Impact of different dilution techniques on boar sperm quality and sperm distribution of the extended ejaculate

•The addition of semen to extender ('reverse' dilution) does not impair boar sperm quality.•Reverse dilution leads to greater homogeneity of diluted semen.•Reverse dilution allows efficient and safe production of boar semen doses.

The dilution of ejaculates is a fundamental step for the production of liquid-preserved boar semen. For a long time, it has been recommended to add the extender to the ejaculate. The aim of the present study was to first compare the effect of the position ('center' vs. 'wall') where the extender is added to the semen-mixing cylinder (height 32.5 cm; diameter 12.7 cm) using an automatic dispenser (n = 11). In experiment 2 (n = 30), we analyzed the two main dilution methods (extender to the semen ('control') vs. 'reverse'). Experiment 3 was carried out to study the dilution effect on kinematics. In Experiments 1 and 2, the sperm distribution 10 min after the dilution and the sperm quality parameters during long-term storage (d1, d3, d5, and d7) were evaluated. In Experiment 3, sperm quality was assessed during short-term storage at 0, 10, 20, 30 and 60 min after semen dilution ('control' vs. 'reverse'; n = 6). There were no significant differences (P > 0.05) between the treatments in the specific response to bicarbonate, mitochondrial activity, membrane status, thermo-resistance or sperm motility immediately after dilution or long-term storage. The sperm distribution was significantly (P = 0.029) affected by the dilution method in Experiment 2. In summary, treatment with the extender first, which is used by only a few European boar studs, leads to comparable results in sperm quality during storage and better results in sperm distribution after dilution. This procedure is also less time consuming, less foam formation occurs during the semen dilution and the procedure is more hygienic.