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Mastitis, an inflammation of the mammary gland, is one of the most costly and complex diseases of the dairy cows. This study was done to evaluate the occurrence of mastitis pathogens in milk samples from cows with problematic udder health. Samples of milk for bacteriology were taken from dairy cows in an around Nitra region, Slovakia. For this purpose, the samples from udder quarters were cultured and bacteriologically evaluated. From 390 samples 73.85 % of positive samples were found. The predominant bacterial isolates were Coagulase negative staphylococci (17.95 %), followed by Escherichia coli (12.82 %), Staphylococcus aureus (9.74 %), Bacillus spp. (6.41 %), yeasts (5.64 %), Streptococcus uberis (4.1 %), Staphylococcus epidermidis (3.59 %), Pseudomonas aerogenes (3.33 %), others (bacteria and mould) (3.33 %), Entrococcus spp. (3.08 %), Streptococcus agalactiae (1.45 %), Corynebacterium spp. (1.28 %) and Staphylococcus chromogenes (1.03 %). In conclusion, high percentage of positive samples and relatively high occurrence of environmental microorganisms were identified in milk samples indicating the problem with the hygiene of the udder and environment in examined farms.