Figure 1. Biofilm susceptibility to antibiotic treatment

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Figure 1. Biofilm susceptibility to antibiotic treatment Figure 1. Biofilm susceptibility to antibiotic treatment. PAO1 biofilms grown in mucus or PBS for 24 h were treated ... Figure 1. Biofilm susceptibility to antibiotic treatment. PAO1 biofilms grown in mucus or PBS for 24 h were treated with tobramycin (a) or colistin (b). After 24 h of incubation, efficacy was assessed by determination of cfu. The cfu counts are depicted logarithmically as regression curves showing the mean ± SD for n = 3 experiments, each with technical duplicates. A double asterisk indicates statistical significance at P < 0.01, according to the Mann–Whitney U-test, for comparison of mucus versus PBS at the individual concentrations. The broken line indicates the detection limit. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) J Antimicrob Chemother, Volume 73, Issue 10, 05 July 2018, Pages 2762–2769, https://doi.org/10.1093/jac/dky241 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 2. Tobramycin activity in human mucus Figure 2. Tobramycin activity in human mucus. Mucus pre-incubated with tobramycin was inoculated with PAO1 and ... Figure 2. Tobramycin activity in human mucus. Mucus pre-incubated with tobramycin was inoculated with PAO1 and incubated for 24 h. Medium only and medium containing the same concentration of tobramycin were used as controls. Bar graphs show the mean + SD for n = 2 samples, each with technical duplicates. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) J Antimicrob Chemother, Volume 73, Issue 10, 05 July 2018, Pages 2762–2769, https://doi.org/10.1093/jac/dky241 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 3. Biofilm structure and bacterial count in mucus Figure 3. Biofilm structure and bacterial count in mucus. GFP-PAO1 biofilms were grown in mucus (a) or PBS (b) for ... Figure 3. Biofilm structure and bacterial count in mucus. GFP-PAO1 biofilms were grown in mucus (a) or PBS (b) for 2 days and imaged by confocal microscopy. Uninfected mucus was imaged as a control (c). Fluorescence signals were quantified using Imaris software (d) and cfu were determined (e). Bar graphs show the mean + SD for n = 3 samples, each with technical duplicates. A non-parametric t-test (Mann–Whitney U-test) was performed for statistical analyses. *P < 0.05. **P < 0.01. Scale bars represent 20 μm. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) J Antimicrob Chemother, Volume 73, Issue 10, 05 July 2018, Pages 2762–2769, https://doi.org/10.1093/jac/dky241 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 4. Tobramycin and colistin diffusion through biofilm formed in mucus. PAO1 biofilm cultured in mucus or PBS in ... Figure 4. Tobramycin and colistin diffusion through biofilm formed in mucus. PAO1 biofilm cultured in mucus or PBS in the apical compartment of a Transwell<sup>®</sup> was treated with tobramycin or colistin. PBS only or sham-infected mucus was used as a control. Diffusion of the antibiotic was assessed by HPLC and is depicted as cumulative mass. Scatter plots show the mean ± SEM for n = 3 experiments, each with triplicates. A single asterisk, a double asterisk and a triple asterisk indicate statistical significance at P < 0.05, P < 0.01 and P < 0.001, respectively, according to the Mann–Whitney U-test, for comparison of biofilm in mucus with PBS. A single hash, a double hash and a triple hash indicate statistical significance at P < 0.05, P < 0.01 and P < 0.001, respectively, according to the Mann–Whitney U-test, for comparison of biofilm in mucus with mucus only. A one-phase exponential association fit was performed for tobramycin diffusion using GraphPad Prism 7, described by the equation Y = Y<sub>max</sub> × (1 − e<sup>−</sup><sup>k</sup><sup>×</sup><sup>x</sup>). Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2018. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) J Antimicrob Chemother, Volume 73, Issue 10, 05 July 2018, Pages 2762–2769, https://doi.org/10.1093/jac/dky241 The content of this slide may be subject to copyright: please see the slide notes for details.