Abstract
Research on antioxidant biomarkers can generate profound insights into the defense mechanisms of fish larvae against different stressors and can reveal manipulation strategies for improved growth and survival. However, the number of samples to process and unavailability of required infrastructure in larval-rearing facilities limit the immediate processing, requiring the preservation of specimens. Silver pompano (Trachinotus blochii), a potential marine aquaculture species, shows a low larval survival rate due to poorly developed antioxidant mechanism. In this context, 39 storage conditions, including three storage temperatures and different buffers, were scrutinized to select the most suitable preservation strategy for five important antioxidant biomarkers of fish larvae, viz. catalase activity, superoxide dismutase (SOD) activity, measurement of lipid peroxidation, reduced glutathione (GSH), and ascorbic acid contents. The paper proposes the optimum larval storage conditions for these five evaluated antioxidant biomarkers to generate similar results in preserved and non-preserved larval samples. Larval samples preserved in PBS at lower temperatures (− 20 °C and − 80 °C) are recommended for evaluating catalase activity and ascorbic acid content. Catalase activity can also be evaluated by preserving the larval samples at − 20 °C or − 80 °C without buffers. Larval samples held in PBS or without any buffers at − 20 °C and at − 80 °C were found to be suitable for SOD and GSH evaluation, respectively. Preservation in 50% glacial acetic acid at − 80 °C or − 20 °C was preferred for the lipid peroxidation assays. Apart from methodological perspectives, the paper provides insights into the dynamics of larval antioxidant profiles of T. blochii, for the first time.
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Acknowledgements
The authors are grateful to the the Director, ICAR-CMFRI, Kochi, Dr. Anil M. K., Principal Scientist, Vizhinjam Regional Centre of ICAR-CMFRI and the Head-in-Charge of Vizhinjam Regional Centre of ICAR-CMFRI for providing the research facilities. AJ acknowledges CSIR-UGC for the fellowship and CUSAT (Cochin University of Science and Technology, Cochin) for her PhD registration. This work was supported by the Department of Biotechnology, Government of India funded “Dr. E.G. Silas Centre of Excellence and Innovation (EGS—CoEI) in Marine Fish Microbiome and Nutrigenomics” (BT/AAQ/3/SP28267/2018). GS acknowledges Department of Biotechnology funded EGS-CoEI, and RP acknowledges ICAR funded “Network programme on assessment of antimicrobial resistance (AMR) in microorganisms associated with fisheries and aquaculture in India” for their research fellowship.
Funding
This work was supported by the Department of Biotechnology, Government of India funded “Dr. E.G. Silas Centre of Excellence and Innovation (EGS—CoEI) in Marine Fish Microbiome and Nutrigenomics” (BT/AAQ/3/SP28267/2018).
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AJ performed the experiments, analyzed the results, and drafted the manuscript. STG and SRK conceptualized the presented idea, supervised the findings, helped in the analysis, and provided critical feedback in drafting the manuscript. RP, GS, and DVR provided technical support in conducting and analyzing the results. APG and AMS performed pompano larval production and maintenance. SRK supervised the project and acquired financial support for the project leading to this publication.
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The larval samplings and experiments were done as per the ARRIVE guidelines (Percie du Sert et al. 2020). The live larvae were handled according to the guidelines of U.K. Animals (Scientific Procedures) Act (1986) and EU Directive 2010/63/EU for animal experiments (2019). The procedures for the maintenance and experiments of fish larvae were approved by the ICAR-CMFRI, Kochi, India (BT/AAQ/3/SP28267/2018).
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Jagannivasan, A., Gopakumar, S.T., Sharma S R, K. et al. Profiling the antioxidant biomarkers in marine fish larvae: a comparative assessment of different storage conditions to select the optimal strategy. Fish Physiol Biochem 50, 557–574 (2024). https://doi.org/10.1007/s10695-023-01290-6
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DOI: https://doi.org/10.1007/s10695-023-01290-6