Changes in the milk composition during early, middle and late stage of lactation period were monitored in 10 individual autochthonous donkeys of the Domestic Balkan breed. Animals were grazing ad libitum during early and middle lactation, while supplements (meadow hay, corn and corn stalks) were added during the late lactation period. The aim of this study was to evaluate the main compounds of donkeys? milk during different periods of lactation. Based on the obtained results it was noticed that nutritional effects on milk composition were the greatest in the early and middle lactation. Chemical analysis indicated that donkeys? milk was quite poor in dry matter (8.82-9.68%), fat (0.54-0.67%) and protein (1.44-1.79%), and conversely was high in lactose content (6.06-7.12%). Concentration of vitamin C varied significantly (P<0.05) during the lactation period and was very high (12.84-26.89 ?g/mL) in comparison to milk from other animal species. Likewise, the content of lysozyme (1.95-3.29 g/L) and lactoferrin (1.56-3.14 g/L) during the early and middle lactation period showed an increase. Donkeys' milk did not show any significant differences in MUFA, n6-PUFA and UFA contents, while n3-PUFA, n6/n3 and UFA/SFA ratio showed a significant variability (P<0.05) during the lactation period.