Development of sperm vitrification protocols for freshwater fish (Eurasian perch, Perca fluviatilis) and marine fish (European eel, Anguilla anguilla).
Gen Comp Endocrinol
; 245: 102-107, 2017 05 01.
Article
em En
| MEDLINE
| ID: mdl-27174751
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 15 dilution ratio, Tanaka extender, 30% cryoprotectant (15% methanol+15% propylene-glycol), cooling device Cryotop, 2µl droplets, and for European eel sperm dilution ratio 11, 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.
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Texto completo:
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Preservação do Sêmen
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Motilidade dos Espermatozoides
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Percas
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Criopreservação
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Anguilla
Limite:
Animals
Idioma:
En
Revista:
Gen Comp Endocrinol
Ano de publicação:
2017
Tipo de documento:
Article