Republic of the Philippines

Department of Education
Caraga Administrative Region
Bayugan City Division
BAYUGAN NATIONAL COMPREHENSIVE HIGH SCHOOL

Microbiological Assessment of street foods in the Nightmarket of

Bayugan City

In Partial Fulfillment of The Requirements

In Research Subject 10

CHRIS CLARENCE R. TAGUNO *

REAN ZOE G. TERIOTE
GIEMUEL HARRIS P. TAMPON

Researchers

Vincent Butch S. Embolode

Research Adviser

(2023).-2024
 CHAPTER I
INTRODUCTION

1.1 Background of the study

2.5 billion people worldwide are estimated to eat Street Foods every day

(FAO & WHO, 2022). Street food stands are common throughout most of Asia

(Jayasuriya, 1994). Street food is becoming increasingly popular, with new

cuisines emerging and gaining popularity, led by the Caribbean, Venezuelan,

and Japanese styles of cooking (General, 2022). While street foods are

appreciated for their unique flavors and convenience, these are also often

perceived to be a major public health risk due to unsafe food handling (FAO &

WHO, 2022). A significant public health risk factor is the ignorance of street

food vendors regarding the origins of foodborne illnesses. (FAO & WHO, 2022).

Educational status, training, vending experience, and monthly income have

all been identified as determinants of street food vendors with food safety

compliance (Negassa et al., 2023) . Regulations on street food vending vary

from country to country and even city to city (Wheels, 2021).

Street Foods in the Philippines play vital role in providing and ensuring

the residents a variety of different foods that are easily accessible and are

quickly prepared for the consumers. One thing that is unrealized by most of

the residents are the health and safety of the foods that the residents are

buying and consuming, unawareness of the food safety may cause diseases,

illnesses to the consumers due to the food that these consumers are eating.

The estimated amount of diseases that are caused by eating food tainted with

bacteria, virus, parasites or chemical substances such as heavy metals are

more over 200 diseases ranging from diarrhea to cancers (World Health

Organization: WHO, 2022).

In some local cities, such as Bayugan City, may consist of an urban

area with street food stands with street food vendors, just like the Bayugan

City Night Market. The Bayugan City Night Market attracts many consumers

due to it being positioned at a public and easily accessible area, and also there

are many different variety of Foods that are sold, such as Kwek-Kwek, Fish

Balls, Tempura, Hotdogs, Siomai on sticks, BBQ, etc. Based on the

Researcher’s observations, foods such as Kwek-Kwek, fish balls, tempura,

hotdogs and siomai on sticks are normally displayed out for the public,

leaving it exposed to potential bacteria and viruses. Salmonella spp.,

Clostridium perfringens, Escherichia coli, Shigella spp., Campylobacter jejuni,

Staphylococcus aureus, Bacillus cereus, and other foodborne pathogens are

frequently linked to street-vended foods (Birgen et al., 2020), however the

researchers aim to identify the abundance of the three following bacteria

namely Salmonella, Staphylococcus aureus, and E. coli (Escherichia coli)

present in different samples of street foods from the Bayugan City Night

Market.

E. coli (Escherichia coli) is a type of bacteria that often resides in your

intestines. Some animals' guts contain it as well. Nearly all E. coli are safe and

even aid in the maintenance of a healthy digestive system. However, some

strains might result in diarrhea if a person consumes infected food or drinks

tainted water. Despite the fact that many individuals associate E. coli related

to food poisoning, other bacteria-related illnesses include pneumonia and

urinary tract infections. In actuality, E. coli is the root cause of 75–95% of

urinary tract infections. E. coli is a typical colon inhabitant, and this is how it

enters the urinary tract. Some versions of E. coli make an individual sick by

making a toxin called Shiga. This toxin damages the lining of an individual’s

intestine. (Aaron Gould Sheinin et. al 2017)

The water used by street vendors may contain bacteria and

microorganisms such as E. coli, as it is not always filtered properly. E. coli

might have been present in the contaminated water used by food handlers, as

street vendors reuse water repeatedly even when it is no longer clean to use

(Moloi et al., 2021).

The causative E will be used to characterize intestinal diseases. Shiga

toxin-producing Escherichia coli (STEC), also known as enterotoxigenic

Escherichia coli (ETEC), enterohemorrhagic Escherichia coli (EHEC), which

will be referred to as EHEC/STEC, and enteroinvasive Escherichia coli (EIEC),

enteropathogenic Escherichia coli (EPEC), and enteroaggregative Escherichia

coli (EAEC). (Mueller, 2023).

The most harmful of the numerous common staphylococcal bacteria is

Staphylococcus aureus. These gram-positive, sphere-shaped (coccal) bacteria

can also cause pneumonia, heart valve infections, and bone infections. Some

medications may not be able to treat these illnesses. (Bush, 2023)

S. aureus is most often spread to others by contaminated hands.

Normally, the skin and mucous membranes act as a strong barrier to prevent

infection. But in the event that these defenses are compromised (for example,

by trauma-related skin damage or viral infection-related mucosal damage), S.

Infection can result from aureus getting into the bloodstream or underlying

tissues (About Staphylococcus Aureus - MN Dept. Of Health, n.d. 2010). The

presence of Staphylococcus aureus in food is usually due to cross-

contamination of ready-to-eat food with either raw food or contamination from

a food handler that is carrying Staphylococcus aureus. The bacteria can

multiply in the food and produce toxins that can make people ill (CDC 24/7,

2023). Staphylococcus aureus bacteria cause skin infection. This may cause

your skin to get red, blister, and boil. These infections can be anywhere on an

individual’s body, including the face, often around the mouth and nose. Even

if you are healthy, Staphylococcus bacteria commonly live in your nose or on

the skin. If the bacteria get inside your body, these can cause problems

(Professional, 2023.). When the bacteria do, these can create many thousands

of serious cases of S. aureus infections in the U.S. every year. People who are

hospital employees or patients have higher percentages. Direct touch, contact

with contaminated things (including gym equipment, phones, doorknobs,

television remote controls, or elevator buttons), or, less frequently, contact

with infected droplets dispersed by coughing or sneezing are all ways that the

bacteria might transfer from one person to another. People that carry the

germs but do not exhibit any symptoms from it are considered carriers.

Carriers can spread bacteria from their nose to other regions of their body

with their hands, which can occasionally result in infection. (Bush, 2023)

Food handlers who don't wash their hands, as well as the tools and

surfaces the food handlers use in between food preparation procedures and

when consumers consume raw or undercooked food, can potentially spread

Salmonella. Additionally, animals and humans can contract Salmonella

(Nutrition, 2019). The transmission of Salmonella to street foods is a complex

process influenced by various factors, including the behavior and practices of

food vendors, the microbiological quality of raw ingredients, and the

environmental conditions in which the foods are prepared and sold. Poor

personal hygiene, improper handling and storage practices of foods, and

inadequate knowledge of food vendors towards foodborne diseases are

associated with the transmission of Salmonella to street foods (Mohammed et

al., 2020)

Salmonellosis is a bacterial infection caused by Salmonella. It ranges

clinically from the common Salmonella gastroenteritis to enteric fevers,

including typhoid fever. When there is no apparent explanation for an acute

diarrheal or feverish disease, it is important to rule for salmonellosis. By

separating the organisms from clinical specimens (blood or stool), the

diagnosis is verified. There are effective typhoid fever vaccines available, but

not for salmonellosis that is not typhoidal. Those diseases are controlled by

hygienic slaughtering practices and thorough cooking and refrigeration of food

(Giannella, 1996).

Eating raw or undercooked meat, poultry, eggs, or egg products, as well

as drinking unpasteurized milk, are the usual causes of Salmonella infection.

The interval between exposure and disease, or incubation period, can range

from six hours to six days. Often, people who have salmonella infection think

that these people have the stomach flu (Mayo Clinic, 2022).

A Salmonella infection often lasts a few days to a week in terms of signs

and symptoms. Although diarrhea may continue up to 10 days, it may take

many months for bowel movements to return to normal. Typhoid fever, a

frequently fatal illness that is more prevalent in developing nations, is caused

by a small number of salmonella bacteria species. (Mayo Clinic, 2022).

Salmonella infections can cause diarrhea, cramping in the stomach

(abdominal pain), fever, nausea, vomiting, chills, headaches, and blood in the

stool (Mayo Clinic, 2022).

 Pathogens such as Salmonella, Staphylococcus aureus, and E. coli are

present in street foods due to various factors related to food handling and

preparation. Contamination can occur during food preparation, post-cooking,

and various handling stages. For example, improper food handling practices

by food vendors, such as reusing surfaces without cleaning, not wearing

aprons, and carrying pathogens like S. aureus, can contribute to the presence

of these microorganisms in street foods (Moloi et al., 2021). Additionally, the

presence of pathogenic organisms such as E. coli is a cause for concern as it

usually indicates recent fecal contamination, posing a serious health problem

to the community (Kariuki, 2018). Furthermore, the lack of knowledge of food

hygiene among street vendors and the absence of regular inspection and

monitoring contribute to the presence of these pathogens in street foods

(Elobeid, 2014).

This research aims to determine the abundance of the three following

bacteria namely Salmonella, Staphylococcus aureus, and E. coli (Escherichia

coli) present in different samples of street foods from the Bayugan City Night

Market.
1.2 Statement of the problem

This study aims to determine the microorganisms present in street foods sold

in Bayugan City. Specifically, this study seeks to:

1. determine the bacterial species present in street foods; Salmonella,

Staphylococcus aureus, and E. coli (Escherichia coli)

2. determine the phenotypic characterization of microorganisms present in

street foods, through E. coli Count, Salmonella sp. Detection, Staphylococcus

aureus Count.

3. determine the quantitative and qualitative characteristics of the presence of

microorganisms in street foods such as:

a. most probable count (MPN) of coliforms

b. immunoassay for detection of enterotoxins and toxins of pathogenic

bacteria
 1.3 Conceptual Framework

Input Output
Independent Variable: Dependent Variable:
Process
Street Food samples Abundance of the three
common bacteria
Salmonella,
Staphylococcus aureus,
and E. coli

Microbiological testing on
street foods

Parameters

 E. Coli count
 Salmonella sp.
Detection,
Presumptive
(conventional)
 Staphylococcus
aureus count

The figure shows the variables that will be included in the study. The
dependent variables will be the selected street foods that will be tested. The
independent variables are the abundance of the three common bacteria
(Salmonella, Staphylococcus aureus, and E. coli). For the process, the street
foods will be run on a microbiological test in order to determine the possibility
of these bacteria contaminating the food. The parameters are E. Coli count,
Salmonella sp., and Detection, Presumptive (conventional).
1.4 Significance of the Study

The following groups will benefit from the study’s findings.

Consumers. The findings of this study will redound to benefit the consumers

by being aware of the potential health risks associated with consuming street

foods and taking necessary precautions to avoid foodborne illnesses since the

study can show the consumers the overall microbiological safety and quality

of the street foods from the Bayugan City Night Market.

Street food vendors. The vendors within the Public Night market would be

informed of the microbiological safety and quality of their street foods so that

the vendors will know about the proper sanitation of utensils and tools used

in preparing street foods, protecting their food from dust, and proper handling

of food.

Community Health Office. The findings of this study will help the city health

office assess the situation of the Public night market, which would allow the

community health office to develop guidelines and regulations for street food

vendors to follow to ensure the safety of the food that the vendors sell in the

food stalls of the Bayugan City Public Night Market.

Future Researchers. The findings of this study will benefit future researchers

as future researchers can use the information from this study to propose and

implement measures to improve food safety and hygiene in the Bayugan City

Night Market. Future researchers can also use the data from the findings of

this studyto create awareness among consumers about safe food handling

practices and potential risks associated with consuming street food.

1.5 Scope and Limitations of the Study

The study is to be conducted in the night market of Bayugan City,

Agusan del Sur, evaluating the food microbiological safety and quality issues

of street foods. This study will focus on street food’s microbiological safety and

quality by determining the abundance of the three kinds of bacteria found in

street foods namely Salmonella, Staphylococcus aureus, and E. coli

(Escherichia coli). The study will only include the street food vendors within

the vicinity of the Bayugan Night Market, only the regular street food vendors

in the night market will be included.

Map showing the location and outline of the study area.

The photo is from google maps
https://www.google.com/search?
rlz=1C1CHBD_enPH922PH922&tbs=lf:1,lf_ui:4&tbm=lcl&sxsrf=AJOqlzXwAurZB7MqIhnJn54DGvdTIxuuIw:1
673354718628&q=Bayugan+City+Night+market+google+maps&rflfq=1&num=10&ved=2ahUKEwjO7diMhL38
AhUOQd4KHR7bCooQtgN6BAgQEAY#rlfi=hd:;si:;mv:[[8.718778877713484,125.75624092965948],
[8.713794561967786,125.74609145074713],null,[8.716286728150147,125.7511661902033],17]

1.6 Key Terms and Definitions:

• Abundance: The quantity or presence of microorganisms.

• Carriers: Individuals who carry Staphylococcus aureus without displaying

symptoms and can potentially spread the bacteria to others.

• Coagulase Test: A test to detect the enzyme coagulase produced by some

Staphylococcus species.
• Community Health Office: A government office responsible for public
health in a community.

• Consumers: Individuals who purchase and consume street foods.

• E. coli (Escherichia coli): A type of bacteria commonly found in the

intestines of humans and some animals, with most strains being harmless.
However, some strains can cause food poisoning and other illnesses.

• Food Safety Compliance: Adherence to regulations and practices that

ensure the safety of food products.

• Foodborne Illnesses: Diseases caused by consuming food contaminated

with harmful bacteria, viruses, parasites, or chemicals.

• Halo-Tolerant Bacteria: Bacteria that can tolerate high salt concentrations.

• Hemolytic Test: A test to determine if Staphylococcus species can break

down red blood cells.

• Immunoassay: A test that uses antibodies to detect specific molecules, often

used to detect toxins of pathogenic bacteria.

• Microbiological Safety and Quality: Refers to the safety and characteristics

of microorganisms in street foods.

• Microorganisms: Tiny organisms such as bacteria and viruses.

• Most Probable Count (MPN) of Coliforms: A statistical method for

estimating the concentration of coliform bacteria in a sample.

• Phenotypic Characterization: The study of observable characteristics of

microorganisms.

• Public Health Risk: A potential threat to the health of a community or

population.

• Regulations: Rules and guidelines set by authorities to govern the sale and
preparation of street food.

• Salmonella: A genus of bacteria commonly found in the intestines of

humans and animals, often causing gastrointestinal infections.

• Salmonellosis: A bacterial infection caused by Salmonella, leading to

symptoms such as abdominal cramps, fever, and diarrhea.

• Shiga Toxin: A toxin produced by certain E. coli strains, such as O157:H7,

which can lead to severe symptoms including bloody diarrhea, vomiting, and
abdominal cramps.
• STEC (Shiga Toxin-Producing E. coli): A group of E. coli strains that
produce Shiga toxin and can cause severe illnesses.

• Staphylococcus aureus: A type of bacteria responsible for various

infections, including skin infections, pneumonia, and bone infections.

• Street Food Vendors: Individuals who sell food on the streets or in public
places.

• Street Foods: Ready-to-eat foods sold on the streets or in public places,

often characterized by their convenience and unique flavors.

• Transmission: The process of spreading bacteria through direct contact,

contaminated objects, or inhaling infected droplets.

• Typhoid Fever: A severe illness caused by specific Salmonella bacteria

species, more prevalent in developing nations.

• Urinary Tract Infections: Infections that affect the urinary system, often
caused by E. coli bacteria.
 Chapter II
Review of related literature and review of related studies

2.1 Introduction
This chapter contains the relevant literature, Research, thoughts, and
statements that give information about the study and a thorough description
of the ideas.

2.2 Related Literature

Preparation, Contamination and Health Risks of Street Food

A study was conducted in Vitória and Serra, Espírito Santo, Brazil,

about analyzing the handling and selling circumstances of street food.

Samples of the food sold in these trades were obtained legally in order to

assess the microbiological purity of the food. After 200 deals were reviewed, it

was found that "Poor" cleanliness was present in 58.5% of the sellers. The

main flaws were the seller's sanitary standards. After 63 food samples were

examined, it was determined that 57.1% of the vendors had contamination

levels higher than permitted, which made the food samples dangerous to eat.

(Ferrari, et al. 2021).

Meva'a et al. (2000) report that in Daloa, a Côte d'Ivoire urban area,

street food poses a growingly concerning health risk. It is also frequently seen

that the methods such as recylcling used frying oils and indgredients (tuna,

attiéké, fresh veggies, and oil) used in the preparation and serving of "garba,"

a popular street-food dish, exceed the Ivory Coast's food hygiene standards

and can be a possible source for health risks for consumers, (Bhattacharya et

al., 2008; Soula et al., 2020; Koffi, A. L. J. 2021).

Studies with similar samples

In the study about Street Food’s Microbiological Load and Vendors’ Food

Hygiene and Safety Practices Compliance in the Schools of Cebu City a cross-

sectional analytical study and descriptive-correlational research design was

employed. The food samples (kwekwek and two kinds of dipping sauce) for the

cross-section analytical study were purchased from street vendors in Cebu

City's north and south, and the food samples were examined in a lab for signs

of Escherichia coli bacteriological contamination. The food hygiene and safety

practices compliance of street food sellers were examined for the descriptive-

correlational research by looking into the characteristics of a certain

community. The E. coli laboratory tests' count revealed that every food sample

from every location indicated fecal contamination. According to the results of

the compliance scores, food vendors from the chosen study areas have very

good compliance with using protective clothing, having water available at the

vending point, cleaning their fingernails, and using the proper kitchenware

when serving food to customers. However, the food vendors have very poor

compliance with using a head covering, getting a business permit, and getting

a sanitary/health card. Despite the study's somewhat positive total

compliance score, the findings may indicate that there is no connection

between the vendors' standards for food safety and cleanliness and the

presence of E. coli in the examined food samples (Origenes, 2022).

In another study about Microbiological contamination and evaluation of

sanitary conditions of hot dog street vendors in Southern Brazil the

microbiological quality and sanitary conditions of hot dog vendors of Southern

Brazil were evaluated. Twenty hot dog samples were examined for coagulase-
positive staphylococci and total and fecal coliforms. A questionnaire consisting

of five blocks with questions about food handlers, installations and

equipment, raw materials and ingredients, food preparation and storage, and

documentation was used to analyze the hygienic conditions of vendors. The

findings showed that 75% of the hot dogs had total coliform contamination,

30% had fecal coliform contamination, and 25% had coagulase-positive

staphylococci levels higher than those allowed by Brazilian law. The majority

of merchants defrosted sausages at room temperature or with insufficient

cooling, according to survey results. Not a single vendor had a thermometer,

and several vendors were using non-potable water. Lack of time and

temperature controls, the use of substances with unknown origins, and the

absence of cross-contamination prevention measures were other common

deviations (Kothe et al., 2016).

In another study about Microbiological status of various foods

served in elementary school based on social economic status differences in

Karawaci Region, Tangerang District – Indonesia A microbiological test was

carried out on fifteen different types of food. Rice with chicken, meet ball,

poffertjes, and noodle with chicken were among the samples taken from

Elementary School A. The fried breaded vegetables, banana chocolate, french

fries, burgers, and chicken noodles with noodles were among the samples

taken from Elementary School B. The final items were tempura, fried banana,

batagor, dumplings, and burgers from primary school C. Staphylococcus

aureus (SA), yeast and mold (YM), total aerobic bacteria (APC), and coliform

bacteria were subjected to quantitative analysis, whilst a qualitative test was

carried out for Salmonella sp. Dumplings were found to have the highest level

of contamination, with an APC level of about 105 cfu/g. Salmonella spp. are

present. was only discovered in meals acquired from C Elementary School.

Additionally, an Atomic Absorption Spectroscopy (AAS) metal contamination

test was conducted as part of this investigation, and the results showed that

no dangerous metal was detected to have contaminated any of the different

foods (Adolf et al., 2012).

Microbiological studies about street foods

Street food is characterized as prepared and/or sold by vendors and

hawkers, typically in public areas like streets and other comparable public

spaces. However, microbial food borne infections constitute a significant

public health issue connected to street foods. Since the antibiotic resistance

determinants can be transferred to other pathogenic bacteria and could

potentially affect the treatment of severe bacterial infections, food

contamination with antibiotic-resistant bacteria can therefore pose an

important risk to public health (Eromo et al., 2016). More than 200 types of

diseases are estimated to be caused or spread by food, occasionally causing

long-term health problems in vulnerable groups such as the elderly, pregnant

women, children and immunocompromised people (WHO, 2019).

In the study about Bacteriological Quality of Street Foods and Antimicrobial

Resistance of Isolates in Hawassa, Ethiopia A cross-sectional study was

conducted from May to September 2014. A total of seventy-two samples were

gathered from six different food items: cooked potatoes, raw fish, avocado,

chili, and local bread. Using normal microbiological procedures, bacterial

isolation, colony counts, and antibiotic susceptibility testing were performed.

The overall colony counts in almost 31% of the food samples ranged from

1.7x105 to 6.7x106 colony-forming units per gram (CFU/g), exceeding the

permitted limits for the microbiological quality of foods that are ready to be
eaten. In raw fish, "kita" and "ambasha," the mean coliform and

Enterobacteriaceae numbers were similarly higher than the maximum limits.

The most common isolates were E. Coli, Salmonella species, and S. aureus.

Confirming a high rate of contamination in Hawassa City's street-vended food.

The isolates of antibiotic-resistant bacteria and foodborne bacteria that have

been found may be dangerous to the community's health. Thus, it is advised

that vendors receive regular inspections, health education, and training on

safe food handling procedures. (Eromo et al., 2016).

In the study about the Contamination of ready-to-eat street food in

Pakistan with Salmonella spp 320 food samples, including sandwiches,

shawarma, pizza, and burgers, were purchased from street vendors. A total

aerobic count was acquired by plating samples on bacterial growth medium,

and this count was used to measure the amount of microbial contamination of

individual food samples between January and December 2018. Types of

Salmonella. Polymerase chain reaction was used to identify serovars. RTE food

samples had about 38% of items that were dangerous for human

consumption. Shawarma was the RTE food type that was most contaminated.

Summer showed a higher level of microbial contamination in food samples

than the other seasons. About 40% of food samples that were unfit for human

consumption were contaminated by Salmonella spp. The only serovars found

in the samples that tested positive for Salmonella spp. were S. enteritidis and

S. typhimurium. Of the 49 samples with high microbial counts, 34 samples

had S. enteritidis and 15 samples included S. typhimurium. The results of the

antibiotic sensitivity showed that both S. enteritidis as well as S. typhimurium

showed amoxicillin resistance. Furthermore, S. S. enteritidis was resistant to

erythromycin and chloramphenicol. typhimurium showed a significant level of

erythromycin resistance. Each S. S. typhimurium and S. Enteritidis had high

resistance with kanamycin (Raza et al., 2021).

In the study about Microbial levels on street foods and food preparation

surfaces in Mangaung Metropolitan Municipality samples were collected

through convenience sampling. Surface samples from certain media were

taken using swabs, and the samples were quantified. Along with the collection

of eight samples of beef, a RapID kit was used to quantify and identify the

microbial burden on each sample. In comparison to surface swabs collected in

Bloemfontein and Thaba Nchu, those from Botshabelo displayed greater

microbial counts. By comparison with international microbiological standards,

meat samples from Thaba Nchu, Bloemfontein, and Botshabelo showed higher

bacteria counts. The microbiological levels were evaluated, and the following

species were found: Peptostreptococcus tetradius, Corynebacterium jeikeium,

Candida guilliermondii, Escherichia coli, and Staphylococcus aureus (Moloi et

al., 2021).

In the study about bacteriological profile, antimicrobial susceptibility

patterns of the isolates among street vended foods and hygienic practice of

vendors in Gondar town, Northwest Ethiopia a cross-sectional study was

conducted. A standardized questionnaire was utilized to gather data on the

hygiene practices and socio-demographic traits of 24 vendors. Using the

standard aerobic plate count method, a total of 72 food samples representing

four distinct food items were tested and tallied. Each food sample weighed ten

grams was homogenized in 90 milliliters of buffered peptone water. After serial

dilution of the homogenates, a volume of 0.1 ml dilution was spread out on

solid media and incubated for 24 hours at 35–37 °C. Muller Hinton agar was

used to test for antibiotic susceptibility in isolated species, and SPSS version

20.0 was used to enter and interpret the results. A total of 72 food samples
from street vendors were examined for the presence of bacterial infections. A

total of 63 isolates were found after 44 out of 72 samples tested positive and

19 samples contained two pathogens. The overall mean count of aerobic

bacteria was 6.64 × 104 CFU/g, with a range of 1 × 104–1.86 × 105 CFU/g.

S. aureus (53.96%) is the most common iolate, followed by E. Coli (23.8%),

Enterobacter species (15.87%), and Citrobacter species (6.3%). The

antimicrobials that were shown to be most effective against all isolates were

gentamycin, chloramphenicol, tetracycline, ciprofloxacin, and trimethoprim-

sulfamethoxazole; ampicillin and ceftazidime were found to be the least

effective. (Amare et al., 2019).

A microbiological analysis was performed in the study about

Microbiological assessment of ethnic street foods of the Himalayas in order to

determine the pathogens on the food samples (samosa, kachori, puchkka, alu

chop, vegetable momo, pork momo, alu-cheura, vegetable chowmein, jhal-

muri, sya-faley, samosa, kachori, pani puri, alu tikki, vegetable momo,

mutton momo, bread chop, vegetable chowmein, jhal-muri, and vegetable

pakoda). The study performed Bacillus diarrheal enterotoxin visual

immunoassay and most probable number (MPN) count of coliforms.

Lactococcus lactis, Lactobacillus plantarum, Lactobacillus brevis,

Enterococcus faecium, Bacillus subtilis, Bacillus pumilus, Bacillus

licheniformis, Bacillus cereus, B. cereus, Clostridium perfringens, S. aureus,

Salmonella spp, Escherichia coli, Salmonella spp., and S. aureus were

detected on the food samples (Kharel et al., 2016).

In the study about the detection and count of Escherichia coli in street

foods sauces University of Eastern Philippines, Microbial analysis was

performed on samples that were taken from the street sellers' stalls. In order

to count the colonies in the 3M Petrifilm, the plate was placed in an

improvised colony counter (lamp shade and magnifying lens). This allowed the

researchers to see the colonies clearly and began counting, using the same

technique that was used to count the red and white blood cells in a pattern

resembling a snake. If there are more than 150 colonies on the plate, an

estimate must be made. To do this, count the colonies using a bacterial colony

counter in one or more representative squares. Then, calculate the average

number per square. Finally, multiply the average number by 20 to get the

estimated count per plate. ((Dagalea et al. 2021).

2.3 Related Studies

Food Safety Knowledge of Vendor

Street food is a weak link in food safety supervision and has been the

focus of academic study for a long time. In addition to being convenient for

many, street food offers millions of low-income individuals with a means of

subsistence, greatly boosting the economies of many developing nations. (Ma

et al., 2019). Ethiopian street food is becoming more and more famous in the

coutry. There are currently a lot of street sellers in the area around bus

stations, schools, and hospitals. Customers who purchase food from street

vendors run the danger of catching foodborne illnesses because the food may

be contaminated with various diseases and provided in unsafe conditions near

busy roads. Few studies have been conducted on Ethiopian street food,
therefore nothing is known about its hygienic quality. The local government

doesn't even have a system in place to regulate street food vendors to

guarantee food safety. (Negassa et al., 2023). Street food is an affordable,

ready-to-eat dinner option for those who are poor and underprivileged in

emerging nations. However, due to inadequate vendor cleanliness and the

security of the selling place, the street food industry poses a serious risk to

public health. (Mohammed et al., 2023). Access to nutritious food is seen as a

basic human right, but food-borne illness is a significant global public health

concern. In low- and middle-income nations, the issue is worse. Due to the

rise in urbanization and the appeal of street food in low- and middle-income

countries, it is imperative to comprehend the Knowledge, Attitude, and

Practice (KAP) of street food vendors in order to ensure food safety. (Desye et

al., 2023). People are eating meals outside of their homes more frequently,

which increases the risk to foodborne infections. Unsafe food preparation and

supply by vendors has made food safety a public health concern. The purpose

of the current study was to evaluate the knowledge, attitudes, and food safety

and hygiene practices of street food vendors (SFVs) in Northern Ghana.

(Yakubu et al., 2023).

In the study about determining knowledges, attitudes and practices

(KAP) of street food vendors, within South Africa in the city of Johannesburg A

descriptive study design utilizing a quantitative method to describe the

knowledge, attitudes and practices of street food vendors out of the 150 street

food vendors (SFVs) that took part in the survey. Although most of the

merchants could not enumerate the five keys to safer foods, the merchants

knew enough about the real behaviors associated with each key. Attitudes

toward specific food safety activities were also good, as evidenced by the fact
that all items showed more than 71% agreement. (Campbell et al., 2011). In

another study about evaluation of the street food vendors’ knowledge,

attitudes, and practices regarding food safety as well as the associated factors

that may affect those practices in Mekelle City, Northern Ethiopia, a

community-based cross-sectional study design was employed from February

2020 to August 2020 to assess the level of knowledge, attitude, and practices

of street food vendors and their associated factors with food safety practice. All

of the street food vendors in Mekelle, Ethiopia, are the source population . The

research population consisted of all street food sellers who had been cooking

and selling prepared food on the streets, in temporary improvised structures,

or at home before setting up shop on the streets of Mekelle City. Regarding

food safety, the majority of study participants—117 (65%), 146 (81.1%), and

106 (58.9%)—had good levels of knowledge, a positive attitude, and good levels

of practice. Age (AOR: 0.33, 95% CI: 0.14-0.77), monthly income (AOR: 5.2,

95% CI: 1.42-18.9), educational status (AOR: 0.07, 95% CI: 0.009-0.623),

experience (AOR: 0.54, 95% CI: 0.22-1.34), food safety training (AOR: 0.490,

95% CI: 0.317-0.757), and understanding of food safety are among the factors

that influence street food vendors. (Werkneh et al., 2023). Food handlers in

food establishments had a higher level of compliance with food hygiene and

safety measures than food handlers in street food shops. In the study about

food hygiene and safety measures among food handlers in street food shops

and food establishments of Dessie town, Ethiopia a community-based cross-

sectional study was performed from December 2013 to June 2014. Using a

pre-tested structured questionnaire and an observational checklist, a trained

data collector gathered information from 116 food handlers from 116

restaurants and 19 food handlers from 19 street food trucks. To determine the

parameters substantially related with high standards of food safety and

cleanliness, multivariable logistic regression analysis with a 95% confidence

interval (CI) was employed. Overall compliance was rated as "very good" for

26% of food handlers at restaurants and 16% of those at street food outlets.

Furthermore, a "poor" overall compliance rating was given to 28% of food

handlers in restaurants, compared to 47% of food handlers in street food

vendors. More over three-quarters of restaurant employees (72%) had good

levels of food safety and cleanliness, compared to 53% of those who handled

food in street food outlets. (Adane et al., 2018).

200 street vendors participated in a study about safe handling

procedures and food microbiology in Esprito Santo, Brazil. The purpose of the

study was to assess commercialized foods' microbiological quality and

appropriate handling procedures. The municipalities in Brazil's state of

Esprito Santo were selected according to their financial value and population

size. Vitória and Serra were consequently selected. The process of producing

the food, the amount of handling, the products' exposure to the market, and

their accessibility were taken into consideration while choosing the sample.

Foods with minimal handling requirements, those without specific

temperature requirements for storage (cold or hot), and those without

requiring storage equipment to be cleaned were all excluded. The list of food

items included grilled meat on skewers, hotdogs, savory snacks, coconut

water (which required refrigeration or conditioning equipment), and boiled

corn. From this total, 49.5% (n = 99) had a single employee to manage all

duties, including handling, sales, and site maintenance, and 54.5% (n = 109)

were in Vitória. This kind of business is defined by the lack of differentiation

between distinct processes such as food handling, money handling, hygienic

practices, and waste collection. This might potentially compromise the quality

of food. (Ferrari et al., 2021). In circumstances where merchants fail to wash

their hands before handling and selling (handling of money, garbage, and

utensils). The person touching the food may be to blame for its contamination

because hands are crucial conduits for the passage of organisms from the

feces, nose, and skin to the food. (Amare et al., 2019; Loukieh et al., 2018) .

Out of all respondents, only 23.8% and 20.9% said that the respondents knew

that Salmonella and Staphylococcus are foodborne infections. Nonetheless,

participants claimed to know more about foodborne illnesses; around two

thirds correctly identified bloody diarrhea (67.5%) and typhoid (62.3%) as

being spread by contaminated food. 52.6% of respondents said the respondets

were either unaware of the need for time off work when infected with an

infectious skin disease or thought it was mandatory. While 73.5 percent of

participants did not know that children, pregnant women, and the elderly

were all equally vulnerable to foodborne infections, 68.2% and 71.5% of

participants knew when to use gloves and how to wash their hands,

respectively. 52.3% of respondents met Bloom's cutoff criteria, indicating

intermediate knowledge, and 39.4% met inadequate knowledge. The

knowledge test had a mean score of 8.99 (SD = 4.17, range: 1–18). The

average food safety knowledge scores significantly (p < 0.05) differed by the

participants’ age, marital status, income, residence and work experience (Abid

et al., 2022). Another study conducted by Daniele et al., (2021) investigated

whether improvements in street-food safety can be achieved by providing

information to vendors in the form of training. While there was minimal

change in the observed behavior, the study found that randomly selected

sellers in Kolkata, India had significantly improved their knowledge and

awareness. Two main explanations were provided by the study for these

findings. Firstly, information collection does not automatically make it more

easier for providers to offer safer food options to customers. Second, even
though consumers are willing to pay for perceived cleanliness, consumers find

it difficult to distinguish between contaminated and safe food. The study made

recommendations for how to deal with supply-side constraints and consumer

education. Daniele et al., (2021). In the study about street food handlers' food

safety knowledge, attitudes and self-reported practices and consumers'

perceptions about street food vending in Maseru, Lesotho a descriptive survey

was conducted around the taxi ranks amongst 48 food handlers and 93

consumers using a semi-structured questionnaire for assessing food handler

knowledge, attitudes and practices, open-ended questionnaire for obtaining

consumer perceptions and observation checklist. Regarding their knowledge of

food safety, vendors received a score of 49% out of 11. On the contrary, Addo-

Tham et al. (2020) found that the Ejisu-Juaben Municipality in Ghana had a

mean knowledge score of 78.35 9.33%, which was considered good. The study

found that while 84% of food handlers were knowledgeable about how to

properly clean and sterilize utensils, 100% of food handlers showed good

awareness of general sanitary procedures such routine hand washing.

However, only 3 percent of respondents knew that using detergent to clean

utensils did not guarantee their cleanliness. While 64% of vendor respondents

appeared to be aware that typhoid fever is spread by food, the majority (86%)

of vendor respondents had poor comprehension of foodborne disease

transmission in high risk groups. (Letuka et al., 2021b).

Bacterial Food Assessments in Salmonella Species

In the study of contamination of street food with multidrug-resistant

Salmonella, in Ouagadougou, Burkina Faso sampled 201 street food stalls, in

11 geographic sectors of Ouagadougou, Burkina Faso between June 2017 and

July 2018. In accordance with ISO 6579:2002, the investigation looked for

Salmonella contamination in 201 sandwiches (one from each seller). Following

serotyping and antimicrobial susceptibility tests, full genome sequencing was

performed on all Salmonella isolates to investigate their phylogenetic

relationships and antibiotic resistance factors. Types of Salmonella. were

discovered in 36 (36.6%) of the 201 (17.9%) tests of sandwich samples. There

were a total of 18 of these contaminated sandwiches, of which 18 (or 50%) had

beef mince, 6 (16.7%) fish, 5 (13.9%) beef kilichi, 3 (8.3%) omelettes, and 1

(2.8%) avocado. No species of Salmonella. Sandwiches with sausages included

strains. 16 distinct serotypes of Salmonella were identified after the 36

isolates were antigenically described. Salmonella enterica was present in

36/201 cases, or 17.9% of cases, and the isolates belonged to 16 different

serotypes, with Kentucky, Derby, and Tennessee accounting for the majority

of isolates with five each. Brancaster and Kentucky serotypes were most

frequently seen in six Salmonella isolates. (Nikiema et al., 2021).

Bacterial Food Assessments with other Bacteria species

Making sure there is enough food to feed everyone in a developing

country's population is one of the biggest concerns. (Salamandane et al.,

2021b). Street food has great cultural and economic significance and is

consumed all around the world. In addition to serving quick and reasonably

priced meals to city dwellers, street food vendors, especially women, often rely

on it as a vital source of income. Street food may also be a reflection of the

traditional local culture, which is a big lure for tourists. Despite these

benefits, the fact that street food is sometimes made and sold under unsafe

conditions without government control or hygienic oversight has raised

concerns about its microbiological safety. Consequently, street foods have

been linked to a number of foodborne illnesses as well as high concentrations

of fecal indicator bacteria. (De Andrade et al., 2023). Street Foods also play a

significant part in supplying many urban residents with affordable,

wholesome, and readily available food, but if those foods are made in an

unclean and uncontrolled environment, street foods may increase the burden

on food safety (Birgen et al., 2020). Therefore, the demand to ensure the

supply of safe food has been one of the major challenges and concerns for

producers, consumers and public health officials globally (Moloi et al., 2021).

Although there are many benefits to buying food from a street vendor,

there is evidence that some of the food may be contaminated with dangerous

germs. There is a dearth of information, however, about the bacteriological

profile, bacterial load, and antibiotic susceptibility patterns of bacterial

isolates from street food in Gondar, Northwest Ethiopia. (Amare et al., 2019).

In Hawassa, Ethiopia, researchers looked at the bacterial quality of

street food and the antimicrobial resistance of isolates. A cross-sectional study

was conducted from May to September 2014 A total of 72 samples from six

food items such as local bread (‘ambasha’ and‘kita’), raw fish, chilli (‘awaze’),

avocado and cooked potato were collected. Standard microbiological

techniques were followed in the isolation of bacteria, colony count, and

antimicrobia susceptibility testing. The researchers discovered that the total

colony counts of 31% of the food samples varied from 1.7 105 to 6.7 106

colony-forming units per gram (CFU/g), above the permitted limits for the

microbiological quality of foods that are ready to eat. Raw fish, "kita," and

"ambasha" had mean coliform and enterobacteriaceae levels that were higher

than allowed, in addition to exceeding the limits. E . coli was the most often

occurring isolate (29.6%), followed by Salmonella species (12.7%) and

S. aureus (9.9%). Ciprofloxacin was effective against all isolates in 100% of

cases. Roughly 89% of Salmonella spp. were resistant to chloramphenicol.

14.3% of S. aureus, uncomfortably, were vancomycin resistant. (Eromo et al.,

2016)

A study about the microbiological assessment of Staphylococcus

spp. found that 233 samples of different street foods contained the following

bacteria: Salmonella enteritica, Bacillus subtilis, Bacillus pumilus, Bacillus

licheniformis, Bacillus cereus, Escherichia coli, Enterobacter aerogenes,

Enterobacter cloacae, and Shigella flexneri. and Enterobacteriaceae were

found to be the two most common pollutants. plus B. cereus in food samples

that were analyzed. When evaluated, only a small percentage of street goods

released pollutants. There were no Salmonella or Staphylococcus enterotoxins

found in the meals that were tested (Kharel et al., 2016)

According to another study about microbiological assessment of

street foods at the point of sale in Maputo (Mozambique) From 83 different

vendors selling different types of foods, 83 samples of ready-to-eat (RTE) street

food were analyzed. Escherichia coli, total coliforms, and mesophiles were

used as indicators of quality and hygiene. Listeria monocytogenes (L.

monocytogenes), Salmonella, and coagulase-positive staphylococci were used

as indicators of food safety. Coagulase-positive staphylococci and E. coli, total

coliforms were counted by dispersing 0.1 mL of the corresponding decimal

dilution in accordance with ISO (2001) and ISO (1999a). After a 24-hour

period at 37 ± 1 °C, the amount of E, coli colonies and total coliforms were

counted onto Compass ECC Agar medium (Biokar Diagnostics, Beauvais,

France) (ISO, 2001). Staphylococcus spp. Colonies were tallied on Baird Parker

Agar medium (Biokar Diagnostics, Beauvais, France) after 48 hours at 37 °C

(ISO, 1999a). Then, coagulase testing was carried out in accordance with ISO (1999b) in tubes

holding 0.1 mL of culture and 0.3 mL of rabbit plasma (Biokar Diagnostics, Beauvais, France).
High percentages of poor meal samples were found in both traditional hot

(76.7%) and cold (75%) foods. The results of the study's monocytogenes and

L. salmonella testing were negative. Nevertheless, using coagulase-positive

staphylococci as a food safety indicator, 25% (23/83) of the food samples

analyzed were classified as inadequate or maybe dangerous. Only a few street

foods tested positive for toxin production, and neither Salmonella toxins nor

Staphylococcus enterotoxins were found in the street foods tested. The

predominant contaminant bacteria were enterobacteriaceae, followed by

Staphylococcus spp. and B. cereus.. (Salamandane et al., 2021c).

This study will have the same structure of Salamandane et al., 2021 &

2018, Eromo et al., 2016 as these studies have identified the abundance of

the bacteria species S. aureus, E. coli, Salmonella, which the researchers aim

to achieve.

Based from the studies of Werkneh et al., 2023, Adane et al., 2018, and

Eromo et al., 2016, in order to achieve the desired outcome of this study the

researchers will follow the community-based cross-sectional study design

instead of following the study Campbell et al., 2011, which followed

descriptive study design utilizing a quantitative method.

This study will be using “most probable number (MPN) of coliforms” and

“immunoassay for detection of enterotoxins and toxins of pathogenic bacteria”

in determining the quantitative and qualitative characteristics of the presence

of microorganisms in street foods based from the study of Kharel et al., 2016

as well as Salmonella sp. Detection based from the study of Nikiema et al.,
2021 including Staphylococcus aureus Count and E. coli Count based from the

study of Salamandane et al., 2021.

The review of related literature has helped this study to identify what

kind of research design the researchers will employ in order to achieve its

desired outcome. The review of related literature has also to helped this study

by allowing the researchers to identify the structure the researchers will follow

including the kinds of parameters to be employed in this study to achieve its

desired outcome.

CHAPTER III
METHODOLOGY

3.1 Introduction
This chapter demonstrates the approach used to collect the study's data. It
describes the research design, sample size selection, sampling design,
technique, subject, research site, instrument, instrument validation, data
collection process, data processing approach, and statistical analysis.

3.2 Research Design

Data from one particular point in time are examined in a cross-sectional
study. In this kind of study, participants are chosen based on specific
variables of interest. A community's characteristics can be described using
this kind of research, but cause-and-effect relationships between various
variables cannot be established. This technique is frequently used to draw
conclusions about potential connections or to collect initial data to support
future study and experimentation (MSEd, 2022).

3.4 Research Instrument

The instruments that will be used to achieve this study’s objectives are:

1. Salmonella sp. Detection, Presumptive (Conventional) -

Presumptive detection of Salmonella refers to the initial screening test that
indicates the possible presence of Salmonella in a sample. The conventional
method for detecting Salmonella involves pre-enrichment in buffered peptone
water and enrichment in selective media, followed by isolation on differential
media and serological confirmation (Ferretti et al., 2001). This kind of testing
parameter is required for this research as it will help determine the presence
and abundance of Salmonella present in the street food samples.

2. Staphylococcus aureus Count- Staphylococcus aureus count refers

to the number of Staphylococcus aureus bacteria present in a sample. The
acceptable level of Staphylococcus aureus in food is below 10^3 colony-
forming units per gram (cfu/g) of food. If the amount of bacteria is greater
than 10^4 cfu/g, the food is unsatisfactory and potentially hazardous for
health and/or unfit for human consumption (Tsehayneh et. al., 2021). This
kind of testing parameter will be used in this study as it will help determine
the presence and abundance of Staphylococcus aureus present in the street
food samples.

3. E. coli Count- E. coli count refers to the number of Escherichia coli

bacteria present in a water sample. E. coli is a type of coliform bacteria that is
found in the intestines of warm-blooded animals, including humans, and is
used as an indicator of fecal contamination in water. The presence of E. coli in
water indicates recent fecal contamination and may indicate the possible
presence of disease-causing pathogens (New York State Department of Health,
2023), such as bacteria, viruses, and parasites. The acceptable level of E. coli
in drinking water is determined by risk analysis based on statistics to protect
human health. Drinking water should have no E. coli after treatment. E. coli
levels at designated swimming beaches should not exceed 88 per 100 milliliter
(mL) in any one sample, or exceed a three-sample geometric mean average
over a 60-day period of 47/100 mL (New Hampshire DES, 2019). This testing
parameter will be used to determine the presence and abundance of E. coli in
the street food samples.

4. Most Probable Count (MPN) of coliforms- The Most Probable

Number (MPN) is a statistical method used to estimate the concentration of
viable microorganisms, including coliforms, in a sample. It is commonly used
in the microbiological assessment of water and food, including street food. The
MPN test is particularly useful for samples that contain particulate material
that interferes with other methods of analysis. In the context of street food's
microbiological assessment, the MPN method can be used to estimate the level
of coliform contamination, which is an indicator of fecal contamination and
overall hygiene. High coliform counts in street food can indicate potential
health risks. The MPN test helps in evaluating the bacteriological safety of
street vended food and identifying the relationship with coliform count. It is an
important tool for ensuring the safety and quality of street food products
(Rijal, 2023).

5. Immunoassay- Immunoassays are commonly used for the detection

of enterotoxins and toxins of pathogenic bacteria in street food. These assays
involve the use of specific antibodies to detect the presence of toxins. For
instance, a magnetic bead-based immunoassay was developed for the
simultaneous detection of E. coli, Salmonella spp., and S. aureus in animal-
derived foods (Kim, 2018).

3.5 Validation of the Research Intrument

The Department of Science and Technology (DOST) will oversee and
validate the research instrument's effectiveness and accuracy in detecting
these foodborne pathogens in street food samples. This validation will involve
rigorous testing, comparison with established standards and methods, and
statistical analysis to ensure the instrument's reliability.

The validation process will also consider factors such as precision,

sensitivity, specificity, and repeatability to guarantee that the research
instrument provides consistent and accurate results. DOST will collaborate
with experts in microbiology and food safety to validate the instrument
thoroughly.

Upon successful validation, the research instrument can be confidently

used for assessing the safety of street food, protecting public health, and
ensuring food regulatory compliance. The results obtained through this
instrument will serve as critical data for food safety assessments and
regulatory decision-making processes.

3.6 Data Gathering Procedure

Step 1. Approach the LGU and the City Health Office in order to know
the registered food stalls in the Bayugan City Night market.

Step 2. Ask the LGU, City Health Office and the Baranggay for
permission to conduct the Research within the Bayugan City Night Market.

Step 3. Conduct the research in the Bayugan City Night Market.

Step 4. Buy the street food samples from the registered food stalls only,
3 samples of each kind will be needed.

Step 5. Place the street food samples in sterile and cool containers,

Step 6. Transport the samples to the DOST CARAGA Lab for the testing
of the samples
 Step 7. Have the samples tested in Staphylococcus aureus Count,
Salmonella sp. Detection, Presumptive (Conventional), E. coli Count.

Step 8. Collect the test results from the DOST CARAGA Lab.

3.7 Statistical Analysis

This study will use descriptive statistical tools to analyze the data
collected from the street food samples from the Bayugan City Night Market.
This study will use frequency, mean, and percentage to interpret the the
collected data from the night market of Bayugan City
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