature research…………………………..........................................................24 4 MANUSCRIPT I: Crimean-Congo hemorrhagic fever orthonairovirus antibodies in Mauritanian livestock (cattle, goats, sheep and camels) is stratified by the animal´s age..……......................................................................25 4.1 Abstract ...............................................................................................................25 4.2 Introduction ........................................................................................................26 4.3 Materials and methods ......................................................................................27 4.4 Results ….............................................................................................................30 4.5 Discussion ..........................................................................................................31 4.6 Conclusion ..........................................................................................................34 4.7 Tables ..................................................................................................................34 4.6 Figures ................................................................................................................37 4.8 Acknowledgments ..............................................................................................37 5 MANUSCRIPT II: Molecular discrimination of Hyalomma tick species serving as reservoirs and vectors for Crimean-Congo hemorrhagic fever virus in sub-Saharan Africa ...........................................................................38 6 MANUSCRIPT III: Detection of Crimean- Congo hemorrhagic fever orthonairovirus (CCHFV) in Hyalomma ticks collected from Mauritanian livestock- more than just a virus indicator?..........................................................40 6.1 Abstract ...............................................................................................................40 6.2 Introduction ........................................................................................................41 6.3 Materials and methods ......................................................................................42 6.4 Results ................................................................................................................44 6.5 Discussion ..........................................................................................................47 6.6 Tables ..................................................................................................................50 6.7 Figures ................................................................................................................53 6.8 Acknowledgments ..............................................................................................54 7 GENERAL DISCUSSION .............................................................55 8 SUMMARY ...................................................................................61 9 ZUSAMMENFASSUNG ................................................................63 10 BIBLIOGRAPHY ..........................................................................66 11 AUTHORS´ CONTRIBUTION ......................................................80 12 ACKNOWLEDGMENTS ...............................................................82 Manuscripts extracted from the doctorate project: 1.) A. Schulz, Y. Barry, F. Stoek, A. Ba, M.A. Sas, J. Schulz, P. Kirkland, M. Eiden, M.H. Groschup Crimean-Congo hemorrhagic fever orthonairovirus antibody prevalence in Mauritanian livestock (cattle, goats, sheep and camels) are stratified by the animals age; to be submitted 2.) A. Schulz, A. Karger, B. Bettin, A. Eisenbarth, M. A. Sas, C. Silaghi, M. H. Groschup; Molecular discrimination of Hyalomma tick species serving as reservoirs and vectors for Crimean-Congo hemorrhagic fever virus in sub-Saharan Africa, Ticks and Tick-borne Diseases (2020) 101382 (https://doi.org/10.1016/j.ttbdis.2020.101382) 3.) A. Schulz, Y. Barry, F. Stoek, M. J. Pickin, A. Eisenbarth, M. Eiden, M. H. Groschup; Detection of Crimean- Congo hemorrhagic fever orthonairovirus (CCHFV) in Hyalomma ticks collected from Mauritanian livestock- more than just a virus indicator?; to be submitted Manuscripts excluded from the doctorate project: K. K. Kasi, F. v. Arnim, A. Schulz, A. Rehman, A. Chudhary, M. Oneeb, M. A. Sas, T. Jamil, P. Maksimov, C. Sauter‐Louis, F. J. Conraths, M. H. Groschup Crimean‐Congo haemorrhagic fever virus in ticks collected from livestock in Balochistan, Pakistan; Transboundary and Emerging Diseases (2020), (https://doi.org/10.1111/tbed.13488) K. K. Kasi, M. A. Sas, C. Sauter‐Louis, F. v. Arnim, J. M. Gethmann, A. Schulz, K. Wernike, M. H. Groschup, F. J. Conraths Epidemiological investigations of Crimean-Congo haemorrhagic fever virus infection in sheep and goats in Balochistan, Pakistan; Ticks and Tick-borne Diseases (2020) (https://doi.org/10.1016/j.ttbdis.2019.101324) R. Poueme, F. Stoek, N. Nloga, J. Awah-Ndukum, M. Rissmann, A. Schulz, A. Wade, J. Kouamo, M. Moctar, A. Eisenbarth, L. God-yang, S. Dickmu Seroprevalence and Associated Risk Factors of Rift Valley Fever in Domestic Small Ruminants in the North Region of Cameroon; Veterinary Medicine International (2019), (https://doi.org/10.1155/2019/8149897)