The aim of the study was to estimate the influence of the breeding and feeding system on the glycosidase activities in the cow’s milk. Experiment was carried out on three breed milking cow. Cows were divided into two groups: 1st. maintained at winter (TMR. system); 2nd. summer feeding system (pasture and TMR. system). Activities of β. glucuronidase, β. glucosidase, β. galactosidase, hexosaminidase, α. glucosidase and mannosidase in milk were estimated. Activities of all examined glycosidases in whole milk were significantly higher during summer season in milk cow. Differences in β. Gal and α. Glu activity (P≤0.01), in all samples between feeding seasons were observed. Negative correlations between estimated enzymes and milk yield traits (fat, protein and casein) after summer feeding were observed, that may indicate an increase derivative processes in high. yielding dairy cows. Those results show that change of feeding systems in dairy cows leads to increase of enzyme activities in milk which influences on intolerances of lactose in milk.