Effect of zinc on boar sperm liquid storage.

Storage and transport of liquid boar sperm for artificial insemination (AI) requires the addition of solutions called extenders, which increase the volume of the ejaculate and help preserve its functional characteristics. Yet, the quality of sperm decreases over time primarily due to the increased production of reactive oxygen species (ROS) that damage the plasma membrane. Many commercial extenders are supplemented with additives that mitigate this effect. In semen, zinc is supplied at high concentration on the seminal plasma and helps protect the plasma membrane of sperm. However, zinc in the seminal plasma is diluted and chelated upon addition of extenders for storage, potentially reducing its antioxidant effect. Here we characterize viability, motility, mitochondrial activity, DNA integrity and ROS content of boar sperm diluted with Sus (Medi Nova, Italy) extender supplemented with different concentrations of ZnCl2, at intervals after dilution during 3 days. The ability of sperm supplemented with 2 mM ZnCl2 to fertilize oocytes in vivo of was also tested. Sperm viability was over 82% for all treatments. Mitochondrial integrity analysis, measured by Cytochrome c activity, indicated a protector effect of Zn, noted as a reduced number of sperm with extensive loss of mitochondrial activity. Acrosomal integrity was improved by treatment with all concentrations of ZnCl2 tested. Sperm kinematics were affected by treatment with ZnCl2, showing higher percentage of progressive and rapid sperm in doses supplemented with 2mM ZnCl2. ROS levels and chromatin integrity did not show differences between ZnCl2-supplemented doses and the control. Fertilization rate, total number, live, still born and mummified piglets did not change when sperm were diluted with extender containing 2 mM ZnCl2. The presented characterization indicates that Zn addition to Sus extender have a protective effect on mitochondrial sheath and acrosomal membranes; and provides the basis for further studies aimed to optimize sperm performance in AI.

Heparin binding analysis of boar sperm and its relation with farrowing capacity.

New methods for the evaluation of semen quality according to in vivo reproductive data are useful tools for identifying boars of lower fertility among individuals with standard semen parameters. In this study, indirect fluorescence microscopy was used to evaluate the heparin binding site distribution upon capacitation of sperm from eight boars arbitrarily distributed into Groups I and II according to differential farrowing rates (Group I: ≥ 70%, Group II: < 70%). Additionally, the ability of sperm to bind to solubilised zona pellucida (ZP) was assayed in the presence or absence of heparin. Samples of two individuals of Group II showed lower percentage of B pattern in relation to other individuals (P < 0.001). The number of spermatozoa attached to ZP after 2 h of incubation in capacitating conditions with heparin was significantly lower than in its absence (P < 0.0001). These results suggest that heparin binding site distribution concerning capacitation may be indicative of the availability of proteins involved in the fertilisation process, specifically at the initial sperm-oocyte recognition. Differences in heparin binding site dynamics during capacitation may help identify a subpopulation of individuals with lower fertilising capacity and normal spermiogram, which is particularly useful at high-production establishments.