{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T06:39:36Z","timestamp":1792651176400,"version":"4.5.0"},"reference-count":44,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[2006,4,1]],"date-time":"2006-04-01T00:00:00Z","timestamp":1143849600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2006,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Legionella pneumophila<\/jats:italic>\n                    persists for a long time in aquatic habitats, where the bacteria associate with biofilms and replicate within protozoan predators. While\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    serves as a paradigm for intracellular growth within protozoa, it is less clear whether the bacteria form or replicate within biofilms in the absence of protozoa. In this study, we analyzed surface adherence of and biofilm formation by\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    in a rich medium that supported axenic replication. Biofilm formation by the virulent\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    strain JR32 and by clinical and environmental isolates was analyzed by confocal microscopy and crystal violet staining. Strain JR32 formed biofilms on glass surfaces and upright polystyrene wells, as well as on pins of \u201cinverse\u201d microtiter plates, indicating that biofilm formation was not simply due to sedimentation of the bacteria. Biofilm formation by an\n                    <jats:italic>L. pneumophila fliA<\/jats:italic>\n                    mutant lacking the alternative sigma factor \u03c3\n                    <jats:sup>28<\/jats:sup>\n                    was reduced, which demonstrated that bacterial factors are required. Accumulation of biomass coincided with an increase in the optical density at 600 nm and ceased when the bacteria reached the stationary growth phase.\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    neither grew nor formed biofilms in the inverse system if the medium was exchanged twice a day. However, after addition of\n                    <jats:italic>Acanthamoeba castellanii<\/jats:italic>\n                    , the bacteria proliferated and adhered to surfaces. Sessile (surface-attached) and planktonic (free-swimming)\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    expressed \u03b2-galactosidase activity to similar extents, and therefore, the observed lack of proliferation of surface-attached bacteria was not due to impaired protein synthesis or metabolic activity. Cocultivation of green fluorescent protein (GFP)- and DsRed-labeled\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    led to randomly interspersed cells on the substratum and in aggregates, and no sizeable patches of clonally growing bacteria were observed. Our findings indicate that biofilm formation by\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    in a rich medium is due to growth of planktonic bacteria rather than to growth of sessile bacteria. In agreement with this conclusion, GFP-labeled\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    initially adhered in a continuous-flow chamber system but detached over time; the detachment correlated with the flow rate, and there was no accumulation of biomass. Under these conditions,\n                    <jats:italic>L. pneumophila<\/jats:italic>\n                    persisted in biofilms formed by\n                    <jats:italic>Empedobacter breve<\/jats:italic>\n                    or\n                    <jats:italic>Microbacterium<\/jats:italic>\n                    sp. but not in biofilms formed by\n                    <jats:italic>Klebsiella pneumoniae<\/jats:italic>\n                    or other environmental bacteria, suggesting that specific interactions between the bacteria modulate adherence.\n                  <\/jats:p>","DOI":"10.1128\/aem.72.4.2885-2895.2006","type":"journal-article","created":{"date-parts":[[2006,4,6]],"date-time":"2006-04-06T09:43:49Z","timestamp":1144316629000},"page":"2885-2895","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":148,"title":["Planktonic Replication Is Essential for Biofilm Formation by\n                    <i>Legionella pneumophila<\/i>\n                    in a Complex Medium under Static and Dynamic Flow Conditions"],"prefix":"10.1128","volume":"72","author":[{"given":"Jo\u0308rg","family":"Mampel","sequence":"first","affiliation":[{"name":"Institute of Microbiology, Swiss Federal Institute of Technology (ETH), CH-8093 Zu\u0308rich, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Spirig","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Swiss Federal Institute of Technology (ETH), CH-8093 Zu\u0308rich, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan S.","family":"Weber","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Swiss Federal Institute of Technology (ETH), CH-8093 Zu\u0308rich, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Janus A. J.","family":"Haagensen","sequence":"additional","affiliation":[{"name":"Center for Biomedical Microbiology, Biocentrum-DTU, Technical University of Denmark, DK-2800 Lyngby, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S\u00f8ren","family":"Molin","sequence":"additional","affiliation":[{"name":"Center for Biomedical Microbiology, Biocentrum-DTU, Technical University of Denmark, DK-2800 Lyngby, Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hubert","family":"Hilbi","sequence":"additional","affiliation":[{"name":"Institute of Microbiology, Swiss Federal Institute of Technology (ETH), CH-8093 Zu\u0308rich, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1099\/mic.0.27563-0"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.37.6.1771-1776.1999"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1094226"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbrc.2005.04.060"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.69.4.2116-2122.2001"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.10.4.437-441.1979"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/0966-842X(96)10041-X"},{"key":"e_1_3_2_9_2","first-page":"135","year":"2002","unstructured":"Fields, B. 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