Maternal heat stress during late gestation affects calf function during postnatal life. The objective of the present study was to evaluate whether calves that experience heat stress in utero have altered thermoregulatory responses to acute heat stress later in life. Specifically, the hypothesis was that heat stress in utero would improve the response to acute heat stress at maturity. Females were born to dams exposed to heat stress or cooled during late gestation preceding their birth. All animals were raised postnatally under identical management. Twelve lactating Holstein cows that were exposed to in utero heat stress (HT) and 12 that were exposed to in utero control (CON) were used. A heat stress challenge was conducted in 3 blocks using 4 HT and 4 CON cows matched according to milk yield, stage of lactation, and parity. Each challenge consisted of transfer from a barn with shade and evaporative cooling to one with shade but no additional cooling for 48 h. The challenge was replicated twice for each block. Sweating rate, respiration rate, rectal temperature (RT), and skin temperature were measured on each cow at 0900, 1100, 1300, 1500, and 1700 h for 2 consecutive days. Mean ambient temperature across 6 challenge days was 26.15 +/- 4.75 degrees C. Tendencies for differences at 1700 h were observed between treatments for RT (HT: 39.5 +/- 0.1; CON: 39.6 +/- 0.1 degrees C; P = 0.065), however, there was no difference in respiration rate (HT: 77.6 +/- 1.6; CON: 79.5 +/- 1.6 bpm; P = 0.85). Sweating rate for shaved skin (HT: 29.4 +/- 2.0; CON: 36.0 +/- 2.0 g/m(2)h; P = 0.057) and for non-shaved skin (HT: 22.5 +/- 1.5; CON: 29.2 +/- 1.2 g/m(2)h; P = 0.01) differed between groups. However, there was no effect on skin temperature at the shaved location (HT: 36.2 +/- 0.2; CON: 36.0 +/- 0.2 degrees C; P = 0.81), but there was a tendency for differences for the non-shaved area (HT: 35.4 +/- 0.2; CON: 34.9 +/- 0.2 degrees C; P = 0.097). Cows that underwent in utero heat stress had greater skin temperature at 1700 h vs. in utero control cows, which may be because HT cows increased skin perfusion, and consequently greater cooling via conduction. In utero HT cows would then have higher heat loss and reduced core body temperature, which results in lower rectal temperature and lower sweating rate when exposed to heat stress. These results support the hypothesis that heat stress in utero in late gestation increases heat tolerance at maturity by increasing capacity to dissipate heat to maintain core body temperature.