Full Project – CHARACTERIZATION OF TRYPANOSOMES ISOLATED FROM BOVINE AND CANINE USING POLYMERASE CHAIN REACTION

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TABLE OF CONTENTS

Title Page

Certification

Dedication

Acknowledgements

Abstract

Table of Contents

List of Tables

List of Figures

CHAPTER ONE

1.0 Introduction

1.1 African Animal Trypanosomiasis

1.2       Statement of Problem

1.3       Justification of the Study

1.4       Aim and Objectives of the Study

CHAPTER TWO

2.0 Literature Review

2.1       Pre-History

2.2       Early History

2.3       Geographical Distribution

2.4       Classification of Trypanosomes

2.4.1 Classification of Trypanosomes of Economic, Medicinal and Veterinary Importance

2.5       Trypanosome Life Cycle

2.5.1 Life Cycle of the Vector

2.5.2 Forms of Human African Trypanosomiasis

2.5.3 Animal Trypanosomiasis

2.5.4 Host range

2.6       Prevention

2.6.1 Transmission

2.6.2 Pathogenesis

2.6.3Clinical Signs

2.6.4 Epidemiology

2.7       Molecular Biology of Trypanosomes

2.7.1 Nucleus

2.7.2 Kinetoplast DNA

2.7.3 Maxicircles

2.7.4 Minicircles

2.7.5 Genotyping of Trypanosome Parasites

2.8       Diagnosis of Animal Trypanosomiasis

2.8.1 Diagnosis

2.8.2 Clinical

2.8.3 Microscopic Diagnosis

2.8.4 Serological Diagnosis

2.8.5 Molecular Diagnosis

2.9       Control Measure of the Vectors

CHAPTER THREE

3.0 Materials and Methods

3.1       Study Site

3.2       Sample Site

3.3       Collection of Blood Sample

3.4       Preparation of Buffy Coat

3.4.1 PCV Measurement

3.5       Preparation of Thin Blood Smears

3.6       Preparation of Buffers

3.6.1 Preparation of Buffer AL

3.6.2 Preparation of Buffer AW1

3.6.3 Preparation of Buffer AW2

3.6.4 Buffer AE

3.7       DNA Extraction

3.8       PCR Analysis of the Genomic DNA

3.9       PCR Amplification

3.9.1 Agarose Gel Electrophoresis

CHAPTER FOUR

4.0 Results

4.1       Buffy Coat Examination

4.2       Thin Blood Smear Examination

4.3       Blood Parameters

4.4 Identification of Trypanosomes

CHAPTER FIVE

Discussion and Conclusion

References

ABSTRACT

Trypanosomiasis is a vector-borne parasitic disease, transmitted to animals by tsetse fly (Glossina genus) bites. The human type causes sleeping sickness in man while animal type causes nagana in animals. The aim of this study is to characterize trypanosomes isolated from dogs and cattle in Jos South Abattoir. Isolated trypanosomes were subjected to polymerase chain reaction using kin primers targeting internal transcribed spacer 1 (ITS1) of ribosomal deoxyribonucleic acids. The PCR allowed detection of three trypanosome livestock species namely T. vivax sub-species (150bp, 250bps); T. congolense (750bp) and T. brucei (540bp). The samples from dogs all showed T. vivaxsub-species and one sample showed mixed infection of T. vivax sub-species and T. congolense. The samples from cattle showed mostly mixed infections of T. brucei and T. congolense. The most significant finding is the observation that more than one sub-species of T.vivax is circulating in the Jos area.

CHAPTER ONE

1.0              Introduction

1.1              African Animal Trypanosomiasis

African animal trypanosomiasis, also known as sleeping sickness, is a vector-borne parasitic disease. The parasites concerned are protozoa belonging to theTrypanosoma genus. They are transmitted to humans by tsetse fly (Glossina genus) bites which have acquired their infection from human beings or from animals harbouring the human pathogenic parasite (WHO, 2014)

Tsetse flies are found just in sub-Saharan Africa though only certain species transmit the disease. For reasons that are so far unexplained, there are many regions where tsetse flies are found, but sleeping sickness does not exist .Rural populations living in regions where transmission occurs and which depend on agriculture, fishing, animal husbandry or hunting is the most exposed to the tsetse fly and therefore to the disease. The disease develops in areas ranging from a single village to an entire region. Within an infected area, the intensity of the disease can vary from one village to the other.The tsetse fly is about the same size as a honeybee. They are quite aggressive and the bite hurts (WHO, 2014).

African animal trypanosomiasis is a complex debilitating protozoan disease of animals ranked among the top 10 cattle diseases. (Perry et al., 2002).This group of diseases caused by protozoa of the genus Trypanosoma affects all domestic animals. The major species are T. congolense, T. vivax, T. brucei brucei and T. simiae. According to the Merck veterinary manual, T. congolense and T. brucei brucei are most common in the tsetse region of Africa;

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