Sip - Ranhs

Microbial Activity of Crude Extract of Tetrastigma hermandii (Alupidan) against S.
aureus, E. Coli and C. albicans
A Science Investigatory Project of Ramon Avanceña National High School
Statement of the Problem

This study is aims to determine the microbial activity of Alupidan (Tetrastigma hermandii) against
Staphylococcus aureus, Escherichia Coli and Candina albicans.
Specifically, this project seeks to answer the following question, “Is there a significant difference in the zone of
inhibitions of the different concentrations of Tetrastigma hermandii and commonly used anti-microbial agents
againt S. aureus, E. coli and C. albicans?”
Introduction
Medicinal plants have a long history of use and their use is widespread in both developing and developed
countries. According to reports of the World Health Organization, 80% of the world’s population relies mainly
on traditional therapies which involve the use of plant extracts or their active substances (WHO, 1993).
Microorganisms have developed resistance against many antibiotics due to the indiscriminate use of
antimicrobial drugs (Ahmad et al., 1998). Furthermore, antibiotics are sometimes associated with side effects
(Cunha, 2001), whereas there are some advantages of using antimicrobial compounds of medicinal plants,
such as fewer side effects, better patient tolerance, relatively less expensive, acceptance due to long history of
use and being renewable in nature (Vermani and Garg, 2002).
Tetrastigma hermandii, locally known as “Alupidan” in the Visayas, is a member of the Vitaceae genus which is
comprised by approximately 95 species, distributed in tropical and subtropical Asia (Planchon 1887; Latiff
1983; Chen et al. 2011; Trias-Blasi et al. 2012; Wen 2007). Most of the members of the Vitaceae family show
microbial activity (Lui et al, 2003), aricidal activity (Adarsh et al, 2013), antiviral activity (Yang, 1989)have
phenolics, flavonoids contents and have antioxidant activity (Hossain et al, 2011).
Alupidan is woody vine, that has sour fruits and leaves that usually used in cooking to add flavoring and is
suitable for making preservatives. According to traditional folklore, the decoction of the plant was used as a
powerful diuretic and externally as a lotion to treat scabies. (Brown, 1957)
Hypothesis
There is a significant difference in the zone of inhibitions of the different concentrations of Tetrastigma
Hermandii and commonly used anti-microbial agents against S. aureus, E. coli and C. albicans.
Materials
Nutrient Broth, Nutrient Agar, 95% ethanol, distilled water, Petri plates, glass spreader, wire loop, test tubes
and test tube rack, erlenmeyer flasks, mortar and pestle, filter paper, incubator, autoclave, centrifuge
Procedure
Place of work
The present study entitled “Microbial Activity of Crude Extract of Tetrastigma Hermandii
(Alupidan) against S. aureus, E. Coli and C. albicans” was carried out in the Central Science Laboratory
of West Visayas State University.
Test microorganisms
The pathogenic organisms were obtained from
1. Preparation of the culture broth

Nutrient broth was prepared and was inoculated with the test organism. A loop full of
microorganism was taken and inoculated in the NB and was incubated at 37°C for 24 hrs to obtain a
viscous growth.
2. Preparation of plant extracts

The fresh samples of Tetrastigma hermandii (Alupidan) used in the present study were obtained
from a household at Brgy. Sto. Domingo, Arevalo and was identified by comparing it to a herbarium of
T. hermandii by Robinson (1912).They were collected and washed with distilled water. Leaves were
then crushed with distilled water using mortar and pestle. The same process was done with the stems.
The samples was then poured in sterilized centrifuge tubes and centrifuged at 3000 rpm for 15 min.
The supernatant was taken and different concentrations (1000, 1500, and 2000 ppm) were made using
sterile distilled water.
3. Preparation of Nutrient agar plates

The freshly prepared and autoclaved NA media was poured in the Petri plates, after cooling it to
45°C, and was kept to solidify. Cotton swabs were dipped in the culture broth and were swabbed on the
solidified media surface.
4. Preparation of Positive and Negative Control

The positive control for the experiment will be Amoxicillin for S. aureus and E. coli and
Clotrimazole for C. albicans while the negative control will be distilled water.
5. Antimicrobial sensitivity test using filter paper method

Filter paper discs of 5 mm diameter were prepared and sterilized by dipping them in 95%
ethanol using sterile forceps. These discs were dipped aseptically in respective extracts of appropriate
concentration and placed over NA plates seeded with respective microbes, with the help of sterile glass
spreader. The plates were incubated in an upright position at 37°C for 24h. The diameter of inhibition
zones formed was measured in mm and the results were recorded and compared to the positive and
negative controls.
Data List
Risk Assessment
Bibliography
● Latiff A. 1983. Studies in Malesian Vitaceae VII. The genus Tetrastigma in the Malay Peninsula.
Gardens’ Bulletin Singapore 36 (2): 213–228.
● Planchon J.E. 1887. Monographie des Ampélidées vrais. In: de Candolle A.F.P.P. & de Candolle C.
(eds) Monographiae Phanaerogamarum. Vol. 5, part 2: 305–654. G. Masson, Paris.
● Chen P., Chen L. & Wen J. 2011. The fi rst phylogenetic analysis of Tetrastigma (Miq.) Planch, the host
of Raffl esiaceae. Taxon 60 (2): 499–512.
● Trias-Blasi A., Parnell J.A.N. & Hodkinson T. 2012. Multi-gene region phylogenetic analysis of the
grape family (Vitaceae). Systematic Botany 37 (4): 941–950.
http://dx.doi.org/10.1600/036364412X656437
● Wen J. 2007. Vitaceae. In: Kubitzki K. (ed.) The Families and Genera of Vascular Plants. Vol. 9: 467–
479. Springer-Verlag, Berlin.
● Ahmad I, Memood Z, Mohammad F (1998). Screening of some Indian medicinal plants for their
antimicrobial properties. J. Ethnopharmacol. 62:183-193.
● World Health Organization (WHO) (1993). Summary of WHO guidelines for the assessment of herbal
medines. Herbal Gram 28:13-14.
● Cunha BA (2001). Antibiotic side effects. Med. Clinics North Am. 85:149-185.
● Vermani K, Garg S (2002). Herbal Medicines for Sexually Transmitted Diseases and AIDS. J.
Ethnopharmacol. 80:49-66.
● Hossain MA, Shah MD, Gnanaraj C, Iqbal M. In vitro total phenolics, flavonoids contents and
antioxidant activity of essential oil, various organic extracts from the leaves of tropical medicinal plant
Tetrastigma from Sabah. Asian Pacific Journal of Tropical Medicine 2011; 717-721.
● Yang X. L., Luo J. and Sun S. B. Researches on anti-virus function of Tetrastigma hemsleyanum. Hubei
Traditional Chinese Medicine, 1989; 4:40− 41.
● T. P. Adarsh Krishna, T. P. Ajeesh Krishna, V. N. Sanyo Raj et al., “Evaluation of phytochemical
constituents and proximate contents in ethanolic leaf extract of Tetrastigma leucostaphylum (Dennst.)
Alstone (Vitaceae) found in Western Ghats of Kerala, India,” Research Journal of Pharmaceutical
Sciences, vol. 2, no. 10, pp. 1–6, 2013
● D. Liu, J. H. Ju, and J. S. Yang, “Studies on chemical constituents from Tetrastigma hypoglaucum,”
Chinese Traditional and Herbal Drugs, vol. 34, pp. 4–6, 2003.
● Brown, W.H., 1951-1957. Useful plantsof the Philippines. Reprint of the 1941-1943 ed. 3 Volumes.
Department of Argriculture and Natural Resources. Technical Bulliten 10. Bureau of Printing, Manila,
Philippines.
● Robinson, C. B., 1912. Bureau of Science No. 18043, US National Herbarium Sheet 902954, Barcode
01150894

Sip - Ranhs

Uploaded by

Copyright:

Available Formats

Sip - Ranhs

Uploaded by

Document Information

Original Description:

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Sip - Ranhs

Uploaded by

Copyright:

Available Formats

Microbial Activity of Crude Extract of Tetrastigma hermandii (Alupidan) against S.

aureus, E. Coli and C. albicans

A Science Investigatory Project of Ramon Avanceña National High School

Statement of the Problem

1. Preparation of the culture broth

2. Preparation of plant extracts

3. Preparation of Nutrient agar plates

4. Preparation of Positive and Negative Control

5. Antimicrobial sensitivity test using filter paper method

You might also like