title: Development of sperm vitrification protocols for freshwater fish (Eurasian perch, Perca fluviatilis) and marine fish (European eel, Anguilla anguilla).
abstract: 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, 2μl 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 2μl 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.
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary. Electronic address: Kasa.Eszter@mkk.szie.hu.
authors.forename: Eszter
authors.initials: E
authors.lastname: Kása
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Gergely
authors.initials: G
authors.lastname: Bernáth
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Tímea
authors.initials: T
authors.lastname: Kollár
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Daniel
authors.initials: D
authors.lastname: Żarski
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Jelena
authors.initials: J
authors.lastname: Lujić
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary; Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 3, 21000 Novi Sad, Serbia.
authors.forename: Zoran
authors.initials: Z
authors.lastname: Marinović
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Bokor
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Árpád
authors.initials: Á
authors.lastname: Hegyi
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Béla
authors.initials: B
authors.lastname: Urbányi
authors.affiliation: Grupo de Acuicultura y Biodiversidad, Instituto de Ciencia y Tecnología Animal, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: M Carmen
authors.initials: MC
authors.lastname: Vílchez
authors.affiliation: Grupo de Acuicultura y Biodiversidad, Instituto de Ciencia y Tecnología Animal, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: Marina
authors.initials: M
authors.lastname: Morini
authors.affiliation: Grupo de Acuicultura y Biodiversidad, Instituto de Ciencia y Tecnología Animal, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: David S
authors.initials: DS
authors.lastname: Peñaranda
authors.affiliation: Grupo de Acuicultura y Biodiversidad, Instituto de Ciencia y Tecnología Animal, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: Luz
authors.initials: L
authors.lastname: Pérez
authors.affiliation: Grupo de Acuicultura y Biodiversidad, Instituto de Ciencia y Tecnología Animal, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: Juan F
authors.initials: JF
authors.lastname: Asturiano
authors.affiliation: Department of Aquaculture, Szent István University, Páter Károly u. 1., H-2100 Gödöllő, Hungary.
authors.forename: Ákos
authors.initials: Á
authors.lastname: Horváth
mesh_terms: D000811:Anguilla / Q000502:physiology*; D000818:Animals; D015925:Cryopreservation / Q000662:veterinary*; D003451:Cryoprotective Agents / Q000494:pharmacology; D005306:Fertilization; D008297:Male; D000432:Methanol; D010471:Perches / Q000502:physiology*; D055101:Semen Analysis; D012662:Semen Preservation / Q000662:veterinary*; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents; D000432:Methanol
keywords: Eel; Fish sperm cryopreservation; Perch; Spermatozoa vitrification; Ultra-rapid cooling