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. 2020 Jan 15;10:3006. doi: 10.3389/fmicb.2019.03006

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Copyright © 2020 Bair and Campagnari.

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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Time course assay to assess the effects of lower seeding concentrations and address strain specificity by analyzing trends of co-infection isolates. Bacteria seeded simultaneously at a lower concentration (∼10⁵ CFU per mL) and maintained with media changes (black arrows) for 48 h in monomicrobial (dashed) or triple species (solid) biofilms. Each symbol and error bar represent the mean CFU per mL and standard deviation (SD) of biofilms at the respective time point. Data presented are from three independent assays with a minimum of two biologic replicates per assay. Black dashed line indicates the limit of CFU detection. (A) NTHi86-028 NP (blue), M. catarrhalis 7169 (green), and S. pneumoniae EF3030 (red) monomicrobial and triple species biofilms. (B) NTHi 11-01-125 (blue), M. catarrhalis 11-01-125 (green) and S. pneumoniae 11-01-125 (red) monomicrobial and triple species biofilms.