Fresh semen ejaculates as well as frozen thawed semen of a fertile Jersey bull were subjected to certain physio-morphological and in vitro fertilization study in 10 replications. Sperm DNA integrity was assayed with Neutral Comet Assay using single cell gel electrophoresis technique. Mean progressive motility, live sperm and morphologically normal spermatozoa were recorded in fresh and frozen thawed and post capacitated frozen thawed semen of Jersey bull as 78.5 ± 2.11, 65.5 ± 1.89 and 82.5 ± 2.01 per cent; 85.5 ± 1.38, 76.1 ± 1.90 and 88.0 ± 0.82 per cent and 93.4 ± 0.68, 87.3 ± 1.07 and 95.8 ± 0.57 per cent respectively. Mean sperm concentration was found to be 1298.9 ± 69.02 and 83.0 ± 2.41 millions/ml respectively in fresh and frozen thawed semen. The percentage of acrosome reacted spermatozoa was reported to be 2.6 ± 0.34 and 51.8 ± 2.02 per cent respectively in incapacitated frozen thawed and post capacitated frozen thawed semen. Respective comet formation percentage in the present study was reported to be 8.1 ± 0.77, 10.3 ± 0.77 and 4.3 ± 0.30 in fresh, frozen thawed and post capacitated frozen thawed semen. ANOVA revealed that progressive motility, acrosome reaction and comet formation differed significantly (P<0.05) due to different status of semen. Relationship study between different attributes of fresh, frozen thawed and post capacitated frozen thawed semen showed positive and significant (P<0.05) correlation coefficient between motility and morphology in frozen thawed semen, however correlation coefficient of comet formation with motility and morphology in frozen thawed semen was observed to be negative. In vitro maturation and in vitro fertilization percentages were recorded in the present study as 92.31 and 50.81 respectively.