The aim of this study was to isolate and identify yeast species in wine samples from five different countries, and determine the genera of spoilage yeasts. One hundred and thirty four (no. 134) wine samples (86 white and 48 red wine); (30 bottled, 104 tank wine) were used in this study from five countries Slovakia (no. 53), Czech Republic (no. 17), Hungary (no. 6), Spain (no. 32) and Italy (no. 26). Yeasts were cultivated on Malt extract agar supplemented with bromocresol green. After cultivation, yeasts were identified on species level by MALDI-TOF mass spectrometry. From total 285 isolates, we identify 10 genera of yeasts, Saccharomyces (97 isolates, 1 species), Pichia (120 isolates, 5 species), Kregervanrija (6 isolates, 1 species), Zygosaccharomyces (32 isolates, 2 species), Candida (13 isolates, 2 species), Rhodotorula (4 isolates, 1 species), Wickerhamomyces (7 isolates, 1 species) Meyerozyma (4 isolates, 1 species) and genera with only 1 isolale and species: Torulaspora, and Cyberlindnera.