Spermatozoa from different bucks were stained with different fluorochromes, mixed, and inseminated heterospermically. By altering the interval between insemination and luteinizing hormone injection, spermatozoa were allowed to reside in the female tract approximately 5, 10, or 15 h prior to ovulation. The number of functional spermatozoa, from each male of a pair used, that was transported to the site of fertilization was estimated by counting total number of differently stained spermatozoa that surrounded or fertilized each oocyte. Spermatozoa from split ejaculates within a male competed against each other equally, indicating that the staining procedure did not affect fertilization or functional spermatozoal transport rates. Three pairs of males with high initial semen quality (greater than 80% motility) differed in fertility primarily due to functional spermatozoal transport. Spermatozoal survival in the female tract and capacitation time played a role in differences in male fertility when heterospermic insemination occurred at variable times relative to ovulation. Differences in fertilization not accounted for by spermatozoal transport ratio raised the possibility that rate of egg penetration due to acrosomal enzyme differences may be important in determining male fertility. Therefore, total acrosin, hyaluronidase, and arylsulfatase activity in spermatozoa from specific bucks used in fertilization experiments were determined. Although there were trends favoring high fertility when enzyme content was higher, the difference was significant only for arylsulfatase in one buck.