The experiment was conducted in a well-known private livestock research and development farm in Bangladesh to find out the variation of semen quality parameter among three generations (F 1 , F 2 and F 3 ) of Holstein Friesian upgraded breeding bulls. A total of 312 ejaculates were collected from 6 upgraded breeding bullsthrought the experimental year. The recorded data were summarized using Microsoft Excel 2010 and analyzed using GraphPad Prism 5 softwere. Out of the 312 ejaculates, 273 (87.50%) were found to be creamy in color followed by 26 (8.33%) and 13 (4.17%) as yellowish and watery, respectively. Generation had significant (P0.05) but initial motility and post freezing motility had the significant differences among the three generations. Initial motility ranges from 75.87±0.32 to 78.40±0.38 percent and the post freezing motility ranges from 50.38±0.41 to 52.16±0.43 percent. It could be concluded that most of the semen quality parameters were influenced by generation and freezing. Semen characteristics were better in F 2 followed by F 3 and F 1 generation in upgraded Holstein Friesian bulls.
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