A study was carried out on nine healthy mature breeding bulls (3 each of Gir, Surti and Murrah breed) to evaluate their fresh and frozen semen quality and their interrelationships. The ejaculates immediately after collection were evaluated for routine physico- morphological attributes, including HOS test. The ejaculates (n=72) having >75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and the French mini straws filled were frozen in liquid nitrogen vapour using a programmable biofreezer. Thawing of straws was done at 37°C for 30 sec and assessed for freezability by conventional technique. All the cattle and buffalo bulls donated consistently normal thick creamy yellow and thick milky white semen, respectively. In Gir, Suti and Murrah bulls (n=24 ejaculate each) the seminal attributes such as ejaculate volume (6.69±0.17, 3.12±0.10 and 3.96±0.16 ml, p less than 0.01); initial motility (80.21±0.88, 84.58±0.60 and 84.38±0.76 %, p less than 0.01); total sperm output/ejaculate (9013.85±265.32, 3935.49±259.63 and 5366.48±332.99 million, p less than 0.01) and live sperm (84.71±0.83, 86.17±0.78 and 86.79±0.79 %, p less than 0.05) differed significantly. The mean percentages of post-thaw motile sperm (53.29±1.56, 58.33±1.43 and 59.58±1.20, p less than 0.01); live sperm (59.00±1.95, 67.00±1.59 and 68.42±1.66 %, p less than 0.01); and HOS reactive sperm (48.25±0.78, 44.21±1.29 and 51.54±1.29 %, p less than 0.01) in Gir, Surti and Murrah bulls semen also differed significantly. The variation among the bulls was significant for buffalo breeds in most of their fresh seminal attributes, except HOST, and for post-thaw motility, but not among Gir bulls. The important seminal attributes like motility, live sperm and HOS reactive sperm of fresh and frozen-thawed semen were significantly and positively interrelated in all three breeds of bulls (r = 0.40 to 0.81, p less than 0.05 to 0.01), suggesting that motility and HOST of fresh semen were good predictors of freezability of bovine semen.

A study was undertaken on semen of three mature bulls each of Gir, Surti and Murrah breed to evaluate the comparative motion characteristics and kinematics of their fresh and frozen-thawed spermatozoa by Biovis CASA. The ejaculates (n= 24/breed) having >75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and were assessed. Amongst motility traits, the total motile, rapid progressive motile and slow progressive motile spermatozoa percentage decreased significantly by 23.08 - 30.09, 43.57 - 55.18, 9.12 - 22.75 %, respectively, plessthan0.01), while non-progressive motile sperm (4.78 - 21.48%) and immotile sperm (164.38 - 178.38 %) percentage increased significantly ( plessthan0.01) in frozen-thawed semen compared to that of fresh semen. The post-thaw quality of semen of all three breeds was in acceptable range. The mean values of sperm velocity/kinematic parameters observed in frozen-thawed semen of Gir, Surti and Murrah bulls, based on total motile sperms, viz., average path velocity, curvilinear velocity, straight line velocity, linearity, straightness, beat-cross frequency, amplitude of lateral head displacement, dancing frequency and dancing mean decreased significantly by 13.70 - 17.79; 9.76 - 12.95; 13.28 - 21.90; 7.28 - 9.68; 4.36 - 7.79; 15.56 - 25.15; 8.78 - 10.50; 6.16 - 18.67 and 12.98 - 15.96 %, respectively, as compared to that of their fresh semen samples. However, wobbling index remained almost same for both fresh and frozen semen. All motility traits differed but none of kinematics/velocity traits differed significantly between breeds/species. The values of all velocity parameters for progressive motile sperms were higher than total motile sperms in all three breeds. The effect of freezing-thawing on velocity and kinematic attributes was much less compared to absolute sperm motility, and both the fresh and frozen-thawed sperms behaved identically with respect to their velocity and kinematics. The rapid progressive motile sperm in both fresh (r=0.41 to 0.92) and frozen-thawed (r=044 to 0.88) semen had significant correlations with most of their velocity traits, and the later were significantly and positively or negatively inter-related among each other in semen of all three breeds. It was therefore concluded that cryopreservation process significantly reduces the motility and kinematics attributes of bovine spermatozoa and, CASA analysis of fresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/ kinematics of spermatozoa.

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This investigation was carried out on semen of three healthy mature breeding bulls of Gir breed to evaluate the interrelationships among sperm quality attributes of fresh and frozen-thawed semen assessed by Biovis CASA. The ejaculates (n = 24) having >75% initial motility were diluted @80 million sperm/mL using TFYG extender, filled in French mini straws, and were frozen using a programmable bio freezer after 4 hours of equilibration. The straws were thawed in a water bath at 37°C for 30 sec. The freshly diluted and frozenthawed samples were assessed for routine subjective tests and various motion characteristics/kinematics by Biovis CASA. The Pearson’s correlations for sperm motility and velocity/kinematic parameters of total motile sperm as well as of progressively motile sperm were studied in freshly diluted and frozen-thawed semen. In fresh semen, total motile sperm assessed by CASA had significant (p less than 0.05, 01) correlations with rapid progressive motile sperm (r = 0.46), wobbling index (r = 0.52) and dancing frequency (r = -0.43) in fresh semen. In frozen-thawed semen, it was significantly correlated only with linearity (r = 0.46). The rapid progressive motile sperm in both fresh (r = 0.41 to 0.92) and frozen-thawed (r = 044 to 0.88) semen, however, had significant correlations with most of their velocity traits. Further, the average path velocity (VAP), curvilinear velocity (VCL), straight line velocity (VSL), linearity (LIN), straightness (STR), wobbling (WOB), beat-cross frequency (BCF), amplitude of lateral head displacement (ALH), and dancing mean (DNM) of sperm showed significant positive or negative interrelationships among each other in both fresh (r = 0.41 to 0.91) as well as post-thawed (r = 0.44 to 0.90) semen. Moreover, the correlations of motility and kinematics parameters of total motile sperm in both fresh and frozen-thawed semen were highly significant with velocity/kinematics traits of only progressively motile sperm, and the velocity traits among only motile sperm were highly significantly interrelated in both fresh (r = 0.46 to 0.98) and frozen-thawed (r = 0.43 to 0.93) semen of Girbulls, though the magnitudes of correlations were lower in frozen-thawed semen as compared to fresh semen. Thus, CASA analysis offresh semen for motility and velocity traits could predict the post-thawed sperm motility and velocity/kinematics of bovine semen.

A study was carried out on semen of nine breeding bulls, three each of Gir, Surti and Murrah breeds, to evaluate the comparative motion characteristics and kinematics of their fresh spermatozoa by CASA. The ejaculates (n=72, 24 of each breed) having >75% initial motility were diluted @ 80 million sperm/ml using TFYG extender and were assessed for motion characteristics by CASA. The overall mean values of rapid motile and immotile sperm per cent were observed significantly greater in Gir bulls semen, while total motile and slow motile sperms were apparently higher in buffalo semen. The mean values of sperm velocity/ kinematic parameters observed based on all motile sperms in Gir, Surti and Murrah bulls semen were: average path velocity 50.01±1.25, 48.51±1.03 and 49.14±1.30 μm/s; curvilinear velocity 88.62±1.66, 87.90 ±1.74 amd 88.93±1.69 μm/s, straight line velocity 44.51±1.35, 43.14±1.12 amd 41.73±2.24 μm/s; linearity 50.06±1.42, 48.86±1.32 amd 48.49±1.84 %; straightness 85.17±0.92, 84.97±0.88 and 83.90±1.17 %; wobbling index 57.00±1.17, 55.32±1.05 and 55.30±1.48 %; beat-cross frequency 15.55±0.58, 16.14±0.43 and 14.97±0.54 hz; amplitude of lateral head displacement 2.39±0.18, 2.57±0.12 and 2.31±0.14 μm; dancing frequency 208.34±15.52,225.00 ±10.74 and 211.29±13.03 μm2/s, and dancing mean 5.70±0.46, 6.33±0.35 and 6.33±0.50 μm2/s, respectively. Almost similar trend with little higher values were noted for sperm velocity/kinematics based on only progressively motile sperm, without breed/species variation. The semen of all three breeds behaved identically for sperm kinematics. The bull variation was insignificant for all the traits in all the three breeds.

This study was carried out on semen of nine healthy mature breeding bulls (3 each of Gir, Surti and Murrah breed) to evaluate the motion characteristics and kinematics of their frozenthawed sperms by using computer assisted semen analyzer (CASA). A total of 24 ejaculates from each breed were frozen in Tris fructose yolk glycerol extender using biofreezer. The motion characteristics of frozen-thawed semen assessed by CASA revealed same pattern as of fresh semen. The mean values of total motile vs. immotile sperms in Gir, Surti and Murrah bulls' frozen-thawed semen were 58.