Characterization and Treatment of Polymicrobial Biofilms in Prosthetic Joint Infections
Abstract
Abstract Prosthetic joint infections are complex and associated with high economic and social costs. Polymicrobial cases remain poorly understood, particularly regarding bacterial interactions and antibiotic response within mixed-species biofilms. To address these challenges, we introduce two approaches for characterizing polymicrobial biofilms: a modeling equation and fatty acid profiling. Predictive modeling estimates biofilm behavior. Fatty acid profiling infers species dominance and interspecies interactions. We integrate these results with fluorescence in situ hybridization and confocal microscopy. This provides a comprehensive view of biofilm structure and composition. Using this multi-modal approach, we characterize mono-, dual-, and triple-species biofilms formed by Klebsiella pneumoniae , Pseudomonas aeruginosa , and Staphylococcus aureus . In parallel, we systematically evaluate common anti-biofilm antibiotics, alone and in combination, across all biofilm types. The integrated analytical approaches yielded consistent findings. K. pneumoniae emerged as predominant in all biofilms where it was present. S. aureus and P. aeruginosa were in lower proportions and competed with each other. S. aureus dominated dual-species biofilms with P. aeruginosa . Antibiotic efficacy depended on biofilm composition and spatial organization. Among all pairs tested, colistin with levofloxacin showed the highest activity. Biofilm persistence was mainly driven by S. aureus. This species…
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