FEDERAL UNIVERSITY DUTSE(FUD)

ASSESSMENT OF THE BACTERIAL LOAD OF READY-TO-EAT FRUITS

SOLD BY STREET VENDORS IN DUTSE METROPOLIS JIGAWA STATE

MCB(499)

A PROJET SUBMITTED TO MICROBIOLOGY AND BIOTECHNOLOGY DEPARTMENT,

FACULTY OF LIFE SCIENCE, IN PARTIAL FULFILLMENT FOR THE REQUIREMENT

OF AWARD IN (BSC) DEGREE IN MICROBIOLOGY.

BY

MUSBAHU LAWAL

FSC/MCB/20/1083

SUPERVISED

BY

Mr. UMAR N. MUHTAR

NOVEMBER. 2025
 DECLARATION

I MUSBAHU LAWAL with registration number FSC/MCB/20/1083 hereby declared that this

Projet research titled, ASSESMENT OF BACTERIAL LOAD OF READY TO EAT

FRUITS FROM STREET VENDORS IN DUTSE METROPOLIS, is my own original work,

done by me under the guidance of Mal. UMAR N. MUHTAR and this work is submitted in the

partial fulfillment of the requirement for the award of Bachelor of Science in Microbiology. I

have acknowledged all sources of information and have ensured that any work taken from other

sources has been properly referenced and attributed. This work complies with the University’s

policies on academic integrity and ethics.

……………………………….. ………………………….

Sign Date
 CERTIFICATION

This Projet research was carried out at Federal University Dutse, Faculty of Science, Department

of Microbiology by MUSBAHU LAWAL with the registration number FSC/MCB/20/1083 and

supervised by Mal. UMAR N. MUHTAR and it is found to meet the essential requirements of

the report writing in the department of Microbiology, Federal University Dutse.

Mr. UMAR N. MUHTAR ________________________

(SUPERVISOR) SIGNATURE & DATE

DR. HINDATU YUSUF _______________________

(HEAD OF DEPARTMENT) SIGNATURE & DATE

MAL. LAWI ABDULLAHI _________________________

(PROJECT COORDINATOR) SIGNATURE & DATE

Prof. _________________________

(EXTERNAL SUPERVISOR) SIGNATURE & DATE

 DEDICATION

This Project work is dedicated to my Family especially my parents for their unwavering and

selfless support, and also those who help and contributes through my academic journey in one

way or the other.

 ACKNOWLEDGEMENT

I wish to extend my profound gratitude to my loving parents that supported and encouraged me

through happy and trying times, their endless prayers kept me going serving as a drive to

achieving my goals. Sincere appreciation goes to my supervisor, Mal. UMAR N. MUHTAR and

lecturers in my department who theoretically laid the foundation of my experiences. My

thankfulness goes to friends and dear ones, well-wishers and everyone that went above and

beyond one way or the other that has contributed to ensure this work was achieved.
 CHAPTER ONE

1.0 Introduction

Ready-to-eat fruits are conveniently prepared by street vendors for immediate consumption

without the need for washing or peeling. They offer a quick, healthy snack option rich in

essential nutrients like vitamins, fiber, and antioxidants. Regular consumption helps prevent

deficiencies and reduces the risk of chronic diseases, making them a valuable part of a

balanced diet. (Boampong et al., 2023). Although ready-to-eat fruits and fresh fruit juices are

convenient and widely consumed, they may present notable public health risks when handled

without proper hygiene. Contamination typically arises from exposure to contaminated

environments, poor personal hygiene among vendors, and the use of unsafe water or unclean

cutting tools. Inadequate washing of fruits, unhygienic display surfaces, contact with flies, and the

absence of refrigeration all encourage microbial growth. Such conditions can facilitate the

transmission of foodborne diseases, particularly gastrointestinal infections, caused by pathogens

such as Escherichia coli O157:H7, Salmonella spp., Shigella spp., and Staphylococcus aureus. (Ray et

al., 2024). While food safety has long been a global concern, modern practices differ widely

between developed and developing nations. In industrialized countries, advanced methods of

food processing and packaging help ensure safety. In contrast, many underdeveloped and

developing countries still rely on traditional methods, especially for fresh produce and ready-

to-eat foods. These items are often sold in open-air environments, exposed to poor hygiene

and harsh [Link] widespread consumption of minimally processed or inadequately

prepared food in these regions contributes significantly to the spread of parasitic infections.

Conversely, unsafe agricultural practices—such as improper harvesting methods, careless

handling of produce, and the use of contaminated water or untreated manure—compromise

the microbial safety of fruits and vegetables in many parts of the world. In their natural

environment, these crops are vulnerable to contact with a variety of microorganisms,

increasing the risk of contamination before they even reach the market. Food safety concerns

have led to the establishment of regulatory bodies that set standards to ensure food is safe for

consumption. One such body, the Codex Alimentarius, has developed globally accepted

guidelines that serve as a benchmark for many countries to maintain food quality and safety.(

Boampong,el al,.2023).

1.1 Statement of Research Problem

The consumption of ready-to-eat (RTE) fruits sold by street vendors is a widespread practice

in many urban centers of developing countries, including Dutse metropolis. These fruits

offer convenience and nutritional benefits to consumers, especially in fast-paced urban

environments. However, their safety is often compromised due to poor handling practices,

lack of regulatory oversight, and unhygienic environmental conditions in which they are

prepared and [Link] contamination is often linked to improper washing, use of

contaminated water, exposure to dust and flies, and poor personal hygiene of vendors.

( Sornka et al., 2024)... In Dutse metropolis, like many other growing urban areas in Nigeria,

street vending is largely unregulated, and RTE fruits are commonly sold under substandard

sanitary conditions. Despite the potential health risks posed by these contaminated fruits,

there is limited local data on their microbial quality in Dutse. Without concrete evidence and

assessment of the bacterial load on these products, public health authorities lack the basis for

creating effective policies or interventions. Therefore, this study seeks to assess the bacterial

load of ready-to-eat fruits sold by street vendors in Dutse metropolis, providing critical

insight into their safety and potential health implications.

1.3 Justification.

Ready-to-eat fruits are widely consumed in Dutse due to their affordability and

convenience, but poor hygiene and lack of regulation in street vending raise serious public

health risks. Inadequate handling practices can lead to microbial contamination, highlighting

the need for local research to guide food safety measures. (WHO 2015). Numerous studies

conducted in other parts of Nigeria and beyond have reported high microbial loads and the

presence of foodborne pathogens in RTE [Link], There is a lack of specific data on

the microbial safety of ready-to-eat fruits in Dutse metropolis, making it difficult for

authorities to manage health risks effectively. This study seeks to fill that gap by evaluating

bacterial contamination levels, providing evidence to support better hygiene practices,

inform public health policies, and guide future food safety regulations. Its findings could

also serve as a valuable reference for other cities facing similar challenges.

2.0 Aim and Objectives

2.1 Aim: To assess Bacterial load and presence of Escherichia coli and Staphylococcus

aureus in ready-to-eat fruits sold by street vendors in Dutse metropolis.

2.2 Objectives:

1) To determine the bacterial load and presence of Escherichia coli and Staphylococcus

aureus in selected ready-to-eat fruits sold by street vendors in Dutse metropolis.

2) To compare the levels of contamination among different fruit types and vending locations.

3) To assess the hygienic practices of street fruit vendors that may influence microbial

contamination.
 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Composition of Fruits.

Fruits provide vital nutrients—including vitamins, minerals, and easily digestible sugars—

essential for human growth, metabolism, and overall health. Their regular consumption helps

prevent nutritional deficiencies, lowers the risk of chronic diseases, and supports the body’s

detoxification processes. Fruits are commonly sold either whole or as fresh-cut varieties, with

over twenty types globally marketed in minimally processed forms for convenient

[Link] are vital components of a healthy diet, consisting primarily of water (about

80–90%), carbohydrates, organic acids, vitamins, minerals, and phytochemicals (Oyedele et al.,

2020). These constituents vary depending on the fruit species, degree of ripeness, and

environmental factors during cultivation and storage. Fruits are natural powerhouses of essential

vitamins crucial for maintaining health. They provide vitamin C for immunity and tissue repair,

vitamin A for vision and skin health, and vitamin E for cellular protection. In addition, fruits

supply B-complex vitamins like B1, B2, B6, and folate, which aid metabolism and blood

formation. Examples include bananas (rich in B6), citrus fruits and guavas (high in vitamin C),

and mangoes and papayas (sources of vitamin A). Consuming a variety of fruits daily promotes

growth, prevents diseases, and supports overall wellbeing. (Ovonramwen et al,. 2024). Fruits

also supply important minerals such as potassium, calcium, magnesium, and iron, which are

vital for muscle function, bone health, and maintaining fluid balance in the body. Furthermore,

they contain phytochemicals and antioxidants like flavonoids and carotenoids that help protect

the body against cell damage and reduce the risk of chronic diseases such as cancer and heart
disease. Overall, fruits are naturally low in fat and protein but rich in beneficial nutrients, making

them an essential part of a balanced diet. Regular consumption of a variety of fruits contributes

to overall wellbeing, vitality, and long-term health. (Ara.,et al 2014)..Therefore, while fruits are

beneficial nutritionally, their physicochemical properties make them highly perishable and

susceptible to microbial contamination.

2.2 Fruits in Health and Nutritional Perspectives

Fruits play a crucial role in promoting human health by supplying essential micronutrients,

dietary fiber, and bioactive compounds that support metabolic and immune functions (World

Health Organization [WHO], 2022). Regular fruit consumption has been associated with reduced

risks of cardiovascular diseases, cancer, and digestive disorders (Okeke et al., 2021).In Nigeria,

fruits such as mangoes, pawpaw, oranges, and watermelon are commonly consumed as snacks or

part of meals due to their availability and affordability (Ameh et al., 2022). They serve as natural

sources of hydration and energy, especially in hot climates like Dutse. Fruits are rich in essential

nutrients and bioactive compounds such as phytochemicals, vitamins, minerals, dietary fiber,

proteins, carbohydrates, carotenoids, and flavonoids, all of which play a vital role in maintaining

overall health. Regular consumption of fresh fruits, nuts, dried fruits, and natural fruit juices

supports the body by supplying necessary nutrients for growth, metabolism, and disease

prevention Including a wide variety of fruits—such as dates, oranges, grapefruits, pomegranates,

cranberries, and nuts—in the daily diet helps manage metabolic disorders like diabetes,

strengthen the immune system, and enhance organ function. Fruits also serve as powerful

antioxidants and anti-inflammatory agents, reducing oxidative stress, improving cardiovascular

health, and lowering the risk of chronic and non-communicable diseases. In essence, fruits are
not just energy-giving foods but natural protectors that sustain physiological balance and

promote long-term wellbeing. However, contamination with pathogenic microorganisms can

offset these benefits, leading to foodborne infections and gastrointestinal illnesses. (Abobatta et

al., [Link], maintaining fruit safety from farm to table is essential for public health.

2.3 Spoilage of Ready-to-Eat Fruits

The spoilage of ready-to-eat (RTE) fruits poses a major concern for food safety, especially when

harmful bacteria such as Listeria monocytogenes are able to grow under refrigeration without

visible signs of deterioration. This psychrotolerant pathogen can multiply even at low

temperatures, increasing health risks before any noticeable spoilage occurs. In some preservation

methods, such as modified atmosphere packaging (MAP)—used to extend shelf life and

maintain freshness—the reduction of normal spoilage microorganisms can unintentionally create

favorable conditions for L. monocytogenes growth. Consequently, fruits may appear fresh and

safe while harboring increased bacterial loads. Conversely, using damaged or low-quality

produce shortens the shelf life, limiting the time available for bacterial proliferation but

compromising overall quality(Cai & Snyder 2019).Thus, spoilage should not always be

considered a reliable indicator of microbial safety in ready-to-eat fruits. Even when visual or

sensory signs of decay are absent, pathogenic microorganisms may already have reached unsafe

levels, emphasizing the need for strict hygienic handling, temperature control, and regular

microbial monitoring throughout processing and storage. Microbial contamination of fruits can

occur at any stage—from cultivation to consumer handling. When fruits are cut to produce fresh-

cut or ready-to-eat forms, their protective skin is broken, exposing nutrient-rich inner tissues that

support microbial growth. Contamination is further increased by poor hygiene among handlers,
the use of contaminated water or utensils, and unsanitary processing environments. Additionally,

open display of fruits in markets exposes them to flies, dust, and airborne pathogens, all of which

contribute to spoilage and potential health risks. (Oyedele et al., 2020).

2.4 Microbial Spoilage Mechanisms in Ready-to-Eat Fruits

Pathogens such as Salmonella and Shiga toxin-producing E. coli can survive for long periods

under refrigeration, posing food safety risks. Factors like poor manure management, climate

changes, and contaminated equipment increase the chances of fruit contamination. Food contact

surfaces and utensils can harbor bacteria, making regular cleaning and disinfection crucial to

prevent biofilm formation and ensure the safety of ready-to-eat fruits. (Kotzekidou et al., 2016).

Fruits and vegetables, like all fresh foods, are highly susceptible to microbial spoilage caused by

fungi, bacteria, yeasts, and molds. Their moist and nutrient-rich nature makes them ideal for

microbial invasion, leading to significant post-harvest losses. Many spoilage problems begin in

the field and are worsened by contaminated wash water, handling equipment, and storage

conditions.

Fungal species such as Alternaria, Botrytis, Penicillium, Fusarium, and Rhizopus, along with

bacterial genera like Erwinia and Pseudomonas, are the main culprits responsible for soft rot and

fruit decay. These infections not only affect fresh produce but also extend to canned and

processed products, sometimes ruining entire batches. Overall, microbial spoilage remains a

major challenge in fruit and vegetable preservation, accounting for a large portion of post-harvest

losses worldwide.( Rawat et al.,2015)

2.5 Factors Determining the Rate of Microbial Growth on Ready-to-Eat Fruits

Intrinsic factors, epiphytic microbiota, structure of product, edible coatings, and packaging

contribute to the safety of ready-to-eat (RTE) foods. Refrigeration throughout the cold-chain is

essential to reduce the growth rate of Listeria monocytogenes. The survival of Salmonella, and

Shiga-toxin-producing Escherichia coli for extended periods in chilled conditions are of concern.

Climate changes affecting ambient temperature, increased use of improperly composted manure,

and extreme weather conditions influence contamination and internalization of enteric pathogens

in fresh produce. Food preparation surfaces, slicing equipment, and dishcloths used for cleaning

utensils, may be reservoirs for bacterial contamination of RTE foods. (Kotzekidou et al., 2016).

Warm temperatures and high humidity provide ideal conditions for the growth of

microorganisms that cause post-harvest decay in fruits and vegetables. The type of spoilage

organism often depends on the acidity of the produce—fungi tend to thrive in more acidic

tissues, while bacteria commonly affect produce with a pH above 4.5. For instance, bacterial

soft rot in potatoes caused by Ceratocystis fimbriata and watery soft rot in carrots caused by

Sclerotinia sclerotiorum are typical examples. Overall, improper temperature and humidity

control significantly accelerate microbial deterioration after harvest.( Rawat et al.,2015) Both

intrinsic and extrinsic factors play crucial roles in determining the microbial safety and shelf life

of foods. Intrinsic factors are the natural characteristics of the food—such as nutrient

composition, pH, moisture content (water activity), oxidation-reduction potential, and natural

antimicrobial compounds—that influence microbial growth. Extrinsic factors, on the other

hand, relate to external environmental conditions like temperature, humidity, surrounding gases,

and the presence of other microorganisms. Even small changes in these conditions can affect

food stability and encourage the growth of spoilage or pathogenic microbes. Since different

microorganisms respond differently to these factors, maintaining food safety requires a balanced,
multi-factor approach that considers both internal and external influences on microbial activity.

( Rolfe et al.,2020).

2.8 Some Bacteria Associated with the Spoilage of Ready-to-Eat Fruits

Foodborne disease outbreaks result from various microbial contaminants, including bacteria,

viruses, fungi, parasites, and their toxins. Among these, bacterial pathogens such as Listeria

monocytogenes, Escherichia coli, and Salmonella enterica are leading causes of contamination

linked to fruits and vegetables. Between 2004 and 2012, these bacteria were responsible for

about 42% and 36% of produce-related outbreaks in the European Union and the United States,

respectively. Other significant bacterial agents associated with foodborne illnesses include

Clostridium, Shigella, Campylobacter, Vibrio cholerae, and Bacillus cereus. Collectively, these

pathogens highlight the critical need for proper hygiene, storage, and handling practices to

ensure the microbial safety of fresh and ready-to-eat foods. (Abdelshafy et al., 2025).

2.8.1 [Link]

Escherichia coli is commonly introduced into fruits through fecal contamination of irrigation

water, soil, or improper handling. Pathogenic strains, such as E. coli O157:H7, can persist on

fruit surfaces and may lead to serious foodborne infections when hygiene standards are poor

(Alegbeleye, Singleton, & Sant’Ana, 2018). Similarly, Staphylococcus aureus contamination

often occurs through human contact during processing and retail display. This bacterium
produces enterotoxins that can cause food poisoning if fruits are stored under warm or humid

conditions (Abadias et al., 2008).

2.8.2 Salmonella enterica

Salmonella enterica is another well-documented pathogen associated with RTE fruits.

Contamination may occur from irrigation water, animal manure, or unhygienic post-harvest

handling. It has been linked to several outbreaks involving fresh produce such as melons and

tomatoes (Olaimat & Holley, 2012). Listeria monocytogenes, on the other hand, is notable for its

psychrotolerant nature, allowing it to grow even under refrigeration. This organism can form

biofilms on fruit surfaces or processing equipment, posing a serious risk especially in pre-cut

fruit salads (Carpentier & Cerf, 2011).In addition to pathogenic bacteria, spoilage species such as

Pseudomonas fluorescens contribute significantly to quality deterioration. This psychrotrophic

bacterium thrives in chilled storage environments, leading to soft rot, slime formation, and

unpleasant odors in stored fruits (Liu et al., 2019).

2.8.3 Staphylococcus aureus

Staphylococcus aureus is one of the most common bacterial contaminants associated with the

spoilage and potential health risks of ready-to-eat (RTE) fruits. It is a Gram-positive,

facultative anaerobic bacterium frequently found on the skin, nasal passages, and hands of

humans, making human handling the primary source of contamination during fruit
processing, packaging, and retail display (Abadias et al., 2008; Kadariya, Smith, & Thapaliya,

2014).When RTE fruits such as sliced melons, apples, and pineapples are exposed to improper

hygienic practices or temperature abuse, S. aureus can multiply rapidly. This bacterium produces

heat-stable enterotoxins, which remain active even after mild heat treatment or refrigeration,

causing food poisoning characterized by nausea, vomiting, and abdominal cramps (Le Loir,

Baron, & Gautier, 2003).

2.9 Control Measures

The global demand for fresh and ready-to-eat (RTE) produce has risen, but so has the incidence

of foodborne illnesses linked to contaminated(RTE)fruits. Since fresh and minimally processed

vegetables are not treated in ways that eliminate human pathogens, various handling steps—such

as selection, washing, peeling, cutting, and packaging—can promote contamination or cross-

contamination. To reduce these risks, producers are advised to follow the Codex Code of

Hygienic Practice for Fresh Fruits and Vegetables and implement control measures. Applying

mild preservation or disinfection methods, along with traditional chemical or physical

treatments, can help improve the microbial safety of these products. (Castro-Ibáñez & Allende

2017). The rising consumption of ready-to-eat fruits in developing countries, driven by changing

lifestyles, has been accompanied by growing food safety [Link] fruits are often

contaminated with pathogenic microbes such as Escherichia coli, Listeria monocytogenes,

coliforms, and Salmonella, posing health risks to consumers. To mitigate these threats and

extend shelf life, various control measures—including refrigeration, modified atmosphere

packaging, and emerging preservation technologies—are being employed to maintain the

microbiological safety and quality of such products. (Mir & Roohinejad 2018).
 CHAPTER THREE

3.0 METHODOLOGY

3.1 STUDY AREA.

The study will take place in Dutse metropolis, the administrative center of Jigawa State,

located in Nigeria’s northwest. Home to over 430,000 residents, Dutse is a rapidly

developing urban hub with a large presence of street food vendors. The city lies within a

tropical savanna zone, experiencing extreme seasonal shifts—ranging from intensely hot dry

periods to humid, rainy months, with temperatures typically between 12°C and 40°C

(Wikipedia, 2025).Although official statistics indicate substantial access to clean water,

frequent shortages remain a persistent challenge, compelling many households and vendors

to depend on private water suppliers .While Jigawa has been recognized as Nigeria’s first

Open Defecation Free (ODF) state, the risk of environmental contamination—especially in

informal food settings—continues to pose public health concerns (UNICEF, 2023;

Vanguard, 2023).

3.2 sample collection.

A total of 18 samples,6 samples each of watermelon (citrus vulgari), pineapple (Ananas

comosus) and pawpaw (Asimina triloba), will be purchased randomly from Dutse ultra-

modern, old markets and Hakimi street. Fruit samples will be collected aseptically into

sterile polythene bags, stored in cool boxes, and processed for microbial analysis within 24

hours, following standard food microbiology protocols (Igbeneghu & Abdu, 2014).And

transported to Microbiology laboratory of the Department of Microbiology and

biotechnology Federal University Dutse Jigawa State.

3.3 Sample preparation.

One hundred millilitres (100 mL) of sterile distilled water is used to wash twenty-five grams

(25 g) of each fruit sample. The rinse water is collected and subjected to ten-fold serial

dilution as described by (Cheesbrough et al,.2006). From the 10⁻¹, 10⁻², and 10⁻³ dilutions,

0.1 mL aliquots are pour-plated onto three types of media: Nutrient Agar (NA) for total

aerobic bacterial count, MacConkey Agar for Escherichia coli, and Mannitol Salt Agar

(MSA) for Staphylococcus aureus.( Yuguda et al,.2025). All plates are incubated at 37 °C

for 24 hours, after which colony characteristics are observed. Presumptive [Link] and

[Link] colonies are further subjected to biochemical confirmation tests.

3.3.1 Serial dilution.

A tenfold serial dilution is carried out to reduce the microbial concentration for enumeration.

Initially, 25 grams of the fruit sample are homogenized in 225 mL of sterile buffered

peptone water to obtain a 10⁻¹ dilution. Subsequently, 1 mL of this homogenate is

transferred into 9 mL of sterile diluent to prepare the 10 ⁻² dilution. This process is repeated

stepwise up to the required dilution level (e.g., 10⁻⁶), using fresh sterile pipette tips at each

step to avoid cross-contamination. Each dilution is thoroughly mixed manually by gentle

shaking before proceeding to the next. The resulting dilutions are used for plating on

selective media to isolate and enumerate [Link] and [Link].( Cheesbrough et al,.2006).

3.3.2 Media preparation.

Nutrient agar is prepared by dissolving 28 grams of nutrient agar powder in 1 liter of

distilled water. The mixture is heated gently while stirring until complete dissolution of the

agar powder occurs. The medium is then sterilized by autoclaving at 121°C and 15 minutes.

After sterilization, the medium is allowed to cool to approximately 45°C before being

poured aseptically into sterile Petri dishes. The plates are left to solidify at room temperature
and stored inverted at 4°C until use. To prepare Mannitol Salt Agar, weigh 55 grams of

MSA powder and dissolve it in 1 liter of distilled water. Heat the solution while stirring to

completely dissolve the medium. Sterilize by autoclaving at 121°C for 15 minutes. After

sterilization, cool the medium to approximately 45°C. Pour the molten agar aseptically into

sterile Petri dishes and allow it to solidify at room temperature. Once solidified, invert the

plates and store them at 4°C until use. To prepare EMB agar, weigh 39 grams of EMB agar

powder and dissolve it in 1 liter of distilled water. Heat the solution while stirring

continuously until the powder is completely dissolved. Sterilize the medium by autoclaving

at 121°C and 15 minutes. After autoclaving, cool the medium to approximately 45°C before

pouring aseptically into sterile Petri dishes. Allow the agar to solidify at room temperature

and store the plates inverted at 4°C until use. EMB agar is a selective and differential

medium used primarily for the isolation of gram-negative bacteria, especially Escherichia

coli, which produces colonies with a characteristic metallic green sheen. Mannitol Salt Agar

is a selective and differential medium used primarily for the isolation of Staphylococcus

aureus. The high salt concentration (7.5% NaCl) inhibits most bacteria other than

staphylococci, while mannitol fermentation leads to acid production, causing the phenol red

indicator to turn yellow around fermenting colonies. (Cheesbrough et al,.2006).

3.3.3 Sample inoculation and incubation.

Aliquots (0.1 mL) from appropriate serial dilutions are aseptically inoculated onto EMB agar

for E. coli and MSA for S. aureus using the spread plate method. Plates are briefly dried and

incubated inverted at 37°C for 24 [Link] individual colonies will be counted using

colony counter for each plate (Cheesbrough et al,. 2006).After incubation, colonies showing
characteristic features metallic green sheen for [Link] and yellow with a yellow zone for

[Link] are identified and confirmed by biochemical tests.

3.3.4 Gram staining.

Gram staining is used to confirm the identity of isolates based on cell wall structure and

morphology. A bacterial smear is prepared, air-dried, and heat-fixed, then sequentially

stained with crystal violet, iodine, alcohol (for decolorization), and safranin. Staphylococcus

aureus appears as Gram-positive cocci (purple), while Escherichia coli appears as Gram-

negative rods (pink). This step helps verify results from selective media before further

biochemical testing. ([Link] al,. 2006).

3.3.5 Biochemical tests

Biochemical tests will be used to confirm the identity of E. coli and S. aureus after isolation.

[Link] is confirmed using IMViC tests and oxidase tests—typically indole and methyl red

positive, citrate and oxidase negative. S. aureus is identified by positive catalase and

coagulase reactions. These tests are sufficient to verify isolates from selective media.

3.3.6 Indole Test

The indole test will be carried out by inoculating the isolate into tryptone broth and

incubating at 37 °C for 24 hours. After incubation, 0.5 mL of Kovács reagent will be added.

A red or pink ring at the surface will be recorded as indole positive, which is characteristic

of E. coli (Cheesbrough et al., 2006).

3.3.7 Methyl Red and Voges–Proskauer Tests

MR-VP broth will be inoculated with the test organism and incubated at 37 °C for 24 hours.

For the MR test, five drops of methyl red indicator will be added to an aliquot of the broth; a

persistent red color indicates MR positive. For the VP test, reagents α-naphthol (VP A) and

40% KOH (VP B) will be added; a red color within 30 minutes will be read as VP positive.

E. coli is typically MR positive and VP negative ([Link] al,. 2006).

3.3.8 Oxidase Test

Fresh colonies will be rubbed onto filter paper impregnated with oxidase reagent

(tetramethyl-p-phenylenediamine dihydrochloride). Development of a dark purple color

within 30 seconds will indicate an oxidase positive reaction. E. coli is oxidase negative

(ASM. 2023).

3.3.9 Catalase Test

A portion of the isolate will be placed on a clean glass slide and one drop of 3% hydrogen

peroxide will be added. Immediate bubble formation will indicate catalase positive,

distinguishing staphylococci (positive) from streptococci (negative) (Cheesbrough et al.,

2006).

3.4.0 Coagulase Test

The slide coagulase test will be performed as a rapid screening method by mixing colonies

with plasma and observing for clumping within 15 seconds. The tube coagulase test will be

used for confirmation, where rabbit plasma inoculated with the isolate will be incubated at

37 °C and checked for clot formation at 1 hour and again at 24 hours. A clot indicates

coagulase positive, confirming S. aureus (Cheesbrough et al., 2006).

3.4.1 Bacterial Data

Data Analysis Data from the 18 fruit samples will be analyzed using descriptive statistics.

Mean, minimum, and maximum CFU/g values will be calculated for each fruit type.

Prevalence of E. coli and S. aureus will be expressed as frequencies and percentages. Results

will be presented in tables, with bar charts showing prevalence across fruit types and pie

charts showing the overall proportion of positive and negative samples (Cheesbrough et al.,

2006).

3.4.2 Questionnaire Data Analysis

Data from the vendor questionnaires will be coded and analyzed using descriptive statistics.

Frequencies and percentages will be calculated for categorical variables such as sex,

education level, washing practices, and hygiene behaviors. Likert-scale responses will be

summarized using means and standard deviations. The questionnaire results will be used to

assess possible associations between vendor practices and bacterial contamination levels in

the fruit samples (Field et al., 2013).

 CHAPTER ROUR

4.0 Results

4.1 Samples Distributions

A total of 18 ready-to-eat fruit samples were collected from three locations in Dutse metropolis:

Old Market, New Market, and Hakimi Street. Each location supplied two (2) samples of

watermelon, pineapple, and pawpaw, giving six (6) samples per location and eighteen (18)

samples in total.

Table 4.1: Samples Distributions from Dutse Modern Market, Old Market and Hakimi

Street

Location Watermelo Pineapple Pawpaw Total

Old market 2 2 2 6

New market 2 2 2 6

Hakimi 2 2 2 6

street

Total 6 6 6 18
The table confirms equal distribution of samples across fruits and locations. This uniformity

allows valid comparison of microbial contamination levels.

4.2 Total Bacterial Count (CFU/mL)

All samples were cultured on Nutrient Agar using dilution factor 10 ⁻⁶ and 0.1 mL plating

volume. CFU was calculated using: CFU/ml = Colonies × 10^7.

Table 4.2: Bacterial load of ready to eat fruits collected from Dutse modern market, Old market

and Hakimi street

Sample Location Fruit Colony Dilution Volume CFU/mL

Count (mL)
Code

OM-W1 Old Watermelon 58 10⁻⁶ 0.1 5.8 × 10⁸

Market

OM-W2 Old Watermelon 83 10⁻⁶ 0.1 8.3 × 10⁸

Market

OM-P1 Old Pineapple 92 10⁻⁶ 0.1 9.2 × 10⁸

Market

OM-P2 Old Pineapple 93 10⁻⁶ 0.1 9.3 × 10⁸

Market

OM-PA1 Old Pawpaw 115 10⁻⁶ 0.1 1.15 × 10⁹

Market

OM-PA2 Old Pawpaw 210 10⁻⁶ 0.1 2.1 × 10⁹

Market

NM-W1 New Watermelon 165 10⁻⁶ 0.1 1.65 × 10⁹

Market

NM-W2 New Watermelon 115 10⁻⁶ 0.1 1.15 × 10⁹

 Market

NM-P1 New Pineapple 97 10⁻⁶ 0.1 9.7 × 10⁸

Market

NM-P2 New Pineapple 87 10⁻⁶ 0.1 8.7 × 10⁸

Market

NM-PA1 New Pawpaw 159 10⁻⁶ 0.1 1.59 × 10⁹

Market

NM-PA2 New Pawpaw 42 10⁻⁶ 0.1 4.2 × 10⁸

Market

HS-W1 Hakimi Watermelon 27 10⁻⁶ 0.1 2.7 × 10⁸

Street

HS-W2 Hakimi Watermelon 170 10⁻⁶ 0.1 1.7 × 10⁹

Street

HS-P1 Hakimi Pineapple 30 10⁻⁶ 0.1 3.0 × 10⁸

Street

HS-P2 Hakimi Pineapple 115 10⁻⁶ 0.1 1.15 × 10⁹

Street

HS-PA1 Hakimi Pawpaw 156 10⁻⁶ 0.1 1.56 × 10⁹

Street

HS-PA2 Hakimi Pawpaw 38 10⁻⁶ 0.1 3.8 × 10⁸

 Street

Key: OLM=Old market, NM=New market, HS=Hakimi street, W=Watermelon, PA=Pineapple,

P=Pawpaw

Bacterial load varied across samples. The highest CFU/mL was recorded in pawpaw from Old

Market (2.1 × 10⁹), while watermelon from Hakimi Street recorded the lowest (2.7 × 10⁸).

Table 4.3: Isolation on EMB and MSA Between the samples.

E. coli was most frequent in watermelon, while pineapple had the lowest detection rate. High

EMB positivity indicates fecal contamination through water, handling, or knives.

Fruit No. of Samples EMB Positive (E. MSA Positive (S.

coli) aureus)

Watermelon 6 5 4

Pineapple 6 3 3

Pawpaw 6 4 3

Total 18 12 10

Table 4.4: Distributions of Bacteria on EMB and MSA Plates among locations

A higher number of EMB-positive samples were recorded in Hakimi Street and New Market,

indicating heavy contamination by lactose-fermenting coliforms such as E. coli. The presence of

MSA-positive isolates also reveals possible contamination with Staphylococcus aureus,

especially in street-vended fruits.

Sample Source Number of EMB Positive (E. MSA Positive (S.

Samples coli) aureus)

Old Market 6 4 (66.7%) 3 (50%)

Hakimi Street 6 6 (100%) 5 (83.3%)

New Market 6 5 (83.3%) 5 (83.3%)

4.5 Morphological and Biochemical tests of the Organisms

E. coli isolates were confirmed by metallic green sheen colonies on EMB agar and positive

indole test, while citrate test was negative. S. aureus isolates were identified by yellow colonies

on MSA, catalase positivity, and mannitol fermentation.

Table 4.5: Culture, Morphological and Biochemical Characteristics of isolates

Cultural Cat. Coa. Ind. Mr. Cit. Vp. Isolates.

characterization

Round, golden + + _ _ _ _ S.

yellow colonies Aureus

on MSA

Round, Green _ + + + _ _ E. coli

Metallic sheen on

EMB

Key:cat=Catalase test, Coa=coagulase test, Ind=indole test, Cit=citrate utilization test,

MR=methyl red, VP=voges proskaure, MSA=Manitol salt agar, EMB=, -=Negative, + =Positive

4.6 Frequency Distribution of Total Bacterial Count.

The frequency distribution shows that most fruit samples (27.78%) had bacterial counts between

81–120 ×10^7 CFU, indicating moderate contamination. Only 5.56% of samples recorded

extremely high bacterial load above 200 ×10^7 CFU. About 33.3% of samples fell below 80

×10^7 CFU, while 33.3% were above 120 ×10^7 CFU. The wide variation in bacterial load

suggests differences in vendor hygiene, handling, and environmental exposure among sampling

locations.

Table 4.6 Frequency Distribution of Total Bacterial Count

Class Interval (CFU Frequency Percentage (%)

×10^7)

0 – 40 3 16.67

41 – 80 3 16.67

81 – 120 5 27.78

121 – 160 3 16.67

161 – 200 3 16.67

201 – 240 1 5.56

Total 18 100

Table 4.7: Frequency Distribution of Isolates

 Bacterial Isolate Frequency Percentage (%)

E. coli positive 12 66.7%

S. aureus positive 10 55.6%

Total Positive Samples 18 100%

 Detection of Counts per locations
6.5 6
5.5 5 5 5
4.5
No. of positive samples

4
3.5 3
2.5

1.5

0.5
HAKIMI NEW-M OLD-M
E. coli 6 5 4
S. Aureus 5 5 3

E. coli S. Aureus

Detection counts Per Fruits

5.5 5

4.5 4 4
Number of positive samples

3.5 3 3 3

2.5

1.5

0.5
Watermelon Pineapple Pawpaw

[Link] 5 3 4 NaN
[Link] 4 3 3 NaN
Column1 NaN NaN NaN NaN

[Link] [Link]
Percentage of Positive Isolates

55.60%

66.70%

[Link] positive [Link] positive

4.8: Questionnaire Analysis

Vendors frequently handled money alongside fruits and displayed fruits uncovered, exposing

them to flies, dust, and human contact. These practices correlate strongly with high

contamination.

Table 4.9:

Variable Yes (%) No (%)

Wash fruits before selling 40 60

Display fruits uncovered 78 22

Handle money and fruits 89 11

together

Sell near drainage/waste 44 56

Use clean water source 55 45

 CHAPTER FIVE

5.0 Discussion

The findings of this study reveal that ready-to-eat (RTE) fruits sold by street vendors in Dutse

metropolis harbor significant levels of bacterial contamination. The presence of Escherichia coli

and Staphylococcus aureus in some fruit samples indicates poor hygienic handling, use of

contaminated water, and exposure to environmental pollutants. These results agree with earlier

studies by (Mensah et al.2002), who reported similar contamination patterns in open-market

foods. The bacterial load across vending locations varied, with higher counts in areas

characterized by heavy human traffic, dust, and poor sanitation. Fruits sold along major

roadsides recorded the highest microbial levels, possibly due to constant exposure to dust and

vehicle emissions. The detection of E. coli suggests fecal contamination, either from dirty water

or poor personal hygiene among vendors. Staphylococcus aureus, which is commonly found on

human skin, points to direct contamination during peeling, slicing, or packaging. These findings

demonstrate that although RTE fruits are nutritious and widely consumed, they can be potential

vehicles for foodborne infections when proper hygiene practices are not observed.

5.1 Conclusion

This study concludes that ready-to-eat fruits sold by street vendors in Dutse metropolis are

susceptible to microbial contamination, with some samples testing positive for Escherichia coli

and Staphylococcus aureus. Poor hygienic conditions, inadequate washing, and environmental

exposure contribute significantly to bacterial load. The findings highlight the need for strict

hygiene practices and regulatory oversight to protect public health.

5.2 Recommendations
1. Vendors should always wash fruits with clean, potable water before cutting and serving.

2. Fruit handlers must maintain proper personal hygiene, including handwashing and use of clean

utensils.

[Link] authorities should create policies to regulate street vending and enforce sanitation

standards.

4. Public health education campaigns should be conducted to create awareness on food safety.

5. Consumers should purchase RTE fruits from vendors who practice visible hygiene and proper

food handling.
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