The objective of this study was to evaluate the effects of acclimation to handling on growth, plasma concentrations of progesterone (P4) and cortisol, temperament, and reproductive performance of Brahman-crossbred heifers. Over 2 consecutive years, 37 Braford and 43 Brahman x Angus heifers were initially evaluated, within 30 d after weaning, for BW and puberty status via transrectal ultrasonography and plasma P4 concentrations (d 0 and 10), and for temperament by measurements of chute score, pen score, and exit velocity (d 10 only). On d 11, heifers were stratified by breed, puberty status, temperament score, BW, and age and randomly assigned to receive or not (control) the acclimation treatment. Acclimated heifers were exposed to a handling process 3 times weekly (Mondays, Wednesdays, and Fridays) for 4 wk (d 11 to 39 of the experiment). The acclimation treatment was applied individually to heifers by processing them through a handling facility, whereas control heifers remained undisturbed on pasture. Heifer puberty status, evaluated via plasma P4 concentrations and transrectal ultrasonography, and BW were assessed again on d 40 and 50, d 80 and 90, and d 120 and 130. Blood samples collected before (d 10) and at the end of the acclimation period (d 40) were also analyzed for plasma concentrations of cortisol. Heifer temperament was assessed again on d 40 of the study. No interactions containing the effects of treatment, breed, and year were detected. Acclimated heifers had reduced (P< 0.01) ADG compared with control heifers (0.50 vs. 0.58 kg/d, respectively). Attainment of puberty and pregnancy, however, was hastened (P = 0.02 and 0.04, respectively) in acclimated heifers compared with control. Acclimated heifers had reduced chute score (P < 0.01) and concentrations of cortisol (P < 0.01) and P4 (P = 0.03; prepubertal heifers only) compared with control heifers after the acclimation period (1.37 vs. 1.84 for chute score; 37.8 vs. 50.5 ng/mL of cortisol; 0.52 vs. 0.78 ng/mL of P4). Results from this study indicated that, although acclimation to handling decreased ADG, it resulted in decreased chute score, reduced plasma concentrations of cortisol and prepubertal P4, and enhanced reproductive performance of Brahman-crossbred heifers.