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The microbiological characteristics of the grapes are made up of a wide variety of microorganisms, including filamentous fungi. Their presence in grapes is traditionally associated with deterioration in quality. The health of the grapes is very important for obtaining quality wine. The objective of this study was to investigate the diversity of mycobiota on the surface and inside of different grapevine varieties at harvest time in the temperate climate of Slovakia and to identify potentially pathogenic isolates of  Aspergillus  and  Penicillium  producing selected mycotoxins. During the 2021 grape harvest, grapes were collected from the Small Carpathians wine region. Eleven grape samples were analyzed by the plating method and plating method with surface disinfection. Emphasis was placed on  Aspergillus  and  Penicillium  species because of their importance in mycotoxin production. Of the 605 fungal strains detected, 11 genera were identified in the exogenous mycobiota. The most common and abundant genera were  Alternaria  and  Botrytis.  In the genus  Aspergillus ,  A . section  Nigri  is the most abundant, while in the genus  Penicillium ,  P. raistrickii  reached the highest frequency and abundance. Of the 379 strains detected and identified from the endogenous mycobiota, the most common genera were again  Alternaria  and  Botrytis  and the most abundant genus was  Botrytis. Penicillium  species were detected in 17% of all fungi found, with  P. raistrickii  dominating. The  A.  section  Nigri  reached only 4% of the relative density of all isolates. Potentially toxigenic  Aspergillus  and  Penicillium  species were tested for toxinogenity by thin layer chromatography. The most important mycotoxin-producing species found were  A . section  Nigri  but without ochratoxin A production.

Mycobiota in Slovak wine grapes: A case study from the small Carpathians wine region