A significant number of stallions produce low quality ejaculates with high sensibility to chilling. Single Layer Centrifugation (SLC) with Androcoll-E TM has been presented as an efficient method of selecting good quality spermatozoa. In the current study, changes in sperm quality (motility, viability, acrosome integrity and DNA damage) occurring during storage at 5˚C for a maximum of 72 h, were investigated. For that, one ejaculate from 12 stallions was split in two aliquots: control and SLC-selected. Both aliquots were chilled and stored at 5˚C and spermatozoa were evaluated for motility, viability and acrosome integrity at 24, 48 and 72 h post collection. DNA damage was evaluated at 48 h post collection using the comet assay. In the SLC-selected aliquots, there was a significant improvement in terms of progressive motility (0 h: P = 0.005; 24 h: P < 0.001; 48 h: P < 0.001; 72 h: P < 0.001) and percentage of live spermatozoa with intact acrosome (24 h: P = 0.003; 48 h: P = 0.003; 72 h: P = 0.004). The DNA damage (in Arbitrary Units) was not different between SLCselected and control samples (P > 0.05). SLC with Androcoll-E TM improved semen quality prolonging sperm longevity of chilled semen (P = 0.012). This positive effect was more evident in ejaculates most sensitive to chilling that had a sharp decrease in motility in the first 24 h of refrigeration and for all ejaculates at 72 h postchilling. Therefore, this method reveals to be a useful technique for selecting spermatozoa and maintain sperm quality during storage.
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