Abstract
This study investigated the possible additive benefit of ejaculate fractionation and colloidal centrifugation on stallion sperm quality. Using an open-end artificial vagina, the sperm-rich fraction (FRAC-1) was separated from the rest of the ejaculate (FRAC-2) and a third sperm sample representing the combined ejaculate was reconstituted post-ejaculation (RAW). Each semen sample was processed for colloidal centrifugation. The percentage of abnormal spermatozoa was 17.8 ± 7.0% in RAW and 14.6 ± 9.5% in FRAC-1 but decreased to 11.4 ± 4.7% and 9.6 ± 6.9% respectively, after colloidal centrifugation. A sperm DNA fragmentation index of 10.9 ± 5.1% was observed in RAW and 7.5 ± 2.4% in FRAC-1 semen collected with the AV but this decreased to 7.8 ± 2.8% and 5.2 ± 2.3% after colloidal centrifugation. The rate of increase in sperm DNA fragmentation during the first 6 h of incubation at 37 ºC was 1.8 ± 0.9% per hour in RAW semen and 2.0 ± 2.0% per hour in FRAC-1 but this significantly decreased to 1.3 ± 1.4% and 0.9 ± 0.8% respectively after colloidal centrifugation. While stallion seminal characteristics can be improved using colloidal centrifugation, further enhancement is possible if the ejaculate is initially fractionated.
Highlights
A sperm DNA fragmentation index of 10.9 ± 5.1% was observed in RAW and 7.5 ± 2.4% in FRAC-1 semen collected with the AV but this decreased to 7.8 ± 2.8% and 5.2 ± 2.3% after colloidal centrifugation
Potential DNA damage associated with reactive oxygen species (ROS) can be prevented by removing damaged or dead spermatozoa and leukocytes by means of colloidal centrifugation, further contributing to maintenance of sperm chromatin integrity (Johannisson et al, 2009; Morrell et al, 2009a,c)
If fractionation of the stallion ejaculate leads to better semen quality, we propose that this technique has the potential to be further enhanced by single colloidal centrifugation (Johannisson et al, 2009; Morrell & Rodríguez-Martínez, 2009; Morrell et al, 2010b)
Summary
Short communication: Stallion sperm DNA stability and ejaculate fractionation terval. A significant improvement was obtained after colloidal centrifugation of RAW semen in terms of progressive motility, abnormal morphology sperm, and sperm DNA stability (as shown by both SDF-T0 and rSDF-T6). We obtained a significantly higher sperm concentration in FRAC-1, which contained sperm samples with significantly higher total motility and lower SDF-T0 (Tables 1 and 2). We compared uncentrifuged FRAC-1 and colloidal centrifuged RAW semen, to explore whether an improvement in seminal characteristics could be achieved using ejaculate fractionation over colloidal centrifugation (Table 2). (a) 175 out of 1200 cells, (b) 250 out of 1200 cells, (c) 214 out of 1200 cells, (d) 115 out of 1200 cells, (e) 137 out of 1200 cells, (f) 270 out of 3600 cells, (g) 771 out of 3600 cells, (h) 393 out of 3600 cells, (i) 186 out of 3600 cells, (j) 279 out of 3600 cells
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