The physiological bases for differences in milk production between breeds of cattle selected for beef or milk production are largely unknown. This study was conducted to determine concentrations of prolactin (PRL) and growth hormone (GH) in serum before and after teat stimulation in primiparous Hereford and Holstein heifers. Blood was collected from 6 beef and 9 dairy heifers at 115, 175, 230 and 250 d of gestation. Sampling times were -15, -10, -5, 0, 2, 4, 6, 8, 10, 12, 15, 20, 25 and 30 min relative to test stimulation. Mean areas under PRL response curves for beef and dairy heifers at 115, 175, 230 and 250 d of gestation were 427, 447, 556, 273 and 243, 189, 167, 343 ng/ml/30 min, respectively. Calculations of area (but not basal levels) excluded instances when no PRL response to test stimulation was obtained (22%). Neither stage of gestation nor breed affected PRL response. Basal PRL did not differ between breeds and was 1.8, 2.6, 2.4 and 9.2 ng/ml at 115, 175, 230 and 250 d of gestation. GH did not differ between breeds and was 6.6, 6.2, 5.5 and 7.4 ng/ml at 115, 175, 230 and 250 d. No difference between breeds was apparent with regard to PRL or GH secretion during first gestation.