ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF MICROORGANISMS ISOLATED FROM READY TO EAT FOODS

ABSTRACT

Food borne disease outbreaks linked with Ready to eat foods have been associated with various foodborne pathogens .The initial microbiological load on Ready to eat food ingredients is important, however, factors such as handling, processing, storage and display may influence the microbiological load of Ready to eat foods at the point of sale. The antimicrobial susceptibility  pattern of microorganisms isolated from ready- to eat- foods sold in Umuahia Metropolis for analysis, Ten (10g)  of food sample was  blended and homogenized in 90 mL sterile distilled water (10-1 dilution). Further tenfold serial dilution of the resultant homogenates was made to obtain 10-3 dilution respectively. From this dilution, aliquots of 0.1 mL was inoculated in replicate plates of different media using the spread plate technique .Nutrient agar (Oxoid) was inoculated for total aerobic count and Eosin Methylene Blue agar was used as a selective medium for the growth of the coliform bacteria and the mates were incubated at 37°C for 24-48 h. Colonies were counted at the end of the incubation period and expressed as Log CFU/g. Pure cultures of isolates were obtained by repeated subculture on nutrient agar and was  stored on slants at 4°C until characterized. Bacterial isolates was identified based on colonial morphology, microscopy and biochemical tests. The Ready to eat food samples used in this study were five (5) samples purchased from five different locations. The total viable count of bacterial population in all Ready to eat food samples were in the range of 1.43x106 to 4.5x106cfu/g. The total coliform count ranged from 0.9 x105 to 6.7 x 105 . The frequency of occurrences of bacteria isolated Ready to eat food samples showed a total of Sixteen (16) bacteria isolates belonging to five genera were obtained from Ready to eat food samples and identified as Staphylococcus spp., Escherichia coli, Proteus spp., Bacillus spp., and Klebsiella spp., based on their biochemical reaction. It showed that Proteus spp. 5 (31.3%) was most predominant bacterial isolates associated with Ready to eat food. This was followed by Escherichia coli 3 (18.8%), Bacillus spp. 3 (18.8%) and Staphylococcus aureus 3 (18.8%) Klebsiella spp. 2 (12.5%), was least predominant. The antibiotics susceptibility profile of the bacteria isolates showed that E.coli 28mm was more susceptible to the antibiotic drugs tested than every other isolates. In conclusion, this study presented the level of microbial spoilage and contamination status of ready to eat food meat sold in different restaurants in Umuahia, Abia State and the role of such contaminated meat in causing food poisoning amongst other foodborne related illnesses.

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgments                                                                                                                  iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          vii

Abstract                                                                                                                                  viii

CHAPTER ONE

•         INTRODUCTION                                                                                                  1

1.1 Aim and Objectives                                                                                                          2

1.2 Objectives                                                                                                                         2

CHAPTER TWO

2.0   LITERATURE REVIEW                                                                                            4

2.2       Food Hygiene                                                                                                             8

2.3       Factors that Contribute to Food –Borne Illness                                                          9

2.4       Features of Common Food –Borne Bacteria Pathogens                                            11

CHAPTER THREE

3.0 MATERIALS AND METHODS                                                                                   15

3.1 Sample Collection                                                                                                            15

3.2 Media To Be Used And It’s Preparation                                                                                      15

3.3 Sterilization                                                                                                                      15

3.4 Sample Preparation                                                                                                           15

3.6 Identification of Bacteria Isolates                                                                                     16

3.7 Characterization of Isolates                                                                                              16

3.7.1 Gram staining                                                                                                                16

3.7.2 Motility Test                                                                                                                  17

3.8 Biochemical and Cultural Characteristics                                                                        17

3.8.1 Catalase test                                                                                                                   17

3.8.2 Coagulase Test                                                                                                               17

3.8.3  Citrate Test                                                                                                                   18

3.8.4 Indole Test                                                                                                                     18

3.8.5 Triple Sugar Iron Test                                                                                                   18

3.8.6 Oxidase Test                                                                                                                  19

3.8.7  Urease Test                                                                                                                   19

3.9. Antibiotic Sensitivity Test                                                                                               19

CHAPTER FOUR

4.0 RESULTS                                                                                                                        21

CHAPTER FVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATION                                      27

5.1 Discussion                                                                                                                         27

5.2 Conclusion                                                                                                                        29

5.3 Recommendation                                                                                                              29

LIST OF TABLES

Table                                         Title                                Page

•                                                    Total Viable Count of Bacteria from Ready to eat food                 22

•                                                                     Total Coliform Count of Bacteria from Ready to eat food             23

•                                                                     Percentage of Occurrence of Bacterial isolates from Ready           24

to eat food

•                                                                     Morphology and Biochemical Identification of isolates                  25

•                                                                     Antibiotics Susceptibility Profile of the bacteria isolated    26

CHAPTER ONE

1.0  INTRODUCTION

Ready-to-eat (RTE) foods have been described differently by various organizations and individuals. The food and Agriculture Organization defines street foods as RTE foods and beverages prepared and/or sold by vendors and hawkers, especially in streets and other similar public places (Rane, 2011). RTE foods could be raw or cooked, hot or cold and can be consumed without further heat treatment (Rane, 2009). RTE food also could be fruits bought directly from street vendors or hawkers or at local markets and eaten immediately i.e. without necessarily having to cut , peel or rinse them before consumption as RTE foods have already been prepared by the vendors (FA0, 2008). Some RTE foods such as meat pie, fried fish, egg roll, etc. are regarded as potentially hazardous because such foods can support the growth of pathogens (FAO, 2009).

Such foods must be kept at certain temperatures and conditions to minimize the survival and growth of foodborne microorganisms and pathogens. Studies have revealed the frequent contamination of street food in many developing world including Nigeria (WHO, 2008). The consumption of street foods has been suggested to potentially increase the risk of foodborne diseases as street foods are readily contaminated from different sources as there is no proper safety, quality and hygiene practices during preparation . In fact, street foods have often been associated with travellers’ diarrhea and other foodborne diseases. Mensah et al. (2000) reported the presence of Bacillus cereus, Staphylococcus aureus, Shigella sonnei, Escherichia coli, and Salmonella arizonae on different foods sold on streets of Accra.  Nyenje et al. (2010) investigated the microbiological quality of readyto- eat foods sold in Alice, South Africa and reported the contamination of these foods by Listeria spp., Enterobacter spp., Aeromonas hydrophila , Klebsiella oxytoca , Proteus mirabilis, Staphylococcus aureus and Pseudomonas luteola. Study on the microbial safety of ready-to-eat foods; meat pie, beef sausage roll and egg roll, pealed orange, walnut and apple vended on highways; Onitsha-Owerri, South east, Nigeria, revealed the contamination of these foods by pathogens which include; Salmonella spp., S. aureus, E. coli, B. cereus, Shigella spp., Enterococci spp., A. niger and Pseudomonas spp. Nwachukwu and Nwaigwe (2010) and Ossa (2009) reported contamination of street foods by pathogens including Salmonella spp., Shigella spp., E. coli, etc. in different parts of Nigeria. Ready-to-eat (RTE) foods refer to foods that do not require further significant preparation other than reheating or completion of a cooking process (FEHD, 2001; FSAI, 2001). It has been reported that Ready to eat foods accounts for a large volume of sales of the food service sector, representing more than a third of the food service volume outputs (Powers and Barrow,1999). Food borne disease outbreaks linked with Ready to eat foods have been associated with various foodborne pathogens (Gilbreth et al., 2005; Gibbons et al., 2006).The initial microbiological load on Ready to eat food ingredients is important, however, factors such as handling, processing, storage and display may influence the microbiological load of Ready to eat foods at the point of sale (Beuchat and Ryu, 1997; Angelidis et al., 2006).

1.1 AIM AND OBJECTIVES

The aim of this study is to examine the antimicrobial susceptibility  pattern of microorganisms isolated from ready- to eat- foods sold in Umuahia Metropolis.

1.2 OBJECTIVES

1.     To isolate and identify microorganisms isolated from ready to eat foods sold in Umuahia, Metropolis.

2.     To characterize microorganisms associated with ready to eat foods sold in Umuahia, Metropolis.

3.     To determine the antimicrobial susceptibility profile of microorganisms associated with ready to eat foods sold in Umuahia, Metropolis.

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