Detection of Leptospira and Seasonal Prevalence of Fleas
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DETECTION OF LEPTOSPIRA AND SEASONAL PREVALENCE OF FLEAS COLLECTED FROM SMALL MAMMALS IN MUKWE CONSTITUENCY, KAVANGO EAST REGION OF NAMIBIA A THESIS SUBMITTED IN FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE (BIOLOGY) OF THE UNIVERSITY OF NAMIBIA BY SAIMA KAPIA 200948997 APRIL 2018 MAIN SURPERVISOR: DR. S. J. EISEB (UNIVERSITY OF NAMIBIA) CO-SUPERVISOR: PROF. L. S. MULUNGU (SOKOINE UNIVERSITY OF AGRICULTURE, TANZANIA) CO-SUPERVISOR: DR. P. TORTOSA (UNIVERSITÉ DE LA RÉUNION, FRANCE) Abstract Biotic and abiotic factors can alter the abundance and community composition of mammals and in turn that of associated parasites. The aim of the study was to measure the impact of climatic variables (temperature and relative humidity) on mammal populations and on the population dynamics of associated micro- and ecto-parasites. For this, the current study described prevalence and diversity of pathogenic Leptospira, the etiological agent of leptospirosis, and fleas, arthropod vectors of medical importance in mammals sampled over a period of 11 months in the Kavango East region of Namibia. In total, 121 small mammal hosts were examined for Leptospira using a molecular method (Multilocus sequence typing method) and yielded an overall prevalence rate of 9.9%. However, only a single Leptospira species could be identified as Leptospira kirschneri, on one host species (Saccostomus campestris). Throughout the study period, almost all mammal species harboured the same flea species, which included Cryptonella numae (1.8%), Pulex irritans (43.0%), Pariodontis riggenbachi riggenbachi (12.9%), Synosternus caffer (3.9%) and Xenopsylla species (38.3%). Using the Kruskal-Wallis test, the monthly population fluctuation of fleas varied significantly on M. natalensis (χ2=29.440, df=10, P=0.001), S. pratensis (χ2 =30.521, df=10, P=0.001) and S. campestris (χ2=32.681, df=10, P=0.0001). However, no significant difference in the number of fleas per month was found for G. leucogaster (χ2=10.831, df=10, P=0.371). The Pearson correlation test revealed that fleas abundance on small mammal hosts was weakly positively correlated with temperature (r=0.3, df=9, N=11, P=0.310) and strongly positively correlated with relative humidity (r=0.6, df=9, N=11, P=0.109), this same comparison for fleas showed a weak positive correlation (r=0.4, df=9, N=11, P=0.224). Lastly, there was a very strong positive correlation (r=0.8, df=9, N=11, P=0.0058) between average abundance of small mammals and average abundance of fleas. Therefore, climatic variables (temperature and relative humidity) were found to have an influence on the abundance of small mammals and fleas. List of Publication(s)/Conference(s) Proceedings Presentation of the thesis result at a small mammals conference in Arusha, Tanzania 1. Title: Prevalence of Leptospira and fleas in small mammals from Mukwe constituency, Kavango East Region of Namibia Presented by: Ms. Saima Kapia (University of Namibia) 2. Prevalence of haemoparasites on small mammals in Fallow land fields within Mukwe constituency, Kavango East Region, Namibia. Author of the presentation: Ms. Saima Kapia (University of Namibia) Presented by: Dr. Seth J. Eiseb (University of Namibia) Presentation of the thesis result at the Combined Congress of the Entomological and Zoological Societies of Southern Africa that was held at the CSIR International Convention Centre, Pretoria, South Africa 1. Title: Prevalence of haemoparasites on small mammals in fallow-lands within Mukwe Constituency, Kavango East Region, Namibia Author of the presentation: Ms. Saima Kapia (University of Namibia) Presented by: Dr. Seth J. Eiseb (University of Namibia) ii Table of Contents Abstract………………………………………………………………………………...i List of Publication(s)/Conference(s) Proceedings……………………………………ii Table of Contents………………………………………………..................................iii List of Tables…………………………………………………...…………………..viii List of Figures……………………………………………………………………….ixx List of Abbreviations and/or Acronyms…………………………………………......xii Acknowledgements………………………………………………………………….xiv Dedication……………………………………………………………………….....xxvi Declaration…………………………………………………………………...…….xviii Chapter 1………………………………………………………………………………1 1. Introduction…………………………………………………………………………1 1.1 Diversity of symbiotic interactions .......................................................................... 1 Mammals and associated parasites: complex interactions strongly influenced by biotic and abiotic parameters. ........................................................................................... 3 Pathogenic Leptospira ................................................................................................... 3 Fleas ............................................................................................................................... 5 Small mammals .............................................................................................................. 7 1.2 Statement of the problem ......................................................................................... 9 1.3 Objectives of the study........................................................................................... 11 1.3.1 Broad objective ............................................................................................ 11 1.3.2 Specific objectives ....................................................................................... 11 iii 1.4 Hypotheses of the study ......................................................................................... 12 1.5 Significance of the study ........................................................................................ 13 1.6 Limitations of the study ......................................................................................... 15 1.7 Delimitation of the study ....................................................................................... 16 Chapter 2……………………………………………………………………………..17 2. Literature Review………………………………………………………………….17 2.1 Leptospira .............................................................................................................. 17 2.1.1 General aspects of Leptospira ..................................................................... 17 2.1.2 Typical life cycle of pathogenic Leptospira ................................................ 18 2.1.3 Epidemiology and host association ............................................................. 19 2.2 Fleas ....................................................................................................................... 21 2.2.1 Flea morphology .......................................................................................... 21 2.2.2 The flea life cycle ........................................................................................ 22 2.3 Small mammal flea hosts ....................................................................................... 24 Chapter 3……………………………………………………………………………..30 3 Materials and methods……………………………………………………………..30 3.1 Map of the study site .............................................................................................. 30 3.2 Description of the study site .................................................................................. 31 3.3 Climate ................................................................................................................... 31 3.3.1 Rainfall ........................................................................................................ 31 3.3.2 Temperature, relative humidity and evaporation ......................................... 33 iv 3.4 Soils........................................................................................................................ 33 3.5 Flora, fauna and human livelihoods ....................................................................... 34 3.6 Field trapping of small mammals .......................................................................... 35 3.7 Weather data .......................................................................................................... 35 3.8. Laboratory processing ........................................................................................... 36 3.8.1 Flea identification ........................................................................................ 36 3.8.2 Small mammal collections and identification ............................................. 36 3.8.3 Detection of Leptospira ............................................................................... 38 Obtaining kidney samples .................................................................................... 38 Obtaining the DNA samples and Leptospira detection ........................................ 38 Leptospira identification..................................................................................... 399 3.9 Data analyses ......................................................................................................... 40 3.9.1 Leptospira .................................................................................................... 40 3.9.2 Fleas ............................................................................................................. 40 Abundance of fleas ............................................................................................... 40 Similarity index for fleas ...................................................................................... 41 Species diversity ..................................................................................................