Antibacterial Activity of Niyog-niyog Phyllanthus niruri Fruit Ethanolic Extract against

Escherichia coli

A RESEARCH STUDY

Official Entry for

Division Scilympics 2019

Secondary Level

Life Science

Individual Category

Researcher:

Christian Angelo Bingco

Jeah Loraine Ortega

Research Adviser

Antique National School

San Jose, Antique

September 2019
 Antibacterial Activity of Niyog-niyog Phyllanthus niruri Fruit Ethanolic Extract against
Escherichia coli

Research Plan

A. Rationale

Bacteria are everywhere, whether in food, clothing, appliances and any things we touch

or contact with. It is one of the common and leading causes of death in the world. It is said that

humans play an important role in disease risk, infection patterns and bacterial distribution

(Frank, et. al., 2018). Escherichia coli is one of the most frequent causes of many common

bacterial infections, including cholecystitis, bacteremia, cholangitis, urinary tract infection (UTI),

and traveler's diarrhea, and other clinical infections such as neonatal meningitis and pneumonia.

It is usually transmitted through eating (Tarun Madappa, 2019). Most varieties of E. coli are

harmless or cause relatively brief diarrhea (Debra R. W., et al., 2017). The study focus more on

creating an alternative way using Niyog - niyog Phyllantus niruri fruit extract against bacterial

infections. Discovering and finding a new medicine against this bacterium is a big deal in the

society. Imagine eating food with bacteria manifesting it. This study will greatly impact not only

the society but also the medicinal world.

 B. Research questions, Hypotheses, Engineering goals, and Expected outcomes

1. Research questions

1.1 What are the active compounds found present in Niyog - niyog Phyllanthus niruri

fruit that can exhibit antibacterial activity?

1.2 What is the antibacterial activity of Niyog - niyog Phyllanthus niruri fruit ethanolic

extract against Escherichia coli at different concentration?

1.3 Is there a significant difference in antibacterial activity of Niyog - niyog Phyllanthus

niruri fruit at different concentrations?

2. Hypothesis

The following hypotheses will be put forward:

2.1 There is no active compounds found present in Niyog- niyog Phyllanthus niruri fruit

that can exhibit antibacterial activity

2.2 there is no antibacterial activity of Niyog –niyog Phyllanthus nirui fruit ethanolic

extract that can use against Escherichia coli.

2.3 there is no significant difference in the antibacterial activity of Niyog -niyog

Phyllanthus niruri at different concentrations.

C. Description in Details of Methods or Procedure

1. Procedure and Experimental design

The researchers will collect fifty (50) grams of Niyog – niyog Phyllanthus niruri Fruit

from the source days before the conduct of the study. The collected Fruits will be washed with

distilled water. After, the washed fruits will be weighed in a gram scale and will be pounded
using mortar and pestle. The pounded fruit will be put on a clean container and will be soaked

with 300ml ethanol for twenty – four (24) hours. Ethanol will be bought on the available

pharmacy. After the twenty – four (24) hours, the pounded fruit will be filtered over and over

until there are no remains/ residue on the extract. The researchers will also collect the

Escherichia coli from the laboratory of West Visayan State University, with the supervision of

certified medical technologist.

The researchers will prepare the apparatuses and equipment needed for the study such as:

one (1) strainer, two (2) clean containers, one (1) gram scale, two (2) pair of mortar and pestle

one (1) micropipette, twelve (12) petri dish, one (1) cork borer, four (4) beakers and twelve (12)

graduated cylinders. It will be prepared prior to the conduct of the study. The researchers will

also collect one (1) liter of ethanol from the source days before the conduct of the study.

Minutes after bringing, the researchers will separate fresh with the rotten ones. The

researchers will wash the collected fifty (50) grams of Niyog - niyog Phyllanthus niruri Fruit

with tap water. The researchers will extract the five (5) grams of powder with twenty-five (25)

ml of ethanol. After the process, it will be put on the rotary evaporator to recover excess solvents

to oily crude. The extract will be put on air-tight seal vials to prevent any contaminations.

The bacteria Escherichia coli will be prepared by the registered medical technologist.

The process will start by preparing petri dishes. The bacteria Escherichia coli will be put on the

separate petri dishes; the researcher apply the Niyog – niyog Phyllanthus niruri fruit ethanolic

extract based on the treatments. The plates will be left to stand for ten (10) minutes.

The experiment will use four (4) treatments with 3 replicates each.

The four (4) treatments will be used in the study will be:
Treatment A = 100 µl of pure (100%) Niyog - niyog Phyllanthus niruri fruit ethanolic extract

Treatment B = 100 µl of 50% (50µl) of Niyog – niyog Phyllanthus niruri fruit ethanolic extract

+ 50 µl Distilled water

Treatment C (positive control) = 100 µl of Ceftriaxone

Treatment D (negative control) = 10 µl Distilled water

2. Risk and Safety

The researchers used gloves, mask and laboratory gowns to prevent contact with bacteria

and hazardous chemicals. The researchers put the ethanol in a secured container to prevent

any mishaps.

3. Data analysis

Using Agar Well Diffusion Method, the researchers compared the different

concentrations to know the effectivity of each variables and Phytochemical Analysis to

support it.

4. Data gathering procedures

After the incubation the researchers will measure the inhibition using Vernier

Caliper and after recording, the data average of the inhibition will be computed using a

standard calculator. Phytochemical analysis will be done after measuring the average

inhibition. Analysis of variance will be executed using one (1) way or Single – factor

ANOVA.
 BIBLIOGRAPHY

A. Websites

Adeneye, AA.J., et al. (2012). Leaf and seed aqueous extract of Phyllanthus amarus
improves insulin resistance diabetes in experimental animal studies.Ikeja Lagos State, Nigeria
https://www.ncbi.nlm.nih.gov/pubmed/23085308

Odetola, A. A. and Akojenu, S. M. (2000) Anti-diarrhoeal and gastro-intestinal potentials of

the aqueous extract of Phyllanthus amarus (Euphorbiaceae). Ibadan, Nigeria.
https://scialert.net/abstract/?doi=rjphyto.2009.77.84

B. PDFs

Chandra, R: Lipid lowering activity of P. niruri, Journal of Medicinal & Aromatic Plant

Sciences. 22 (1): 29-30. (2010)

https://pdfs.semanticscholar.org/cb52/877e900305936c7f5d8f0c387ae4961e5371.pdf

Okoli CO, Ibiam AF Ezike AC et al. (2010) Evaluation of antidiabetic potentials of

Phyllanthus niruri in alloxan diabetic rats. African Journal of Biotechnology (2010); 248-259.

https://www.ajol.info/index.php/ajb/article/viewFile/77801/68223

Nelli G. P.., et al. (2014) Aqueous Extract of Phyllanthus niruri Leaves Displays in

Vitro Antioxidant Activity and Prevents the Elevation of Oxidative Stress in the Kidney of

Streptozotocin-Induced Diabetic Male Rats; Evidence-Based Complementary and Alternative

Medicine, (2014) 10 pp 43-50.

https://www.hindawi.com/journals/ecam/2014/834815/