Antibacterial Activity of Garlic Extract Against E. Coli

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 ________.

Panel of Examiners

Mrs. Lilia C. Tayco Chairman

Mr. Celso L. Latosa Mrs. Maria Paula G. Reyes Member Member

with a grade of

Mr. Arnel P. Reyes Chairman

Accepted in partial fulfillment of the course requirement in Research I.

Recommended by:

Ms. Resurreccion Crisostomo Presenciana E. Pinazo Ed. D.

Dr.

Head Teacher VI, Science Department IV, ETTMNHS

Principal

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

residence,

Block

15,

Lot

13,

Camia

at

Street,

Subdivision, Barangay Pag-asa 2, Imus, Cavite.

G. Definition of Terms

the

researcher’s Gardenville

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 broadspectrum 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 bulb is the most commonly

Garlic (Allium sativum)

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 compound

is

an

obtained

organosulfur 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 Macroscopic Observ Storing Agar Microscopic Incubation Preparation

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