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Effects of Antibiotic on Bacterial Flora in Mrigal Fish (cirhinus Cirhosus, Bloch, 1795) Under Laboratory Condition

S.A.Haque
Department of Fisheries Technology, Sheikh Fajilatunnesa Mujib Fisheries College, Melandah, Jamalpur, Bangladesh.

M.A.Alam
Department of Fisheries Biology and Genetics, Sheikh Fajilatunnesa Mujib Fisheries College, Melandah, Jamalpur, Bangladesh.

V.C.Roy
Department of Fisheries Technology, Hajee Mohammad Danesh Science and Technology University, Dinajpur.

M.A.Islam
Scientific Officer, Bangladesh Fisheries Research Institute,Shrimp Research Station, Bagerhat, Bangladesh.

M.S.Reza
Department of Fisheries Technology, Faculty of Fisheries, Bangladesh Agricultural University, Mymensingh 2202.

Abstract/ Executive Summary:

The Study was conducted on the effects of antibiotic on bacterial flora under laboratory condition. Oxytetracycline (OTC), the most widely used antibiotic, was fed to the Mrigal (average body weight 25g) at the rate of 2g/kg through fish diet and bacterial content was estimated for a period of 20days.Total 8 aquariums were used, where 3 aquariums for control and 5 aquariums for replication of the treatment. Physico-chemical parameters of aquarium water were determined where temperature,of culture aquariums were more or less similar.Before antibiotic treatment dissolve oxygen (DO), pH and total hardness was 7.80±0.10mg/l, 4.10±0.10, and 890.00±10.00ppm, respectively which reached a value 9.90±0.10mg/l, 5.90±0.10and 710.00±10.00ppm, respectively indicating the changes after 20 days. Prior to antibiotic treatment,bacterial load was 2.90±0.06×10³cfu/ml in aquarium water, 6.90±0.20×10⁵cfu/g in fish gills, 4.70±0.10×10⁷cfu/g in fish intestine,and 85.25±3.38×10⁵in fish skin respectively which was Significantly reduced to 1.25±0.03×10³cfu/ml in water, 5.42±0.20×10⁵cfu/g in gills, 3.33±0.05×10⁷ cfu/g in intestine, and 11.24±0.01×10⁵in fish skin respectively after 20 days treatment period. Water and fish samples were also analyzed for bacteria were completely absent before and after antibiotic treatments.

Keywords: Antibiotic, Physico-chemical parameters, Bacterial load, Cirhinus cirhosus

Location: Laboratory of Fish Harvesting, Dept. of Fisheries Technology, BAU. Implemented by Sheikh Fajilatunnesa Mujib Fisheries College

Start Date:

End Date:

Program Area: Animal Health and Management

Commodity: Carp fish

Objectives:

To determine the changes in its physico-chemical parameters and bacterial population.

Methodology:

Experimental site: Eight glass aquariums (size 37cm×30cm×60cm) were set at the Laboratory of Fish Harvesting, Dept. of Fisheries Technology, BAU , where 3 aquariums for control treatment, and 5 aquariums for antibiotic treatment.These aquaria were filled to a depth of 15 cm with tape water collected from the Dept. of Fisheries Technology laboratory, BAU. The experimental aquariums were designed as control aquarium-1, control aquarium-2, control aquarium-3, and aquarium-1, aquarium-2, aquarium-3, aquarium-4, aquarium-5, for the convenience of study. The mrigal fry was collected from Field Laboratory Complex of Faculty of Fisheries, BAU and released the aquarium. In the aquarium-1,aquarium-2,aquarium-3, aquarium-4 and aquarium-5 feed contained 2g/kg oxytetracycline and was used for 3 weeks. The aerators were set with each aquarium. The aquarium water was changed daily. Physico-chemical parameters were determined for each aquarium 4 days interval. Physicochemical analysis: The water quality parameters were recorded 4 days interval throughout the experimental period. Water quality measurements and sample collection were made between 9.00am to 11.00 am on each sampling Day. Water samples were collected from each aquarium to a depth of 35 cm. On each sampling Day, 500 ml water was collected in a clean black plastic bottle with cap from each a very carefully without any agitation. Each bottle was then marked with respective aquarium number with three replicates. Water quality parameters such as temperature, dissolved oxygen, hardness, and pH were measured. Water temperature was recorded in the laboratory with the help of a centigrade thermometer (div=0.1°C). Other Parameters such as dissolved oxygen, hardness and pH were measured using HANNA Test kit, Hanna Instruments Ltd., Germany. Microbiological analysis: Sampling was done every alternative day for microbial analysis of water, fish gills,intestine, and skin for antibiotic treated fish.Aquarium water samples were collected in sterile glass bottles (250 ml) 15-20 cm below the water surface from three different locations in each aquarium in every sampling. Three samples were combined to make a composite sample for bacteriological analysis in the laboratory. Appropriate sample dilutions were made (10^-1 to 10^-5) with sterile physiological saline (0.85% w/v NaCl) in deionized water. Aliquots of 0.1 ml of the serial dilutions were inoculated onto nutrient media in duplicate using the spread plate method as this medium recovered most of the bacteria. All plates in duplicate on sterile petridish were done on sterile nutrient agar media. From sample solution of different dilutions 0.1 ml samples were taken by a micropipette and transferred aseptically into the pre-prepared agar plates by raising the upper lid sufficient enough to enter the tips of the pipette. The samples were then spread homogenously and carefully by sterile flamed L-shaped glass rod throughout the surface of the media until the sample were dried out. For total heterotrophic aerobic bacterial counts of pond water, sediment, gills and intestine, all the inoculated plates were incubated at 28?C for 24-48 hrs. The colony-forming units (cfu) were counted under a Quebec darkfield colony counter (Leica, Buffalo. NY. USA) equipped with a guide plate ruled in square centimeters. Plates containing 30-300 colonies were used to calculate bacterial population results, recorded as cfu per unit of sample. Determination of Escherichia coli and Salmonella spp. by selective media: Escherichia coli and Salmonella spp. were determined by using selective media EMB-agar and SSagar, respectively. Bacterial inoculums from the spread plate were transferred into the selective media. Growth of bacterial colony in EMB-agar and SS-agar media indicate the presence of E. coli.and Salmonella spp., concurrently. Statistical Analysis: The data obtained in the experiment were recorded and preserved in computer. The data obtained in the experiment were analyzed by using CRD of MSTAT programmed. Significant differences were determined among treatments at the 5 % level (p< 0.05).

Source of Information: European Journal of Food Science and Technology Vol.2, No.1, pp.1-7, June 2014

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Thephysicochemical and bacteriological analysis of the aquarium fishes before treatment and after treatment were done carefully. The Oxytetracycline was used for aquarium treatment. Oxytetracycline is widely used as a antibacterial agent. Initial load of bacteria has great contribution to the quality of end product. Before antibiotic treatment whole physicochemical and bacteriological study of experimental fishes were done. Antibiotic treatment was done at 2g/kg through pelleted feed. After antibiotic treatment both physicochemical and bacteriological analysis were done. In the physicochemical syudy after antibiotic treatment the pH and dissolve oxygen increased gradually,but the total hardness reduced destrically. Bacterial loads were reduced significantly in aquariums when antibiotic medicated diets were used. Escherichia coli and Salmonella spp. were not detected throughout the experiment period.

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