Airborne bacteria are expected to float in the mushroom cultivation house, as it is a special environment with high humidity and high temperatures. Their concentration and diversity in the indoor air of the cultivation house could effect the health of farmers and the quality of mushrooms. To examine whether microbiota of airborne bacteria change from year to year, we measured the indoor temperature, humidity, and airborne bacterial concentration from mushroom cultivation houses located in six regions in Korea from 2020 to 2021, and isolated and identified airborne bacteria. The surveyed data were compared and the bacterial diversity of the 1st year and the 2nd year were determined. Based on the average temperature and humidity data surveyed, it can be seen that the temperature and humidity environment in the cultivation houses is such that bacteria can easily reproduce. It was observed that the temperature inside the cultivation houses tends to be higher or lower depending on the season and correlates with the temperature outside the cultivation houses. In the first year survey, 32 species of 20 genera were identified, and in the second year survey, bacteria belonging to 29 species of 22 genera were identified. Among them, the most detected species were all species belonging to the genus Bacillus. There were only three species (Bacillus altitudinis, Brevibacterium frigoritolerans, and Staphylococcus epidermidis) that were continuously isolated in common. Our results showed that the species of floating bacteria greatly vary from year to year even for the same cultivation houses.