Muntinlupa Science High School

999 Buendia St., Tunasan, Muntinlupa City

The Efficiency of Paragis (Eleusine indica) as an Antibacterial

Extract Against Staphylococcus aureus

A Science Investigatory Project

LIFE SCIENCE
(Team)

Presented
to
Department of Education (DepEd)
Muntinlupa City

Project Proponents
NICOLE JESSLY AGULLANA
DANIELLE ESCOLANO
KYLA ORBINA
KATELYN VALERA

Research Adviser
Ms. Jenica D. Lopez

Research Adviser
Ms. Webena F. Gereno
Mr. Ryan Jumar Pantoja

1
 TABLE OF CONTENTS

Page

Title Page …………………………………………………………… 1

Research Plan …………………………………………………...... 3

Acknowledgement ………………………………………………… 7

Abstract …………………………………………………………..... 9

Chapter I: INTRODUCTION …………………………………….... 10

Background of the Study

Statement of the Problem

Objectives

Hypothesis

Significance of the Study

Scope and Delimitations of the Study

Definition of Terms

Conceptual Framework

Chapter II: REVIEW OF RELATED LITERATURE ……….......... 15

Chapter III: METHODOLOGY ………………………………......... 19

Chapter IV: RESULTS AND DISCUSSION …………………....... 22

Chapter V: SUMMARY, CONCLUSION, AND RECOMMENDATIONS 24

Bibliography …………………………………………………………. 26

ISEF Official Abstract ……………………………………………….

Intel ISEF Forms …………………………………………………….

Plagiarism Check Report …………………………………………..

Scanned Logbook …………………………………………………..

2
 Research Plan

Rationale

Paragis or Eleusine indica is a weed that is abundant in many places

that can withstand temperatures as low as -7 degrees. It is native to Asia,

Africa, and Papuasia. Though it has not been scientifically studied and

verified, many people in rural areas use Paragis or E. indica to treat arthritis,

diabetes, wounds, fever, hypertension, and many more because of its many

medicinal properties.

Although the weed Paragis or E. indica is not yet claimed or studied

scientifically, many people, especially those from rural areas, are claiming that

Paragis has antibacterial properties that helps in wound treatment. With this,

our study’s aim is to utilize the antibacterial properties of Paragis or E. indica

extract, specifically against Staphylococcus aureus, which is a gram-positive

bacteria that spreads through an infected person. This way, we would be able

to open new alternatives for the mass, especially in terms of wound healing,

and prevention of infections from opportunistic bacteria in open wounds

through proper experimentation by antimicrobial sensitivity tests.

Statement of the Problem

The main target of this research is to utilize the antibacterial properties

of Paragis or E. indica by means of experimentation and cross-analysis of its

advantages and disadvantages as an antibacterial extract.

3
Main Questions:

1. Was the utilization of Paragis or E. indica effective as an antibacterial

extract?

2. What was the effect of the Paragis or E. indica extract on the S. aureus?

Significance of the study

This study will be able to open an alternative way for people to utilize

the properties of the plants around them, specifically the antibacterial

properties of Paragis or E. indica, which can be found anywhere. This

research is relevant and timely as it not only benefits them but the whole

population in general. This is because it addresses infection, which is

something that we all have faced at a certain point, regardless of where it

comes from (e.g. open wounds). With the implementation of this study, the

different properties of Paragis or E. indica will also be open to utilization in

many fields.

Objectives

At the end of the study, the researchers would be able to:

1. Utilize the antibacterial properties of Paragis or E. indica.

2. Find evidence that Paragis or E. indica can be an efficient and

effective antibacterial extract.

Hypothesis

Null Hypothesis:

4
 The Ethanol extract of Paragis or E. indica as an antibacterial extract

has no significant effect on S. aureus that can be found on minor cuts and

wounds.

Alternative Hypothesis:

The Ethanol extract of Paragis or E. indica as an antibacterial extract

has a significant effect on S. aureus that can be found on minor cuts and

wounds.

Methodology

The researchers would first have Ethanol Soxhlet Extraction performed

on Paragis or E. indica to get the Paragis extracts, which should be in

different concentrations. The S. aureus will be provided by a university

laboratory, along with the other needed equipment. The researchers will then

prepare the materials needed for the experimental set-up which include 3

Petri dishes, 100 grams of Lysogeny Broth Agar, 3 packs of 25 grams of

Gelatin Powder, 5 liters of Distilled Water and 100 grams of Dextrose Powder,

S. aureus, paper discs, and the different solutions of the Paragis extract. T he

researchers will proceed in creating the LB Agar by mixing of 9g of Gelatin

powder, 2.5g of LB Broth powder and 250 ml of boiled distilled water in a

sterile flask. After that, the researchers will equally distribute the broths into

the 3 Petri Dishes, the excess agar is left in an Erlenmeyer flask or container,

and wait for the agar to cool and harden. After the broths solidified and turned

into agar, the researchers will then prepare the following materials which

include an alcohol lamp, inoculation loops, gloves, and other equipment

needed when inoculating. Then, the researchers will use a syringe to suck the

5
S. aureus and apply the bacteria to the Petri dishes then streak the plate in

order to spread the S. aureus. After 24 hours of incubation at 37 degrees

Celsius, the researcher will proceed to get the results of the experiment by

measuring mainly the zone of inhibition as basis for the bacteria’s

susceptibility or resistance.

Risk and Safety

Our study involves hazardous component that might cause harm and

danger just as the bacteria used in the series of tests. According to the MSD

Manual, Staphylococcus aureus is the most dangerous of all of the many

common staphylococcal bacteria. These gram-positive, sphere-shaped

(coccal) bacteria often cause skin infections but can cause pneumonia, heart

valve infections, and bone infections.

6
 ACKNOWLEDGEMENT

We would like to express our deepest appreciation to the people who

helped us bring this study to life. With this, we would like to thank and extend

our gratitude to the following:

To Grade 10 – Ptolemy, for their support throughout the whole

research process. We would have never pushed through without them.

To Sir Jan Jeremiah Unas, our Grade 8 Biotechnology Teacher, for

helping us with the whole experimentation process, making sure that we were

guided through each step of the way.

To Sir Ryan Jumar Pantoja, our Research Consultant and Class

Adviser, whose never-ending support and guidance helped us bring this study

to life.

To Ma’am Webena Gerona, our school’s Science Coordinator, whose

constructive criticism, and suggestions helped us better our study.

To the Adamson University Technology Research and Development

Foundation, for helping us with the Ethanol Soxhlet Extraction.

To Laguna State Polytechnic University – Los Banos’s Sir Christian

Dela Cruz, Sir Glenn Adrian Paypon, Ms. Precious Tolentino, Sir Rupert

Olivares, and Sir Rainiel Domasion, for guiding us throughout the whole

experimentation process, and helping us better our study.

To Ma’am Jenica Lopez, our beloved Research adviser, whose never-

ending guidance and patience motivated us to pursue and finish this research

paper.

7
 To Dr. Emelita D. Bautista, our school principal, for her whole-hearted

support in the accomplishment of this study.

To our parents, for always providing us everything that we needed to

complete this research study, and at the same time, for encouraging us to do

our best.

Above all, to our Almighty God, for being with us through thick and

through thin, to whom we pass down our worries and frights. Thank you for

this academic endeavor, where we were able to learn so much. To God be all

the Glory.

8
 ABSTRACT

Paragis is a plant that is commonly found in the Philippines. Since

Paragis is a weed, it is usually ignored by the Filipino people especially the

ones who are living in the city. According to our research, Paragis is usually

used as a remedy in the rural parts of the country but it isn't scientifically

proven whether it has medicinal properties or not.

The researchers came up with the idea to try and find if it does have

antibacterial properties. The researchers found out that there had been

studies that proved that Paragis has components in it that is used in the

medical field.

The Paragis is really easy to find, we can find it almost anywhere in our

community. The Paragis went through a process called Soxhlet Ethanol

Extraction. With regards to the experimentation process, which is the Kirby-

Bauer Susceptibility Test, we found out that the Paragis extract was

ineffective against Staphylococcus aureus.

9
 CHAPTER I

INTRODUCTION

Background of the Study

Paragis or Eleucine indica is a weed that is abundant in in many places

that can withstand temperatures as low as -7 degrees. It is native to Asia,

Africa, and Papausia. Though It has not been scientifically studied and

verified, many people in rural areas use Paragis or E. indica to treat arthritis,

diabetes, wounds, fever, hypertension, and many more because of its many

medicinal properties.

Although the weed Paragis or E. indica is not yet claimed or studied

scientifically, many people, especially those from rural areas, are claiming that

Paragis has antibacterial properties that helps in wound treatment. With this,

our study’s aim is to utilize the antibacterial properties of Paragis or E. indica

extract, specifically against Staphylococcus aureus, which is a gram-positive

bacteria that spreads through an infected person. This way, we would be able

to open new alternatives for the mass, especially in terms of wound healing,

and prevention of infections from opportunistic bacteria in open wounds

through proper experimentation by antimicrobial sensitivity tests.

Statement of the Problem

The main target of this research is to utilize the antibacterial properties

of Paragis or E. indica by means of experimentation and cross-analysis of its

advantages and disadvantages as an antibacterial extract.

Main Questions:

10
1. Was the utilization of Paragis or E. indica effective as an antibacterial

extract?

2. What was the effect of the Paragis or E. indica extract on the S. aureus.

Objectives

At the end of the study, the researchers would be able to:

1. Utilize the antibacterial properties of Paragis or E. indica.

2. Find evidence that Paragis or E. indica can be an efficient and

effective antibacterial extract.

Hypothesis

Null Hypothesis:

Paragis or E. indica as an antibacterial extract has no significant effect on S.

aureus that can be found on minor cuts and wounds.

Alternative Hypothesis:

Paragis or E. indica as an antibacterial extract has a significant effect on S.

aureus that can be found on minor cuts and wounds.

Significance of the Study

This study will be able to open an alternative way for people to utilize

the properties of the plants around them, specifically the antibacterial

properties of Paragis or E. indica, which can be found anywhere. This

research is relevant and timely as it not only benefits them but the whole

11
population in general. This is because it addresses infection, which is

something that we all have faced at a certain point, regardless of where it

comes from (e.g. open wounds). With the implementation of this study, the

different properties of Paragis or E. indica will also be open to utilization in

many fields.

Scope and Delimitations of the Study

This study will only focus on the efficiency of the Paragis (E. indica)

extract against S. aureus through its zone of inhibition.

However, due to lack of sufficient time and resources, the experiment

will only be determined using qualitative/descriptive analysis and not through

quantitative means (Bacterial CFU). Also, there is a limited concentration with

regards to the Paragis (E. indica) extract, so its efficiency will be measured

with only one concentration.

Definition of Terms

Paragis (Eleucine indica) extract - it is an extract which is believed to have

anti-bacterial properties that will be used in the study against Staphylococcus

aureus.

Staphylococcus aureus - it is a gram-positive, sphere-shaped bacteria that

spreads from person to person by direct contact; also known as an

opportunistic bacteria.

Lysogeny Broth Agar - it is the most widely used nutritionally rich medium

primarily used for the growth of bacteria.

12
Bacterial CFU - also known as colony forming units, refers to individual

colonies of bacteria, yeast, or mold. It is a mass of individual cells of the same

organism, growing together.

Ethanol Soxhlet Extraction - The Soxhlet extraction process heats the

solvent (ethanol) to boiling temperature (>78°C). The evaporated ethanol is

contained within the apparatus by the condenser unit; however the apparatus

should be placed under a fume hood in case of escape.

13
Conceptual Framework

Gathering

Paragis Staphylococcus
aureus

Ethanol Soxhlet
Culturing
Extraction

Experimentation
(Kirby-Bauer Antibiotic Susceptibility Test)

1st Trial 2nd Trial 3rd Trial

Positive Control Positive Control Positive Control

(Streptomycin) (Streptomycin) (Streptomycin)

Negative Control Negative Control Negative Control

(Distilled Water) (Distilled Water) (Distilled Water)

Experimental Experimental Experimental

Control Control Control
(Paragis) (Paragis) (Paragis)

Collection of
Results

14
 CHAPTER II

REVIEW OF RELATED LITERATURE

Review of Related Literature

According to Ecophysiological Studies of Eleusine indica (L.) Gaertn.

and Sporobolus pyramidalis P. Beauv. at Ibadan, NigeriaE. “Eleusine indica,

also known as fowl-foot grass, has been described by Purseglove (1975) as a

well-known species throughout Africa and Asia. It is now common in the

tropics of both hemispheres. After careful observations, it has been found out

that the population of E. indica is affected by grazing in pre-flowering

condition. The spread of the E. indica depends mainly on seeds. McIllroy

(1964) said that E. indica thrives well in trampled areas and becomes really

common along areas where cattles had travelled.”According to Eleucine

indica Possesses Antioxidant, Antibacterial and Cytotoxic Property, “E.indica

is considered to be a wild plant because of its availability everywhere, it can

also be found on tropical and subtropical regions. Paragis (E. indica) could

survive different kinds of its environment, but its growth is reduced during dry

seasons. The whole plant, especially its root, can remove contaminants,

increase urine production, reduce fever and increase bowel movement to

prevent constipation, and that’s why it is used for the treatment of influenza,

hypertension, urine output and retention. The seed is sometimes used as food

when famine strikes and is also used in treating problems in the liver.”

Review of Related Legal Basis

15
 Republic Act No. Republic Act No. 3720, knowns as "Food, Drug, and

Cosmetic Act" is an act to ensure the safety and purity of foods, drugs, and

cosmetics being made available to the public by creating the food and drug

administration which shall administer and enforce the laws pertaining thereto.

It is hereby declared the policy of the State to insure safe and good quality

supply of food, drug and cosmetic, and to regulate the production, sale, and

traffic of the same to protect the health of the people.

Republic Act No. Republic Act No. 9502, knowns as “Universally

Accessible Cheaper and Quality Medicines Act of 2008” is an an act providing

for cheaper and quality medicines, amending for the purpose republic act no.

8293 or the intellectual property code, republic act no. 6675 or the generics

act of 1988, and republic act no. 5921 or the pharmacy law, and for other

purposes.

Republic Act No. Republic Act No. 7394, knowns as “Consumer Act

of the Philippines” is the policy of the State to protect the interests of the

consumer, promote his general welfare and to establish standards of conduct

for business and industry.

Republic Act No. 2067, known as the "Science Act of 1958", In

consonance with the provisions of section four, Article XIV of the Constitution,

it is hereby declared to be the policy of the state to promote scientific and

technological research and development, foster invention, and utilize scientific

knowledge as an effective instrument for the promotion of national progress.

Through the defined use of these laws, the main aim of the research

is to promote scientific and technological research and development, foster

16
invention, and utilize scientific knowledge to students, and to promote organic

agriculture and propose an accessible cheaper and quality medicines as

means of creating a fruitful product or innovation.

when famine strikes and is also used in treating problems in the liver.”

Review of Related Studies

Local

Throughout the different parts of the world, especially in Asia, Paragis

is used by many people, especially those in rural communities, to cure

diseases or ailments. In a review article entitled “Paragis : What can you get

from it?” (Eslit, 2018), Paragis is said to have curative powers, and it contains

many important properties, with it being anti-inflammatory, antibacterial,

antioxidant, cytotoxic, etc. Paragis is also said to to offer a lot of health

benefits to mankind, but the country’s Department of Health (DOH) is hesitant

to declare it as a recognized medicinal plant as exhaustive research and

testing should be done first.

Foreign

A study conducted by Odomena, Effiong, and Udobang from the

International Journal of Drug Development and Research entitled

“Antiplasmodial and Antidiabetic Activities of Eleusine Indica,” was also able

to give a review about the phytochemicals or Paragis, and how its many

properties, with an emphasis on it being antibacterial, antiplasmodial, and

antidiabetic, helps in curing many ailments along the different parts of the

world.

17
 Another study conducted by Iqbal and Gnanaraj from the

Environmental Health and Preventive Medicine entitled “Eleusine indica L.

possesses antioxidant activity and precludes carbon tetrachloride (CCl4)-

mediated oxidative hepatic damage in rats,” was also able to evaluate the

aqueous extract of Paragis to protect rats from hepatic-injuries with the use of

its many properties.

An article from Evidence-Based Complementary and Alternative

Medicine entitled “ Eleusine indica Possesses Antioxidant, Antibacterial, and

Cytotoxic Properties” (Al-Zubairi et al., 2011), was also able to give an

emphasis on the therapeutic benefits of medicinal plants, how the plant

“Paragis” was a component of basic remedy in many traditional medicines,

and how further investigation will lead to Paragis’s inclusion in general

medicine practice.

Justification of Proposed Study

Overall, the study aims to utilize Paragis or E. indica as an antibacterial

extract and test its efficiency on S. aureus. It is an attempt of the researchers

to scientifically prove that Paragis extract can be used against S.aureus. The

research was conducted by means of experimentation and cross-analysis of

its advantages and disadvantages as an antibacterial extract.

18
 CHAPTER III

METHODOLOGY

A. Research Design

An experimental method was used in this study, designed by the

students with the help of qualified professionals from Laguna State

Polytechnic University – Los Banos, to determine the efficiency of the

Paragis (E. indica) as an antibacterial extract against S. aureus.

B. Research Locale

This whole process of this study was conducted at different

institutions. The collection of the Paragis (E. indica) sample was

conducted at the field beside Muntinlupa Sports Complex, Muntinlupa

City. The Ethanol Extraction was conducted at the Adamson University

Technology Research and Development Center (AUTRDC), and the

experimentation process was held at the Laguna State Polytechnic

University – Los Banos (LSPU – LB).

Figure 3.1 Laguna State Polytechnic University – Los Banos

19
 C. Materials and Instrumentation

This study made us of the following materials : Paragis plant,

Ethanol Soxhlet Extraction Set-up (AUTRDC), test tubes, pipettes, 3

petri dishes, Staphylococcus aureus strain, 9 paper disks, 3 cotton

buds, and incubator.

D. Methods

a. Gathering

b. Extraction

c. Cultivation of Bacteria

d. Kirby-Bauer Antibiotic Susceptibility Test

Procedures

The researchers first collected the Paragis (E. indica) plant at the field

beside Muntinlupa Sports Complex, as the Paragis plant can be found almost

anywhere. After collecting the Paragis plant, it was taken to the Adamson

University Technology Research and Development Center (AUTRDC) for the

Ethanol Extraction Process. The researchers also scheduled a date for the

experimentation at the Laguna State Polytechnic University – Los Banos.

After the ethanol extract of Paragis was taken back at AUTRDC, only

then did the experimentation proper started. The Kirby-Bauer Antibiotic

Sensitivity Test was conducted by the researchers, along with qualified

professionals from Laguna State Polytechnic University – Los Banos (LSPU –

LB). Due to the holiday, the experimentation proper was postponed. The S.

20
aureus was revived by a research consultant from LSPU – LB. 9 paper discs

were impregnated with Streptomycin (positive control), Distilled water

(negative control), and Paragis extract – 75% pure (experimental group), and

left for 3 hours. After that, the set-ups were prepared. For the

experimentation, there were 3 set-ups, each of it with 3 groups: 1 positive

control group (Streptomycin); 1 negative control group (distilled water); and 1

experimental group (Paragis extract – 75% pure). For each set-up. An agar

plate was prepared, and each was divided into 3, wherein one division was for

one specific group. The same was done to the other set-ups. Before the

researchers placed the paper discs into its respective zone, the agar plate

was streaked with the bacterial strain S. aureus. After the streaking and the

placing of the paper discs in the agar plates. The agar plates were then

placed in an incubator, set at 37° C for 24 hours. The data was gathered after

the 24 hours of incubation.

Data Gathering Procedure

The data was gathered mainly by measuring the zone of inhibition

yielded by the S. aureus as a response to the antibacterial activities of each of

the groups (positive control group – Streptomycin; negative control group –

distilled water; and experimental group – Paragis 75% pure extract.)

Statistical Treatment

This study has a quantitative data to be measured, namely the zone of

inhibition. The zone of inhibition , shown in millimetres (mm), was measured

after 24 hours of incubation at 37° C for each of the groups at the 3 different

set-ups.

21
 CHAPTER IV

RESULTS AND DISCUSSION

Results

Comparison of susceptibility tests to measure the ability to inhibit

bacterial growth among Positive Control (Streptomycin), Negative

Control (Distilled water), and Experimental Control (Paragis Extract)

within three trials

The bacterial colonies were identified by its dirty white color on the agar

plates.

Figure 4.1 Kirby- Bauer Susceptibility Test to Measure Antibiotic Sensitivity

The Kirby-Bauer (also called disk diffusion test) is widely used in the

field of research and known as a standard susceptibility test. In this method of

testing, the bacteria and other agents are placed on a plate of solid growth

medium and wafers of antibiotics (white disks, shown) are added to the plate.

22
After allowing the bacteria to grow overnight (for 24 hours), areas of media

surrounding the disks indicate that the agent inhibit bacterial growth.

As shown in the image above, only the positive control inhibits bacterial

growth among the three control groups while the other two groups remain

bacterial free zones.

Zone of Inhibtion Bacterial Activity

Trial 1 Trial 2 Trial 3

Positive Control
25mm 26mm 29mm Susceptible
(Streptomycin)

Negative Control
0mm 0mm 0mm Resistant
(Distilled Water)

Experimental
Control 0mm 0mm 0mm Resistant
(Paragis Extract)

Discussion

According to our table, we can see that the positive control

(Streptomycin) has a 25mm zone of inhibition for the 1st trial, 26mm zone of

inhibition for the 2nd trial, and 29mm zone of inhibition for the 3rd trial. The

zone of inhibition for the Negative Control (distilled water) and Experimental

Control (Paragis -75% pure extract) is all zero for every trial. The S. aureus is

susceptible to the positive control but resistant to the negative control and

experimental control.

23
 CHAPTER V

SUMMARY, CONCLUSION, AND RECOMMENDATIONS

Summary

The first few weeks were all about the proposal & constructing of the

research plan. Fortunately, our study has been successfully approved for the

first time we proposed. The following weeks is spent by making an

appointment for our experimentations as well as the collection of the materials

& equipments. Over the succeeding weeks things went well, we were able to

submit the sample Paragis to the Adamson University Technology Research

and Development Center (AUTRDC) & convey the request to the Laguna

State Polytechnic University Los Baños Campus. The extraction lasts for

about 2 weeks while the experimentation proper was conducted over the span

of 4 weeks. Our experimentation consists of three controls with three trials.

Using the method of Kirby-Bauer test, we found out the efficiency of the

Paragis extract.

Conclusion

As a conclusion, the overall results would be able to let the people gain

further knowledge on what they can do to improve or utilize the Paragis (E.

indica) extract and its properties, be it antibacterial or not, as the results can

be used as a platform for scientific discoveries. The Paragis (E. indica) was

inefficient as an antibacterial extract against S. aureus, as shown in the

results garnered from the experimentation proper.

Recommendation

24
 For further studies, the researchers recommend finding another

alternative extraction method that might be able to efficiently extract the

properties of Paragis (E. indica) that makes it antibacterial. The y may also try

to use it on other bacterial strains other than S. aureus to see if it would have

an antagonistic effect on the bacterial strains. The researchers also

recommend experimenting with larger concentrations of Paragis (E. indica).

25
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Dr. Agustin, F (2018). Antimicrobial Activity of Paragis. Retrieved from

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27