Targeting Uropathogenic Virulence and Biofilms Using Plant-Derived Extracts: A Comprehensive Analysis of Origanum vulgare Bioactivities.

Urinary tract infections (UTIs) pose a significant clinical challenge, primarily driven by uropathogenic bacteria such as Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa, whose persistence is facilitated by virulence factor expression, biofilm formation, and escalating antimicrobial resistance. In this study, methanolic, ethanolic, and aqueous extracts of Urtica dioica, Azadirachta indica, and Origanum vulgare were evaluated against sixteen clinically isolated uropathogens to assess their antibacterial and virulence-attenuating potential. Among the tested preparations, ethanolic extracts—particularly O. vulgare—exhibited the strongest activity, with minimum inhibitory concentrations ranging from 0.781 to 3.12 mg/mL. In addition to inhibiting growth, the extracts significantly suppressed key uropathogenic traits, including protease and siderophore production, bacterial motility, and the development of both early and mature biofilms. Phytochemical profiling of the most active extract by GC–MS and FTIR revealed diverse bioactive constituents consistent with its activity. Importantly, hemolytic evaluation using human erythrocytes demonstrated minimal cytotoxicity, supporting host compatibility. Collectively, these findings highlight virulence and biofilm attenuation as viable antimicrobial-sparing strategies and identify O. vulgare extract as a promising adjunctive candidate for mitigating biofilm-associated UTIs.