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Control of Some Human Pathogenic Bacteria by Seed Extracts of Cumin (cuminum Cyminum L.)

Mominul Islam SHEIKH
Biotechnology and Microbiology Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Soriful ISLAM
Biotechnology and Microbiology Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Atikur RAHMAN
Biotechnology and Microbiology Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Mostafizur RAHMAN
Biotechnology and Microbiology Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh

Mizanur RAHMAN
Department of Biotechnology and Genetic Engineering, Islamic University, Kushtia-7003, Bangladesh

Abdur RAHIM
Department of Genetics and Plant Breeding, Sher-e Bangla Agricultural University, Dhaka-1207, Bangladesh

Abstract/ Executive Summary:

Antibacterial activity of seed extracts of cumin (Cuminum cyminum L.) was investigated against 10 gram positive and gram negative bacteria. Disc diffusion method was used to test antibacterial activity. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined by using standard procedures. The highest (effective) inhibition zone of 16.67±0.47 mm was found at 250 mg/ml for Escherichia coli. On the other hand, the inhibition zones 15.00±0.82 mm for ethanol, 15.33±0.47 for methanol, and 15.67±0.82 for acetone were found against Bacillus subtilis, Sarcina lutea and Klebsiella pneumonia, respectively. MIC value (20 to 50 mg/ml) and MBC value (40 to 60 mg/ml) were measured against studied bacteria. On the basis of investigation, we can say, cumin seeds could be used as a source of new antibacterial agent for developing drugs to inhibit some human pathogenic bacteria.

Keywords: Cumin seed, antibacterial activity, MIC, MBC

Location: Biotechnology and Microbiology Laboratory, Department of Botany, University of Rajshahi, Rajshahi-6205

Start Date: 00-02-2008

End Date: 00-02-2008

Program Area: Crop-Soil-Water Management

Commodity: Cumin

Objectives:

To find out the performance of cumin seeds extracts to control of some human pathogenic bacteria.

Methodology:

Plant material: The Cumin seeds (Cuminum cyminum) were purchased from Municipal market at Kushtia, Bangladesh during February, 2008 and kept in sterilized screw-cap glass container. Samples were crashed and transferred into glass container and preserved it until extraction procedure in the laboratory. Preparation of seed extracts: Cumin seeds were properly dried and pulverized into a coarse powder as reported. Each of 20 g grinded powders was soaked in 60 ml ethanol (95 %), methanol and acetone separately for seven days with occasionally shaking. Aft er seven days of dissolving materials were filtered through Muslin cloth into a beaker and then Whatman no. 1 filter paper were used for final filtration. Th en the filtrates were evaporated using rotary evaporator. At last, 0.97 g of dried extracts were obtained and then stored at 40C in air tight screw-cap tube. Test microorganisms: Ten bacterial species, five gram negative namely Shigella shinga, Shigella dysenteriae, Klebsiellas pneumoniae, Salmonellatyphi, and Escherichia coli and five gram positive namely Sarcina lutea, Bacillus subtilis, Bacillus megaterium, Staphylococcus aureus, and Streptococcus haemolytica were used during this study. All the tested strains were supplied by International Centre for Diarrhoeal Disease Research of Bangladesh (ICDDRB), Dhaka, Bangladesh. Bacteria were grown in Luria-Broth (LB) medium and maintained on nutrient agar slant at 4ºC. Antibacterial assay: To determine the antibacterial activity of seed, cumin seeds with ethanol, methanol and acetone extraction were used. In antibacterial screening, nutrient agar was used as culture media. Th en evaluation of antibacterial activity was performed using standard agar disc diffusion method. Filter paper was cut into small discs using a paper punch and impregnated with 10 μl of different concentration (250, 200, 150, 100 and 50 mg/ml) of seed extracts; dried off and placed on seeded agar plates. 25 μl of bacterial cultures were used for inoculation per agar plates. After inoculation and placing the disc, seeded agar plates were incubated at 37ºC for 24 hours. Aft er incubation, the diameter of zone of inhibition was measured accurately by millimeter scale. Th e results were compared with standard or broad-spectrum antibiotics (Tetracycline at 30 μg/ml) as a positive control. Furthermore, disc was prepared using 95 % ethanol, methanol and acetone instead of cumin seed extracts for negative control. Minimum Inhibitory Concentration and Minimum Bactericidal Concentration: Standard procedure for determination of MIC and MBC of seed extracts of Cuminum cyminum L., were followed as previously described. Five different concentration (60, 50, 40, 30 and 20 mg/ml) of seed extracts were used for MIC determination. 0.5 ml of varying concentration of these extracts were taken into test tubes and 2 ml nutrient broth was added separately; finally a loopful bacterial suspension was introduced in those test tubes. Tubes containing bacterial cultures were incubated at 37^o for 24 hours. Th e lowest concentration of extracts that inhibit growth of selected bacteria in test tubes was taken as MIC. Th e test tubes containing bacterial suspension were considered as a negative control. Series test tubes from the MIC position that did not show any visible growth of bacteria were used for MBC determination. A loopful of broth was collected from tubes and streaked on nutrient agar plates. Aft er streaking the agar plates were incubated at 37^oC for 24 hours. MBC was recorded by using the lowest concentration of extracts at which no visible growth was seen of studied bacteria on agar plates.

Source of Information: Agriculturae Conspectus Scientificus | Vol. 75 No. 1 p 39-44, 2010

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The cumin seed has the important antimicrobial activity against the tested strains. In this regard, the use of spice and their volatile compounds as natural preservatives in food products may be an alternative to the use of chemical additives. The results also indicated that extract of the cumin seed with methanol, has a strong inhibitory activity on some pathogens.

Knowledge Management: Journal