Development of sperm vitrification protocols for freshwater fish (Eurasian perch, Perca fluviatilis) and marine fish (European eel, Anguilla anguilla)

Eszter Kása, Gergely Bernáth, Tímea Kollár, Daniel Zarski, Jelena Lujić, Zoran Marinović, Zoltán Bokor, Árpád Hegyi, B. Urbányi, Maria Carmen Vílchez, Marina Morini, David Sanchez Peñaranda, Luz Pérez Igualada, Juan Francisco Asturiano Nemesio, A. Horváth

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19 Citations (Scopus)


Vitrification was successfully applied to the sperm of two fish species, the freshwater Eurasian perch (Perca fluviatilis) and marine European eel (Anguilla anguilla). Sperm was collected, diluted in species-specific non-activating media and cryoprotectants and vitrified by plunging directly into liquid nitrogen without pre-cooling in its vapor. Progressive motility of fresh and vitrified-thawed sperm was evaluated with computer-assisted sperm analysis (CASA). Additional sperm quality parameters such as sperm head morphometry parameters (in case of European eel) and fertilizing capacity (in case of Eurasian perch) were carried out to test the effectiveness of vitrification. The vitrification method for Eurasian perch sperm resulting the highest post-thaw motility (14. ±. 1.6%) was as follows: 1:5 dilution ratio, Tanaka extender, 30% cryoprotectant (15% methanol + 15% propylene-glycol), cooling device: Cryotop, 2. μl droplets, and for European eel sperm: dilution ratio 1:1, with 40% cryoprotectant (20% MeOH and 20% PG), and 10% FBS, cooling device: Cryotop, with 2. μl of sperm suspension. Viable embryos were produced by fertilization with vitrified Eurasian perch sperm (neurulation: 2.54. ±. 1.67%). According to the ASMA analysis, no significant decrease in head area and perimeter of vitrified European eel spermatozoa were found when compared to fresh spermatozoa.

Original languageEnglish
JournalGeneral and Comparative Endocrinology
Publication statusAccepted/In press - Feb 9 2016



  • Eel
  • Fish sperm cryopreservation
  • Perch
  • Spermatozoa vitrification
  • Ultra-rapid cooling

ASJC Scopus subject areas

  • Endocrinology

Cite this

Kása, E., Bernáth, G., Kollár, T., Zarski, D., Lujić, J., Marinović, Z., Bokor, Z., Hegyi, Á., Urbányi, B., Vílchez, M. C., Morini, M., Peñaranda, D. S., Pérez Igualada, L., Asturiano Nemesio, J. F., & Horváth, A. (Accepted/In press). Development of sperm vitrification protocols for freshwater fish (Eurasian perch, Perca fluviatilis) and marine fish (European eel, Anguilla anguilla). General and Comparative Endocrinology.