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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,
Name of Student A
Name of Student B
Name of Student C
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
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.,
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
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.,
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
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).
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.
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
Dummy Table 2. Analysis of Variance on the Zone of Inhibition of extracts against S.
Source of Variation
Concentration of extract 1 (10%)
Concentration of extract 2 (25%)
Concentration of extract 3 (50%)
Concentration of extract 4 (water)
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
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
Lasco, G. (2011). HALAMANG GAMOT: ALAGAW. Retrieved July 10, 2015, from
Neil. (2015). Alagaw Alternative Medicine Supplements and Vitamins. Retrieved on July
10, 2015, from
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
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,
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
Stuart, G. (2014). Alagaw. Retrieved on July 10, 2015, from
U.S. National Library of Medicine. (2015). Staphylococcal Infections. Retrieved July 10,
2015, from http://www.nlm.nih.gov/medlineplus/staphylococcalinfections.html
Discussion of Possible Topic
Start of Thesis Outline Preparation and
Review of Literatures
Submission of Outline to Adviser
Lab Experimentation & Data Gathering
Submission of Final Paper
Finalization of Paper
SCHEDULE OF ACTIVITIES (GANNT CHART)
# of travels
3. Supplies and
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