Literature data suggest that although there is a great variability of stress responses, animals maintain a pattern of response to different stressful factors. In this context, the objective of this experiment was to verify the different physiological responses of Santa Ines ewes to management conditions considered stressful to the animals. For that, 50 Santa Ines ewes were submitted to 3 different impositions of stress (1: weaning; 2: first milking, 3: change of milker). Cortisol levels of experimental ewes were monitored along lactation and after weaning, 60 d after birth. Plasma samples were collected through jugular puncture in 5 times at −20 min (20 min before the stressful stimulus), 0 min (at the moment of the stressful stimulus) and 60, 120 and 300 min after the stimulus. Hormonal analyses were done using immunoenzymatic kits with readings at 450nm in ELISA equipment at the Laboratory of Animal Physiology of FZEA/USP. Hormonal levels were analyzed in subdivided parcels, which means as repeated measures in time, through MIXED procedure. Means were compared by Tukey test with a significance level of 0.05. For the change of milker stimulus, it were observed greater values in cortisol levels in 0, 60 and 120 min after stress imposition (31.557 ng/mL ± 26.153; 30.106 ng/mL ± 18.138 and 23.989 ng/mL ± 15.982, respectively; P = 0.0069), statistically different from the other times of measurement. In the stress of the first milking, it were observed greater values of cortisol levels 60 min after milking (48.274 ng/mL ± 60.07; P = 0.001) and almost 5 h later, the basal levels were reestablished. Therefore, the pattern of cortisol release in the first milking and weaning differed where in the last, it remained elevated until 300min after stress imposition (40.334 ng/mL ± 59.971), showing that this type of stress can influence the productive life of Santa Ines ewes.