Abstract. The aim of the study was to determine the impacts of different levels of mycotoxin load of Czech dairy herds on the larger scale of the milk indicators including milk physical and technological properties. During three subsequent years individual milk samples (IMSs) were collected from four herds of Czech Fleckvieh (C) and from four herds of Holstein cows (H). The IMSs were collected regularly twice in summer and twice in winter, resulting in a total of 936 IMSs. The feeding rations consisted mainly of conserved roughage and supplemental mixtures according to milk yield and standard demands. Samples of feedstuffs were collected at the same time as IMSs and were analysed for content of deoxynivalenol (DON), fumonisins (FUM), zearalenone (ZEA), aflatoxin (AFL), and T-2 toxin using enzyme-linked immunosorbent assay (ELISA) methods. Based on the mycotoxin load, herds were divided into three groups -Load 1 (negligible, n = 36), Load 2 (low, n = 192), and Load 3 (medium, n = 708). All feedstuff samples were positive for at least one mycotoxin. The most frequently occurring mycotoxins were FUM, DON, and ZEA. Relatively high incidence of AFL (56 % positive samples) was observed. The following milk indicators were influenced by the mycotoxin load of herds: fat, acetone (Ac), log Ac, pH, electric conductivity, alcohol stability, curds quality, curd firmness, whey volume, whey protein, non-protein nitrogen (NPN), urea N in NPN, fat / crude protein ratio, and casein numbers on crude and true protein basis, respectively (P < 0.05). The overall level of mycotoxin load was relatively low, with no clear effect on milk characteristics.