ISOLATION AND IDENTIFICATION OF BACTERIA ASSOCIATED WITH

WATERMELON AND CARROT SOLD AT WUDIL MARKET, WUDIL L.G.A. KANO

STATE
CONDUCTED AT ALIKO DANGOTE UNIVERSITY OF SCIENCE AND
TECHNOLOGY, WUDIL

BY

SAFIYANU IBRAHIM
UG20/MICR/2013

FACULTY OF SCIENCES
DEPARTMENT OF MICROBIOLOGY
ALIKO DANGOTE UNIVERSITY OF SCIENCES AND TECHNOLOGY WUDIL
SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENT
OF THE AWARD OF BSC.(HONS) IN MICROBIOLOGY

SUPERVISED BY
Prof. MUHAMMAD YUSHA'U
.

i
 20th AUGUST, 2024
CERTIFICATION

This is to certify that this work on isolation and identification of bacteria associated with

watermelon and carrot was carried out by SAFIYANU IBRAHIM with registration number

UG20/MICR/2013 and is the bonafide work of him submitted to microbiology department of

Aliko Dangote University of Science and Technology Wudil.

The research work was carried out by him under the supervision and guidance of Prof.

MUHAMMAD YUSHA'U

Prop MUHAMMAD YUSHAU ________________________

Supervisor Signature Date
.

Mal. TASIU MUHAMMAD __________________________

Coordinator Signature Date

Dr. ADAMU ABDULLAHI SHEHU _________________________

Head Of Department Date

ii
 DEDICATION

As well as everything that I do. I would be honor to dedicate this compilation to my parents. The

two person that gave tools and values necessary to be where I am standing today. My parents

support me on every step i make and decision I take, but is necessary to understand that they let

me take my decisions alone in order for me to learn my personal mistake and as my father says

to "learn and grow from each setback". I will never finish to thank my parents for all the

opportunities that they have offer and gave me, for all the teachings that they have told me and

for every advise that comes out from their mouth.

I'm so grateful with them for trusting me to that I would do a good job in the university, and

letting me come to achieve a higher education.

iii
 ACKNOWLEDGEMENT

Alhamdulillahi Rabbil Alamin, I give thanks to Almighty Allah, Al-hayyu Al-qayyum, zuljalalu

wal ikram for his protection, guidance and for giving me the ability to reach these [Link] is not

just a duty but also an honor to acknowledge the attention and support I received during this

review on food borne illnesses. My deepest and warmest appreciation goes to the Prof.

MUHAMMAD YUSHA'U for his r support, motivation, and advises given to me, may Allah

rewards him abundantly.

Lastly, without his help along the way, I'm not sure if I could have made it here today, so thanks

go out as well to everyone else that contributed at some point or another during my journey on

completing this remarkable undertaking report .

iv
 ABSTRACT

Isolation and identification of bacteria was carried out on watermelon and carrot sold in Wudil

Market, Wudil L.G.A Kano State . Eight (8) samples of watermelon and carrots were collected

randomly from different stationery vendors and analyzed immediately through serial dilution,

inoculation, incubation, subculture, microscopy and biochemicals tests to confirm the isolated

organisms using standard [Link] bacterial and total coliform count was carried out on

all samples. The bacterial load ranged from 0.90x 10–¹, 1.10 x 10–¹,1.00 x 10–¹, 1.30x 10–¹ (for

watermelon-1 to 4) and 1.00 x 10–¹, 1.00 x 10–¹, 1.11 x 10–¹ , 1.14 x 10–¹ (Carrot- 1 to 4) and

Total coliform count ranged from <3 to <4 MPN/g. The analysis has shown that watermelon and

carrot sold in Wudil Market contain considerable numbers of pathogenic bacteria.

v
 TABLE OF CONTENT

Certification.....................................................................................................................................ii

Dedication......................................................................................................................................iii

Acknowledgement..........................................................................................................................iv

Abstract............................................................................................................................................v

Table of Content.............................................................................................................................vi

CHAPTER ONE..............................................................................................................................1

1.0 Introduction................................................................................................................................1

1.1 Background Of The Study.........................................................................................................1

1.3 Justification................................................................................................................................4

1.4 Aim and objectives....................................................................................................................4

CHAPTER TWO.............................................................................................................................5

2.0 Literature review........................................................................................................................5

2.1 Food borne illnesses..................................................................................................................9

2.2 Features of common food–borne pathogens............................................................................11

2.3 Lessons Learnt And Future Challenges...................................................................................14

2.4 Pre-disposing factors to food borne illnesses..........................................................................16

2.5 Causes Of Food Borne Illnesses..............................................................................................18

2.6 Who Gets Food Borne Illnesses..............................................................................................20

2.7 Symptoms Of Food Borne Illnesses........................................................................................20

2.8 The Complications Of Food Borne Illnesses...........................................................................21

2.8.1 Treatment Of Food Borne Illnesses......................................................................................22

vi
2.8.2 Factors That Contribute To Food Borne Illness...................................................................24

2.8.3 Prevention And Control Of Food Borne Illnesses................................................................25

2.9 “Some Golden Rules” Of Who For Safe Food Preparation (10).............................................27

CHAPTER THREE.......................................................................................................................28

3.0 Material and Methods..............................................................................................................28

3.1 Study Area...............................................................................................................................28

3.2 Collection Of Samples.............................................................................................................28

3.3 Sterilization Of Materials........................................................................................................28

3.4 Preparation Of Culture Media.................................................................................................28

3.7 Isolation And Enumeration Of Bacteria..................................................................................29

3.7.1 Sub- Culture Technique........................................................................................................30

3.8 Enumeration Of Coliform Bacteria.........................................................................................30

3.9.1 Data Analysis........................................................................................................................33

CHAPTER FOUR.........................................................................................................................34

4.0 Result.......................................................................................................................................34

4.1 Total Bacterial Count :.............................................................................................................34

4.2 Total Coliforms Count.............................................................................................................35

4.3 Gram Stain Reaction:...............................................................................................................36

4.5 Biochemical reaction of the Bacterial Isolates.:......................................................................37

4.6 Discussion................................................................................................................................38

CHAPTER FIVE...........................................................................................................................41

5.0 Summary, Conclusions And Recommendations.....................................................................41

5.1 Summary..................................................................................................................................41

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5.2 Conclusions..............................................................................................................................41

5.3 Recommendations............................................................................................................................42

REFERENCES..............................................................................................................................43

viii
 CHAPTER ONE

1.0 INTRODUCTION

1.1 Background Of The Study

The consumption of fruits and vegetables have notable beneficial effect as it contains essential

nutrients that support good health (Liu 2013), which include fiber, vitamins and essential

minerals amongst others. Consumers of fresh fruits and vegetables are usually in a state of good

health, hence the several health benefits being enjoyed (Adebolu and Ifesan, 2019).There is a

drastic reduction of incidences of acute and chronic diseases as a result of the intake of fruits and

vegetables (Boeing et al., 2012).

There is considerable information on the need to consume fruits and vegetables on a daily basis

as essential part of our diet, and it is also known that there could be development of poor health

and increased incidences of disease due to insufficient consumption (Aune et al., 2017).

However, the intake of raw vegetables without proper washing is a great concern because it has

been proved to harbor microbes (WHO, 2015). It has also been documented by Temgoua et al.,

(2015) and Adams and Moss (2018) that contamination could arise from poor or lack of basic

standard systems deployed during post harvest transportation, handling and packaging.

Fruits and vegetables could be the sources of contamination of food preparation areas (Altieri

and Nicholls, 2017). Ready to-eat fruits and vegetables may be sliced, peeled, shredded and

washed/unwashed (Francis et al., 2012). The destruction of surface cells during processing can

cause an exposure of the produce for the entry of microorganism which utilizes the readily

available nutrients compared to intact produce. In addition, high water activity of many fruits and

approximately neutral pH of vegetables encourages the rapid growth of microbes (Qadri et al.,

2015). Members of the Enterobacteriaceae is often associated with contamination of fruits and

1
vegetables due to activities and processes involved from cultivation and post harvest operations.

Varieties of fruits and vegetables have been implicated as a source of Salmonella infection; most

commonly are tomatoes, lettuce and watermelon. Salmonella sp. Has been found on fresh

produce such as lettuce, cauliflower, spinach, mushrooms and mustard cress (Mritunjay and

Kumar, 2015).

Food safety is a growing concern for the consumers and professionals in food and food service

industry (Adesiyun, 1995). Food safety is defined as the conditions and measures that are

necessary during production, processing, storage, distributions and preparation of food to ensure

that it is safe, sound wholesome and fit for human consumption (Adesiyun, 1985). Food hygiene

is essentially aimed at producing food which is safe for human consumption and of good keeping

quality (Scheule, 2001).

Microbial contaminants such as bacteria constitute the major cause of severity ranging from mild

indisposition chronic or life threatening illness or both (Ibrahim, 2013). In developing countries,

such contaminants are responsible for food-borne disease such as cholera, Escherichia coli

gastroenteritis, salmonelosis, shigellosis, typhoid fever, [Link] study was carried out to appraise

the bacteriological quality and hygiene level of some food outlets within the Wudil Market.

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 (WHO, 2015). The study is aim at analyzing bacteria on

watermelon and carrot sold at Wudil Market, Wudil L.G.A. Kano State, Nigeria, as very little or

2
no information is available.

Generally, most of the street vendors that sales ready fruits and vegetables are not monitored by

any food protection agency which means the risk of consuming fresh produce contaminated with

infectious agent such as Escherichia, Salmonella and other parasite is high (Orji et al., 2017).

The ability of some certain bacteria to produce toxin causing food poisoning has also implicated

Staphylococcus as one of the prevalent agent responsible for infection and can be transmitted

from person to person (foster and McDevitt, 2013). The entirely study was aimed at assessing

bacterial contamination in ready to eat fruits and vegetables sold at Wudil Market, Wudil L.G.A.

Kano State.

1.2 Problem Statement

Fruits and vegetables are often consumed without any further processing, which increases the

risk of foodborne illnesses. The presence of pathogenic bacteria on these produce items can lead

to outbreaks of foodborne diseases, posing a significant public health risk.

Current methods for isolating and identifying bacteria may not comprehensively address the

broader spectrum of bacterial communities and potential pathogenic infiltration.

Therefore, there is a need for effective isolation and identification techniques to ensure food

safety and mitigate microbial [Link] illnesses remain a frequent and

significant hazard to public health worldwide

Fruits have been connected to several outbreaks of foodborne illness globally, including in

Nigeria. Understanding the bacterial communities on fresh produce is crucial for assessing the

risk of foodborne illness and spoilage associated with fruits and vegetables.

This knowledge can inform targeted interventions to improve food safety and protect public

health.

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1.3 Justification

Bacteria produce toxins, which pose health risks to humans, causing diseases and various health

issues (Bennett & Klich, 2009; Binder et al., 2012). Bacterial contamination also leads to

economic losses for farmers by reducing feed efficiency and productivity, and resulting in the

rejection of contaminated products in the market (Ayo et al., 2018). Additionally, there is a lack

of awareness among farmers about the risks of bacterial contamination, leading to poor feed

management practices (Fagbohun & Lawal, 2012). The absence of stringent regulations further

exacerbates the [Link] research will help mitigate health risks, reduce economic losses,

improve awareness, inform regulatory policies, and fill critical knowledge gaps, ultimately

enhancing the safety and quality of fruits and vegetables sold in Wudil Market, Wudil L.G.A,

Kano State.

1.4 AIM AND OBJECTIVES

Aim

The aim of this study was to isolate and identify bacteria associated with watermelon and carrot

sold in Wudil market.

Objectives

1- To determine the bacterial load in the watermelon and carrot.

2- To determine coliform bacteria of public health important

3- To identify bacteria in the samples (watermelon and carrot)

4
 CHAPTER TWO

2.0 LITERATURE REVIEW

Foodborne illnesses remain a significant public health concern worldwide, with contaminated

fruits and vegetables from vendors being a notable source of bacterial pathogens. Before 1960

the major causes of gastrointestinal disease were recognised as Salmonella spp., Shigella spp.,

Clostridium botulinum and Staphylococcus aureus. During the 1960s Clostridium perfringens,

and B. cereus were added and then in the 1970s, rotavirus and norovirus. In the 1980s and 1990s

there was a flurry of additions including Campylobacter, Yersinia, Listeria monocytogenes, new

strains of Escherichia coli such as O157:H7, Cryptosporidia and Cyclospora. It seems highly

probable that new food-borne pathogens will be discovered in the 21st century. Many of these

will be zoonotic in origin given that such pathogens are twice as likely to cause new and

emerging diseases than non-zoonotic agents. In addition, already known pathogens can evolve

thereby adding to the public health risks. For example zoonotic food-borne bacteria are

increasingly becoming resistant to antimicrobials. In 2005 it was reported that 1.8 million people

died from diarrhoeal diseases ([Link]/mediacentre/factsheets/fs237/en/), largely

attributable to contaminated food and drinking water. This is not just an underdeveloped world

problem. About 76 million cases of food-borne diseases, resulting in 325,000 hospitalizations

and 5000 deaths, are estimated to occur each year in the United States of America (USA) alone

(Mead et al., 1999). There are over 200 known microbial, chemical or physical agents that can

cause illness when ingested (Acheson, 1999). Over the last 20 years, at least in the industrialised

world, food-borne diseases caused by bacteria have significantly moved up the political agenda

and generated, on occasions, substantial media attention. In the face of such public concern,

public health efforts have been directed mainly towards the well-recognised food borne diseases

5
and pathogens in the food chain. In contrast, resources for emerging foodborne diseases have

tended to be allocated in an incident driven manner, i.e. in response to the extent of perceived

emerging health threat (for example BSE). In an attempt to reduce disease burden, the

monitoring of food-borne diseases and pathogens in the food chain has been implemented and a

farm-to-fork approach has been adopted encouraging all sectors of the food production chain to

improve hygiene and actively incorporate structured approaches to food safety, such as HACCP

principles.

E. coli O157 was first recorded in 1982 in outbreaks of severe bloody diarrhoea in North

America. Such outbreaks increased dramatically and became widespread in the following years.

It has been estimated that in the United States, E. coli O157:H7 causes 73,000 illnesses (Mead et

al., 1999) and 250 deaths annually. The first recognized community outbreak of O157:H7 in

Europe occurred in the United Kingdom in the summer of 1985 and further outbreaks and

sporadic cases have been reported throughout Europe ever since. In England and Wales, 33% of

outbreaks are food-borne (Gillespiel et al., 2005). In the early outbreaks the sources were most

often found to be contaminated beef meat, often minced, and the organism is widespread in the

guts of asymptomatic cattle. As a reflection of the seriousness of such outbreaks, extensive

legislation, new food handling practices and food-producer education have been introduced and

implemented over the last 20 years.

Food borne illness caused by microbial contamination of foods in an important international

public health problem and is known to be a major cause of diarrhoea diseased especially in

developing countries (Mensah, 2007). In these developing countries a major source of vended

foods or ready-to-eat foods are prepared and sold at public places such as markets place, schools,

and canteens and along the streets. The vended food is relatively cheaper and at easily accessible

6
places. Furthermore, it offers the traditional meals and preparations of a number of them are

quite laborious and time consuming (Amoah, 2012; Chakra Varky and Canet, 2012).

However, a number of observational studies have revealed that these items are occasionally

maintained at incorrect temperatures, handled excessively by food sellers, and served in filthy

environments (WHO, 2011, 2013; Ghosh et al., 2017). Furthermore, the vendors practice poor

personal cleanliness, and there have been numerous reports of food sellers being carriers and so

serving as a possible source of enteric fever transmission. Most of the sellers have either no

formal education or only a few years of schooling and hence lack understanding of basic food

handling and their role in pathogen transmission (Mensah et al., 2009). At the same time, the

majority of individuals who consume these foods are more concerned with their convenience

than with the bacteriological quality and sanitation of the food. The bacteriological quality of

food reflects the quantity of bacterial pollutants present; a high degree of contamination suggests

poor quality and an increased risk of illness transmission. Concerns have been expressed by the

Food and Agricultural Organization (FAO) and others that these foods could be a source of food

poisoning outbreaks (Chakravarty and Canet, 2012).

The 2015 WHO report on the estimates of the global burden of foodborne diseases highlighted

that more than 600 million cases of foodborne illnesses and 420,000 deaths could occur in a year,

with naturally occurring toxins and environmental pollutants being of most concern for health

In Nigeria, consumption of vended food has seen a phenomenal growth over the years as rapid

population growth, urbanization, unemployment, and poverty; occupational pressures, and

lifestyle changes have created a poll of mobile and transient population who rely almost entirely

on these relatively low cost foods for their nutrition (Martin, 2016).

7
Although epidemiological data on the incidence of food borne infections is insufficient, and

outbreaks are frequently not investigated, recurring episodes of food borne illnesses with

symptoms of gastro intestinal distress such as diarrhoea, vomiting, abdominal cramping, and

nausea have remained a major cause of mortality and morbidity in Nigeria (Nweze, 2010).

Food-borne sickness can be caused by chemicals, heavy metals, parasites, fungus, viruses, and

bacteria. Bacteria-related food poisoning is the most prevalent, yet only around 20 of the culprits

are known. Staphylococcus aureus, Salmonella, Clostridium perfringes, Clostridium botulinum,

Campylobacter, Bacillus cereus, and Entropathogenic Escherichia coli cause more than 90% of

food poisoning incidents each year. These bacteria can be found in a variety of raw foods.

Normally, a high number of food-poisoning bacteria must be present to cause disease; therefore,

illness can be avoided by reducing the amount of bacteria present, preventing the small number

from multiplying, killing the bacteria through correct cooking, and avoiding recontamination (De

Boer and Beuner, 2011).

Poor personal hygiene, inappropriate cleaning of storage and preparation rooms, unclean utensils

all contribute to raw and cooked food contamination. Bacteria can thrive when raw and cooked

foods are improperly handled. Most bacteria grow at temperatures around 37oC; foods, including

raw and cooked, should not be stored in this danger zone for any longer than absolutely

necessary. Analyzing foods for the presence of harmful and spoilage microorganisms is a

common practice for maintaining food safety and quality (De Boer and Beuner, 2011). If bacteria

can survive and thrive on food that is sold and consumed by people, the danger of food- borne

illness increases in society. The presence of microbes on food can be significant since the vital

nutrients taken by some organisms drive growth, but others are known to be dangerous to

humans as long as their growth conditions are favourable (De Boer and Beuner, 2011). Bacteria

8
can spread from equipment to food if the equipment that has come into contact with the food has

not been thoroughly cleaned and sanitized before being used to prepare another item (James,

2015). Food consumption has a direct impact on health; it is the responsibility of food

manufacturers and handlers to keep food safe from harmful microbes, especially when such

items are to be consumed without further processing (Munide and Kuria, 2015). When a food

containing hazardous bacteria is consumed, it takes some time before signs of food-borne illness

appear. The amount of time varies depending on the bacterium, the quantity consumed, and the

individual's physical state. Many different types of pathogenic microbes cause the same

symptoms (FDA, 2014).

2.1 Food Borne Illnesses

Foodborne illnesses are infections or irritations of the gastrointestinal (GI) tract caused by food

or beverages that contain harmful bacteria, parasites, viruses, or chemicals. The GI tract is a

series of hollow organs joined in a long, twisting tube from the mouth to the anus. Common

symptoms of foodborne illnesses include vomiting, diarrhea, abdominal pain, fever, and chills.

Most foodborne illnesses are acute, meaning they happen suddenly and last a short time, and

most people recover on their own without treatment. Rarely, foodborne illnesses may lead to

more serious complications. Each year, an estimated 48 million people in the United States

experience a foodborne illness. Foodborne illnesses cause about 3,000 deaths in the United

States annually. The global incidence of food borne disease is difficult to measure, although it

has been claimed that 2.1 million people died from diarrhoea infections in 2010 alone (WHO,

2011). Unsafe food causes a variety of acute and chronic ailments, ranging from diarrhoea to

cancer (WHO, 2011). According to the WHO, food borne and waterborne diarrhoeal illnesses

kill approximately 2.2 million people each year, with 1.9 million of them being children (WHO,

9
2011). In many regions of the world, the possibility of serious food poisoning outbreaks

associated to vended foods remains a concern, with microbial contamination being one of the

most serious issues (FAO, 2018). Food-borne infections are regarded as a major health risk

connected with vended foods, with the risk being mostly depending on the type of food and the

method of preparation and storage (WHO 2015). Every year, outbreaks of food- borne diseases

are reported in Kenya (MOH, 2013). Pathogenicity and virulence of an organism are controlled

by virulence coding genes found in pathogenicity islands in the genome (Hacker and Kaper,

2010). Staphylococcus aureus is the most common pathogen that has caused several outbreaks

(Veras et al., 2018). Staphylococcus aureus is a gram-positive bacteria that is positive for both

catalase and coagulase (Veras et al., 2018). Food contamination with enterotoxigenic bacterial

Staphylococcal enterotoxins (SEs) intoxication is caused by Staphylococcus aureus, which

results in symptoms such as vomiting and diarrhea. The following staphylococcal enterotoxins

have been identified: SEA, SEB, SEC, SED, and SEE (Robbins et al., 2017), as well as SEG,

SEH, SEI, SEJ, SEK, SEL, SEM, SEN, SEO, SEP, SEQ, and SEU, which were identified more

recently (Letertre et al ., 2013; Yarwood et al., 2012). The most prevalent SE related with food-

borne outbreaks is SEA, which is followed by SED. However, the kind of SE is unimportant

because SEs are structurally and functionally extremely similar (Balaban and Rasooly, 2010).

Shiga toxin-producing Escherichia coli are a group of bacteria strains capable of causing major

human disease, the infection is primarily spread through food (Richard, 2009).. The

enterohaemorrhagic E. coli subgroup comprises the relatively important serotype O157:H7, as

well as over 100 other non-O157 strains. Infection is spread usually through food and, less

frequently, through direct touch or water. Shiga toxins are produced by a variety of species,

including Shigella dysenteriae type I and Shigella toxin-producing Escherichia coli. These toxins

10
have a cytotoxic effect on intestinal epithelial cells, which is likely what causes the distinctive

bloody diarrhea (Richard, 2009). Identification of E. coli O157:H7 strains in the laboratory is

simple using specialized media, but identification of non-O15 Shigella toxin-producing

Escherichia coli strains requires detection of the Shigella toxin gene via polymerase chain

reaction or DNA probe-for virulence genes stx1, stx2, and eae (Richard, 2009). The

Enterococcus genus replaced faecal coliforms as the new federal guideline for water quality and

public beaches in Hawaii, USA, in 2004. It has a stronger relationship than faecal coliforms with

several of the human pathogens commonly detected in municipal sewage (Jin et al., 2010).

Enterococci, on the other hand, do not multiply in water, particularly in low organic matter

environments. They are fewer in number than Escherichia coli (James et al., 2015).

2.2 FEATURES OF COMMON FOOD –BORNE PATHOGENS

Salmonella Species

Salmonella: - is a generic term for a group of roughly 2,000 biochemically related serotypes that

cause food-borne disease. The disease is substantially underreported since it is typically self-

limiting gastroenteritis that might be mistaken as intestinal influenza by either the patient or the

clinician. As a result, estimates of illness incidence are based on assumptions obtained from

epidemiological evidence. Salmonellosis is clearly still a major cause of food-borne disease over

the world (Mead et al., 2009). Salmonella causes two clinical manifestations: enteric fever (a

severe, lifethreatening infection) and the more frequent food-borne illness syndrome. In both

circumstances, they are transmitted orally (Mead et al., 2009). Enteric fever, sometimes known

as typhoid fever, is caused primarily by one species, Salmonella typhi. However, other

Salmonellae, such as Salmonella paratyphi, have the ability to cause this disease. The sickness is

frequently related with international travel and affects an estimated 800 persons each year (Mead

11
et al., 2009). Although the virus enters the body largely through the mouth, symptoms of enteric

fever are not usually evoked through the gastrointestinal system. However, in the first day or two

of typhoid fever, a brief episode of vomiting and diarrhoea may occur. The onset times of

typhoid and paratyphoid enteric fevers differ significantly. The onset time for typhoid fever is

typically 8 – 15 days, seldom as short as five days, but sometimes as long as 30 – 35 days;

however the onset time for paratyphoid fever is typically shorter, and may be so quick as to

suggest ordinary food poisoning (Parker, 2014). Salmonella are killed during the cooking

process. Contamination of cooked foods happens as a result of interaction with utensils that have

not been thoroughly cleaned following usage with raw goods. If salmonella is present in raw or

cooked meals, its growth can be inhibited by keeping the temperature below 4oC. Salmonella are

transferred by the faeces matter of people or animals and are mainly transmitted to humans

through cross-contamination of foods that have been contaminated with faecal matter. To

produce sickness, as little as 15 to 20 cells are required, depending on the host's age, condition,

and strain of bacteria . Each year, around 40,000 cases of Salmonellosis are documented in the

United States (FDA, 2014).

Staphylococcus aureus

Staphylococcus aureus is commonly found in man's respiratory tract, skin, and superficial

wounds. Staphylococcus aureus can create a toxin that causes disease when allowed to develop

in foods. Although heating kills the bacteria, the toxin produced by the bacteria is heat stable and

may not be destroyed. Staphylococcal food poisoning is particularly common in dishes that

require extensive preparation. These foods are sometimes left at room temperature for extended

periods of time, allowing bacteria to proliferate and create toxin. Personal cleanliness when

12
handling foods will keep Staphylococcus aureus at bay, and chilling of raw and cooked meals

will stop the growth of these bacteria if any is present (Wagner, 2011)

Shigella

Shigellosis, also known as bacillary dysentery, is caused by bacteria of the genus Shigella, which

includes Shigella dysenteriae, Shigella flexneri, Shigella boydii, and Shigella sonnei (Bryan,

2009). Shigella's natural habitat is the human and other primates' intestinal tracts. The faecal-oral

route appears to be the primary method of transmission (Feldman and Riley, 2015). Shigella is

most commonly related with poultry, raw vegetables, dairy products, and poultry. These foods

are typically contaminated via the Faecal-Oral route, which is most commonly caused by

faecally contaminated water and unsanitary food handling by food handlers (Todar, 2016).

Disease can be caused by as little as 10 cells, depending on the host's age and health. Shigella,

like Escherichia coli, is present in the diarrhoea stool of an infected person and can be

transmitted during infection as well as one to two weeks after symptoms subside. Most infections

occur as a result of the bacterium passing from one person's stools or soiled fingers to the mouth

or finger of another person. In underdeveloped nations, Shigella dysenteriae causes lethal

epidemics (CDC – DBM D, 2014). Shigella is spread by the faeces of humans or animals and is

mainly transferred to humans by consuming foods contaminated with faeces due to cross

contamination. To produce food-borne illnesses, as little as 15 – 20 cells are required, depending

on the host's age and condition, as well as the type of [Link] borne shigellosis is

distinguished by a high attack incidence, a common source epidemiology, and short incubation

periods ranging from 12 to 50 hours (FDA/CFSA N, 2013).

13
Enteropathogenic – Escherichia coli

Lactose-fermenting species are generally harmless, although some strains cause gastrointestinal

illnesses. Consumption of pathogenic E. coli 0157 generated from infected meat causes colitis

with bloody diarrhoea, which can lead to haemolytic uremic syndrome consequences (Elizabeth

and Martin, 2013). E. coli is a major cause of diarrhoea in impoverished nations and areas with

poor sanitation. Indeed, it has been linked to "traveller diarrhoea". The most recent outbreak in

North America, however, happened in a nursing home in Ontario. Escherichia coli, Entero-

invasive, Haemorrhagic, and Enteropathogenic are the four sub-groups of enteropathogenic

bacteria. Each strain has unique characteristics; the main source of bacteria in the environment is

most likely infected human faces, but there may also be animal reservoirs and untreated water,

which are the most likely sources of food contamination. As a result of the 1993 E. coli 0157: H7

outbreak caused by contaminated hamburgers, the public became fully aware of E. coli 0157: H7

and its link to food. This outbreak sickened almost 700 people, and four children died as a result.

E. coli 0157: H7 can be obtained through the ingestion of undercooked meat, and it can be

transmitted from person to person via the faecal oral route. Infected people's diarrhoea faeces

may contain E. coli 0157: H7. Pathogens can spread if personal hygiene and hand washing

protocols are not followed (Buzby, 2011).

2.3 Lessons Learnt And Future Challenges

Salmonella, Campylobacter and strains of E. coli are well-established examples of common

food-borne pathogens. They are markedly different in terms of epidemiology, physiology,

ecology, host association and virulence properties, but together enable some generic conclusions

to be drawn on the overall persistence of food-borne bacterial disease over the last 20 years.

Although these are the major bacterial pathogens monitored, many others are also transmitted

14
through food. At anytime such relatively minor food-borne pathogens, like L. monocytogenes,

can also become major problems. Investigating the reasons for such shifts in patterns of food-

borne disease provides valuable information for future risk management strategies. From the

examples discussed above, for which data can be assessed over relatively long periods, it would

seem that there is little evidence we are winning significant ground in the battle against food-

borne illness caused overall by bacterial pathogens. In fact even the most successful

interventions, such as vaccination of chickens against Salmonella, have done little more than

reduce the pathogen load in the food chain. It can be speculated that such reductions in exposure

could have some adverse affects, for example, by altering the immune status of the population.

There are already disturbing trends such as shifts away from illnesses, such as

campylobacteriosis and listeriosis, in the young towards the increasingly growing older

population. In addition these pathogens are constantly evolving and adapting enabling the

exploitation of novel opportunities, for example, new vehicles created by modern processing

techniques, new retailing fads or new food consumption habits. This highlights the need for

multidisciplinary research, and especially the inclusion of social sciences, to investigate

changing trends in food-borne [Link] the substantial investments made by governments

and industry alike, these bacterial pathogens still feature as major public health problems.

However, these investments have not been without their successes; a combination of biosecurity

and vaccination has largely eliminated S. enteridis PT 4 from the breeder and layer flocks in

many European countries, while legislation and retailer education have sufficiently improved

hygiene to reduce E. coli 0157 in cooked meats. Clearly we need to share and implement

throughout Europe those strategies that are effective while maintaining constant vigilance against

15
the ability of such organisms to adapt to changing environments and to exploit the opportunities

that arise to occupy novel niches.

2.4 PRE-DISPOSING FACTORS TO FOOD-BORN ILLNESS

Biological Factors

Biological factors encompass microorganisms such as bacteria, viruses, parasites, and fungi that

contaminate food and cause illness. Bacterial pathogens like Salmonella, Escherichia coli, and

Listeria monocytogenes are common culprits in foodborne outbreaks. These pathogens thrive in

environments where food is mishandled or improperly stored, leading to contamination and

subsequent illness in consumers. Additionally, viruses like norovirus and hepatitis A can

contaminate food through infected food handlers or contaminated water sources. Parasites such

as Cryptosporidium and Giardia can also cause foodborne illness when food or water is

contaminated with fecal matter. Fungi, while less common, can produce toxins in food, leading

to illnesses like aflatoxin poisoning. (Boeing et al., 2012)

Environmental factors:

Environmental factors contribute significantly to the risk of foodborne illness. Poor sanitation

practices, inadequate food storage facilities, and contaminated water sources can all lead to food

contamination. For example, agricultural practices that involve the use of untreated manure or

contaminated irrigation water can introduce pathogens into the food supply chain. Furthermore,

natural disasters like floods or hurricanes can compromise food safety by contaminating crops

and disrupting food distribution networks. Climate change may also exacerbate these risks by

altering weather patterns and increasing the prevalence of foodborne pathogens in certain

regions. (Boeing et al., 2012)

16
Human-Related Factors

Human-related factors, including food handling practices, hygiene habits, and socioeconomic

status, play a crucial role in the occurrence of foodborne illness. Improper food handling, such as

cross-contamination between raw and cooked foods or inadequate cooking temperatures, can

introduce pathogens into food and lead to illness upon consumption. Moreover, poor personal

hygiene among food handlers, including improper handwashing and failure to adhere to food

safety protocols, can contribute to the spread of foodborne pathogens. Socioeconomic factors

such as poverty and lack of access to clean water and sanitation facilities also increase the risk of

foodborne illness, particularly in low-income communities. (Boeing et al., 2012)

Mitigation Strategies

Addressing the predisposing factors to foodborne illness requires a multifaceted approach that

involves collaboration between government agencies, food industry stakeholders, healthcare

professionals, and consumers. Key mitigation strategies include:

1. Strengthening Food Safety Regulations: Governments should enforce stringent food safety

regulations and standards to ensure compliance among food producers, processors, and

distributors. Regular inspections and monitoring of food establishments can help identify and

mitigate potential hazards.

2. Education and Training: Providing education and training programs for food handlers and

consumers on proper food handling, hygiene practices, and safe food storage can help prevent

foodborne illnesses. Empowering individuals with knowledge about food safety can reduce the

risk of contamination and illness.

17
3. Improving Sanitation Infrastructure: Investing in sanitation infrastructure, including access to

clean water and adequate waste disposal systems, is essential for maintaining food safety.

Improving hygiene conditions in food production and distribution environments can minimize

the risk of contamination.

4. Promoting Research and Surveillance: Continued research and surveillance efforts are crucial

for identifying emerging foodborne pathogens and monitoring trends in foodborne illness.

Surveillance systems can help detect outbreaks early and implement timely interventions to

prevent further spread.

2.5 Causes Of Food Borne Illnesses

majority of foodborne illnesses are caused by harmful bacteria and viruses. Some parasites,

fungi, Prions and chemicals also cause foodborne illnesses. (Boeing et al., 2015)

Bacteria

Bacteria are tiny organisms that can cause infections of the GI tract. Not all bacteria are harmful

to humans. Some harmful bacteria may already be present in foods when they are purchased.

Raw foods including meat, poultry, fish and shellfish, eggs, unpasteurized milk and dairy

products, and fresh produce often contain bacteria that cause foodborne illnesses. Bacteria can

contaminate food—making it harmful to eat—at any time during growth, harvesting or slaughter,

processing, storage, and shipping.

Foods may also be contaminated with bacteria during food preparation in a restaurant or home

kitchen. If food preparers do not thoroughly wash their hands, kitchen utensils, cutting boards,

and other kitchen surfaces that come into contact with raw foods, cross-contamination—the

spread of bacteria from contaminated food to uncontaminated food—may occur.

18
If hot food is not kept hot enough or cold food is not kept cold enough, bacteria may multiply.

Bacteria multiply quickly when the temperature of food is between 40 and 140 degrees. Cold

food should be kept below 40 degrees and hot food should be kept above 140 degrees. Bacteria

multiply more slowly when food is refrigerated, and freezing food can further slow or even stop

the spread of bacteria. However, bacteria in refrigerated or frozen foods become active again

when food is brought to room temperature. Thoroughly cooking food kills bacteria. (Askarji et

al., 2013)

Many types of bacteria cause foodborne illnesses. Examples include

• Salmonella, a bacterium found in many foods, including raw and undercooked meat, poultry,

dairy products, and seafood. Salmonella may also be present on egg shells and inside eggs.

• Campylobacter jejuni (C. jejuni), found in raw or undercooked chicken and unpasteurized milk.

• Shigella, a bacterium spread from person to person. These bacteria are present in the stools of

people who are infected. If people who are infected do not wash their hands thoroughly after

using the bathroom, they can contaminate food that they handle or prepare. Water contaminated

with infected stools can also contaminate produce in the field.

• Escherichia coli (E. coli), which includes several different strains, only a few of which cause

illness in humans. E. coli O157:H7 is the strain that causes the most severe illness. Common

sources of E. coli include raw or undercooked hamburger, unpasteurized fruit juices and milk,

and fresh produce.

• Listeria monocytogenes (L. monocytogenes), which has been found in raw and undercooked

meats, unpasteurized milk, soft cheeses, and ready-to-eat deli meats and hot dogs.

• Vibrio, a bacterium that may contaminate fish or shellfish.

19
• Clostridium botulinum (C. botulinum), a bacterium that may contaminate improperly canned

foods and smoked and salted fish.

20
2.6 Who Gets Food Borne Illnesses

Anyone can get a foodborne illness. However, some people are more likely to develop foodborne

illnesses than others, including

• infants and children

• pregnant women and their fetuses

• older adults

• people with weak immune systems

These groups also have a greater risk of developing severe symptoms or complications of

foodborne illnesses.(Scheule, 2001)

2.7 Symptoms Of Food Borne Illnesses

Symptoms of foodborne illnesses depend on the cause. Common symptoms of many foodborne

illnesses include

• vomiting

• diarrhea or bloody diarrhea

• abdominal pain

• fever

• chills

Symptoms can range from mild to serious and can last from a few hours to several days.

(Scheule, 2001).

C. botulinum and some chemicals affect the nervous system, causing symptoms such as

• headache

• tingling or numbness of the skin

• blurred vision

21
• weakness

• dizziness

• paralysis

2.8 The Complications Of Food Borne Illnesses

Foodborne illnesses may lead to dehydration, hemolytic uremic syndrome (HUS), and other

complications. Acute foodborne illnesses may also lead to chronic—or long lasting—health

problems.

Dehydration

When someone does not drink enough fluids to replace those that are lost through vomiting and

diarrhea, dehydration can result. When dehydrated, the body lacks enough fluid and electrolytes

—minerals in salts, including sodium, potassium, and chloride—to function properly. Infants,

children, older adults, and people with weak immune systems have the greatest risk of becoming

dehydrated.(Scheule, 2001).

Signs of dehydration are

• excessive thirst

• infrequent urination

• dark-colored urine

• lethargy, dizziness, or faintness

Signs of dehydration in infants and young children are

• dry mouth and tongue

• lack of tears when crying

• no wet diapers for 3 hours or more

• high fever

22
• unusually cranky or drowsy behavior

• sunken eyes, cheeks, or soft spot in the skull

Also, when people are dehydrated, their skin does not flatten back to normal right away after

being gently pinched and released.

Severe dehydration may require intravenous fluids and hospitalization. Untreated severe

dehydration can cause serious health problems such as organ damage, shock, or coma—a sleep-

like state in which a person is not conscious.

2.8.1 Treatment Of Food Borne Illnesses

The only treatment needed for most foodborne illnesses is replacing lost fluids and electrolytes to

prevent dehydration. Over-the-counter medications such as loperamide (Imodium) and bismuth

subsalicylate (Pepto-Bismol and Kaopectate) may help stop diarrhea in adults. However, people

with bloody diarrhea—a sign of bacterial or parasitic infection—should not use these

medications. If diarrhea is caused by bacteria or parasites, over-the-counter medications may

prolong the problem. (Scheule, 2001).

Medications to treat diarrhea in adults can be dangerous for infants and children and should only

be given with a health care provider’s guidance.

If the specific cause of the foodborne illness is diagnosed, a health care provider may prescribe

medications, such as antibiotics, to treat the illness.

Hospitalization may be required to treat lifethreatening symptoms and complications, such as

paralysis, severe dehydration, and HUS.

Eating Diet And Nutrition

The following steps may help relieve the symptoms of foodborne illnesses and prevent

dehydration in adults:(Marsus, 2015).

23
• drinking plenty of liquids such as fruit juices, sports drinks, caffeine-free soft drinks, and broths

to replace fluids and electrolytes

• sipping small amounts of clear liquids or sucking on ice chips if vomiting is still a problem

• gradually reintroducing food, starting with bland, easy-to-digest foods such as rice, potatoes,

toast or bread, cereal, lean meat, applesauce, and bananas

• avoiding fatty foods, sugary foods, dairy products, caffeine, and alcohol until recovery is

complete Infants and children present special concerns. Infants and children are likely to become

dehydrated more quickly from diarrhea and vomiting because of their smaller body size. The

following steps may help relieve symptoms and prevent dehydration in infants and children:

• giving oral rehydration solutions such as Pedialyte, Naturalyte, Infalyte, and CeraLyte to

prevent dehydration

• giving food as soon as the child is hungry

• giving infants breast milk or fullstrength formula, as usual, along with oral rehydration

solutions Older adults and adults with weak immune systems should also drink oral rehydration

solutions to prevent dehydration.

Food Hygiene

Food hygiene is described as a sanitary science that strives to produce food that is safe for human

eating and of good storage quality and it includes any sanitation procedures that prevent bacteria

and other germs of human origin from reaching food (Umoh and Odibo, 2009). Food hygiene is

a broad topic that entails researching strategies for producing and preparing food that is both safe

and of high quality. It includes not only proper handling of all types of foods and beverages, but

also food contact surfaces such as utensils and apparatus used in the preparation, service, and

consumption of the food, as well as precautions to avoid contamination with food poisoning

24
bacteria that may come from the animal or part plant host supplying the food (Umoh and Odibo,

2009).

2.8.2 Factors That Contribute To Food Borne Illness

Poor Personal Hygiene

Poor personal hygiene can lead to food contamination, such as when certain food vendors fail to

properly wash their hands after using the restroom or toilet, posing a major danger of faecal

contamination (FDA, 2014). Everyone has germs on their skin, lips, hands, and a variety of other

organisms on other areas of their bodies, such as their hair. Food-borne disease can be caused by

food service staff contaminatingfood. Pathogens can be transmitted to food by food workers

from a contaminated surface, from one item to another, or from hands contaminated with

organisms from the gastrointestinal tracts (Munide and Kuria, 2015). As a result, hand contact

with ready-to-eat food, i.e. food that is edible without washing, cooking, or additional

preparation by the consumer or the food establishment and is expected to be consumed in that

manner, represents a potentially important mechanism through which pathogens may enter the

food supply (Munide and Kuria, 2015).

Abuse Of The Time –Temperature Relationship

Another aspect that might lead to food-borne infections is the misuse of the timetemperature

relationship. Controlling the time that food is in the temperature risk zone is critical for

preventing food-borne illness; this means that hot meals should be maintained at 140oF or above,

while cold foods should be kept at 41oF or lower (FDA, 2014). Cooked or chilled items, such as

salads, should not be left at room temperature for more than two hours (FDA, 2014). Problems

with time-temperature relationships arise as a result of:

25
1. Food is not stored, prepared or held at the required temperature; food is not cooked or reheated

to temperature high enough to kill harmful micro organisms

2. Food is prepared in advance of service and proper temperature control is not maintained

(FDA, 2014).

Cross-Contaminating Raw And Cooked Food

Cross-contamination of raw and cooked food refers to the transfer of potentially hazardous

bacteria from a surface to food or from one food to another. When food contact surfaces are not

cleansed or sterilized as required for food safety, cross contamination can occur (FDA, 2014). To

avoid cross-contamination, wash your hands with soap and warm water before beginning to

prepare food, before handling a different item (for example, if you have touched raw chicken,

wash your hands before preparing a salad), and after using the restroom. Sneeze or cough away

from food. Organisms can "travel" from raw to cooked food, so never allow raw food to come

into contact with cooked food (FAD, 2014).

2.8.3 Prevention And Control Of Food Borne Illnesses

Prevention and control of food–borne diseases, regardless of the specific cause, are based on the

same principles:

1. Avoidance of food contamination

2. Destruction or prevention of contaminants

3. Prevention of further spread or multiplication of contaminants.

Specific modes of intervention vary from area to area depending on environmental, economic,

political, technology and socio cultural factors. The preventive and control strategies may be

approached based on the major site in the cycle of transmission or acquisition where they are

26
implemented. These involve the activities performed at three different stages, which are as

follows;

1- Source of infection

2- Environment and

3- Host.

1. Source of infection

✓ Thorough cooking of raw.

✓ Thorough washing of raw vegetables with clean water

✓ Inspection of food

✓ Treatment of cases

✓ Washing hands, knives, cutting boards, etc. after handling uncooked foods.

✓ Avoiding contact with materials contaminated with pet excreta or soil.

✓ Washing hands after contact with animals

✓ Management of food handlers and homemakers

✓ Treatment of carriers.

✓ Proper care for patients with food-borne illnesses.

✓ Avoidance of food from animals with obvious infection, e.g., mastitis in cows

✓ Treatment of infections in food handlers such as skin and throat infections

2. Environment

This involved stringent follow-up from production to consumption. Some of the interventions

include:

✓ Freezing, salting, etc. of food items during storage

✓ Control of flies, rats, roaches

27
✓ Public education on environmental and personal cleanliness

✓ Surveillance of food establishments

✓ Avoiding contamination of food after cooking.

✓ Maintenance of sanitary food area.

✓ Proper handling

3. Host

✓ Active or passive immunization of susceptible hosts

✓ Health education on the above areas.

2.9 “Some Golden Rules” Of Who For Safe Food Preparation (10)

1. Choose foods processed for safety

2. Store foods carefully

3. Avoid contact between raw and cooked food

4. Wash hands repeatedly

5. Protect food from insects, rodents and other animals

6. Use safe water

28
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Study Area

This research project is focused at Wudil Market, Wudil l.g.a. Kano State, which accounts a

population of thousands of people's for which 10% of their demands for daily consumption

depend on these selected vendors.

3.2 Collection Of Samples

One (1) sample each of Watermelon (Citrullus lanatus) and Carrot (Daucus carota) were

collected randomly from 4 different stationery vendors in Wudil Market. This makes a total of 8

samples collected on August, 2024. The samples was collected in a sterile polytene bag to avoid

contamination and were transported immediately to the laboratory for analysis.

3.3 Sterilization Of Materials

All materials used were adequately and appropriately sterilized before and after use. Glass wares

such as test tubes, conical flasks, pipettes, etc were thoroughly washed with detergents, rinsed

properly with water and drained. They were wrapped in aluminum foil and sterilized in hot air

oven at 170ºC for 1 hr. Metal equipments like the inoculating loop were heated to redness in an

open flame before and after use. The laboratory bench was always swabbed using 70% alcohol

for disinfection before analysis was made. Every isolation and inoculation was done near the

flame to reduce contamination of the agar plates tubes.

3.4 Preparation Of Culture Media

All media (Nutrient agar, MaConkey agar, Eosin methlenyne blue agar,Salmonella Shigella agar,

and Manitol salt agar, used for culturing were prepared according to standard specification by the

29
manufacturer and were sterilized at 121 ℃ for 15 minutes.

3.5 Preparation Of Sample

Ten grams (10g) of each sample was measured with the aid of a weighing balance, it was pressed

using a sterile pestle and transferred into a sterile conical flask. A steriled syringes was used to

obtain 9ml of sterile distilled water and this was introduced into the conical flask.

3.6 Seriel Dilution

A test tubes containing exactly 9ml of distilled water was covered with cotton wool and

Sterilized in an autoclave at 121°C for 15 minutes. The tubes are allowed to cold and stock

Solutions was prepared. The tubes was Labelled as 10–¹, 10–², 10–³,10–⁴, and 10–⁵. Using

steriled syringe 1ml from stock solution was transferred into 10–¹, 10–¹ to 10–², 10–² to 10–³, 10–

³ to 10–4,. 10–⁴ to 10–5 and sparks Gently (Kaur and Rat, 2015).

3.7 Isolation And Enumeration Of Bacteria

About 0.1ml of 10⁻³, 10–⁴, and 10–5 dilution for each sample was inoculated on the different

solidified and sterilized agar plates. The inoculums on each plate was spread using a steriled wire

loop and the plates were inverted and placed in an incubator for 24-48 hours at 37 ℃.

Enumeration of total bacteria count was done using the plate count method, colonies present

were counted and recorded to get the total colony count in cfu/ml.

30
3.7.1 Sub- Culture Technique

Some bacterial isolates were selected and inoculated in a petri dishes containing various

differential and selective media. The plates are then incubated at 37⁰c for 24 hrs. After

incubation, pure colonies was obtained.(Karoki et al., 2018)

3.8 Enumeration Of Coliform Bacteria

Presumptive Test

After seriel dilution was prepared. 2- 3ml each From 10–¹, 10–² and 10–³ dilution was measured

using steriled syringes. 1ml of each dilution was inoculated into 3 test tubes Containing lactose

broth and inverted Durham tubes. The LST test tubes was incubated at 37°c for 24 [Link]

production (bubbles) and acid formation was observed. The test tubes showing gas production

was recorded.

Confirm Test For Coliform

A Loopful From each gas positive test tubes of the LST was transfered to a separate test tubes of

BGLB broth. The BGLB test tubes was incubated at 37°C for 45hrs and observed for gas

formation.

Completed test

The test organism was Inoculated on a selective agar medium (e.g., E. coli agar,

MacConkey agar) with the diluted sample and incubated the agar plates at 35°C ±

0.5°C for 24-48 hours. The number of colonies on the agar plate was counted.

3.9 Identification Of Bacterial Isolates

Identification of bacterial isolates was done using Gram stain technique and biochemical test

such as catalase, indole, citrate utilization, oxidase, methyl red and voges- proskauer test.

31
Making Smear

Using sterilized wire loop a small portion of the growth was transferred to the glass Slides

containing a drop of distilled water. It was emulsified on the distilled water until a homogeneous

solution is formed and the film was allowed to dry by passing it over a burnsen flame.

Gram Staining

1 or 2 drops of Crystal violet was added and allowas for 30-40 [Link] glass slide was

drained up and washed with water. Lugol's lodine was added and allowed for 60 seconds. The

micascope glass slide was drained up and washed with water gently. The microscope glass slide

was decolonized with acetone alcohol until it appeared free of Violet Strain and. 2-3 drop of

safranim was added. Microscopic glass slide was rinsed with water and a drop of oil immesion

was applied and examine with oil immersion objectves Lens of the light -microscope.

Catalase Test:

Three (3) % H₂O₂ was introduced unto a clean grease free slide. A smear of loop full bacteria

was made. Formation of bubbles was observed for each bacterial isolate (Mahon, 2011).

Indole Test:

Peptone broth was prepared by weighing (15) ml of peptone broth in a test tube and sterilized in

an autoclave at 121℃ for 15 minutes at 15 ibs pressure. A loop full of bacteria culture was

inoculated in broth and incubated for 24-48 hours at 37 ℃. Two (2) drops of Kovac’s reagent

was dispensed into the test tubes and mixed together after sterilization (Abiola and Oyetayo,

2016).

Citrate Utilization Test:

Two (2) grams of Sodiun Citrate, 5g Sodium Chloride, 1 g Dipotassium Phosphate, 1 g

Ammonium Dihydrogen Phosphate, 0.08 g Bromothylmol Blue, 0.2 g Magnesium Sulphate and

32
15g agar was mixed together and 1000ml sterile distilled water was dispensed in the same

mixture. The pH was adjusted to 6.9 and gentle heat was applied to dissolve agar. About 3-4 ml

was collected in glass bottles and sterilized at 121 ℃ for 15 minutes in an autoclave. This was

cooled in a slant bottles and inoculums was smeared onto the surface of the slant (Chester and

Copper, 2019).

Methyl Red Test

The bacteria culture was inoculated into a fresh sterile broth medium and incubated at 37 ℃ for

48 hours. A sterile pipette was used to dispense 5 drops of Methyl red reagent into the broth

culture and colour change was observed (McDevitt, 2009).

Voges- Proskauer Test:

Preparation of 5 % a – naphthol in ethanol and 40 % Sodium hydroxide in deionized water was

done. MR-VP broth was also prepared and 5 ml dispensed in different test tubes and sterilization

was done at 121 ℃ for 15 minutes using an autoclave. The medium was allowed to cool to room

temperature. Inoculum from fresh culture media was introduced in different test tube and this

was incubated together with the control at 37 ℃ for 48 hours. About 2.5 ml of culture was

dispensed in a sterile cultures tube and 5 drops of methyl red reagent was added. The test

organism was also compared with the control and colour change was observed (McDevitt, 2009).

Coagulase Test

Few drops of physiological saline was placed on two separate grease free slide and a loop of

bacterial isolate was emulsified on the slide to make two suspensions. A drop of human plasma

was collected with a sterile Pasteur pipette and mixed gently on the slides. The two slides were

observed for clumping between 5-10 minutes for positive result (McAdow et al., 2012).

Urease test

33
The urease test medium was prepared as instructed by manufacturer. Using a steriled wire loop

the test organism was inoculated in a test tubes Containing 5ml of urease broth and incubated at

37⁰c for 24hrs. The medium was observed for a color change after 24 hours.

3.9.1 Data Analysis

Results obtained was subjected to suitable statistical analysis (e g Enumeration of bacterial count

was done using colony forming unit per millimeter (cfu/ml).

34
 CHAPTER FOUR

4.0 RESULT

4.1 Total Bacterial Count :

Bacterial load of the watermelon and carrot varied with type and vendor (Table 1). Watermelon

from Vendor one (W-1) had the lowest bacterial load (0.90 x 10–¹ cfu/ml) of all the samples,

while watermelon from vendor four (W-4) had the highest bacterial load (1.30 x 10¹ cfu/ml)

(McDevitt, 2009).

Table 4.1: Total Bacterial count from watermelon and carrot (x 10⁴ cfu/ml)

S/No. SAMPLES Total bacterial count (cfu/ml)

1 W-1 0.90×10–¹

2 W-2 1.10×10–¹

3 W-3 1.00×10–¹

4 W-4 1.30×10–¹

5 C-1 1.00×10–¹

6 C-2 1.00×10–¹

7 C-3 1.11×10–¹

8 C-4 1.14×10–¹

KEY: W1-4 Bacterial isolates on watermelon and C 1-4; Bacterial isolates on carrot

35
4.2 Total Coliforms Count

The bacterial count for coliforms on all samples varied with type and vendor. Watermelon from

vendor four (W-4) and carrot from vendor one (C-1) showed the presence of coliform while the

others had no coliform . (Table 2).

Table 4.2: Coliform Total count of watermelon and carrot

S/NO. Sample Coliform count (MPN/g)

1 W-1 3

2 W-2 3

3 W-3 3

4 W-4 <3

5 C -1 <4

6 C-2 3

7 C-3 3

8 C-4 3

W 1-4; Coliform isolates on watermelon and C 1-4; Coliform isolates on carrot

36
4.3 Gram Stain Reaction:

Two (2) isolates were Gram positive bacterial while six (6) isolates were Gram negative bacteria

with the characteristics colour of Purple for Gram positive and Pink for Gram negative (Table 5)

4.4 Morphological Characteristics of Bacterial Isolates:

Based on the morphological characteristics of bacterial isolates observed on the

selective/differential media (Table 5); Isolates on MaConkey appeared Smooths, Opaque and

Motile. Isolates on Eosin Methylene Blue agar appeared Pink, Mucoid, and some with green

metallic sheen. Manitol Salt agar developed colonies that were yellowish and oily. A blackish

substance was found on Salmonella Shigella agar with the organism appearing circular and

transparent in the middle of the black substance.

Table 4.5: Morphological Characteristics of Bacterial isolate on Selective/Differential

Media

Plates Colour Size Elevation Shape Gram's reaction

WA-1 Yellow Small Raised Cocci +Ve
WA-2 Pink Small Umbonate Rod -Ve
WA-3 Green Small Flat Rod -Ve
WA-4 Opaque Large Flat Cocci -Ve
CB-1 Yellow Small Flat Cocci +Ve
CB-2 Black Small Raised Rod -Ve
CB-3 Translucent Medium Raised Rod -Ve
CB-4 Green Small Flat Rod -Ve
(-ve) - negative; (-ve) - positive; WA 1-4; Bacterial isolates on watermelon from different culture

media. CB 1-4; Bacterial isolates on carrot from different culture media.

37
4.5 Biochemical reaction of the Bacterial Isolates.:

Biochemical test confirmed the presence of Staphylococcus spp., Proteus spp., Salmonella spp.,

Escherichia spp., and Shigella as stated on the row for probable organism (Table 6).

Table 4.6: Biochemical Reaction of Bacterial from watermelon and carrot

ISOLATE CA CO IND URE CIT MR VP Probable Organism

S
WA-1 + + - + + + + Staphylococcus spp.
WA-2 + + - + + + + Staphylococcus spp.

WA-3 + - + - - + - E. coli app.

WA-4 + - + + + + - Proteus spp.
CB-1 + + - + + + + Staphylococcus spp.

CB-2 + - - - - + - Salmonella spp.

CB-3 + - - - - + - Shigella spp.

XB-4; + - + - - + - E. coli spp

*(+ve), positive; (-ve), negative; CA, catalase; CO, coagulase; IND, indole; URE, urease, CIT,

citrate; MR, methyl red; VP, vogeproskauer.

WA 1-4; Bacterial isolates on watermelon

CB 1-4; Bacterial isolates on carrot

38
4.6 Discussion

Results from this study show the presence of harmful bacteria in watermelon and carrot samples

purchased from Wudil Market, Wudil L.G.A, Kano state.. The isolated organisms were;

Escherichia spp., Staphylococcus spp., Salmonella spp., Proteus spp., and Shigella spp. The

highest level of bacterial load was recorded on carrot (Table 1). Cross contamination of fresh

produce could have been from the vendor or the environment since the operating premises is

usually kept unclean. The previous study of Akusu et al., (2016) on vegetable salads from street

foods among different vendors in Port Harcourt metropolis in Nigeria agrees with the present

study as high bacterial load was observed in some of the selected [Link] in the number of

human infections and outbreaks is a resultant effect of the high rate in consumption of fruits and

vegetables (Mashak et al., 2015) as most pathogenic or opportunistic bacteria inhabit them (Berg

et al., 2014). Contamination of fruits and vegetables by spoilage organism or harmful bacteria

usually occur at any stage of production to the consumer (Berg et al., 2014). Fruits and

vegetables have there microflora which are often yeast, molds and spoilage bacteria, it was also

discovered that they can harbor harmful bacteria such as Escherichia coli, Salmonella, Shigella,

Listeria, Campylobacter, Monocytogenes, Yersinia enterocolitica, Clostridium botulinum,

Bacillus cereaus as well as parasite (Mritunjay and Kumar, 2015). Microbes that are non-

pathogenic are found to increase the rate of spoilage hereby diminishing the quality of the

produce and reduction in market value. Since fruits and vegetables harbours microorganisms

they also help in spreading microbes from one area to other areas food is being prepared (Altieri

and Nicholls, 2017). The presence of coliforms on some samples (Table 2) could be a result of

feacal contamination or an indication of poor sanitary of the vendors (Oje et al., 2018). The

presence of Escherichia, Staphylococcus, Salmonella, and Shigella is of great concern in public

39
health since most of these microorganisms are virulent. Olawale et al., (2015) reported different

prevalence of virulent genes in Enterococcus feacalis isolated from ready to eat foods.

Foodborne illnesses are a growing public health distress, social disturbance, avoidable economic

burden and preventable death. One of the normal flora of the human and animal intestine is

Esherichia coli often known as enteric bacteria is one of the common cause of foodborne illness

in the world (Sharff, 2012). Salmonella spp. Should not be found in 25 g of ready to eat fruits

and vegetable meant for human consumption and therefore must be rejected (Food and Drugs

Board, 2013; Abakari and Cobbina, 2018). The presence of Salmonella on the carrot sample

depicts that the vegetable is unfit for human consumption going by the guidelines. Shigella spp.

Isolated on both watermelon and carrot samples is unsatisfactory for human consumption

according to health protection agency, 2019 that states that food containing Shigella in about 25

g of sample is unsafe for human consumption (Abakari and Cobbina, 2018). The fruits and

vegetable sample contaminated with Shigella spp. could be as a result of improper hygiene

practices by vendors in Oja-Oba market, Ilorin. The level of hygiene practices can influence the

total number of bacterial load on fruits and vegetables. The source where the fruits and vegetable

are gotten could have influenced the high number of bacterial load in selected samples. Several

factors in each collection point could have contributed to contamination on samples. Data

analysis of mean bacterial count isolated revealed. Bakobie et al., 2017 reported that there was

no significance difference in pathogenic bacteria countdone on fruits and vegetable sample from

different vendors in Wudil Market. This study has identified the presence of Escherichia,

Salmonella, Proteus, Shigella and Staphylococcus on selected fruits and vegetable sample

obtained from the Wudil Market, Wudil L.G.A., Kano state. In conclusion, the present study has

shown that watermelon and carrot sold by street vendors in Wudil market are not safe for human

40
consumption and consumers are at health risk in terms of microbial quality. Contamination from

farms or production area, Improper food handling while processing, non-hygienic practices while

packaging and environmental conditions are major factors responsible for high microbial load on

fruits and vegetables. This study shows that there is an urgent need for regulation agency to vet

food vendors in other to promote improvement in quality standards and food safety of ready-to-

eat foods in Wudil Market metropolis.

41
 CHAPTER FIVE

5.0 SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

5.1 Summary

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. Reliable identification of bacteria remains to be an

important task in food microbiology. Isolation and identification of bacteria was carried out on

watermelon and carrot sold in Wudil Market, Wudil L.G.A Kano State. In this study, eight (8)

samples of watermelon and carrots were collected randomly from different stationery vendors

and analyzed immediately through serial dilution, inoculation, incubation, subculture,

microscopy and biochemicals to confirm the isolated organisms. A total of five pathogenic

bacteria were isolated: Staphylococcus aureus (42.3%), Escherichia coli (40.8%), Salmonella

(5.6%), Shigella (4.2%) and Proteus spp.(4.2%). This indicates 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 sold in Wudil

Market and thereby heighten the safety of consumer.

5.2 Conclusions

This study has demonstrated that some of the most popular types of ready-to-eat fruits and

vegetables that are sold in Wudil Market, Wudil L.G.A., Kano state, are contaminated, and do

not meet the required quality and safety levels. Some of the isolated bacteria especially

Staphylococcus aureus and Escherichia coli that are isolated in almost each and every collection

of the samples are potential enteric pathogens and are known to cause gastroenteritis. This

clearly depicts poor handling and management leading to cross contamination to [Link] a

42
normal flora of the skin and [Link] demonstrate feacal contamination. This pose a health threat to

the patron and efforts to reduce level of contamination in this market and are highly

recommended as not only people from Wudil rely on the fruits and vegetables, but also some

individuals from other communities.

5.3 Recommendations

1- It is recommended that food and drug authority should ensure that the street vendors are

educated on good and proper hygiene while processing fresh produce for human consumption

and enforce strict [Link] policy developed has to respond to an integrated consultation

with vendors and consumers if it is to meet the needs of each of the partners in food safety

(government, school management, consumers and vendors). 2- There is a need for further study

on foods classified as medium to high risk that include E. coli, Salmonella and other food-borne

microorganisms. Determinations of these types of bacteria act as a quick index of acceptability

of food for human consumption.

43
REFERENCES

Abdolmaleki, Z., Mashak, Z. and Dehkordi, S. (2019).Phenotypic and genotypic characterization

of antibiotic resistance in the methicillin-resistant Staphylococcus aureus strains isolated
from hospital cockroaches. Antimicrobial Resistance and Infection Control, 8 (54): 2-14.

Adegoke, A. A. and Okoh, I. A. (2013). Staphylococcus species and emerging traits in their
commensal subgroup: A Call to Arms. Journal of Pure and Applied Microbiology, 7(4):
3281-3291.

Adegoke, A.A. and Okoh, A.I. (2014). Species diversity and antibiotic resistance properties of
Staphylococcus of farm animal origin in Nkonkobe Municipality. Folia Microbiology,
59:133-140.

Adegoke, A.A. and Okoh, A.I. (2015). Antibiogram of Stenotrophomonas maltophilia isolated
from Nkonkobe Municipality, Eastern Cape Province, South Africa. Jundishapur Journal
of Microbiology. 8(1): e13975.

Adegoke, A.A., Faleye, A.C., Singh, G. and Stenström, T.A. (2016). Antibiotic Resistant
Superbugs: Assessment of the Interrelationship of Occurrence in Clinical Settings and
Environmental Niches. Molecules, 22(1): 29;

Akinduti, P. A., Ejilide, O., Motayo, B. O and Adeyokinu, A. F.(2012). Emerging multi-drug
resistant. AmPC betalactamase and carbepenase enteric isolates in Abeokuta, Nigeria.
Nature and Science, 10(7): 70-74.

Becker, K., Heilmann, C. and Peters, G. (2014). CoagulaseNegative Staphylococci. Clinical

Microbiology Reviews,27: 870–926.

Ben, M., Cark, Y. and Rik, G. (2017). Staphylococcus aureus isolated from wastewater treatment
plants in Tunisia: occurrence of human and animal associated lineages. Journal of Water
Health, 65: 23-36.

Cassat, J., Lee, C. and Smeltzer, M. (2007). Investigation of biofilm formation in clinical isolates
of Staphylococcus aureus. Review of Methods in Molecular Biology, 391:127–134.

Clinical and Laboratory Standard-CLSI. (2015).Performance Standards for Antimicrobial

SusceptibilityTesting; Twenty-Fifth Informational Supplement. In: CLSI document
M100- S25. Wayne.

Cramton, S., Gerke, C., Schnell, N., Nichols, W. and Götz, F. (1999). The intercellular adhesion
(ICA) locus is present in Staphylococcus aureus and is required for biofilm formation.
Infection and Immunity, 67:5427–33.

Dehkordi, S., Gandomi, H., Akhondzadeh-Basti, A., Misaghi, A. and Rahimi, E. (2017).
Phenotypic andgenotypic characterization of antibiotic resistance of methicillin-resistant

44
 Staphylococcus aureus isolated from hospital food. Antimicrobial Resistance and
InfectionControl, 6:104-107.

Donkor, E. S. (2019). Nosocomial Pathogens: An In-Depth Analysis of the Vectorial Potential of

Cockroaches. Tropical Medicine and Infectious Disease, 4: 14-24.

Evans, J. and Wittler, M. (2019). Amoxicillin Clavulanate. StatPearls Publishing, Treasure

Island. Gary, M and Durben, L. (2004). Medical and vertinary entomology. Medical and
Veterinary Practices News, 597: 204 – 285.

Gehad, T. E and Emman, T. E (2011). The role of cockroaches and flies in mechanical
transmission of medical important parasites. Journal of Entomology and Nematology, 3
(7) : 98 – 104.

Gray,M. and Durden, L. (2004). Medical and Veterinary entomology. Medical and veterinary
pratices news, 597:04 –85.

Harwood, R. and James, M. (1979). Entomology in human and animal health. 7th Edition. New
York, Macmillan Publishing.

Holt, J. G., Krieg, N. R., Sneath, P. H. A., Stately, J. T. and Williams, S. T. (1994). St. Bergey’s
Manual of Determinative Bacteriology (9th Edition). Baltimore, Williams and Wilkins.
787p Ifeanyi, O. (2015). Microbiology of Cockroaches – A Public Health Concern.
International Journal of Scientific Research, 4 (4): 22-34.

Melton, L. (2012). Mechanical vectors of bacterial GI diseases. Acta tropica, 78: 3 – 17.

Menasria, T., Moussa, F., El-Hamza, S., Tine, S., Megri, R. and Chenchouni, H. (2014).
Bacterial load of German cockroach (Blattella germanica) found in hospital environment.
Pathogens and Global Health, 108: 141–147.

Pai, H., Chen, W. and Peng, C. (2005). Isolation of bacteria with antibiotic resistance from
household cockroaches (Periplaneta americana and Blattella germanica). Acta Tropica,
93: 259 –265.

Pollianna, S., Souza, G., Campos, B., Yatsuda, R. and Marques, L. (2014). Isolation,
pathogenicity and disinfection of Staphylococcus aureus carried by insects in two public
hospitals of Vitória da Conquista, Bahia, Brazil. The Brazilian Journal of Infectious
Diseases, 18(2) :129–136.

Regenthal, P., Hansen, J.S., André, I. and LindkvistPetersson, K. (2017). Thermal stability and
structural changes in bacterial toxins responsible for food poisoning. PLoS One,
12(2):e0172445.

Tsutsuura, S. and Murata, M. (2017). Temperature dependence of staphylococcal enterotoxin A

production by Staphylococcus aureus. Nihon Rinsho. 70(8):1323-1328.

45
Valles, S. M., Koehler, P. G. and Brenner, R. J. (1999).Comparative insecticide susceptibility
and detoxification enzyme activities among pestiferous btattodea. Compartive
Biochemistry and PhysiologyToxicology.124(3): 227–232

46