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Nine healthy mature bulls having average age 3 ± 0.14 years, were selected in three different groups i.e., Sahiwal, Holstein-Friesian and crossbred (Sahiwal x Holstein-Friesian) to compare the semen quality. Semen samples were individually diluted with egg yolk extender and tested for individual motility, progressive motility, plasma membrane integrity, acrosomal and DNA integrity, seminal pH and dead sperm percentage in fresh and thawed semen. Data were subjected to one way analysis of variance. High sperm individual motility (85.12 ± 0.51%), progressive motility (20.25 ± 0.47%), plasma membrane integrity (82.25 ± 0.43%), acrosomal integrity (79.91 ± 0.51%), and DNA integrity (98.54 ± 0.14%) were observed in fresh semen in Sahiwal bulls, which were significantly (P < 0.01) higher compared with the other cattle bulls. High dead sperm percentage (51.87 ± 0.50%), deterioration of plasma membrane integrity (38.29 ± 0.45%), pH (6.83 ±0.01) and DNA integrity (92.45 ± 0.28%) were observed in frozen semen in cross-bred bulls, which showed significant (P < 0.01) differences with the other cattle bulls. The results imply that Sahiwal bulls showed the best semen qualitative characteristics in fresh and frozen semen conditions followed by Holstein Friesian bulls. Moreover, the semen quality declined in the crossbred bull. [ABSTRACT FROM AUTHOR]