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Using a nominally macrolide-resistant model <jats:italic>Klebsiella pneumoniae<\/jats:italic> strain (ATCC 10031) we demonstrate the effectiveness of several macrolides in inhibiting biofilm growth in multi-well plates, and the ability of azithromycin (AZM) to improve the effectiveness of the antibacterial last-agent-of-choice for <jats:italic>K<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic> infections, colistin methanesulfonate (CMS), against biofilms. This synergistic action was also seen in biofilm tests of several <jats:italic>K<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic> hospital isolates and could also be identified in polymyxin B disc-diffusion assays on azithromycin plates. Our work highlights the complexity of antimicrobial-resistance in bacterial pathogens and the need to test antibiotics with biofilm models where potential synergies might provide new therapeutic opportunities not seen in liquid culture or colony-based assays.<\/jats:p>","DOI":"10.1371\/journal.pone.0270983","type":"journal-article","created":{"date-parts":[[2022,7,1]],"date-time":"2022-07-01T17:44:38Z","timestamp":1656697478000},"page":"e0270983","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":14,"title":["Azithromycin possesses biofilm\u2013inhibitory activity and potentiates non-bactericidal colistin methanesulfonate (CMS) and polymyxin B against Klebsiella pneumonia"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2209-8681","authenticated-orcid":true,"given":"Olena V.","family":"Moshynets","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9248-4097","authenticated-orcid":true,"given":"Taras P.","family":"Baranovskyi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Scott","family":"Cameron","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8876-6075","authenticated-orcid":true,"given":"Olga S.","family":"Iungin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ianina","family":"Pokholenko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robyn","family":"Jerdan","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aleksandr","family":"Kamyshnyi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexey A.","family":"Krikunov","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4927-7946","authenticated-orcid":true,"given":"Viktoria V.","family":"Potochilova","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7834-233X","authenticated-orcid":true,"given":"Kateryna L.","family":"Rudnieva","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0463-8629","authenticated-orcid":true,"given":"Andrew J.","family":"Spiers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2022,7,1]]},"reference":[{"key":"pone.0270983.ref001","first-page":"110","article-title":"Antibiotic resistance in the current realities of cardiac surgery","volume":"1","author":"VV Lazoryshynetz","year":"2017","journal-title":"Ukrainian J Cardiovascular Surgery"},{"key":"pone.0270983.ref002","first-page":"97","article-title":"Spectrum of pathogens causing purulent inflammatory diseases in surgical patients","volume":"1","author":"GA Loban","year":"2021","journal-title":"Actualni problemi suchanoi medetsini: Vistnyk ukrainskoi medichnoi akademii"},{"key":"pone.0270983.ref003","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1093\/cid\/cis588","article-title":"Predictors of mortality in bloodstream infections caused by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae: importance of combination therapy","volume":"55","author":"M Tumbarello","year":"2012","journal-title":"Clin Infect Dis"},{"key":"pone.0270983.ref004","doi-asserted-by":"crossref","first-page":"30062","DOI":"10.2807\/1560-7917.ES.2015.20.45.30062","article-title":"the European Survey of Carbapenemase-Producing Enterobacteriaceae (EuSCAPE) working group. 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