Attenuation of Staphylococcus aureus Biofilms and Virulence by 3-Fluorocatechol

<b>Background/Objectives:</b><i>Staphylococcus aureus</i> is a well-known opportunistic pathogen that causes a wide range of infections, from cutaneous blemishes to potentially fatal systemic diseases. The increasing prevalence of antibiotic-resistant bacteria highlights the critical need for alternative therapeutic methods that target virulence factors rather than growth. <b>Methods:</b> The antibacterial activity of 3-fluorocatechol (3-FC) against bacterial and fungal pathogens (e.g., <i>Candida albicans</i>) was determined by broth microdilution to establish the lowest inhibitory concentration. The antibiofilm impact of 3-FC against <i>S. aureus</i> was evaluated using crystal violet staining and viable colony counts, followed by scanning electron microscopy to visualize the biofilm architecture. The methanol extraction method was used to quantify staphyloxanthin synthesis in <i>S. aureus</i> cells. Furthermore, in silico molecular docking was used to evaluate 3-FC binding interactions and provide mechanistic insight into its impacts on <i>S. aureus</i> biofilms and virulence-associated factors. <b>Results:</b> Although the study showed that 3-FC exhibits weak antibacterial activity against <i>S. aureus</i> (MIC > 2048 µg/mL), it shows effective inhibition of up to 86.5% at sub-inhibitory doses during the initial stage of biofilm formation. The CFU enumeration also confirms the significant reduction of viable cell count of <i>S. aureus</i> in the presence of sub-MIC of 3-FC. The SEM analysis confirms disruption of the <i>S. aureus</i> biofilm architecture in the presence of a sub-MIC of 3-FC. Furthermore, the eradication of mature <i>S. aureus</i> biofilm at a sub-MIC dose of 3-FC was 60.6%. 3-FC significantly reduced staphyloxanthin formation, a vital antioxidant pigment that contributes to bacterial pathogenicity, with a maximal suppression of 66.3% at 2048 µg/mL. Molecular docking analyses provide further insight into the molecular basis of 3-FC activity, revealing strong binding affinities with numerous <i>S. aureus</i> virulence regulators and enzymes, suggesting interference with quorum-sensing, adhesion, and oxidative-stress response pathways. <b>Conclusions:</b> Collectively, our findings indicate that 3-FC has antibiofilm and antivirulence properties against <i>S. aureus.</i> Furthermore, this study suggests 3-FC as a viable structural scaffold for the development of a novel anti-infective agent to treat chronic staphylococcal infections.