Sweet Potato (Ipomea batatas) Leaves Extract Against

Staphylococcus Aureus
Research Plan
A. Question or Problem being addressed
S. aureus has long been has been recognized as one of the most common bacteria that cause
disease and serious problems among children and adults (Minnesota Department of health,
February, 2010).It is the leading cause of skin and soft tissue infections such as abscesses
(boils),furuncles, and cellulitis. Although most staph infections are not serious, S. aureus can
cause serious infection such as bloodstream infections, pneumonia, or bone and joint infections.
The problem being addressed is to help to eliminate the S. aureus and reduces the risk of the
bacteria spreading either to other sites on the patients body, where they might cause infection, or
to other patients (Dr. Alan Johnson, 1960).

B. Goals/ Expected Outcomes/ Hypotheses

This study aim to test the antibacterial property of sweet potato leaves, a common plant in
the Philippines. The extracts is test on S. aureus, the most common bacteria that cause serious
infections. A commercial antibiotic is use to extracts from sweet potato leaves in terms of
zones of inhibition to S. aureus.
C. Procedures
Collection

Mylyn Grace Cabanalan, Grade 10- Special Science Class

Sweet potato leaves were collected in the morning to ensure the active photosynthesis

and cells activities took place.

Culture slant of microbes (Staphylococcus aureus) will be prepared by the DOST in
Lapaz, Iloilo City.

Extraction of sweet potato leaves

Use 10 grams of Clorox and 90ml. of water to disinfect leaves. The collected leaves of
the sweet potato will be extracted inside the laboratory to avoid contamination. Then
the crude extract of the leaves should place in a clean test tube to ensure its safety.

Preparation of treatments

Sweet potato leaves extract; pure culture of Staphylococcus aureus is prepared in the
laboratory; a 100 mL graduated cylinder used to measure the extract; and test tubes served
as the container of the extracts and inoculated bacteria; petri plates were prepared for the
test; nutrient agar (NA) and Nutrient Broth (NB) were used for the culture of the bacteria;
and filter paper for seeding the extract and the antibiotic.

Preparation of Culture Medium

The microbiological laboratory will be prepared by the in charge and the needed materials

will be provided by the institution.

Nutrient Agar (NA) and Nutrient Broth (NB) were used for the culture of bacteria and

filter paper for seeding the extract and the antibiotic.

Approximately 38 grams of Mueller-Hinton Agar will be diluted in 300 mL of distilled
water in Erlenmeyer flask and will be autoclaved for 15 minutes at 1210C to avail the ideal
amount of the medium dispensed in 15 Petri dishes.

Mylyn Grace Cabanalan, Grade 10- Special Science Class

The melted agar will be allowed to cool at 600C before dispensing unto the plates.

Preparation of the Bacterial Inocula

Culture slant of microbe (Staphylococcus aureus) will prepared by the DOST in Lapaz,
Iloilo City.

Prepare and dispense the nutrient broth concentration in 5 test tubes.

A loopful of bacteria from the culture slant will be diluted in the designated nutrient broth.

Slowly swirl it to mix.

To test the antibacterial activity, sweet potato leaves crude extract will be used.

The antibacterial activity will be studied by agar disc diffusion method inside a sterilized
laminar hood.

Microbiological Assay

The microbiological assay will be conducted in sterile laminar hood of University of the
Philippines Visayas Microbiology Laboratory.

The alcohol lamp will be lighted to keep the sterility of the area.

The bacterial inoculum will uniformly spread using sterile cotton swab on a sterile Petri
dish Mueller-Hinton agar in a zigzag manner three times beside a lighted alcohol lamp
inside the laminar flow.

In vitro antibacterial activity test will be carried out by disc diffusion method.

After all the plates will be swabbed with the bacteria, the discs will be put on the agar on
the designated points.

To test the antibacterial activity, Sweet potato leaves extracts using various solvent will be
dispensed in disc at 10 uL in a clockwise manner.

Mylyn Grace Cabanalan, Grade 10- Special Science Class

After all the treatments will be dispensed the controls, the Petri plates will be placed inside
then incubator for 18 hours at 3500C temperature.

After an overnight incubation, the Petri plates will be inspected for structure of zones of
inhibition around the filter paper discs.

Disposal

The used gel-liked agars will be scraped from the Petri dishes and disposed in
autoclavable plastics (Scoville, H, 2012).

All glasswares will be autoclaved and the workplace will be cleaned with Lysol solution
and will be sterilized by UV light for 30 minutes.

Data and Data Gathering Procedures

Results will be based on the diameter of the zone of inhibitions of test treatments and the
controls. Antimicrobial index will be determined to interpret the extracts activities.

By comparing the area of zone of inhibition of test will be standard concentration and
potency of test samples will be determined. (HubPages, 2013)

Inhibition of the bacterial growth will be measured in millimeters.

Diameter of Zone Inhibition

The diameter of zones will be measured by means of a ruler and the average data will be

taken from the data of the x and y axis of the zone.

The data will be plotted in tabulated form and treated statistically to determine the
antimicrobial percent present in sweet potato leaves against Staphylococcus aureus.

Statistical Data Analysis Procedure

Mylyn Grace Cabanalan, Grade 10- Special Science Class

The data obtained from the study will be subjected to the following descriptive and
inferential statistical treatment using the Statistical Package for Social Sciences (SPSS)
Software.

The statistical tools used in this study are:

Mean- the mean will be used to determine the average scores of the results of the treatments in
this study.
Standard Deviation- to determine the dispersion between the mean.
ANOVA- will be used to determine the difference of two or more means set at 0.05 level of
significance.
Duncans Multiple Range Test (DMRT)- to test the significance of the F-ratio obtained in the
study.

Mylyn Grace Cabanalan, Grade 10- Special Science Class

D. Biblioraphy

Pike, R. M. (1976). Laboratory-associated infections: summary and analysis of 3921

cases. Health Laboratory Science. Retrieved last January 26, 2015.

Baird-Parker, A. C. (1963). A classification of micrococci and staphylococci based on

physiological and chemical tests. J. Gen. Microbiol. 30:409-427. Retrieved last January
28, 2015.

von Eiff, C., R. R. Reinert, M. Kresken, J. Brauers, D. Hafner, and G. Peters for the
Multicenter Study on Antibiotic Resistance in Staphylococci and Other Gram-Positive
Cocci Study (MARS) Group. (2000). Nationwide German multicenter study on
prevalence of antibiotic resistance in staphylococcal bloodstream isolates and
comparative in vitro activities of quinupristin-dalfopristin. J. Clin. Retrieved last January
28, 2015.

Bauer, A. W., Kirby, W.M. M. and Sherris, J. C. (1966). Antibiotic susceptibility testing
by a single disc method. AM. J. Pathol. Retrieved last January 28, 2015.

National Committee for Clinical Laboratory Standards (NCCLS) (2000). Methods for
dilution antimicrobial susceptibility tests for bacteria that grows aerobically. Retrieved
last January 28, 2015.

Mylyn Grace Cabanalan, Grade 10- Special Science Class

Orrett, F.A. and Land, M. (2006). Methicillin resistant Staphylococcus aureus prevalence:
Current susceptibility pattern in Trinidad. BMC Infectious diseases. Retrieved last
January 28, 2015.

http://www.edu-sciece.com/2012/08/evaluation-of-antibacterialproperties.html

Mylyn Grace Cabanalan, Grade 10- Special Science Class