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Determination of Lethal Concentration and Antibacterial Activity of Commonly Used Disinfectants

Alam M
Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh

Rahman MM
Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh

Foysal MJ
Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh

Hossain MN
Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet-3114, Bangladesh

Abstract/ Executive Summary:

The toxic effects of four disinfectants viz., copper sulfate (CuSO₄), potassium permanganate (KMnO₄), methylene blue and malachite green on fish and fish pathogenic bacteria Aeromonas sp., Pseudomonas fluorescens, Edwardsiella sp. and Flavobacterium sp. were investigated. Lethal concentration of the disinfectants to fingerlings of Labeo rohita was determined in aquarium by standard method. Lethal concentration of copper sulfate (CuSO₄), potassium permanganate (KMnO₄), methylene blue and malachite green against fish were found in 0.75ppm, 7ppm, 6ppm and 0.5ppm at 21.4hrs, 18hrs, 9.5hrs and 1.40hrs, respectively. Methylene blue at 4ppm and 5ppm concentration inhibited the growth of Pseudomonas fluorescens and 6ppm concentration suppressed the growth of Aeromonas sp. Copper sulfate (CuSO₄) was effective only against Edwardsiella sp at concentration of 10ppm and 8ppm. Malachite green repressed the growth of all four tasted bacteria at a concentration of 1ppm. Potassium permanganate (KMnO₄) was failed to exhibit any inhibitory effect on the bacteria even at 30ppm concentration.

Keywords: Disinfectant, Lethal concentration, Fish pathogen, Antibacterial activity.

Location:

Start Date:

End Date:

Program Area: Pest Management

Non-commodity: Performance

Objectives:

The present study has been conducted to find out the toxic effects of four disinfectants viz., copper sulfate (CuSO₄), potassium permanganate (KMnO₄), methylene blue and malachite green on fish and fish pathogenic Aeromonas sp., Pseudomonas fluorescens, Edwardsiella sp. and Flavobacterium sp. bacterial isolates.

Methodology:

Determination of Lethal Concentrations to Fish: Determination of lethal concentration of disinfectants copper sulfate (CuSO₄), potassium permanganate (KMnO₄), methylene blue and malachite green were carried out in fish on aquarium, each of 30 liters water capacity. Individual aquarium was suspended with 6 liters of filtered water. Copper sulfate (CuSO₄) solution was prepared at 0.25ppm, 0.5ppm, 0.75ppm, 1ppm, 2ppm, 3ppm, 4ppm, 5ppm, 10ppm, 20ppm and 30ppm concentrations while potassium permanganate (KMnO₄) solution was prepared at 5ppm, 6ppm, 7ppm, 8ppm, 9ppm, 10ppm concentrations. The concentration of methylene blue was 5ppm, 6ppm, 7ppm, 8ppm, 9ppm, 10ppm, 15ppm, 20ppm, 30ppm, 40ppm and 50ppm. Doses of malachite green were 0.1ppm, 0.25ppm, 0.5ppm, 0.75ppm and 1ppm. The concentrations were adjusted by measuring the disinfectants in electric balance and dissolve in necessary volume of water. Each aquarium was marked with specific concentration of individual disinfectant. Six fingerlings of Labeo rohita with a size range of 8-10 cm were stocked in each aquarium and monitored at a regular interval. Determination of Inhibitory Effects on Bacteria: The detrimental activities of the disinfectants against representative of four genera of fish pathogenic bacteria viz., Aeromonas sp. isolate E22, Pseudomonas fluorescens isolate PuKL2, Edwardsiella sp. isolate Eds33and Flavobacterium sp. isolate Fxskin5 were determined in in vitro condition. At the beginning of the experiment the selected isolates were grown in nutrient broth for 24 hours into a shaker-incubator at 25ºC with a rotation of 100 rpm. Approximately 50µl of bacterial broth suspension was taken on nutrient agar plates inside a laminar air-flow cabinet and bacterial broth culture was spared with L-shaped glass rod by turning the plate clock wise and anti-clock wise. The glass rod was sterilized by sinking glass rod with 70% alcohol and flamed in spirit lamp. Solution of different concentrations of copper sulfate (CuSO₄), potassium permanganate (KMnO₄), methylene blue and malachite green were prepared in autoclaved distilled water. Then, 1 ml of each solution was taken in individual micro centrifuge tube and 50 µl of each solution was inoculated on the agar plate containing spread culture inoculums. The plate was remarked with solution concentration and bacterial isolate no. The plates were then kept in an incubator at 25°C for 12 hours. After 12 hours of incubation, the organism was considered sensitive if there was zone of complete inhibition around the inoculated solution. The organism was considered resistant if there was no zone of inhibition.

Source of Information: International Journal of Natural Sciences (2011), 1(4):102-105

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Achievement/Findings:

In the present study, it was found that some disinfectant specially methylene blue and CuSO₄ were active against Aeromonas sp. and Pseudomonas fluorescens at lower than lethal concentration. Findings of the present study will be helpful for fish farmers for prevention and control of bacterial fish diseases.

Knowledge Management: Journal