ABSTRACT
In the present study,10 samples of beef suya and chicken suya was collected respctively from different food vendors at different strategic location in Ikwanuo LGA , Abia State were analyzed for screening of Extended spectrum Beta- Lactamase producing organisms on the ready to eat food; beef and chicken suya. Organisms isolated where Escherichia coli, Klebsiella pneumoniae, Salmonella spp and Citrobacter spp; using different cultural media and biochemical test; Gram staining reaction/Microscopy. Antimicrobial susceptibility test were carried out using 12 antibiotics discs; Organisms isolated were susceptible to the following antibiotics: Amoxicillin/Clavulanate, Imipenem/Cilastatin, Ofloxacin, Gentamicin, Levofloxacin. Confirmatory test (Double Synergy Test) for ESBL Production was conducted, some bacterial strains isolated were positive while some were negative. These finding shows that retailed foods especially ready to eat foods are contaminated with microorganisms due to improper handling of foods.
TABLE
OF CONTENT
Cover
page i
Title
Page ii
Certification iii
Dedication iv
Acknowledgements v
Table
of Contents vi
List of
Tables viii
List of
Figures ix
Abstract x
CHAPTER ONE: INTRODUCTION
1.0 Background of study 1
1.1
Objectives of the Study 4
CHAPTER TWO: LITERATURE REVIEW
2.0 Suya 5
2.1 Shelf life of Chicken and Beef Suya 6
2.2 Impact of Microbiological Spoilage of
Beef and Chicken Suya 7
2.3 Sources of Microbial Contamination 7
2.4 Microbiological Spoilage Mechanism in
Beef and Chicken Suya
8
2.5 Prevention and Control of Microbial
Spoilage 9
2.6 Method for Detection and Isolation of
Organism that causes Spoilage on Suya
10
2.7 Microbiological Spoilage Defect of Beef
and Chicken Suya 11
2.8 Bacterial Pathogens Association with Beef
and Chicken Contamination 12
2.8.1 Citrobacter
spp 12
2.8.2 Pathogenic Escherichia coli 12
2.8.3 Salmonella
spp 13
2.8.4 Shigella spp 14
2.8.5 Enterobacter spp 15
2.8.6 Klebsiella
pneumoniae
15
2.8.7 Staphylococcus aureus 16
2.8.8 Yersinia spp
16
2.9
Beta-Lactamase
17
2.10
Extended Spectrum Beta-Lactamases (ESBL) 18
2.11
Methods of Detection Extended Spectrum Beta – Lactamase 20
2.11.1
Screening Test 21
2.11.2
Confirmatory Test
21
CHAPTER THREE: MATERIALS AND METHODS
3.0 Study Area
23
3.1 Collection
of Samples
23
3.2 Materials,
Methods and Reagents 23
3.3 Sterilization of Materials 24
3.4 Preparation of culture media 24
3.5 Isolation
procedure
24
3.6 Purification
of the Isolates 25
3.7 Identification
of Isolated Organism 25
3.7.1 Gram
staining 25
3.7.2 Biochemical
Test 26
3.7.3 Indole
Test
26
3.7.4 Citrate
Utilization Test 27
3.7.5 Oxidase
Test 27
3.7.6 Coagulase
Test. 27
3.7.7 Hydrogen
Sulphide (H2S Production Test) 28
3.7.8 Starch
Hydrolysis
28
3.7.9 Motility,
Indole, Urease (MIU)
28
3.8 Antibiotics
Susceptibility Test
29
3.9 Confirmatory Test for ESBL-Producing
Organism
29
CHAPTER FOUR: RESULT 30
CHAPTER
FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATION
5.0 Discussion 47
5.1 Conclusion 51
5.2 Recommendation 57
REFERENCES 52
APPENDIX
TABLE
OF CONTENTS
Table 1: Viable microbial counts of retailed
beef suya sold in Ikwuano 32
Table 2: Morphology and Biochemical
identification of bacteria isolated from
retailed beef and chicken suya 33
Table 3: Percentage occurrence of Bacteria
isolated from retailed beef suya 34
Table 4: Antibiotics susceptibility pattern for
Escherichia coli isolated from retailed
Beef suya. 35
Table 4.1: Antibiotics susceptibility pattern for Escherichia coli isolated from retailed
Chicken Suya 36
Table 5: Antibiotics susceptibility pattern for
Salmonella spp isolated from retailed
Beef Suya sample 37
Table 5.1: Antibiotics susceptibility pattern for Salmonella spp isolated from retailed
Chicken Suya sample 38
Table 6: Antibiotics susceptibility pattern for
Klebsiella spp isolated from retailed
Beef
Suya sample 39
Table 6.1: Antibiotics susceptibility pattern for Klebsiella spp isolated from retailed
chicken suya sample 40
Table 7: Antibiotics
susceptibility pattern for Citrobacter
spp isolated from retailed
Beef Suya sample 41
Table 7.1: Antibiotics susceptibility pattern for Citrobacter spp isolated from retailed
Chicken Suya sample 42
Table 8: Antibiotics
susceptibility and Resistant pattern of all isolates from retailed
Beef Suya 43
Table 9: Antibiotics
susceptibility and Resistant pattern of all isolates from retailed
Chicken Suya 44
Table 10: Occurrence
of ESBLs among Bacterial isolates screened based on CLSI break
points. 45
Table 11: Prevalence
of
- Lactamase among Bacterial isolates
using double disk
synergy test (DDST) 46
CHAPTER ONE
1.0 Introduction
Ready to eat foods are foods
substances ingested or consumed by humans for the sustenance of life. However,
foods consumed by humans and animals serve as favorable media for rapid growth
of microorganisms because of the abundant organic matter, moisture and suitable
PH contained and because of the fact that the microorganisms are
equally living organisms. Presence of these microorganisms in foods warrants
their rapid multiplication which render the food unfit and dangerous for human
consumption (Yusha’u et al., 2010).
Processed meat products are defined
as those in which the properties of fresh meat must have been modified by the
use of one or more procedure such as grinding, addition of seasoning agents,
alteration of color or heat treatment. Processed meat products in Nigeria
comprise ‘Tsire’ or ‘Suya’ ‘Kilishi’ and ‘Balangu’. ‘Suya’ is a popular
Nigerian traditional processed ready to eat roasted meat product. It is served
or sold in public place, along streets, in club houses, restaurants, Picnics and
homes. It is prepared from boneless meat animals such as muttons, beef or goat.(Son,
1995)
Chicken and beef suya are common
delicacies to many Nigerians. Chicken and beef Suya Vendors are found in almost
every neighborhood with a dense population for various daily formal or informal
economic activities. Suya is a street processed roasted and vended meat product
(Son, 1996).
The meat is trimmed from associated
connective tissues, nerves and vessels. The meat is artfully sliced into very
thin continuous sheet which are then cut into pieces. The pieces of meat are
staked on sticks, spiced with groundnut power and flour, salt vegetable oil and
flavorings such as monosodium glutamate or others. The sticks are then arranged
round the heat place for the meat to roast and the duration of roasting depends
on such factors as fire intensity and size of meat.
In Nigeria sale of beef suya first
started before that of chicken which emerged just recently and is not as wide
spread as the former. These foods are brought and eaten without further
processing or cooking. Street vending of foods is a common characteristic of
countries with high unemployment, low salaries and poor social security
programme (Bryan et al; 1998).
The traditional roasting of ‘Suya’ is
usually done by wood charcoal. The prepared ‘suya’ when being sold are usually
packaged in leftovers newspapers and sometimes in cellophane or nylon bags. Most
of the stages of ‘Suya’ preparation, materials used, packaging, the handlers
and the surrounding environment can serve as source of contaminants to the meat
products. However, consumers of street vended meat are little aware of the high
health risks they face. Street foods are exposed to various forms of
contamination at every stage of handling. Etok (1998) identified inefficient
roasting/heating duration, uneven temperature distribution and exposure to
unhygienic environments are crucial factor of infection and contamination. It
is the aim of the vendors to minimize shrinkage of the meat during roasting to
maximize profit but at the same time satisfy the demand and appetite of the
buyers. So the food is usually prepared in a rush when the buyers are in a
rush, or they are roosted, kept exposed and cold to await would be consumers.
Whichever way, the suya would not be properly roasted or kept under safe
condition for consumption.
The Enterobactericeae are major pathogens in animals as well as humans
many strains of E.coli are harmless
and are found naturally in the gut of humans and animals. Traditionally, it
presence in foods has been an indication of faecal contamination of food or
water. However, particular strains are pathogenic travelers’ diarrhea and
hemolytic uraemic syndrome (HUS) are caused by some E.coli strains. The pathogenic types are said to be rare but in the
last few years, there have been several food borne out breaks from certain
strains of E.coli.
ESBLs-
Extended Spectrum Beta-Lactamase were first described in 1980s and have been
detected in Kiebsiella spp.and later
in Escherichia coli, Pseudomonas aeruginosa and Serratia marcesensand other gram
negative bacilli. ESBLs are enzymes comparing broad resistance to Penicillins,
Aztreonam and Cephalosporins and Monobactams. They are an increasing important
cause of transferable multi drug resistance in gram negative bacteria
throughout the world. In fact, one of the important features of ESBL producing
strains is resistant to multiple clinically important antibiotics.
Great
attention has been paid to bacterial resistance to antibiotics in both human
and animal populations for adverse impact on morbidity and mortality form
diseases caused by resistant bacteria economic cost of therapy and real risks
of the spread of resistant strains among humans and animals. Emerging
antibiotics resistant E. coli strains
have been increasingly reported in many developing countries and is causing serious
public health concerns E. coli has
been recognized as a reservoir for resistant genes and can occupy multiple
niches in human and animal hosts. Acquired resistance of B-lactam antibodies in
mainly mediated by extended spectrum B-lactamases [ESBLs].
More has been focused on hospitals as
the primary reservoir and place of transmission of many antimicrobial –
resistant organisms there is need to shift interest to the role of Non-hospital
community, such as foods as a significant reservoir of resistant pathogens.
1.1 Aim and Objectives
The project therefore seeks to:
a.
Isolate, characterize and identify
the organism associated with Beef and Chicken suya.
b. Determine
the total bacteria count and coliform counts in different suya samples [beef
and chicken suya] from different locations.
c.
Determine the antibiotics
susceptibility pattern of the (organisms) isolates.
d.
To determine the ESBL producing
isolates.
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