Columban College Barretto

Basic Education Department

Olongapo City 1

ANTIBACTERIAL PROPERTY OF Eleusine indica AND Rauvolfia serpentine

EXTRACT AGAINST Staphylococcus aureus

A Research Presented to the Faculty of Columban College-Barretto

Basic Education Department, Olongapo City

In partial Fulfilment of the Requirements for Capstone research.

by

LEIZL ROSE LABARES

MARIA MICHAELA C. MONTEVIRGEN
NIKKA MAE NILO

FEBRUARY 2019
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APPROVAL SHEET

In partial fulfillment of the requirements for the Capstone Research, this paper

entitled “ANTIBACTERIAL PROPERTY OF Eleusine indica AND Rauvolfia

serpentine EXTRACT AGAINST Staphylococcus aureus” has been prepared and

submitted by Leizl Rose Labares, Maria Michaela C. Montevirgen and Nikka Mae Nilo

is highly recommended for oral examination.

Mrs. Christine May Torres-Reyman

Adviser

Approved as partial fulfillment of the requirements for Practical Research on Oral

Examinations.

Panel’s name
Panel

Panel’s name Panel’s name

Panel Panel

Accepted in partial fulfillment of the requirements for the Practical Research

Ms. Christine May A. Torres - Reyman

Practical Research Adviser

FEBRUARY 2019
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ACKNOWLEDGEMENT

The researchers would like to express their sincerely gratitude to their adviser,

Mrs. Christine Mae Reyman for providing her invaluable guidance throughout the study

and Columban College Barretto for making this study possible.

Also we would like to thank to Ridon’s St.Jude Medical Center Laboratory for

helping in susceptibility test and Marica Pia D. Lico, Leah Fatima Cadavos and

Melody R. Lumasac of Department of Environment and Natural Resources ( DENR) for

identifying the plant material which have been used in the study.

And also other who supported the researcher and this study in any mean whether

moral or financial .
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ABSTRACT

Bacteria are organism that multiply quickly ,it can double within 20 to 30 minutes .

Some are good bacteria while most of them are pathogenic bacteria which can cause

diseases. And many are suffering from skin infection mostly because of Staphylococus

aureus. Plants has been used as an alternative medicine because of its capability to treat

such diseases particularly in skin. Many strains of Staphylococcus aureus bacteria have

become resistant to other traditional medicines. This study was carried out with an

objective to investigate the antibacterial potentials of leaves extract of Eleusine indica and

Rauwolfia serpentine. The aim of the study is to assess the antibacterial property and to

determine the possible zone of inhibition of extracts on Staphylococcus aureus strains and

to find an alternative way of utilizing the grass since they can be found anywhere so that it

won`t be place into waste. The extract was assessed using Steam Distillation process of

extraction. The antibacterial property was determined in the (,50, 100 μg/ml) extract

using agar disc diffusion method( Kirby-Bauer). The antibacterial activities of extracts

(,50, 100 μg/ml) of Eleusine indica and Rauwolfia serpentine were tested against

Staphylococcus aureus. Zone of inhibition of extract were compared with doxycycline,

linezolid and clyndamycine for antibacterial activity. Based on the test results obtained for

the sensitivity test of Eleusine indica and Rauwolfia serpentine extract, the concentration

of Eleusine indica and Rauvolfia serpentine leaves extract doesn’t exhibit inhibition effect

against Staphylococcus aureus .

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The researcher suggest that further study is required to prove that the crude extract and

the combination of the two plant which is Eleusine indica and Rauvolfia serpentine have an

antibacterial property against Staphylococcus aureus. The researcher also recommend to

use a research grade bacteria in performing antibacterial susceptibility testing.

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TABLE OF CONTENTS
Page
TITLE PAGE …………………………………………………….…...1

APPROVAL SHEET ………………………………………………….….…..2

ACKNOWLEDGEMENT ……………………………….……………….………..3

ABSTRACT …………………………………………………………4

TABLE OF CONTENTS …………………………………………………………6

CHAPTER

I. THE PROBLEM AND ITS BACKGROUND

Introduction . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

Statement of the Problem . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Significance of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

Scope and Delimitation of the Study . . . . . . . . . . . . . . . . . . . . . . . . . 13

II. THEORETICAL FRAMEWORK

Relevant Theories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Related Literature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

Related Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Conceptual Framework . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

Hypotheses of the Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

.
Definition of Variable . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
.
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IV. RESEARCH METHODOLOGY

Methods and Techniques of the Study . . . . . . . . . . . . . . . . . . . . . . . . . 24

Collection of Plant Materials… . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Preparation of Extracts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

Testing of Organism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Data Gathering Procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

V. PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA

Presentation, analysis and interpretation of data . . . . . . . . . . . . . . . . . 28

VI. SUMMARY OF FIDINGS, CONCLUSIONS AND RECOMMENDATIONS

Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Recommendations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

APPENDICES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33

BIBLIOGRAPHY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .34

CURRICULUM VITAE. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .36

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CHAPTER I

THE PROBLEM AND ITS BACKGROUND

Introduction

Bacteria can be found everywhere, they are present everywhere you stand, some are

beneficial but most of them are pathogenic in nature which means they are capable of

causing diseases. When species contains highly human pathogenic bacteria, it can cause

death. The use of medicinal herbs for curing disease had become popular until now not

only because of its more affordable compare to purchasing expensive pharmaceutical

medicines; it is also easy to gather. The increase in the population globally, possesses a

huge challenge to satisfy the needs of the people in terms of medicine. As the number of

affected people increases, the demand of medicine also increases. According to World

Health Organization (WHO) , the world is running out of supply of the antibiotic therefore

we need to keep researching/making new medicines that can cure to different kinds of

diseases. Large number of potential new drugs should suffice. There are several factors in

ASEAN countries which affect the availability of medicine in developing countries like

poor medicine supply and distribution systems; insufficient health facilities and staff; low

investment in health; and high cost of medicines.

In the Philippines, there are many people suffering from their diseases because they

cannot buy essentials medicine for their specific diseases, especially for those people who

are experiencing poverty and for those people who are unemployed.
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Plants are widely used in all over the world. Study of herbs has long been investigated

as antibacterial because it contains bioactive compounds which help in curing/healing

diseases. Because of their low toxicity, there is a long tradition of using dietary plants in

the treatment of infectious disease in Philippine folk medicine. Eleusine indica or Paragis,

also known by different names depending on the local dialects or places where it grows.

Eleusine indica or Paragis is an example of xerophytic weed which means it can adapt to

an environment which has small amount of water and usually it grows almost everywhere.

Local people from the Philippines are using paragis to cure certain health condition despite

of its lack of evidences and study. According to Department of Health of the Philippines

(DOH) paragis does not officially recognize it as a medicinal plant. Testing should be made

before it can be declared safe to use. The other herb which will be going to combined to

Eleusine indica is called Rauvolfia serpentine or commonly known as “snakeroot” it is

said that it is best remedy to cure such toxic bites. According to Dr. Gopi Krishna

Maddikera, the Eleusine indica is a plant growing rapidly and easily . And rural folks from

different places are also using it as an alternative medicine for certain ailments regardless

of its lack of studies because it can also be found almost everywhere.

This study is designed to assess in determining the antibacterial property of the

Eleusine indica and Ruvolfia serpentina extract against Staphylococcus aureus and to find

an alternative way of utilizing the grass since they can be found anywhere so that it won`t

be place into waste. We would also like to make a different and sample preparation of it

right in one`s home remedy aside from capsule and tablet forms. Lastly, we want to spread

the information of this study on how to supplement a nutritional need within in reach and

affordable in the community.

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Statement of the Problem

Generally, this study aims to assess the antibacterial property of Eleusine indica and

Rauvolfia serpentina extracts against Staphylococcus aureus. Specifically, it sought to

answer the following questions:

1. Is there any constituent present in the extracts of Eleusine indica and Rauvolfia

serpentine?

2.What is the zone of inhibition induced in Staphylococcus aureus using

2.1 Controlled negative

2.2 Controlled positive

2.3 Treatment A

2.4 Treatment B

2.5 Treatment C

3. Is there a significant difference among the controlled and treatment set-ups?

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Significance of the Study

The study of antibacterial property of Eleusine indica and Rauvolfia serpentine extracts

against Staphylococcus aureus will benefit the following:

Health Sector – the study will give an idea to health sectors on what way they will do in

case of emergency. If ever there is not enough medicine , they can use the techniques

mentioned in this research to give an immediate treatment to their patients. They can also

recommend the idea in this study to their patients especially to those people who live far in

the hospital. They can also use it as guide to implement a new kind of medicines using the

two plants.

 Patients- in this study the patients will benefit the most if the drug will be cheaper and

most especially it is safer. Through this research, they will be able to prevent more

expenses. Whenever, there is an urgent and the hospital is too far from them, they can use

this research to conduct first aid to prevent severe situation.

 Manufacturer- for further study in its quality in order to formulate and designed products

with an economical price. This research would benefit them because they would be able to

make use with the formulation of standards, guidelines and codes of ethical practice that

are possibly lead them to a new herbal medicine.

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 People living in the Rural Areas – the study will help the people living in rural areas for

them to support their needs of medicine without the use of money. It’s very applicable to

those people who cannot afford to buy expensive medicine.

 Educators and students- the study can provide and serve as useful topic for classroom

discussion and also serve as a guide to other related subjects regarding the study of plants.

It will stimulate the awareness of students on how they can use the combination of Eleusine

indica and Rauvolfia serpentine as the alternative medicine. It will help them to recognize

the importance of plants in our life. The techniques and methods mentioned in this research

will give idea to the students on the steps that they will do to make an alternative medicine

with the use of Eleusine indica and Rauvolfia serpentine. They can also use this research as

their basis if ever they will make a research like this.

 Future Researcher - This study is essential for future researches to make similar studies,

the ideas presented maybe use as a reference data that will give them a background study

and in conducting new researches or in testing the validity related to antibacterial activity

of Eleusine indica and Rauvolfia serpentine extracts. The researchers do believe that

through utilizing this paper as a source of data, scientific researcher will be encourage to do

more medicinal herbs which can help to each and everyone of us who are suffering from a

specific diseases. There might be an improvement with the promotion of healthy living for

preventing diseases as a result we can uplift the health care industry.

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Scope and Delimitation of the Study

This study will be limited only to the antibacterial property of Eleusine indica and

Rauwolfia serpentine’s stem and leaves extract. Other parts of the said plants will not be

use. The antibacterial analysis of Staphylococcus aureus were done at Ridon’s St. Jude

Medical Center. Concentrations other than those were not considered within the limits of

this study. Eleusine indica and Rauvolfia serpentine and its antibacterial property will be

the sole focus of this study. Other significant mechanisms or abilities of the said plant will

not anymore be contained within the limits of this research.

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CHAPTER II

THEORETICAL FRAMEWORK

This chapter deals with the identification of documents containing information

related to the study. Theories and studies gathered provide necessary information and

details that are significant and of relevance to the formulation of basic grounds of the

problems and possible solutions needed. In addition, it provides a deeper understanding

about the topic, thus making the proponent of the study more acquainted with the problem.

Analysis is done throughout the learning of the related studies and literature to which the

present study is correlated. Aside from that, this chapter also deals with the conceptual

framework which establishes the relationship between the variables and the definition of

variables used in the study.

Relevant Theories

Rauvolfia serpentine. In the article, “Integrative medicine: A clinician’s Journal”.

According to Douglas Lobay, Rauvolfia serpentine is a safe and effective treatment for

hypertension. It is also says that it was used by many physicians throughout India in 1940’s

and later use throughout the world in 1950’s including US and Canada.

It was used in folk medicine in India for about 2 and ½ decades to treat disease and

disorder such as abdominal pain, malaria and insect bites. And right after Vakel’s original

paper, more papers were distributed throughout the world ASEAN.

Rauvolfia serpentine’s medicinal properties are mainly attributed to the presence of

bioactive monoterpene indolealkaloids (MIAs) such as ajmaline , yohimbine , ajmalicine

,serpentine and reserpine. (Plummeret al., 1954; Bein, 1956;Lewis, 1956)

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Eleusine indica. Eleusine indica has many medicinal uses in the Philippines. It has been

used as antihelmintic and the decoction of the leaves used as diuretic in Surigao del Sur.

The whole plant that usually mixed with gogo are used to removed dandruff and to prevent

hair loss. . In Sablan, Benguet Province, decoction of leaves used for kidney problems and

arthritis while the decoction of its roots are used for fever. Ayta people from Porac,

Pampanga burn dried leaves and stems as repellent against hematophagous insects like

mosquito.

The different parts of the Eleusine indica were also used in different ways in other

countries. In Myanmar, Eleusine indica has used for hypertension. Aside from this,

Eleusine indica has been used as folklore medicine by our ancestors and even in other

Asian. Like in West Tropical Africa, the infusion of whole plant has been used for

hemopytosis. In Malaysia, the decoction of roots has been used for asthma. In Cambodia,

the plant has been used for fevers and liver complaints. In Nigeria, the leaves has been used

for treating diabetes and malaria. In Sri Lanka also, it has been used to treat sprained

muscle, wherein the roots or the entire plant was mixed with the scaped coconut and a

piece of Curcuma domestica is pounded and heated till cooked.

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Related Literatures

Eleusine Indica. In the International Journal of Drug Development and Research, it

was stated that ethanol extract of Eleusine indica has good effect to the early phase of

malaria. The Eleusine Indica is a medicinal shrub known in Africa and Asia. It is well

known because of the benefits it bring to the people specially those who experience

different kind of diseases. The Eleusine indica is known as paragis and sometimes

goosegrass. This annual grass belongs to family Poaceae usually grows in warmer areas. In

the Philippines, Eleusine indica usually used as herbal medicine . Leaves are used as

diuretic after boiling in Philippines(Gruyal et al. 2014) But sometimes the whole plant is

used to make an alternative medicine. This study was supported by another research

entitled Antimicrobial and Anthelmintic Activity that found the extract of Eleusine indica

can inhibit the intestinal worms

In the book entitled Therapeutic Use of Medicinal Plants and the Extracts,it has

been stated that decoction of plant of the goosegrass can be used in deworming( practiced

in Indonesia) ,coughs, lung troubles, bladder and kidney stones, spleen, liver complaints

and high blood pressure.

Rauvolfia Serpentina. Rauvolfia serpentine was commonly used in making a medicine in

India as stated in article of Roy( 1994). Its root is ground into powder and package in the

form of capsule. It is also reported that it can be used to treat high blood pressure. It was

proven in the result of Judson and Wilkin that it can lower blood sugar. It was also

mention in the article that serpentine is not only just for high blood, it can also be used in

treating hypertension.
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Rauvolfia serpentine has many medicinal uses because it can be used in such diseases

like snake bite,dysentery and regulatory problems. Because of presence of high amount of

alkaloids it is also used for the treatment of circulatory disorders ( Fabricant and

Farnsworth,2001), treat liver, abdomen pain, gastro internal disorders and to expel

intestinal worms from the children( nayak et al: Anisuzzaman et al.,2007; Dey and

Den,2011)

Rauvolfia in eastern medicine has many uses. It reduces dizziness and it also exerts

significant antagonistic effect on dopamine. Serpentine cause unpleasant side effects

commonly associated with antipsychotic medications, including problems with

coordination and stereotypic movements although it promote sedation and sleep. Serpentine

should only recommend occasionally to adolescent and not children because it can cause

adverse effects. In addition, serpentine is contraindicated during pregnancy, so if it is used

with adolescent females, they need to be counseled about the need to use contraception

(Medical Economics, 2007).

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Related Studies

Antibacterial Activity of combined medicinal plants extract against multiple drug

resistant strains. In the study entitled “Antibacterial Activity of combined medicinal

plants extract against multiple drug resistant strains”, the researchers asserts that that

combined extracts of Aegle Marmelos, Aphanamixis polystachia, Cuscuta reflexa and

Aesclynomene indica has antibacterial activity against Eschirichia coli, Klebsiella

pneumonia ,Proteus vulgaris and Pseudomonas aeruginosa. The study use disk diffusion

method to test the antibacterial propert of the said organisms.

Antibacterial Effect of Rauvolfia serpetina aqueous and ethanol extracts. In the

evaluation of the antibacterial effect of Rauvolfia serpentine aqueous and ethanol extracts,

the researchers found that ethanol extract exhibit greater antibacterial acitivity than the

aqueous extract against Staphylococcus aureus .It was shown from the evaluation of the

study that Rauvolfia serpentine has higher antibacterial effect when the extracts has a

component of ethanol than water.

Eleusine indica Possesses Antioxidant, Antibacterial and Cytotoxic Properties.

The Eleusine indica extract exhibited a broad spectrum antibacterial activity against all

tested bacteria except Bacillus subtilis, in which this bacterium was found to be resistant to

all Eleusine indica extracts. Meanwhile, hexane extract was demonstrated to have a

remarkable antibacterial activity against methicillin resistant Staphylococcus aureus

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(MRSA) and Pseudomonas aeruginosa, while the dicholoromethane extract did not exhibit

significant activity against P. aeruginosa.

Combined Effect of Crude Leaf Extracts of Selected Medicinal Plants against Selected

Enteric Bacterial Pathogens and Candida albicans. In the evaluation of the the effect

when the extracts of two plants combined, the researchers created different combinations

using 6 medicinal plants. Bulbine frutescens and Vernonia lasiopus produced less average

zones of inhibition when each of them is used separately against the microorganism .

However, the combinations of the plant extracts of; Aloe secundiflora and Bulbine

frutescens, Aloe secundiflora and Vernonia lasiopus, Bulbine frutescens and Vernonia

lasiopus, Bulbine frutescens and Tagetes minuta, showed increased antimicrobial activity

against Escherichia coli . This meant; the plant extracts from Bulbine frutescens and

Vernonia lasiopus when used in combination can enhance antimicrobial activity against

Escherichia coli. When the extracts of the plant were used in combinations they showed

diverse antimicrobial activity against the tested microorganisms. When the extracts of the

plants combined, there is a higher antibacterial activity against Eschericia coli compared

when each of those plants used separately against it. The result of this study described how

the combination of two extracts has a good effect to the antibacterial property of the plants

against human pathogen.

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Conceptual Framework

This study will use independent variable and dependent variable (IVDV) to show the

cause-effect relationship of two variables.

Independent Variable Dependent Variable

Eleusine indica and Antibacterial Activity of

Rauwolfia serpentine stem Staphylococccus aureus
extract

Figure 1. The research paradigm of the study

IV-DV model showing the effect of Eleusine indica and Rauwolfia serpentine extract to

Staphylococcus aureus

This study introduces combination of Rauvolfia serpentine and Eleusine indica

stem extract as the independent variable as shown in the first box. The second box

represents the antibacterial activity of Staphylococcus aureus as the dependent variable.

This framework illustrates the relation of two variables to each other and the observation of

what will happen during the study. This framework centers the inhibitory effect of Eleusine

indica and Rauwolfia serpentine extract against microorganism related to Staphylococcus

aureus through antibacterial assay.

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Hypothesis of the Study Tested

The researchers provided the hypothesis of this study, it is a testable prediction

concerning the relationship between the independent and dependent variables of this

study.

1. The extract of Rauwolfia serpentine has antibacterial property against

Staphylococcus aureus.

2. The extract of Rauwolfia serpentine doesn’t have antibacterial property against

Staphylococcus aureus.

3. The extract of Eleusine indica has antibacterial property against Staphylococcus

aureus.

4. The extract of Eleusine indica doesn’t have antibacterial property against

Staphylococcus aureus.

5. The combined extract of Eleusine indica and Rauwolfia serpentine has antibacterial

property against Staphylococcus aureus.

6. The combined extract of Eleusine indica and Rauwolfia serpentine doesn’t have

antibacterial property against Staphylococcus aureus.

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Definition of Variables

To provide a clearer understanding for this study, the following words are defined

operationally and conceptually.

Agar plates- are the standard solid support material for growing microorganisms.

Microbial growth media contains nutrients and an energy source to fuel the microbes as

they grow, and agar to keep the media in a semi-solid, gel-like state.

Antibacterial- anything that destroys bacteria or suppresses their growth or their ability to

reproduce. Heat, chemicals such as chlorine, and antibiotic drugs all have antibacterial

properties. Many antibacterial products for cleaning and handwashing are sold today. Such

products do not reduce the risk for symptoms of viral infectious diseases in otherwise

healthy persons.

Eleusine indica- a small annual grass distributed throughout the warmer areas of the

world. It is a tufted annual grass, prostrate and spreading, or erect to about 40 cm,

depending on density of vegetation but not usually rooting at the nodes.

Essential oils- are compounds extracted from plants. The oils capture the plant's scent and

flavor, also called its "essence." Unique aromatic compounds give each essential oil its

characteristic essence. Essential oils are obtained through distillation (via steam and/or water)

or mechanical methods, such as cold pressing.

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Extractions - are a way to separate a desired substance when it is mixed with others. The

mixture is brought into contact with a solvent in which the substance of interest is soluble,

but the other substances present are insoluble.

Kirby-Bauer – is an antibiotic testing (also called KB testing or disk diffusion antibiotic

sensitivity testing) uses antibiotic-containing wafers or disks to test whether particular

bacteria are susceptible to specific antibiotics. First, a pure culture of bacteria is isolated

from the patient.

Pathogenic- the types of germs that can cause some kind of disease. The flu, various

parasites, and athlete's foot fungus are all considered to be pathogenic. This word has been

used since the late 1800s to mean "producing disease," from the French pathogénique,

which in turn came from the Greek word for "disease," pathos.

Rauwolfia serpentine -also spelled ravolphia, is a medicinal shrub in the milkweed family.

Its root is ground into a powder and packaged in this form or sold in tablets or capsules. It

is a compound commonly used in Asian medicine, including traditional Ayurveda medicine

native to India.

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.

Steam distillation- is a separation process used to purify or isolate temperature sensitive

materials, like natural aromatic compounds. Steam or water is added to the distillation

apparatus, lowering the boiling points of the compounds. The goal is to heat and separate

the components at temperatures below their decomposition point.

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CHAPTER III

METHODS OF RESEARCH

This chapter presents the methods and techniques used in the study, the research

instrument, the data gathering procedures, data processing and statistical treatment utilized

in analyzing and interpreting the results of the study.

Methods and Techniques of the Study

This study use Quasi- Experimental method which fall under Quantitative type of

research as it focuses in numbers, logic and the gathering of date is structured. Quantitative

research have a statistical result that is good for estimating the probability of success.

According to Brian Craig Leonard, Quantitative research can be experimental

which subjects are measured before and after the treatment. The overall aim of

Quantitative research study is to classify features and construct statistical models to explain

what is observed.

Collection of Plant Material

Fresh Eleusine indica and Rauvolfia serpentine were collected at various areas from

Barretto, Olongapo City between January and February 2019 and were identified at

Department of Environment and Natural Resources (DENR) Upper Kalaklan, Olongapo

City. The specimen material was labeled, noted the date and the record of medicinal uses.
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Preparation of Extracts

The researcher used Steam Distillation Process of extraction to purify liquids that are

not very volatile and are immiscible with water.

During the steam distillation process, steam passes through the Eleusine indica and

Rauwolfia serpentine (separately) leaves and stem. The combination of heated steam and

gentle pressure causes the essential oil to be released and passes to a tube and pass to coil

connecting chamber. The essential oil rises to the top and is separated from the hydrosol

(floral water) and collected.

Figure 1
(Erich Handbook of Laboratory Distillation. New York: Elsevier Science Ltd., 1981)

Procedure:

1. Creating of own steam distillation apparatus set up

Materials:

•A heat source — usually direct fire

•A pressure cooker
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•A chamber with a hole that fits the copper tube

•A connecting copper tube to cool down and condense the steam flowing through the

pipe.

2. The plant material together with equal proportion of water is placed in pressure

cooker connected to a cold steam connecting chamber.

3. Bring the pressure cooker to a boil.

4. Keep an eye on the still to make you don’t run out of water and if the hot pipe

warms the water up. You need to replace it with another cold water or ice.

5. Once your distillation is complete, you pour the oil into a container for storage

as quickly as possible.

Note: Keep an eye on the whole process.

Figure 2 Figure 3
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Testing of Organism

The extracts of Eleusine indica and Rauvolfia serpentine was assessed against

Staphylococcus aureus which are acquired at clinical laboratory of Ridon’s St. Jude

Medical Center . Kirby-Bauer testing also known for disk diffusion assay was used in

determining the antibacterial property of Eleusine indica and Rauvofia serpentine extracts

to determine the sensitivity or resistance of bacteria. According to Jan Hudzicki (2009) ,the

presence or absence of growth around the disks was basis to measure the ability of extracts

to inhibit organism.

Data Gathering Procedure

Experimentation was the main tool used to gather data. It was conducted between the

two variables ,namely Rauvolfia Serpentina and Eleusine Indica , the independent variables

and Staphylococcus aureus, ,the dependent variable. The data was gathered through the use

of disk diffusion method conducted at Ridon’s St.Jude Medical Center wherein we’re able

to know the mean inhibition of the independent variable to dependent variable.

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CHAPTER IV

PRESENTATION, ANALYSIS AND INTERPRETATION OF DATA

This chapter presents the presentation, analysis and interpretation of the gathered

data on the study about the antibacterial property of Rauwolfia serpentine and Eleusine

indica through the help of Bacteriology section in the Ridon’s St. Jude Medical Center , the

combined extracts of Rauvolfia serpentine and Eleusine indica were tested in the

Staphylococcus aureus to determine the antibacterial property of Rauwolfia serpentine and

Eleusine indica. Disk diffusion method of susceptibility testing was used to analyse the

laboratory testing.

Figure 4. Result of Disk diffusion method of Susceptibility testing of the extracts

Rauwolfia Serpentina and Eleusine Indica against Staphylococcus aureus

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The figure 4 shows the result of the three extract which is the Eleusine indica extract,

Rauvolfia serpentine extract and the combined extract of Eleusine indica and Rauvolfia

serpentine that is tested to Staphylococcus aureus.The result show that the three extract

doesn’t exhibit inhibitory effect against the Staphylococcus aureus.

Results Obtained for the Sensitivity Test of Eleusine indica extract, Rauvolfia

serpentine extract and the combined extract of Eleusine indica and Rauvolfia

serpentine .

Bacteria: Staphylococcus aureus

Zone of

Control Extract Susceptible Intermediate Resistant Inhibition

Linezolid Rauvolfia x x ✓ No zone of

serpentine Inhibition

Cephalexin Eleusine x x ✓ No zone of

indica Inhibition

Doxycycline Rauvolfia x x ✓ No zone of

serpentine
And Inhibition
Eleusine
indica

The linezolid, cephalexin, and doxycycline antibiotic was used as a positive controlled in

the sensitivity test while the water was used as a negative controlled.
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Test Extract

A. The extract of Eleusine indica is resistant to Staphylococcus aureus. This means

that Eleusine indica leaves and stem etrcat has no zone of inhibition.

B. The extract of Rauvolfia serpentine is resistant to Staphylococcus aureus. This

means that Rauvofia serpentine stem and leaves extract has no zone of

inhibition

C. The extract of Eleusine indica and Rauvolfia serpentine is resistant to

Staphylococcus aureus. This means that Eleusine indica and Rauvolfia

serpentine has no zone of inhibition.

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CHAPTER V

CONCLUSIONS AND RECOMMENDATIONS

This chapter presents the conclusions and recommendations elicited on the

significant findings of the study.

Conclusions

From the findings, the following conclusions were drawn within the delimitations

of the study:

1.The concentration of Eleusine Indica and Rauvolfia Serpentina specie leaves

Extract doesn’t have an antibacterial activity against Staphylococcus aureus

2.The concentration of Eleusine Indica specie leaves extract doesn’t have an antibacterial

activity against Staphylococcus aureus

3.The concentration of Rauvolfia Serpentina specie leaves extract doesn’t have an

antibacterial activity against Staphylococcus aureus

4.The Linezolid, cephalixen and doxycycline antibiotic that is tested to Staphylococcus

aureus exhibit inhibitory effect . The combined extract of E. indica and R. Seroentina,

and also the negative control doesn’t showed any inhibitor effect against Staphylococcus

aureus.

5. There are significant difference between treatment and controlled set ups because the

positive control set up showed inhibition effect.

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Recommendations

For more reliable and accurate results, the research group should acquire pure

extract from Eleusine indica and Rauvolfia serpentine through

(1) the plant extract in varying concentrations submerged in water, methanol,

hexane or ethanol,

(2) perform phytochemical analysis on different plant extract concentrations,

(3) perform toxicity test

(4) grade bacteria is recommended in performing antimicrobial susceptibility

testing.
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Appendix 1

Documentation of Pictures

Plant Identification of Rauwolfia serpentina

Plant identification of Eleusine indica

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References

Al-Zubairi, A. S., Abdul, A. B., Abdelwahab, S. I., Peng, C. Y., Mohan, S., & Elhassan, M.

M. (2011). Eleucine indica possesses antioxidant, antibacterial and cytotoxic properties.

Evidence-Based Complementary and Alternative Medicine, 2011.

Archer, G. L. (1998). Staphylococcus aureus: a well-armed pathogen. Reviews of

Infectious Diseases, 26(5), 1179-1181.

Deshmukh, S. R., Ashrit, D. S., & Patil, B. A. (2012). Extraction and evaluation of insole

alkaloids from Rauwolfia serpentina for their antimicrobial and anti proliferative activities.

Int J Pharm Pharm Sci, 4(Suppl 5), 329-34.

Dey, A., & De, J. N. (2010). Rauvolfia serpentina (L). benth. ex Kurz.-A review. Asian

journal of plant sciences, 9(6), 285.

Elizabeth ,K .M.2001, Antimicrobial activity of Rauwolfia serpentine

Hayes, L. E. (1947). Survey of higher plants for presence of antibacterial substances.

Botanical Gazette, 108(3), 408-414.

Krell, Erich Handbook of Laboratory Distillation. New York: Elsevier Science Ltd., 1981.

Web site: http://www.essentialoil.com/products/glass-essencier-essential-oil-separator

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Lowy, F. D. (1998). Staphylococcus aureus infections. New England journal of medicine,

339(8), 520-532.

Monachino, J. (1954). Rauvolfia serpentina—Its history, botany and medical use.

Economic botany, 8(4), 349-365.

Negi, J. S., Bisht, V. K., Bhandari, A. K., Bisht, D. S., Singh, P., & Singh, N. (2014).

Quantification of reserpine content and antibacterial activity of Rauvolfia serpentina (L.)

Benth. ex Kurz. African Journal of Microbiology Research, 8(2), 162-166.

Rathi, P., Kumari, R., Rajput, C. S., & Sawhney, S. S. (2013). Therapeutic characteristics

of Rauvolfia serpentina. International Journal of Pharmacy and Pharmaceutical Sciences,

2(2), 1038-1042.

Srinava Murthy, K.N., Narayappa, M. 2015,In vitro study of antibacterial activity of leaf

and root extract of Rauvolfia serpentina against Gram Positive and Negative Bacterial

strain

Zaidan, M. R., Noor Rain, A., Badrul, A. R., Adlin, A., Norazah, A., & Zakiah, I. (2005).

In vitro screening of five local medicinal plants for antibacterial activity using disc

diffusion method. Trop biomed, 22(2), 165-170.

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CURRICULUM VITAE

PERSONAL DATA

Name : Maria Michaela C. Montevirgen

Address : Banaba St. Barretto Olongapo City

Date of Birth : October 13, 1999

Place of Birth : Taguig City

EDUCATIONAL ATTAINMENT

Elementary : Barretto II Elementary School

Secondary : Barretto National High school

AFFILIATIONS (Membership and Officership to Organization/s)

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PERSONAL DATA

Name : Nikka Mae Nilo

Address : Abra St. Barretto Olongapo City

Date of Birth : October 7, 2000

Place of Birth : Olongapo City

EDUCATIONAL ATTAINMENT

Elementary : Bacong Elementary School

Secondary : Barretto National High School

AFFILIATIONS (Membership and Officership to Organization/s)

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PERSONAL DATA

Name : Leizl Rose Labares

Address : Rizal ext. Barretto Olongapo City

Date of Birth : July 7,2001

Place of Birth : Iligan City

EDUCATIONAL ATTAINMENT

Elementary : Tambo Central Elementary School

Secondary : Iligan City National High School

AFFILIATIONS (Membership and Officership to Organization/s)