Survival of Escherichia coli after high-antibiotic stress is dependent on both the pregrown physiological state and incubation conditions

Frontiers in Microbiology

Microbiology

Gaute Hovde Bø¹  Per Bruheim¹  Lilja Brekke Thorfinnsdottir¹  James Alexander Booth² 

[1] Department of Biotechnology and Food Science, Norwegian University of Science and Technology, Trondheim, Norway;Department of Microbiology, Oslo University Hospital, University of Oslo, Oslo, Norway;

关键词: persisters;    antibiotics;    metabolomics;    metabolite profiling;    Escherichia coli;    relA;    bioreactors;   

DOI  :  10.3389/fmicb.2023.1149978

 received in 2023-01-23, accepted in 2023-02-17,  发布年份 2023

来源: Frontiers

【 摘 要 】

IntroductionThe survival of bacterial cells exposed to antibiotics depends on the mode of action, the antibiotics concentration, and the duration of treatment. However, it also depends on the physiological state of the cells and the environmental conditions. In addition, bacterial cultures contain sub-populations that can survive high antibiotic concentrations, so-called persisters. Research on persisters is challenging due to multiple mechanisms for their formation and low fractions, down to and below one millionth of the total cell population. Here, we present an improved version of the persister assay used to enumerate the amount of persisters in a cell population.MethodsThe persister assay with high antibiotic stress exposure was performed at both growth supporting and non-supporting conditions. Escherichia coli cells were pregrown to various growth stages in shake flasks and bench-top bioreactors. In addition, the physiological state of E. coli before antibiotic treatment was determined by quantitative mass spectrometry-based metabolite profiling.ResultsSurvival of E. coli strongly depended on whether the persister assay medium supported growth or not. The results were also highly dependent on the type of antibiotic and pregrown physiological state of the cells. Therefore, applying the same conditions is critical for consistent and comparable results. No direct connection was observed between antibiotic efficacy to the metabolic state. This also includes the energetic state (i.e., the intracellular concentration of ATP and the adenylate energy charge), which has earlier been hypothesized to be decisive for persister formation.DiscussionThe study provides guides and suggestions for the design of future experimentation in the research fields of persisters and antibiotic tolerance.

【 授权许可】

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Copyright © 2023 Thorfinnsdottir, Bø, Booth and Bruheim.