title: Comparative cryopreservation of avian spermatozoa: benefits of non-permeating osmoprotectants and ATP on turkey and crane sperm cryosurvival.
abstract: A comparative approach was used to evaluate the cryosurvival of turkey and crane sperm frozen in a dimethylacetamide (DMA) cryodiluent supplemented with osmoprotectants and ATP. A range (6-26%) of DMA concentrations was used alone or in combination with ATP (30, 60 or 118mM) or one of the following osmoprotectants: (1) sucrose (turkey, 8.0%; crane, 5.0%); (2) 5.0% sucrose and 5.0% trehalose; or (3) betaine hydrochloride (0.1, 0.2 or 0.4mM). The viability of thawed sperm was assessed using the nigrosin-eosin stain and sperm motility was determined using the hanging-drop technique. For semen frozen only with DMA, post-thaw sperm motility was greatest (P<0.05) for the 6.0%, 10.0% and 18% concentrations, regardless of species. Turkey sperm frozen with the sucrose/trehalose combination had greater (P<0.05) post-thaw motility for all DMA treatments compared to DMA alone. The lowest concentration of the osmoprotectant betaine hydrochloride substantially improved turkey sperm viability post-thaw in all treatments compared to DMA alone (P<0.05). The post-thaw motility of crane sperm was improved (P<0.05) with a combination of 18.0%, 24.0% or 26.0% DMA and 30mM ATP. Moreover, in the presence of osmoprotectants, crane sperm motility decreased as the osmoprotectant concentration increased. The lowest concentration of ATP also improved crane sperm viability post-thaw, especially for DMA concentrations 18% or greater. The combination of sucrose and trehalose improved (P<0.05) crane sperm viability only with 6% and 10% DMA. These data affirm that there are avian-specific differences in sperm survival after cryopreservation and suggest that post-thaw survival can be enhanced by including species-based osmoprotectant/ATP combinations in a cryodiluent where DMA is the cryoprotectant.
authors.affiliation: Aquila Foundation and Center for the Studies on Iberian Raptors, CERI Toledo, Spain.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Blanco
authors.forename: Julie A
authors.initials: JA
authors.lastname: Long
authors.forename: George
authors.initials: G
authors.lastname: Gee
authors.forename: David E
authors.initials: DE
authors.lastname: Wildt
authors.forename: Ann M
authors.initials: AM
authors.lastname: Donoghue
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D001717:Birds / Q000502:physiology; D002463:Cell Membrane Permeability; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods* / Q000662:veterinary; D003451:Cryoprotective Agents / Q000494:pharmacology*; D008297:Male; D055101:Semen Analysis; D012662:Semen Preservation / Q000379:methods* / Q000662:veterinary; D013094:Spermatozoa; D014422:Turkeys / Q000502:physiology; D014882:Water-Electrolyte Balance / Q000187:drug effects / Q000502:physiology
chemical_list: D003451:Cryoprotective Agents; D000255:Adenosine Triphosphate