Twenty-seven crossbred calves (1/2 Simmental, 1/4 Hereford, 1/4 Brahman) averaging 257 +/- 11 d of age were either hot-iron-branded (H), freeze-branded (F), or sham-branded (S). Calves were blocked for temperament, weight, and sex and were randomly assigned to day and order in which treatments were applied. To reduce stress from handling at treatment time, each calf was herded through the squeeze chute daily for 5 d before the experiment. Jugular cannulas were inserted in each calf 1 d before application of treatment. Blood samples and heart rate measures were obtained at -5, -3, 0, .5, 1, 3, 5, 10, 15, and 20 min after application of the treatments. Mean concentrations of plasma epinephrine (EPI) were higher for H calves at time .5 min than for either S or F calves (P = .10). To account for individual differences, prebranding heart rates and hormone concentrations were subtracted from subsequent samples and were also used to calculate a proportion for each subsequent sample. Analyses of subtracted values found that EPI concentrations were greater for H calves than for either S or F calves (P = .007) at .5 min postbranding. No other differences were found for the subtracted analyses. Analyses of proportion data also revealed that H calves had greater EPI than did either S or F calves (P = .027) at .5 min postbranding. Only three animals vocalized during branding, one H calf and two F calves. Despite the 5-d acclimation period, handling and restraint elevated plasma cortisol concentrations and heart rate.(ABSTRACT TRUNCATED AT 250 WORDS)