THE EFFECT OF EXTENDED SPECTRUM BETA - LACTAMASES ON READY TO EAT MEAT, ‟ BEEF AND CHICKEN SUYAˮ.

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 P^H 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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