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Background: Fungi are common in indoor environments and can cause various diseases. This study aimed to evaluate the extent of fungal contamination in hospital rooms and to assess the susceptibility of mold species isolated from indoor air to cycloheximide. Methods: A prospective cross-sectional study was conducted from November 2023 to February 2024 in five departments of the Sourô Sanou Teaching Hospital, Burkina Faso. Fifty-seven indoor air samples were collected using the passive sedimentation method. The samples were incubated at 30 °C for 24 h, and the resulting colonies were purified. Fungal identification was performed based on macroscopic and microscopic characteristics, and the isolated molds were tested for susceptibility to cycloheximide (0.4 mg/mL). Results: Overall, fungi were recovered from 70.2% (40/57) of air samples, with molds representing the majority of the fungal isolates (99.1%, 224/226). Among the 224 mold isolates identified, A. niger was the most prevalent species (48.7%, 109/224), followed by A. flavus (14.7%, 33/224), Rhizopus spp. (12.5%, 28/224), and A. fumigatus (6.7%, 15/224). The overall cycloheximide inhibition rate was 71.4% (160/224). Lower inhibition rates were observed for A. fumigatus (46.7%), A. versicolor (33.3%), A. flavus (27.3%), Penicillium spp. (27.3%), whereas A. terreus showed complete resistance. Conclusions: These findings underscore the importance of environmental monitoring in preventing and controlling hospital-acquired fungal infections. They also indicate that molds are not uniformly inhibited by cycloheximide, which may influence the accuracy of culture-based diagnostics.

Keywords: ; hospital environment; mold species; cycloheximide susceptibility; Burkina Faso