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The aim of this study was to evaluate the effect of boar semen storage in different short-term extenders (BTS, Kortowo-3, and M III) on the percentage of spermatozoa showing proapoptotic and necrotic changes. For the first time in this study, Annexin V isolated from swine placenta has been used to determine proapoptotic changes in stored boar spermatozoa. The changes were determined using the IN Cell Analyzer 2000. A gradual decrease in motility was observed on successive days of storage. Spermatozoa incubated in the BTS extender were characterised by the highest average motility, which reached 75% on the 1st d and 39% on day 5. Motility of spermatozoa stored in BTS was significantly higher than those stored in Kortowo-3 and M III extenders after 5 d of storage. Diluted semen contained 1.5% to 2.8% spermatozoa with proapoptotic changes. The discussed process was intensified on the 3rd d of storage when the percentage of apoptotic spermatozoa was determined at 8.3% to 14.6%, and the content of dead spermatozoa exceeded 25%. The analysed extenders differed insignificantly in their ability to protect semen against proapoptotic changes during storage. From the methodological point of view, Hoechst 33258 could be used additionally to stain sperm cells regardless of their status.

Analysis of proapoptotic changes in boar spermatozoa stored at 16 °C in different short-term semen extenders