Optimum thawing temperature and time for bovine semen processed by three methods and packaged in 1 ml ampules

A central composite rotatable design was used to determine optimum temperature and time for thawing semen which was processed in egg yolk-tris-glycerol (ETG), egg yolk-citrate-glycerol (ECG) and skim milk-glycerol (SMG) diluents. Percentage of progressive motility (PPM) and percentage of intact acrosomes (PIA) immediately after thawing (0-hr) and after 3-hr incubation at 37 degrees C were measured. The optimum conditions for temperature and time (C/sec) to sustain maximal PIA response after 3-hr incubation (PIA) were: ETG 42 degrees 121 , ECG 33 degrees 175 , and SMG 38 degrees 128 . Average conditions predicted to be "shared" by all three processing methods were 35 degrees C for 150 sec. Final seminal temperature (FST) was measured at each of these points for ETG, ECG and SMG. Each of the above conditions has a FST above 30 degrees C. A strip plot design was used to compare the optimum and predicted "shared" (35 degrees C 150 sec ) conditions with recommended method (35 degrees C 65 sec ) presently in use, denoted as "present". The predicted "shared" thawing conditions resulted in greater (P<0.01) cell viability (PPM and PIA) than did the "present" thawing method both before and after 3-hr incubation. The viability of semen thawed by the predicted "shared" method did not differ (P<0.05) from that resulting from the optimum thawing condition for each processing method (ETG, ECG and SMG) at 0-hr or 3-hr incubation. Cell viability of skim milk-glycerol processed semen degraded faster over the 3-hr incubation period that did that of ETG or ECG processed semen.