Aspergillus spp. in Woodworking Settings: Implications for Occupational Health and Safety

• Aspergillus section Fumigati was consistently present in woodworking environments. • Azole‑resistant Aspergillus fumigatus were found in DIY store. • Several isolates produced gliotoxin and showed cytotoxicity. • Warm seasons favoured Aspergillus section Fumigati growth in carpentries. • Findings support targeted measures to reduce fungal exposure. Woodworkers are exposed to several potentially harmful agents, including microorganisms that grow in the wood. The most common fungal species in woodworking environments are Aspergillus, Penicillium , and Cladosporium spp. with occupational exposure to Aspergillus spp. posing a significant respiratory risk. This study aimed to assess exposure to Aspergillus spp. in two Portuguese woodworking environments and to perform a thorough analysis of Aspergillus fumigatus complex isolates collected from 13 DIY stores and 6 Carpentries in Lisbon Metropolitan Area. Sampling combined active and passive methods to assess microbial contamination. Aspergillus fumigatus isolates were analysed for their antifungal susceptibility, resistant mechanisms, mycotoxins production and cytotoxic potential in lung (A459) and liver (HepG2) cell lines. The MAS-100 presented Aspergillus sections Aspergilli and Flavi with the highest prevalence in DIY stores and Carpentries, respectively. A total of 1185 Aspergillus spp. were recovered, 270 identified as Aspergillus fumigatus sensu stricto growing at 37°C. None of those isolates was resistant to azoles, 99.07% of them produced gliotoxin and 39.9% of them produced cytotoxic effects in at least one cell line. This study comprehended a multi-approach that considered not only sampling methods but also the laboratory assays to be applied in the Aspergillus section Fumigati isolates recovered from two different woodworking environments, allowing a complete and robust analysis of this specific environment and species. Overall, the findings indicate that woodworkers are exposed to A. fumigatus isolates with relevant pathogenic traits, despite the absence of azole resistance, underscoring the need for continued environmental and occupational monitoring.