Abstract
The diversity of aquatic ecosystems impacted by toxic metals is widely distributed throughout the world. The application of metallothionein (MT) as an early warning sign of metal exposure in freshwater fish is important in biomonitoring, but a more accessible, sensitive, safe, and efficient new methodological strategy is necessary. On this way, a fish MT synthetic gene from Oreochromis aureos was expressed in Escherichia coli to produce polyclonal antibodies against the protein. In the validation assays, these antibodies were able to detect hepatic MT from freshwater fishes Oreochromis niloticus, Pimelodus maculatus, Prochilodus lineatus, and Salminus brasiliensis showing a potential tool for toxic metals biomarker in biomonitoring of aquatic ecosystems. The current results showed the applicability of this molecule in quantitative immunoassays as a sensor for monitoring aquatic environments impacted by toxic metals. Due to the lack of methods focusing on metal pollution diagnostics in aquatic ecosystems, the current proposal revealed a promising tool to applications in biomonitoring programs of water resources, mainly in Brazil where the mining activity is very developed.
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Acknowledgments
The authors would like to thank the Carlos Chagas Institute (ICC-Fiocruz/PR) for allowing MT recombinant expression and purification and the Production and Research Centre of Immunobiological Products (CPPI), Health Department of the Paraná State for housing the rabbits during the immunization period. Thanks also to Extracellular Matrix and Biotechnology of Venoms (UFPR) by Thrombin donation.
Funding
The current study presents part of the results related to the execution of the ANEEL code R&D project PD-06491-0265/2012 carried out by UFPR for COPEL Generation and Transmission SA under the Research and Technological Development Program of the Brazilian Electric sector regulated by the National Agency of Electricity - ANEEL. This research received financial support from Paranaense Energy Company (COPEL) and the National Council for Scientific and Technological Development (CNPq).
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Contributions
PCN: Investigation, protein expression and purification, rabbits immunizations, immunoassay, data curation, and writing ± original draft.
DARV: Polyclonal antibody purification, biological samples preparation, and immunoassay procedure.
MMP: Conceptualization, data analysis, and writing review and editing.
IO: Project supervision and manuscript review.
FCASF: Project supervision and manuscript review.
NITZ: Synthetic gene design, supervision of the protein expression, and purification procedure and manuscript review.
CAOR: Project administration, funding acquisition, general supervision, and writing ± review & editing.
CS: Conceptualization, cleavage reaction, immunoassay supervision, data analysis, figures elaboration, and writing ± review & editing.
Finally, all authors reviewed and approved the final manuscript.
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All animal experiments were approved by the Animal Use Ethics Committee (CEUA) from Biological Sciences Sector at the Federal University of Paraná (CEUA/BIO Number 662, 15/12/2013).
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The authors declare that they have no conflict of the interest.
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Nagamatsu, P.C., Vargas, D.Á.R., Prodocimo, M.M. et al. Synthetic fish metallothionein design as a potential tool for monitoring toxic metals in water. Environ Sci Pollut Res 28, 9517–9528 (2021). https://doi.org/10.1007/s11356-020-11427-2
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DOI: https://doi.org/10.1007/s11356-020-11427-2