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. 2018 Jun 5;8:8617. doi: 10.1038/s41598-018-26974-5

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© The Author(s) 2018

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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Effects of terrein on virulence factor production and biofilm formation. (a) Chemical structures of terrein and furanone C-30. (b) Effects of terrein on virulence factor production and cell viability in P. aeruginosa PAO1. PAO1 cells were grown in LB medium containing various concentrations of terrein for 24 h, followed by the measurement of cell density at 600 nm, elastase activity, pyocyanin, and rhamnolipid in the culture supernatants. (c) Effects of terrein on biofilm formation and cell viability in P. aeruginosa. P. aeruginosa biofilms formed in the presence of terrein for 9 h. The biofilm cells attached to the well surface were assayed using crystal violet staining. (d) Effects of terrein on the antibiotic tolerance of biofilms. Preformed biofilms (6-h) of PAO1 cells were treated with ciprofloxacin (Cip) alone or together with DMSO or 300 μM terrein for 3 h. The biofilms were dissociated from the wells by gentle sonication, and the bacteria were enumerated by plating. Three independent experiments were performed in triplicate, and the mean ± standard deviation (SD) values are displayed in each bar. *P < 0.01; **P < 0.001; ***P < 0.0001 versus untreated cells.