The study is the first report of the fungal air quality in the Demänovská Cave of Liberty, Slovakia, which is one of the most visited caves in Slovakia (Low Tatras). A total of 108 air samples were collected in June 2014 using the microbiological air sampler "Air Ideal 3P" and Potato Dextrose Agar medium. Fungi were identified based on phenotypic tests and ITS regions analysis. Air samples collected from underground sites contained fewer propagules of fungi (from 86.7 to 126.7 colony-forming units per m 3 of air) than outdoor air samples (391.7). Altogether, the incidence of 18 different fungal species were found in the air of the cave, and most of them were isolated from the indoor samples. Cladosporium macrocarpum spores dominated in this study. The fungal species such as Bjerkandera adusta, Exophiala xenobiotica, Fusarium lateritium, Penicillium aurantiacobrunneum, and Trichoderma citrinoviride were detected for the first time in the internal air of underground sites. Overall, fungal air quality of the Demänovská Cave does not pose a biological threat to people and animals with undamaged immune systems according to most standards of fungal sir contamination. However, some of the airborne fungi detected in the cave can pose a risk to persons and animals with weakened immune systems or people who have fungal allergies.