Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content>
ABSTRACT In the present study, we provide the results of a detailed genomic analysis and the growth characteristics of a colistin-resistant KPC-3-producing Klebsiella pneumoniae sequence type 512 (ST512) isolate (the colR-KPC3-KP isolate) with a mutated pmrB and isogenic isolates of colR-KPC3-KP wit...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2019-12-01
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| Series: | mSphere |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mSphere.00551-19 |
| _version_ | 1818836624110256128 |
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| author | Cesira Giordano Adrian Klak Simona Barnini Monika A. Chlebowicz Mariacristina Menconi John W. Rossen Alexander W. Friedrich Erik Bathoorn |
| author_facet | Cesira Giordano Adrian Klak Simona Barnini Monika A. Chlebowicz Mariacristina Menconi John W. Rossen Alexander W. Friedrich Erik Bathoorn |
| author_sort | Cesira Giordano |
| collection | DOAJ |
| description | ABSTRACT In the present study, we provide the results of a detailed genomic analysis and the growth characteristics of a colistin-resistant KPC-3-producing Klebsiella pneumoniae sequence type 512 (ST512) isolate (the colR-KPC3-KP isolate) with a mutated pmrB and isogenic isolates of colR-KPC3-KP with mcr-1.2 isolated from an immunocompromised patient. From 2014 to 2017, four colR-KPC3-KP isolates were detected in rectal swab samples collected from a pediatric hematology patient at the Azienda Ospedaliero-Universitaria Pisana in Pisa, Italy. Whole-genome sequencing was performed by MiSeq sequencing (Illumina). Growth experiments were performed using different concentrations of colistin. The growth lag phases both of an isolate harboring a deletion in pmrB and of clonal variants with mcr-1.2 were assessed by the use of real-time light-scattering measurements. In the first isolate (isolate 1000-pmrBΔ, recovered in September 2014), a 17-nucleotide deletion in pmrB was detected. In subsequent isolates, the mcr-1.2 gene associated with the plasmid pIncX4-AOUP was found, while pmrB was intact. Additionally, plasmid pIncQ-AOUP, harboring aminoglycoside resistance genes, was detected. The growth curves of the first three isolates were identical without colistin exposure; however, at higher concentrations of colistin, the growth curves of the isolate with a deletion in pmrB showed longer lag phases. We observed the replacement of mutated colR-KPC3-KP pmrB by isogenic isolates with multiple resistance plasmids, including mcr-1.2-carrying pIncX4, probably due to coselection under gentamicin treatment in a patient with prolonged colR-KPC3-KP carriage. The carriage of these isolates persisted in follow-up cultures. Coselection and the advantages in growth characteristics suggest that the plasmid-mediated resistance conferred by mcr has fewer fitness costs in colR-KPC3-KP than mutations in chromosomal pmrB, contributing to the success of this highly resistant hospital-adapted epidemiological lineage. IMPORTANCE Our study shows a successful prolonged human colonization by a colistin-resistant Klebsiella pneumoniae isolate harboring mcr-1.2. An intense antibiotic therapy contributed to the maintenance of this microorganism through the acquisition of new resistance genes. The isolates carrying mcr-1.2 showed fewer fitness costs than isogenic isolates with a pmrB mutation in the chromosome. Coselection and reduced fitness costs may explain the replacement of isolates with the pmrB mutation by other isolates and the ability of the microorganism to persist despite antibiotic treatment. |
| first_indexed | 2024-12-19T03:09:34Z |
| format | Article |
| id | doaj.art-4b7162f14b10463f9b0c1b72fb88e820 |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-12-19T03:09:34Z |
| publishDate | 2019-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-4b7162f14b10463f9b0c1b72fb88e8202022-12-21T20:38:03ZengAmerican Society for MicrobiologymSphere2379-50422019-12-014610.1128/mSphere.00551-19Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content>Cesira Giordano0Adrian Klak1Simona Barnini2Monika A. Chlebowicz3Mariacristina Menconi4John W. Rossen5Alexander W. Friedrich6Erik Bathoorn7SD Ospedaliera di Microbiologia, Azienda Ospedaliero-Universitaria Pisana, Pisa, ItalyUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology, Groningen, NetherlandsSD Ospedaliera di Microbiologia, Azienda Ospedaliero-Universitaria Pisana, Pisa, ItalyUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology, Groningen, NetherlandsUO Oncoematologia Pediatrica, Azienda Ospedaliero-Universitaria Pisana, Pisa, ItalyUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology, Groningen, NetherlandsUniversity of Groningen, University Medical Center Groningen, Department of Medical Microbiology, Groningen, NetherlandsABSTRACT In the present study, we provide the results of a detailed genomic analysis and the growth characteristics of a colistin-resistant KPC-3-producing Klebsiella pneumoniae sequence type 512 (ST512) isolate (the colR-KPC3-KP isolate) with a mutated pmrB and isogenic isolates of colR-KPC3-KP with mcr-1.2 isolated from an immunocompromised patient. From 2014 to 2017, four colR-KPC3-KP isolates were detected in rectal swab samples collected from a pediatric hematology patient at the Azienda Ospedaliero-Universitaria Pisana in Pisa, Italy. Whole-genome sequencing was performed by MiSeq sequencing (Illumina). Growth experiments were performed using different concentrations of colistin. The growth lag phases both of an isolate harboring a deletion in pmrB and of clonal variants with mcr-1.2 were assessed by the use of real-time light-scattering measurements. In the first isolate (isolate 1000-pmrBΔ, recovered in September 2014), a 17-nucleotide deletion in pmrB was detected. In subsequent isolates, the mcr-1.2 gene associated with the plasmid pIncX4-AOUP was found, while pmrB was intact. Additionally, plasmid pIncQ-AOUP, harboring aminoglycoside resistance genes, was detected. The growth curves of the first three isolates were identical without colistin exposure; however, at higher concentrations of colistin, the growth curves of the isolate with a deletion in pmrB showed longer lag phases. We observed the replacement of mutated colR-KPC3-KP pmrB by isogenic isolates with multiple resistance plasmids, including mcr-1.2-carrying pIncX4, probably due to coselection under gentamicin treatment in a patient with prolonged colR-KPC3-KP carriage. The carriage of these isolates persisted in follow-up cultures. Coselection and the advantages in growth characteristics suggest that the plasmid-mediated resistance conferred by mcr has fewer fitness costs in colR-KPC3-KP than mutations in chromosomal pmrB, contributing to the success of this highly resistant hospital-adapted epidemiological lineage. IMPORTANCE Our study shows a successful prolonged human colonization by a colistin-resistant Klebsiella pneumoniae isolate harboring mcr-1.2. An intense antibiotic therapy contributed to the maintenance of this microorganism through the acquisition of new resistance genes. The isolates carrying mcr-1.2 showed fewer fitness costs than isogenic isolates with a pmrB mutation in the chromosome. Coselection and reduced fitness costs may explain the replacement of isolates with the pmrB mutation by other isolates and the ability of the microorganism to persist despite antibiotic treatment.https://journals.asm.org/doi/10.1128/mSphere.00551-19colistinfitness costmcr-1.2molecular epidemiology |
| spellingShingle | Cesira Giordano Adrian Klak Simona Barnini Monika A. Chlebowicz Mariacristina Menconi John W. Rossen Alexander W. Friedrich Erik Bathoorn Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> mSphere colistin fitness cost mcr-1.2 molecular epidemiology |
| title | Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> |
| title_full | Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> |
| title_fullStr | Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> |
| title_full_unstemmed | Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> |
| title_short | Reduced Fitness Costs of <italic toggle="yes">mcr-1.2</italic> Compared to Mutated <italic toggle="yes">pmrB</italic> in Isogenic Colistin-Resistant KPC-3-Producing <named-content content-type="genus-species">Klebsiella pneumoniae</named-content> |
| title_sort | reduced fitness costs of italic toggle yes mcr 1 2 italic compared to mutated italic toggle yes pmrb italic in isogenic colistin resistant kpc 3 producing named content content type genus species klebsiella pneumoniae named content |
| topic | colistin fitness cost mcr-1.2 molecular epidemiology |
| url | https://journals.asm.org/doi/10.1128/mSphere.00551-19 |
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