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Effect of Oxytetracycline on Bacterial Load of Labeo Rohita (rohu) Fish in Culture Pond

Md. Abul Kashem
Faculty of Fisheries, Sylhet Agricultural University

Ferdaus Mohd. Altaf Hossain
Faculty of Veterinary & Animal Science, Sylhet Agricultural University, Bangladesh

Mohammed Masud Hassan
Noakhali Integrated Agro Industries Ltd. Noakhali Sadar, Noakhali, Banglladesh.

Mohammed Nurul Absar Khan
Faculty of Fisheries, Chittagong Veterinary and Animal Sciences University

Syed Ariful Haque
Dept. of Fisheries Technology, Sheikh Fajilatunnesa Mujib Fisheries College

Md. Tawheed Hasan
Faculty of Fisheries, Sylhet Agricultural University

Mohammad Mosarof Hossain
Faculty of Fisheries, Sylhet Agricultural University

Md. Naim Uddin
Dept. of Fisheries Technology, Bangladesh Agricultural University

Abstract/ Executive Summary:

The study was aimed to outline the anti biogram profiling of oxytetracycline against usual ecological bacterial flora of culture ponds of L. rohita. After applying pelleted feeds treated with oxytetracycline (OTC) at 2gm/kg the physicochemical parameters of water like temperature, pH, dissolved oxygen, alkalinity, ammonia and nitrate were recorded weekly: 28.99 to 31.09°C, 7.58-7.95 ppm, 5.36-5.86 mg/L, 86.31-111.99 mg/L, 0.20-0.30 mg/L and 0.11-0.17 mg/L, respectively. The total viable counts of bacteria were found 4.9±1.0³×10³- 5.75±1.0×10³cfu/mL in pond water, 5.62±1.0×10⁷- 6.60±1.02×10⁷cfu/g in sediments, 6.77±1.0×10⁶-7.57±1.0×10⁶ cfu/g in gills, 6.02±1.02×10⁷-8.32±1.0×10⁷cfu/g in gut of L. rohita in control ponds. After OTC treatment the total viable count of bacteria ranged from 3.1±1.19×10³-3.1±1.20×10³cfu/mL in water, 3.1±1.13×10⁶-4.27±1.10×10⁶cfu/g in sediment, 2.82±1.25×10⁵- 3.09±1.19×10⁵cfu/g gill, 2.69±1.12×10⁶- 4.68±1.12×10⁶cfu/g in guts of L. rohita respectively, indicating reduction of overall bacterial load below 1 log in sediment, gills and guts of L. rohita significantly (P<0.005).

Keywords: Bacterial load, Culture ponds, L. rohita, Oxytetracycline

Location: Faculty of Fisheries, BAU, Mymensingh.

Start Date: 00-00-2013

End Date: 00-00-2013

Program Area: Variety and Species

Commodity: Carp fish

Objectives:

1. To determine the bacterial load in water, sediments, gills and intestine of experimental L. rohita before antibiotic treatments and after antibiotic treatments in ponds.

Methodology:

Experimental site: The experiment was conducted in three (3) earthen ponds of the Faculty of Fisheries, BAU, Mymensingh. The ponds were more or less similar in size (12×15 ft), depth, basin conformation, bottom-soil type and contour. Ponds were classified into following groups. a) Control pond without antibiotic medicated diet fed (pond no.-3) b) Oxytetracycline treated ponds (pond no. 1 and pond 2) Pond-1 and 2 were designated as treatment-I and treatment-II where pelleted feed containing 2g/kg ?oxytetracycline was used for two weeks. The ponds were devoid of aquatic vegetations, well exposed to sunlight and completely free from flooding. Rain water and water from deep tube-well were the main sources of water in the experimental ponds during the study period. Physicochemical analysis: Physicochemical parameters were determined for each treatment on weekly basis between 9.00-10.00 hours. Water samples were collected from each pond from surface to a depth of 15-20 cm. On each sampling day, 250 mL water was collected carefully in a clean black plastic bottle with cap from each pond. Each bottle was then marked with respective pond number with three replicates. The physicochemical parameters eg. dissolved oxygen (DO), pH, total alkalinity, ammonia and nitrite-nitrogen and temperature were observed on the spot and were determined by using universal pocket meters. Sampling for bacteriological analysis: For microbial investigations three samplings of water, sediment, gills and intestine of L. rohita were done weekly in culture ponds before antibiotic treatment. After antibiotic treatment samplings were done every alternative day and means were taken. Pond water: 250 ml of water were collected in sterile glass bottles (15- 20 cm depth) from three different locations. The three samples were combined to make a composite sample for bacteriological analysis in the laboratory. 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. Sediment: Bottom sediment 10g were collected, with sterile glass bottles submerged to the bottom, from the same three locations in each pond. The sediment sample of 0.2 g was suspended in 1mL of sterile saline solution. 1 ml of the homogenate was serially diluted (10^-1 to10^-8) and treated as previously described for the water sample. Fish gills and intestine: For every sampling, three L. rohita (80-140 g) from each pond were used for bacterial counts in fish organs (e.g. gills and intestine). The fish were killed by pithing. Around 0.5-1 g each of gills and intestinal content were taken aseptically and homogenized separately in a mortar. Approximately 0.2 g of each homogenate was then putted in a tube containing 2 mL of sterile saline solution. 1 ml of each homogenate solution was serially diluted (to 10^-7) and treated in the same way as the pond water samples. Aerobic plate count (APC): All plates in duplicate on sterile petri dish were done for every dilution. 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 spreaded 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 dark field colony counter (Leica, Buffalo. NY. USA) equipped with a guide plate ruled in square centimetres. Plates containing 30-300 colonies were used to calculate bacterial population results, recorded as cfu per unit of sample. Data analysis: Data were calculated by using simple MS office programme and SPSS software.

Source of Information: Global Advanced Research Journal of Microbiology (ISSN: 2315-5116) Vol. 3(2) pp. 018-024, March, 2014

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The study revealed that, total viable counts of bacteria were found 4.9±1.0³×10³- 5.75±1.0×10³cfu/mL in pond water, 5.62±1.0×10⁷- 6.60±1.02×10⁷cfu/g in sediments, 6.77±1.0×10⁶-7.57±1.0×10⁶ cfu/g in gills, 6.02±1.02×10⁷-8.32±1.0×10⁷cfu/g in gut of L. rohita in control ponds. After OTC treatment the total viable count of bacteria ranged from 3.1±1.19×10³-3.1±1.20×10³cfu/mL in water, 3.1±1.13×10⁶-4.27±1.10×10⁶cfu/g in sediment, 2.82±1.25×10⁵- 3.09±1.19×10⁵cfu/g gill, 2.69±1.12×10⁶- 4.68±1.12×10⁶cfu/g in guts of L. rohita respectively, indicating reduction of overall bacterial load below 1 log in sediment, gills and guts of L. rohita significantly (P<0.005).The numbers of OTC treated ponds bacteria were usually lower in fish ponds undergoing antimicrobial therapy because susceptible microorganisms were inhibited.

Knowledge Management: Journal