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. 2020 Jun 2;10:9021. doi: 10.1038/s41598-020-66082-x

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© The Author(s) 2020

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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Confocal images of inhibited S. aureus biofilms. (a) combined channel; (b) SYTO 9 channel; (c) Promidium Iodide channel; (1) control biofilm; (2) inhibited with 40 mM D-AA (40 mM D-Asp and 40 mM D-Glu); (3) treated with Cip; (4) treated with 40 mM D-AA and 8 MLC Cip; blue arrow) presence of intercellular eDNA; yellow arrow) lack of intercellular eDNA. (1) Untreated S. aureus biofilms are reinforced by an organised honeycomb like meshwork made of interconnected intercellular eDNA. eDNA forms a filamentous mesh like structure within the biofilm and surrounds all cells. (2) Biofilms inhibited with D-AA show a complete lack of eDNA. (3) Biofilms inhibited by Cip express a lower population of S. aureus cells. A dense eDNA meshwork provides structure to these persisting cells. (4) Biofilm treated with a combination of D-AA and 8 MLC Cip lack in eDNA meshwork structure; D-AA = 40 mM D-Aspartic acid and 40 mM D-Glutamic acid; x MLC = x times minimum lethal concentration; Cip = ciprofloxacin.