Aus dem Institut für Mikrobiologie und Tierseuchen des Fachbereichs Veterinärmedizin der Freien Universität Berlin Fly-based wildlife disease surveillance in remote tropical habitats – two exemplary targeted microorganisms with zoonotic potential: Bacillus cereus biovar anthracis and adenoviruses Inaugural-Dissertation zur Erlangung des Grades eines Doktors der Veterinärmedizin an der Freien Universität Berlin vorgelegt von Constanze Hoffmann Tierärztin aus Chur, Schweiz Berlin 2018 Journal-Nr.: 4043 Gedruckt mit Genehmigung des Fachbereichs Veterinärmedizin der Freien Universität Berlin Dekan: Univ.-Prof. Dr. Jürgen Zentek Erster Gutachter: Prof. Dr. Lothar H. Wieler Zweiter Gutachter: PD Dr. Fabian Leendertz Dritter Gutachter: Prof. Dr. Alex Greenwood Deskriptoren (nach CAB-Thesaurus): diptera, diseases control, epidemiology, bacillus anthracis, adenoviridae, polymerase chain reaction, africa Tag der Promotion: 25.06.2018 Contents List of Figures .................................................................................................................... III List of Tables ...................................................................................................................... IV List of Abbreviations ........................................................................................................... V 1. Introduction ................................................................................................................... 1 2. Background ................................................................................................................... 3 2.1 EMERGING INFECTIOUS DISEASES ............................................................................................. 3 2.2 WILDLIFE DISEASE SURVEILLANCE ............................................................................................ 6 2.2.1 Wildlife disease surveillance – from the beginnings to today ..................................... 6 2.2.2 Forms of wildlife disease surveillance ............................................................................ 7 General surveillance ..................................................................................................... 7 Targeted surveillance ................................................................................................... 7 2.2.3 Study design of targeted wildlife disease surveillance................................................... 8 2.2.4 Sample collection ............................................................................................................... 8 Invasive sampling ......................................................................................................... 9 Non-invasive sampling .............................................................................................. 10 Logistics ....................................................................................................................... 10 2.2.5 Data analysis and interpretation .................................................................................... 11 2.3 FLIES IN WILDLIFE DISEASE SURVEILLANCE ........................................................................... 14 2.3.1 Fly ecology ........................................................................................................................ 14 2.3.2 Fly species identification ................................................................................................. 15 2.3.3 Using fly-derived DNA for mammalian biodiversity assessment ............................ 15 2.3.4 Pathogen detection in flies ............................................................................................. 16 2.4 TAÏ NATIONAL PARK ................................................................................................................. 17 2.5 ANTHRAX ..................................................................................................................................... 19 2.5.1 Bacillus anthracis ................................................................................................................. 19 General characteristics ............................................................................................... 19 Epidemiology .............................................................................................................. 20 Pathogenesis ................................................................................................................ 21 Clinical manifestation ................................................................................................ 22 Bacillus anthracis in African wildlife ....................................................................... 24 Genotyping and evolution ........................................................................................ 25 2.5.2 Bacillus cereus biovar anthracis ........................................................................................... 26 2.6 ADENOVIRUSES........................................................................................................................... 29 2.6.1 General characteristics .................................................................................................... 29 2.6.2 Epidemiology and clinical manifestation ..................................................................... 29 2.6.3 Cross-species transmission and zoonotic potential .................................................... 30 3. Publications ................................................................................................................. 32 3.1 PUBLICATION I ............................................................................................................................ 32 3.2 PUBLICATION II ........................................................................................................................ 101 I CONTENTS 4. Discussion................................................................................................................... 111 4.1 BCBVA IN TNP: AN IN-DEPTH ANALYSIS ON SMALL SCALE ................................................ 112 4.1.1 Bcbva across space .......................................................................................................... 115 4.1.2 Bcbva across time ............................................................................................................ 116 4.2 BCBVA IN SUB-SAHARAN AFRICA: A FIRST LARGE-SCALE SCREENING .............................. 117 4.2.1 Sites with unknown Bcbva occurrence ........................................................................ 117 4.2.2 Sites with known Bcbva occurrence ............................................................................. 118 4.3 ADENOVIRUSES IN TNP: A FIRST SMALL-SCALE SCREENING ............................................. 121 4.4 FLY MEAL ANALYSIS ................................................................................................................. 123 4.4.1 Fly meal analysis and Bcbva in TNP ............................................................................ 123 4.4.2 Fly meal analysis and AdV in TNP ............................................................................. 125 4.5 FEASIBILITY OF USING FLIES FOR WILDLIFE DISEASE SURVEILLANCE .............................. 126 5. Conclusion and Outlook ............................................................................................. 128 6. Summary ..................................................................................................................... 129 7. Zusammenfassung ..................................................................................................... 130 8. References ................................................................................................................... 132 9. List of publications ..................................................................................................... 152 10. Acknowledgements..................................................................................................... 153 11. Selbstständigkeitserklärung ....................................................................................... 155 II List of Figures Figure 1: Geographic location of Taï National Park ............................................................................. 17 Figure 2: Sites in sub-Saharan Africa where Bcbva has been described ............................................. 27 Figure 3: Fly sampling on small scale in Taï National Park for publication I. ................................ 113 Figure 4: Fly sampling on large scale in sub-Saharan Africa for publication I. ............................... 117 Figure 5: Geographic location of Grebo National Forest (GNF) .................................................... 118 All figures were made by Constanze Hoffmann and not have been published elsewhere. III List of Tables Table 1: Forms of wildlife disease surveillance. .................................................................................... 13 Table 2: Samples and results for fly-based surveillance of Bcbva and AdV on small scale in Taï National Park (TNP), Côte d’Ivoire. .................................................................................................... 114 Table 3: Samples and results for fly-based disease surveillance of Bcbva on large scale in sub- Saharan Africa..........................................................................................................................................