Biofilm Formation in Clinical Isolates of .

Many microbial pathogens form biofilms, assemblages of polymeric compounds that play a crucial role in establishing infections. The biofilms of species also contribute to high antifungal resistance. Using our collection of 29 clinical isolates, we focused on characterizing differences in thermotolerance, anaerobic growth, and biofilm formation across four species complexes commonly found in clinical settings. We investigated the role of carbon sources, temperature, and fungal morphology on biofilm development. Using fluorescence microscopy, we followed the stages of biofilm formation. Biofilms were screened for sensitivity/resistance to the antifungals voriconazole (VOR), amphotericin B (AmB), and 5-fluorocytosine (5-FC). Our findings revealed generally poor thermotolerance and growth under anaerobic conditions across all species. VOR was more effective than AmB in controlling biofilm formation, but the combination of VOR, AmB, and 5-FC significantly reduced biofilm formation across all species. Additionally, biofilm formation varied under non-glucose carbon sources, highlighting the species' adaptability to different nutrient environments. Notably, early stage biofilms were primarily composed of lipids, while polysaccharides became dominant in late-stage biofilms, suggesting a dynamic shift in biofilm composition over time.