ABSTRACT

This study was undertaken to highlight the detection and susceptibility of some bacterial associated with quail eggs. Quail egg shells from some farms in Kaduna State were screened for incidence of Escherichia coli, Salmonella species and Pasturella species using standard microbiological procedure. The isolates were screened for their susceptibility to a panel of ten commonly prescribed antibiotics using disc diffusion method as described by Clinical Laboratory Standard Institute. Conjugation studies and curing experiments were also carried out on resistant bacteria to determine involvement of plasmid in the resistance. Plasmid DNA analysis was also done by using Agarose Gel Electrophoresis. Furthermore, PCR amplification of target resistance genes was also investigated. The results obtained from this study showed that the bacterial isolates i.e Escherichia coli, Salmonella species, Pasturella species from the three study sites were highly prevalent in the investigated quail egg shells. The result of this study indicates that resistance to Penicillin, Bacitracin, Tetracycline, Trimethoprim and erythromycin is relatively high among the farms investigated and their continued usefulness in the treatment of infections of E. coli, Salmonella species and Pasturella species needs be reviewed. All the isolates were found to habour the inck epidemiologic plasmid encoding gene for the blactx-m of extended beta lactamases and the gyrase B gene which code for resistance to quinolones. Plasmid borne resistant genes among bacterial test isolates were present in Mai Doki Agric Limited and Amir heritage farms Kaduna, Nigeria. There should be strict attention to infection control guidelines to learn the spread of resistant organisms when they develop i.e irrational use of antibiotics should be discouraged to restrict antibiotic use to bacterial infections since it encourages development of drug resistance.

CHAPTER ONE

INTRODUCTION 1.1 BACKGROUND OF STUDY
Bacteria such as E. coli, Salmonella spp and Pasturella spp have been reported to be the common causes of poultry food borne illnesses and death in humans (Ezekiel et al., 2011). Cross-contamination of foods by these organisms can occur coupled with the spread of genes resistant to commonly used antibiotics along various points in the processing line of sale of poultry products. This is of both serious public health and economic concerns. The symptoms of food borne illnesses, resulting from the consumption of pathogen contaminated foods, can range from mild to more severe indications such as diarrheoa, fever, nausea, vomiting, abdominal cramps, dehydration, meningitis, endocarditis, kidney failure and septicemia (Darwin and Miller, 1999).
The worldwide increase in the use of antibiotics in poultry and livestock production industry to treat and prevent infectious bacterial diseases and as growth promoters at sub-therapeutic levels in feeds has led to emergence of bacterial resistance to antibiotics during the past years (Apata, 2009). The increased use of antibiotics has been reported to play a significant role in the emergence of antibiotic resistant bacteria (Ashraf and Shah, 2011). Increasing episodes of multi-drug resistant pathogens can result in failure of antibiotic therapy in both animals and human. This can probably facilitate the transmission of antibiotic resistance between and among bacterial strains and species.There is an increase in public and government interest in phasing out inappropriate antibiotic use in animal husbandry (Cogan et al., 2001) because of the growing global concerns that antibiotic resistant bacteria can be transmitted from animals to humans.
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Improvement in the hygienic practice of handling raw animal products and adequate heat treatment to eliminate antibiotic resistant bacteria may play a role in preventing the spread. Recent scientific evidence has shown that resistance to antibiotics is not only due to the natural ability of a tiny fraction of the bacteria with unusual traits to survive antibiotic’s attack, enabling resistant strains to multiply, but also stems from the transmissibility of acquired resistance to their progeny and across to other unrelated bacterial species through extrachromosomal DNA fragment called the plasmids (WHO, 2011).
The emergence and spread of resistant bacterial strains like Campylobacter sp, and Enterococcus sp. from poultry products to consumers put humans at risk to new strains of bacteria that resist antibiotic treatment. Resistant bacteria thwart antibiotics by interfering with their mode of action via a range of effectors’ mechanisms, including synthesis of inactivating enzymes, alteration in the configuration of cell wall or ribosome and modification of membrane carrier systems. These mechanisms are specific to the type of resistance developed. Because of the growing global concerns that resistant bacteria can pass from animals to humans, there is an increase in public and governmental interest in phasing out inappropriate antibiotic use in animal husbandry. Improvement in the hygienic practice of handling raw animal products and adequate heat treatment to eliminate the possibility of antibiotic resistant bacteria surviving may play a role in preventing the spread. More attention should be focused on increasing antibiotic surveillance capacity to cope with the spread of emerging resistances and on the alternative approach to sub-therapeutic antibiotics in poultry, especially the use of probiotic micro-organisms that can positively influence poultry health and produce safe edible products.
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Escherichia coli, Salmonella species, and Pasteurella sp are the major bacterial pathogens isolated from poultry. Difference in susceptibility to antibiotics by these microorganisms has become a major factor in drug choice and success of treatment. Great concerns have been raised regarding emerging antimicrobial resistance among bacteria that may result in unpredictable antimicrobial susceptibility and failure of therapy (Apata, 2009). The primary objective of the present study therefore,is to determine the levels of antimicrobial susceptibility/resistance of E. coli, Salmonella species, and Pasturella speices isolated from quail egg shells. 1.2 STATEMENT OF RESEARCH PROBLEM/JUSTIFICATION OF STUDY Poultry feeds containing antimicrobial additives have been reported to enhance development of high levels of resistance especially among enteric organisms. Furthermore, the quail farmers and harvested eggs are prone to transmit these pathogenic resistant organisms to the local populace. 1. The result obtained would provide a basis on the raw consumption of quail eggs. 2. This finding will provide standard needed to be exploited before quail eggs can be consumed. 3. The results of this study will also display the common bacteria isolates and its susceptibilities in Kaduna town, Nigeria. 4. The prevalence bacterial resistance markers/genes will be highlighted in this study. 5. The result of this study will add to knowledge in the field of Epidemiology and Microbiology.
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1.3 AIM AND SPECIFIC OBJECTIVES 1.3.1 Aim The aim of this study is to isolate E.coli, Salmonella sp, and Pasturella sp contaminating quail egg shells and determine their susceptibility to commonly prescribed antibiotics towards improved management of poultry farms. 1.3.2 Objectives The specific objectives of this study include: 1. Isolation and identification of E.coli, Salmonella spp, and Pasturella spp from quail egg shells. 2. Determination of susceptibility of the isolated organisms to selected commonly prescribed antibiotics. 3. Determination of Minimum Inhibitory Concentration of selected antibiotics against resistant bacterial isolates. 4. Determination of resistance plasmid by conjugation, curing, and characterization. 5. Amplification of transferable resistance factor using gene specific primers. 1.4 HYPOTHESES 1.4.1 NULL HYPOTHESIS Quail eggs are not contaminated with bacteria, but if they do, they are not resistant to commonly prescribed antibiotics.
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1.4.2 ALTERNATE HYPOTHESIS Quail eggs are contaminated with bacteria and they are mostly resistant to commonly prescribed antibiotics.
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