A new paper in collaboration with our colleagues from the Angling Club of Tolmin (Slovenia) was accepted for publication in the journal `Fish
Physiology and Biochemistry`.
Kása E., Lujić J., Marinović Z., Kollár T., Bernáth G., Bokor Z., Urbányi B., Lefler K., K., Jesenšek D., Horváth Á. 2018. 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.
Abstract:
Vitrification was successfully 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). Sperm was collected, diluted in species-specific nonactivating media containing cryoprotectants and vitrified by plunging directly into liquid nitrogen without pre-cooling in its vapor. Progressive motility, curvilinear velocity and straightness of fresh and vitrified-warmed sperm were evaluated with computerassisted 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 (hatch after vitrification/warming: 13.1±11.7%, control hatch: 73.9±10.4%). The vitrification protocol developed in this study can be utilized in the conservation efforts for the two species as an alternative to slow-rate freezing when working in field conditions or when specific equipment necessary for slow-rate freezing is not available.
Vitrification was successfully 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). Sperm was collected, diluted in species-specific nonactivating media containing cryoprotectants and vitrified by plunging directly into liquid nitrogen without pre-cooling in its vapor. Progressive motility, curvilinear velocity and straightness of fresh and vitrified-warmed sperm were evaluated with computerassisted 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 (hatch after vitrification/warming: 13.1±11.7%, control hatch: 73.9±10.4%). The vitrification protocol developed in this study can be utilized in the conservation efforts for the two species as an alternative to slow-rate freezing when working in field conditions or when specific equipment necessary for slow-rate freezing is not available.
No comments:
Post a Comment