Evaluation of Bacterial Population on Areas with Common Contact

Surfaces

ABSTRACT
Bacterial contamination and infection are currently major health concerns when it comes to humans. Bacteria can
attach themselves to surfaces and can remain infectious for several hours or days. They can transfer through surfaces
and can be transmitted through contact that occurs frequently throughout the school day. The study aimed to (1)
determine the collected sample with the greatest number of colonies (2) identify the presence of gram-positive and
gram-negative bacteria in the collected samples and (3) evaluate the difference between and among all collected
samples. The study, if conducted successfully, will increase awareness of the high-contamination surfaces in the
University of Santo Tomas-Legazpi that people should be wary of. The study will also be of benefit to the students
by pinpointing which surfaces need to be cleaned and monitored. This way, bacterial infection, and contamination
can be reduced. The study utilized an experimental research design to gather data. The researchers used sterile swabs
and performed the swab sampling method. Results showed that the samples from the cafeteria exhibited the greatest
growth in bacterial colonies after 18 hours of incubation. After gram staining was done, 7 out of 19 surfaces had the
presence of gram-positive bacteria, while the remaining 12 surfaces were shown to have the presence of gram-
negative bacteria. Most gram-negative bacteria were found in the bathroom and classroom, while the gram-positive
bacteria were mostly found in the cafeteria. The researchers concluded that given the significant difference in
statistical data, special attention must be given to the cleanliness of seats and tables. In light of the limitations of the
study and other factors that affect the study, the researchers would like to recommend future researchers to further
identify the characteristics of the gram-positive and gram-negative bacteria and identify the type of bacteria.

Keywords: bacterial colonies, contact surfaces, gram-positive, gram-negative, University of Santo Tomas-Legazpi

INTRODUCTION

Currently, due to their way of life, the

specificity of work, and epidemic threats, people
spend most of their time indoors (Jabłońska-Trypuć,
2022). In the way the world works today, humans
spend the majority of their lives indoors, from the
comfort of structures designed for safety and
protection. However, even while we are inside, we
still encounter danger in the forms of microorganisms
that are virtually on every surface we come in contact
with. The frequent exposure we experience to
bacteria carries with it the potential for bacterial
contamination as well as transmission (Meadow et
al., 2014).

School attendance is a recognized risk

factor for infectious diseases among children. One
particular concern is the increased possibility of
antimicrobial-resistant bacteria developing among
these children (Alkhamis & Noweir, 2018). If certain
physical conditions, such as moisture level,
temperature, and the presence of organic and
inorganic substrates, are met in a school
building/environment, and microbes can easily
proliferate. Just as buildings differ from one another
and local government areas differ, so do bacterial
concentrations in their indoor and outdoor
environment (Akinrotoye et al., 2018).
In schools, where frequent interaction with
many germs occurs during the day, microorganisms
pose a severe hazard. Numerous microorganisms
inhabit the human body, but they are all healthy and
only have the potential to be harmful if they shift
where they are located in the body. Various kinds of
microorganisms are easily contacted by the
environment and the hands can be the most important
means by which pathogenic microorganisms are
transmitted (Maji et al., 2018). Bacterial infection
may happen when there are too many pathogens in the
body. When this happens, the bacteria can quickly
multiply and be detrimental to our health. One such
example is the spread of Norovirus, a very contagious
virus that a person can get infected by coming in
contact with contaminated surfaces. These viruses are
responsible for approximately 20% of all cases of
acute gastroenteritis worldwide. They cause an
estimated 200,000 child deaths each year and are now
the most common etiological agent of pediatric
diarrhea in rotavirus-vaccinated populations
(Nordgren, 2019).
According to data from a study published by
Lancet Journals in 2019, it was stated that “Bacterial
infections are the second leading cause of death
worldwide”. They found that from an estimated 13·7
million (95% UI 10·9–17·1) infection-related deaths
in 2019, there were 7·7 million deaths (5·7–10·2)
associated with the 33 bacterial pathogens (both
resistant and susceptible to antimicrobials) across the
11 infectious syndromes were estimated in their study.
They found that only five among the studied
pathogens including S. aureus, E. coli, S.
pneumoniae, K. pneumoniae, and P. aeruginosa
accounted for more than half of all bacteria-related
deaths caused by the pathogens studied.
Bacteria are ubiquitous microorganisms in
both outdoor and indoor environments, identified as
microbial sources of contamination. Inanimate objects
(fomites) have been shown to play a role in the
transmission of human pathogens either directly, by
surface-to-mouth contact, or indirectly, by
contamination of fingers and subsequent hand-to-
mouth contact (Akinrotoye et al., 2018).
The Gram stain easily divides bacteria into
two groups, Gram-positive and Gram-negative,
based on their cell wall and cell membrane
permeability (Thairu et al., 2014). Gram-positive
bacteria have thick cell walls, outside the cell
membrane, and contain other macromolecules
(carbohydrates, lipids, proteins, etc.). Meanwhile,
gram-negative bacteria are thin and overlaid by the
outer membrane. For gram-positive bacteria, when
tested using the gram staining procedure, the strain
of the bacteria becomes purple, whilst, for gram-
negative bacteria, it does not (Doron, 2008).
As far as bacterial contamination goes,
bacteria can spread through a variety of routes,
including direct contact with surfaces, objects, and
humans. The prevalence of bacteria corresponds to
the common and frequent spread and growth of
bacteria. It cannot be avoided but can be stopped.
Direct skin contact, bodily fluid exchange, airborne
particles coming in contact, or touching an infected
surface are the most common ways of bacterial
transmission. Microorganisms on the hands can be
transferred to various surfaces, from which they can
re-infect other people (Jabłońska-Trypuć, 2021).
The issuance of an Amendatory DepEd
Order to DO 34, s. 2022 encourages the
implementation of in-person classes due to the
benefits that it will bring in terms of learning
efficiency amongst the students (Department of
Education [DepEd], 2022). But despite that being the
case, it is undeniable that there are still risks that
come with this type of setup. Human coronaviruses
can remain infectious on inanimate surfaces at room
temperature for up to 9 days. Contamination of
frequent touch surfaces in healthcare settings is a
potential source of viral transmission (Kampf et al.,
2020). As the Covid-19 pandemic is still going on,
regulating public health is of the utmost importance.
Conducting face-to-face classes during a pandemic
also warrants extra precautions to ensure the safety
of those to whom the university will be held
accountable. Because of this, there is a need to assess
whether or not different common contact surfaces
are as clean as they should be with the implemented
health protocols. Additionally, full-capacity face-to-
face classes also mean that the school will have a
larger group of people to monitor. This increase in
population size might pose issues regarding the
maintenance of cleanliness around the campus, as it
would be harder to keep track of all the people
visiting certain areas within the school.
The study can serve as a basis for what
surfaces require thorough attention and act as
background or supporting information for the most
common contact surfaces where bacterial growth is
prevalent.
In a study by El-Kased and Gameledin
(2020), they conducted a study to determine the
prevalence of bacteria in primary schools. It was
particularly done in a highly standard private school
and a public school to study what kind of effects
social classes garner on the cleanliness of the schools.
Samples were taken from the places which the
children mostly touch or in contact with such as
water, canteen, desks, books, door handles, banisters,
taps, toilets, and staff hands. The study revealed the
number of bacteria found in public schools is
significantly higher than their private school
counterparts. It must also be noted that both schools
examined are contaminated with different kinds of
bacteria. These results found that children are at high
risk of infection as they are surrounded by a cram of
microorganisms everywhere. The risk of disease
transmission via surfaces (fomites) involves numerous
factors. Some of these factors include frequency of
site contamination and exposure, level of pathogen
excreted by the host, the likelihood of transfer of the
infectious agent to a susceptible individual, virulence
of the organism, immunocompetence of the persons in
contact, and the practice of control measures (i.e.
disinfectant use and personal hygiene), and other
factors (Akinrotoye et al., 2018).
In agreement with the study done by El-
Kased and Gameledin (2020), in which they stated
that “Pathogenic microorganisms are serious threats in
schools, where contact with many microorganisms
happen regularly during the school day.”, Alkhamis
and Noweir (2018) conducted a study where they
randomly selected 16 primary schools where the
sampling sites were the commonly touched surfaces
swabbed during the school visits. These surfaces
include: toilet taps, toilet washers, washing sinks, and
door handles in bathrooms and classrooms. The
findings of their study reveal that single-hand contact
with a contaminated surface results in a variable
degree of enteric pathogen transfer.
Similar to the study of Bhatta et al. (2018),
“Bacterial contamination of frequently touched
objects in a tertiary care hospital of Pokhara, Nepal:
how safe are our hands?”, the researchers aimed to
determine the bacterial contamination of common
contact surfaces in the University to eliminate the risk
of these potential infections. The findings of the study
highlight the need for improved hand hygiene among
healthcare workers and regular
cleaning/disinfection of sites of frequent public
contact.
In a study done by Worku et al. (2018), they
used gram reaction and colony characteristics as
presumptive identification of bacteria They found
that environmental surfaces in healthcare centers act
as a reservoir for bacteria and can as well serve as
vectors of the bacterial pathogens. In hospitals,
viruses, bacteria, and their spores as well as fungi are
transmitted not only by infected patients but also by
hospital staff, visitors, and inanimate hospital
environments, including surfaces frequently touched
by hands, called “high-touch surfaces”, which
include, among others, door handles, light switches,
surfaces in toilets and in the area where the patient is
in the room (Suleyman, 2018).
In Nigeria, Akinrotoye et al. (2018)
conducted an investigation on the Occurrence of
Pathogenic Bacteria on Public Surfaces within
Community Schools in Abeokuta Environs, Ogun
State. The bacterial isolates from the samples were
morphologically and biochemically identified. The
results of the study revealed contamination of almost
all fomites collected from the schools. The results
found the prevalence of bacteria to be statistically
significant, which included Bacillus subtilis (17.66
%), Bacillus mycoides (16 %), Bacillus megaterium
(12.33 %), Pseudomonas aeruginosa (18 %),
Pseudomonas fluorescens (18 %), Enterobacter sp
(16 %), Escherichia coli (19.67 %), Citrobacter
freundii (10.33 %), Klebsiella oxytoca (16 %),
Staphylococcus aureus (14 %), and Staphylococcus
saprophyticus (7 %).
Most of the existing studies show bacterial
contamination and growth on common contact
surfaces in a hospital setting. While there were
studies conducted in a school setting, the focus was
primarily on common contact surfaces of primary
and elementary students, as they stated that kids
have underdeveloped or bad hygiene habits.
Furthermore, previous studies have chosen their
samples by observations, alongside their
observations and ideas on what common contact
surfaces are.
However, there is a lack of studies focusing
on bacterial growth in a high school setting. Further
investigation of bacterial growth and infection in
High School common contact surfaces is required.
Limited studies of bacterial populations on surfaces
are found in the Philippines. To identify sample
locations, the researchers decided to primarily ask
students of the Senior High School Department of
the
University of Santo Tomas-Legazpi to identify their
most common contact surfaces.
Identifying the growth of bacteria on
common contact surfaces is crucial because bacteria
are a major source of illness in students. That is
especially the case with the recent implementation of
100% full-capacity face-to-face classes. As such,
there is a need for disinfection, proper sanitation, and
overall public cleanliness to be maintained. Finding
the most bacteria-contaminated areas in the University
of Santo Tomas-Legazpi can help reduce illness
because precautions will then be taken to clean these
areas. It is crucial to reduce bacterial contamination or
infection, particularly in light of the ongoing
pandemic. Less bacterial contamination correlates to
less infection and illness among students, allowing
them to attend classes safely and regularly.
The study aims to: (1) determine the
collected sample with the greatest number of colonies
(2) identify the presence of gram-positive and gram-
negative bacteria in the collected samples and
(3) evaluate the difference between and among all
collected samples.
The study, once conducted successfully, will
be beneficial to the following: (1) Students, as the
knowledge gathered can be used to improve their
safety (2) the University, as the data can be used as a
reference for revisions in existing policies that
concern the maintenance of cleanliness (3) and other
researchers, which will be able to use said knowledge
in the field in their future endeavors.
The study focuses on determining the
bacterial growth and population on surfaces
commonly contacted by students in the University of
Santo Tomas-Legazpi Senior High school building.
The study will be conducted within the second
semester of the school year 2022-2023. The study
however does not determine the type of bacteria
present on such surfaces. It is only set on determining
which surface bacteria is prevalent within the campus
and determining if the bacteria present is gram-
positive or gram-negative. Due to the lack of
resources as well as the time allotted for the testing, it
will be delimited to only the building where the
respondents reside and the gathered data may not be
able to precisely provide the bacterial population
within the campus.
METHODS
This chapter defines the research methods
used in the study. It presents the methodology used
for collecting data. It explains the target areas,
sampling technique, collection of samples,
preparation of plates, incubation of samples, and data
analysis.
Research Design
The research followed
a descriptive-experimental-
quantitative design. The researchers planned to go
through data collection by means of experimental
testing of the areas identified. The study is
descriptive as it aims to describe what is going on in
the identified contact surfaces with regard to
bacterial population. The goal is to discern the
parameters regarding the bacterial population.
Materials
Materials used for the laboratory were
experiments, sterile swabs, nutrient agar plates,
crystal violet strain, iodine solutions, and
decolorizer. For testing and swabbing the identified
areas, sterile swabs were used to gather the bacteria
to avoid any outside influence, and the nutrient agar
plate where the bacteria-infested swab will be
inoculated in. It is then used to host the bacteria and
contribute to the growth of said bacteria. The
researchers used 57 of the aforementioned materials
in conducting the testing. The crystal violet strain,
iodine solutions, and decolorizer were utilized in
gram staining.
Target Area
The study’s target areas for testing were the
bathrooms, cafeterias, and classrooms. The areas
were determined as the areas where students come in
contact with high-touch surfaces. Due to many
factors that inanimate surfaces have, such as
moisture and fecal matter, inanimate surfaces allow
the transmission of pathogens and are a source of
growth for bacteria, especially those locations
frequently visited by students (Alkhamis & Noweir,
2018). The research of Alkhamis and Noweir (2018)
stated that toilet washers, sink walls, and door
handles showed positive results for fecal coliforms
due to constant surface interactions with people.
Commonly touched surfaces for sample collection,
including door knobs, desks and chairs of students,
bookshelves, shared toys, and walls were chosen
because the study has categorized these as high-
contact surfaces (Fongt, 2020). On a high-touch
surface, there are more bacteria and more process consisted of suspending and dissolving 28 g
opportunities for the bacteria to process horizontal of nutritious agar powder in 1 liter of purified water.
gene transfer (Lambert, 2022). Bathrooms were also The mixture was stirred while heating to help all the
tested because there are many areas of highly ingredients dissolve completely. The dissolved
potential sources of infection and one of them is mixture was autoclaved for 15 minutes at 121
toilets, flushing rims, and porcelain surfaces due to degrees Celsius. They allowed the nutritional agar to
contamination with microbes from the gut most of cool and did not allow it to solidify once it had been
the time (Alkhamis & Noweir, 2018). autoclaved. Each plate was then filled with nutrient
agar, and the plates were left on the sterile surface
Sampling Technique until the agar was set (Aryal, 2022).
Purposive sampling was used to determine Collection of Samples
the number of surfaces to be tested within an area.
Each selected area was from the results of previous A total of 57 samples were collected from
research that determined which areas have common the surfaces identified as the most commonly
contact surfaces with students. contacted by students in the 3 chosen areas. In each
of the 3 areas, 19 contact surfaces were tested
Procedures wherein 3 samples were collected per surface.
Microbiological Manuals/Protocols
For each of the surfaces, a swab sampling
method will be utilized to collect the samples.
The study follows Public Health England
According to the Monitoring and Sampling Manual
(PHE) standard method for the detection and
of the Department of Environment and Sciences
enumeration of bacteria in swabs and other
(DES) in Queensland, Swab (or wipe) sampling can
environmental samples.
be used to detect organic and inorganic contaminants
on different surfaces. The researchers will utilize a
Aseptic Technique
sterile plastic swabbing template with a standard area
of measurement of 10 cm2. Surface samples are to be
The researchers followed aseptic techniques.
categorized as UST-L campus classroom (3),
They observed the use of lab coats, gloves, masks,
cafeteria (3), and bathroom (3).
hairnets, and other PPE during the research
experiment. Proper health protocols were done before,
during, and after the experiment (Siddiquee, 2017).
Sanders (2012) provided the following aseptic
techniques for plating methods. (1)They were familiar
with all laboratory rules and safety precautions to be
taken when working with microorganisms. (2) They
sterilized all instruments, solutions, and media prior to
using them for plating procedures. (3) They cleared
away all materials cluttering their work area on the
laboratory bench. (4) They cleaned the work area with
disinfectant to minimize possible contamination. (4)
Figure 1. 10x10 Sterile Template
They set up a Bunsen burner and worked slowly,
carefully, and deliberately within the sterile field area
Streaking the Collected Samples in the Nutrient
created by the updraft of the flame. (5) They arranged
Agar Plates
all the supplies needed for the procedure on the
laboratory bench near the sterile field. (6) They
For all samples, dilutions were selected that
washed their hands thoroughly with antiseptic soap
gave colony counts within the appropriate range for
and warm water before handling microorganisms.
the test being performed. (Public Health England)
Nutrient broth served as the liquid medium for
Preparing Nutrient Agar
dilution. The diluted swabs were inoculated to the
nutrient agar in a “wave-like” pattern. The plate was
Nutrient agar/broth was used for the
then rotated 90 degrees and the pattern was repeated.
cultivation and maintenance of non-fastidious
When they introduced the bacteria to the agar, they
organisms and the enumeration of organisms in water,
then sealed, labeled, and stored the plate in a cold
sewage, dairy products, feces, and other materials
place.
(Tankeshwar, 2022). The preparation

Incubation of the Nutrient Agar Plates
The inoculated plates were placed inside a
zip-lock bag to prevent them from drying out and to
control odors. Plates were incubated upside down
(agar up), at 35 degrees Celsius over the course of 18-
24 hours. The plates were then collected and
examined for bacterial growth.
solution:
often the mixed solvent of ethanol and acetone. The
slide was rinsed with water in 5 seconds. The smear
was counterstained with basic fuchsin solution for 40
to 60 seconds. The fuchsin solution was washed off
with water, and excess water was blotted with the
bibulous paper (Tripathi et al., 2022).
Ethical Considerations

Statistical Analysis The researchers abided by the principles of

In order to measure the bacterial growth, the
researchers used a colony counting machine that was
provided by the school. They used dilution in order to
get the right concentration. Serial dilution is the
simplest technique for obtaining manageable
concentrations of a desired organism and it is
complemented by petri dish streaking and spreading,
just two of many plating techniques used by
microbiologists (Blaize et al., 1993). They used the
following formula to get the fixed amount of dilute
This chapter presents
Where: the
discussions, and
C₁V₁ = C₂V₂
C₁ = concentration of the starting solution. C₂
=concentration of the final solution.
V₁ = volume of the starting
solution V₂ = volume of the final
solution
Serial Dilution
Serial dilution is a series of sequential
dilutions performed to convert a dense solution into a
more usable concentration. To perform a serial
dilution, a sample is taken and diluted through a series
research ethics. The researchers requested
permission to swab the identified areas and occupy
and use the university laboratory for experiments.
The researchers then requested for materials to be
used and tools to be borrowed for the entirety of the
experiment. The researchers ensure that protocols
and aseptic techniques are followed during the
experiment to prevent health risks and cross-
contamination.
RESULTS AND DISCUSSIONS
results,
recommendations that
the
researchers have gathered through the process of
experimentation of identifying the amount of colony-
forming units in common contact surfaces in the
University of Santo Tomas-Legazpi St. Albert
Building.
Table 1. Amount of Colonies per Surface Sample
in the Bathroom Area
Dilution
Bathroom Factor
1 1:10 1:100

of standard volumes of sterile diluent, which can Bathroom Flush TNTC TNTC 1092 Colonies
either be distilled water or 0.9 % saline. The extent of 1

dilution is determined depending on the estimated Bathroom Flush TNTC TNTC 1320 Colonies
2
concentration of cells/organisms in a sample. (Society
for General Microbiology, 2016) Serial dilution tests Bathroom Seat 1 TNTC TNTC TNTC

measure the concentration of a target microbe in a Bathroom Seat 2 TNTC TNTC TNTC
sample with an estimate called the most probable
number (MPN) (Blodgett, 2000). Bathroom Faucet
1
TNTC TNTC 1684 Colonies

Bathroom Faucet TNTC TNTC 1985 Colonies

Gram Staining 2

Bathroom Sink 1 TNTC TNTC 1724 Colonies

The researchers used gram staining to make
presumptive identification of bacteria. The testing Bathroom Sink 2 TNTC TNTC 1248 Colonies

began with crystal violet stains added over the fixed

culture. After 10 to 60 seconds, the stain is poured off,
and the excess stain is rinsed with water. Iodine
solution was used to cover the smear for 10 to 60
seconds. The iodine solution was poured off, and the
slide was rinsed with running water. A few drops of
decolorizer were added to the slide. Decolorizers are
Table 1 shows the samples with dilution
factors of 1 and 1:10 exhibited colony counts that
were too numerous to count (TNTC). The number of
colonies that formed in each sample per surface with
a dilution factor of 1:100 in the
bathroom area revealed that the
bathroom seat samples exhibited the
most bacterial growth after 18 hours
of incubation. The bathroom faucet
samples exhibited the next highest
bacterial growth. The bathroom sink
samples while the bathroom flush
samples exhibited the least amount of
bacterial growth. The number of Table 3 shows the samples with dilution
colony forming units determine the
number of active organisms on
common contact surfaces in the
bathroom area. Higher colony count
suggests a dirtier
area than those with lower colony counts. exhibited a higher count of colonies formed than
both the cafeteria table samples.The number of
Table 2. Amount of Colonies per Surface Sample colony forming units determine the number of active
in the Classroom Area organisms on common contact surfaces in the
bathroom area. Higher colony count suggests a
Classroom
Dilution
Factor
dirtier area than those with lower colony counts.
1 1:10 1:100
Figure 2. Colony Forming Units in bathroom sink
Armchair Desk 1 TNTC TNTC 1940 Colonies sample 2 plate
Armchair Desk 2 TNTC TNTC 1804 Colonies

Armchair Seat 1 TNTC TNTC TNTC

Armchair Seat 2 TNTC TNTC TNTC

Door Knob 1 TNTC TNTC 1360 Colonies

Door Knob 2 TNTC TNTC 1348 Colonies

Light Switch 1 TNTC TNTC 1536 Colonies

Table 2 shows the samples with dilution

factors of 1 and 1:10 exhibited colony counts that
were too numerous to count (TNTC). The number of
colonies that formed in each sample per surface with a
dilution factor of 1:100 in the classroom area revealed
that the armchair seat samples have the highest
amount of bacteria colonies whilst the armchair desk
samples have the second highest. The doorknob
samples hold the lowest amount of bacteria colonies Cafeteria 1 1:10 1:100

after the light switch samples. The number of colony Cafeteria Table 1 TNTC TNTC 2504 Colonies
forming units determines the number of active
organisms on common contact surfaces in the Cafeteria Table 2 TNTC TNTC 2936 Colonies

bathroom area. Higher colony count suggests a dirtier Cafeteria Seat 1 TNTC TNTC 1948 Colonies
area than those with lower colony counts.
Cafeteria Seat 2 TNTC TNTC TNTC

Table 3. Amount of Colonies per Surface Sample

in the Cafeteria Area
factors of 1 and 1:10 exhibited colony counts that
were too numerous to count (TNTC). The amount of
colonies that formed in each sample per surface with
a dilution factor of 1:100 in the cafeteria area shows
the 2nd sample from the cafeteria seat sample
Figure 2 shows the number of bacterial colonies that
formed in the nutrient agar plate from the bathroom
sink sample 2.
Figure 3. Counting of colonies of bathroom faucet
sample 1 plate using a manual colony counter
D
 Door Knob 1 ✓
Door Knob 2 ✓
Light Switch
1
✓
Cafeteria
Table 1
✓
Cafeteria
Table 2
✓
Cafeteria Seat
1
✓
Cafeteria Seat
2
✓
Gram-negative contains an outer lipid membrane
that protects them from outside variables making
them harder to treat. 7 out of the 19 samples
collected were determined to be gram-positive while
the remaining 12 are determined to be gram-
Figure 3 shows the count of colonies in the plate of negative.
bathroom faucet sample 1 using a manual colony
counting machine. Figure 4. Gram staining result of bathroom flush
sample number 1
TABLE 4. Gram Staining Results of Samples per
Surface in Each Area of Testing.

Surfa GRAM STAIN

ce
Sam Positive Negativ
ple e
Bathroom Flush 1 ✓
Bathroom Flush 2 ✓
Bathroom Seat 1 ✓
Bathroom Seat 2 ✓
Bathroom Faucet 1 ✓
Bathroom Faucet 2 ✓
Bathroom Sink 1 ✓
Bathroom Sink 2 ✓ Figure 4 shows the gram staining results of
bathroom flush sample number 1. It distinctively
Armchair Desk 1 ✓
shows that it has a pinkish-red color which indicates
it is gram-negative.
Armchair Desk 2 ✓
Armchair Seat 1 ✓ According to Bright et al. (2009), The
entrance and exit door knobs contain the least
Armchair Seat 2 ✓ amount

Table 4 shows the gram stain result of each

surface sample tested. Gram-positive bacteria
commonly vary and require specific antibiotics.
of bacteria colonies because they aren’t touched
frequently since they are often propped open most of
the day. They also stated that sink faucet handles are
used by multiple students and were also among the
most contaminated classroom surfaces.
Moreover, according to another study titled
“Prevalence of Bacteria in Primary Schools” by El-
Kased and Gameledin (2020), the private school sites
with the highest different bacteria counts recorded
were in the toilet door, toilet flush tap, water,
banisters, and canteen in descending order while the
least sites were the staff hands, books, desks, and
classroom doors in the same order.
In summary, the results of this study were
supported by the study conducted by El-Kased and
Gameledin in 2020 regarding the Prevalence of
Bacteria in Primary Schools. Moreover, it reveals that
common contact areas by students that were not
subjected to frequent cleaning displayed more
bacterial growth than those that were cleaned
regularly.
Conclusion and Recommendations
According to the swab sampling method, the
samples taken from the common contact surfaces in
the cafeteria exhibited the greatest amount of growth
in terms of the number of bacterial colonies.
Furthermore, the sample with the highest amount of
bacterial colonies formed out of all the samples
collected was one of the seats swabbed from the
cafeteria. Overall, nearly all the seat samples taken
showed high bacterial colony count, most being too
numerous to count (TNTC).
Through the use of gram staining, it was
determined that 7 surfaces had the presence of gram-
positive bacteria, while the remaining 12 surfaces
were shown to have the presence of gram-negative
bacteria. For the bathroom area, 6 out of 8 samples
taken were gram-negative bacteria. For the classroom
area, 5 out of 7 samples taken were negative bacteria.
Lastly, for the cafeteria area, 3 out of 4 samples taken
had gram-positive bacteria. Taking into consideration
these findings, it was determined that more gram-
negative bacteria can be found on the common contact
surfaces of bathrooms and classrooms, while more
gram-positive bacteria can be found on the common
contact surfaces of cafeterias.
In conclusion, the results show the cafeteria
area exhibits the highest count of bacterial colonies
among all the areas tested. It was determined that 7
surfaces had the presence of gram-positive bacteria,
while the remaining 12 surfaces were shown to have
the presence of gram-negative bacteria. The results
of the study generally showed there is a significant
difference between the amount of bacteria present on
each surface. Given the significant difference in
statistical data, the researchers concluded that special
attention must be given to the cleanliness of seats
and tables.
In light of the limitations of the study and
other factors that affect the study, the researchers
would like to recommend the following for future
researchers: (1) increase the dilution factor solution,
it will help in counting the bacterial colonies and
make data easier to manage and identify (2) make
use of an automatic colony counter, it will make the
process of gathering data much easier (3) collect
swab samples within a 10 x 10 cm area (4) carefully
apply bacterial swab samples on the agar Petri dishes
in a wavelike pattern, and lastly (5) identify further
the characteristics of the gram-positive and gram-
negative bacteria and identify the type of bacteria.
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