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Infections triggered by DTR\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      urgently require research and development of innovative antibiotics. \u03b2-Carotene is a carotenoid pigment with multiple activities, such as antibacterial, antifungal, antioxidant, and anticancer activities, therefore, the current study aimed to investigate the potential antipseudomonal activity of \u03b2-carotene against DTR\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      to find a new treatment option.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Antimicrobial susceptibility tests were conducted for 100\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      isolates. Well diffusion and broth microdilution techniques were utilized to determine the antibacterial activity of \u03b2-carotene. A total of 10 virulence factors (biofilm, pyocyanin, motility, protease, gelatinase, exopolysaccharide, siderophore, hemolysin, pyomelanin, and rhamnolipid) were screened among the tested isolates, and the effect of \u03b2-carotene on these virulence factors was assessed. Light microscopy and confocal laser scanning microscopy approaches were established to investigate the effect of \u03b2-carotene on biofilm formation by DTR isolates. Molecular analysis, docking study, and the wound infection model further evaluated the antipseudomonal action of \u03b2-carotene against DTR isolates.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      All tested multidrug-resistant (MDR) and DTR isolates (\n                      <jats:italic>n<\/jats:italic>\n                      \u2009=\u200940) were susceptible to \u03b2-carotene (200\u00a0\u00b5g\/ml), with inhibition zones ranging from 10 to 33\u00a0mm, recording minimum inhibitory concentration (MIC) values of 12.5 to 100\u00a0\u00b5g\/ml. It was reported that \u03b2-carotene reduced the production of all tested virulence factors by DTR isolates, and the percentages of inhibition ranged from 21.5 to 100%. Additionally, \u03b2-carotene-treated biofilms showed a significant reduction in biomass and thickness (up to 60%). Moreover, the production of virulence-associated genes, namely\n                      <jats:italic>lasR<\/jats:italic>\n                      ,\n                      <jats:italic>rhlR<\/jats:italic>\n                      , and\n                      <jats:italic>pqsR<\/jats:italic>\n                      genes, was downregulated by 0.5 MIC of \u03b2-carotene. The molecular docking displayed a resilient LasR-\u03b2-carotene complex with a binding free energy of -8.6\u00a0kcal\/mol. The \u03b2-carotene-treated rat group showed accelerated wound closure (up to 90% at day 6) and lower\n                      <jats:italic>Pseudomonas<\/jats:italic>\n                      burden (3.3 colony forming unit\/g). In addition, the histological examination showed re-epithelialization in the epidermis layer with a few capillaries and newly formed hair follicles in the treated group.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>\n                      This is the first study to report the antibacterial, antivirulence, and antiquorum sensing activities of \u03b2-carotene against DTR\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      , signifying the promising role of \u03b2-carotene in mitigating DTR\n                      <jats:italic>P. aeruginosa<\/jats:italic>\n                      -caused infections.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12866-025-04234-7","type":"journal-article","created":{"date-parts":[[2025,8,12]],"date-time":"2025-08-12T02:47:22Z","timestamp":1754966842000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["In vitro and in vivo investigation of the antibacterial, antivirulence, and antiquorum sensing activities of \u03b2-carotene against difficult-to-treat resistant Pseudomonas aeruginosa"],"prefix":"10.1186","volume":"25","author":[{"given":"Khaled B.","family":"Al-Monofy","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ahmed A.","family":"Abdelaziz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amal M.","family":"Abo-Kamar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lamiaa A.","family":"Al-Madboly","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mahmoud H.","family":"Farghali","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,8,12]]},"reference":[{"key":"4234_CR1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2024.1413434","volume":"15","author":"L Cao","year":"2024","unstructured":"Cao L, Yang H, Huang Z, Lu C, Chen F, Zhang J, Zhang H. 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