ISOLATION AND ANTIBIOTIC SENSITIVITY TEST OF PATHOGENIC BACTERIA FROM READY-TO-EAT FOOD

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ABSTRACT

Today the issue of food safety is a global problem that gets main concern in setting public health policy. The eruption of diseases caused by food contamination occurs in places where sanitation and hygiene conditions are generally poor. This study was conducted to isolate and determine the antibiogram of bacteria pathogens in ready-to-eat foods sold in MOUAU campus. Four ready-to-eat food samples were collected from different vendors within the campus and analyzed immediately through serial dilution, inoculation, incubation, subculture, microscopy and biochemical to confirm the isolated organisms. Antibiotics sensitivity test was also carried out to determine antibiotic sensitivity on the isolated organisms. A total of four pathogenic bacteria were isolated: Staphylococcus aureus (40%), Salmonella (17.5%), Escherichia coli (30%) and Shigella (12.5%). Eight different antibiotic were used to carry out antibiogram of the isolates as the inhibition zone was measured in millimeter: Oflovacin (OFX), Amoxil (AMX), Gentamycin (CN), Augmentin (AUG), Ciprofloxacin (CPX), Chloramphenicol (CH), Ampiclox (APX) and Streptomycin (S). (AUG) and (CPX) had high effect on Salmonella and Escherichia coli and was less on Staphylococcus aureus while (OFX) and (S) were very effective on Staphylococcus aureus and less effective on Escherichia coli and Salmonella. This shows the poor hygiene level of the food vendors as majority of the food vendors lacks western education. These vendors have to receive education and training on food hygiene to improve the safety of foods in MOUAU and thereby heighten the safety of consumers.





TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                           iii

Dedication                                                                                                                              iv

Acknowledgments                                                                                                                  v

Table of contents                                                                                                                    vi

List of Tables                                                                                                                          ix

Abstract                                                                                                                                  x

 

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1       Aim and objective                                                                                                      3

 

CHAPTER TWO

2.0       Literature review                                                                                                        4

2.1       Food                                                                                                                            4

2.2       Classification of foods                                                                                                4

2.2.1    Adulterated food                                                                                                         4

2.2.2    Diet food                                                                                                                     4

2.2.3    Finger food                                                                                                                 4

2.2.4    Functional food                                                                                                           5

2.2.5    Health food                                                                                                                 5

2.2.6    Natural food                                                                                                                5

2.3       Factors that influence microbiological activity in ready to eat food                                    6

2.4       Ready to eat food                                                                                                        9

2.4.1    Source of ready-to-eat food contamination                                                                9

2.5       Food-borne illness                                                                                                      10

2.5.1    Causes of food-borne disease                                                                                     10

2.5.1.1 Bacteria                                                                                                                       11

2.5.1.2 Mycotoxins                                                                                                                 11

2.5.1.3 Natural toxins                                                                                                             11

2.5.1.4 Viruses                                                                                                                        12

2.6       Pathogenic bacteria                                                                                                    12

2.6.1    Common pathogenic bacteria associated with ready to eat food                                        14

2.6.1.1 Staphylococcus aureus                                                                                               14

2.6.1.2 Salmonella                                                                                                                  14

2.6.1.3 Escherichia coli                                                                                                          14

2.6.1.4 Shigella                                                                                                                       14

2.7       Ways of preventing pathogenic bacteria contamination of ready-to-eat food       15

 

CHAPTER THREE

3.0       Materials and method                                                                                                 16

3.1       Sample collection                                                                                                       16

3.2       Sterilization of glasswares and media                                                                        16

3.3       Microbiological analysis                                                                                            16

3.4       Procedure                                                                                                                    16

3.4.1    Serial dilution                                                                                                             16

3.4.2    Inoculation                                                                                                                  17

3.4.3    Incubation                                                                                                                   17

3.4.4    Colony counting                                                                                                         17

3.4.5    Isolation of pure culture                                                                                             18

3.5       Identification of bacteria isolates                                                                               18

3.5.1    Macroscopic examination                                                                                          18

3.5.2    Microscopic examination                                                                                           18

3.5.3    Gram staining                                                                                                             18

3.6       Biochemical test                                                                                                         19

3.6.1    Indole test                                                                                                                   19

3.6.2    Catalase test                                                                                                                19

3.6.3    Coagulase test                                                                                                             19

3.6.4    Oxidase test                                                                                                                20

3.7       Antibiogram susceptibility test                                                                                   20

 

CHAPTER FOUR

4.0       Result                                                                                                                          21

 

CHAPTER FIVE

5.0       Discussion, Conclusion, Recommendation                                                                25

5.1       Discussion                                                                                                                   25

5.2       Conclusion                                                                                                                  26

5.3       Recommendation                                                                                                       26

References                                                                                                                  27

 

 

 




 

LIST OF TABLES


4.1       Shows the percentage occurrence of the isolates                                                       22

4.2       Shows the identification of the isolates                                                                      23

4.3       Shows the inhibition zones of the antibiotics on the isolates                                               24

 

 


 




 

CHAPTER ONE


1.0     INTRODUCTION

 Ready-to-eat food (RTE) is defined as food that is ordinarily consumed in the same state as that which it is sold. Ready to eat food like jollof rice, meatpie, eggroll, sandwish, snacks moi-moi are very essential to consumers as it reduces the time at which food is to be prepared (Wikipedia, 2019). Ready to eat foods usually include a number of ingredients which may or may not be cooked. Some ready to eat foods also are regarded as potentially hazardous, such foods can support the growth of pathogenic (food poisoning) bacteria and must be kept at certain temperatures to minimize the growth of any pathogen that may be present in the food or to prevent the formation of toxins in the food (New South Wales, 2014). It was reported that 42% of food-borne diseases outbreaks which took place in America have been caused by food handlers (Sadiq, 2008).

 

In developing countries such as Nigeria, there are serious concerns about sanitation of ready-to-eat foods, particularly as potable water is soldem available at preparation venues and fast food stands and also most food handlers lack basic knowledge of proper personal and environmental hygiene (Bukar et al., 2010). Although it’s extremely difficult to pinpoint the precise beginning of human awareness of the presence and role of micro-organisms in foods, the available evidence indicates that this knowledge preceded the establishment of bacteriology or microbiology as a science. The microbial spoilage of foods may be viewed simply as an attempt by the food biota to carry out what appears to be their primary role in nature. Outbreaks of food borne diseases are caused by foods that are contaminated intrinsically or that become contaminated during harvesting. The consumption of food contaminated by micro-organisms will result in food- borne illnesses; these are usually either infectious or toxic in nature caused by agents that enter the body through ingestion of food (WHO, 2013). According to the U.S. department of Health and Human Services (USDHHS) website, food -borne illnesses are diseases that results from eating contaminated food (USDHHS, 2013).

 

Food- borne illnesses have continued to form a significant part of the morbidity and mortality of Nigerians, and have on the increase in recent times. In Nigeria and many developing countries, there are inadequate diagnostic facilities leading inadequate investigation of outbreaks and the subsequent gross under-reporting of food-borne illnesses. In Nigeria, a number of foods have been reported to have high incidence of bacteria (Adesetan et al., 2013; Bello et al., 2013). According to surveys carried out in 2006 and 2007 by the department of Public Health of the Federal Ministry of Health, Nigeria, there were more than two million recorded cases of food borne diseases in the country with the number of death put at over five hundred. Some of these cases could be linked to consumption of contaminated street vended foods (Agu, 2011). A number of observational studies have shown that ready to eat foods are sometimes held at improper temperatures excessively handled by food vendors and sold at very dirty surroundings. Lacking personal hygiene among food handlers is one of the most commonly reported practices contributing to food-borne illness and poor hand and surface hygiene is also a significant contributory factor (WHO, 2013).

 

The hands of food handler can be pivotal as vector in the spread of food-borne diseases due to poor personal hygiene or cross-contamination. Hand washing, a simple and effective way to cut down on cross-contamination is too often forgotten. Microbial contaminants such as bacteria constitute the major cause of severity ranging from mild indisposition to chronic or life threatening illness or both (Ibrahim et al., 2013). In developing countries, such contaminants are responsible for food-borne disease such as cholera, Escherichia coli gastroenteritis, salmonelosis, shigellosis, typhoid fever, etc. Safe food is basic human rights despite many foods are frequently contaminated with naturally occurring pathogenic micro-organisms, such pathogens cannot be detected organoleptically, but can cause diseases of varying severity including death especially in the way they are conserved during exposition for sales provides condition for those micro-organisms to grow and reach considerable levels of contamination. Thus, food safety issues are of major important issues to the World Health Organization.

 

AIM AND OBJECTIVE

The aim and objective of this study are:

1.     To isolate pathogenic bacteria from ready-to-eat food sold in Michael Okpara University of Agriculture Umudike, Abia state.

2.     To determine the antibiogram of the bacteria isolated.

 


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