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Prevalence of Bacterial Zoonoses in Selected Trophy Hunted

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Prevalence of Bacterial Zoonoses in Selected Trophy Hunted PREVALENCE OF BACTERIAL ZOONOSES IN SELECTED TROPHY HUNTED SPECIES, AND THE POTENTIAL OF HUMAN HEALTH RISK IN BWABWATA NATIONAL PARK, NAMIBIA A THESIS SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE IN BIODIVERSITY MANAGEMENT OF THE UNIVERSITY OF NAMIBIA BY AMEYA MATHEUS-AUWA (201030292) April 2019 Main Supervisor: Dr. Jean Damascène UZABAKIRIHO (Department of Biological Sciences, University of Namibia) Co-supervisor: Dr. Seth J. EISEB (Department of Biological Sciences, University of Namibia) ABSTRACT Zoonotic diseases are infections acquired from vertebrate animals (wild or domesticated) animals to humans through direct or indirect contact with live animals, their derivatives or contaminated surroundings. The aim of this study is to determine the prevalence of potential bacterial zoonoses in selected trophy hunted species Loxodonta Africana (African Elephant), Syncerus caffer (African Buffalo), Tragelaphus strepsiceros (Kudu), Hippopotamus amphibious (Hippopotamus), Hippotragus niger (Sable antelope), and Hippotragus equinus (Roan antelope), and the potential human health risk in Bwabwata National Park, North East Namibia. The Park covers an area size of 6 274 km2. It is divided in three Core Areas designated for wildlife conservation and controlled tourism namely: Kwando, Buffalo and Mahango core area and a large Multiple Use Area zoned for community-based tourism, trophy hunting, human settlement and development by the resident community. Forty-four tissue samples (kidney, heart, liver, spleen, lungs, and lymph nodes) were drawn from freshly shot carcasses of kudu (3), buffalo (2), hippo (1), roan (1), sable (1) and elephant (1). Blood Agar (BA), MacConkey Agar (McC) and Eosin Methylene Blue-agar (EMB) were used as culture media. A total of 16 isolates were obtained and identified using the Biochemical methods and the amplification of the 16S rRNA gene. All bacteria isolated are potential human pathogens. The prevalence 50% (8 of 16) of potential bacterial zoonoses was highest in Kudu followed by Sable 37.5% (6 of 16) and Hippo 31.25 % (5 of 16). While it was low in Buffalo 25% (4 of 16), Roan 18.75% (3 of 16) and Elephant 12.5% (2 of 16). Kidney tissues had the highest prevalence 68.8% (11 of 16), followed by liver 32.5% (10 of 16), heart 37.5% (6 of 16), lung 31.25% (5 of 16), spleen 31.25% (5 of 16) and lymph nodes 18.75% (3 of 16). Klebsiella pneumonia dominated the tissue organs 18.9% (recovered 8 times from 44 tissues), Micrococcus caseolyticus dominated the animals 66.7% (4 times from 6 animals). Phylogenetic relationship formed four clusters with 58-100% bootstrap values. This study demonstrated the prevalence of potential bacterial zoonoses in trophy hunted animals in BNP which might pose potential human health risk if transmitted. Trophy hunters and the BNP community should therefore be informed about the risks associated with trophy hunted animals and their derivatives. Keywords: Bacteria, Zoonoses, Trophy hunted animals, Bwabwata National Park i TABLE OF CONTENTS ABSTRACT ...................................................................................................................... i TABLE OF CONTENTS ................................................................................................ii LIST OF TABLES .......................................................................................................... v LIST OF FIGURES ....................................................................................................... vi LIST OF ABBREVIATIONS ......................................................................................vii ACKNOWLEDGEMENTS ........................................................................................... ix DEDICATION ................................................................................................................ xi DECLARATIONS ........................................................................................................xii CHAPTER 1: INTRODUCTION .................................................................................. 1 1.1 Orientation of the study ............................................................................................ 1 1.2 Statement of the problem ......................................................................................... 5 1.3 Main objectives of the study .................................................................................... 6 1.3.1 Specific objectives ............................................................................................. 6 1.4 Significance of the study .......................................................................................... 7 1.5 Limitations of the study ............................................................................................ 7 1.6 Delimitations of the study ........................................................................................ 7 CHAPTER 2: LITERATURE REVIEW ...................................................................... 8 2.1 Historical context of wildlife zoonotic diseases ....................................................... 8 2.2 The economic importance of the wildlife trophy hunting industry in Namibia ....... 9 ii 2.3 The wildlife - human-livestock continuum and its consequences .......................... 12 2.4 The physical continuum and the transmission of zoonotic bacterial infections ..... 14 2.4.1 Mechanisms of disease transmission ............................................................... 14 2.4.2 Infections of direct contact and their prevalence ............................................. 15 2.4.3 Infections of indirect contact and their prevalence .......................................... 18 2.4.4 Risk factors influencing potential exposure of bacterial zoonoses .................. 20 2.5 Major bacterial zoonotic diseases of health concern in Southern Africa ............... 22 2.5.1 Brucellosis ....................................................................................................... 24 2.5.2 Bovine tuberculosis or Tuberculosis (TB) ....................................................... 27 2.5.3 Anthrax ............................................................................................................ 31 CHAPTER 3: MATERIALS AND METHOD ........................................................... 34 3.1 Description of the study area and study population ............................................... 34 3.2 Research design ...................................................................................................... 36 3.3 Study species, sample collection, preparation and processing ............................... 36 3.4 Bacteria culture and isolation ................................................................................. 37 3.5 Morphology identification ...................................................................................... 38 3.6 Biochemical characterization ................................................................................. 38 3.7 DNA extraction and PCR amplification ................................................................. 41 3.8 Determination of the prevalence of potential zoonotic bacterial species ............... 42 3.9 Phylogenetic analysis ............................................................................................. 42 iii CHAPTER 4: RESULTS .............................................................................................. 44 4.1 Isolation and identification of isolated bacteria...................................................... 44 4.2 Prevalence of isolated bacteria from the six trophy hunted species (elephant, buffalo, kudu, sable, roan and hippo) in BNP .............................................................. 49 4.3 Molecular phylogenetic analysis of 16S rRNA gene sequences ............................ 50 CHAPTER 5: DISCUSSIONS ..................................................................................... 54 5.1 Identification of isolated bacteria, their prevalence and potential zoonoses .......... 54 5.2 Phylogenetic relationship of potential bacterial zoonoses isolated from trophy hunted animal from BNP .............................................................................................. 75 CHAPTER 6: CONCLUSIONS................................................................................... 78 CHAPTER 7: RECOMMENDATIONS ..................................................................... 79 REFERENCES .............................................................................................................. 80 APPENDICES ............................................................................................................. 118 Appendix 1: Biosafety plan ........................................................................................ 118 Appendix 2: Equipment management form ............................................................... 122 Appendix 3: Ethical clearance certificate for the study.............................................. 124 Appendix 4: Research or collecting permit 1 ............................................................. 125 Appendix 4: Research or collecting permit 2 ............................................................. 126 iv LIST OF TABLES Table 1: The number of the major wildlife species hunted by trophy hunters in Namibia in 2008 and 2009 (van Schalkwyk et al., 2010). .............................................................
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