Investigating the effect of several factors on concentrations of bioaerosols in a well-ventilated hospital environment Abstract This study characterized and quantified the bacterial and fungal bioaerosols in nine wards of the Razavi Hospital (Mashhad, Iran) that is equipped with an advanced heating, ventilating, and air conditioning (HVAC) system including HEPA filters for air cleaning. In this study, 432 samples were taken from the indoor air of multiple hospital wards during the morning and afternoon shifts during summer and autumn. The particle number concentrations with sizes of > 0.3, > 0.5, > 1, > 2, > 5, and > 10 μm were measured using a 6-channel handheld particle counter. A greater diversity of bioaerosol types were observed during the morning shifts and during summer. The microbial load was not affected significantly by the temperature, relative humidity, working shift, season, and number of visitors, indicating the effectiveness of a well-designed ventilation system to eliminate site-specific variations. For microbial number concentrations, a significant correlation was only observed between the number of particles with a diameter of > 10 μm and the airborne microbial loading. Thus, passive sampling may not properly reflect the actual concentrations of smaller bioaerosols. In conclusion, HEPA filters in the HVAC system successfully decreased the bioaerosol concentrations in the hospital environment. Additionally, we recommend that active sampling be used in cases where a well-functioning HVAC system exists.