Slow Cooling is Beneficial for Storage of Frozen-Thawed Equine Spermatozoa

Cooled storage of semen after thawing can expand the use of frozen semen, providing the possibility of thawing and evaluating the semen at the storage site and subsequently shipping the semen. Our objectives were (1) to examine the motility and viability of frozen-thawed semen after cooled storage and (2) to compare two cooled-storage protocols for frozen-thawed semen. The samples (n = 31) were either placed immediately in a passive cooling box for 8 or 24 hours (CB) or placed in a refrigerator at 4°C for 30 minutes and then transferred to a passive cooling box (REF). Total and progressive motility were similar at T0 and T8-REF and at T0.5 and T8.5-REF. However, a significant reduction was observed in total motility (-8.12%) between T0 and T8-CB, and in total (-9.96%) and progressive motility (-8.52%) between T0.5 and T8.5-CB (P< .05). A significant reduction was also observed in total and progressive motility between T0 and T24, and between T0.5 and T24.5 for both storage protocols (CB and REF). Viability was lower in T8.5-CB (-11.87%), in T8.5-REF (-9.65%), in T24.5-CB (-13.52%), and in T24.5-REF (-12.32%) compared to T0.5 (P< .05). Our results demonstrate that sperm motility and viability decrease during cooled storage. However, storing the samples at 4°C for 30 minutes before placing the semen in a passive cooling box could mitigate the adverse effect of cooling during short-term storage (8 hours). Additionally, we observed individual variation between samples indicating that this protocol might not be suitable for all stallions. Our data shows that slow cooling and storage of frozen-thawed semen is a valid alternative that allows the expansion of frozen semen in breeding programs.