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It is an emergent need to find out new molecules to combat such infections. In the present study, we analyzed the antibacterial effect of pimozide (PMZ) against gram\u2010positive and gram\u2010negative bacterial strains, including methicillin\u2010sensitive (MSSA) and methicillin\u2010resistant (MRSA) <jats:italic>S. aureus<\/jats:italic>. The growth of MSSA and MRSA was completely inhibited at concentrations of 12.5 and 100\u00a0\u03bcg\/mL, respectively, which is referred to as 1\u00d7 minimum inhibitory concentration (MIC). The cell viability was completely eliminated within 90\u00a0min of PMZ treatment (2\u00d7 MIC) through reactive oxygen species (ROS)\u2010mediated killing without affecting cell membrane permeability. It suppressed \u03b1\u2010hemolysin production and biofilm formation of different <jats:italic>S. aureus<\/jats:italic> strains by almost 50% at 1\u00d7 MIC concentration, and was found to detach matured biofilm. PMZ treatment effectively eliminates <jats:italic>S. aureus<\/jats:italic> infection in <jats:italic>Caenorhabditis elegans<\/jats:italic> and improves its survival by 90% and is found safe to use with no hemolytic effect on human and chicken blood tissues. Taken together, it is concluded that PMZ may turn out to be an effective antibacterial for treating bacterial infections including MSSA and MRSA.<\/jats:p>","DOI":"10.1002\/bab.2465","type":"journal-article","created":{"date-parts":[[2023,3,31]],"date-time":"2023-03-31T16:29:31Z","timestamp":1680280171000},"page":"1679-1689","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Antimicrobial effect of pimozide by targeting ROS\u2010mediated killing in <i>Staphylococcus aureus<\/i>"],"prefix":"10.1002","volume":"70","author":[{"given":"Siddhartha","family":"Kumar","sequence":"first","affiliation":[{"name":"Department of Biotechnology Central University of South Bihar  Gaya Bihar India"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kumar","family":"Sandeep","sequence":"additional","affiliation":[{"name":"Dr. B.R. 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