Our study is one of the very few cases of speleomycological research in recently discovered caves. The aim of this research was to assess the population size of fungal colonies and their species composition in the Jarkowicka cave, discovered in 2012. The air samples were taken from one location outside the cave and from two locations inside of it. Mycological evaluation of the rocks inside the cave was performed usingswab sampling procedure. In the Jarkowicka cave we found 22 species of fungi, including 13 isolated from air at the entrance and from the walls, and 8 species from air inside the cave. Cladosporium spp. were the fungi most frequently isolated from internal atmosphere of the Jarkowicka cave, and from the external air. On the other hand, the fungi most frequently isolated from the rocks were Mucor spp. We found several species not yet described as cave inhabitants: Hypocrea pachybasioides, Cladosporium uredinicola, and Embellisia abundans. Our study may provide a basis for comparison to other similar studies conducted in frequently visited caves by tourists.