phytochemical screening of oregano against staphylococcus aureus.docx

5/22/2018 Phytochemical Screening of oregano against staphylococcus aureus.docx

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PHYTOCHEMICAL SCREENING OF OREGANO (Origanum Vulgare)AGAINST STAPHYLOCOCCUS AUREUS

In Partial fulfilment

Of the Requirements for the

Science Research II

Antipolo National High School

RAMOS, MARIFLOR

SOLOMON, MAILYN

TAMBAGO, JEAH MAE

TESARA, SHERNA

URBANO, CARISSA JOY

Proponents

February 2014


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Table of Contents

Title Page 1

Table of Contents 2

Chapter I

Background of the Study 4

Statement of the Problem 5

Hypothesis 5

Significance of the Study 6

Scope and Limitations 6

Definition of Terms 7

Chapter II

Review of Related Literature and Studies 8

Chapter III

Research Design 12

Setting of the Study 12

Subject of the Study 13

Procedure of the Study 14

Procedure of making Agar solution 15


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Procedure of the Experiment 15

Statistical Treatment 18

Chapter IV

Results and Discussion 20

Interpretation/Remarks 20

Chapter V

Findings 21

Conclusion 21

Recommendations 22


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CHAPTER I

BACKGROUND OF THE STUDY

Introduction

The researchers found some studies involving Phytochemical screening and its

benefits. Since then, it has been decided to conduct this screening to a very common

and versatile plant, oregano (Origanum vulgare). Aside from being an herbal medicine,

the researchers wanted to find out if it has properties which can be used to inhibit the

growth of Staphylococccus aureus. This bacterium is known to be a common cause of

skin infection. Because of this, the researchers extracted the leaves of oregano.

After the extraction, the two samples were tested on agar solution with Staphylococcus

aureus. The whole process of the research was conducted at Antipolo National High

School. Antipolo City.

Many plants can be found in the surroundings. They supply humans with food,

medicine, furniture and other materials. Some even have special characteristics that

can be utilized in various ways.

This project, Phytochemical Screening of Oregano (Origanum vulgare) Leaves

Extract against Staphylococcus aureusaims to utilize the use of oregano extract as an

inhibitor of Staphylococcus aureus. To be able to do this, the researchers opted to

undergo oregano to a Phytochemical screening to determine its properties and

medicinal benefits especially against the bacterium. Volatile oils such as thymol and

carvacrol can be found in the extract which is known to inhibit the growth of bacteria

such as Pseudonomas aeruginosaand Staphylococcus aureus.


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Oreganos (Origanum vulgare) extract have been used since the ancient times as

a disinfectant for wounds and to keep foods from different kinds of bacteria.

Conceptual Framework

The conceptual model is anchored on the relationship between the independent

and dependent variable of the study. The independent variable includes oregano

(Origanum Vulgare) extract in terms of its inhibition against Staphylococcus Aureus.

The other phase is the dependent variable which determines the properties that

oregano (Origanum Vulgare) extract may have that can inhibit Staphylococcus Aureus

and the rate of inhibition.

The line connecting the IV and DV determines if there is a significant relationship

between independent and dependent variables. The output of the study is

phytochemical screening of oregano(origanum vulgare)extract against staphylococcus

aureus.


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Figure 1

Conceptual Model on the Assessment of the phytochemical screening of oregano(Origanum Vulgare) extract against staphy lococcus aureus

INDEPENDENTVARIABLE

DEPENDENTVARIABLE OUTPUT

Oregano

(or iganum

vulgare) extract

phytochemical

screening

extract against

staphy lococcus

aureus

rate of inhibition

PHYTOCHEMICAL

SCREENING OF

OREGANO

(ORIGANUM

VULGARE)

EXTRACT AGAINST

staphy lococcus

aureus


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Statement of the Problem

The research aimed to determine the phytochemical contents of Oregano

(Oreganum vulgare) that can be used against Staphylococcus aureus. Specifically, it

seeks to answer the following questions:

1. What is the acceptability of Oregano (Oreganum vulgare) extract in

Phytochemical screening against Staphylococcus aureus?

2. In which rate of inhibition of Staphylococcus aureus exposed to 100% Oregano

(Oreganum vulgare) extract?

Hypothesis

This research aimed to test the null hypothesis that there is no significant

difference between the acceptability of Oregano (Origanun Vulgare) extract in

Phytochemical screening against Staphylococcus Aureus.

Significance of the Study

Medicine.Earth is abundant with plants that do not only provide for their food but

also shelter, furniture and clothing. Some plants even have special medicinal value just

like Oregano (Oreganum vulgare).

The study will be useful in inhibiting the bacteria, Staphylococcus aureuswhich

can be found in every human skin especially in the nostrils. It is responsible for the

formation of pustules or boils in the hair follicles. Being able to finish this study can do a

big help to eliminate the bacteria from our skin.


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Future Researchers. The result will serve as an input data for their future studies.

Scope and Limitations

The study aims to test the phytochemical components of Oregano (Origanum

vulgare) that can be used against the bacteria Staphylococcus aureus.

The study is limited to the Phytochemical screening of Oregano (Origanum

vulgare) to determine its contents responsible for inhibiting Staphylococcus aureus.

Furthermore, this was limited in testing the rate on inhibition of oregano extract

against Staphylococcus aureus.

Definition of Terms

For better understanding, the following terms were defined:

Oregano.A very abundant plant found in the Philippines. This has been the main

ingredient of the experiment where its extract was utilized.

Phytochemical. a naturally-occuring plant substance.

Screening. Test or testing carried out routinely. This involves the inhibition of the

staphylococcus aureususing oregano extract.

Staphylococc us aureus. often give rise to minor superficial diseases, including the

formation of pustules or boils in hair follicles; bacteria that was being tested upon with

the oregano extract.


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CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

This chapter presents various researches or studies in relation with Oregano

(Origanum vulgare) and Staphylococcus aureus that helps support this study.

Related Studies

The book entitled Beyond Antibiotics: Healthier Options for Families,by Michael

A. Schmidt, Lendon H. Smith and Keith W. Sehnert, has an entire section devoted to

the antimicrobial (antibacterial, antifungal and antiviral) efficacy of essential plant oils.

In this section they state that "one of the advantages essential oils have over

antibiotics is that bacteria do not develop resistance to essential oils." Many essential

oils exert their antibacterial effect by interfering with the bacteria's ability to breathe.

On the other hand, antibiotics interfere with the life cycle, or metabolism, of

bacteria, but since bacteria are very crafty creatures, they change their chemistry and

genes, which makes the antibiotic less effective the next time it is used. As a result, new

generations of antibiotics will need to be developed to stay ahead of these organisms.

Additionally, "another advantage to essential oils is that some actually stimulate immune

function."

Drs. E. Gildemeister and F.R. Hoffmann are referenced in the book as testing the

antimicrobial efficacy of certain essential oils as compared to a medium, phenol. Phenol

is an antiseptic substance found in Lysol, Pinesol and Chloraseptic throat spray. The

findings were quite a surprise to the investigators. Oil of oregano was the most potent


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antimicrobial essential oil tested by the investigators, as it proved to be 21 times

stronger than the medium.

Volume 18 of the British Naturopathic Journal included an article entitled "Kill and

Cure: The Healing Properties of Wild Oregano Oil." In the article, David Potterton, ND

summarized the findings of the world's foremost expert on oil of oregano, Dr. Cass

Ingram. According to Dr. Ingram, "oregano is the Rolls Royce of natural antiseptics". It

is, for example, far more active against noxious urinary pathogens than the typically

relied upon natural compounds such as garlic, goldenseal and echinacea.

The active ingredient, carvacrol, acts directly upon the mucous membranes of

the urinary tract and bladder. It offers the unique advantage of destroying both urinary

bacteria as well as yeasts, something that standard antibiotics fail to achieve." In

addition, according to the article, readers should get their hands on a copy of Dr.

Ingram's book The Cure is in the Cupboard: How to use Oregano for Better Health.

One of Dr. Ingram's predecessors, renowned scientist H. Martindale,

documented that the essential oil of oregano was the most powerful plant-derived

antiseptic known. This was back in 1910 when oregano was commonly used to treat

illness. Mr. Martindale demonstrated that oregano was 26 times more active as an

antiseptic than phenol, a powerful disinfectant used to sterilize hospital equipment.

In the article "A Serenade for Marinade" published July 5, 1999, in the Knoxville

News (Health and Science section), Dr. F. Ann Draughon, a University of Tennessee

microbiologist, studied the effects of herbs on food safety.

Dr. Draughon found that oil of oregano was the most effective at killing all


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pathogens tested against, including nine food-borne microbes: Listeria monocytogenes,

Staphylococcus aureus, Escherichia coli (commonly known as E-coli), Yersinia

enterocolitica, Pseudonomas aeruglinosa, Lactobacillus plantarum, Aspergillus niger,

Geotrichum and Rhodotorula.

Impressive results have been published about Oregano oil's qualities. Back in

1918, a French scientist named Cavel did research on meat stock cultured in septic

tank water. He found that Thyme oil and Oregano oil in very small doses were able to

destroy and prevent the development of microbes.

In 1992, two researchers at the University of Bologna, Italy (Caccioni and

Guizzardi) studied the fungicidal effects of essential oils on harvested fruits and

vegetables. They determined that carvacrol derived from Oregano oil was able to

almost completely inhibit spore development in fruits and vegetables at very minimal

doses.

Oil from the common herb oregano may be an effective treatment against

dangerous and sometimes drug-resistant bacteria, a Georgetown researcher has found.

Two studies have shown that oregano oil--and, in particular, carvacrol, one of oregano's

chemical components--appear to reduce infection as effectively as traditional antibiotics.

These findings were presented at the American College of Nutrition's annual meeting

October 6 and 7 in Orlando, Fla.

Harry G. Preuss, MD, MACN, CNS, professor of physiology and biophysics, and

his research team, tested oregano oil on staphylococcus bacteria--which is responsible


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for a variety of severe infections and is becoming increasingly resistant to many

antibiotics.

They combined oregano oil with the bacteria in a test tube, and compared

oregano oil's effects to those of standard antibiotics streptomycin, penicillin and

vacnomycin. The oregano oil at relatively low doses was found to inhibit the growth of

staphylococcus bacteria in the test tubes as effectively as the standard antibiotics did.

Another aspect of the study examined the efficacy of oregano oil and carvacrol,

which is believed to be the major antibacterial component of oregano, in 18 mice

infected with the staph bacteria

Six of the mice received oregano oil for 30 days, and 50% of this group survived

the 30-day treatment. Six received the carvacrol in olive oil, not oregano oil, and none

survived longer than 21 days.

Six mice received olive oil alone with no active agents (the control group) and all

died within three days. A repeat study corroborated these findings, which demonstrates

that there are components of oregano oil other than carvacrol that have antibiotic

properties.


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CHAPTER III

METHODOLOGY

Research Design

The study made use of experimental method of research in analyzing the

potential of Oregano leaves Extract in Phytochemical screening against

staphylococcus aureus.

Experimental design is directed by at least two hypotheses to determine

the expected relationship between two variables. The experiment was conducted to

confirm that the Oregano leaves extract is possible to use in Phytochemical

screening against Staphylococcus aureus.

The study is experimental since it is conducted to know if the extract of the

Oregano Leaves (Oreganum vulgare) is possible.

This method helps to determine the effectiveness of Oregano Leaves Extract

against Staphylococcus aurues and through this scientific method; other researchers

can use this study for the development of their researches.

Setting of the Study

The extraction process of the experiment was conducted in the residence of one

of the researcher at Northville Brgy. Bagong Nayon, Antipolo City. The testing was

performed in Solomons Residence using the Agar Solution at Brgy. Langhaya, Antipolo

City.


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Subject of the Study

The study used Oregano leaves (Oreganum vulgare) which are found in the

researchers backyard. It is called wild marjoram in many parts of Europe, it is small

shrub with multi branched stem covered with small greyish-green oval leaves. It grows

to 20 inches, with purple flowers and spade-shaped, olive-green leaves. It prefers a hot,

relatively dry climate, but will do well in other environments. To cultivate, it should be

planted in early spring, in fairly dry soil, with full sun. The plants should be spaced 12

inches apart.

Oregano Leaves (Oreganum vulgare) extract was used in Phytochemical

Screening against Staphylococcus aureus. Specifically, Oregano contains an

essential compound called Carvacol which proven effective to fight against Listeria

monocytogenesand Staphylococcus aureus.

Oregano is a perennial herb. The leaves, stems and flowers are used

medicinally. Oregano has been recognized for its aromatic properties since ancient

times. In ancient Greece, oregano was called 'Joy of the Mountain' and was

considered a symbol of joy and happiness. Ancient Egyptians

considered Origanumspecies to be sacred to the god Osiris, and wove it into

crowns or wreaths worn during rituals. Oregano is an important culinary herb, used

for the flavour of its leaves, which can be more flavourful when dried than fresh. It

has anaromatic, warm and slightlybitter taste, which can vary in intensity. Good

quality oregano may be strong enough almost to numb the tongue, but the cultivars

adapted to colder climates often have a lesser flavor. Factors such as climate,
http://pubs.acs.org/doi/abs/10.1021/jf0510079http://pubs.acs.org/doi/abs/10.1021/jf0510079http://pubs.acs.org/doi/abs/10.1021/jf0510079http://en.wikipedia.org/wiki/Aromahttp://en.wikipedia.org/wiki/Bitter_(taste)http://en.wikipedia.org/wiki/Bitter_(taste)http://en.wikipedia.org/wiki/Aromahttp://pubs.acs.org/doi/abs/10.1021/jf0510079http://pubs.acs.org/doi/abs/10.1021/jf0510079


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seasons and soil composition may affect the aromatic oils present, and this effect

may be greater than the differences between the various species of plants. Among

the chemical compounds contributing to the flavour

arecarvacrol,thymol,limonene,pinene,ocimene,andcaryophyllene.

Staphylococcus aureusis abacterium that is a member of theFirmicutes, and is

frequently found in the human respiratory tract and on the skin. Although S. aureusis

not alwayspathogenic, it is a common cause of skin infections, respiratory disease,

andfood poisoning. It is afacultative anaerobicGram-positivecoccalbacterium, also

known as "golden staph" and Oro staphira. Staphylococcus aureus belongs to the

family Staphylococcaceae. It affects all known mammalian species, including humans.

Further due to its ability to affect a wide range of species, S. aureus can be readily

transmitted from one species to another. This includes transmission between humans

and animals. S. aureusmay occur commonly in the environment.S. aureusis

transmitted through air droplets or aerosol. When an infected person coughs or

sneezes, he or she releases numerous small droplets of saliva that remain suspended

in air. These contain the bacteria and can infect others. Another common method of

transmission is through direct contact with objects that are contaminated by the bacteria

or by bites from infected persons or animals. Approximately 30% of healthy humans

carry S. aureusin their nose, back of the throat and on their skin.

Procedure of the Study

Prepare all the materials. Sterilize the materials. Collect oregano leaves. Then

wash it using clean water. Pound the leaves using mortar and pestle. Put on a clean
http://en.wikipedia.org/wiki/Carvacrolhttp://en.wikipedia.org/wiki/Thymolhttp://en.wikipedia.org/wiki/Limonenehttp://en.wikipedia.org/wiki/Pinenehttp://en.wikipedia.org/wiki/Ocimenehttp://en.wikipedia.org/wiki/Caryophyllenehttp://en.wikipedia.org/wiki/Bacteriumhttp://en.wikipedia.org/wiki/Firmicuteshttp://en.wikipedia.org/wiki/Pathogenhttp://en.wikipedia.org/wiki/Food_poisoninghttp://en.wikipedia.org/wiki/Facultative_anaerobic_organismhttp://en.wikipedia.org/wiki/Gram-positivehttp://en.wikipedia.org/wiki/Bacteriumhttp://en.wikipedia.org/wiki/Bacteriumhttp://en.wikipedia.org/wiki/Gram-positivehttp://en.wikipedia.org/wiki/Gram-positivehttp://en.wikipedia.org/wiki/Facultative_anaerobic_organismhttp://en.wikipedia.org/wiki/Food_poisoninghttp://en.wikipedia.org/wiki/Pathogenhttp://en.wikipedia.org/wiki/Firmicuteshttp://en.wikipedia.org/wiki/Bacteriumhttp://en.wikipedia.org/wiki/Caryophyllenehttp://en.wikipedia.org/wiki/Ocimenehttp://en.wikipedia.org/wiki/Pinenehttp://en.wikipedia.org/wiki/Limonenehttp://en.wikipedia.org/wiki/Thymolhttp://en.wikipedia.org/wiki/Carvacrol


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cotton cloth. Extract the pounded Oregano leaves in a beaker. Heat it for 3-4 minutes

using alcohol lamp, wire gauze and tripod. Transfer the extracted leaves in a 100-ml

graduated cylinder using funnel. Finally, shift it to the reagent bottle.

Procedure of making Agar Solution

Boil cup of water. Add 1 teaspoon of gelatine and 1 teaspoon of sugar. Stir for

1 minute until everything is dissolve. Then, cool the solution for 10 minutes. Fill a Petri

dish to a little below the top edge. Close the Petri dish by putting on the cover

immediately. Let the covered Petri dish stand for at least 4 hours.

Procedure of the Experiment

To determine the phytochemical screening of Oregano Leaves (Oreganum

vulgare) extract against Staphylococcus aureus. The researcher used the following

procedures:

A. Collection and Preparation of the Oregano Leaves


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Figure 2. Photographical Presentation of Oregano Leaves

B. Preparation and Strelization of the Material and Equipments

Figure 3. Photographical Presentation of the Materials and Equipments Used in

the study


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C. Washing the Oregano Leaves

Figure 4. Photographical Presentation of the Washed Oregano Leaves

D. Extraction of the Oregano Leaves

Figure 5. Photographical Presentation of Extraction Process in the Study


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E. Transferring the Product for Testing Purposes

Figure 6. Photographical Presentation of Transferring the Extract

Statistical Treatment

This formula is for computing the antimicrobial index

Al= Diameter of Clearing Zone - Diameter of Well

Diameter of Well


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CHAPTER IV

RESULTS AND DISCUSSION

Table 1. The Rate of zone of Inhibition of staphylococcus aureus upon exposure to

100% Oregano leaves extract

Test organism

Sample Clearing Zone, mm Antimicrobial

Index1 2

100% Oregano

Leaves Extract 18 15 0.88

Staphylococcus

aureus

Agar Solution

Interpretation/ remarks:

The extract from Oregano leaves boiled in 225 mL of water inhibited the growth

of staphylococcus aureus that was stocked at a room and observed after 24 hours.


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CHAPTER V

SUMMARY OF THE FINDINGS AND CONCLUSION

Findings

The findings of the study

1. * The rate of inhibition of Staphylococcus aureus upon exposure to 100%

Oregano Leaves Extract.

* The clearing zones on the well are 15 and 18, and the obtained antimicrobial

index was 0.88

2. The formulated assumption was proven because the potential of oregano leaves

extract ahs an antimicrobial potential that passed in Phytochemical screening against

Staphylococcus aureus.

Conclusion

Theres a significant difference between the acceptability of Oregano (Origanum

Vulgare) extract in Phytochemical screening against Staphylococcus aureus. And it is

concluded that Oregano Extract is possible to fight Staphylococcus Aureus.


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Recommendations

For more improvement of this study, the following are hereby recommended:

1. Make further experimentation in the potential of oregano

2. Conduct same study using the leaves of Oregano not by extracting

3. Further innovation on the Phytochemical Screening of Oregano Extracts are

encouraged

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