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Assessing safety, efficacy and residue depletion in golden mahseer, Tor putitora (Hamilton, 1822): biochemical and physiological responses to graded concentrations of oxytetracycline dietary supplementation

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Abstract

The safety and effectiveness of oxytetracycline can potentially manage bacterial infections in fish. This, in turn, might reduce the concerns related to its use in aquaculture and human consumption, such as toxicity, antimicrobial resistance, and other associated risks. The primary objective of this study was to assess how adding oxytetracycline dihydrate to the diet affects its effectiveness, safety, and the presence of residues in T. putitora. T. putitora fingerlings, subjected to experimental infection with Aeromonas hydrophila at a concentration of 108 CFU mL− 1, received an oral administration of oxytetracycline dihydrate. The oxytetracycline dihydrate was added to the feed (corresponding to 2% of the fish body weight) at concentrations of 44.1, 88.2, 132.3 and 176.4 mg Kg− 1 fish body weight per day. This treatment was carried out for 10 consecutive days. The biochemical and physiological responses of T. putitora and efficacy of oxytetracycline dihydrate were determined through estimation of microbial load (CFU mL− 1), haematogram, serum biomarkers, behavioral characteristics, non-specific immunity and residue depletion. Experimentally infected fish showed disease progression and induced histopathological conditions with highest microbial load (CFU mL− 1) in the muscle of both control and treated fish. The fish haematogram showed increased leucocyte and haemoglobin content, influenced by dietary oxytetracycline dihydrate. The fish demonstrated adaptive physiological response to oxytetracycline dihydrate at 44.1 to 88.2 mg and resulted in increased albumin and globulin content. The serum-enzyme assay showed significant increase in aspartate aminotransferase (AST), alanine aminotransferase (ALT) and plasma alkaline phosphatase (ALP) activities in the test fish (< 0.05). Oxytetracycline dihydrate at 88.2 to 132.3 mg Kg− 1 fish body weight per day recorded higher feed intake (75%), significant survivability (66–68%) and histopathological recovery. The suppressed immune response was manifested with decreased respiratory burst and lysozyme activity. The palatability, treatment of bacterial infection, histopathological changes and survivability by fingerlings of golden mahseer determined the safety and optimized the therapeutic potential of the oxytetracycline dihydrate at 88.2 mg Kg− 1 fish body weight per day for 10 days to contain the infection by A. hydrophila. A withdrawal period of 8-d was recommended as oxytetracycline dihydrate concentration depleted below the legal maximum residue limit (MRL 2.0 mg g− 1) in the edible muscle of the golden mahseer reared at an average water temperature of 20 °C. This is considered safe for human consumption.

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Data availability

No datasets were generated or analysed during the current study.

Abbreviations

FDA:

Food and Drug Administration

USFDA:

US Food and Drug Administration

CFU:

Colony forming unit

AST:

Aspartate aminotransferase

ALTL:

Alanine aminotransferase, ALP:Alkaline phosphatase

MMC:

Melano Macrophage Cell

OTC:

Oxytetracycline

CNS:

Central Nervous System

RBC:

Red Blood Corpuscle

MS222:

Tricaine methanesulfonate

MRL:

Maximum Residual Limit

FW:

Freshwater

SW:

Saltwater

BW:

Brackish water

NBT:

Nitro blue tetrazolium

KOH:

Potassium Hydroxide, DMSO:Dimethyl sulfoxide

EDTA:

Ethylenediamine tetraacetic acid

FRP:

Fibre Reinforced Plastic

LD 50 :

Lethal Dose

LC-MS/MS:

Liquid Chromatography Tandem Mass Spectrometry, NBF:Neutral buffered formalin

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Acknowledgements

The authors are thankful to Director, ICAR-Directorate of Coldwater Fisheries Research (ICAR-DCFR), Bhimtal, Nainital, India for the facilities provided under the study.

Funding

This work was funded by Indian Council of Agricultural Research (ICAR), New Delhi, India under the project “All India Network Project on Fish Health (AINP-FH) (Grant F. No. CIBA/AINPFH/2015-16 dated 02.06.2015). The funding was coordinated by ICAR-Central Institute of Brackishwater Aquaculture (ICAR-CIBA), Chennai, India.

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Authors and Affiliations

  1. ICAR-Directorate of Coldwater Fisheries Research (ICAR-DCFR), Anusandhan Bhavan, Bhimtal, Nainital, Uttarakhand, 263 136, India

    Sumanta Kumar Mallik, Neetu Shahi, Krishna Kala, Shivam Singh, Richa Pathak, Ritesh Shantilal Tandel, Amit Pande & Pramod Kumar Pandey

  2. ICAR- Central Institute of Brackishwater Aquaculture (ICAR-CIBA), Annamalai Puram, Chennai, Tamil Nadu, 600028, India

    Prasanna Kumar Patil

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  1. Sumanta Kumar Mallik
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Contributions

S. K. Mallik: conduct the experimental trial and wrote the original manuscript. P. K. Patil: developed the concept and designed the experiment. N. Shahi: analyzed the data. K. Kala, S. Singh and R. Pathak: conducted the experimental trials. R. S. Tandel: analyzed the data. A. Pande: supervised the experimental trial, edited the original manuscript. P. K. Pandey: edited the final manuscript. All authors read and approved the final version of manuscript.

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Correspondence to Sumanta Kumar Mallik.

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Ethics approval and consent to participate

The experimental procedures employed and the utilization of golden mahseer, Tor putitora (Hamilton, 1822) in the investigation were approved by the Institute Animal Care and Use Committee (IACUC) of ICAR-Directorate of Coldwater Fisheries Research, Bhimtal India (File no. ICAR-DCFR/IACUC/07/09/2021/08) and complied with the Institutional Biosafety Committee (IBSC), Department of Biotechnology (DBT) and Ministry of Science and Technology, Government of India in accordance with the Rules for “Manufacture, Use/Import/Export and Storage of Hazardous Microorganisms/ Genetically Engineered Organisms or Cells, 1989 (Rules 1989) of Environment (Protection) Act 1986.” The investigation was conducted following appropriate protocols and guidelines. Furthermore, we confirmed that the techniques and findings presented in the study adhered to the ARRIVE guidelines and regulations.

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Mallik, S.K., Patil, P.K., Shahi, N. et al. Assessing safety, efficacy and residue depletion in golden mahseer, Tor putitora (Hamilton, 1822): biochemical and physiological responses to graded concentrations of oxytetracycline dietary supplementation. Vet Res Commun (2024). https://doi.org/10.1007/s11259-024-10340-0

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