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Effect of storage and single layer centrifugation before cryopreservation on stallion sperm cryosurvival.
Journal of Equine Veterinary Science 2024 March 18
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 8h/5°C before SC; iv) Storage for 8h/5°C before SLC; v) Storage for 8h/15°C before SC; and vi) Storage for 8h/15°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, 5ºC storage and 15ºC 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 5ºC or 15ºC 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.
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