Abstract
Abstract The objective of the study was to determine the effect of high hydrostatic pressure (HHP) on quality of cryopreserved semen of young bulls. Semen for this study was collected from 8 bulls aged between 13 and 18 months at monthly intervals, from June to September. After collection, semen was diluted in a commercial Bioxcell® extender (one part at 1:1 and a second part to give a sperm concentration of 20 million/0.2 mL), filled into straws and treated with HHP at 30 MPa for 90 min. After HHP treatment, pre-diluted semen (1:1) was diluted to a sperm concentration 20 million/0.2 mL and filled into straws. In addition, part of the semen diluted to a concentration of 20 million/0.2 mL was not treated with HHP (control). All of it was held at +4°C and frozen in a freezer after 2.5-h equilibration. Semen was thawed in a water bath at 38°C and subjected to estimation of the percentage of motile sperm both subjectively and using a computer-assisted semen analyzer and cytometric assessment of sperm cell membrane integrity. Subjective motility and fast progressive motility were significantly higher with pre-diluted (1:1) and HHP treated semen compared to control (P<0.05). No significant differences were observed in percentage of membraneintact spermatozoa between control and experimental groups. Additionally, the influence of HHP on the sperm of individual bulls was assessed. In bull number 2, the HHP treatment after semen pre-dilution significantly improved progressive motility from 54.1 to 63.4 percent (P<0.05). In bull number 4, the HHP treatment after semen pre-dilution significantly improved subjective motility, rapid motility and progressive motility by 12.5, 16.8 and 16.3 percent, respectively (P<0.05). No effect was seen for 6 bulls. It is concluded that for some bulls, the application of HHP before semen freezing may improve the cryopreservation outcome. However, this requires further research in this area, also to determine the fertilizing capacity of bull semen exposed to high hydrostatic pressure.
Highlights
Bull spermatozoa have a relatively high tolerance for freezing and thawing, and frozen semen has been used in cattle breeding for more than half a century
Research published to date has shown that the limited stress induced by high hydrostatic pressure may increase the cryotolerance, fertilizing ability, and developmental competence of gametes and embryos (Pribenszky et al, 2005, 2007; Du et al, 2008; Pribenszky et al, 2008; Bogliolo et al, 2010)
The fact that the quality parameters of frozen boar semen can improve after high hydrostatic pressure (HHP) treatment was reported by Bryła and Trzcińska (2020)
Summary
Bull spermatozoa have a relatively high tolerance for freezing and thawing, and frozen semen has been used in cattle breeding for more than half a century. One of the technological solutions tested in semen cryopreservation is to use high hydrostatic pressure (HHP) generated in the pressure chamber. In the context of bull semen cryopreservation this solution was first proposed by Pribenszky et al (2007) He reported that HHP (30 MPa for 90 min) significantly improves the postthaw motility of the frozen semen. With proper magnitude and duration, high hydrostatic pressure induces a defensive reaction to stress, increasing resistance to a further stress of cryopreservation. The aim of this study was to determine the effect of high hydrostatic pressure on the quality of cryopreserved semen of young bulls
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