Capstone 2021 Edited Ver.

Antimicrobial Effect of Allium sativum (Garlic) and
Allium cepa (Onion) Tubers Ethanolic Extract

Against Staphylococcus aureus
Senior High Thesis Proposal Presented to the

Faculty of Senior High School Department
In DMMC Institute of Health Science
Tanauan City, Batangas
In Partial Fulfillment of this Requirements

for Senior High School
12-Curie
Jill Ivy A. Diamaro
Alexandra Almine
Cherryl Njel Dimayuga
Carol Galicia
Jignet Liann Reintegrado
2021
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DEDICATION
With our deepest love and gratitude, we humbly dedicate this study to:
The researchers’ beloved family
The researchers’ thesis adviser and research 2 teacher Jon Paul Reyes
The researchers’ Capstone teacher Michelle Opulencia
The researchers’ statistician and class adviser Wennie Rose Matira
The researchers’ friends
The DMMC Institute of Health Sciences
The future researchers
And lastly, to the Almighty God.
DMMC INSTITUTE OF HEALTH AND SCIENCES

#143 Narra St., Mt. view Subd., Brgy. 3, Tanauan City, Batangas
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ACKNOWLEDGEMENT
The researchers are sincerely thankful and would like to record the deepest
appreciation to the following people who made the completion and accomplishment of
this thesis possible:
To Sir Jon Paul Reyes, the researchers’ thesis and research 2 adviser, for
assisting and guiding the researchers throughout the conduct of the study, bestowing
strong support in the experiment, for the encouragement to pursue this study from the
start, for sharing his knowledge and imparting such inspiration to continue and complete
this study.
To Ms. Michelle Opulencia, the researchers’ Capstone teacher, for consistent
support and guidance in accomplishing this study.
To Ms. Wennie Rose Matira, the researchers’ statistician and class adviser,
for lending her busy schedule and giving an effort to guide the researchers and interpret
the outcome of the experiment.
And most importantly, to the almighty God, for blessings, knowledge,
guidance, protection and inner strength the researchers needed to accomplish the research
study.

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Table of Contents
DEDICATION...................................................................................................................................2
Chapter I.......................................................................................................................................4
THE PROBLEM AND ITS BACKGROUND............................................................................4
Chapter I
THE PROBLEM AND ITS BACKGROUND

Onion and Garlic have many benefits and its uses were known especially in cooking
and medical concern. The researchers in this study will explore the credibility of onion and
garlic in an experimental way.
Throughout history, many different cultures have recognized the potential use of
(Allium sativum) garlic for prevention and treatment of different diseases. Recent studies
support the effects of garlic and its extracts in a wide range of applications. These studies
raised the possibility of revival of garlic therapeutic values in different diseases. Different
compounds in garlic are thought to reduce the risk for cardiovascular diseases, have anti-
tumor and anti-microbial effects, and show benefit on high blood glucose concentration.
However, the exact mechanism of all ingredients and their long-term effects are not fully
understood. Further studies are needed to elucidate the pathophysiological mechanisms of
action of garlic as well as its efficacy and safety in treatment of various diseases (Phytomed,
2014).

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Homeopathic remedies are made from plants, but they also come from some of the
foods we eat. Perhaps the most well-known, food-based homeopathic remedy is made from
the common red onion and is called Allium cepa.
The onion, also called the common onion or bulb onion, is the most cultivated species
of the Allium genus, and it is related to the leek, garlic, and shallot. Like its other Allium
family members, onions contain important health-promoting sulfur phytonutrients, such as
allyl propyl sulfoxides. As a result, eating onions help boost immunity, treat infections, lower
cholesterol, reduce blood pressure, reduce the risk of heart disease, and maintain blood sugar
levels.
On the other hand, the homeopathic onion, Allium cepa, is used for a wide variety of
infections, cold symptoms, allergies, eye irritations, and neuralgic pains, along with its many
health benefits and uses (Yaneff, 2017). The antimicrobial activity of Allium species has long
been recognized, with allicin, other thiosulfinates, and their transformation products having
antimicrobial activity. Alliums are inhibitory against all tested microorganisms such as
bacteria, fungi, viruses, and parasites. Alliums inhibit multi-drug-resistant microorganisms
and often work synergistically with common antimicrobials (Kyung, 2012).
Plants belonging to the genus Allium are widely cultivated and used all over the world
as food and medicinal plants. Since ancient times, these plants, particularly garlic (Allium
sativum L.) and onion (Allium cepa L.), have represented important components of typical
recipes and traditional healing systems. Not the least of which, their use as food
biopreservatives is well documented, due to the relevant antibacterial activity of their extracts

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and essential oils. In this study the antimicrobial effect of garlic and onion will be explored
because as time goes by the price of medicine is increasing so it means we will be needing
alternative medicines so that other people who can’t afford medicines in pharmacy will be
given chance to be cured. This will be a lot help to those people who live in rural places and
far from towns.
Purpose statement
The purpose of this study is to use the two Allium Species which are Allium cepa
(Onion) and Allium sativum (Garlic) as an alternative medicine like previous studies but to
use its antimicrobial effect against a microorganism called Staphylococcus aureus. The study
will be helpful in the researchers who will soon use this as a guide in upcoming related
experiments and researches in college like medical technology.
Statement of the Problem
“Antimicrobial Effect of Allium sativum (Garlic) and Allium cepa (Onion) Tubers
Ethanolic Extract against Staphylococcus aureus.
This study was conducted to determine the antimicrobial effect of Allium sativum
(Garlic) and Allium cepa (Onion) Tubers Ethanolic Extract against Staphylococcus aureus.
The following questions need to be answered:
1. Is there any significant difference on the antimicrobial effect of Allium sativum
(Garlic) and Allium cepa (Onion) tubers ethanolic extract against Staphylococcus
aureus using the following concentrations?

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a. 100%
b. 75%
c. 50%
d. 25%
(Garlic) and Allium cepa (Onion) tubers ethanolic extract against Staphylococcus
aureus to positive control?
3. Which concentration of Allium sativum (Garlic) and Allium cepa (Onion) tubers
ethanolic extract has the most antimicrobial effect against Staphylococcus
aureus?
4. How may the findings of the study become helpful in the recovery of wound
infections?
NULL HYPOTHESIS
There is no significant difference on the antimicrobial effects of of Allium sativum
(Garlic) and Allium cepa (Onion) tubers ethanolic extract against Staphylococcus aureus and
positive control.
REVIEW OF RELATED LITERATURE
This section provides an overall picture of what have been written or studied
about the subject under study and the contribution given to the researcher.
FOREIGN STUDIES

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. Nowadays, studies are being conducted by many researchers to discover new
scientific uses of many natural sources. The garlic and onion which are allium species can
have many potential scientific uses.
The benefits of garlic (Allium sativum).
It is scientifically proven that garlic is effectively used in cardiovascular diseases as a
regulator of blood pressure, with dropper effects on glycaemia and high blood cholesterol,
against bacterial, viral, mycotic and parasitic infections (Gongacul, 2010). It's also known that
garlic is a wonderful plant having the properties of empowering immune system, anti-tumour
and antioxidant effects. In this article, according to Gurbuz (2017) the summary of properties
of garlic and its use against bacterial diseases is given. This article is a short review of recent
patents on antimicrobial effect of garlic.
The crude extracts of garlic and ginger applied singly and in combination did not exhibit
any in vitro inhibition on the growth of test organisms. However with lime they inhibited
Bacillus spp, Staphylococcus spp. Salmonella was resistant to most of the decoctions
(Onyeagba, 2014).
Other uses of Allium sativum.
This work reports the antimicrobial effect of garlic (Allium sativum Linn.), ginger
(Zingiber officinale Roscoe) and lime (Citrus aurantifolia Linn.) juice and their various
combinations on some bacterial species. This is in pursuance of the efforts to search for drugs
from plants and the verification of the scientific basis of some known practices in traditional

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medicine (Ugbogu, 2014). Tuberculosis is one of the major public health problems faced
globally.
Resistance of Mycobacterium tuberculosis to antitubercular agents has called for an
urgent need to investigate newer drugs to combat tuberculosis. Garlic (Allium sativum) is an
edible plant which has generated a lot of curiosity throughout human history as a medicinal
plant (Phadatare, 2011). Garlic (Allium sativum L.) is one of the most important species of
the genus Allium and recognized as economically important vegetable throughout the world,
especially around the Mediterranean basin where it is considered as main agricultural product
(Leonardi, 2013).
Health discoveries of Allium sativum.
It is also of great importance because of its therapeutic properties and health-related
benefits against various kinds of diseases such as aches, deafness, diarrhea, constipation,
tumors, and respiratory problems. Health benefits from Allium species, especially garlic, have
been used for centuries to treat various kinds of disorders, and still, there is need of research
to explore its health-related potential (Hamidi, 2015). It is a historic medicinal plant,
originated from central Asia about 6000 years ago, and had been started to use as medicine in
India since 5000 years ago and 3000 years ago in China (Agarwal, 2016). Volatile sulfur
compounds, especially thiosulfates, responsible for pungent aroma, are the main compounds
responsible for its physiological effects. Because of its health benefits, garlic is usually
recommended as dietary supplement (Martins, 2016).

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During the past few decades, antimicrobial resistance has become one of the most
serious and challenging threat for the prevention and treatment of the infectious diseases
(Cars, 2014). Recent advances in research have revealed that there are several natural
products with the potential to eliminate or alleviate several serious human diseases, especially
cardiovascular, neurodegeneration, cancer, and several other important diseases (Vasanthi,
2012). A large number of researches have elaborated several herbs with the ability to produce
antimicrobial compounds as their defense response against the number of different stresses
including microbes (Bednareck, 2012).
Antimicrobial potential of Allium species.
A number of Allium species have antimicrobial potential against variety of microbes
including fungi, bacteria, viruses, and other parasites. Among all the Allium species, garlic is
considered most for antimicrobial research after onion (Kyung, 2012).
Antibacterial potential.
Allium extracts containing thiosulfinates have the potential to retard the growth of Gram-
positive and Gram-negative bacteria. It is, however, reported that garlic can inhibit the Gram-
negative bacteria more than Gram-positive bacteria (Perry, 2010).
Extracts from the garlic are reported to exhibit the effective results against saprobic and
pathogenic bacteria, which are resistant to various drugs (Dubey, 2012).
The study about Allium cepa.

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In the present research program, cost effective and environment friendly gold
nanoparticless were synthesized using the onion (Allium cepa) extract as the reducing agent
(Parida, 2011). The aim of this study was to evaluate the biological and chemical response of
Allium cepa L. exposed to inorganic selenium compounds. Besides the investigation of the
total content of selenium as well as its chemical speciation, the Allium test was used to
evaluate the growth of onion roots and mitotic activity in the roots’ meristem (Kurek, 2014).
In a research works also discovered that crude juices of onion and garlic bulbs exert
inhibition on the growth of E. coli, Pseudomonas pyocyaneus, Salmonella typhi, Bacillus
subtilis, in vitro (Lampe, 2013). Allium is the largest and important representative genus of
the Liliaceae family comprises 450 species. Onion (Allium cepa) is a bulbous plant widely
cultivated in almost every country of the world (Lampe, 2013). The present study investigates
an antimicrobial activities of Allium cepa (onion) extract against tested bacteria (Vibrio,
2010)
(Staphylococcus aureus). The antibacterial activities of the methanolic suspension
shows high effect at all concentration in compression with the aqueous suspension examined
at this study (Vibrio, 2010).
Other benefits and uses of Allium cepa.
Essential oil extraction from the stalk, fresh bulb and mature bulb of allium cepa was
carried out through hydro-distillation using Linkersson apparatus (Belitz, 2014). Antifungal
activity of all the essential oil was carried out against aspergilus niger, aspergilus oryzae,
fusarium oxysporium, aspergilus ficuum and penicillium while antibacterial activity was

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carried out against staphylococcus aureus, bacillus subtilis, escherichia coli, salmonella
typhimurium and enterobacter aerogenes (Kirilov, 2014).
Essential oils of mature, fresh green bulb and fresh green stalk of allium cepa was
extracted through hydro-distillation using Linkersson apparatus. Antibacterial and antifungal
activities of all the essential oils were carried out by disc diffusion method against
fungul srains (aspergilus niger, aspergilus oryzae, fusarium oxysporium, aspergilus ficuum
and penicillium) and bacterial strains (staphylococcus aureus, bacillus subtilis, escherichia
coli, salmonella typhimurium and enterobacter aerogenes). The results revealed that mature
bulb essential oil have maximum zone of inhibition against staphylococcus aureus (12.6 mm),
bacillus subtili (Wani, 2011).
Antibacterial against Staphylococcus aureus / Bulbs.
Various concentrations of methanol and aqueous suspensions of dried Allium cepa bulbs
was screened against gram-positive Staphylococcus aureus. All suspensions showed
inhibitory activity; the highest zone of inhibition was noted with the highest concentration of
methanolic suspension (Vamshi, 2010).
The inhibitory effect of the extract of tasted plant extract against pathogenic bacterial
strain (Staphylococcus aureus ) can introduce the plants as a potential candidate for drug
development for the treatment of ailments caused by this pathogenic (Park,2013).
Onion inhibits platelet aggregation in vitro (Ali et al., 1999; Lawson et al., 1992) and
several platelet inhibitors have been isolated and characterized from onions.

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Staphylococcus aureus is a Gram-positive, round-shaped bacterium that is a member of
the Firmicutes, and it is a usual member of the microbiota of the body, frequently found in the
upper respiratory tract and on the skin. It is often positive for catalase and nitrate reduction
and is a facultative anaerobe that can grow without the need for oxygen.Although S. aureus
usually acts as a commensal of the human microbiota it can also become an opportunistic
pathogen, being a common cause of skin infections including abscesses, respiratory infections
such as sinusitis, and food poisoning (Kluytmans, 2015).
The Allium species facts.
It is concluded from this study that Allium cepa extract has antimicrobial activity against
Staphylococcus aureus. It is expected that using natural products as therapeutic agents will
probably not elicit resistance in microorganisms. It is essential that research should continue
to isolate and purify the active components of this natural herb and use in experimental
animals (Augusti, 2010).
Plants as a source of medicinal compound have continued to play a vital role in the
maintenance of human health since ancient times. One such botanical is garlic. Garlic (Allium
sativum L.) belongs to the Liliaceae family and falls within the group of onion. The bulb and
cloves of the garlic are the most commonly used parts of the plants for medicinal purpose. It
has a typical pungent odor, and antibacterial activity depends on the sulfur-containing
compounds, the major one being allicin, which is produced by enzymatic (alliin lyase)
hydrolysis of allicin after cutting and crushing of the cloves (Elmer, 2018). In previous
studies, allicin exhibits its antimicrobial activity mainly by immediate and total inhibition of

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RNA synthesis, although DNA and protein syntheses are also partially inhibited, suggesting
that RNA is the primary target of allicin action.
The structural differences of the bacterial strains may also play a role in the bacterial
susceptibility to garlic constituents. Apart from antimicrobial action, garlic is found to have
antifungal activity (Davis, 2013).
LOCAL STUDIES
Garlic as an antibiotic.
Garlic is an anti -bacterial agent that can actually inhibits growth of infectious agents and
at the same time protect the body from the pathogens. It is known that the most sensitive
bacterium to garlic is the deadly Bacillus anthracis which causes the diseases anthrax. Even
the forefather of antibiotic medicine Louis Pasture acknowledged garlic to be an effective
antibiotic. Some year’s later garlic was shown to have similar effect/activity as penicillin
(Dikasso, 2019). Later studies should similar activity to modern antibiotic including
Chloramphenicol. Even the blood of garlic eaters can kill bacteria and it is also reported that
the vapor from freshly cut garlic can kill bacteria at a distance of 20 cm! The other, the
common and apparently returning diseases tuberculosis was treated with garlic very
successfully as invading Mycobacterium tuberculosis is sensitive to several of the sulphur
components found in Garlic (Debella, 2014).
Garlic (Allium sativum Linn. of Family Liliaceae, Class Liliopsida) is one of those plants
that had been heavily investigated quite a time (Jabar and Al-Mossawi, 2017; Cavallito and

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Bailey, 2017) and one that has already reached the market together with ginger. It has been
medicinally used for centuries to fight infections (Onyeagba et al., 2006; Abubakar, 2019a).
According to Abubakar (2019a) Jabar and Al-Mossawi (2017), the early Egyptians used it to
treat diarrhea, the ancient Greeks used it to treat intestinal and extra-intestinal diseases, while
the ancient Japanese and Chinese used it to treat headache, flu, sore throat and fever.
Allium cepa as a traditional medicine.
A. cepa is also used traditionally for its medicinal virtues in a plethora of indigenous
cultures. Several publications have been produced in an endeavor to validate such traditional
claims. Nonetheless, there is still a dearth of up-to-date, detailed compilation, and critical
analysis of the traditional and ethnopharmacological propensities of A. cepa. The present
review, therefore, aims to systematically review published literature on the traditional uses,
pharmacological properties, and phytochemical composition of A. cepa. A. cepa was found to
possess a panoply of bioactive compounds and numerous pharmacological properties,
including antimicrobial, antioxidant, analgesic, anti-inflammatory, anti-diabetic,
hypolipidemic, anti-hypertensive, and immunoprotective effects.
Significance of the study
The study aims to benefit the following populations:
To the researchers, this study will give a great help to improve skills and to apply every new
knowledge in real life situations.

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To the health of the people, this will give them idea to use garlic and onion as an alternative
medicine whenever they cannot afford medicine.
To the students, this will help them how to cure their wounds and to determine allium
species antimicrobial effect.
To the future researchers, this will help them to explore more and have more knowledge to
enable them to achieve and successful finished their study.
Scopes and Limitations
The study that was conducted focused itself in the students of DMMCIHS that have
wounds. This study tries to discover the effects of Allium cepa (onion) and Allium sativum
(garlic) and explore their antimicrobial effects through experimentation. The Allium cepa and
Allium sativum was used against Staphylococcus aureus. The result will be helpful and will be
applicable in real life situations.
However, the study was only limited for those selected respondents who have wounds
inside the campus of DMMCIHS and will only help the students in the recovery of the
infected wounds. The study used garlic and onion which are organic. As the study shows
result of such experiments using concentrations to compare which is the best result and to
determine the effects.
Definition of terms
The following terms are listed for a better understanding in the study:

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Allium - a bulbous plant of a genus that includes the onion and its relatives (e.g. garlic, leek,
and chives)
Allium species - The generic name Allium is the Latin word for garlic, and the type species
for the genus is Allium sativum which means "cultivated garlic"
Antimicrobial – an agent that kills microorganisms or stops their growth
Concentrations - the relative amount of a given substance contained within a solution or in a
particular volume of space; the amount of solute per unit volume of solution
Effect – something that inevitably follows an antecedent (such as a cause or agent)
Ethanolic - is a chemical compound, a simple alcohol with the chemical formula C 2H 6O
Extract – remove or take out, especially by effort or force
Infections - the presence of a virus in, or its introduction into, a computer system
Isolated – having minimal contact or little in common with others
Microorganisms - is an organism which is microscopic, which means too small to be seen by
the unaided human eye. It includes bacteria, fungi, archaea, and viruses.
Staphylococcus aureus - is a type of bacteria. It stains Gram positive and is non-moving
small round shaped or non-motile cocci. It is found in grape-like (staphylo-) clusters.
Tubers - a much thickened underground part of a stem or rhizome, e.g. in the potato, serving
as a food reserve and bearing buds from which new plants arise.

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Wounds - an injury to living tissue caused by a cut, blow, or other impact, typically one in
which the skin is cut or broken.
Theoretical Framework
Wound Infection Theory
Focal infection theory is the historical concept that many chronic diseases, including
systemic and common ones, are caused by focal infections. In present medical consensus, a
focal infection is a localized infection, often asymptomatic, that causes disease elsewhere in
the host, but focal infections are fairly infrequent and limited to fairly uncommon diseases
(Yomamoto, 2015). (Distant injury is focal infection's key principle, whereas in ordinary
infectious disease, the infection itself is systemic, as in measles, or the initially infected site is
readily identifiable, and invasion progresses contiguously, as in gangrene.)
Focal infection theory, rather, so explained virtually all diseases, including arthritis,
atherosclerosis, cancer, and mental illnesses. An ancient concept that took modern form
around 1900, focal infection theory was widely accepted in medicine by the 1920s. In the
theory, the focus of infection might lead to secondary infections at sites particularly

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susceptible to such microbial species or toxin. Commonly alleged foci were diverse—
appendix, urinary bladder, gall bladder, kidney, liver, prostate, and nasal sinuses but most
commonly were oral (Babu,2011).
. Besides dental decay and infected tonsils, both dental restorations and especially
endodontically treated teeth were blamed as foci. The putative oral sepsis was countered by
tonsillectomies and tooth extractions, including of endodontically treated teeth and even of
apparently healthy teeth, newly popular approaches—sometimes leaving individuals toothless
—to treat or prevent diverse diseases (Skaug,2010).
Conceptual Framework
This model shows the summary of this study. It can be expressed through the
following framework format:
The IPO Model

INPUT
PROCESS OUTPUT
Combination of allium
sativum and allium - Application of Antimicrobial Effect of
cepa tubers ethanolic minced onion and allium sativum (garlic)
garlic
extract used against and allium sativum
- Analysis of
the staphylococcus cepa (onion) against
statistical tool
aureus isolated from - The assessment of Staphylococcus
wounds and as an results aureus that can be
alternative antibiotic
isolated from wounds.
for wound recovery.

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Figure 1. IPO Model of the study
The study uses input-process-output (IPO) model, or input-process –output
pattern and it is a widely used approach in system analysis. The model was the guide of the
study to easily identify the antimicrobial effect of Allium cepa and Allium sativum against
Staphylococcus aureus. The researchers substituted the allium species. This shows
information that will be helpful in understanding the study.
Chapter II
METHODS OF RESEARCH
This chapter of the study shows the method of the extraction of Allium cepa (onion)
and Allium Sativum (garlic) and identification of presence of microorganisms and procedure
on performing culture and gram staining. The proper waste management should be done and
precautions in the experiment was also included.
RESEARCH DESIGN
The pretest posttest experimental design was used in the study. A pretest posttest
design is an experiment where measurements are taken both before and after a treatment.

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Pretest-posttest designs are an expansion of the posttest only design with nonequivalent
groups, one of the simplest methods of testing the effectiveness of an intervention.
The design means that you are able to see the effects of some type of treatment on a
group. Pretest posttest designs may be quasi-experimental, which means that participants will
not be assigned randomly. Different Pretest-Posttest Designs are presented in a manner that
can help rehabilitation professionals to better understand and determine effects resulting from
selecting interventions (Allessandri, 2017). The experimental method of research was utilized
in the study for it is the best method of research to establish the cause and effect relationship
within the given hypothesis. The study was thrive to assess and observe the effect of Allium
sativum (Garlic) and Allium cepa (Onion) tubers ethanolic extract against Staphylococcus
aereus.
Sources of Data
The primary source of the data in the study are books, the related studies that were
gathered and if possible from available published journals. The Allium sativum (garlic) and
Allium cepa (onion) was used in the study that was conducted. This includes the tubers that
was extracted through ethanol.
Research Locale

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The Antimicrobial Effect of Allium sativum and Allium cepa was tested against
Staphylococcus aureus at DMMCIHS Medical Technology Laboratory. The said laboratory is
the place where most of the students are conducting their experiments.
Preparation of Culture Media
Suspend 28.0 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium
completely. Sterilize by autoclaving at 15lbs pressure (121ºC) for 15 minutes. If desired, the
medium can be enriched with 5-10% blood or other biological fluids. Cool to 45-50ºC. Mix
well and pour into sterile Petri plates.
Preparation of Materials Needed
The following materials that was used in the study is Beaker, Wooden Steering Rod,
Rubber Gloves, Damp Cloth, and Mortar Pestle, Cotton Swab, this is where you put all the
ingredients. Wooden Steering Rod, was used to mix the ingredients. Rubber Gloves was used
to protect the hands to any hazards while doing the experiment. Damp Cloth was used to
strain the extract of the garlic and onion. Mortar and pestle was used to crush allium sativum
(garlic) and allium cepa (onion) to get the extract. Cotton Swab was used to get a sample from
a surface.
Method and Procedure
Here are the phases in making Allium sativum (garlic) and Allium cepa (onion) Tubers
Ethanolic Extract to see its Antimicrobial effect against Staphylococcus aureus.

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In making the Allium sativum (garlic) and Allium cepa (onion) Tubers Ethanolic Extract
for its antimicrobial effect. First, is to collect some garlic and onion then dried it under the
sun, after drying it will be grinding using a blender and put it in a beaker, after that we put an
exact amount of ethanol in the beaker and wait until we get the extract of the garlic and onion.
Second, is to drained the garlic, onion and ethanol to get the extract, after that we measured
10grams of Nutrient Agar, in a beaker we put some water then add the 10 grams of Nutrient
Agar and mix thoroughly after it is mixed we put it in a four petri dish with the label of 100%,
75%, 50%and 25% and then we will air dry the Nutrient Agar and proceed to the next step.
Third, get four beaker with the label of 100%, 75%, 50% and 25%, in the first beaker which is
labeled as 100%, we add an amount of 100% of pure allium sativum and allium cepa extract
and the second beaker which is labeled as 75% we put an amount of 25% of distilled water
then add the 75% of pure allium sativum and allium cepa extract. While, on the third beaker
that is labeled as 50% we put 50% of distilled water and also 50% of pure allium sativum and
allium cepa extract, then the last beaker that labeled as 25% we put 75% of distilled water and
25% of pure allium sativum and allium cepa extract. Fourth, after mixing we inoculate all the
concentration to different surfaces such as tables, chairs and sink in the Medical Laboratory
Lab after air drying the surfaces wipe each of the surfaces using a cotton swab and put in the
petri dish with the agar. Lastly, it was incubated at 37 d C overnight. Growth of the organism
will be evaluated after 24 hours and will be recorded.
Preparation of Agar

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Nutrient Agar is a general purpose, nutrient medium used for the cultivation of
microbes supporting growth of a wide range of non-fastidious organisms. Nutrient agar is
popular because it can grow a variety of types of bacteria and fungi, and contains many
nutrients needed for the bacterial growth. Nutrient agar was used in culturing the organisms.
About 28 grams of NA was suspended in 1000 ml of distilled water. It was heated and boiled
to dissolve the medium completely. It was sterilized by autoclaving at 15 lbs pressure (121 d
C) for 15 minutes. The agar was then cooled at 40-50 d C.
Sterility test
Sterility test was performed by incubating the agar in incubator with 37 C temperature
for 18-24 hours and observed for no growth as indication for no bacterial growth.
Specimen Collection and Plant Extraction
Fresh Allium sativum and Allium cepa were collected in any barangay within Tanauan
City.The method was adapted to Olowa and Nuñeza (2013). The fresh plant was air dried for
about one week and ground into fine powder using a mechanical grinder. Twenty grams of the
fine powder of each plant samples were weighed and added into an Erlenmeyer flask
containing 250 mL of 95% ethanol. The solution was covered and shaken every 30 min for
about six (6) hours and allowed to stand for about 48 hours in room temperature. Then, it was
shaken and filtered using Whatman filter paper (No.1). After filtration, the solvent was
removed by evaporation using a rotary evaporator under reduced pressure at temperature
below 55°C. The extracts were reconstituted by dissolving in normal saline solution to the
required concentrations (100%, 75%, 50% and 25%). The reconstituted extracts were

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maintained at 2–8 ºC. The procedure was set to Medical Technology Laboratory at DMMC
Institute of Health Sciences for the testing and analysis.
Collection of Specimen and Culture
In conducting the study, samples collected were immersed in Normal Saline Solution and
were incubated. It wasstreaked at Nutrient Agar in different trials to check if there’s a
significant differences on the growth of organisms. It was incubated at 37 d C overnight.
Growth of the organism was then evaluated after 24 hours and was compared to positive
control and was interpreted.
Positive Control
The researchers used an antibiotic against S. aureus to determine the antimicrobial
effect of the Allium sativum (garlic) and Allium cepa (onion) Tubers Ethanolic Extract
Against Staphylococcus aureus.
Gram Staining Procedure
The organism isolated from the experiment was transferred into slide using inoculating
loop. It is then flooded using crystal violet staining reagent for about 1 minute. The slide was
washed into gentle by tap water for about 2 seconds and stained with a mordant (gram’s
iodine). The slide was washed again by tap water and decolorized for 15 seconds using
methanol. The slide was flooded with Safranin O as the counterstain for 30 seconds to 1
minute. Microorganisms was visualized under the microscope.
Proper Disposal and Handling of Specimen

P a g e | 26
Original cultures and sub-cultures and contaminated material are autoclaved at 121 º
Celsius for 15-30 minutes before disposal as regular waste material. Bacteria used in the study
were treated as if they were all pathogenic. Proper handling of specimen aree observed by
using safety equipment such as PPEs (Personal Protective Equipment). Cultures are
manipulated carefully to avoid uncontrolled release of aerosols or the generation of large
droplets or spills. Loading, removing, opening tubes and plates, streaking, sub-culturing was
done within a biological safety cabinet level 2.
Safety Precautions and Procedure
Bacteria are found everywhere in our environment, we are having contact with them on a
daily basis especially in the laboratory, while some of them are not pathogenic and have never
been shown to cause illness, under some circumstances a few of the microorganisms that are
not normally pathogenic can act as pathogens, therefore proper safety precautions will be
observed at all times. All bacteria was treated as potential pathogens. All materials like tubes,
plates, media, loops and other items were used for handling and culturing the bacteria was
sterilized by autoclaving. Operations and safe use of all materials and equipment were done.
Proper use of personal protective equipment like gloves, gown, mask, and goggles and
proper donning and doffing was observed at all times.10% bleach or 70% ethanol solution
was used to clean the work area before and after the handling of cultures. Eating or drinking is
strictly prohibited in the laboratory during handling of cultures. All cultures, chemicals and
media were labeled with their respective names, dates and hazard information. All items that
are discarded like tubes, culture plates, swabs, loops, and gloves are placed first in a

P a g e | 27
biohazard autoclave bag and autoclaved at 121 º Celsius for 15-30 minutes. Never eat or drink
in the laboratory while doing the experiment. Keep your fingers out of your mouth and wash
your hands before and after the laboratory experiment. Cover any cuts on your hands with a
bandage. Rubber gloves maybe worn as extra protection. The researchers segregated the
materials that were used in forming the allium sativum and allium cepa or garlic and onion
tubers ethanolic extract as an alternative medicine for its antimicrobial effect against
Staphylococcus aureus. First thing is sorting of reuse materials like beaker, wooden steering
rod, rubber gloves, and camphor powder. The researcher sterilized the beaker to get rid of
bacteria that remains in the beaker. Second, the rubber gloves and camphor powders that are
used is not that harmful to our environment, so it was disposed in canal. Lastly, the wooden
materials like wooden steering rod are sterilized and stored it in a storage room. An
unauthorized experiment or procedure must not do.
Statistical Tool
For SOP 1 the weighted mean was used for the level of the antimicrobial effect of
allium sativum (garlic) and allium cepa (onion) against staphylococcus aureus.
For SOP 2 T-test was used to determine if there any significant difference on the
level of antimicrobial effect of allium sativum (garlic) and allium cepa (onion) to antibiotic
(positive control) against S. aureus and as an alternative antibiotic in wound recovery.
Interpretation of Result

P a g e | 28
Growth was reported depending on the presence of microorganisms on the agar. Zero
(0) or no growth, 1 for slight growth, 2 for moderate growth and 3 for heavy growth of
microorganisms. By reporting significant levels will be determined. The growth on the culture
media will be differentiated by grading.

P a g e | 29
Plant
Identification and Authentication
Identification
Drying
Plant Cutting
collection Grinding
and Ethanolic Extraction
Extraction
Identification
Isolation
of Verification
Bacteria
Disposal
Waste Precautions
Handling
Interpretation
Statistical Analysis
of Result
Figure 2. Research Processs

P a g e | 30
Chapter III
Presentation, Analysis and Interpretation of Data
This chapter comprises the analysis, presentation and interpretation of the
results and findings from this study. The framework of the analysis and interpretation is
guided by the problems stated in Chapter 1.

Garlic
Amoxicillin
Concentration 100% 75% 50% 25% (Postive Control)
Trial 1
Procedure: 1
Gram Stain 3
(Prescence2 of cocci
2 were visualized
0 under the
Trial 2 1
microscope 4 on 3the microbial
based 4 1
effect of garlic and onion)
Trial 3
Microorganism Used: 0 3
Staphylocpccus aureus 4 3 1
Note: EffectiveOnion
in 100% concentration of both exctract.
Amoxicillin
Interpretation
Concentration 100% 75% 50% 25% (Postive Control)
0 No cocci seen
Trial 1 1 1 3 3 2
1 Slight presence of cocci
Trial 2 2 0 4 4 1
2 Slight to moderate presence of cocci
Trial 3 0 1 2 2 0
3 Moderate presence of cocci
4 Numerous cocci seen
(Garlic) and Allium cepa (Onion) tubers ethanolic extract against Staphylococcus aureus?
a. 100% b. 75% c. 50% d. 25%
Table 1. Statistical data results of (100%, 75%, 50% and 25%) concentration of allium
sativum (garlic) and allium cepa (onion) tubers ethanolic extract.

P a g e | 31
Paired Differences
95% Confidence
Interval of the
Std. Std. Sig.
Difference
Deviatio Error (2-
Mean n Mean Lower Upper t df tailed)
Pair garlic100 -.33333 .57735 . -1.76755 1.1008 - 2 .423
1 - 33333 8 1.000
onion100 4.000 2 .057
Pair garlic75 - 2.66667 1.15470 . -.20177 5.5351
2 onion75 66667 0 .000 2 1.000
Pair garlic50 - .00000 1.73205 1.000 -4.30265 4.3026 .000 2 1.000
3 onion50 00 5
Pair garlic25 - .00000 1.00000 . -2.48414 2.4841
4 onion25 57735 4
Table 1 shows that among the different concentration (100%, 75%, 50% and
25%) of garlic and onion, there is no significant difference between the antimicrobial
effect of allium sativum or garlic to allium cepa or onion in all concentrations.
(Garlic) and Allium cepa (Onion) tubers ethanolic extract against Staphylococcus aureus
to positive control?
Table 2.1 Statistical data result comparing 100%, 75%, 50% and 25% concentration of
garlic ethanolic extract to the positive control. Paired Samples Test

P a g e | 32
Paired Differences
Paired Differences 95%
95%
Confidence
Confidence
Interval of the Sig.
Std.
Interval of the
Difference (2-
Devi Std.
Std. Difference tail
atio Error Lowe
Mean Std. n Error Mean r Upp
Upper t df ed)
Mean Deviation Mean Lower er .000 2 1.00
Pair .00000 1.00 . - 2.4841
Pa 1 onion100 - .00000
garliccontrol 1.00000 .
000 57735 2.484 - 2.48 4 0
ir onioncontro
- 57735 2.4841 14 414
1 l garlic100 4
Pa Pair
onion75 - -.3333 1.15470 . . -- 2.53 - - 2 .015
- . 8.00
ir 2 onioncontro 3 66667 3.2017
2.6666 5773 33333 4.100 1.2324 510
garliccontrol 0
2 l 7 5 7
88 5
- garlic75
Pa onion50 - 2.0000 1.00000 . -.48414 4.48 - 2 .020
Pair garliccontrol - . . - -.89912
ir onioncontro 0 57735 414 7.00
3 - garlic50 2.3333 5773 33333 3.767
3 l 0
3 5 55
Pa onion25 - 2.0000 1.00000 . -.48414 4.48
Pair garliccontrol - . . - -.89912 - 2 .020
ir onioncontro 0 57735 414
4 - garlic25 2.3333 5773 33333 3.767 7.00
4 l
3 5 55 0
Table 2.1 shows that there is no significant difference between the antimicrobial effect of
garlic and the positive control in the 100% concentration. However, there is a significant
difference in the antimicrobial effect of garlic and positive control in 75%, 50% and 25%
concentration.
Table 2.2 Statistical data result comparing 100%, 75%, 50% and 25% concentration of
onion ethanolic extract to the positive control. Paired Samples Test
Sig.
(2-
d taile
t f d)
. 2 1.00
000 0
-.50 2 .667
0
DMMC INSTITUTE OF HEALTH AND SCIENCES 3.4 2 .074

#143 Narra St., Mt. view Subd., Brgy. 3, Tanauan City, Batangas 64
3.4 2 .074
64
P a g e | 33
Table 2.2 shows that among all the concentrations (100%, 75%, 50%, 25%) there is no
significant difference between the antimicrobial effect of onion and positive control of all
concentrations which means 100%, 75%, 50% and 25% are all effective.
3. Which concentration of Allium sativum (Garlic) and Allium cepa (Onion) tubers
ethanolic extract has the most antimicrobial effect against Staphylococcus aureus?
The research’s result shows that for the allium sativum or garlic tubers ethanolic
extract the concentration that has the most antimicrobial effect against Staphylococcus
aureus is the 100% concentration. While the other concentrations (75%, 50%, 25%)
concentrations have a significant difference to the positive control.
4. How may the findings of the study become helpful in the recovery of wound
infections?
The study will help the people especially those who live in the community
located in urban places to have an alternative medicine for wound recovery, the result
shows that the antimicrobial effect of garlic and onion is effective against the
Staphylococcus aureus which is a Gram-positive and round-shaped bacterium that is
commonly found in wound that often causes skin infections, pneumonia, heart valve
infections, and bone infections. Using garlic and onion as an alternative medicine can
help you save money which benefits those who cannot afford to buy such medications
and those far from the pharmacies. Aside from the antimicrobial effect of garlic and
onion there are also other benefits that these allium species could give for the wellness of
the human body.

P a g e | 34
Discussion
This study shows the Antimicrobial effect of allium sativum (garlic) and allium
cepa (onion) tubers ethanolic extract is effective and can be used against the Gram-
positive, round-shaped bacterium called staphylococcus aureus that can be isolated in
wounds. The experimentation took place at the Medical Technology Laboratory of
DMMCIHS. For the garlic among the different concentration (100%, 75%, 50% and
25%) compared to the positive control 100% has the most antimicrobial effect against
Staphylococcus aureus. For the onion compared to the positive control among the
different concentration (100%, 75%, 50% and 25%) all are effective and has the
antimicrobial effect against Staphylococcus aureus.
Chapter IV
SUMMARY, CONCLUSION AND RECOMMENDATION
This chapter present the result of the experiment. It also contains the
recommendation of the researchers on other ways or methods in doing this experiment as
well as their conclusions in based on the result of their experiment.
Summary

P a g e | 35
The researchers chose this study to experiment the antimicrobial effect of
allium sativum (garlic) and allium cepa (onion) tubers ethanolic extract against
Staphylococcus aureus as an alternative medicine to avoid wound infections. The
research’s result also showed that by using this onion and garlic against the
Staphylococcus aureus that can be found from wounds the medication is possible. It can
be also used in healing wounds faster. This is study can help people to save money and
use natural remedies like garlic and onion for wound.
The ethanolic extract from the allium sativum (garlic) and allium cepa (onion)
tubers can be an alternative medicine to avoid the wound infections. In the comparison of
the antimicrobial performance of onion to garlic, the result shows that there is no
significant difference in the antimicrobial effect of garlic and onion in different
concentrations.
Conclusion
This research concludes that the ethanolic extract taken from the allium species,
allium sativum or garlic and allium cepa or onion are effective and can be used against
staphylococcus aureus that causes wound infections. There is no difference between the
antimicrobial performance of garlic and onion in different concentration. For the garlic
(allium sativum) compared to the positive control the most effective is the 100 %
concentration and for the onion (allium cepa) compared to the positive control there is no
significant difference in all concentrations (25%, 50%, 75%, 100%) which means all
concentrations are mostly effective against S. aureus.

P a g e | 36
Recommendation
To the community, the researcher recommends to conduct activities that can show
the importance of onion and garlic especially to those urban places where anyone can
find many of this plant and make them familiar that onion and garlic extract can be
effective as an alternative medicine and can be used to avoid wound infections caused by
Staphylococcus aureus.
For the future researcher, this study could serve as a guide for them in conducting
their own research study and this data we gathered could also be used in developing their
own experimental study.

P a g e | 37
APPENDICES

P a g e | 38

P a g e | 39
NAME: Carol T. Galicia

STEM-12
ADDRESS: Pantay Matanda,
Tanauan City, Batangas
MOBILE NUMBER: 0946-002-4047
E-MAIL: [email protected]
________________________________________________________________
PERSONAL INFORMATION
Age: 17
Date of Birth: September 26,2003
Place of Birth: Mercado Hospital
Gender: Female
Height: 5'5
Civil Status: Single
Religion: Roman Catholic
Language Spoken: English, Filipino
Father's Name: Antonio P. Galicia
Mother's Name: Gina Silvina T. Galicia
________________________________________________________________
EDUCATIONAL BACKGROUND
SECONDARY: DMMC Institute Of Health science
2019-2020
Tanauan Institute
2016-2019
PRIMARY: Pantay Matanda, Tanauan City, Batangas

P a g e | 40
2011-2016
________________________________________________________________
NAME: Alexandra Almiñe
STEM-12
ADDRESS: Sn. Roque Sto. Tomas Batangas
________________________________________________________________
Age: 17
Date of Birth: July 29, 2003
Place of Birth: Sn. Roque Sto. Tomas Batangas
Gender: Female
Height: 5'2"
Father's Name: Eduardo A. Barit
Mother's Name: Alena A. Almiñe
________________________________________________________________
SECONDARY: DMMC Institute of Health science
2019-2020

P a g e | 41
Tanauan City Intergrated High School
2016-2019
PRIMARY: Tanauan North Central School
2011-2016
NAME: Cherryl V. Dimayuga
STEM-12
ADDRESS: Pantay Matanda, Tanauan City, Batangas
________________________________________________________________
Age: 17
Date of Birth: May 13, 2003
Place of Birth: Mercado Hospital
Gender: Female
Height: 5'2
Father's Name: Gerry O, Dimayuga

P a g e | 42
Mother's Name: Luningning T. Dimayuga
________________________________________________________________
2019-2020
Tanauan Institute
2016-2019
PRIMARY: Pantay Matanda, Tanauan City, Batangas
2011-2016
________________________________________________________________
NAME: Jignet Liann M. Reintegrado
STEM-12
ADDRESS: Poblacion Malvar Batangas
E-MAIL: jignetliannreintegrado@gmail/com
________________________________________________________________
Age: 18
Date of Birth: February 3, 2003
Place of Birth: Sto. Tomas general hospital
Gender: Female
Height: 5'3

P a g e | 43
Father's Name: Mark Lester E. Reintegrado
Mother's Name: Jennifer M. Reintegrado
________________________________________________________________
2019-2020
Malvar school of arts and trade
2016-2019
PRIMARY: Malvar Central School
2011-2016
________________________________________________________________
REFFERENCES
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Agarwal KC. Therapeutic actions of garlic constituents. Medicinal Research Reviews.
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Ankri S, Mirelman A (1999). Antimicrobial properties of allicin from garlic. Microbes
Infect., 2: 125-129. “Allium cepa,” IvyRose Holistic Health;
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"Allium sativum L". Kewscience; Plants of the World Online; Royal Botanic Gardens,
Kew, England. Retrieved May 26, 2017
Apata L (1979). Practice of Herbalism in Nigeria.University of Ife Press.
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Chemical, Agricultural and Medical Sciences (CAMS-2013) Dec. 29-30, 2013
Kuala Lumpur (Malaysia).
De Lucca, A. J., Bland, J. M., Boue, S., Vigo, C. B., Cleveland, T. E. and Walsh, T. J.
(2006a) Synergism of CAY-1 with amphotericin B and itraconazole.
Chemotherapy 52:285-287.
De Lucca, A. J., Boue, S., Palmgren, M. S., Maskos, K., and Cleveland, T. E. (2006b)
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relationship of structure and fungicidal activity. Canadian Journal Microbiology
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2013;27(3):380-383Department of Internal Medicine C, Rabin Medical Center,
Beilinson Hospital, Petah Tikva, Israel
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Biochimica et Biophysica Acta - General Subjects. 2013;1830:3670-3695
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against TB and other infections and economics of plant drugs—Introspection.
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Antimicrobial Effect of Allium sativum (Garlic) and

Allium cepa (Onion) Tubers Ethanolic Extract

Senior High Thesis Proposal Presented to the

In Partial Fulfillment of this Requirements

Jill Ivy A. Diamaro

The researchers’ beloved family

The researchers’ Capstone teacher Michelle Opulencia

The researchers’ statistician and class adviser Wennie Rose Matira

The researchers’ friends

The DMMC Institute of Health Sciences

The future researchers

And lastly, to the Almighty God.

DMMC INSTITUTE OF HEALTH AND SCIENCES

this thesis possible:

To Ms. Michelle Opulencia, the researchers’ Capstone teacher, for consistent

support and guidance in accomplishing this study.

the outcome of the experiment.

And most importantly, to the almighty God, for blessings, knowledge,

DMMC INSTITUTE OF HEALTH AND SCIENCES

THE PROBLEM AND ITS BACKGROUND

garlic in an experimental way.

understood. Further studies are needed to elucidate the pathophysiological mechanisms of

DMMC INSTITUTE OF HEALTH AND SCIENCES

the common red onion and is called Allium cepa.

family members, onions contain important health-promoting sulfur phytonutrients, such as

bacteria, fungi, viruses, and parasites. Alliums inhibit multi-drug-resistant microorganisms

and often work synergistically with common antimicrobials (Kyung, 2012).

DMMC INSTITUTE OF HEALTH AND SCIENCES

far from towns.

experiments and researches in college like medical technology.

Statement of the Problem

Ethanolic Extract against Staphylococcus aureus.

The following questions need to be answered:

1. Is there any significant difference on the antimicrobial effect of Allium sativum

aureus using the following concentrations?

DMMC INSTITUTE OF HEALTH AND SCIENCES

2. Is there any significant difference on the antimicrobial effect of Allium sativum

aureus to positive control?

ethanolic extract has the most antimicrobial effect against Staphylococcus

There is no significant difference on the antimicrobial effects of of Allium sativum

REVIEW OF RELATED LITERATURE

DMMC INSTITUTE OF HEALTH AND SCIENCES

. Nowadays, studies are being conducted by many researchers to discover new

have many potential scientific uses.

The benefits of garlic (Allium sativum).

It is scientifically proven that garlic is effectively used in cardiovascular diseases as a

patents on antimicrobial effect of garlic.

Other uses of Allium sativum.

DMMC INSTITUTE OF HEALTH AND SCIENCES

Resistance of Mycobacterium tuberculosis to antitubercular agents has called for an

Health discoveries of Allium sativum.

It is also of great importance because of its therapeutic properties and health-related

to explore its health-related potential (Hamidi, 2015). It is a historic medicinal plant,

recommended as dietary supplement (Martins, 2016).

DMMC INSTITUTE OF HEALTH AND SCIENCES

cardiovascular, neurodegeneration, cancer, and several other important diseases (Vasanthi,

including microbes (Bednareck, 2012).

Antimicrobial potential of Allium species.