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Antibacterial Performance of Fruit Extracts of Wild Variety of Trapa Quadrispinosa Roxb. Found in Bangladesh

Md. Anowar Razvy
Department of Agronomy and Agricultural Extension, University of Rajshahi, Rajshahi- 6205, Bangladesh

Shaikh Mizanur Rahman
Department of Genetic Engineering and Biotechnology, Jessore University of Science and Technology, Jessore, Bangladesh

Md. Motiur Rahman
Plant Breeding and Gene Engineering Lab., Department of Botany, University of Rajshahi, Rajshahi- 6205, Bangladesh

Md. Aminul Hoque
Department of Agronomy and Agricultural Extension, University of Rajshahi, Rajshahi- 6205, Bangladesh

Abstract/ Executive Summary:

Water chestnut (Trapa quadrispinosa Roxb.) is an annual aquatic herb fruit plant grown in fresh water basin of tropical, sub-tropical and some temperate part of the world. It is called “Singhara” in Bangladesh. The methanol extract of wild variety of water chestnut showed the highest antibacterial performance (zone of inhibition 34mm) against Bacillus megaterium with the doze of 200μg/disc. Whereas, the lowest antibacterial activities (10mm) were recorded for aqua’s extract and petroleum ether extract in comparison to standard antibiotics, kanamycin. Also all gram negative bacterial strains were recorded as resistance in 100μg/disc dose of aqua’s extract. In this disc diffusion assay, the methanol extract of wild variety was found to have a significant antibacterial effect than the other extracts which is in fact put forward the potency to develop a Phyto-medicine against microbes.

Keywords: Trapa, Wild, Fruit extract, Antibacterial activities

Location: Experimental field of Botanical garden at Rajshahi University campus, Rajshahi

Start Date: 00-09-2010

End Date: 00-09-2010

Program Area: Development of Host and Medicinal Plants

Commodity: Water chestnut

Objectives:

To find out antimicrobial activities of fruit extracts of water chestnut.

Methodology:

Plant materials - Plant materials were used from mature fresh fruit of wild water chestnut (T. quadrispinosa) collected from the experimental field of Botanical garden at Rajshahi University campus, Rajshahi- 6205, Bangladesh during September 2010 which was originally collected from Naogaon, Bangladesh. Microbial strain - Eight pathogenic bacteria where five strains of gram-positive such as Staphylococcus aureus, Bacillus subtilis, Bacillus megaterium, Sarcina lutea and Bacillus cereus and three strains of gram-negative for instance, Escherichia coli, Salmonella typhi and Shigella sonnei were used for bioassay study. The pure strains were identified and obtained from pathology laboratory, Department of Botany, University of Rajshahi, Rajshahi- 6205, Bangladesh. Microbial culture media - To evaluate antibacterial activity and sub culturing of test organisms, nutrient agar media having 0.5gm peptone, 1gm yeast extract, 0.5gm sodium chloride and 2gm agar were dissolved in distilled water. This composition of the nutrient was maintained throughout the experiment. Preparation of fresh culture - The nutrient agar medium was prepared and dispersed (5ml in each test tube) in clean test tubes to get ready slants. The test tubes were sterilized for 30 minutes and kept in an inclined position (45^oC) for solidification. The test organisms were transferred to the agar slants from the supplied pure cultures with the help of an inoculating loop in an aseptic condition. The inoculated slants were then incubated at 37.5oC for 24 hours to assure the growth of test organisms. These fresh cultures were used for the sensitivity test. Preparation of the test plates - Nutrient agar media were poured approximately 4mm in each petridish in aseptic area and petridishes were rotated several times, first clockwise and then anticlockwise. Then 200μl of cultured test organism in nutrient broth media was spreaded on the surface of solid nutrient agar media, and preserved for inoculation of sample and standard discs. Preparation of test sample - Different amount of extracts such as 5mg, 10mg and 20mg of aqua’s extract (AE), methanol extracts (ME) and petroleum extract (PE) of water chestnut sample (wild variety) were dissolved in 0.5ml water in separate glass vial. Thus, for each extract the concentrations were 100μg/μl, 150μg/μl and 200μg/μl, respectively. Preparation of discs - Three types of discs were prepared for antibacterial screening. Sample discs - Sterilized (BBL) filter paper discs (5mm in diameter) were prepared with the help of punch machine and were taken in an empty petridish. Sample solution of desired concentration (10μl/disc) was applied on the discs with the help of a micropipette in an aseptic condition. Standard discs - Standard discs were used to compare the antibacterial activity of test material. In our investigation, kanamycin (100μg/disc) was used as standard disc. Solvent control discs - Solvent control discs were prepared using filter paper (5mm) and same volume of residual solvent without sample and these were then used as negative control to ensure the residual solvent. Placement of the discs and incubation - The dried crude extract discs and standard discs were placed gently as 20mm apart from each other and 15mm outlying the edge of the plate to avoid overlapping the zone of inhibition on the solidified agar plates seeded with the test organisms. The plates were kept in a refrigerator at 4oC for 24 hours in order to provide sufficient time to diffuse the antibiotics into the medium. Then the plates were incubated at 37.5^oC for 24 hours in an incubator. Measurement of the zone of inhibition - After incubation, the antibacterial activities of the test samples were determined by measuring the diameter of inhibitory zones in term of millimeter (mm) with a transparent scale.

Source of Information: International Journal of Biosciences (IJB); ISSN: 2220-6655 (Print) 2222-5234 (Online); Vol. 2, No. 11, p. 178-183, 2012

Article Link (Online Journal): http://www.innspub.net

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The study demonstrated that herbal medicine could be as effective as modern medicine to combat pathogenic microorganisms. This work has highlighted the antimicrobial effects of fruit of T. bispinosa on some of the medically important pathogens. This is in fact a promising result, with the comparable standard antibiotics and suggests the potency of these extracts. Hence, T. bispinosa fruit could be used as a guide in our continuing search for being used in the field of biopharmaceutical industry or in ethno-medicine.

Knowledge Management: Journal