ANTIMICROBIAL SUSCEPTIBILITY PATTERN OF MICROORGANISMS FOUND ON READY TO EAT FRUITS AND VEGETABLES

ABSTRACT

The study titled "Antimicrobial Susceptibility Pattern Of Microorganisms Found On Ready To Eat Fruits And Vegetables" aims to investigate the antimicrobial susceptibility patterns of microorganisms present on ready-to-eat fruits and vegetables within the vicinity of Michael Okpara Federal University of Agriculture, Umudike, Abia State. This study's objectives include isolating and characterizing bacteria present in these foods and determining their susceptibility to various antibiotics. Five different samples of ready-to-eat fruits and vegetables were collected aseptically from three different street vendors at the university gate, transported to the laboratory in insulated containers, and analyzed within 24 hours. Culture media were prepared by sterilizing all necessary equipment and using nutrient agar, MacConkey agar, Eosine methylene blue agar, and Salmonella-Shigella agar. The samples were then subjected to serial dilution, inoculated using the pour plate method, and incubated. Bacteria were isolated from these cultures and identified through cultural, microscopic, and biochemical analyses, including Gram staining, catalase, coagulase, citrate, oxidase, and motility tests. The antimicrobial susceptibility of the isolates was tested using the disc diffusion method on Muller-Hinton agar, following the guidelines of the Clinical and Laboratory Standards Institute. The study found a range of microbial loads on the fruits and vegetables, with six different bacterial species isolated: Staphylococcus aureus, Micrococcus, Klebsiella, Salmonella, Citrobacter, and Proteus mirabilis. These findings align with previous research, suggesting that the contamination may result from environmental factors, handling, and washing with contaminated water. The antimicrobial susceptibility tests indicated varying degrees of sensitivity and resistance among the bacteria to different antibiotics. The study highlights the need for improved hygiene practices among street vendors and increased consumer awareness to reduce the risk of foodborne illnesses. The presence of potential pathogens and antibiotic-resistant bacteria on ready-to-eat produce underscores the urgency of addressing food safety issues through better infrastructure, law enforcement, and education on safe food handling practices.

TABLE OF CONTENTS

Title Page                                                                                                     i

Certification                                                                                               ii

Declaration                                                                                                iii

Dedication                                                                                                 iv

Acknowledgements                                                                                    v

Table of Contents                                                                                      vi

List of Tables                                                                                            ix

Abstract                                                                                                       x

CHAPTER ONE: INTRODUCTION

1.1       Nutritional Value of Ready to Eat Fruits and Vegetables              2

1.2       Rate of Consumption of Ready to Eat Fruits and Vegetables        2

1.3       Contamination of Fresh Fruits and Vegetables                              2

1.4       Microbial Infection and Fresh Fruit and Vegetables                      3

1.5       Use of Antimicrobials in the Control of Pathogens                       3

1.6       Aims and Objective of this Research Work                                   3

CHAPTER TWO: LITERATURE REVIEW

2.0       What are Ready to Eat Fruits and Vegetables?                              4

2.1       Ready to Eat Fruits and Vegetables as a Source Of Food Borne Illness  4

2.2       Sources of Contamination of Ready to Eat Fruits and Vegetables 5

2.3       Food Contamination with Antibiotic Resistant Bacteria                5

2.4       Organisms of Interest in Ready to Eat Fruits and Vegetables        5

2.5       Microbial Quality of Ready to Eat Salad                                       6

2.6       Food Poisoning Outbreaks Caused By Ready to Eat Fresh Salad Vegetables                                                                                                  6

2.7       Decontamination Method Used by Produce Industry                    7

2.8       Review of Organisms Found On Ready to Eat Fruits and Vegetables                                                                                                  7

2.9       Microbiological Spoilage of Fresh Whole Fruits and Vegetables 9

2.9.1    Unusual Characteristics of Spoilage Micro Organisms               10

2.9.2    Prevention and Control Measures                                                14

CHAPTER THREE:  MATERIALS AND METHODS

3.1       Study Area                                                                                    17

3.2       Collection of the Sample / Specimen                                           18

3.2.1    Preparation of Culture Media                                                       18

3.2.2    Procedure for Preparation of Diluent Used                                  18

3.3       Bacteria Inoculation                                                                     19

3.4       Enumeration of Bacteria                                                              19

3.5       Isolation of Bacteria                                                                     20

3.6       Identification and Characterization of Isolates                             20

3.6.1    Microscopic Examination / Gram Staining Technique                20

3.6.2    Biochemical Reaction Test                                                           20

3.6.2.1 Catalase Test                                                                                 21

3.6.2.2 Coagulase Test                                                                              21

3.6.2.3 Citrate Test                                                                                   21

3.6.3.4 Oxidase Test                                                                                 21

3.6.3.5 Motility Test                                                                                 22

3.7       Antibiotic Susceptibility Tests                                                     22

CHAPTER FOUR:  RESULT

4.1       Result                                                                                            23

CHAPTER FIVE: DISCUSSION AND CONCLUSION

5.1       Discussion                                                                                     29

5.2       Conclusion                                                                                    31

References

LIST OF TABLES

Table 1: Total Viable Bacteria Count (cfu/ml)                                        24

Table 2: Range of Microbial Load of Street Vended Fruit Sample         25

Table 3 Incidence of bacteria Isolates on Fruit Sample                           26

Table 4: Colonial Morphology, Microscopy and Biochemical Characteristics of Bacteria                                                                        27

Table 5: Antimicrobial Sensitivity test for Gram positive Bacteria Isolate 28

CHAPTER ONE

1.0       INTRODUCTION

Fruits are essential parts of the diet of humans. Fruits and vegetables are rich in nutrients, micronutrients and vitamins which help the consumers with vitamin deficiencies, malnutrition and to get rid of the diseases related to these. Fruits contain good amount of vitamins mainly vitamin C and A, which are required for the treatment of most of the infections as a supplement along with medicines (KaliaI and Gupta, 2006). In common language usage fruit normally means the fleshy seed associated structures of a plant that are sweet or sour and edible in the raw state such as apples, oranges, grapes, strawberries, bananas, pawpaw and lemons (Schlegel, 2003). While according to oxford learner’s dictionary, "vegetables" can be any plant or a plant rose for some edible part of it, such as the leaves, roots, fruits or flowers. Examples are pumpkin, cucumber, cabbage, tomatoes, lettuce, spinach etc. Fruits and vegetables maybe be eaten pre-cut or sliced into pieces but remain in the fresh state and are stored or displayed for sale or for serving in retail outlets (fresh fruit and vegetable packs in supermarkets, cut fruits in buffet) assorted fruits offered by restaurants and condors (Kaplan and Campbell, 1982; Land, 1992; De Roever, 1998). Ready to eat fruits (street vended fruits) are fruits and vegetables prepaid by vendors for immediate consumption. According to Oranusi and Olorunfemi  (2011) ready to eat fruits and vegetables that can be bought directly from street hawkers or at local markets and eaten immediately without necessarily having to  cut, peep or rinse them before consumption as they have already been prepared by the vendors.

1.1       Nutritional Value of Ready to Eat Fruits and Vegetables

Fruits and vegetables are an extraordinary dietary source of nutrients, micronutrients, vitamins and fiber for humans and are thus vital for health and wellbeing. Well balanced diets, rich in fruits are especially valuable for their ability to prevent vitamin C and vitamin A deficiencies and are also reported to reduce the risk of several diseases (Kalia and Gupta, 2006).

1.2       Rate of Consumption of Ready to Eat Fruits and Vegetables

Over the years, there has been a significant increase in the consumption of sliced / ready to eat fruits and vegetables because they are easily accessible, convenient, nutritious and mostly cheaper than the whole fruits (Nwachukwu et al., 2008). Regular consumption of fruit us associated with reduced risks of cancer, cardiovascular disease (especially coronary heart disease stroke, Alzheimer disease, cataracts and some of the functional declines associated with aging (Liu, 2003)

1.3       Contamination of Fresh Fruits and Vegetables

Fruits and vegetables are exposed to microbial contaminants that are mainly due to high nutrition, neutral PH, micro nutrition and other favorable conditions required for the microbial growth. Fruits and vegetables harbor both human and plant microbes. Since they are widely exposed to the sewage contaminated soil, water and manures (Nguyen and Carlin, 1994; Carmo et al., 2004). These fruits can also get contaminated with pathogenic microbes during harvesting, transportation, storage, transport containers, retailer handling etc. Due to these wide range of exposure to different microbial flora of the fruits greatly vary (Ray and Bhunia, 2007; Ofor et al., 2009). Most microorganisms that are initially observed on whole fruit or vegetable surfaces are soil inhabitants which have members of a very large and diverse community of microbes. Transmission of microbes to these fruits and vegetables may also occur through vectors like soil particles, airborne spores and irrigation water. Microbes that harbor on the surface of fruits and vegetables may cause spoilage due to favorable conditions found for their growth.

1.4       Microbial Infection and Fresh Fruit and Vegetables

An increased number of microbial infections associated with consumption of fresh vegetables and fruits have been reported in recent years. Documented illnesses have been caused by bacteria, parasites, viruses and are transmitted through many types of fruits and vegetables (Beuchat, 1996; Nyenje et al., 2012).

1.5       Use of Antimicrobials in the Control of Pathogens

Excessive and misuse of antimicrobials to control pathogens in animals and crops led to antimicrobial (antibiotic) resistance and transfer to human through contaminated food. Some researchers show that antibiotic resistant bacteria may also be ingested with fruits and vegetables (Kilonzo et al., 2009)

1.6       Aims and Objective of this Research Work

A)        To ascertain the antimicrobial susceptibility or sensitivity pattern of microorganisms found or ready to eat fruits and vegetables within the surrounding of Michael Okpara, Federal University of Agriculture, Umudike, Abia State.

B)        To isolate and characterize the bacteria present in such foods around the university vicinity.

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