STUDIES ON BRUCELLOSIS IN CATTLE IN JIGAWA STATE, NIGERIA


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TABLE OF CONTENTS
Titles
Abstract
List of Abbreviations

CHAPTER 1: INTRODUCTION
1.1       Background of the Study
1.2       Statement of Research Problem
1.3       Justification of the Study
1.4       Aim of the Study
1.5       Objectives of the Study
1.6       Research Questions

CHAPTER 2: LITERATURE REVIEW
2.1       History of cattle Brucellosis
2.2       The Genus Brucella
2.2.1    Classification scheme
2.2.2    Genomics
2.2.3    Proteomics
2.3       Epidemiology
2.3.1    Epidemiology of brucellosis in Europe, Middle East and North Africa
2.3.2    Epidemiology of brucellosis in Asia
2.3.3    Epidemiology of brucellosis in Latin America
2.3.4    Epidemiology of brucellosis sub-Saharan Africa
2.3.5    Situation of bovine brucellosis in Nigeria
2.4       Virulence and Pathogenicity
2.5       Immunity and Immune Mechanisms
2.6       Pathogenesis
2.7       Pathology
2.8       Diagnosis
2.8.1    Samples          
2.8.2    Tests for the Detection of Brucella
2.8.3    Direct microscopic examination
2.8.4    Isolation
2.8.5    Biochemical tests
2.8.6    Polymerase chain reaction
2.8.7    Serological Tests
2.8.7.1 ELISA
2.8.7.2 Rose bengal plate test
2.8.7.3 Serum agglutination test
2.8.7.4 Complement fixation test
2.8.7.5 Milk ring test
2.8.7.6 Whey agglutination test
2.8.8 Test to demonstrate an allergic reaction to Brucella
2.9  Molecular Epidemiology
2.9.1 Multi-locus variable number tandem repeat analysis
2.9.2 Outer membrane protein typing
2.9.3 IS711 Typing
2.9.4 Amplified fragment length polymorphism
2.9.5 Pulsed- Field gel electrophoresis
2.9.6 Multi locus sequence typing
2.10     Treatment
2.11   Control and Prevention of Brucellosis
2.12   Future Areas of Research in Brucellosis
2.12.1 Subunit vaccines
2.12.2 Immunomodulators
2.12.3 Serum components other than antibodies
2.12.4 Antibody therapy
2.12.5 Liposome encapsulated antibiotics and vaccines
2.12.6 Cross-reactions and cross-protection

CHAPTER THREE: MATERIALS AND METHODS
3.1       The Study Area
3.2       Research Materials
3.3       Sample Size Determination
3.4       Survey Design
3.5       Sample Collection
3.5.1 Herd samples
3.6       Serological test Procedure
3.6.1 Procedure for rose bengal plate test
3.6.2    Procedure for serum agglutination test
3.6.3    Procedure for complement fixation test
3.6.4    Procedure for milk ring test
3.6.5    Procedure for whey agglutination test
3.7   Culturing samples for Brucella Isolation
3.7.1 Media preparation
3.7.2 Culture of milk, swabs, tissue samples and stomach contents
3.8 Gram stain
3.9 Modified Ziehl-Neelsen stain
3.10     Biochemical Characterization Brucella Isolates
3.10.1  Growth in the presence of carbon dioxide
3.10.2  Urea hydrolysis
3.10.3  Hydrogen sulphide production
3.10.4  Agglutination with monospecific antisera A and M
3.10.5  Sensitivity to thionin and basic fuchsin dyes
3.10.6  Biotyping of Brucella isolates
3.11     Questionnaire Analysis
3.12     Data Analysis

CHAPTER FOUR: RESULTS
4.1       Samples Collected from the Three Senatorial Zones of Jigawa State Nigeria
4.2       Prevalence of Brucella Antibodies in Serum samples from the Three Senatorial Zones of Jigawa State Nigeria based on RBPT, SAT and CFT Tests
4.3       Degree of Agreement between Rose Bengal Plate Test, Serum Agglutination Test and Complement Fixation Test based on Kappa values
4.4       Overall Prevalence of Brucella Antibodies in Serum samples in Jigawa State based on the Screening tests (RBPT and SAT) and the Confirmatory test (CFT)
4.5       Individual Animal Prevalence (IAP) Based on RBPT, SAT and CFT in the North Central Senatorial Zone
4.6       Individual Animal Prevalence (IAP) Based on RBPT, SAT and CFT in the North East Senatorial Zone
4.7       Individual Animal Prevalence (IAP) Based on RBPT, SAT and CFT in the North West Senatorial Zone
4.8       Within Herd Prevalence (WHP) Based on RBPT, SAT and CFT in the North Central Senatorial Zone
4.9       Within Herd Prevalence (WHP) Based On RBPT SAT and CFT in the North East Senatorial Zone of Jigawa State
4.10     Within Herd Prevalence (WHP) Based on RBPT SAT and CFT in the North West Senatorial Zone
4.11     Overall Herd Prevalence of Brucella Infected Herds based on CFT in the Three Senatorial Zones of Jigawa State Nigeria
4.12     Prevalence of Brucella Antibodies in Milk in the 3 Senatorial Zones of Jigawa State based on Milk Ring Test (MRT)
4.13     Prevalence of Brucella Antibodies in Milk in the 3 Senatorial Zones of Jigawa State based on Whey Agglutination Test (WAT)
4.14     Seroprevalence of Brucella antibodies Cattle in Jigawa State Nigeria based on Age, Sex and Breed
4.15     Seropositivity among herds with absence or presence of brucellosis Clinical signs in the study areas
4.16     Isolation of Brucella Species from Milk, Swabs, Tissues and Stomach Contents
4.17     Identification of Brucella isolates
4.17.1  Gram staining and Modified Ziehl-Neelsen staining
4.17.2  Biochemical Tests
4.18     Questionnaire Analysis
4.18.1  Demography of herd owners
4.18.2  Herd information
4.18.3  Herd owners knowledge/awareness of brucellosis
4.18.4 Assessment of factors influencing the introduction of brucellosis into the study herds
4.18.5  Assessment of factors influencing the maintenance of brucellosis into the study herds
4.18.6 Assessment of factors influencing the spread of brucellosis within the study herds based on CFT

CHAPTER FIVE: DISCUSSION

CHAPTER SIX: CONCLUSIONS AND RECOMMENDATIONS
6.1       Conclusions
6.2       Recommendations
REFERENCES
APPENDICES




ABSTRACT

A serological and bacteriological study of Brucella infection in cattle was carried out in Jigawa State Nigeria in order to determine the seroprevalence, identify isolates and assess factors responsible for the introduction, maintenance and spread of the Brucella infection in cattle herds in selected Local Government Areas of Jigawa State. The study employed Rose Bengal plate test (RBPT) and Serum Agglutination test (SAT) as screening tests and Complement Fixation test (CFT) as a confirmatory test. Brucella organisms were isolated using cultural methods and biochemical tests were used to identify Brucella species. Milk Ring (MRT) and Whey Agglutination tests (WAT) were used to screen milk samples. Questionnaires were used to identify presence of exposure factors to Brucella species. From the results of the study, 93 (4.98%), 83(4.44%) and 68(3.64%) out of the 1867 serum samples were positive with RBPT, SAT and CFT tests respectively. There was a strong degree of agreement between the screening and the confirmatory tests as indicated by Kappa value of 0.926 0.846 and 0.827. Prevalence of

Brucella antibodies in the study herds indicated that 30 out of the 147 herds sampled were positive using RBPT, SAT and CFT tests, with an individual animal prevalence (IAP) ranging from 0 to 18.6% and a within herd prevalence (WHP) from 0 to 57.14% based on the CFT tests. The overall herd prevalence in the study based on the confirmatory test was 20.40 % (30/147). The results of Milk samples screened by MRT and WAT tests showed that 0.5% (5/869) and 0.72% (2/274) tested positive with the two different tests respectively. The results also showed that prevalence of Brucella infection was higher among female cattle 3.4% (70/2053) than males 0.52% (5/957) in all the study herds. The rate of seropositivity was highest among cattle of age group 37-72 months, 4.55% (37/813). Foreign breeds presented higher seropositivity of 3.2% (8/249) than the local breeds 2.4% (67/2746). A total of 900 milk, 460 vaginal swabs, 325 prepucial swabs, 14 placentae, 11 stomach contents, 11 spleen and 11 liver samples were cultured for Brucella isolation. Two isolates were obtained, one each from placenta and vaginal swabs of Friesian and Bunaji cattle in the North Central and North West Senatorial Zones. Biochemical characterization of the isolates indicated that they were both Brucella abortus. From the study all the herd owners interviewed were aware of brucellosis as a disease entity with the local name Bakkale, but they differed in ways of recognizing it. The results also showed that 66 % (98/147) of herd owners used a local herb “Madaci” for treatment of Brucella infection, while 33.3% (49/147) utilize veterinary services. The study also indicated that none of the herds were vaccinated against brucellosis. Assessment of exposure factors responsible for introducing, maintaining and spread of Brucella infection between and within herds in the study areas indicated that there was statistical association between migration, hygiene practices, raising multiple species, housing, extensive husbandry system and natural breeding using bulls with prevalence of Brucella infection (P< 0.05). In conclusion the isolation of two Brucella abortus species and evidence of Brucella infection in the study herds has demonstrated that Brucella exists among cattle in the State. It was hereby recommended that cattle owners should be educated on the various methods of preventing and controlling brucellosis in the State.



CHAPTER ONE

INTRODUCTION

1.1              Background of the study

Livestock plays a crucial role in the livelihood of the majority of Africans including the citizens of Nigeria (JARDA, 2010). It is one of the main sources of meat, milk, draught power and cash income. It also plays an important role in socio-cultural traditions of many African setting (McDermott and Arimi, 2002).


In Jigawa State, livestock and poultry are mainly kept by small to medium scale farmers in the semi-arid regions of the State (JRI, 2003). Livestock production sectors are defined in to three categories as small, ranging from 1 to 30 animals, medium 31 to 50 animals and large, more than 50 animals based on total number of cattle, goats and sheep (Livestock Committee Report, 2013).


Several documented bacterial diseases which are directly and indirectly transmitted occur commonly in Africa and are poorly controlled in both livestock and human populations (McDermott and Arimi, 2002; Smits and Cutler, 2004). These diseases such as brucellosis have a great socio-economic impact leading to low production due to abortion, reduced milk production, and loss of draft power, which ultimately have negative effects on cash income (Smits and Cutler, 2004). However, despite their economic and social importance, livestock management as well as programmes to control infectious diseases in Africa.....

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