Four first-calvers of the Black-Pied breed with calves sucking their milk until day 21 of lactation were included in a trial. One suckler cow had four calves. The trial took place within five days which included days 20 and 21 in the first-calvers, that means the last two days with sucking calves (designated as day 1 and day 2 of trial-first and second calf sucking) and days 22 and 24, that means the first and third day after the cows were moved to a cowshed (designated as the third and fifth day of trial-first and third evening machine milking). The first-calvers were separated from the calves and moved to a cowshed at 8 o'clock a.m. on day 3 of trial (Tab. I). The responses to calf sucking were investigated after four-hour prevention of calves to approach the cows (12.00-16.00 p.m.). Blood was sampled with a catheter which was introduced into the vena jugularis a day before the trial outset. Blood samples for cortisol determination were taken in two-hour intervals (when the cows were moved to the cowshed twice in half-an-hour interval and once in an hour interval). Blood samplings were effectuated by day, before, during and after milking, or sucking. Machine milking was finished within 7 to 8 minutes. Two to four calves were intensively sucking a cow within the first 15 minutes, then sucking was irregular. Plasma samples were deep-frozen and stored at -20 degrees C until determinations were performed. Oxytocin and cortisol in the blood plasma were determined by radioassays. In the first evening machine milking (day 3) the second cow yielded 8.23 kg milk while on day 5 in the third milking it was 7.98 kg. The other first-calvers gave by 30-45% less milk on day 3 in comparison with evening milking on day 5 (Tab. II). A trend of the higher average values of oxytocin was observed in machine milking on day 5 of trial if compared with the values recorded during calf sucking, or the first milking (day 3), Fig. 1a, Tab. III. The mean oxytocin concentration during 7-minute milking (12.39 +/- 5.81 pg/ml) was significantly lower in the first-calvers on day 3 in comparison with the mean values recorded on day 5 (28.01 +/- 18.56 pg/ml; P < 0.001), or during sucking (18.32 +/- 7.15 pg/ml; P < 0.01).(ABSTRACT TRUNCATED AT 400 WORDS)