The objectives of this study were to determine and evaluate effect of breed, age and body condition score (BCS) on qualitative and quantitative parameters of bull’s ejaculate. In total, 16 Holstein (H) and 15 Czech Fleckvieh (F) bulls were collected in September 2009 and May 2010. Volume of semen samples, sperm concentration and percentage of motile spermatozoa were evaluated immediately after collecting. Sperm motility was also evaluated after diluting and freezing/thawing of AI doses and subsequently during the short-term test of sperm survival. Percentages of live and pathologic sperm before diluting also were evaluated. The data set was analyzed using a generalized linear model in SAS/STAT software. A statistically significant effect of the breed, age and body condition on qualitative and quantitative traits of bull’s sperm were determined (P < 0.05–0.01). Sperm activity after collection, dilution and freezing/thawing had significantly decreasing character.