Cattle were of great importance for the Neolithic farmers of southeastern Europe, in particular as farming expanded towards the well-watered regions of Džuljunica (ca. 6200–5500 cal. BCE), one of the earliest known Neolithic settlements in northeastern Bulgaria. The clear stratigraphy and the substantial Bos assemblage from Džuljunica Provided us with a great opportunity to investigate the beginning and evolution of cattle husbandry in the northern Balkans through stable isotope and zooarchaeological analyses. The relative abundance of Bos at Džuljunica leaves no doubt about the importance of beef and cattle herding. Mortality profiles suggest a transition in the early phases of the Neolithic from beef-oriented to mixed beef and milk production husbandry, enabled through intensified post-lactation culling. Stable carbon and oxygen isotope analysis of tooth enamel on a limited number of samples provides no evidence for an extended calving season for increasing milk availability or for vertical mobility. Stable carbon and nitrogen isotope values of bone collagen suggest that cattle were kept near the site, where C3 and C4 plants were available in summer, and that they were occasionally foddered with forest resources in the winter. Cattle experience a diachronic reduction in size on a regional scale, possibly due to farmers' choices aimed at more manageable herds consisting of smaller individuals. Restricting intermixing with local aurochs and the arrival of a new type of cattle may also have contributed to this change. Local factors or inter-regional influences may have influenced the ways cattle husbandry evolved at Džuljunica in particular and in northeastern Bulgaria more generally. More data from the region are necessary to flesh out the role of the interplay among environmental factors, local developments, and inter-regional contacts that facilitate the transfer of skills and traditions relating to the changing modes of cattle husbandry.