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. 2020 Jun 10;10:9390. doi: 10.1038/s41598-020-66293-2

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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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Biofilm degradation activity. (A) The histogram shows the ability of different enzymes to degrade biofilms formed by P. aeruginosa PAO1 wt (solid bars) and PAO1ΔmucA (striped bars). The concentration of enzyme was the same in each sample (7.09 pmol/µl), that corresponds to 0.32 mg/ml for Alg2A; 0.14 mg/ml for A1-II; 0.22 mg/ml for A1-II’; and 0.23 mg/ml for A1-III. The values are normalized against the control sample (non-treated biofilm) that has 100% biomass. The decrease in the percentage of biomass reflects the effect of the treatment. The asterisk (*) indicates a significant difference (p < 0.05) when the sample is compared with the control. The (&) indicates a significant difference (p < 0.05) when the PAO1 wt strain is compared with PAO1ΔmucA. (B) Synergism between antibiotics and the AlyA1, A1-III and Alg2A alginate lyase proteins. The asterisk (*) indicates a significant difference (p < 0.05) when the sample is compared with the control. The (&) indicates a significant difference (p < 0.05) when alginase is compared with alginase + ciprofloxacin. The (∞) indicates a significant difference (p < 0.05) when the sample is compared with ciprofloxacin. (C) Sum of stack images and the corresponding orthogonal views of the confocal microscopy images of P. aeruginosa PAO1 wt and PAO1 ΔmucA biofilms treated with 7.09 pmol/µl of the different alginate lyases and 1 µg/ml of ciprofloxacin (CPX). Red and green colors show the P. aeruginosa dead and alive cells detected by the LIVE/DEAD staining kit, respectively. Scale bar corresponds to 50 µm.