Rapid decline of bacterial drug-resistance in an antibiotic-free environment through phenotypic reversion

  1. Anett Dunai
  2. Reka Spohn
  3. Zoltán Farkas
  4. Viktoria Lazar
  5. Adam Gyorkei
  6. Gabor Apjok
  7. Gabor Boross
  8. Balazs Szappanos
  9. Gabor Grezal
  10. Aniko Farago
  11. Laszlo Bodai
  12. Balazs Papp
  13. Csaba Pal  Is a corresponding author
  1. Biological Research Centre of the Hungarian Academy of Sciences, Hungary
  2. University of Szeged, Hungary

Abstract

Antibiotic resistance typically induces a fitness cost that shapes the fate of antibiotic-resistant bacterial populations. However, the cost of resistance can be mitigated by compensatory mutations elsewhere in the genome, and therefore the loss of resistance may proceed too slowly to be of practical importance. We present our study on the efficacy and phenotypic impact of compensatory evolution in Escherichia coli strains carrying multiple resistance mutations. We have demonstrated that drug-resistance frequently declines within 480 generations during exposure to an antibiotic-free environment. The extent of resistance loss was found to be generally antibiotic-specific, driven by mutations that reduce both resistance level and fitness costs of antibiotic-resistance mutations. We conclude that phenotypic reversion to the antibiotic-sensitive state can be mediated by the acquisition of additional mutations, while maintaining the original resistance mutations. Our study indicates that restricting antimicrobial usage could be a useful policy, but for certain antibiotics only.

Data availability

Sequencing data have been deposited in the NCBI Sequence Read Archive (SRA) under the accession number of PRJNA529335.

The following data sets were generated

Article and author information

Author details

  1. Anett Dunai

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  2. Reka Spohn

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  3. Zoltán Farkas

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5085-3306
  4. Viktoria Lazar

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  5. Adam Gyorkei

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  6. Gabor Apjok

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  7. Gabor Boross

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7208-5678
  8. Balazs Szappanos

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  9. Gabor Grezal

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  10. Aniko Farago

    Faculty of Science and Informatics, University of Szeged, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  11. Laszlo Bodai

    Department of Biochemistry and Molecular Biology, University of Szeged, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8411-626X
  12. Balazs Papp

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    Competing interests
    The authors declare that no competing interests exist.
  13. Csaba Pal

    Synthetic and Systems Biology Unit, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary
    For correspondence
    cpal@brc.hu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5187-9903

Funding

H2020 European Research Council (H2020-ERC-2014-CoG 648364 - Resistance Evolution)

  • Csaba Pal

Magyar Tudományos Akadémia (Lendület Programme LP 2012-32/2018)

  • Csaba Pal

Magyar Tudományos Akadémia (Postdoctoral Programme PD-007/2016)

  • Viktoria Lazar

Magyar Tudományos Akadémia (Postdoctoral Programme PD-038/2015)

  • Zoltán Farkas

National Research, Development and Innovation Office (NKFI-112294)

  • Laszlo Bodai

Magyar Tudományos Akadémia (Lendület Programme LP2009-013/2012)

  • Balazs Papp

Gazdaságfejlesztési és Innovációs Operatív Programm (GINOP-2.3.2-15-2016-00014)

  • Csaba Pal

Gazdaságfejlesztési és Innovációs Operatív Programm (GINOP-2.3.2-15-2016-00020)

  • Csaba Pal

Momentum Programme of the Hungarian Academy of Sciences (LP-2017-10/2017)

  • Csaba Pal

National Research, Development and Innovation Office (Élvonal Programme KKP 126506)

  • Csaba Pal

Wellcome Trust (WT 098016/Z/11/Z)

  • Balazs Papp

Gazdaságfejlesztési és Innovációs Operatív Programm (GINOP-2.3.2-15-2016-00026)

  • Balazs Papp

Wellcome Trust (WT 084314/Z/07/Z)

  • Csaba Pal

National Research, Development and Innovation Office (FK 128775)

  • Zoltán Farkas

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2019, Dunai et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 5,941
    views
  • 658
    downloads
  • 60
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Anett Dunai
  2. Reka Spohn
  3. Zoltán Farkas
  4. Viktoria Lazar
  5. Adam Gyorkei
  6. Gabor Apjok
  7. Gabor Boross
  8. Balazs Szappanos
  9. Gabor Grezal
  10. Aniko Farago
  11. Laszlo Bodai
  12. Balazs Papp
  13. Csaba Pal
(2019)
Rapid decline of bacterial drug-resistance in an antibiotic-free environment through phenotypic reversion
eLife 8:e47088.
https://doi.org/10.7554/eLife.47088

Share this article

https://doi.org/10.7554/eLife.47088

Further reading

    1. Evolutionary Biology
    2. Genetics and Genomics
    Julie N Chuong, Nadav Ben Nun ... David Gresham
    Research Article

    Copy number variants (CNVs) are an important source of genetic variation underlying rapid adaptation and genome evolution. Whereas point mutation rates vary with genomic location and local DNA features, the role of genome architecture in the formation and evolutionary dynamics of CNVs is poorly understood. Previously, we found the GAP1 gene in Saccharomyces cerevisiae undergoes frequent amplification and selection in glutamine-limitation. The gene is flanked by two long terminal repeats (LTRs) and proximate to an origin of DNA replication (autonomously replicating sequence, ARS), which likely promote rapid GAP1 CNV formation. To test the role of these genomic elements on CNV-mediated adaptive evolution, we evolved engineered strains lacking either the adjacent LTRs, ARS, or all elements in glutamine-limited chemostats. Using a CNV reporter system and neural network simulation-based inference (nnSBI) we quantified the formation rate and fitness effect of CNVs for each strain. Removal of local DNA elements significantly impacts the fitness effect of GAP1 CNVs and the rate of adaptation. In 177 CNV lineages, across all four strains, between 26% and 80% of all GAP1 CNVs are mediated by Origin Dependent Inverted Repeat Amplification (ODIRA) which results from template switching between the leading and lagging strand during DNA synthesis. In the absence of the local ARS, distal ones mediate CNV formation via ODIRA. In the absence of local LTRs, homologous recombination can mediate gene amplification following de novo retrotransposon events. Our study reveals that template switching during DNA replication is a prevalent source of adaptive CNVs.

    1. Developmental Biology
    2. Evolutionary Biology
    Hope M Healey, Hayden B Penn ... William A Cresko
    Research Article

    Seahorses, pipefishes, and seadragons are fishes from the family Syngnathidae that have evolved extraordinary traits including male pregnancy, elongated snouts, loss of teeth, and dermal bony armor. The developmental genetic and cellular changes that led to the evolution of these traits are largely unknown. Recent syngnathid genome assemblies revealed suggestive gene content differences and provided the opportunity for detailed genetic analyses. We created a single-cell RNA sequencing atlas of Gulf pipefish embryos to understand the developmental basis of four traits: derived head shape, toothlessness, dermal armor, and male pregnancy. We completed marker gene analyses, built genetic networks, and examined the spatial expression of select genes. We identified osteochondrogenic mesenchymal cells in the elongating face that express regulatory genes bmp4, sfrp1a, and prdm16. We found no evidence for tooth primordia cells, and we observed re-deployment of osteoblast genetic networks in developing dermal armor. Finally, we found that epidermal cells expressed nutrient processing and environmental sensing genes, potentially relevant for the brooding environment. The examined pipefish evolutionary innovations are composed of recognizable cell types, suggesting that derived features originate from changes within existing gene networks. Future work addressing syngnathid gene networks across multiple stages and species is essential for understanding how the novelties of these fish evolved.