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A stepwise methodology for developing fish sperm cryopreservation protocols: application to the endangered
Jhony Lisbôa Benato1, Nathalia Dos Santos Teixeira2, Rômulo Batista Rodrigues3
1Animal Science Research Program, Federal University of Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brazil.
Cryobiology
|March 14, 2026
Summary
This study developed a successful sperm cryopreservation protocol for the endangered Pogonias courbina, crucial for fish germplasm banking and conservation efforts. The optimized method ensures high post-thaw viability and fertilization rates for this vulnerable species.
Area of Science:
- Aquatic conservation biology
- Fish reproductive physiology
- Cryobiology
Background:
- Wild fish populations face increasing threats from human activities and climate change.
- Aquatic germplasm banking is essential for preserving genetic diversity and supporting aquaculture.
- Developing effective cryopreservation protocols is vital for endangered species conservation.
Purpose of the Study:
- To develop a species-specific sperm cryopreservation protocol for the endangered Pogonias courbina.
- To establish a decision-guided framework for creating fish sperm cryopreservation methods.
- To enable captive breeding, restocking, and germplasm banking for P. courbina.
Main Methods:
- A sequential experimental framework was used to optimize cryopreservation conditions.
- Evaluated various extenders, cryoprotectants (permeable and non-permeable), equilibration times, and thawing parameters.
- Sperm quality assessed via Computer-Assisted Sperm Analysis (CASA); fertilization success verified experimentally.
Main Results:
- Identified optimal thawing at 26°C for 14s and 10 min equilibration.
- The best cryoprotectant combination was HBSS +10% DMSO +5% egg yolk, yielding 71.8% motility (MOT).
- Achieved successful fertilization using cryopreserved sperm, with no significant difference between wild and farmed male sperm.
Conclusions:
- A reproducible Pogonias courbina sperm cryopreservation protocol was established, supporting artificial reproduction.
- The developed framework is generalizable for creating cryopreservation protocols in other fish species.
- This research provides a vital tool for P. courbina conservation and sustainable aquaculture.
