Antibacterial Activity of

Garlic Extract
against Escherichia Coli
O157:H7

A Research Proposal

Presented to

Mrs. Lilia C. Tayco

at
Emiliano Tria Tirona Memorial National High
School

in
Partial Fulfillment of the Requirement in
Research I
 by

Nicole L. Cerenio
III-Mangga

November-December 2010

APPROVAL SHEET

In partial fulfillment of the course requirement in Research I,

this research paper entitled Antibacterial Activity of Garlic
Extract against Escherichia coli O517: H7 was prepared and
submitted to Mrs. Lilia C. Tayco, Emiliano Tria Tirona Memorial
National High School by Nicole L. Cerenio approved by the
committee on oral examination on with a grade of
________.

Panel of Examiners

Mrs. Lilia C. Tayco

Chairman

Mr. Celso L. Latosa

Mrs. Maria Paula G. Reyes

Member
Member
 Mr. Arnel P. Reyes

Chairman

Accepted in partial fulfillment of the course requirement in

Research I.

Recommended by:

Ms. Resurreccion Crisostomo Dr.

Presenciana E. Pinazo Ed. D.

Head Teacher VI, Science Department Principal

IV, ETTMNHS

ACKNOWLEDGMENT

The researcher would like to express his sincerest gratitude

and recognition to the following which made this research study
successful and worthwhile:

Foremost of all, to the ALMIGHTY GOD who gave him

strength and fortitude and served as his guiding light as he do
the whole research study;
 To his parents that served as his inspiration, for their moral
and financial support up to the very end and for their eternal
affection;

To Mrs. Lilia C. Tayco, his teacher in Research I, for her

whole-hearted support, conveyance and arousing motivations
that amplified his determination and confidence in finishing this
study; and

To his friends, classmates and fellow students who

never disappointed him in offering an overflowing support and
tremendous encouragement.

The researcher informs them his great appreciation for all

their help and participation because without having them, he
might not have accomplished this research study with all this
success and great satisfaction.

ABSTRACT
 The main objective of this research study is to get an
extract from garlic and test the activity of it to Escherichia coli
O157:H7 bacteria.

The antibacterial activity of a chemical constituent of garlic

was studied, identified and derived.

Experimental variables were tested using the portion of

organism constituent parts focus on cell-forming bacteria.

The results were tabulated, interpreted and verified after

the experimentation.

Microbiological cultures determine pathogenic organisms or

the type of organisms that may cause infectious diseases.
Incubation period of inoculated liquid broths with bacteria was
observed and analyzed.

One can determine the distinct advantage of frugality where

consumers are clamoring for cheap and common yet effective
kind of antibacterial product that can suit their health and
nutritional needs.
 DEDICATION

I, the researcher of this study, am conceitedly dedicating

this to:

To the ALMIGHTY GOD,

To my ever-supportive parents,

To my brothers and sisters,

To my relatives,

To Mrs. Lilia C. Tayco,

To my former science teachers,

To my friends, classmates and fellow students,

And to you who will be benefited in this study…

Thank you for all the heart-lifting motivations.

Expressing my gratitude,
TABLE OF CONTENTS
TITLE PAGE
APPROVAL SHEET
DEDICATION
ABSTRACT
TABLE OF CONTENTS
CHAPTERS:
I. THE PROBLEM AND ITS BACKGROUND
A. Introduction
B. Background of the Study
C. Statement of the Problem
D. Assumptions and Hypotheses
E. Significance of the Study
F. Scope and Delimitation
G. Definition of Terms
II. REVIEW OF THE RELATED LITERATURE
III. METHODOLOGY
 A. Materials
B. Equipment
C. Procedure
D. Flow Chart

BIBLIOGRAPHY

Chapter I

THE PROBLEM AND ITS

BACKGROUND

A. Introduction
Allicin is an organosulfur compound obtained from garlic, a
species in the family Alliaceae. Garlic has been used throughout
history for both culinary and medicinal purposes. The garlic
plant's bulb is the most commonly used part of the plant. With
the exception of the single clove types, the bulb is divided into
numerous fleshy sections called cloves.

Antibacterial activity of garlic extract was observed against

Escherichia coli which is known to be the cause of hemorrhagic
fever.
B. Background of the Study
Garlic is widely used around the world for its pungent flavor
as a seasoning or condiment. It is a fundamental component in
many or most dishes of various regions, including East Asia,
South Asia, Southeast Asia, the Middle East, Northern Africa,
Southern Europe, and parts of South and Central America. The
flavor varies in intensity and aroma with the different cooking
methods. It is often paired with onion, tomato, or ginger. It has
been proven that garlic has many uses.

Questions like “What might be the other uses of this plant

except for being a seasoning or a condiment” invades the
researcher’s intuitive mind.

C. Statement of the Problem

The scientific study aims to:

1. Get extract from fresh garlic.

2. Test the effect or activity of this garlic extract against the

growth of Escherichia coli.

D. Assumptions and Hypotheses

 Usually, when an ordinary person heard the word “garlic”,
the first thing that would come up into his mind is to sauté it with
meat or any related ingredient for it enhances the ingredient’s
flavor. The concept that the garlic has a content called allicin
came to the researcher’s mind after he had studied its
phytochemical contents and health benefits.

Hi: Garlic (Allium sativum) contains chemical constituent

called allicin.

Hi: Garlic (Allium sativum) extract really slowdown the

growth of Escherichia coli.

H0: Garlic (Allium sativum) extract has no distinct effect

against the growth of Escherichia coli.

D. Significance of the Study

The research study aims to determine the activity of garlic

extract against Escherichia coli. This study will be a great help to
those people who suffer from gastrointestinal diseases, in
particular, hemorrhagic fever. Most of the people nowadays are
less fortunate. Instead of buying expensive commercial
medicines, they may get extract from garlic which is available in
cheaper prices.

F. Scope and Delimitation

This research study mainly focuses on comparing the

growth of Escherichia coli on:

1. garlic extract and agar; and

2. controlled agar only.

The study and experimentation lasted from November to

December 2010 and was conducted at the researcher’s
residence, Block 15, Lot 13, Camia Street, Gardenville
Subdivision, Barangay Pag-asa 2, Imus, Cavite.

G. Definition of Terms
1. Garlic- a plant with a bulb shaped like an onion, which has a
strong taste and smell and is used in cooking

2. Allicin- is the key ingredient responsible for the broad-

spectrum of anti-bacterial activity in garlic

3. Extraction- is a separation process consisting in the

separation of a substance from a matrix

4. Extract- is a substance made by extracting a part of a raw

material, often by using a solvent such as ethanol or water

5. Growth medium or Culture medium- is a liquid or gel

designed to support the growth of microorganisms or cells, or
small plants like the moss Physcomitrella patens

6. Agar or agar-agar- is a gelatinous substance derived from a

polysaccharide that accumulates in the cell walls of agarophyte
red algae
 Chapter II

REVIEW OF THE RELATED

LITERATURE

Garlic (Allium sativum)

Allium sativum,
commonly known as garlic,
is a species in the onion
family Alliaceae. Its close
relatives include the onio n,
shallot, leek, chive, and
rakkyo. Garlic has been used
throughout history for both
culinary and medicinal
purposes. The garlic plant's
Garlic (Allium sativum)
bulb is the most commonly
used part of the plant. With the exception of the single clove
types, the bulb is divided into numerous fleshy sections called
cloves. The cloves are used for consumption (raw or cooked), or
for medicinal purposes, and have a characteristic pungent, spicy
flavor that mellows and sweetens considerably with cooking. The
leaves, and flowers (bulbils) on the head (spathe) are also edible,
and being milder in flavor than the bulbs, they are most often
consumed while immature and still tender. Additionally, the
immature flower stalks (scapes) of the hardneck and elephant
types are sometimes marketed for uses similar to asparagus in
stir-fries. The papery, protective layers of "skin" over various
parts of the plant are generally discarded during preparation for
most culinary uses, though in Korea immature whole heads are
sometimes prepared with the tender skins intact. The root cluster
attached to the basal plate of the bulb is the only part not
typically considered palatable in any form. The sticky juice within
the bulb cloves is used as an adhesive in mending glass and
china.

Allicin

Allicin is an organosulfur
compound obtained from garlic, a
species in the family Alliaceae. It was
first isolated and studied in the
laboratory by Chester J. Cavallito and
John Hays Bailey in 1944. This colorless
liquid has a distinctively pungent smell.
Chemical Structure of
Allicin
This compound exhibits antibacterial and anti-fungal properties.
Allicin is garlic's defense mechanism against attacks by pests.

Escherichia coli

Escherichia coli (commonly abbreviated E. coli;

pronounced /ˌɛʃɨˈrɪkiə ˈkoʊlaɪ/, named after Theodor Escherich) is
a Gram negative rod-shaped bacterium that is commonly found
in the lower intestine of warm-blooded organisms (endotherms).
Most E. coli strains are harmless, but some, such as serotype
O157:H7, can cause serious food poisoning and hemorrhagic
fever in humans, and are occasionally responsible for product
recalls. The harmless strains are part of the normal flora of the
gut, and can benefit their hosts by producing vitamin K2, and by
preventing the establishment of pathogenic bacteria within the
intestine.

E. coli are not always

confined to the intestine, and
their ability to survive for brief
periods outs ide the body makes
them an ideal indicator organism
to test environmental samples
for fecal contamination. The
bacteria can also be grown easily Escherichia coli
and its genetics are comparatively simple and easily manipulated
or duplicated through a process of metagenics, making it one of
the best-studied prokaryotic model organisms, and an important
species in biotechnology and microbiology.

E. coli was discovered by German pediatrician and

bacteriologist Theodor Escherich in 1885, and is now classified as
part of the Enterobacteriaceae family of gamma-proteobacteria.

Antibiotic Activity of Garlic

Louis Pasteur commented on the bactericidal (bacteria –

killing) effect of fresh garlic juice when dropped onto growing
bacterial colonies. Over the years studies have shown that fresh
garlic juice inhibits the growth of Staphylococcus (wound
infection), Brucella (brucellosis), Salmonella (Typhoid) and
several other bacteria. The action was comparable in vitro (in the
laboratory) with that of several antibiotics including penicillin,
streptomycin, chloramphenicol, tetracycline and erythromycin.

Agar

Agar or agar-agar is a
gelatinous substance derived from a

Putting an agar solution in

a petri dish
polysaccharide that accumulates in the cell walls of agarophyte
red algae. Historically and in a modern context, it is chiefly used
as an ingredient in desserts throughout Asia and also as a solid
substrate to contain culture medium for microbiological work.
The gelling agent is an unbranched polysaccharide obtained from
the cell walls of some species of red algae, primarily from the
genera Gelidium and Gracilaria, or seaweed (Sphaerococcus
euchema). Commercially it is derived primarily from Gelidium
amansii.

Agar (agar-agar) can be used as a laxative, a vegetarian

gelatin substitute, a thickener for soups, in jellies, ice cream and
other desserts, as a clarifying agent in brewing, and for paper
sizing fabrics.

Chapter III

Methodology

A. Materials

garlic cloves, agar powder, beef broth

B. Equipment
 mortar and pestle, cheese cloth, container, 10 petri dishes,
graduated cylinder, ruler, slide, microscope

C. Procedure

1. Preparation of garlic extract

a. Gather all the materials needed.

b. Pound the garlic cloves.

c. Cut the pound garlic into smaller pieces.

d. Put it into a cheese cloth.

e. Squeeze the cloth to get the garlic extract.

f. Store the extract into a clean and dry container.

2. Preparation of agar and Escherichia bacteria

culture

a. Boil water.

b. Add agar and beef broth.

c. Mix thoroughly.
 d. Transfer agar solution to 10 petri dishes.

e. Cool until gelatin consistency is attained.

3. Testing stage

a. Set aside 5 petri dishes for garlic extract environment

and 5 petri dishes for controlled environment.

b. Put 5ml of garlic extract on each 5 petri dishes.

c. Place a small sample of E. coli in the center of each

petri dishes.

d. Start timing the observation at daily intervals for 5

days.

e. Measure the diameter in centimeter (cm) of E. coli.

4. Microscopic Observation

a. Prepare slides of treated and untreated E. coli.

b. Examine under microscope.

C. Flow Chart
Extracting
Gathering
Measuring and
Storing
Agar Macroscopic
Microscopic
Incubation
Preparation Observ
the extract
Observation
Period
 BIBLIOGRAPHY

Internet Sources
 http://www.wikipedia.com
http://www.google.com
http://www.yahoo.com
http://www.encarta.com
http://www.eternalhealth.com
http://msn.com

Reference Books and Periodicals

Philippine Daily Inquirer
Magazines
New Book of Knowledge Encyclopedia
Webster Comprehensive Dictionary
Webster Comprehensive Thesaurus
People’s Journal