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Research Article

Selection of a Multidrug Resistance Plasmid by Sublethal Levels of Antibiotics and Heavy Metals

Erik Gullberg, Lisa M. Albrecht, Christoffer Karlsson, Linus Sandegren, Dan I. Andersson
Fernando Baquero, Editor
Erik Gullberg
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Lisa M. Albrecht
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Christoffer Karlsson
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Linus Sandegren
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Dan I. Andersson
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Fernando Baquero
Ramón y Cajal University Hospital
Roles: Editor
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DOI: 10.1128/mBio.01918-14
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  • FIG 1 
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    FIG 1 

    Selection coefficients as function of antibiotic concentrations during competition experiments between strains carrying the pUUH239.2 plasmid and susceptible strains at low levels of antibiotics and heavy metals. Competitions in the presence of tetracycline (A), trimethoprim (B), erythromycin (C), kanamycin (D), arsenite (E), and copper (F). Data can be found in Table S3 in the supplemental material. Standard deviations are indicated.

  • FIG 2 
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    FIG 2 

    Selection coefficients as a function of antibiotic and heavy metal concentrations during competition experiments between resistant strains carrying the pUUH239.2 plasmid and susceptible plasmid-free strains. (A) Competitions in the presence of combinations of trimethoprim and erythromycin. The total normalized concentration is the sum of the two normalized concentrations, where the concentration of each compound was first normalized to its respective MSC. Data can be found in Table S4 in the supplemental material. (B) Competitions in the presence of combinations of arsenic and tetracycline. These competitions were performed in LB medium, explaining why the selection coefficients of tetracycline in this experiment were different from the values in Fig. 1A, where Mueller-Hinton medium was used. Data can be found in Table S5. The differences in MSC with and without arsenic were significant (P value = 0.0001 in a paired t test). (C) Competitions in the presence of combinations of arsenic, tetracycline, and trimethoprim. The total normalized concentration is the sum of all normalized concentrations, where the concentration of each compound was first normalized to its respective MSC. Data can be found in Table S6. Standard deviations are indicated.

  • FIG 3 
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    FIG 3 

    Selection coefficients as function of antibiotic concentrations during competition experiments between strains carrying chromosomally carried resistance genes and susceptible strains at low levels of antibiotics. Competitions in the presence of tetracycline (A), trimethoprim (B), and erythromycin (C). Data can be found in Table S7 in the supplemental material. Standard deviations are indicated.

Tables

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  • TABLE 1 

    Strains and genotypes

    StrainGenotypeSource or reference
    DA5438E. coli MG1655 wild type (parent)Strain collection
    DA25916/pUUH239.2This study
    DA26735ΔlacIZYA::FRT galK::mTagBFP2-ampThis study
    DA26736ΔlacIZYA::FRT galK::SYFP2-ampThis study
    DA26737ΔlacIZYA::FRT galK::mTagBFP2-amp/pUUH239.2 ΔtraJ::catThis study
    DA26738ΔlacIZYA::FRT galK::SYFP2-amp/pUUH239.2 ΔtraJ::catThis study
    DA28200galK::SYFP2-FRTThis study
    DA28202galK::mTagBFP2-FRTThis study
    DA28893galK::SYFP2-FRT ΔbglGFB::FRTThis study
    DA28895galK::mTagBFP2-FRT ΔbglGFB::FRTThis study
    DA28702galK::SYFP2-FRT ΔbglGFB::dhfrThis study
    DA28704galK::mTagBFP2-FRT ΔbglGFB::dhfrThis study
    DA28706galK::SYFP2-FRT ΔbglGFB::mphThis study
    DA28708galK::mTagBFP2-FRT ΔbglGFB::mphThis study
    DA28759galK::SYFP2-FRT ΔbglGFB::tetRAThis study
    DA28761galK::mTagBFP2-FRT ΔbglGFB::tetRAThis study
  • TABLE 2 

    MICs, MSCs, and relative fitness values for the different strains useda

    StrainResistanceFitnessArseniteCu(II) sulfateTetTrmEryKan
    MSC (µM)MIC (µM)MSC (ng/ml)MIC (ng/ml)MSC (ng/ml)MIC (ng/ml)MSC (ng/ml)MIC (ng/ml)MSC (µg/ml)MIC (µg/ml)MSC (µg/ml)MIC (µg/ml)
    DA5438None (wt)1.0NA12,500NA1,300NA750NA190NA12NA0.75
    DA25916pUUH239.20.969025,00090>80,0004524,00033>32,0003960.4724
    DA28759tetRA0.99NANANANA3048,000NANANANANANA
    DA28702dhfr1.0NANANANANANA<2>32,000NANANANA
    DA28706mph1.0NANANANANANANANA<0.2>256NANA
    • ↵a Abbreviations: MSC, minimal selective concentration; Tet, tetracycline; Trm, trimethoprim; Ery, erythromycin; Kan, kanamycin; NA, not applicable; wt, wild type.

Supplemental Material

  • Figures
  • Tables
  • Additional Files
  • Table S1

    Costs of fluorescence markers. Table S1, XLSX file, 0.04 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S3

    Data for Fig. 1. Table S3, DOCX file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Figure S1

    Competitions with low initial fractions of resistant bacteria. Competition experiments at different concentrations of arsenite and different starting fractions of resistant bacteria carrying the pUUH239.2 plasmid. (A) Initial ratio of susceptible to resistant mutants, 10:1. (B) Initial ratio of susceptible to resistant mutants, 100:1. Data can be found in Table S8. Standard deviations are indicated. Download Figure S1, TIF file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S4

    Data for Fig. 2A. Table S4, DOCX file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S5

    Data for Fig. 2B. Table S5, DOCX file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S6

    Data for Fig. 2C. Table S6, DOCX file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S7

    Data for Fig. 3. Table S7, DOCX file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S9

    Summary of whole-genome sequencing data. Table S9, DOCX file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S2

    Primers used in this study. Table S2, DOCX file, 0.1 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S8

    Data for Fig. S1. Table S8, DOCX file, 0.04 MB.

    Copyright © 2014 Gullberg et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

Additional Files

  • Figures
  • Tables
  • Supplemental Material
  • Supplementary Data

    Supplementary Data

    Files in this Data Supplement:

    • Figure sf1, TIF - Figure sf1, TIF
    • Table st1, XLSX - Table st1, XLSX
    • Table st2, DOCX - Table st2, DOCX
    • Table st3, DOCX - Table st3, DOCX
    • Table st4, DOCX - Table st4, DOCX
    • Table st5, DOCX - Table st5, DOCX
    • Table st6, DOCX - Table st6, DOCX
    • Table st7, DOCX - Table st7, DOCX
    • Table st8, DOCX - Table st8, DOCX
    • Table st9, DOCX - Table st9, DOCX
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Selection of a Multidrug Resistance Plasmid by Sublethal Levels of Antibiotics and Heavy Metals
Erik Gullberg, Lisa M. Albrecht, Christoffer Karlsson, Linus Sandegren, Dan I. Andersson
mBio Oct 2014, 5 (5) e01918-14; DOI: 10.1128/mBio.01918-14

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Selection of a Multidrug Resistance Plasmid by Sublethal Levels of Antibiotics and Heavy Metals
Erik Gullberg, Lisa M. Albrecht, Christoffer Karlsson, Linus Sandegren, Dan I. Andersson
mBio Oct 2014, 5 (5) e01918-14; DOI: 10.1128/mBio.01918-14
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