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As an attempt to find an inexpensive and simple laboratory method for artificial insemination (AI) bull semen quality assessment, the osmotic resistance of spermatozoa was measured using the hypo- osmotic swelling (HOS) test, developed by Jeyendran et al. (1984) (labelled HOS-1), and its modifications (HOS-2, HOS-3), with decreased osmotic pressure aimed at challenging sperm survival ability. The test results were benchmarked against sperm viability measurements performed using the Computerized Motil- ity Analyzer (CMA), and field fertility was calculated as non-return rate (NRR). Two age groups of Estonian Holstein bull sires were included in this study to test possible age effect on semen quality parameters. The HOS-1 test in fresh bull semen correlated well with sperm general motility (GMot) (r = 0.63, P < 0.001 at batch level and r = 0.77, P < 0.001 at bull level) as well as with progressive motility (PMot) in frozen-thawed (FT) semen (r = 66, P < 0.001 at batch level and r = 0.81, P < 0.001 at bull level), which makes the test suit - able for the prediction of post-thaw semen quality. However, the HOS-2 and HOS-3 values in FT semen had high correlations with NRR ( r = 0.65, r = 0.66, P < 0.001 at batch level and r = 0.63, r = 0.71, P < 0.01 at bull level), which was comparable to those between GMot and NRR or PMot and NRR. A combination of motility parameters and the results of the HOS-1 and HOS-3 tests provided a good model for predicting the potential fertility of bull semen. Values of sperm membrane post-thaw intactness, assessed using HOS-2, as well as of sperm motility measurements were higher in mature bulls compared to those in young bulls. Short conclusion: different modifications of the hypo-osmotic swelling test are useful for routine bovine semen quality assessment at AI stations.

Relationships between the results of hypo-osmotic swelling tests, sperm motility, and fertility in Estonian Holstein dairy bulls