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They act by displacement and chelation, respectively, of divalent cations in bacterial membranes and may therefore act synergistically when applied in combination. If so, this combination of agents may be useful for the treatment of diseases like cystic fibrosis (CF), in which biofilms are present on the respiratory epithelium. We used checkerboard assays to investigate the synergy between these agents using reference strains\n            <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n            ATCC 27853 and\n            <jats:italic>Staphylococcus aureus<\/jats:italic>\n            ATCC 6538 in planktonic form. We then determined the efficacy of each agent against biofilms of both species grown on 96-pin lids and proceeded to combination testing against the\n            <jats:italic>P. aeruginosa<\/jats:italic>\n            reference strain and 10 clinical isolates from patients with CF. Synergism was observed for planktonic forms of both species and for biofilms of\n            <jats:italic>P. aeruginosa<\/jats:italic>\n            . The susceptibility of biofilms of\n            <jats:italic>P. aeruginosa<\/jats:italic>\n            clinical isolates to these agents was variable compared to the laboratory reference strain. This combination of agents may be useful in the management of biofilm-associated conditions, particularly those amenable to topical therapies. These results provide a basis upon which the antimicrobial and antibiofilm efficacy of preparations containing these agents may be enhanced.\n          <\/jats:p>\n          <jats:sec>\n            <jats:title>IMPORTANCE<\/jats:title>\n            <jats:p>\n              Bacteria living in biofilms produce a protective matrix which makes them difficult to kill. Patients with severe respiratory disease often have biofilms. Polymyxin B is an antibiotic commonly used in topical medications, such as eye drops and nasal sprays. Ethylenediaminetetraacetic acid (EDTA) is used widely as a preservative in medication but also has antimicrobial properties. It has been hypothesized that Polymyxin B and EDTA could have a synergistic relationship: when used in combination their antimicrobial effect is enhanced. Here, we evaluated the levels at which Polymyxin B and EDTA work together to kill common pathogens\n              <jats:italic>Pseudomonas aeruginosa<\/jats:italic>\n              and\n              <jats:italic>Staphylococcus aureus<\/jats:italic>\n              . We found that Polymyxin B and EDTA were synergistic. This synergy may be useful in the management of planktonic infection with\n              <jats:italic>P. aeruginosa<\/jats:italic>\n              and\n              <jats:italic>S. aureus<\/jats:italic>\n              , or biofilm infection with\n              <jats:italic>P. aeruginosa<\/jats:italic>\n              . This synergy may be beneficial in the treatment of respiratory biofilms, in which\n              <jats:italic>P. aeruginosa<\/jats:italic>\n              biofilms are common.\n            <\/jats:p>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.01709-23","type":"journal-article","created":{"date-parts":[[2024,1,3]],"date-time":"2024-01-03T14:01:06Z","timestamp":1704290466000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":14,"title":["Polymyxin B and ethylenediaminetetraacetic acid act synergistically against\n            <i>Pseudomonas aeruginosa<\/i>\n            and\n            <i>Staphylococcus aureus<\/i>"],"prefix":"10.1128","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5860-2448","authenticated-orcid":false,"given":"Samuel J. M.","family":"Hale","sequence":"first","affiliation":[{"name":"Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan J.","family":"Cameron","sequence":"additional","affiliation":[{"name":"School of Chemical Sciences and School of Biological Sciences, The University of Auckland, Auckland, New Zealand"},{"name":"Maurice Wilkins Centre for Molecular Biodiscovery, The University of Auckland, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian A.","family":"Lux","sequence":"additional","affiliation":[{"name":"Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristi","family":"Biswas","sequence":"additional","affiliation":[{"name":"Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raymond","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Surgery, Faculty of Medical and Health Sciences, The University of Auckland, Grafton, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"O'Carroll","sequence":"additional","affiliation":[{"name":"Respiratory Services, Auckland City Hospital, Te Toka Tumai, Te Whatu Ora, Auckland, New Zealand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul W. 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