Zoophilic Diptera plays a leading role in the epizootic foci formation of many infectious and parasitic diseases and directly affects the quality of livestock products. The current study aimed to analyze the number and species composition of parasitic Diptera in industrial, farm, and homestead agrobiocenoses of large and small cattle, pig, and poultry farms in Eastern Ukraine. The research involved entomological collection during the peak activity daylight hours in early May, July, and early September 2021-2022 per farm. A total of 360 entomological collections were made, and 4310 zoophilous flies were examined. In livestock farms of five districts of the Kharkiv region of Ukraine, 28 species of zoophilic flies were registered, among which Musca domestica, Muscina stabulans, Stomoxys calcitrans, Lucilia sericata, Protophormia terraenovae, and Drosophila species were dominant species. The analysis revealed that cattle biocenoses hosted 27 fly species, pigs had 8 species, and poultry and small cattle each had 7 species. The study indicated an increase in the population of Musca autumnalis, the main species in the pastures, near livestock premises during the summer. Stomoxys calcitrans was also recorded in livestock agrobiocenoses. The species Musca domestica, Musca autumnalis, and Stomoxys calcitrans account for 78.8% to 88.3% of the entire complex of zoophilous flies. The two species of Ortellia caesarion (shiny dung beetle) and Ortellia cornicina (green dung beetle), known for their role as manure mineralizers and deemed non-threatening to animals, were completely absent during the research period. The findings indicated the species of Eristalis tenax in agrobiocenoses in 2021. Therefore, it can be concluded that zoophilic flies are physical irritants to animals and potential carriers of many infectious diseases, especially diseases caused by unicellular organisms.