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. 2021 Jun 24;11:13230. doi: 10.1038/s41598-021-92718-7

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Effect of bicarbonate on colistin resistance. (a) Bicarbonate sensitivity of B. thailandensis ∆dbcA. Spot assay of B. thailandensis strains (E264; vec, E264; DbcA, ∆dbcA; vec, and ∆dbcA; DbcA) on MH2 agar media with 100 µg/ml Tmp, 0.002% rhamnose, and different concentrations of sodium bicarbonate as indicated. (b) Reduction in colistin MIC of B. thailandensis E264 by bicarbonate. The MIC was determined on MH2 agar plates with different concentrations of sodium bicarbonate as indicated. Approximate MICs are denoted by white arrows. Plates were analyzed after 48 h of growth at 37 °C. (c) Bicarbonate sensitivity of E. coli ∆yqjA and complementation of bicarbonate sensitivity of E. coli ∆yqjA by B. thailandensis dbcA. Spot assay of 1:10 dilutions of E. coli strains (W3110; vec, ∆yqjA; vec, ∆yqjA; YqjA, and ∆yqjA; DbcA) on LB agar media with 50 µg/ml Kan and 20 mM sodium bicarbonate as indicated. (d) Bicarbonate sensitivity of E. coli ∆nhaA. Spot assay of 1:10 dilutions of indicated strains on LB agar plate with and without 25 mM sodium bicarbonate. (e) Effect of bicarbonate on colistin MIC of different bacterial species. MIC was determined for indicated strains on LB agar plates with and without physiological concentration of bicarbonate (~ 25 mM). The pH of growth media was adjusted to pH 7.0 to avoid external pH fluctuations by sodium bicarbonate. Approximate MICs are denoted by white arrows.