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This ongoing problem has highlighted the question of whether disinfection recommendations for medical devices containing water circuits are adequate for preventing contamination and possible recontamination by nontuberculous mycobacteria. The formation of biofilms in such devices exacerbates the problem. This study aimed to assess the efficacy of disinfectants on biofilms and suspensions of the\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      strain ZUERICH-1, and to compare it with two unrelated\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      strains obtained from different sources.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      Disinfection efficacy testing for biofilm was performed using a Bead Assay for Biofilms and for bacteria in suspension according to the European Standard EN 14348. Three different disinfectants, glutaraldehyde, sodium hypochlorite and peracetic acid, were assessed.\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      ZUERICH-1, two genetically unrelated\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      isolates and\n                      <jats:italic>M. avium<\/jats:italic>\n                      subsp.\n                      <jats:italic>avium<\/jats:italic>\n                      ATCC 15769, which is included in European standards for disinfectant testing on mycobacteria, were analyzed. The biofilms\u2019 structure and composition were analyzed by chemical and molecular techniques and advanced imaging methods.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      We found that peracetic acid and glutaraldehyde in standard concentrations were able to effectively inactivate (\u2265\u20094 log\n                      <jats:sub>10<\/jats:sub>\n                      reduction) suspended bacteria of all three strains, but chlorine failed in all cases. Formation of biofilm generally enhanced the tolerance of\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      to disinfectants. Peracetic acid in standard concentration could not effectively inactivate biofilms of\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      ZUERICH-1, but was effective against biofilms of the other\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      strains tested. Similarly, glutaraldehyde in standard concentration could not inactivate biofilm of ZUERICH-1. Biomass analysis showed higher amounts of extracellular matrix of ZUERICH-1 when compared to the other two strains.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      The data suggest that current standard disinfection recommendations do not ensure sustained inhibition of\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      when embedded in biofilm. Additional measures are needed to prevent nosocomial transmission of\n                      <jats:italic>M. chimaera<\/jats:italic>\n                      through contaminated heater-cooler units.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12866-025-04439-w","type":"journal-article","created":{"date-parts":[[2025,11,13]],"date-time":"2025-11-13T06:54:14Z","timestamp":1763016854000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Biofilm formation by the global outbreak strain of Mycobacterium chimaera results in significantly reduced efficacy of standard disinfectants"],"prefix":"10.1186","volume":"25","author":[{"given":"Anna Maria","family":"Oschmann","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharina","family":"Konrat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christoph","family":"Schaudinn","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Genevieve","family":"Sohl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dirk","family":"Wagner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Astrid","family":"Lewin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7664-5150","authenticated-orcid":false,"given":"Mardjan","family":"Arvand","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,11,13]]},"reference":[{"key":"4439_CR1","doi-asserted-by":"crossref","unstructured":"Holton J. 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