Abstract
Although sperm cryopreservation is well developed, different methods can be used for freezing fish semen, mainly with the intention of reducing the use of liquid nitrogen. For this purpose, ultra-freezers could, among other advantages, facilitate the maintenance of semen compared to liquid nitrogen containers. Indeed, ultra-freezers have an increased capacity to store semen, and they can be available in places where there are no other technologies. The aim of this study was to develop an alternative protocol for freezing streaked prochilod Prochilodus lineatus semen using an ultra-freezer. The fish semen (n=10) was collected and frozen using an ultra-freezer (−80°C), with a solution containing 10% DMSO diluted in 5% Beltsville thawing solution in the proportion of 1:4 (semen/diluent) in 0.5-mL straws. Each week, a tube was randomly chosen, thawed, and used to evaluate seminal parameters using computerized semen analysis (CASA). Then, the data were subjected to analysis of variance followed by Tukey’s post hoc test. Motility, motility time, vitality, sperm velocity, and secondary morphology were influenced by storage in ultra-freezer (p < 0.05). However, these parameters were kept at levels considered adequate during the time tested. This is, to the best of our knowledge, the first report of frozen P. lineatus semen under ultra-freezer conditions (−80°C), demonstrating that this method is efficient in maintaining the semen quality for up to 10 weeks.
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Acknowledgements
The authors would like to express their gratitude to Ricardo José da Silva (Estação de Piscicultura da Usina Hidrelétrica da CEMIG) and Ivo Gavião Prado and Fabio Mineo Suzuki (Pisces – Consultoria e Serviços Ambientais) for providing the broodstock and team accommodation as well as Jailson M. Silva (Estação de Piscicultura da Usina Hidrelétrica da CEMIG) for assistance during animal manipulation.
Funding
This work was supported by funds from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (grant number 88882.306651/2018-01), Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (grant number APQ-01070-18), and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (grant number 308359/2019-4).
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Daniella Aparecida de Jesus Paula: Lab work, methodology, investigation, project administration, resources, writing–original draft, writing–review and editing, and supervision experimental design. Naiara Melo: Lab work, writing–original draft, visualization data analysis, and draft editing. William Franco: Data analysis and writing–original draft. Pedro Uzeda: Sampling and sample analysis. Bruna Resende Chaves: Lab work, preparing all equipment, and sampling. Isaac Filipe Moreira Konig: visualization, writing–original draft, and writing–review and editing. Luis David Solis Murgas: Supervision, project administration, visualization, and writing–review and editing.
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de Jesus Paula, D.A., Melo, N., Carneiro, W.F. et al. Cryopreservation of semen in streaked prochilod (Prochilodus lineatus): use of ultra-freezer as alternative technique to liquid nitrogen. Aquacult Int 30, 2629–2639 (2022). https://doi.org/10.1007/s10499-022-00926-3
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DOI: https://doi.org/10.1007/s10499-022-00926-3