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.

Effect of cryoprotectants and thawing temperatures on chicken sperm quality

“…has been reported to improve the cryosurvival of mammalian, fish and avian spermatozoa (Blanco, Long, Gee, Wildt, & Donoghue, ; He & Woods, ; Mosca et al., ). As described by Blanco, Long, Gee, Wildt, and Donoghue (), the inclusion of sucrose and trehalose in the cryodiluent‐containing DMA as permeating cryoprotectant improved post‐thaw motility of turkey sperm. In a recent study by Mosca et al.…”

Pellet Method of Semen Cryopreservation: Effect of Cryoprotectants, Semen Diluents and Chicken Lines

“…Our results are similar to the report by Mosca et al [17] where sucrose combined with DMA had no influence on post thaw sperm parameters. Species difference in effect of sucrose and trehalose was observed where these osmoprotectants improved post thaw sperm motility in turkey but not in crane [16].…”

“…A combination of penetrating and non penetrating cryoprotectants was tested during semen cryopreservation in rooster and other species [14][15][16][17]. The penetrating cryoprotectants protect cells from cryoinjury by increasing membrane fluidity and reducing the intracellular ice crystal formation [18].…”

Optimization of Sperm Cryopreservation Protocol for Peregrine Falcon (Falco peregrinus)

“…Still, previous studies suggest that the tolerance of avian sperm to changes in cryoprotectant concentrations can be highly species-specific. In several studies comparing different concentrations of DMSO, optimal results were achieved with 4.8% in turkeys [26]; 6% in ring-necked pheasants (Phasianus colchicus), chickens [27], and sandhill cranes [28]; 8% in Indian red jungle fowl (Gallus gallus murghi) [23]; and 10% in ducks (Anas platyrhynchos) [29] and kestrels [30]. Previous studies that examined the effect of extreme osmolarities determined that wild avian species had a higher resistance to hyperosmolar conditions when compared with domestic ones [27,30,31].…”