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This study was conducted to set up a proper replacement cycle of High Efficiency Particulate Air (HEPA) filters by observing the microbial populations in the air of the cultivation house of Pleurotus eryngii, before and after HEPA filter replacement at different periods . The density of bacteria and fungi in the air during each cultivation stage was measured using a sampler before the replacement of the HEPA filter. The results showed that airborne microorganisms had the highest density in the mushroom medium preparation room, with 169.7 CFU/m3 of bacteria and 570 CFU/m3 of fungi, and the removed old spaun had 126.3 CFU/m3 of bacteria and 560 CFU/m3 of fungi. The density of bacteria and fungi in the air at each cultivation stage before the replacement of the HEPA filter was 169.7 CFU/m3 and 570 CFU/m3, and 126.3 CFU/m3 and 560 CFU/m3, during the medium production and harvesting processes, respectively. After the replacement of the HEPA filter, the bacterial density was the lowest in the incubation room and the fungal density was the lowest in the cooling room. The microbial populations isolated at each period consisted of seven genera and seven species before the replacement, including Cladosporium sp., six genera and six species after 1 month of replacement, including Penicillium sp., 5 genera and 7 species after 3 months of replacement, including Mucor plumbeus, and 5 genera and 12 species, 5 genera and 10 species, and 5 genera and 10 species, 4, 5, and 6 months after the replacement, respectively, including Penicillium brevicompactum. During the period after replacement, the species were diversified and their number increased. The density of airborne microorganisms decreased drastically after the replacement of the HEPA filter. Its lowest value was recorded after 2 months of replacement, and it increased gradually afterwards, reaching a level similar to or higher than that of the pre-replacement period. Therefore, it was concluded that replacing the HEPA filter every 6 months is effective for reducing contamination.
