Comparative cryopreservation of avian spermatozoa: Effects of freezing and thawing rates on turkey and sandhill crane sperm cryosurvival

Abstract

A comparative approach was used to evaluate semen cooling rates, thawing rates and freezing volume on the cryosurvival of avian sperm. Turkey (Meleagris gallopavo) and sandhill crane (Grus canadensis) sperm were cryopreserved with dimethylacetamide (DMA) concentrations ranging from 6% to 26%. Experiments evaluated the efficacy of (1) rapid, moderate and slow cooling rates, (2) rapid and slow thawing rates, and (3) final volume of semen frozen (0.2 mL compared to 0.5 mL). For crane sperm only, additional experiments were conducted to evaluate the effect of sucrose on cryosurvival. The functionality of frozen/thawed crane sperm was evaluated by fertility trials. For all studies, sperm viability was assessed using the nigrosin-eosin stain. Higher percentages of crane and turkey sperm maintained intact membranes when frozen with moderate or slow cooling rates compared to rapid cooling rates (P<0.05), regardless of DMA concentration. Turkey sperm viability was not affected by thawing rate at any DMA concentration (P>0.05). Crane sperm viability was only affected by thawing rate for the 24% DMA treatment, where moderate thawing was better than slow thawing (P<0.05). Sperm viability was not affected by the semen volume used for freezing for either species (P>0.05). The percentage of membrane-intact crane sperm at lower DMA concentrations was improved by addition of 0.1M sucrose (P<0.05) but not 0.29 M NaCl. The mean fertility rate from frozen/thawed crane semen was 57.5%, and 71.4% of the fertile eggs hatched. The viability of crane sperm was always greater than turkey sperm, regardless of cooling rate, thawing rate or volume of semen frozen. These data verify avian-specific differences in sperm cryosurvival, further emphasize the need for species specific studies to optimize cryopreservation protocols.