Egg yolk-Tris-glycerol extender, which is widely used in commercial artificial insemination (AI), was modified by replacing the 20% egg yolk (vol/vol) with a supernatant from egg yolk centrifuged at 50,000 x g for 2 h and then used to cryopreserve bull sperm. Preliminary studies showed that 20% egg yolk interfered with biochemical assays, which could be overcome by centrifugation. In Experiment 1, semen from 4 Holstein bulls was frozen in the experimental Tris extender and compared with the whole milk-glycerol control. A total of 2256 first services resulted in 72.6% 60- to 90-d nonreturns for the control and 71.0% for the Tris extender. In Experiment 2, semen from 10 Holstein bulls was frozen in the experimental Tris extender. Half of the semen was used immediately and half was stored in liquid nitrogen for 1 yr before distribution. The nonreturn rates based on 8878 first services for 7 bulls that completed both parts of the trial were 70.9% initially and 71.6% 1 yr later. This time trend difference of 0.7% was comparable to 0.6% for the AI mean for Holstein sires used at the same time. These fertility results and previous laboratory studies indicate that the conventional egg yolk-Tris-glycerol might be simplified for cryopreserving bull sperm. The experimental Tris extender also was suitable for making biochemical measurements.