Effect of storage and single layer centrifugation before cryopreservation on stallion sperm cryosurvival

The objectives of this study were to evaluate the effect of a short, cooled storage before cryopreservation on sperm progressive motility (PM) and compare the effect of different centrifugation methods on post-thaw PM of stored samples. Semen was diluted in chilling extender and aliquoted in 6 protocols: i) Standard centrifugation (SC) followed by freezing; ii) Single Layer Centrifugation (SLC) followed by freezing; iii) Storage for 8 h/5 degrees C before SC; iv) Storage for 8 h/5 degrees C before SLC; v) Storage for 8 h/15 degrees C before SC; and vi) Storage for 8 h/15 degrees C before SLC. PM was assessed before centrifugation, after centrifugation, and post-thawing. Stallions were classified as "good freezers" (GF) or "bad freezers" (BF). The PM in samples immediately frozen was greater than in the stored ones (71.98 +/- 14.2, 52.91 +/- 17.8, 53.93 +/- 18.9 for no storage, 5 degrees C storage and 15 degrees C storage, respectively) (P<0.0001). There was an effect of storage condition (p<0.0001), centrifugation method (p < 0.0001), and freezability (P=0.0016), with an interaction between them (P= 0.0004), on PM after centrifugation. Post-thaw PM was greater in samples treated by SLC than in samples processed by SC, for all storage conditions (p < 0.05). All BF stallions 'showed post-thaw PM < 30 % when samples were previously stored. Storage at 5 degrees C or 15 degrees C for 8 h maintains an appropriate quality in GF stallions. Applying a sperm selection technique as SLC is suggested to improve post-thaw motility, allowing GF straws to be frozen after storage, although BF semen should be prepared by SLC immediately after collection.