Abstract
The aim of this study was to compare the post-thaw distribution of motile sperm subpopulations, following simple or colloid centrifugation. A new analysis was used to evaluate the available number of sperm from each subpopulation after each centrifugation protocol. Frozen/thawed semen samples were divided into the following after-thawing treatments: uncentrifuged control (UDC), sperm washing (SW) and two colloid centrifugation procedures (Equipure, SLC-E, and Androcoll, SLC-A). Percentage of total and progressive motility (TM and PM), as well as sperm motility kinematics, distribution of motile sperm subpopulations, and recovery rates, were statistically compared among treatments. The SLC treatments showed higher (P < 0.001) TM and PM than UDC and SW. Following each SLC procedure, different percentages of the subpopulation with the most vigorous and progressive sperm (sP4) were obtained. SLC-A recovered a larger number of sperm belonging to sP4 than SLC-E, but not significantly higher than SW. From a practical point of view, sperm washing, the standard centrifugation procedure for equine semen processing, recovered the same amount of fast and progressive sperm as colloid centrifugation, apparently the best treatment according to traditional analysis. In conclusion, samples processed by SLC have higher motility percentages than SW and UDC but, after combining the available number of sperm, SLC and SW techniques are equally efficient in recovering sperm from the most vigorous, fast and progressive motile subpopulation (sP4).
Highlights
In sperm selection studies, the distribution of the sperm subpopulations or the percentages of a certain parameter prior to and after centrifugation are compared (Johannisson et al, 2009; Cabrera et al, 2014; Hidalgo et al, 2017; Morrell et al, 2017)
Most of the sperm kinematics were significantly higher in Single layer centrifugation (SLC)-A in comparison to the other post-thaw procedures obtaining values similar to (VCL), or even higher than (VSL, VAP, LIN, STR and WOB) fresh semen (Tab. 1)
This study compared three centrifugation protocols for sperm motility improvement assessed by both a traditional analysis and a new analysis
Summary
The distribution of the sperm subpopulations or the percentages of a certain parameter prior to and after centrifugation are compared (Johannisson et al, 2009; Cabrera et al, 2014; Hidalgo et al, 2017; Morrell et al, 2017). Motile sperm subpopulation analysis works with changes on percentages, and includes only motile sperm (MartinezPastor et al, 2005; Flores et al, 2008; Muiño et al, 2009; Ortega-Ferrusola et al, 2009; Dorado et al, 2010; Dorado et al, 2013a), excluding immotile sperm from the analysis In this sense, a new approach to evaluate sperm subpopulations would allow an understanding of the changes in sperm subpopulations arising from different preparation techniques from a more practical point of view. The aim of this study was, to compare the effect of three centrifugation procedures after thawing on motile sperm subpopulations, assessed by a new analysis that integrates immotile sperm and recovery rate
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