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Effects of Processing Procedures on Post-Thaw Acrosomal Retention and Motility of Bovine Spermatozoa Packaged in .3-ml Straws at Room Temperature^{1 ,2,}

The Pennsylvania State University, University Park 16802

Abstract

Ten ejaculates were used to test effects of slow (37.0 min), moderate (7.8 min) or fast (4.2 min) freezing from +5 to –120 C on post-thaw survival of sperm. Semen was extended at 37 C in Tris-yolk-glycerol, packaged in . 3-ml Continental straws at ambient temperature, frozen at one of the three rates and thawed in 95 C water for 7 seconds. For combined post-thaw incubations of 0, 3 and 6 hr at 37 C, acrosomal retention and progressive spermatozoal motility were not significantly different among straws frozen at the three rates.

In a second experiment, 10 ejaculates were used to study effects of cooling time from 25 to 5 C (.5 or 3.5 hr), equilibration time (0, 3 or 9 hr) and thawing method (water at 35 C for 10.0 sec, 65 C for 7.5 sec or 95 C for 6.0 sec). Semen was extended in Tris-yolk-glycerol, packaged in straws at room temperature and frozen at the moderate rate. For combined incubation periods of 0, 3, 6 and 9 hr, acrosomal retention and spermatozoal motility were higher (P<.01) after cooling in 3.5 hr rather than .5 hr if semen was not equilibrated. Also 3 or 9 hr of equilibration was superior (P<.01) to 0 hr of equilibration, and thawing at 65 or 95 C was superior (P<.01) to thawing at 35 C. Interactions of equilibration time with cooling time and thawing method were significant (P<.01). Acrosomal retention and motility of sperm were maximal (P<.01) after 9 hr of equilibration when straws were thawed at 35 C; or after 3 or 9 hr when thawed at 65 or 95 C.

¹ Authorized for publication as Paper No. 5261 in the Journal Series of The Pennsylvania Agricultural Experiment Station on March 8, 1977.

² Partially supported by Atlantic Breeders Cooperative, Lancaster, PA and Northeastern Breeders Association and Sire Power, Inc., Tunkhannock, PA. Continental Plastics Inc., Darien, WI donated supplies. The technical assistance of Mrs. A. Keene, Mrs. H. Homan and Mrs. T. Shirk is gratefully acknowledged.

³ Dairy Breeding Research Center, Department of Dairy and Animal Science.