Production and Evaluation of Frozen Semen of Sahiwal Bull Using Two Different Diluters and Protocols

The experiment was designed to evaluate the effects of diluters and freezing protocols on frozen semen production of Sahiwal breeding bulls. Imported 100% Sahiwal breeding bulls were then reared in the American dairy limited (ADL). Semen samples are processed and frozen once a week in the ADL. Spermatozoa with various motility patterns were collected; these included post-thawing, progressive, fast, slow, circular, and local. Computer Assisted Semen Analyzer (CASA) was used to measure the straight-line velocity (VSL), curvilinear velocity (VCL) and linearity (LIN) parameters. Two diluters (tris egg-yolk and tris egg-yolk aqua) and freezing protocols (slow freezing and fast freezing) were used for frozen semen production of these bulls. The significantly highest (P<0.05) number of post-thawing total, progressive and progressive fast motility (78.42±1.88, 73.50±1.90 and 59.68±2.10%), VCL (115.31±3.82µm/s) and lowest slow, circular and local motility (13.44±1.40, 0.57±0.08 and 4.92±0.41%), LIN (0.39±0.01%) was found in diluter 1 and freezing protocol 2. On the other hand, highest number of circular motility (0.77±0.18%) and VSL (44.83±1.17µm/s) was found in diluter 1 and freezing protocol 1 whereas highest number of slow and local motility (18.84±1.23 and 7.46±0.57%) and LIN (0.61±0.20%) and lowest progressive fast motility (48.73±2.28%), VCL (98.90±3.42µm/s), VSL (39.76±1.33µm/s) was found in diluter 2 freezing protocol 2 and also lowest post-thawing and progressive motility (73.44±1.94 and 66.51±2.02%) was observed in diluter 2 freezing protocol 1. The study reveals that diluter 1(one step egg yolk based) and freezing protocol 1 (5 steps) and 2 (2 steps) seems to be an efficient method for frozen semen production.