Post-thaw survival of bovine spermatozoa was compared for semen packaged in either .25- or .5-ml French straws and frozen at three different cooling rates in moving N2 vapor. Using a split ejaculate technique, nine ejaculates were extended in heated skim milk, egg yolk-citrate and egg yolk-Tris and packaged in .25- and .5-ml French straws. Semen packaged in the .25- and .5-ml straws was frozen simultaneously at initial N2 vapor temperatures of -140, -110 or -80 C. Semen was thawed in a water bath at 35 C for 1 min. Recovery of spermatozoa was evaluated immediately post-thaw (0 h) and again after 3 h of incubation at 37 C. Motility estimates and motility counts were made using phase contrast microscopy; percentage of spermatozoa possessing intact acrosomes was quantitated using differential interference contrast microscopy. There was a packaging unit X cooling rate interaction (P less than .05) for all three viability measures. However, there was no consistent trend with regard to cooling rate or packaging unit among the three extenders examined. Post-thaw viability for each characteristic varied (P less than .01) among extenders, but not for cooling rate or packaging unit (P greater than .05). Spermatozoa extended and frozen in egg yolk-Tris had greater (P less than .05) post-thaw viability than those extended in skim milk or egg yolk-citrate regardless of cooling rate or volume of the seminal package.