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Oxytetracycline Residues in Thai Pangas Pangasianodon Hypophthalmus Sampled from Sylhet Sadar Upazila, Bangladesh

Md. Motaher Hossain
Department of Fisheries Technology and Quality Control, Sylhet Agricultural University, Sylhet, Bangladesh

A. K. Apurbo Barman
Department of Fisheries Technology and Quality Control, Sylhet Agricultural University, Sylhet, Bangladesh

Md. Matiur Rahim
Institute of Food Science and Technology, Bangladesh Council of Scientific and Industrial Research, Dhaka-1205, Bangladesh.

Mohammad Tariqul Hassan
Institute of Food science and Technology, Bangladesh Council of Scientific and Industrial Research, Dhaka-1205, Bangladesh.

Maksuda Begum
Institute of Food science and Technology, Bangladesh Council of Scientific and Industrial Research, Dhaka-1205, Bangladesh.

Debarshi Bhattacharjee
Department of Fisheries Technology and Quality Control, Sylhet Agricultural University, Sylhet, Bangladesh

Abstract/ Executive Summary:

Intensive culture of Thai Pangas (Pangasianodon hypophthalmus; Sauvage, 1878) in freshwater ponds is much more popular in the aquaculture sector of Bangladesh. Like other cultivated catfishes, pangas is popular among fish farmers and well-known for its faster growth, easy culture system, possessing hardy characteristics, good survival rates, ability to survive at high stocking densities, high disease resistance and tolerance of a wide range of environmental parameters. As a result in existing aquaculture practices of Bangladesh recent extension of commercial pellet-fed culture of pangas and tilapia together now account for near to 29% of total aquaculture production.

Keywords: Freshwater ponds, Pangas, Aquaculture, Oxytetracycline, Hypophthalmus, Fish, Thai Pangas

Location: Markets of Sylhet Sadar Upazila, Sylhet, Bangladesh

Start Date: 00-03-2016

End Date: 00-08-2016

Program Area: Animal Health and Management

Commodity: Pangus

Objectives:

To detect the presence of oxytetracycline residues in Thai Pangas fish.

Methodology:

Experimental fish were collected from four selected fish markets of Sylhet Sadar Upazila, namely Kazir Bazar, Balucharb Noya Bazar, Mejor Tilah Bazar and Tuker Bazar under the study area. For analysis 24 samples of Thai Pangas (850 to 1000 gm) were collected between the periods March 2016 to August 2016, in separate marked polythene bags and were kept in an ice box. Then transported to Microbiology Laboratory of Department of Fisheries Technology and Quality Control, Sylhet Agricultural University and kept in a deep refrigerator. For analyzing purpose then the samples were transported to Food Toxicology Laboratory of Bangladesh Council of Scientific and Industrial Research, Dhaka with icing condition and kept in a deep refrigerator at -20^ºC for promote analysis. Oxytetracycline residue in fish samples was detected by High-Performance Liquid Chromatography (Agilent series 1100). Calibration curve was prepared from injecting corresponding concentrations of oxytetracycline standard solution of 25, 50, 75, 100, 125 and 150 ppb. The detection limit for oxytetracycline was 23.62 ppb. The mean retention times (RT) of the oxytetracyclines were found between 4.031 and 4.25 minutes. After adequate thawing 5.0 g of partially thawed intact samples was taken into 50 ml polypropylene centrifuge tubes. After addition of extraction solution to each sample and homogenization samples were uniformly blended and added to centrifuge tube. Tubes were capped and shaken 10 minutes at speed. Contents of tubes were centrifuged at a minimum 8000 rpm for 20 minutes at approximately 15⁰C. Supernatants were poured into a second centrifuge tube. Then sample were filtered through Whatman No. 1 filter paper. The filtered sample extracts were drained through SPE cartridge attached to an SPE vacuum manifold. A 15 ml graduated centrifuge tube was placed in the vacuum apparatus to serve as a collection vessel and elute oxytetracycline from the cartridge with 6 ml elution solution. Vacuum condition was applied to initiate flow continue elution. Once flow stops, vacuum applied to remove residual solvent from the cartridge. Tubes were removed from vacuum manifold and vortex was done. The tube containing elute was placed in the sample concentrator at the temperature at 40 - 50⁰C to reduce volume of the elute to 0.5 - 0.25 ml under a stream of dry nitrogen. Final volume was adjusted to 1 ml with methanol + water (1 : 1) and briefly vortexing. Then approximately 1.0 ml extract was drawn into a 3 ml syringe and was filtered through a syringe into an HPLC vial (1.5 ml). The remaining extract was stored at -20⁰C. The concentrated extract were subjected to analysis by Agilent 1100 series HPLC system. Mobile phase was prepared by buffer and methanol at a ratio of 70 : 30, injection volume was 20 µl; flow rate: 1 ml/min, column temperature: 30⁰C, detector : fluorescence detector (Agilent 1200 series); excitation wavelength: 380 nm; emission wavelength: 520 nm and run time: 12 minutes. The precision of the method was determined as recoveries of oxytetracycline spiked blank samples. For this two replicate oxytetracycline, free fish samples were spiked with 150 ppb oxytetracycline standard just before test. Of the total 24 Thai Pangas samples obtained from the local fish markets, detectable amount of oxytetracycline residues were detected in 6 (25%) Pangas samples. The range of detected oxytetracycline concentrations of positive samples were between 25.05 and 50.05 ppb (mean = 35.11 ± 3.84 ppb). Whereas, Oxytetracycline residues in 18 (75%) samples of Pangas were found less than the detection limit used for this study.

Source of Information: Bangladesh J. Zool. 46(1): 81-86, 2018 ISSN: 0304-9027 (print) 2408-8455 (online)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

In this study oxytetracycline residues in some sample were found below the detection limit (23.62 ppb). Baydan et al. (2015) reported that no samples of Oblada melanura and Mullus barbatus collected from the natural habitat exceeding the detection limit of oxytetracycline according to their used method. A recent study of Barani and Fallah (2015) reported tetracyclines residues in some trout samples less than the detection limit of competitive enzyme-linked immune sorbent assay method. Mahmoudi et al. (2014) also reported 8.44±6.03 ppb tetracyclines residues in Oncorhynchus mykiss and the confined tetracyclines residues were lower than the detection limit of present study. Turk and Oguz (2016) found no oxytetracycline in fish samples collected from Mu?la province, Turkey. Shim et al. (2010) also reported no oxytetracycline residues in some examined fish during their study. Availability of oxytetracycline residues in contaminated fish in the present study indicates indiscriminate use and avoiding of withdrawal period of this antibiotic. Though detected level of oxytetracycline in examined fish sample did not exceed the maximum residue limit (100 ppb) recommended by European Commission (2010), but in case of long term exposure with higher amount could be hazardous for public health. In this regard some corrective measures are required to ensure safe and antibiotic free fish for mass people. At field level, oxytetracycline use should be kept under supervision and only be used for treatment purposes in fishes with prescribed doses. Fish should be marketed only after achieving the withdrawal period. In addition awareness build up programs and regular residues monitoring of marketed fish by the government authorities are needed. Isolation of antibiotic resistance bacteria in fish and sediments of fish farming areas and antibiotics residues monitoring study of other fish species are also required.

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