Abstract
The effects of freezing technique and thawing protocol on thawed semen viability and fertility were studied. Ejaculates from 5 stallions (n=25) were frozen by conventional or a fast-freezing technique. Frozen semen was thawed by two thawing protocols (37°C 30s(-1) or 75°C 7s(-1) ). Thawed semen was evaluated by progressive motility, vigour, morphology and plasma membrane integrity. Mares (n=25) were inseminated with 300 (n=11) or 150 (n=14) million spermatozoa. A greater (P<0.05) vigour and progressively motile spermatozoa were detected, respectively, at thawing and after 20min post-thawing in the fast-freezing technique than in the conventional one. Plasma membrane integrity was also greater (P<0.05) in semen frozen with the fast-freezing technique. Semen viability was not affected by thawing protocol. Pregnancy rate using the fast-freezing technique was 76% (19/25), and did not differ (P>0.05) between insemination doses. We concluded that the 150 million progressively motile spermatozoa per dose using a deep-horn insemination maximises the use of equine semen. The fast-freezing technique, as compared to the conventional one, efficiently preserves the viability and fertilising capacity of spermatozoa, indicating a new method to improve the fertility of frozen equine semen.
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