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. 2019 Aug 23;10:1949. doi: 10.3389/fmicb.2019.01949

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Copyright © 2019 Olsen, Røder, Russel, Madsen, Sørensen and Burmølle.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Effect of individual-species pre-colonization on biofilm formation and species composition. Biofilm formation in the Calgary assay was assessed by crystal violet staining after 24 h of growth in 1/4 TSB (A) and full strength TSB (B). Mono-species biofilms and biofilms pre-inoculated with TSB or all strains were included as controls. Dunnett’s tests were conducted with PI-TSB as control (single strains were not included), which only showed significant differences between the two controls (p < 0.05^∗) in biofilms grown for 24 h (A). No significant differences were observed in (B). Bars represent means of three biological replicates and dots represent means of four technical replicates (n = 3). (C) Species enumeration for each pre-inoculation experiment from the DFR. All CFU counts were grouped according to species. Dunnett’s tests were performed within each group with PI-TSB as control. Significant differences between the PI-TSB control and the PI experiments are shown. Bars represent means of three biological replicates and dots represent means of two technical replicates (n = 3) (p < 0.05^∗, p < 0.001^∗∗∗).