The objective of this research was to determine the species composition of school rooms of various functionalities, the selection of typical species, and the development of assumptions of school environment monitoring. Out of the 151 species identified in the studied school rooms, 52 were grown at a temperature above 25ºC, which constituted 34.4%. Seven species were isolated by incubating the samples at 40ºC, yet it was indispensable only for Acremonium alabamense. Thermophilic fungi constituted 55.3% of the species with the above-indicated Bio-Safety Level status. Out of the identified fungi, one was classified to bio-safety class (Blastomyces dermatitidis), whereas 22 species were claccified as BSL2 (most were: Aspergillus funigatus, A. flavus, A. terreus) and 61 species to class 1. So far, investigations of the indoor bio-aerosol of rooms in this part of Europe have not demonstrated such a wide species spectrum of fungi, as is the case in this study.