AbstractBacterial pathogenesis involves complex mechanisms contributing to virulence and persistence of infections. Understanding the multifactorial nature of bacterial infections is crucial for developing effective interventions. The present study investigated the efficacy of indole-3-acetic acid (IAA) against Pseudomonas aeruginosa with various end points including antibacterial activity, minimum inhibitory concentration (MIC), virulence factor production, biofilm inhibition, bacterial cell detachment, and viability assays. Results showed significant biofilm inhibition, bacterial cell detachment, and modest effects on bacterial viability. Microscopic analysis confirmed the disintegrated biofilm matrix, supporting the inhibitory effect of IAA. Additionally, molecular docking studies revealed potential mechanisms of action through active bond interactions between IAA and virulence proteins. These findings highlight IAA as an effective antibiofilm agent against P. aeruginosa.Keywords: Antibacterial compoundsantimicrobial resistancebacterial biofilmpathogenesisvirulence factors AcknowledgementsThe authors are thankful to UGC-SAP-DRS-II (F.3-9/2013[SAP-II], Department of Science and Technology Fund for Improvement of Science and Technology Infrastructure (DST-FIST) Level I (Stage II) (Ref. No. SR/FST/LSI-647/2015(C) Date.11.08.2016), and the Department of Science and Technology Promotion of University Research and Scientific Excellence (DST PURSE PhaseII) (Ref. No. SR/PURSE PHASE 2/16(G)/& 16(C) Date. 21.02.2017) of the Department of Animal Science, Bharathidasan University for the instrumentation facility: University Science Instrumentation Center (USIC), Bharathidasan University for providing instrumentation facilities (CLSM, SEM and Schrödinger). The authors also thank "RUSA, 2.0-Biological Sciences, Bharathidasan University".Data availability statementAll data analysed or generated throughout this study are included in this published article.Disclosure statementNo potential conflict of interest was reported by the author(s).Additional informationFundingThe author(s) reported there is no funding associated with the work featured in this article.