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ANTIBACTERIAL EFFECT OF ALAGAW (PREMNA SERRATIFOLIA)

 

AGAINST STAPHYLOCOCCUS AUREUS

 

 

Thesis Outline Submitted to the Faculty

 

of the Department of Biology

 

Davao Medical School Foundation, Inc.

 

 

In Partial Fulfillment

 

of the Requirements for the Degree of

 

Bachelor of Science in Biology,

 

 

by

 

Name of Student A

 

Name of Student B

 

Name of Student C

 

 

September 2015

 

 

INTRODUCTION

 

 

Staphylococcus aureus is a type of bacteria found on the skin and mucous

 

membranes of humans. It is a gram positive bacterium that is round in shape and form

 

into clusters. S. aureus is one of the most important bacteria in human diseases (Stanway,

 

2014). It causes most of the staph infections where skin infections are the most common.

 

Staph aureus can look like pimples or boils. They may be red, swollen and painful, and

 

sometimes have pus or other drainage. They can turn into impetigo, which turns into

 

cellulitis (U.S. National Library of Medicine, 2014).

 

Premna serratifolia is a small tree that is 3 to 8 meters high and is indigenous in

 

the Philippines. Its leaves are broad, heart-shaped, covered with short hair-like structures,

 

and are 10 to 20 millimeters long. The flowers of alagaw are pale green in color and are 4

 

to 5 millimeters long. The fruit is green, fleshy, rounded, and about 5 millimeters in

 

diameter. Alagaw trees are distributed in many parts of the Philippines, specifically in

 

Batan Islands, northern Luzon, and in Mindanao (Stuart, 2014).

 

Alagaw is used in variety of ways and provide health benefits including culinary

 

uses, antiseptic, antioxidant, antibiotic properties, and even anti-parasitic usage (Lasco,

 

2011). The leaves of the Alagaw are used as treatment for vaginal irritation, fever and

 

colds, cough and bronchitis, stomachaches, and gas pains. Its leaves are also used in

 

bathing infants, treatment for beriberi, and ticks and fleas while the roots are chewed and

 

the saliva are swallowed for cardiac troubles (Neil, 2015).

 

A lot of people are using alternative medicines in finding cure for their illness

 

because it is cheaper than conventional medicines. Since Premna serratifolia is a native

 

 

plant in the Philippines, the outcome of this study will provide knowledge about the

 

antibacterial property of Alagaw against Staphyloccocus aureus for those communities

 

with easy access on this herbal medicinal plant.

 

The objective of this study is to determine the antibacterial properties of Alagaw.

 

The specific objective is to:

 

1. To determine if Staphylococcus aureus is susceptible to inhibition by Alagaw

 

extract.

 

 

Fig. 1. Premna Serratifolia Tree

 

 

Fig. 2. Leaves

 

 

Fig. 3. Fruits

 

 

REVIEW OF LITERATURE

 

 

Alagaw has been a subject of many studies to provide evidence-based support to

 

its therapeutic claims. One species of Alagaw, the Premna odorata, a component used in

 

a commercialized Philippine herbal preparation called ?Pito-pito? had been a focus of one

 

study conducted in Iligan City. They isolated flavones from P. odorata specifically,

 

flavone aglycones from the ethanolic crude extract of the leaves. These flavones were

 

said to exhibit antimicrobial, anti-inflammatory, and chemopreventive properties. Thus,

 

the study provided an evidence for the medicinal use of the P. odorata (Pinzon et al.,

 

2011).

 

Another species of Alagaw, the Premna serratifolia which is also known as

 

?Munney? or ?Agnimantha?, became a subject of another study that was conducted in

 

India. It was shown that this species of alagaw presented some antimicrobial activity. In

 

this study, extracts from the bark and wood of P. serratifolia was tested against nine

 

bacterial and four fungal organisms by disc diffusion method. The results of the study

 

revealed that all the extracts possessed a broad scale of antimicrobial activity (Sathak et

 

al., 2010).

 

In another study conducted in India by Rajendran and Basha (2010), the P.

 

serratifolia was screened to evaluate in vitro antimicrobial activity against some human

 

pathogenic organisms. Findings of their study showed potential antimicrobial properties

 

of the extracts and fractions from the root of P. serratifolia against different bacterial

 

organisms and fungal tested, and the zone of inhibition of the extracts and fraction were

 

found to be similar with that of standard antibiotics like ciproflaxin and ketoconazole.

 

 

The antibacterial action of the extract are said to be possibly due to the presence of

 

phytoconstituents such as, alkaloids, steroids, flavonoids, phenolic compounds, and

 

glycosides specifically iridoid glycosides.

 

Other studies were also conducted that revealed anti-viral and anti-parasitic

 

activity of alagaw species. In Malaysia, 61 medicinal plants were screened for antiviral

 

and cytotoxic activities where antiviral activity was tested against the herpes simplex

 

type-1 (HSV-1) and vesicular stomatitis viruses (VSV). One of the medicinal plants used

 

in the study was the P. odorata and the results have shown that the Premna odorata have

 

selective activity against VSV (Ali et al., 1996). In New Caledonia, 29 extracts of 18

 

medicinal plants were evaluated against various parasites, such as Leishmania donovani,

 

Trypanosoma brucei, Trichomonas vaginalis, and Caenorhabditis elegans. The P.

 

serratifolia was among the most active against Leishmania donovani (Desrivot et al.,

 

2007).

 

 

MATERIALS AND METHODS

 

Place and Time of the Study

 

The leaves sample of alagaw will be collected from Sto. Nino, Mintal, Davao City

 

and the experiment will be conducted at Davao Medical School Foundation Main

 

Laboratory.

 

 

Fig. 4. Sto. Nino, Mintal, Davao City

 

 

Fig. 5. DMSF

 

 

Preparation of Sample Plant and Alagaw Extract

 

Preparation of alagaw leaves by ethanol extraction (Hoque et al., 2011):

 

1. The Alagaw leaves will be washed in distilled water and cut into small pieces.

 

2. Then the alagaw leaves will be dried at 40?C for 48 hours in an oven.

 

3. After the alagaw leaves have dried, it will then be grounded into fine powder

 

using a mortar and pestle.

 

4. Weigh 20 grams of Alagaw leaf powder and soak it in 80ml of 95% ethanol in

 

sterilized bottle and keep it in fume hood chamber for overnight.

 

5. The ethanol fraction will be separated by using sterilized cheesecloth and filtered

 

through sterilized Whatman filter paper (No.3).

 

Antimicrobial Test

 

The antimicrobial activity of Alagaw leaf extracts will be tested by disk diffusion

 

method (Mukhtar, 2012). The filter paper disc will be prepared and sterilized. The S.

 

aureus will be transferred from nutrient broth to the Muller Hinton agar plates with the

 

help of a sterilize cotton swab and then the discs will be placed over the agar plates.

 

Then, the alagaw leaf extracts will be transferred to each disc. The plates will then be

 

incubated in an upright position at 37?C for 24 hours. After 24 hrs, the diameter of zone

 

of inhibition will be measured in millimeter and the results will be recorded.

 

Statistical Analysis

 

The experiment will involve four treatments, T1- 10%, T2- 25%, T3- 50%

 

concentration of extract, and T4- water (as the control), arranged in a COMPLETELY

 

RANDOMIZED DESIGN (CRD) using five plates per treatments replicated three times.

 

Dummy Table 1. Raw data on the Zone of Inhibition of extracts against S. aureus.

 

Concentration of extract

 

 

10%

 

 

25%

 

 

50%

 

 

Control

 

(Water)

 

 

Trial

 

1

 

2

 

n

 

 

Dummy Table 2. Analysis of Variance on the Zone of Inhibition of extracts against S.

 

aureus.

 

Source of Variation

 

 

SS

 

 

df

 

 

Concentration of extract 1 (10%)

 

Concentration of extract 2 (25%)

 

Concentration of extract 3 (50%)

 

Concentration of extract 4 (water)

 

Total

 

 

LITERATURE CITED

 

 

MS

 

 

F

 

 

P-value

 

 

F crit

 

 

Ali, A., Mackeen, M., El-Sharkawy, S., Hamid, J., Ismail, N., Ahmad, F., & Lajis, N.

 

(1996) Antiviral and Cytotoxic Activities of Some Plants Used in Malaysian

 

Indigenous Medicine [Abstract]. Pertanika J. Trop. Agric. Sci, 19(2), 129-136.

 

Casella, G. (2008). Complete Randomized Design. In Statistical Design, 43-90. Retrieved

 

July 18, 2015, from http://www.springer.com/gp/book/9780387759647

 

Desrivot, J., Waikedre, J., Cabalion, P., Herrenknecht, C., Bories, C., Hocquemiller, R., &

 

Fournet, A. (2007). Antiparasitic activity of some New Caledonian medicinal

 

plants. Retrieved July 10, 2015, from

 

http://www.ncbi.nlm.nih.gov/pubmed/17329051

 

Hoque, M., Rattila, S., Asaduzzaman, M., Bari, L., Inatsu, Y., & Kawamoto, S. (2011) Antibacterial

 

Activity of Ethanol Extract of Betal leaf (Piper betle L.) Against some Food Borne Pathogens.

 

Bangladesh J Microbiol, 28(2), 58-63. Retrieved July 15, 2015, from

 

http://www.researchgate.net/publication/220004932_Antibacterial_Activity_of_Ethanol_Extra

 

ct_of_Betel_leaf_(Piper_betle_L.)_Against_some_Food_Borne_Pathogens

 

 

Lasco, G. (2011). HALAMANG GAMOT: ALAGAW. Retrieved July 10, 2015, from

 

http://kalusugan.ph/halamang-gamot-alagaw/

 

Neil. (2015). Alagaw Alternative Medicine Supplements and Vitamins. Retrieved on July

 

10, 2015, from

 

http://plantfruitstreesmedicineherbal.blogspot.com/2012/09/alagaw-alternativemedicine-supplements.html

 

Pinzon, L., Uy, M., Sze, K., Wang, M., & Chu, I. (2011). Isolation and characterization of

 

antimicrobial, anti-inflammatory and chemopreventive flavones from Premna

 

odorata Blanco. Journal of Medicinal Plants Research, 5(13), 2729-2735.

 

Retrieved July 20, 2015

 

 

fromhttp://www.researchgate.net/publication/266447476_Isolation_and_character

 

ization_of_antimicrobial_antiinflammatory_and_chemopreventive_flavones_from

 

_Premna_odorata_Blanco

 

Rajendran, R., & Basha, S. (2010). Antimicrobial activity of crude extracts and fractions

 

of Premna serratifolia Lin. root. Medicinal Plants, 2(1). Retrieved July 20, 2015,

 

from

 

http://www.academia.edu/1847834/Antimicrobial_activity_of_crude_extracts_and

 

_fractions_of_Premna_serratifolia_Lin._root

 

Sathak, M. (2010). Antimicrobial Activity of Different Bark and Wood of Premna

 

serratifolia Lin., International Journal of Pharma and Bio Sciences, 1(1).

 

Retrieved from http://www.ijpbs.net/21.pdf

 

Stanway, A. (2014). Staphylococcal skin infections. Retrieved July 10, 2015, from

 

http://dermnetnz.org/bacterial/staphylococci.html

 

Stuart, G. (2014). Alagaw. Retrieved on July 10, 2015, from

 

http://www.stuartxchange.org/Alagaw.html

 

U.S. National Library of Medicine. (2015). Staphylococcal Infections. Retrieved July 10,

 

2015, from http://www.nlm.nih.gov/medlineplus/staphylococcalinfections.html

 

 

2015

 

 

MONTH

 

ACTIVITY

 

 

J

 

 

Discussion of Possible Topic

 

Start of Thesis Outline Preparation and

 

Review of Literatures

 

Submission of Outline to Adviser

 

Outline Defense

 

Lab Experimentation & Data Gathering

 

Manuscript Preparation

 

Submission of Final Paper

 

Thesis Defense

 

Finalization of Paper

 

SCHEDULE OF ACTIVITIES (GANNT CHART)

 

 

BUDGETARY REQUIRMENT

 

 

J

 

 

A

 

 

S

 

 

O

 

 

# of travels

 

(round

 

trip)

 

 

# of

 

pax

 

 

Place to

 

visit

 

 

Transportation

 

 

Amount

 

 

Total

 

 

1

 

 

2

 

 

Mintal

 

 

Jeepney

 

 

20

 

 

40

 

 

2. Communication

 

& Electricity

 

 

Cost

 

 

# days

 

 

Laptop/Computer/

 

Telephone/

 

Internet/Cellphone

 

Bills

 

 

50

 

 

30

 

 

1,500

 

 

1,500

 

 

11

 

 

11

 

 

TRAVEL

 

1. Land

 

Transportation

 

 

3. Supplies and

 

Materials

 

 

qty

 

 

unit

 

 

Print

 

 

11

 

 

pages

 

 

GRAND TOTAL

 

 

1,551

 

 



 

---

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