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Antibacterial Activity and Cytotoxicity of Stem Bark of Two Common Plants of Bangladesh

Mahmuda Haque
Department of Pharmacy, Southeast University, Banani, Dhaka, Bangladesh

Md Golam Foysal Chowdhury
Department of Pharmacy, Southeast University, Banani, Dhaka, Bangladesh

Md Faujul Kabir
Department of Pharmacy, Southeast University, Banani, Dhaka, Bangladesh

Md Abdur Rashid
Laboratory of Molecular Signaling, Division of Intramural Clinical and Biological Research (DICBR), National Institute on Alcohol Abuse and Alcoholism (NIAAA), National Institutes of Health (NIH), Bethesda, Maryland, USA

Abstract/ Executive Summary:

Aim: The emergence and spread of antimicrobial resistance is a growing problem in both developing and developed countries and threatens to become a global crisis. In recent years, attempts have been made to investigate indigenous medicines used against infectious diseases, to help in developing safer antimicrobial and anticancer drugs. As part of the further advancement of this research arena, an attempt has been made to study the stem barks of Carica papaya Linn. (C. papaya; family: Caricaceae) and Tamarindus indica Linn. (T. indica; family: Fabaceae), two common plants of Bangladesh. The petroleum-ether, chloroform, and ethyl acetate extracts of the stem bark of both plants were investigated for their antibacterial activity and cytotoxicity. Materials and Methods: The antibacterial activity was evaluated using the disk diffusion method. Cytotoxicity was determined against brine shrimp nauplii. In addition, the minimum inhibitory concentration (MIC) was determined using the serial dilution technique to evaluate antibacterial potency. Results: All crude extracts of T. indica and the chloroform extract of C. papaya appeared very potent in terms of both zones of inhibition and spectrum of activity. However, all the extractives were also subjected to brine shrimp lethality bioassay for preliminary cytotoxicity evaluation. Here, the chloroform extract of C. papaya revealed the strongest cytotoxicity, LC ₅₀ of 10.46 μg/mL. Conclusion: The stem barks of both C. papaya and T. indica show broad-spectrum antibacterial activity and may be potential sources of natural antimicrobial compounds and anticancer agents to be used in the treatment of various infectious diseases caused by resistant microorganisms and of cancer, respectively.

Keywords: Antibacterial activity, Antimicrobial activity, Carica papaya, Cytotoxicity, Tamarindus indica

Location: Rangpur, Bangladesh

Start Date: 00-01-2011

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

We report herein the antibacterial and cytotoxic effects of the stem barks of C. papaya and T. indica for evaluation of their potential use as antimicrobial and cytotoxic agents, to advance research, and to further establish the scientific basis of the traditional uses of these plants.

Methodology:

Collection and identification of the plant

Fresh stem bark samples of the plant C. papaya were collected during the month of January, 2011 from the area of Rangpur, Bangladesh, and fresh stem bark samples of the plant T. indica were collected during the month of July, 2011 from the area of Dhaka, Bangladesh. Both the plants were identified by Dr. Hosne Ara, Director, Bangladesh National Herbarium.

Plant materials extraction and fractionation

The fresh stem barks of both plants were washed, sun-dried, and ground. The powdered stem barks of C. papaya (130 mg) and T. indica (150 mg) were extracted with ethanol at room temperature in flat-bottomed glass containers, with occasional stirring and shaking for 7 days. Next, the extracts were filtered, first through cotton and then through filter paper. The filtrates were concentrated to afford solid masses by using a rotary evaporator. The concentrated ethanol extracts of both plants were made slurry with water, and this was followed by solvent/solvent partitioning with petroleum-ether, chloroform, and ethyl acetate using the Kupchan partitioning method.

Antibacterial test

The antibacterial assay was performed by the disk diffusion technique. The sample solution of the materials to be tested was prepared by dissolving a definite amount of material in the appropriate solvent to attain a concentration of 50 mg/mL. Of this concentrated solution, 10 mL was applied on a sterile disk (5 mm diameter, filter paper) and allowed to dry off the solvent in an aseptic hood. Thus, such disks contain 500 mg of crude extracts. To compare the activity with standard antibiotics, kanamycin (30 mg/disk) was used.

All extracts of the collected plants were tested against five Gram-positive and seven Gram-negative bacteria (Bacillus megaterium, Bacillus subtilis, Bacillus cereus, Staphylococcus aureus, and Sarcina lutea; and Salmonella ^{More Details} paratyphi, Vibrio parahaemolyticus, Vibrio mimicus, Escherichia ^{More Details} coli, Shigella dysenteriae, Pseudomonas aeruginosa, and Shigella boydii). Briefly, in this study the test disks and the standard disk were placed in a Petri dish ^{More Details} seeded with particular bacteria and then left in a refrigerator at 4°C for 12-18 h, in order to diffuse the material from the disks to the surrounding media in the Petri dishes. The Petri dishes were then incubated at 37°C overnight to allow bacterial growth. The antibacterial activities of the extracts were then determined by measuring the respective zone of inhibition in millimeters. The investigation was carried out in triplicate.

Minimum inhibitory concentration (MIC) determination

The MIC is the lowest concentration at which a test sample shows its highest activity against the tested microorganism(s). The MICs of the extracts were also determined by the serial dilution technique against the bacteria. The plant extract (1.0 mg) was dissolved in 2 mL distilled water (two drops Tween 80 were added to facilitate dissolution) to obtain a stock solution. After preparing the suspensions of test organisms (10 ⁷ organisms per microliter), one drop of suspension (0.02 mL) was added to each broth dilution. After 18 h incubation at 37°C, the tubes were examined for bacterial growth. The MIC of the extract was taken as the lowest concentration that showed no growth. Growth was observed in those tubes where the concentration of the extract was below the inhibitory level and the broth medium was observed to be turbid (cloudy). Distilled water with two drops of Tween 80 and kanamycin were used as negative and positive control, respectively. The investigation was carried out in triplicate.

Source of Information: Int J Nutr Pharmacol Neurol Dis 2015;5:75-81

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The results obtained in the present study demonstrated that different extracts (petroleum-ether, chloroform, and ethyl acetate) of the plants C. papaya and T. indica have promising antibacterial properties against the tested microorganisms, which can be utilized in the treatment of infectious diseases caused by resistant microorganisms. The results of the present study also support the traditional usage of the studied plants. Therefore, these results are encouraging enough that the characterization in detail of these fractions in other models can be pursued. Further studies may also be conducted to isolate and purify the active constituents to evaluate their cytotoxicity in human cell line cultures.

Knowledge Management: Journal