The aim of this study was to find out the stress responses of ewes caused by different ways of milking in the terms of dynamics of cortisol and oxytocin concentrations in blood samples. The experiment was performed during two periods of lactation. Forty East Friesian sheep were divided into two groups. The first group (20 ewes) was milked by hand and second group (20 ewes) was milked by machine. Blood samples were taken during evening milking at the beginning and at the end of lactation. The concentration of cortisol was assayed from blood serum and the concentration of oxytocin from blood plasma. Before milking the concentrations of cortisol and oxytocin were lower than after milking (cortisol 0.65 nmol.ml-1 vs. 16.23 nmol.ml-1; oxytocin 4.13 pg.ml-1 vs. 4.98 pg.ml-1). Based on the obtained values it is possible to say that there was not proved significant difference in cortisol and oxytocin concentrations between hand milked sheep and machine milked sheep.