The objective was to determine factors that affect the expression of estrus. Thirteen lactating Holstein cows were ovariectomized about 4 to 6 wk postpartum and then challenged repeatedly with progesterone and estradiol benzoate to induce estrus six times during the postpartum period. Each challenge included 5 d when the cow was primed with progesterone through insertion of a progesterone-impregnated, foam rubber pessary. Estradiol benzoate (1 mg) was injected intramuscularly 36 h after removal of the pessary. Groups of two to three cows each began the experiment at 3-mo intervals to avoid confounding treated simultaneously. Observations for estrous behavior were at 8-h intervals following each challenge. A minimum of three sexually active cows were always observed together to avoid differences in estrous behavior caused by having too few sexually active animals in the group. Observations for estrous behavior were at 8-h intervals following each challenge. During each observation, cows were observed for 30 min on dirt and for 30 min on concrete. Standing behavior was not influenced by postpartum interval, season of year, or milk yield. Mounting behavior increased from the first to the sixth postpartum challenge, but it was not affected by season of year or milk yield. Duration of estrus, mounting activity, and standing activity were greater on dirt than on concrete. These results indicate that the surface on which cows were observed had a profound effect on sexual behavior; however, postpartum interval, season of year, and milk yield were of minor importance.