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MICROBIOLOGICAL ANALYSIS OF STREET FOOD IN NAIROBI

EVANS JEREMIAH OMBUI


I23/5486/2020
BSC MICROBIOLOGY AND BIOTECHNOLOGY

A PROJECT REPORT SUBMITTED IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR


COURSE UNIT SBT 414 RESEARCH PROJECT IN THE BSC. (MICROBIOLOGY AND
BIOTECHNOLOGY) DEGREE PROGRAM

DEPARTMENT OF BIOLOGY
FACULTY OF SCIENCE AND TECHNOLOGY
UNIVERSITY OF NAIROBI

April, 2024

i
DECLARATION

Student’s Declaration
I hereby declare that this research project report is my original work and has not been submitted
elsewhere, either in part or in full, for the award of either a degree or a diploma in the University of
Nairobi or any other institution.

Student’s name. Evans Jeremiah Ombui


Reg. No. I23/5486/2020

Sign: ……………. Date: ………………

Declaration by Project Supervisor


This project report has been submitted for examination with my approval as the University of Nairobi
supervisor.

Supervisor’s Name: Prof. Sheila Okoth

Sign: ……………. Date: ………………...

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ACKNOWLEDGEMENT

I would like to express my sincere gratitude to several individuals and organizations for supporting me
throughout my Graduate study. Firstly, glory be to God for all was made possible by Him, He gave me
grace and strength to complete this study amidst various challenges. I wish to express my sincere gratitude
to my supervisor, Professor Sheila Okoth, for her enthusiasm, patience, insightful, comments, helpful
information, practical advice and unceasing ideas that have helped me tremendously at all times in my
research and writing of this thesis. Her knowledge, profound experience and professional expertise in
Mycology has enabled me to complete this research successfully. Without her support and guidance, this
project would not have been possible. I could not have imagined having a better supervisor in my study.
I also wish to express my sincere thanks to the University of Nairobi for accepting me into the graduate
program.

I would also like to thank the University of Nairobi Mycology laboratory technician Madam Gertrude
Shikote and Bacteriology laboratory technician Mr. Wachira for assisting me by supplying the equipment
that was essential and vital, without which I would not have been able to perform efficiently on this project.
In addition, I am deeply indebted to the Ministry of Education, University of Nairobi work-study program
for granting me the financial support that enabled me to complete my undergraduate studies successfully.
I’d also like to thank my friends and parents for their support and encouragement as I worked on this
assignment.

iii
TABLE OF CONTENTS
ACKNOWLEDGEMENT ....................................................................................................................... iii

Lists of Figures ......................................................................................................................................... vi


List of Tables ........................................................................................................................................... vii
Abbreviations and Acronyms ............................................................................................................... viii
ABSTRACT .............................................................................................................................................. ix
CHAPTER ONE: INTRODUCTION ..................................................................................................... 1
1.1 Background Information ................................................................................................................... 1
1.2 Problem Statement ............................................................................................................................ 2
1.3 Project Justification ........................................................................................................................... 3
1.4 Objectives .......................................................................................................................................... 3
1.4.1 Broad Objective .......................................................................................................................... 3
1.4.2 Specific Objective....................................................................................................................... 3
CHAPTER TWO: LITERATURE REVIEW ........................................................................................ 4
2.1 Overview of Street Foods.................................................................................................................. 4
2.1.1 Background of Street Foods ....................................................................................................... 4
2.1.2 Significance of Street Foods in Urban Settings .......................................................................... 5
2.1.3 Emergence of Concerns on Safety and Hygiene ........................................................................ 5
2.2 Microbial Contamination in Street Foods ......................................................................................... 6
2.2.1 Bacteria ....................................................................................................................................... 6
2.2.2 Fungi and Molds ......................................................................................................................... 7
2.2.3 Viruses ........................................................................................................................................ 7
2.2.4 Parasites ...................................................................................................................................... 7
2.3 Factors Contributing to Microbial Contamination ............................................................................ 8
2.3.1 Poor Personal Hygiene Practices ................................................................................................ 8
2.3.2 Water Quality and Environmental Factors ................................................................................. 8
2.3.3 Equipment and Packaging Concerns .......................................................................................... 9
2.3.4 Lack of Training and Awareness ................................................................................................ 9
2.4 Microbial indicators of food quality and safety ................................................................................ 9
2.4.1 Total Viable Count (TVC) ........................................................................................................ 10
2.4.2 Coliforms .................................................................................................................................. 10
2.4.3 Standard Plate Count (SPC) ..................................................................................................... 10

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2.4.4 Molds and Yeasts...................................................................................................................... 11
2.5 Hygiene and Regulatory Challenges ............................................................................................... 11
2.6 Global Perspectives on Street Food Safety ..................................................................................... 11
MATERIALS AND METHODS ........................................................................................................... 13
3.1 Study Area ....................................................................................................................................... 14
3.2 Sample Collection ........................................................................................................................... 14
3.3 Media Preparation ........................................................................................................................... 15
3.3.1 Nutrient Agar ............................................................................................................................ 15
3.3.2 Potato Dextrose Agar................................................................................................................ 15
3.4 Isolation of Microorganisms ........................................................................................................... 16
3.4.1 Serial Dilutions ......................................................................................................................... 16
3.4.2. Isolation of Bacteria on Nutrient Agar .................................................................................... 16
3.4.3 Isolation of Fungi on the PDA .................................................................................................. 16
3.4.5 Sub-culturing ............................................................................................................................ 17
3.5 Characterization of Isolates ............................................................................................................. 17
3.5.1 Colony Morphology Characteristics ......................................................................................... 17
3.5.2 Gram Staining ........................................................................................................................... 17
CHAPTER FOUR: RESULTS .............................................................................................................. 18
4.1 Bacterial Analysis ............................................................................................................................... 18
4.1.1 Isolated Bacteria on Nutrient Agar ........................................................................................... 18
4.1.2 Subculture bacteria ................................................................................................................... 19
4.1.3 Morphological Characterization of the Isolates........................................................................ 19
4.1.4 Gram Stain ................................................................................................................................ 20
4.2 Fungal Analysis ............................................................................................................................... 20
4.2.1 Isolated Fungi on PDA ............................................................................................................. 20
4.2.2 Characterization of Isolated Fungi ........................................................................................... 22
4.2.3 Microscopic Characterization of Isolated Fungi ...................................................................... 24
4.2.4 Morphological Characterization of the Isolated Fungi ............................................................. 25
CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATIONS ....................... 26
5.1 Discussion ....................................................................................................................................... 26
5.2 Conclusion....................................................................................................................................... 27
5.3 Recommendations ........................................................................................................................... 28

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Lists of Figures
Figure 1: Location of the study area ........................................................................................................ 14
Figure 2: Samples collected for the study ................................................................................................ 15
Figure 3: Isolated bacteria from fish ........................................................................................................ 18
Figure 4: Isolated bacteria from sausage ................................................................................................. 18
Figure 5: Isolated bacteria from banana .................................................................................................. 18
Figure 6: Isolated bacteria from Roasted maize ...................................................................................... 18
Figure 7: Subculture of bacteria isolated from fish ................................................................................. 19
Figure 8: Subculture of bacteria isolated from Sausage .......................................................................... 19
Figure 9: Subculture of bacteria isolated from Roasted maize ................................................................ 19
Figure 10: Subculture of bacteria isolated from Banana ......................................................................... 19
Figure 11: Bacillus sp. ............................................................................................................................. 20
Figure 12: Streptococcus sp. .................................................................................................................... 20
Figure 13: E. coli ..................................................................................................................................... 20
Figure 14: Fungi isolated from Roasted Maize ....................................................................................... 21
Figure 15: Fungi isolated from Sausage .................................................................................................. 21
Figure 16: Fungi isolated from Banana ................................................................................................... 21
Figure 17: Fungi isolated from Fish ........................................................................................................ 22
Figure 18: Sub-culture of Fungi Isolated from Fried Fish ....................................................................... 22
Figure 19: Sub-culture of Fungi Isolated from Sausage .......................................................................... 22
Figure 20: Sub-culture of Fungi Isolated from Roasted Maize ............................................................... 23
Figure 21: Sub-culture of Fungi Isolated from Banana ........................................................................... 23
Figure 22: Fusarium sp. Isolated from Fish ............................................................................................ 24
Figure 23: Trichophyton tonsurans sp. Isolated from sausage ................................................................ 24
Figure 24: Fusarium sp. Isolated from Roasted Maize ........................................................................... 24
Figure 25: Fusarium sp. Isolated from Banana ....................................................................................... 24

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List of Tables
Table 1: Morphological characteristics of Bacterial isolates. ................................................................................. 20
Table 2: Table 1: Morphological characteristics of Fungal isolates. ..................................................................... 25

vii
Abbreviations and Acronyms

SDA– Sabouraud Dextrose Agar


PDA– Potato Dextrose Agar
CBD– Central Business District
UON– University Of Nairobi
P.m– Post Meridiem
Sp– species
ETEC– Enterotoxigenic Escherichia coli
EHEC– Enterohemorrhagic Escherichia coli
E.coli– Escherichia coli
cfu/g– colony forming unit per gram
Prof.– professor
Reg. No– Registration number
Et al – et alia
TVC– Total Viable Count
SPC– Standard Plate Count

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ABSTRACT

Street foods represent a significant component of the culinary landscape in Nairobi, offering convenience
and diversity to urban consumers. However, concerns regarding the microbiological and mycological
safety of these foods persist. This study aimed to assess the microbial and fungal quality of selected street
foods in Nairobi, focusing on banana, sausage, fried fish, and roasted maize.

Food samples were collected randomly from the City Market street in Nairobi.Using laboratory methods
of isolation of microbial communities, Bacillus species, Enterococci species, Staphylococcus aureus,
Staphylococcal enterotoxins and Aspergillus species, Fusarium species, and penicillium species were
isolated from the food samples. The presence of microorganisms in street foods suggests hygiene practices
should be improved.

microbiological analysis was conducted to enumerate indicator organisms, including total coliforms,
Escherichia coli, Staphylococcus aureus, and Streptococcus., measured in colony-forming units (CFU)
per gram, mycological analysis was performed to identify fungal species present in the samples.

The results revealed microbial contamination in all food items examined. Total coliform counts ranged
from <10 to 10^5 CFU/g, with the highest counts observed in fried fish samples.

Mycological analysis revealed fungal contamination in all food items. The predominant fungal species
identified included Aspergillus spp., Trichophyton app., Penicillium spp., and Fusarium spp.

The presence of pathogenic bacteria and potentially toxigenic fungi in street foods poses a significant
public health risk to consumers. Factors contributing to contamination include poor hygiene practices
during food preparation, inadequate storage conditions, and environmental contamination.

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CHAPTER ONE: INTRODUCTION
1.1 Background Information
Street food plays an important role in the daily diet of many people living in Nairobi, Kenya (Kariuki,
2018). Popular street food includes mandazi, a sweet and fried pastry, nyama choma: grilled spiced
meat. Mutura, is a spicy sausage made from ground meat and spices. Githeri, a hearty mixture of cooked
corn and beans; Viazi Karai (Chips), seasoned fries. Mahindi Choma (Roasted Maize), roasted corn with
spices. Kebabs, skewered, and grilled meat. Chapati, a soft unleavened flatbread. Samosa, triangular
pastry stuffed with spices. Roasted peanuts and corn offer locals and tourists a variety of delicious and
affordable options. These street foods contribute to Nairobi's dynamic and rich culinary offering.

Contribute to local food culture and provide convenient and affordable options for a diverse population.
However, concerns about the safety and hygiene of street food have become a significant public health
issue (Rakha et al., 2022). The unregulated sale of street foods, coupled with poor food handling
practices and lack of hygiene infrastructure, has raised questions about the microbial quality of these
foods and their potential impact on consumer health (Muhonja & Kimathi, 2014).

Nairobi is a bustling city center with a variety of street vendors dotted in different districts. These
providers cater to a wide range of consumers, including office workers, students, and residents, and are
an integral part of local food culture. Despite its popularity, the potential health risks associated with
street food cannot be ignored. Factors such as improper food handling, unhygienic cooking
environments, and improper storage conditions can contribute to microbial contamination, which can
pose a risk of foodborne illness to consumers (Birgen et al., 2020).

Street food is susceptible to many types of microorganisms, including bacteria, viruses, parasites, fungi,
and molds (Aytac & Taban, 2014). The microbial composition of street food depends on factors such as
food handling, storage conditions, and the environment in which the food is prepared. Bacterial
contaminants commonly include Escherichia coli (E. coli), which is associated with fecal contamination
and gastrointestinal infections, Salmonella, which is known to cause food poisoning, and
Staphylococcus aureus, which produces toxins that cause food poisoning (Balali et al., 2020). Viruses
such as norovirus, which is highly contagious and causes gastroenteritis, and hepatitis A virus, which is
transmitted through contaminated food, pose additional health risks.

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Parasites such as Giardia lamblia and Cryptosporidium can cause gastrointestinal disease. Fungi such as
Aspergillus spp. They can produce mycotoxins and contaminate stored grains, nuts, and dried fruits.
Certain molds also occur, including Penicillium can produce mycotoxins under certain conditions.
Understanding the presence and distribution of these microorganisms is important for assessing the
health risks associated with street food consumption (Kamala & Kumar, 2018). Hygiene practices,
proper food handling, and effective hygiene measures are important to minimize the risk of microbial
contamination and protect public health. Regular monitoring and analysis play an important role in
ensuring the safety of street food consumption in various urban environments (Ferrari et al., 2021).

In Nairobi, where a large proportion of the population relies on street food for their daily diet,
understanding the microbial quality of these foods is essential for public health management. Despite the
potential risks, there are clear gaps in the scientific literature regarding the specific types of
microorganisms present in Nairobi Street food, their prevalence, and suppliers' compliance with
sanitation and hygiene standards (Kariuki, 2018).

This research project aims to fill this gap by conducting a detailed microbiological analysis of street
food in Nairobi. The results not only contribute to the scientific understanding of the microbial quality
of street foods, but also inform policymakers, regulators, and others about the measures needed to
improve food safety and protect consumer health. The ultimate goal is to establish evidence-based
interventions to reduce the risks associated with microbial contamination of street food, thereby ensuring
a safer and healthier urban food environment in Nairobi, Kenya.

1.2 Problem statement


Street food consumption is widespread in Nairobi, Kenya, and concerns about the safety and hygiene of
these widely consumed foods pose a potential public health risk. Unregulated street food marketing
practices and poor adherence to proper food handling and hygiene procedures have raised concerns
about the prevalence of microbial contamination in these foods. The lack of comprehensive research on
the nature and extent of microbial contamination in street food in Nairobi leaves significant gaps in our
understanding of the associated health risks. Therefore, there is a need to investigate and evaluate the
microbial quality of street food, identify specific pathogens, and evaluate sellers' compliance with
hygiene standards. Addressing this gap will improve food security, minimize health risks, and contribute
to the overall well-being of the diverse population that relies on street food in Nairobi.

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1.3 Project Justification
This research project on the microbial analysis of street food is of great importance because these foods
are widely consumed and concerns about their safety and hygiene are important. The fact that street food
sales are unregulated, coupled with the lack of compliance with food handling and hygiene standards,
emphasizes the need for this research. With lack of comprehensive research specific to Nairobi's street
food scene has created a critical knowledge gap, and addressing this gap will be critical in developing
targeted interventions.

The potential health effects of consuming contaminated street food require the identification of specific
microorganisms to quantify the associated risks. Therefore, the results will not only contribute to public
health policy and regulatory frameworks but also provide street vendors with knowledge to improve
their food safety practices. The project aims to improve overall food safety standards in Nairobi, protect
the health of city residents, and maintain the cultural importance of street food.

1.4 Objectives
1.4.1 Broad Objective
i. To assess and quantify the microbial quality of Nairobi Street food and identify potential health risks
associated with consumption.

1.4.2 Specific Objective


i. To isolate and characterize Fungi and Bacteria from street foods such as Roasted Maize, fried
Fish, sausage, and Banana.
ii. To determine the potential health risks associated with the consumption of street foods such as
Roasted Maize, fried Fish, sausage, and Banana.

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CHAPTER TWO: LITERATURE REVIEW
2.1 Overview of Street Foods
Street food is a dynamic part of a city's food landscape, reflecting the vibrancy and diversity of local
culture and cuisine (Hanser, 2021). These ready-to-eat or drinkable foods are typically prepared and sold
by vendors in public spaces, providing a convenient, affordable, and culturally significant alternative to
formal eateries. Street food markets in urban environments serve as lively gathering places where people
from all walks of life come together to enjoy a wide range of flavors and traditional foods (Allison,
2020). Street food includes a variety of dishes, from hearty snacks and simple meals to refreshing drinks
and desserts. This traditional recipes and innovative dishes, not only satisfies immediate hunger but also
provide a journey through the flavors and aromas unique to each location. Despite its popularity, the
safety and hygiene of street food have important considerations and require thorough research to ensure
consumer well-being in crowded urban environments.

2.1.1 Background of Street Foods


Street food has a rich and diverse background spanning centuries. This culinary tradition is part of many
cultures around the world, and it's a quick and easy way to enjoy a variety of affordable and delicious
dishes. Street food stalls can be found in crowded cities, lining the streets and attracting locals, tourists,
and foodies alike (Philosophers, 2023). The history of street food is closely connected to urbanization,
economic development, and exploration, making it an interesting topic to explore. The origins of street
food date back to ancient times, when small food stalls and stalls were set up outside markets and near
busy streets.

The migration of people from rural to urban areas increased the demand for quick and affordable meals,
and the ingenuity and creativity of street vendors responded to this need. Another important factor in the
development of street food is the influence of different cultures on culinary traditions. As explorers and
traders traveled across the continent, they encountered new ingredients, flavors, and cooking techniques
(de la Salle, 2019). These culinary exchanges have contributed to the formation of the variety of street
food we see today. Street food has a long and fascinating history that has evolved alongside urban
growth and cultural exchange. It plays an important role in providing affordable and diverse food
options to people on the go.

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2.1.2 Significance of Street Foods in Urban Settings
In urban settings, street food has been a staple for many years, and it holds such importance. The city's
busy streets have been transformed into a vibrant cooking hub, offering locals and tourists alike a variety
of affordable and delicious options. In addition to satisfying hunger, street food has several important
functions in urban areas (Moussavi et al., 2016). It contributes to the local economy, celebrates cultural
diversity, and promotes a sense of community and connection among city residents. The local economy
in urban areas is heavily influenced by the consumption of street food. Street food vendors often run
small businesses that not only earn a living but also create employment opportunities for others. These
microenterprises are part of the informal sector where entrepreneurs can start and operate businesses
with low start-up costs. Street food stalls stimulate economic circulation by purchasing ingredients from
local markets and employing local workers, contributing to the overall economic growth of the city.

Street food celebrates the cultural diversity that enriches urban environments. As cities become melting
pots of different cultures, street food serves as a gastronomic expression of this diversity. By consuming
these various goods, residents, and tourists can develop a greater understanding and respect for other
cultures in the city, which ultimately fosters tolerance and acceptance (Alfiero et al., 2019). Food stalls
serve as meeting places, facilitating social contact and fostering a sense of community among city
residents. The simple act of ordering from a street vendor often requires a short conversation that creates
an opportunity for interpersonal interaction. Whether it's sharing a table with a stranger or striking up a
conversation in line, food stalls are becoming a place for spontaneous encounters where people from all
walks of life come together (Nicula et al., 2018). This shared experience strengthens the social fabric of
urban communities and helps foster a sense of belonging among residents.

They support small businesses and contribute to the local economy by providing employment
opportunities. Street food celebrates cultural diversity and also serves as delicious ambassadors of
different culinary traditions. They promote community and connection among city residents; they also
serve as meeting places for people from all backgrounds to come together (Odundo et al., 2018). As
cities evolve and grow, vibrant street food scenes continue to be an integral part of the urban experience,
delighting the palate and inspiring a sense of togetherness.

2.1.3 Emergence of Concerns on Safety and Hygiene


Street food has become a source of concern due to the safety and hygiene concerns, which are now
commonly associated with this dish (Imathiu, 2018). Rapid urbanization, in combination with the

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informal nature of street food vending, has led to problems such as poor hygiene, inappropriate food
handling practices, and increased microbial contamination. The increasing availability of diverse and
convenient food choices on street corners is causing concerns about the safety of public health. Despite
the abundance of food options on the street, there are still many cases of people falling ill due to
foodborne illness that must be addressed through improved hygiene and safety measures (Sousa et al.,
2021). This recognition, in response to the evolving dynamics of urban food consumption, has led to
research initiatives and regulatory efforts aimed at improving food safety standards, promoting provider
education, and promoting safer street food environments.

2.2 Microbial Contamination in Street Foods


Microbial contamination of street food is widespread because street food sales are informal and
unregulated, posing a significant public health risk in urban areas. Pathogenic microorganisms such as
E. coli, salmonella, and norovirus can contaminate street food in a variety of ways (Bari & Yeasmin,
2018). This is caused, among other things, by contaminated water and unclean handling methods. This
contamination poses a variety of health risks, ranging from mild gastrointestinal upset to severe food
poisoning. Given the cultural importance of street food in urban environments, combating microbial
contamination is important to maintain public health.

2.2.1 Bacteria
Street food can harbor a variety of bacteria, depending on factors such as hygiene practices, food
handling, and environmental conditions. Escherichia coli (E. coli), a group of coliform bacteria, serves
as an indicator of fecal contamination and suggests poor hygiene during preparation (Were et al., 2020).
Staphylococcus aureus is known to produce toxins, can contaminate food when handled, and grows in
suboptimal storage conditions. Salmonella bacteria associated with food poisoning can be present in
contaminated meat and eggs used in street food. Listeria monocytogenes found in ready-to-eat foods
poses a risk of causing serious infections. Clostridium perfringens, which has been linked to food
poisoning, can grow in improperly stored prepared foods. Vibrio bacteria occur in seafood-based street
foods. If seafood is not cooked properly, it can cause infections. Campylobacter jejuni is common in
undercooked poultry and can cause gastrointestinal infections (Eromo et al., 2016). When combined
with unsanitary conditions, Shigella can cause gastrointestinal disease. Bacillus cereus, which can
produce toxins, is found in improperly stored starchy foods. Enterobacteriaceae, the bacterial family

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that includes coliform bacteria, can be indicative of poor hygiene or fecal contamination. To prevent
health hazards caused by street food bacteria, it is crucial to monitor the situation and implement strict
hygiene practices (Alimi, 2016)

2.2.2 Fungi and Molds


Rhizopus especially contributes to the rotting of fruits and vegetables. Alternaria spp., is associated with
fruits, vegetables, and grains and can produce mycotoxins. Mucor spp., associated with spoilage of
fruits, vegetables, and baked goods (Anwer et al., 2017). Fusarium spp. can contaminate grains, grains,
and legumes and can produce mycotoxins. Cladosporium spp. is a common outdoor contaminant found
in a variety of foods. The growth of these fungi and molds is influenced by factors such as temperature,
humidity, and the moisture content of the food. Although many molds are harmless, they can produce
mycotoxins, to prevent mold contamination in street food and maintain safety and quality standards in
urban food environments (Mailafia et al., 2017).

2.2.3 Viruses
Street food can contain a range of viruses and could be dangerous for consumers. Hepatitis A virus is of
particular concern because it can be transmitted through contaminated water and food, especially when
raw or minimally processed ingredients are involved. Norovirus is another serious viral threat known to
cause gastroenteritis and is often associated with ingesting contaminated food or water . The presence of
rotavirus, an enteric virus that is most commonly found in infants and young children, can pose a health
risk when street food contains potentially contaminated ingredients. Transmission of these viruses in
street food can occur through improper food handling, contaminated water sources, or contact with
infected food handlers. Protecting public health from viral infections requires implementing strict
hygiene controls, proper food preparation, and regular surveillance to reduce the risk of infection from
street food in various urban settings.

2.2.4 Parasites
The presence of multiple parasites can contaminate street food and pose health risks for consumers. E.g.
Protozoa such as Giardia lamblia and Cryptosporidium parvum are examples of parasites that can
contaminate street food, especially food that contains raw or undercooked food or contaminated water
sources (Barua et al., 2023). Helminths such as roundworms and tapeworms can also pose a threat if
they are found in undercooked meat or improperly processed foods Transmission of these parasites

7
through street food may be due to poor hygiene, improper cooking methods, or the use of contaminated
water in food preparation (Gemechu et al., 2022). Ensuring the safety of street food requires strict
adherence to hygiene standards, proper cooking procedures, preventing the transmission of parasitic
infections and protecting the health of consumers consuming these popular and diverse food sources in
urban environments.

2.3 Factors Contributing to Microbial Contamination


Microbial contamination of street food can be caused by poor hygiene practices among vendors, cross-
contamination in food handling, improper temperature control, use of contaminated water,
environmental factors such as dust and pests, and awareness and training among vendors (Birgen et al.,
2020). These factors collectively contribute to the risk of microbial spread in street food, highlighting
the need for comprehensive measures to improve food safety and reduce contamination risks in urban
environments.

2.3.1 Poor Personal Hygiene Practices


Poor personal hygiene practices among street food vendors are a significant risk factor for microbial
contamination of street food, with vendors not having access to proper handwashing facilities resulting
in poor hand hygiene (Were et al., 2020). Contamination in the hands can transfer pathogens to food
while it is being prepared or handled. Lack of protective clothing such as gloves and aprons further
increase the possibility of direct contact between the vendor's hands and the food. In crowded street
markets, where vendors are close to customers, respiratory hygiene measures are essential to prevent the
spread of microorganisms. Eliminating poor personal hygiene practices is essential to reduce the risk of
foodborne illness and improve the overall safety of street food in urban environments (Salamandane et
al., 2023). Educational efforts and regulatory measures play an important role in improving vendor
practices and minimizing potential health risks associated with poor personal hygiene when selling food
on the street.

2.3.2 Water Quality and Environmental Factors


The safety of food sold on street corners is largely dependent on the water quality and environmental
conditions. Contaminated water used for cooking and preparation of food is highly contaminated with
pathogens, which can easily transfer to the microbes that digest the food (Bereda et al., 2016). Poor air
quality, dust, and pests in outdoor stall environments affect overall environmental health. Improper
waste disposal and the presence of pests such as insects and rodents can further increase the risk of

8
contamination. To ensure food safety on the street, waste management is essential, as well as clean
drinking water for food preparation, and environmental control measures are necessary (Kariuki, 2018).

2.3.3 Equipment and Packaging Concerns


There are serious equipment and packaging problems involved when it comes to the safety and hygiene
of food sold on street corners. Utensils that have been poorly maintained can harbor microbial
contamination, which can occur when food is being cooked and exposed to microorganisms (Birgen et
al., 2020). Microbial contamination can be introduced by packaging that is not ideal or damaged, which
can compromise the food's integrity and increase the risk of contamination. When selling food on the
street, it is important to ensure proper cleaning and disinfection of equipment and the use of appropriate,
intact packaging materials to maintain food safety standards. The implementation of education and
training programs to educate vendors on the importance of hygiene practices and equipment
maintenance will help reduce these concerns and improve the safety and quality of street food in urban
areas.

2.3.4 Lack of Training and Awareness


Vendors in urban areas who are not trained and educated on food handling, hygiene practices for food
that kill bacteria, etc., or pollution control problems the vendor may have a greater impact than
otherwise. This knowledge gap can lead to suboptimal food safety standards and increase the risk of
microbial contamination of street foods (Desye et al., 2023). Training programs that focus on personal
hygiene, safe food handling, and proper hygiene are essential to provide street vendors with the skills
they need to reduce contamination risks. It is important to raise awareness about the importance of
adhering to food safety guidelines to promote a responsible culture and improve overall hygiene in street
food vending. Regulators and local governments are responsible for implementing educational programs
and providing resources to improve the training and awareness of street food vendors, ultimately leading
to safer and healthier street foods.

2.4 Microbial indicators of food quality and safety


Microbial indicators play an important role in assessing the quality and safety of street foods and serve
as important parameters for assessing contamination risks (Ma et al., 2019). The total microbial load and
the possibility of fecal contamination are determined by variables such as the total viable cell count
(TVC), coliforms, and Staphylococcus aureus, while the presence of certain pathogens like Salmonella
enterica and Listeria monocytogenes poses a significant health risk. Monitoring of mold, yeast, and

9
Vibrio bacteria is important for assessing both spoilage and contamination from seafood (Moloi et al.,
2021). Regular monitoring of these indicators is important to detect problems early, facilitate corrective
actions, and maintain safety standards in the diverse and dynamic context of street food vending.

2.4.1 Total Viable Count (TVC)


The use of Total Viable Count (TVC) is an essential method for measuring the total microbial load in
street food, which includes bacteria, yeast, and mold. This provides insight into overall food hygiene,
freshness, and safety and reflects the effectiveness of street vendors' food handling and storage methods.
Elevated TVC levels may indicate possible contamination or poor hygiene (Wiatrowski et al., 2023).
Monitoring TVCs in street food is important to ensure the microbiological quality and safety of food
served in urban environments. TVC evaluation enables early identification of risks and timely corrective
actions to maintain street food quality and safety standards for consumers.

2.4.2 Coliforms
Coliform bacteria, a group of Gram-negative rod-shaped microorganisms, serve as important microbial
indicators for assessing food safety and quality. Among them, Escherichia coli (E. coli) stands out as a
particular type used to measure fecal contamination and the possible presence of pathogens. Monitoring
coliform levels, including other species such as Enterobacter, Klebsiella, and Citrobacter, provides
insight into general hygiene in food processing and handling (Eden, 2014). Regular evaluation of
coliform bacteria serves as an early warning system that facilitates corrective action and ensures
compliance with hygiene standards in food production and handling. By serving as a reliable indicator,
coliform bacteria significantly contribute to maintaining food safety and quality and provides a proactive
approach to foodborne illness prevention and consumer welfare.

2.4.3 Standard Plate Count (SPC)


Standard blood cell count (SPC) is a commonly used microbial indicator to assess food safety and
quality. Determines the total number of viable aerobic microorganisms, such as bacteria, yeast, and
mold, present in a given sample (Eromo et al., 2016). SPC provides a general assessment of the
microbial load and general hygiene of foods. Elevated SPC levels may indicate poor hygiene, improper
handling, or improper storage conditions, which can pose a potential risk to food safety and quality.
SPC monitoring is important for several reasons. As a quantitative tool, SPC helps maintain optimal
hygiene practices throughout the food production and distribution chain, thereby contributing to the
delivery of safe, high-quality food to consumers.

10
2.4.4 Molds and Yeasts
Molds and yeasts are important microbial indicators for assessing food safety and quality. Their
presence in food can cause spoilage and, in some cases, the production of harmful mycotoxins. Mold
and yeast detection serves as a valuable tool for preventive measures and contributes to the overall
safety of food and the protection of consumer health.

2.5 Hygiene and Regulatory Challenges


The management of food safety in urban settings is complicated by both sanitary and regulatory
considerations in the context of street food vending. The informal and widespread nature of street food
sales often hinders the implementation and enforcement of strict hygiene standards (Nkosi & Tabit,
2021). Street vendors may encounter obstacles in meeting legal obligations due to limited resources,
expertise, or financial constraints. The situation is further complicated by the lack of adequate sanitation
and access to water for hygiene measures. The varied and dynamic nature of street food markets can
make it challenging for regulators responsible for ensuring food safety to adhere to regulations. When
cultural practices, economic considerations, and public health goals intersect, effectively addressing
sanitary and regulatory challenges requires a nuanced approach (Alfiero et al., 2019). Urban
environments require a collaborative effort between local governments, providers, and communities to
enhance hygiene practices and ensure food safety on the street, with specific education and regulatory
initiatives being implemented.

2.6 Global Perspectives on Street Food Safety


A global perspective on street food safety provides valuable insight into the various challenges and
practices surrounding this popular culinary phenomenon. Street food is an integral part of the food
culture of urban centers around the world, providing a convenient and culturally rich dining experience
(Kamala & Kumar, 2018). However, street food safety varies widely from region to region due to
differences in regulatory frameworks, hygiene standards, and socio-economic factors. While some
countries have strict regulatory measures in place to ensure the safety of street food, others find
enforcement and monitoring difficult, leading to potential health risks.

Despite the presence of regional differences, comparative studies on microbial contamination in street
food demonstrate that specific conditions require context-specific actions. Providing examples of
successful street food regulation and safety protocols in a specific area can be useful in guiding policy
initiatives to improve food safety in other areas (Desye et al., 2023). The integration of global initiatives

11
and endeavors contributes to a more extensive comprehension of street food safety and facilitates the
exchange of knowledge and best practices to ensure consumer well-being in diverse urban settings.

12
MATERIALS AND METHODS
Materials

1. Sterile containers for sampling


2. Cooler box for sample transport
3. Sterile gloves
4. Disinfectants for surface cleaning
5. Sterile tubes
6. Labels
7. Markers
8. Balance for accurate measurement (weighing)
9. Source of heat
10. Test tubes
11. Inoculating loops for transferring cultures and streaking plates
12. Nutrient Agar plates for culturing bacteria
13. Culture media – Nutrient agar for bacteria and Potato Dextrose Agar for fungal
14. Slides
15. Stains (Methylene blue or crystal violet)
16. Sabouraud dextrose agar (SDA) plates for fungal culturing
17. Petri dishes
18. Incubator for bacterial and fungal culture growth
19. Microscopes for microbial observation
20. Pipettes for accurate measuring
21. Autoclave for sterilizing equipment and media
22. Biosafety cabinet for aseptic work (the laminar flow)
23. Colony counter for microbial colonies
24. Refrigerator for storing samples and culture media
25. Biohazard waste container
26. Distilled water for preparing culture media and dilutions

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3.1 Study Area
The food samples studied were collected in the stalls within the City Market, CBD Nairobi town. The
stalls sold ready-to-eat food. The research was carried out in the laboratory of the University of Nairobi,
School of Biological Sciences.

https://maps.app.goo.gl/2gMp7Xs3i7ikyHCe6

Figure 1: Location of the study area

3.2 Sample Collection


Four food samples namely, banana, fried fish, sausage, and roasted maize (corns) were collected from a
random food stall that sold ready-to-eat food prepared on site. One of each warm sample was collected.
Collection was done between 1 PM and 2 PM: when the place was busy with consumers looking for their
meals. The samples were purchased in sellers’ packaging material then kept in sterile polythene bags and
transported to UON, Department of Biology laboratory in cooler boxes for microbial analysis. The
analysis was done within 2-3 hours of sample arrival and extra stored in a 4 °C refrigerator.

14
Figure 2: samples collected for the study

3.3 Media Preparation


3.3.1 Nutrient Agar
Nutrient agar served as the growth medium for assessing the antibacterial activity of plant extracts. Fifteen
grams of the dehydrated agar powder were aseptically weighed and added to one Liter of sterile distilled
water. The mixture was stirred continuously using a sterile stir bar to ensure complete solubilization of
the agar and achieve a homogenous suspension. To eliminate any potential microbial contaminants, the
prepared agar solution underwent autoclaving at 121°C for 15 minutes. Following autoclaving, the
solution was allowed to cool to approximately 45-50°C. This temperature range is optimal for handling
the agar without compromising the viability of subsequently introduced bacteria. Under aseptic
conditions, the cooled agar solution was carefully poured into sterile Petri dishes. This process ensured
minimal disruption to the sterility of the media. The agar was distributed evenly to create a uniform layer
at the bottom of each dish. After pouring, the plates were left undisturbed on a level surface to solidify.
To prevent excessive condensation that could dilute the media, the lids were placed slightly ajar during
this stage.

3.3.2 Potato Dextrose Agar


By using a weighing balance 39g of PDA powder was weighed and dissolved in 1 liter of distilled water.
The mixture was stirred to achieve complete dissolution and then autoclaved at 121°C for 15 minutes for
sterilization. The mixture is then let to cool to approximately 45-50°C. Aseptically, the sterilized agar was
15
poured into sterile Petri dishes, ensuring an even layer at the bottom. After pouring, the plates were left to
cool and dry with their lids slightly ajar to prevent condensation. Once dry, the plate's lids were closed
tightly and stored in a cool place to solidify. Throughout the process, the aseptic technique was maintained
to prevent contamination.

3.4 Isolation of Microorganisms


3.4.1 Serial Dilutions
For each sample, 1 gram was weighed and suspended in 9 milliliters of distilled water in sterile dilution
bottles. The suspensions were then mixed thoroughly to make stock solutions. By the use of a sterile
micropipette tip and a micropipette, 1 milliliter of the stock solution was transferred onto a dilution bottle
containing 9 milliliters of distilled water to form the 101 dilutions. The suspension was then mixed
thoroughly. The 1:10 serial dilutions were done up to 104 using the previous dilution as a stock solution
for the next dilution and sterile pipettes at each dilution.

3.4.2. Isolation of Bacteria on Nutrient Agar


Isolation was done inside a laminar flow. Two nutrient agar plates for each food sample were appropriately
labeled as 101 dilutions of the respective sample and 102 dilutions of the respective sample. On the nutrient
agar plates, 0.1 milliliters of 101 dilutions and 102 of each sample were pipetted. The samples were then
spread evenly on the surface of the agar using a glass rod sterilized by dipping in ethanol and flaming.
Sterile pipette tips were used for each dilution of each sample. The plates were incubated at 27°C for 24
hours.

3.4.3 Isolation of Fungi on the PDA


Following an aseptic technique near a flame and within a sterile chamber, Petri dishes labeled with food
samples of bananas, fried fish, sausage, and roasted maize were opened slightly. Sterilized forceps were
used to pick a small piece of banana and roasted maize, which were then placed at the center of the PDA
petri dish. For fried fish and sausage, a sterilized scalpel was used to scrape a small amount of material
and then placed onto the agar surface. Petri dishes were then gently closed and incubated at room
temperature for 72 hours.

16
3.4.5 Sub-culturing
A representative colony from each distinct colony group from the initial isolation of each sample was
picked for sub-culturing and was done on nutrient agar plates for the bacterial analysis and the fungal
analysis in PDA inside a biosafety cabinet. A sterilized wire loop is used to pick a piece of the colony
and transfer it to an appropriately labeled agar plate. The plate streak method was used and the inoculum
was spread evenly on the plate and incubated at 27℃ for 24 hours. For the fungi, it was incubated for 3
days.

3.5 Characterization of Isolates


3.5.1 Colony Morphology Characteristics
Isolates were characterized based on the characteristics of the colonies and gram staining. Colony
characteristics to be used include colony color, size, form, elevation, and margin.

3.5.2 Gram Staining


After Subculturing on nutrient agar plates to obtain well-isolated colonies. A representative of each colony
group was picked for gram staining. A small amount of bacterium was placed in a drop of water on a glass
slide and spread evenly using a sterile wire loop. The slide was air-dried to form a smear and then passed
over a flame (heat-fixed). The prepared smear was then stained with the primary dye; crystal violet for 60
seconds. Then the stain was poured. The smear was covered with Gram’s iodine for 60 seconds and then
rinsed with distilled water. The stained smear was then rinsed with 95% ethanol one drop at a time, holding
the slide at 45℃angle. Ethanol was then run until there was no blue or purple color release and then rinsed
immediately with water. The smear was then covered with safranin for 30 seconds and was then rinsed
with distilled water. It was then blot-dried with a tissue paper. Care was taken to avoid decolorizing the
smear for too long.

17
CHAPTER FOUR: RESULTS

4.1 Bacterial Analysis


4.1.1 Isolated Bacteria on Nutrient Agar

Figure 3: Isolated bacteria from fish Figure 4: Isolated bacteria from sausage

Figure 5: Isolated bacteria from Roasted maize Figure 6: Isolated bacteria from banana

18
4.1.2 Subculture bacteria

Figure 7: Subculture of bacteria isolated Figure 8: Subculture of bacteria isolated


from fried fish from Sausage

Figure 9: Subculture of bacteria isolated Figure 10: Subculture of bacteria isolated from
Roasted Maize Banana

4.1.3 Morphological Characterization of the Isolates

19
Isolates color Size Shape Margin Elevation Texture Gram
Reaction
1 White Large Irregular Wavy Umbonate Shiny Positive

2 White Small Circular Entire Convex Shiny Negative

3 White Small Circular Entire Convex Shiny Negative

4 Colorless Puncture Irregular Wavy Umbonate Rough Positive

Table 1: Morphological characteristics of Bacterial isolates.

4.1.4 Gram Stain

Figure 11: Bacillus sp. Figure 12: Streptococcus sp.

Figure 13: E. coli

4.2 Fungal Analysis


4.2.1 Isolated Fungi on PDA

20
Figure 14: Fungi isolated from Roasted Maize

Figure 15: Fungi isolated from Sausage

Figure 16: Fungi isolated from Banana

21
Figure 17: Fungi isolated from Fish
4.2.2 Characterization of Isolated Fungi

Figure 18: Sub-culture of Fungi Isolated from Fried Fish

Figure 19: Sub-culture of Fungi Isolated from Sausage


22
Figure 20: Sub-culture of Fungi Isolated from Roasted Maize

Figure 21: Sub-culture of Fungi Isolated from Banana

23
4.2.3 Microscopic Characterization of Isolated Fungi

Figure 22: Fusarium sp. Isolated from Fish Figure 23: Trichophyton tonsurans sp. Isolated
from sausage

Figure 24: Fusarium sp. Isolated from Roasted Figure 25: Fusarium sp. Isolated from Banana
maize

24
4.2.4 Morphological Characterization of the Isolated Fungi

Food Sample Growth Rate Diameter of Pigmentation Probable Fungi


Colony in PDA during Growth
Roasted Maize Fastidious 6cm White to black Fusarium

Banana Fastidious 6cm Yellow to White Fusarium

Fish Moderate 3 cm White Aspergillus

Sausage Moderate 3 cm White Trichophyton


tonsurans sp.

Table 2: Table 3: Morphological characteristics of Fungal isolates.

25
CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATIONS
5.1 Discussion
The laboratory tests revealed that all the food samples from the are contaminated with different bacteria
and fungi species. These include Staphylococcus aureus, Streptococcus sp, Bacillus sp, Flavobacterium
sp, Escherichia coli, Aspergillus flavus, Aspergillus niger, Penicillium sp, Fusarium sp. The finding is in
agreement with the work of Ajao and Atere (2009) and Oranusi and Braide (2012). The presence of
Staphylococcus aureus in the samples is indicative of human contamination after production. This could
be from direct human contact such as through fingers or indirectly through utensils. The organism is
associated with endotoxin characterized by a short incubation period (1-8 hours), violet nausea, vomiting,
and diarrhea. Similar to findings reported by Njage et al. (2019), the study identified high levels of total
coliforms and Escherichia coli in street foods, indicating fecal contamination and poor hygiene practices
during food handling (Njage et al., 2019). Contrary to the study by Osaili et al. (2018), which found lower
prevalence of Streptococcus spp in street foods, the study observed higher counts of this pathogen,
suggesting variability in contamination levels accross the City Market (Osaili et al., 2018).

Consistent with the findings of Hoque et al. (2020), Bacillus spp. Were detected in a considerable
proportion of street food samples, highlighting the potential risk of foodborne illness associated with
consumption of contaminated foods (Hoque et al., 2020). Bacillus cereus is another isolate that is
associated with the production of toxins, diarrheal, and emetics in food which causes food poisoning. It is
found in dust, soil, and raw food and can survive normal cooking temperatures as a heat-resistant spore
(Rajkowski and Bennett, 2003). The presence of E. coli suggested fecal contamination. Although E. coli
is harmless, Enterohaemorrhagic E. coli (EHEC) is capable of producing one or more toxins and a
particular serotype O157:H7 have been associated with hemolytic ureamic syndrome and thrombosis.

The Enterotoxigenic E. coli (ETEC) is associated with traveler’s diarrhea similarly shigella dysenteriae
has been associated with severe bacillary dysentery while streptococcus sp, has frequently been associated
with acute sore throat (Adams and Moss, 2008). The presence of Aspergillus flavus, Aspergillus niger,
penicillium sp and Fusarium sp in the food sample is not surprising as they disperse in the form of spores
which is abundant in the environment and can be introduced through dust and soil (Apinis, 2003). Their
presence in these food samples is a serious public health concern as these fungi have all been implicated
in the production of Mycotoxin (Makun et al. 2009). as the study identified Aspergillus spp., Trichophyton

26
spp and Fusarium spp. As predominant fungal species in street foods, consistent with findings reported by
Rai et al. (2017) in similar settings (Rai et al., 2017).

In contrast to the study by Udomkun et al. (2019), which reported lower prevalence of fungal
contamination in street foods, the findings suggest a higher risk of fungal growth, possibly due to
differences in sampling methodologies and environmental conditions (Udomkun et al., 2019). Aspergillus
species produce toxic byproducts (mycotoxins) these toxins cause illness when consumed in high
quantities. Improper storage conditions, particularly warm and humid environments, promote Aspergillus
growth on food. Some fusarium species produce mycotoxins and spoilage of grains and fruits. The
presence of these bacteria and fungi indicates potential hygiene issues throughout the street food
preparation process. The major areas of concern are: Contaminated water source: use of unclean water for
washing ingredients or utensils can introduce E. coli and other harmful microbes. Poor hygiene: vendors
not properly washing their hands can spread bacteria from their bodies to food Improper temperature
control: if cooked food is not kept hot enough or low ingredients are not stored cold enough, bacteria can
multiply rapidly. Cross-contamination: using the same utensils for raw and cooked food can transfer
bacteria from contaminated items to safe ones.

Microbial guidelines for cooked food stipulated that the plate count must be <107 cfu/g for fish <1.0 x 104
cfu/g for banana <1.0 x 105 cfu/g for sausage and roasted maize <1.0 x 104 cfu/g and for coliforms the
plate count must be <10cfu/g (Gilbert et al. 200).

The microbial load of the food samples was higher than stipulated hence their presence constituted a health
risk. It can be adjudged that the street food sold in most streets in Nairobi City Market as obtained are
neither bacteriological nor mycological fit for consumption. The food contamination may have been due
to unhygienic practices and prolonged exposure to the environment.

5.2 Conclusion.

The findings of this study show the significant microbiological and mycological risks associated with
consuming street foods from City Market, Nairobi. The presence of pathogenic bacteria such as
Escherichia coli, Staphylococcus aureus, and Salmonella spp., along with potentially toxigenic fungal
species, highlights the urgent need for interventions to improve the safety and quality of street foods.

27
Several factors contribute to the contamination of street foods, including inadequate hygiene practices
during food preparation, improper storage conditions, and environmental contamination. The high
prevalence of microbial and fungal contamination across all food items examined indicates a systemic
issue that must be addressed to safeguard public health. To mitigate the risks associated with street foods,
multifaceted approaches are necessary. Strengthening food safety regulations and enforcement
mechanisms is paramount to ensure compliance with hygiene standards among street vendors. Regular
monitoring and surveillance of street food establishments should be conducted to detect and address
potential sources of contamination promptly.

Comprehensive food safety education programs targeting both street vendors and consumers are essential.
Vendors need to be trained in proper food handling practices, including hand hygiene, safe food storage,
and equipment sanitation. Consumers should be informed about the risks associated with consuming
contaminated street foods and empowered to make informed choices about their food purchases. Good
partnership with the government authorities, public health agencies, academia, and the food industry is
crucial to implement effective interventions and sustainably improve the safety of street foods in Nairobi.
Additionally, investments in infrastructure, such as providing access to clean water and sanitation facilities
for street vendors, can contribute to reducing the risk of contamination.

Addressing the microbiological and mycological hazards associated with street foods requires a concerted
effort from all stakeholders. By prioritizing food safety and implementing evidence-based interventions,
we can protect public health, promote consumer confidence, and ensure the availability of safe and
nutritious street foods for urban populations in Nairobi and beyond. Most food pathogens are of soil or
intestinal origin and are transmitted through poor food preparation, personal hygiene, or public sanitation
practices. Therefore, to ensure the safety of these foods, producers and hawkers must maintain a high level
of personal hygiene, and a clean environment, minimize contact with food samples after production, and
also utensils should be properly cleaned at all stages of production.

5.3 Recommendations
• Improved hygiene: street food vendors need training on proper hygiene practices including
handwashing, utensil sanitation, and proper food storage to minimize contamination
• Temperature control: maintaining proper cooking temperatures can kill harmful bacteria keeping
cooked food hot and perishable ingredients cold can help prevent bacterial growth.

28
• Source tracking: investigating the source of contamination can help identify the critical control points
in the food preparation process.
• Public awareness: educating consumers about the potential risks of street food and how to identify
vendors can empower informed choices.

29
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