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

Eszter Kása, Jelena Lujić, Zoran Marinović, Tímea Kollár, Gergely Bernáth, Zoltán Bokor, B. Urbányi, Kinga Katalin Lefler, Dušan Jesenšek, A. Horváth

Research output: Contribution to journalArticle

1 Citation (Scopus)

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 alternative to slow-rate freezing when working in field conditions or when specific equipment necessary for slow-rate freezing is not available.

Original languageEnglish
Pages (from-to)1-9
Number of pages9
JournalFish Physiology and Biochemistry
DOIs
Publication statusAccepted/In press - May 19 2018

Fingerprint

Thymallus
Salmonidae
Vitrification
vitrification
Endangered Species
Trout
Calcium Carbonate
salmonid
marble
sperm
trout
Spermatozoa
spermatozoa
Cooling
Freezing
cooling
motility
cryoprotectants
Propylene Glycol
Equipment and Supplies

Keywords

  • Adriatic grayling
  • Fish sperm cryopreservation
  • Gene preservation
  • Marble trout
  • Spermatozoa vitrification
  • Ultra-rapid cooling

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Aquatic Science

Cite this

Development of sperm vitrification protocols for two endangered salmonid species : the Adriatic grayling, Thymallus thymallus, and the marble trout, Salmo marmoratus. / Kása, Eszter; Lujić, Jelena; Marinović, Zoran; Kollár, Tímea; Bernáth, Gergely; Bokor, Zoltán; Urbányi, B.; Lefler, Kinga Katalin; Jesenšek, Dušan; Horváth, A.

In: Fish Physiology and Biochemistry, 19.05.2018, p. 1-9.

Research output: Contribution to journalArticle

Kása, Eszter ; Lujić, Jelena ; Marinović, Zoran ; Kollár, Tímea ; Bernáth, Gergely ; Bokor, Zoltán ; Urbányi, B. ; Lefler, Kinga Katalin ; Jesenšek, Dušan ; Horváth, A. / Development of sperm vitrification protocols for two endangered salmonid species : the Adriatic grayling, Thymallus thymallus, and the marble trout, Salmo marmoratus. In: Fish Physiology and Biochemistry. 2018 ; pp. 1-9.
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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 alternative to slow-rate freezing when working in field conditions or when specific equipment necessary for slow-rate freezing is not available.",
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AU - Marinović, Zoran

AU - Kollár, Tímea

AU - Bernáth, Gergely

AU - Bokor, Zoltán

AU - Urbányi, B.

AU - Lefler, Kinga Katalin

AU - Jesenšek, Dušan

AU - Horváth, A.

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