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Impact of using a fast‐freezing technique and different thawing protocols on viability and fertility of frozen equine spermatozoa
Author(s) -
Pugliesi G.,
Fürst R.,
Carvalho G. R.
Publication year - 2014
Publication title -
andrologia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.633
H-Index - 59
eISSN - 1439-0272
pISSN - 0303-4569
DOI - 10.1111/and.12205
Subject(s) - semen , membrane integrity , andrology , cryopreservation , insemination , artificial insemination , biology , fertility , sperm , pregnancy , medicine , embryo , membrane , population , biochemistry , genetics , environmental health , microbiology and biotechnology
Summary 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 30 s −1 or 75 °C 7 s −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 20 min 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.