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. 2019 Aug 22;9:12252. doi: 10.1038/s41598-019-48697-x

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Comparison of biofilm formation by F. tularensis LVS, WbtI_G191V (LPS O-Ag mutant), LVSΔ1423-22 (CLC mutant), and O-Ag and CLC double mutant before and after passage in CDMB by confocal laser scanning microscopy. Panel (A) contains orthogonal sections showing horizontal (z) and side views (x and y) of reconstructed three-dimensional biofilm images at a magnification of 25x. Biofilms were stained with Syto9 showing both live and dead bacterial cells. Panel (B) shows topographical images of biofilm growth of the parent LVS and its surface structure mutant strains. Loss of surface carbohydrate was associated with denser biofilms, but subculture passage in CDMB resulted in thinner biofilms.