Semen from three fertile stallions was subjected to one of five treatments: 1) freezing in raw form (RAW); 2) lyophilization (LYO); 3) a commercial cryopreser-vation protocol (FREEZE), 4) a commercial cryopreservation protocol, followed by thawing, washing and lyophilization (FREEZE-WASH-LYO), or 5) a commercial cryopreservation protocol, followed by thawing, gradient centrifugation, and lyophilization (FREEZE-GRAD-LYO). Aliquots of semen were subjected to 1 to 3 cycles of freezing or lyophilization. Semen samples were evaluated for chromatin susceptibility to denaturation (%COMP or DFI) using the Sperm Chromatin Structure Assay.
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