Abstract

Use of sperm separation methods such as density gradient centrifugation for selecting the best spermatozoa for animal breeding is constrained by the problem of dealing with the large volumes of ejaculate produced by the males of some species, such as boars. The purpose of this study was to compare density gradient centrifugation (DGC) with centrifugation on a single layer of colloid (SLC) for the preparation of ejaculated boar spermatozoa using AndrocollTM-P. There was no difference between the two techniques in terms of sperm motility or duration of motility after selection, and sperm motility was retained for at least 24 h longer in the centrifuged sperm preparations than in controls (uncentrifuged aliquots). Sperm motility was significantly better (P<0.001) in the centrifuged sperm preparations (means ± sd: SLC 79.6 ± 8.1% and DGC 74.2 ± 12.0%) than in the uncentrifuged controls (62.9 ± 12.7%). The mean yield of motile spermatozoa for SLC was 67.5 ± 25.6%, and for DGC it was 59.6% ± 22.3% (not significant, ns). Sperm survival was significantly increased by colloid centrifugation (control 3.1 ± 0.3 days, SLC 5.5 ± 0.79 days, DGC 5.75 ± 0.62 days; P<0.001 for uncentrifuged versus centrifuged; SLC vs. DGC, ns). Moreover, boar spermatozoa could be stored for 24 h before centrifugation without any detrimental effect on sperm motility or duration of motility. In a further experiment, larger volumes of ejaculate were processed easily on a modified SLC, indicating that this method may be practical for processing large volumes of boar ejaculates.