Antibiotic resistant bacteria survive treatment by doubling while shrinking

ABSTRACT Many antibiotics that are used in healthcare, farming, and aquaculture end up in environments with different spatial structures that might promote heterogeneity in the emergence of antibiotic resistance. However, the experimental evolution of microbes at sub-inhibitory concentrations of ant...

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Main Authors: Adrian Campey, Urszula Łapińska, Remy Chait, Krasimira Tsaneva-Atanasova, Stefano Pagliara
Format: Article
Language:English
Published: American Society for Microbiology 2024-12-01
Series:mBio
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Online Access:https://journals.asm.org/doi/10.1128/mbio.02375-24
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author Adrian Campey
Urszula Łapińska
Remy Chait
Krasimira Tsaneva-Atanasova
Stefano Pagliara
author_facet Adrian Campey
Urszula Łapińska
Remy Chait
Krasimira Tsaneva-Atanasova
Stefano Pagliara
author_sort Adrian Campey
collection DOAJ
description ABSTRACT Many antibiotics that are used in healthcare, farming, and aquaculture end up in environments with different spatial structures that might promote heterogeneity in the emergence of antibiotic resistance. However, the experimental evolution of microbes at sub-inhibitory concentrations of antibiotics has been mainly carried out at the population level which does not allow capturing single-cell responses to antibiotics. Here, we investigate and compare the emergence of resistance to ciprofloxacin in Escherichia coli in well-mixed and structured environments using experimental evolution, genomics, and microfluidics-based time-lapse microscopy. We discover that resistance to ciprofloxacin and cross-resistance to other antibiotics is stronger in the well-mixed environment due to the emergence of target mutations, whereas efflux regulator mutations emerge in the structured environment. The latter mutants also harbor sub-populations of persisters that survive high concentrations of ciprofloxacin that inhibit bacterial growth at the population level. In contrast, genetically resistant bacteria that display target mutations also survive high concentrations of ciprofloxacin that inhibit their growth via population-level antibiotic tolerance. These resistant and tolerant bacteria keep doubling while shrinking in size in the presence of ciprofloxacin and regain their original size after antibiotic removal, which constitutes a newly discovered phenotypic response. This new knowledge sheds light on the diversity of strategies employed by bacteria to survive antibiotics and poses a stepping stone for understanding the link between mutations at the population level and phenotypic single-cell responses.IMPORTANCEThe evolution of antimicrobial resistance poses a pressing challenge to global health with an estimated 5 million deaths associated with antimicrobial resistance every year globally. Here, we investigate the diversity of strategies employed by bacteria to survive antibiotics. We discovered that bacteria evolve genetic resistance to antibiotics while simultaneously displaying tolerance to very high doses of antibiotics by doubling while shrinking in size.
format Article
id doaj-art-bce3366e16e2425fa91cba48cf6a77a4
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issn 2150-7511
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spelling doaj-art-bce3366e16e2425fa91cba48cf6a77a42025-08-20T02:33:47ZengAmerican Society for MicrobiologymBio2150-75112024-12-01151210.1128/mbio.02375-24Antibiotic resistant bacteria survive treatment by doubling while shrinkingAdrian Campey0Urszula Łapińska1Remy Chait2Krasimira Tsaneva-Atanasova3Stefano Pagliara4Living Systems Institute and Biosciences, University of Exeter, Exeter, Devon, United KingdomLiving Systems Institute and Biosciences, University of Exeter, Exeter, Devon, United KingdomLiving Systems Institute and Biosciences, University of Exeter, Exeter, Devon, United KingdomLiving Systems Institute and Biosciences, University of Exeter, Exeter, Devon, United KingdomLiving Systems Institute and Biosciences, University of Exeter, Exeter, Devon, United KingdomABSTRACT Many antibiotics that are used in healthcare, farming, and aquaculture end up in environments with different spatial structures that might promote heterogeneity in the emergence of antibiotic resistance. However, the experimental evolution of microbes at sub-inhibitory concentrations of antibiotics has been mainly carried out at the population level which does not allow capturing single-cell responses to antibiotics. Here, we investigate and compare the emergence of resistance to ciprofloxacin in Escherichia coli in well-mixed and structured environments using experimental evolution, genomics, and microfluidics-based time-lapse microscopy. We discover that resistance to ciprofloxacin and cross-resistance to other antibiotics is stronger in the well-mixed environment due to the emergence of target mutations, whereas efflux regulator mutations emerge in the structured environment. The latter mutants also harbor sub-populations of persisters that survive high concentrations of ciprofloxacin that inhibit bacterial growth at the population level. In contrast, genetically resistant bacteria that display target mutations also survive high concentrations of ciprofloxacin that inhibit their growth via population-level antibiotic tolerance. These resistant and tolerant bacteria keep doubling while shrinking in size in the presence of ciprofloxacin and regain their original size after antibiotic removal, which constitutes a newly discovered phenotypic response. This new knowledge sheds light on the diversity of strategies employed by bacteria to survive antibiotics and poses a stepping stone for understanding the link between mutations at the population level and phenotypic single-cell responses.IMPORTANCEThe evolution of antimicrobial resistance poses a pressing challenge to global health with an estimated 5 million deaths associated with antimicrobial resistance every year globally. Here, we investigate the diversity of strategies employed by bacteria to survive antibiotics. We discovered that bacteria evolve genetic resistance to antibiotics while simultaneously displaying tolerance to very high doses of antibiotics by doubling while shrinking in size.https://journals.asm.org/doi/10.1128/mbio.02375-24environmental structureantimicrobial resistanceantibiotic toleranceevolutionmutationspersisters
spellingShingle Adrian Campey
Urszula Łapińska
Remy Chait
Krasimira Tsaneva-Atanasova
Stefano Pagliara
Antibiotic resistant bacteria survive treatment by doubling while shrinking
mBio
environmental structure
antimicrobial resistance
antibiotic tolerance
evolution
mutations
persisters
title Antibiotic resistant bacteria survive treatment by doubling while shrinking
title_full Antibiotic resistant bacteria survive treatment by doubling while shrinking
title_fullStr Antibiotic resistant bacteria survive treatment by doubling while shrinking
title_full_unstemmed Antibiotic resistant bacteria survive treatment by doubling while shrinking
title_short Antibiotic resistant bacteria survive treatment by doubling while shrinking
title_sort antibiotic resistant bacteria survive treatment by doubling while shrinking
topic environmental structure
antimicrobial resistance
antibiotic tolerance
evolution
mutations
persisters
url https://journals.asm.org/doi/10.1128/mbio.02375-24
work_keys_str_mv AT adriancampey antibioticresistantbacteriasurvivetreatmentbydoublingwhileshrinking
AT urszulałapinska antibioticresistantbacteriasurvivetreatmentbydoublingwhileshrinking
AT remychait antibioticresistantbacteriasurvivetreatmentbydoublingwhileshrinking
AT krasimiratsanevaatanasova antibioticresistantbacteriasurvivetreatmentbydoublingwhileshrinking
AT stefanopagliara antibioticresistantbacteriasurvivetreatmentbydoublingwhileshrinking