Bio-aerosols transported by the air have been considered as the major source of dispersal agents that contaminate agricultural products. Unseen fungal spores are known sources of bio-aerosols that harm mushroom and human health during mushroom cultivation. This study was conducted to obtain basic data on the concentration and species distribution of fungi present in the indoor air of oak mushroom cultivation houses in Korea. In 2015 and 2016, we sampled and analyzed indoor airborne fungal spores 21 times from 13 oak mushroom cultivation farms located in six different provinces. The concentration of airborne fungi ranged from 1.30 × 102 to 1.59 × 104 cfu/m3. Surprisingly, in 20 sampling cases, the fungal concentration exceeded 500 cfu/m3, which is recommended as the indoor air quality standard by the Ministry of Environment, Korea. A total of 450 fungi were isolated and identified to belong to 33 genera and 46 species. Among the identified fungi, human pathogens (4 genera and 4 species) and plant pathogens (10 genera and 13 species) were present. In addition, Trichoderma harzianum, Trichoderma atroviride, and Trichoderma longibrachiatum, which are detrimental species that affect mushroom health, were found 17 out of 21 sampling times. Our results provide evidence that indoor air quality should be improved for better management of mushroom cultivation houses.