ANTIBIOGRAM OF ORGANISMS OF PUBLIC HEALTH IMPORTANCE FROM VEGETABLE WASH WATER

ABSTRACT

The antibiogram of organisms of public health importance from vegetable wash water was investigated. Twenty five (25) samples of vegetable wash water from five markets in Umuahia were analyzed. Four bacteria genera: Staphylococcus aureus, E.coli, Shigella spp, salmonella spp were isolated. Staphylococcus areus gave the highest percentage occurrence of 90% while Salmonella spp gave the least percentage occurrence of 30%. The Kirby Bauer disk diffusion method was used for the antibiotic susceptibility test. Staphylococcus aureus was highly susceptible to Ciprofloxacin, Rifampicin and Amoxil while others exhibited least susceptibility. Among the Gram negative organisms E.coli, Shigella spp and salmonella spp  were found to be highly susceptible to Ofloxacin, Gentamycin, Augumetin while other species of E.coli, Shigella and Salmonella gave the least susceptibility to Nalidixic acid, Penicillin, Amoxil, Streptomycin, Ciprofloxacin, Levofloxacin and Erythromycin.  The Multiple Antibiotic Resistance Index of the organisms ranged between 0.2-0.7. The result of this study showed that vegetable can be an avenue for transferring antibiotic resistant genes in community setting. Hence it is necessary to monitor and understand the critical points in the control of antibiotic resistance among the developing countries.

TABLE OF CONTENTS

Title page                                                                                                                                i

Certification                                                                                                                            ii

Declaration                                                                                                                             iii

Dedication                                                                                                                               iv

Acknowledgements                                                                                                                v

Table of contents                                                                                                                    vi

List of Tables                                                                                                                          vii

Abstract                                                                                                                                   viii

CHAPTER ONE

1.1       INTRODUCTION                                                                                                      1

1.2       Aims and Objectives                                                                                                  4

CHAPTER TWO

2.1       LITERATURE REVIEW                                                                                           5

2.2       Food-borne illness outbreaks linked to fresh produce                                               7

2.3       Sources of contamination                                                                                           10

2.4       Microorganisms Associated with Food Items                                                                        13

2.41     Bacillus species                                                                                                          13

2.4.2    Streptococcus species                                                                                                 14

2.4.3    Vibrio cholerae                                                                                                           15

2.4.4    Staphylococcus species                                                                                               15

2.4.5    Salmonella species                                                                                                     16

2.4.6    Enterobacter species                                                                                                  17

2.4.7    Pseudomonas species                                                                                                 17

2.4.8    Campylobacter species                                                                                               18

2.4.9    Shigella species                                                                                                          18

2.4.10  Escherichia coli                                                                                                        19

2.4.11  Aspergillus species                                                                                                     20

2.4.12  Fusarium species                                                                                                        20

2.5       Post-harvest Wash and Limitations                                                                            20

2.6       Wastewater Characteristics                                                                                        22

2.7   Wastewater treatment and recycling                                                                  23

CHAPTER THREE

MATERIALS AND METHODS                                                                                           27

3.1       Study Area and Sample Collection                                                                            27

3.2       Sterilization of Materials                                                                                            27

3.3       Preparation of culture media                                                                                      27

3.4       Isolation and enumeration of isolate                                                                          28

3.5       Purification of isolate and preservation or identification of isolates                                    28

3.6       Motility                                                                                                                       29

3.7       Biochemical Tests                                                                                                      29

3.8       Determination of Antimicrobial Susceptibility Pattern                                              31

CHAPTER FOUR

4.1       RESULTS                                                                                                                   32

CHAPTER FIVE

5.1       DISCUSSION                                                                                                             38

5.2       Conclusion                                                                                                                  39

5.3       Recommendations                                                                                                      39

References                                                                                                                              40

Appendix

LIST OF TABLES

Table          Title                                                                Pages

2.1                  General water quality parameters                                                                   25

4.1                   Characterization and identification of isolates from vegetable wash water 33

4.2                   Total aerobic count of vegetable wash water from different location        34

4.3                   Occurrence and percentage distribution of isolates from vegetable wash

water                                                                                                               35

4. 4                  Diameter  Zone of Inhibition of Antibiotics (in Mm) for gram Positive

Organism Isolated and MARI                                                                         33

4.5                  Diameter Zone of inhibition of Antibiotics for Gram Negative Organism

Isolated in (Mm) and MARI                                                                           34

LIST FIGURE

Figure           Title                                                                Pages

2.1          Water treatment process.                                        26

CHAPTER ONE

INTRODUCTION

Vegetables include the leafy, fruit, seed or root vegetables that are fit for human consumption. These parts are consumed whole or in part, raw or cooked as a supplement to other food crops. Nowadays, there has been an increase in fresh vegetables consumption due to health benefits in eating natural, healthy and functional foods. Consequently, consumers are seeking for vegetables products that maintain its natural nutrition and other aesthetic qualities such as flavor, texture and colour properties’ (Ahmed et al., 2014; Abadias et al., 2008). In the developing countries vegetables are comparatively cheap and easy to grow. Thus, most rural women rely on these vegetable as source of income. Despite the numerous benefits of vegetables, it could be a medium for the spread of bacterial, parasitic, viral pathogens and antimicrobial resistance mechanisms (Bannerman and Peacock, 2007). As a consequence, the production and distribution of fresh vegetables could be a huge challenge to the food industry.

Water is one of the principal agents for the spread of various enteric diseases (Saranraj et al., 2012). Some researchers have stated that water from contaminated sources can propagate harmful microorganisms that can affect the safety of such vegetables (Ahmed et al., 2014; Smetanska et al., 2013; Belguith et al., 2009). Since most of the populations that grow vegetables are from rural areas, majority of them cannot afford expensive technologies, such as use of chlorinated water to rinse their vegetables. Accordingly, they adopt locally- available and cheap technologies such as rinsing with ordinary water from rivers and shallow wells without recourse about the safety of the vegetables.

Food borne outbreaks associated with vegetables that are partially processed or consumed naturally have increased over the years (Khan et al., 2014; Beuchat, 2002). This rise in the food-borne outbreaks from vegetables is due to the favorable conditions that the vegetables provide consequently encouraging the growth and survival of many types of microorganisms. Some of the favorable conditions include nutrient rich internal tissues-comprising polysaccharides (starch), pectin, hemicelluloses and cellulose.

Enteropathogens are among the greatest concerns with food-borne outbreaks. In 2007, these organisms were implicated in food-borne outbreaks in UK that resulted in the recalling of bagged lettuce (Oyetibo et al., 2010). Such recalls damage the consumer’s confidence and hampers economically the income and corporate image of such food processing industries involved with the sale and distribution of such products. The enteropathogens have been implicated in cases of diarrhea and typhoid fever following consumption of contaminated vegetables (Clinical and Laboratory Standards Institute, 2011).

In most developing countries, street vending of fresh vegetables are on the increase and as such precautionary measures on the safety of the vegetables are not considered. Consequently, such vegetables could be a repository for various organisms that can severely affect the welfare of the consumers, shelf-life and nutritional worth of the vegetables. A study conducted in Ado-Odo Ota, Nigeria, selected vegetables popularly grown and consumed were assessed for epiphytic bacteria species associated with it and their antibiotics susceptibility patterns. The selected vegetables include: Corchorus olitorius (L.), Vernonia amydalina Delile, Abelmoschus esculentus and Celosia caracas. The selection of these vegetables were based on the huge culinary and medicinal benefits they offer the population when consumed raw or lightly cooked. For instance Corchorus olitorius when boiled or mashed as fresh leaves serves as rich sources of vitamins A, C and methionine’ (Cabral, 2010; Chigor et al., 2013). Also, Vernonia amydalina Delile is known for its antimalaria, antihelmenthic, antitumorigenic properties as well as antioxidant properties (Feroz et al., 2013).

In the farm-to-table production, important points of contamination of vegetables with disease-causing microorganisms include use of pond and river water to wash vegetable produce, handling of vegetables by workers infected with and carriers of the pathogen(s) and storage of vegetables in contaminated places. Considerable information is available on bacterial load in washing water as well as on vegetables and contamination of plant products by human pathogens is a known fact. Although it is not easy to establish exactly the level of disease risk associated with a given concentration of fecal coliforms or total aerobic plate count or yeast and mould counts, existence of such standards necessitate their observance to exist in international trade of food vegetables. In India, vegetables are washed mostly in readily available water bodies, such as rivers or ponds available in the vicinity of the production site. After the farmers, vegetable vendors are the important handlers of vegetables, selling different type of water source also influence the microbiological quality of produce, wastewater contaminate vegetables with pathogenic microorganisms (Solomon et al., 2002; Kumar, 2012).

It is now commonly accepted that fruit and vegetable consumption is a risk factor for infection with enteric pathogens (Heaton and Jones, 2008). Escherichia coli, Clostridium botulinum, Salmonella, Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus, grow on lettuce, cucumber, carrot, red cabbage, tomatoes and a variety of salad vegetables have been associated with food borne illness for decades (Borch and Arinder, 2002; Velusamy et al., 2010). Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica and Escherichia coli are known as common food borne pathogenic microorganisms (Kim et al., 2013). Most of the pathogens are not newcomers and food-borne outbreaks or infections are not a rare event (Harris et al., 2001; Velusamy et al., 2010). The symptoms of staphylococcal food poisoning had already been described in 1936, one of the earliest food poisoning caused by Bacillus cereus was reported in 1906 (Borch and Arinder, 2002). Cross-contamination is an important factor of food-borne illness. Fruits and vegetables become contaminated with pathogenic microorganisms while growing infields, orchards, vineyards or greenhouses, or during harvesting, post-harvest handling, processing, distribution and preparation.

Vegetables and fruits produce have potential to harbor pathogenic microorganisms, but Shigella spp, Salmonella, Escherichia coli, Campylobacter spp, Listeria monocytogenes, Yersinia enterocolitica, Bacillus cereus, Clostridium botulinum and parasites such as Giardia lamblia, Cyclosporacayetanensis and Cryptosporidium parvum are of greatest public health concern (Beuchat, 2002).

Recently, efforts have been devoted to prolonging the shelf life of vegetables, however, the determination of the antibiogram patterns of the bacteria species associated with these vegetables have been scarcely reported.

1.2       Aims and Objectives

The objective of this study is;

i.               To isolate and identify organisms of public health importance from vegetable wash water.

ii.              To determine the microbial load of the vegetable wash water.

iii.            To determine the antibiogram of organisms of public health importance isolated.

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