Effects of different equilibration times at 5 °C on boar sperm cryotolerance

Equilibration time (ET) is the period during which sperm cells are in contact with cooling/freezing media components at a temperature of 5 °C, providing a proper osmotic balance between the intra- and extra-cellular milieu. The present study aimed to determine the ET (0, 2, and 4 h) that results in greater post-thaw sperm quality and functions. Based on the post-thaw sperm membrane integrity and motility ratios, 20 ejaculates collected from five boars were classified as having good (GFE, n = 5) or poor (PFE, n = 15) freezing capacity. Ratios of post-thaw sperm with intact plasma membrane and acrosome were similar between ET (0 h: 37.02 % ± 2.85 %; 2 h: 34.59 % ± 7.12 %; 4 h: 37.87 % ± 4.44 %) in GFE samples. In PFE, ratios of sperm with intact plasma membrane and acrosome at post-thaw were greater (P < 0.05) after an ET of 2 h than after an ET of 0 h (2 h: 26.16 % ± 1.54 % and 0 h: 16.74 % ± 1.59 %). Also, ratios of post-thaw sperm with relatively lesser membrane lipids disorder were greater (P < 0.05) after an ET of 2 h than for other ET in both GFE (2 h: 21.97 % ± 4.24 % and 0 h: 16.63 % ± 2.38 %) and PFE (2 h: 16.65 % ± 1.40 % and 0 h: 13.23 % ± 1.25 %) samples. In conclusion, an ET of 2 h results in greater sperm cryotolerance in both GFE and PFE samples, which suggests that modifying the freezing protocol lead to an increase post-thaw sperm function and survival.