ABSTRACT
The inappropriate use of antibiotic on poultry birds has made Staphylococcus aureus resistant to antibiotic which has become a problem in the medical sector. This study was aimed at evaluating the beta-lactamase production potentials of S. aureus isolates associated with various poultry birds. Swabs from the different birds studied (quail, pullet, Local birds, turkey and guinea fowl) were inoculated into Mannitol salt agar plate. A total of Eighty-two (82) S. aureus was recovered from a total of 100 birds sampled. A greater percentage of the S. aureus was obtained from turkey (100%) and guinea fowl (85%) respectively. The results of the antibiotic susceptibility profile revealed that the most effective antibiotic against the isolates was Oflaxacin with 100% susceptibility rate follow closely by gentamycin, recording 80% susceptibility. High level of resistance to Augmentin, Ceftazidime, Cloxacillin, Cefuroxime and Cefotaxime was recorded amongst the S. aureus isolates of the turkey S aureus isolates recovered from the birds, 57 (72.15%) were found positive to beta-lactamase production while 25 (31.64%) were negative to beta-lactamase production. The findings of this study therefore calls for proper caution to prevent transmission of antibiotic resistant S. aureus from poultry to humans which could further add to the already elevated antibiotic resistance problem.
TABLE OF CONTENTS
Table page i
Certification ii
Acknowledgements iii
Abstract iv
Table of
Contents vii
List of
Tables viii
CHAPTER ONE
INTRODUCTION
1.1 Background of study 1
1.2 Aim and objectives 4
CHAPTER TWO
LITERATURE REVIEW
2.1 Staphylococcus
aureus: characteristics and pathogenicity 5
2.2 Antibiotics resistance in poultry
birds 7
2.3 Antibiotic resistance 8
2.4 Causes of antibacterial resistance 9
2.5 Top line view of β-lactamases 9
2.6 β-lactamase origins 10
2.7 β-lactamases in gram-positive
bacteria 14
2.8 β-lactamases classification 16
2.9 Beta-lactamase-producing bacteria and
their role in infection 17
2.10 Beta-lactamase inhibitors 17
2.11 Beta-lactamase and beta-lactamase
inhibitors 18
CHAPTER THREE
3.1 Materials and methods 19
3.2 Sterilization of materials 19
3.3 Preparation of culture media 19
3.4 Isolation of antibiotics-resistant Staphylococcus aureus 19
3.5 Purification of isolates 19
3.6 Identification of the
antibiotics-resistant Staphylococcus
aureus 20
3.7 Gram staining 20
3.8 Biochemical test 20
3.8.1 Catalase Test 20
3.8.2 Coagulase Test 21
3.8.3 DNase Test 21
3.19 Antibiotics susceptibility testing 21
3.10 Test for beta-lactamase production 22
3.10.1 Beta-lactamase determination using acidimetric
method 22
CHAPTER FOUR
RESUITS 23
CHAPTER FIVE
DISCUSION, CONCLUSION AND RECOMMENDATION
5.1 Discussion 34
5.2 Conclusion 36
5.3 Recommendation 36
LIST
OF TABLES
Table Title Page
4.1 Frequency of isolation of S. aureus from poultry birds 27
4.2 Cultural Morphology and
Biochemical characteristics of the isolates 28
4.3 Beta-lactamase profile of S. aureus isolated from different
birds 35
LIST OF FIGURES
Figure Title Page
4.1 Antibiotic susceptibility profile of the
Quail bird isolates 29
4.2 Antibiotic susceptibility
profile of the Pullet bird isolates 30
4.3 Antibiotic susceptibility
profile of the Local bird isolates 31
4.4 Antibiotic susceptibility
profile of the Broiler isolates 32
4.5 Antibiotic susceptibility
profile of the Guinea fowl isolates
33
4.6 Antibiotic susceptibility
profile of the Turkey isolates 34
INTRODUCTION
1.1 Background of study
Staphylococcus aureus
is a normal nasal flora of humans and animals, although they are generally
considered commensal bacteria; they have the potential to cause a number of
infections which constitute health concerns for woman, new born, elderly, and
immune compromised individuals (Gardete and Tomasz, 2014). In human, Staphylococcus aureus has been
implicated in disease such as dermatitis, pneumonia, septicaemia, osteomyelitis
and meningitis in both humans and swine, as well as bovine mastitis in cattle
and bumble-foot disease in poultry (Quinn et
al., 2000) In poultry, the disease conditions associated with Staphylococcus vary with the site and route of inoculation in
hatchery and poultry farms, and can infect the bones, joints, tendon sheaths,
skin, sterna bursa navel, and yolk sac through breakage of the skin and mucosal
membrane of the birds, The immune compromised ones are often more prone to Staphylococcal infections. Once in the
host, Staphylococcus aureus invade the blood stream,
resulting in systemic infection in multiple organs, there by influencing
economic losses, which accrued as a result of weight loss, decreased egg
production, lameness, mortality, and condemnation at slaughter (Altahat et al., 2012). Reports have shown that
prevalence of enter toxigenic Staphylococcus
aureus in food handlers that serves as vehicle for zoonotic
dissemination of pathogenic Staphylococcus
aureus among poultry from workers,
communities and hospitals varies in industries and countries. In Japan, a
retail survey performed between 2002 and 2003 found 17.6% of raw chicken meat
infested with enter toxigenic Staphylococcus
aureus; an indication of future and possible Staphylococcosis outbreak, which could influence increased
mortality and morbidity. To control and manage these disease/pathogenic Staphylococcus aureus, inappropriate use
of antibiotics is employed in both poultry farms and clinical setting. In
poultry management, antibiotics are often used in animal food production for
growth promotion and routine disease prevention without prescription or control
measures. This has necessitated the development of drug resistant superbug such
as methicillin resistant Staphylococcus
aureus (MRSA), vancomycin intermediate Staphylococcus
aureus (VISA), and vancomycin resistant Staphylococcus
aureus (VRSA), which are now known as major emerging public health problem
(Bala et al., 2016). With increase in
population density within a particular geographical location, the incidence of
both community associated and hospital associated multidrug resistant Staphylococcus aureus have been observed
to increase with time, regardless of hospitals size and control measures due to
drug abuse and zoonotic transfer of resistance gene mainly located on mobile
element, such as plasmids or prophages and transferable through horizontal gene
transfer (Igwe et al., 2013).
In
Nigeria, the antibiotics use by poultry farmers from various regions in Nigeria
varied among studies. Among the most commonly used agents reported in
literature include neomycin and gentamycin (Van et al., 2007) enrofloxacin and chlortetracycline (Adelowo et al.,
2009), tetracycline and sulphonamide (Nahar et
al., 2014) fluoroquinoloes (Oluwasile et
al., 2014), as well as gentamycin and tetracycline (Adebowale et al., 2016). Based on responses
received from respondents, it is evident that most of the poultry famers do not
obtain information on antibiotics they use from qualified personnel.
Staphylococcus aureus is a major pathogen that can cause various
forms of diseases varying from simple to life-threatening infection in human
population (lowy 1998, Diekema et al.,
2001). The invasion of the host tissue by Staphylococcus
aureus apparently involves the production of a formidable array of
extracellular enzymes (invasion) which facilitates the invasive process. Some
may occur also as cell associated proteins by breaking down primary and
secondary defences of the host which can facilitate the growth and spread the pathogen.
The damage of the host as a result of this invasive activity may become part of
the pathology of an infection (Noble, 2010).
Beta-lactam compounds such as
penicillin continues to be one of the most frequently used drugs in veterinary
medicines (Pitkala et al., 2007). Two
primary resistance mechanisms to beta-lactams are noteworthy of Staphylococcus spp. The expression of
beta-lactamase enzymes encoded by the blaz gene, and production of the
penicillin-binding protein 2a (Pbp2a), resulting in a higher-level of
resistance encoded by the mecA gene (Fuda et
al., 2005). Prevalence of penicillin resistance in Staphylococci causing animal disease is most commonly due to the
blaz gene (Pitkala et al., 2007).
Beta-lactam
antibiotics are among the most frequently prescribed antibiotics worldwide in
the control of Staphylococcus-aureus
infection. They act on peptidoglycan synthesis by molecularly acting on
transpeptidase and carboxypeptidase thereby disrupting cell wall formation of
the pathogen. However, the efficacy of antibiotics for therapy have suffered a
setback due to growing trend of multidrug resistant strains observed in the
organism to Beta-lactam and other antibiotics (Lowy et al.,2003, Deurenberg and Stobberingh, 2008).
Different
test can be performed to evaluate beta-lactamase production in Staphylococci. A qualitative procedure
for detecting production of beta-lactamase is the usage of Nitrocefin disks.
The reaction is based on the production of a coloured compound when the
substrate (nitrocefin) is exposed to a beta-lactamase-producing- bacteria. The
clover leaf test (CLT) is an alternative with high sensitivity and specificity
for investigating beta-lactamase production in staphylococci (Bergan et al., 2007).
In addition extended spectrum beta-lactamase
(ESBL.) capable of hydrolyzig penicillins, broad spectrum cephalosporins and
monobactams in enterobacteriaceae (David and Bonomo, 2005) are often located on
plasmids that are transferable from strain to strain and between bacteria
species (Rupp and Fey, 2003). Extended spectrum beta-lactamase (ESBL) producers
have continued to draw attention globally with their attendant clinical failure
to new generation antibiotics and nosocomial spread (Olowe et al., 2012). In addition rapid change over time in extended
spectrum beta-lactamase has been observed with variations within geographic
areas. Clinical outcomes data indicates that extended spectrum beta-lactamase
are clinically significant and when detected, suggest the need for the use of
appropriate antibacterial agents. Hence, this study was designed to isolate
antibiotics resistant Staphylococcus
aureus from poultry birds in Michael Okpara University of Agriculture,
Umudike, and also evaluate B-lactamase production by the antibiotics resistant Staphylococcus aureus isolate.
1.2 AIM AND OBJECTIVES
The
aim of this study was to isolate antibiotic resistant Staphylococcus aureus from poultry birds in Michael Okpara
University Of agriculture, Umudike, and also evaluate Beta-lactamase production
by the Staphylococcus aureus isolated
while the specific objectives were;
1.
To isolate and identify Staphylococcus aureus from the swabbed
samples of different
Poultry birds.
3. To
determine the antimicrobial susceptibility profile of the isolate from the
swabbed sample.
4. To screen the Staphylococcus aureus isolate for beta lactamase production.
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