Development of sperm vitrification protocols for two endangered salmonid species: the Adriatic grayling, Thymallus thymallus, and the marble trout, Salmo marmoratus

Fish Physiology and Biochemistry
Eszter KásaA Horváth

Abstract

Vitrification was applied to the sperm of two endangered fish species of Soča River basin in Slovenia, the Adriatic grayling (Thymallus thymallus) and marble trout (Salmo marmoratus) following testing different cooling devices and vitrifying media. Sperm was collected, diluted in species-specific non-activating media containing cryoprotectants, and vitrified by plunging directly into liquid nitrogen without pre-cooling. Progressive motility, curvilinear velocity, and straightness of fresh and vitrified-warmed sperm were evaluated with computer-assisted sperm analysis (CASA). Fertilization trials were carried out to test the effectiveness of vitrification in the case of grayling. A protocol utilizing a glucose-based extender, 30% cryoprotectants (15% methanol + 15% propylene glycol), 1:1 dilution ratio, and droplets of 2 μl on a Cryotop as cooling device yielded the highest post-thaw motility values for both Adriatic grayling (7.5 ± 6.5%) and marble trout (26.6 ± 15.8%). Viable embryos were produced by fertilizing eggs with vitrified grayling sperm (hatching 13.1 ± 11.7%, control hatching 73.9 ± 10.4%). The vitrification protocol developed in this study can be utilized in the conservation efforts for the two species as an altern...Continue Reading

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Apr 26, 2017·Cryobiology·Jelena LujićÁkos Horváth

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Citations

Feb 18, 2019·Biology of Reproduction·Pierre Comizzoli, William V Holt
Apr 19, 2019·PloS One·Roman FraněkÁkos Horváth
Jan 23, 2021·International Journal of Molecular Sciences·Sylwia JudyckaAndrzej Ciereszko

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Software Mentioned

GraphPad Prism
Cryotop
Sperm VisionTM

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