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In Vitro Antibacterial Effect of Tinospora Cordifolia Extracts Against Some Selective Bacterial Pathogens

Md. Shamim Hossain
Department of Biotechnology and Genetic Engineering, Faculty of Applied Science and Technology, Islamic University, Kushtia-7003, Bangladesh

Mst. Touhida khatun
Department of Biotechnology and Genetic Engineering, Faculty of Applied Science and Technology, Islamic University, Kushtia-7003, Bangladesh

Md. Mehedi Hassan
Department of Biotechnology and Genetic Engineering, Faculty of Applied Science and Technology, Islamic University, Kushtia-7003, Bangladesh; Department of Microbiology and Genetics and Institute for Medical Science, Chonbuk National University Medical School, Jeonju 561-712, South Korea

Abstract/ Executive Summary:

The petroleum spirit, methanol, dichloromethane and ethyl acetate extracts of the leaves of Tinospora cordifolia (Menispermaceae) were screened for their anti-microbial activity using disc-diffusion method on nutrient agar medium. This plant was tested against four bacteria; two Gram-positive bacteria (Bacillus subtilis and Sarcina lutea) and two Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae).All the organic solvent extract showed susceptibility against Sarcina lutea, E.coli and Bacillus subtilis whereas Klebsiella pneumoniae showed resistant against all the organic solvent extract. It was found that the antimicrobial activity of the methanol extract (11mm) showed the maximum zone of inhibition against Escherichia coli. On the other hand petroleum spirit extract showed the maximum inhibition against Bacillus subtilis (3mm) respectively. The minimum inhibitory concentration for petroleum spirit and dichloromethane extracts of Tinospora cordifolia were ranged between 32-512μg/mL for tested bacteria. The result of this study demonstrates the potentiality of Tinospora cordifolia as a source of antimicrobials that could be harness for use in the health care delivery process.

Keywords: Tinospora cordifolia, Medicinal plant, Antibacterial activity

Location: Department of Biotechnology and Genetic Engineering, Faculty of Applied Science and Technology, Islamic University, Kushtia

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Diseases and insects

Objectives:

The present study was an attempt to investigate the antibacterial activity of Tinospora cordifolialeaf extracts against gram positive bacteria Bacillus subtilis, Sarcinalutea, and gram negative bacteria Escherichia coli, Klebsiella pneumonia.

Methodology:

Collection of plant leaves - Healthy, disease free and mature Tinospora cordifolia plant was collected directly from Islamic university campus, Kushtia, Bangladesh. This plant was then botanically identified and name of the plant, place and date of the collection were recorded. Preparation of T. cordifolia leaves extracts - After collection of the T. cordifolia plant leaves was washed by running tap water, cleaned and then dried without sun. Then it was pulverized into a fine powder. Then 10 gm of the leaves powder was weighed with electric balance and 40 ml each of the solvent (methanol, petroleum spirit, ethyl acetate and dichloromethane) was added in each conical flask. The powder was extracted separately with Whitman No. 1 filter paper and the solvents were recovered by rotary evaporator. Tested bacteria - Antibacterial activity of Tinospora cordifolia leaves extracts were investigated against two Gram-positive (Bacillus subtilisand, Sarcina lutea) and two Gram-negative (Escherichia coli and Klebsiella pneumoniae) bacterial isolates obtained from the Microbial Type Culture Collection (MTCC) of Microbiology Laboratory of the Biotechnology and Genetic Engineering Department, Islamic University Kushtia, Bangladesh. The tested bacteria were culture on Nutrient agar (Hi Media, Mumbai, India) at 37^oC for 24 h. The culture was sub cultured regularly (every 30 day) and stored at 4^oC. Inoculum preparation - Ten milliliter of distilled water was taken into the screw cap tube and pure colony of fresh culture bacteria was added into the tube and vortex vigorously. The OD (optical density) was measured with the colorimeter and bacterial population were confirmed to be within in 10⁷-10⁸ mL^-1 and then plated out as inoculums. Antimicrobial bioassay - The in vitro antibacterial activities of the test samples were carried out by disc diffusion method. Dried and sterilized filter paper discs (6 mm diameter) were impregnated with known amount of the test substances (extracts) dissolved in solvent (400 μg disc^-1) were placed on nutrient agar medium uniformly seeded with the test bacteria. Standard disc were used as positive and negative control, respectively. These plates were then kept at low temperature (4^oC) for 24 h to allow maximum diffusion of test samples and then incubated at 37^oC for 24 h to allow maximum growth of microorganisms. The test materials having antibacterial activity inhibited the growth of the bacteria and a clear, distinct zone of inhibition was visualized surrounding the disc. The antibacterial activity of the test agents was determined by measuring the diameter of zone of inhibition in millimeter. Determination of minimum inhibitory concentration (MIC) - The minimum inhibitory concentration (MIC) values were determined by broth dilution assay. Petroleum spirit and ethyl acetate extracts were serially diluted to concentrations ranging from 512μg/ml to 2 μg/ml in petroleum spirit and ethyl acetate solvent against three tested pathogenic microorganisms. Finally, the tested samples were incubated at 37°C for 24 hour. The lowest concentration of all extracts that produced no visible bacterial growth was recorded as the MIC.
Statistical analysis - The antibacterial activity was determined by measuring the diameter of zone of inhibition of millimeter scale that was expressed as mean ± SD (Standard deviation of mean). ANOVA single factor was followed by Duncan’s multiple ranges for multiple comparison tests. P< 0.05 was considered statistically significant.

Source of Information: International Journal of Biosciences | IJB |; ISSN: 2220-6655 (Print) 2222-5234 (Online); Vol. 3, No. 7, p. 156-161, 2013

Article Link (Online Journal): http://www.innspub.net; doi: http://dx.doi.org/10.12692/ijb/3.7.156-161

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The results indicated the potential usefulness of Tinospora cordifolia in the treatment of various pathogenic diseases as it may help in the discovery of new chemical classes of antibiotics that could serve as selective agents for the maintenance of human health and may provide biochemical tools for the study of bacterial diseases.

Knowledge Management: Journal