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Treatment of\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      infections is increasingly challenging due to multi-drug resistance, and biofilm formation during infection further increases antibiotic tolerance. Iron, which is sequestered by the host innate immune system, is also a key nutrient that is required for\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      biofilm formation. The iron-responsive PrrF small regulatory RNAs (sRNAs) are key to\n                      <jats:italic toggle=\"yes\">P. aeruginosa\u2019s<\/jats:italic>\n                      iron starvation response, promote the production of the\n                      <jats:italic toggle=\"yes\">Pseudomonas<\/jats:italic>\n                      quinolone signal (PQS) quorum sensing molecule, and are required for virulence in murine lung infection. Prior work showed that the PrrF sRNAs are dispensable for biofilm formation; however, these studies were performed using flow-cell biofilms grown at room temperature. Here, we demonstrate a temperature dependency for PrrF in\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      biofilm formation: the genes for these sRNAs are required for optimal biofilm formation at 37\u00b0C but not 25\u00b0C. We further show that a \u2206\n                      <jats:italic toggle=\"yes\">pqsA<\/jats:italic>\n                      mutant, which lacks production of PQS and related metabolites, phenocopies the \u2206\n                      <jats:italic toggle=\"yes\">prrF<\/jats:italic>\n                      mutant. These studies demonstrate the importance of the PrrF sRNAs in\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      biofilm formation at body temperature and reveal a previously underappreciated role of temperature in iron homeostasis and\n                      <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                      biofilm physiology.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>\n                        Biofilm formation is a critical virulence trait for many microbial pathogens that confers tolerance to the host immune system and antimicrobials.\n                        <jats:italic toggle=\"yes\">Pseudomonas aeruginosa<\/jats:italic>\n                        is an opportunistic pathogen that forms biofilms resulting in treatment failure. Iron is a known requirement for\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        biofilm formation, yet the precise role of iron in biofilm physiology remains unclear. Here, we show that temperature alters the requirement for the PrrF small regulatory RNAs, key components of\n                        <jats:italic toggle=\"yes\">P. aeruginosa\u2019s<\/jats:italic>\n                        iron starvation response, for biofilm formation. Specifically, PrrF is required for the optimal formation of flow-cell biofilms at 37\u00b0C but not at 25\u00b0C, yet most flow-cell biofilm studies are conducted at 25\u00b0C. These results demonstrate a previously underappreciated role of temperature in\n                        <jats:italic toggle=\"yes\">P. aeruginosa<\/jats:italic>\n                        biofilm physiology.\n                      <\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/jb.00507-25","type":"journal-article","created":{"date-parts":[[2026,1,30]],"date-time":"2026-01-30T21:05:58Z","timestamp":1769807158000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["The\n                    <i>Pseudomonas aeruginosa<\/i>\n                    PrrF sRNAs and PqsA promote biofilm formation at body temperature"],"prefix":"10.1128","volume":"208","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0941-9524","authenticated-orcid":true,"given":"Rhishita","family":"Chourashi","sequence":"first","affiliation":[{"name":"Department of Pharmaceutical Sciences, School of Pharmacy, 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