Function of cryopreserved horse semen is improved by optimized thawing rates

The use of cryopreserved semen for artificial insemination in equine breeding programs is increasing because of such benefits as reduction of injuries inherent in natural breeding. In the present study, semen straws from 1 ejaculate from each of 3 stallions predetermined to be poor freezers were thawed after long-term storage in liquid nitrogen (−196°C) at 3 different thaw rates (37°C for 30 seconds, 60°C for 8 seconds, 75°C for 7 seconds). Sperm samples were tested in triplicate for each stallion for motility in caffeine to detect all potentially motile sperm and sperm viability and capacitation status. The fast thaw rate significantly increased the percent of progressively motile sperm compared with the slow (P = .0113) and moderate (P = .0157) thaw rates. The fast thaw rate also improved sperm viability, as measured by the dual stain SYBR-14 and PI (P < .05). Similar improvements were achieved with semen from 3 stallions with average postthaw semen quality. Finally, the chlortetracycline (CTC) stain found that the fast thaw rate reduced the number of sperm undergoing premature acrosome reactions (P = .0329). In summary, thawing frozen equine sperm at 75°C for 7 seconds apparently caused less membrane damage on thawing resulting in enhanced postthaw motility and viability and reduced premature capacitation.