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Co-transfer of plasmid-encoded bla carbapenemases genes and mercury resistance operon in high-risk clones of Klebsiella pneumoniae

  • Genomics, Transcriptomics, Proteomics
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Abstract

Carbapenemase-producing Klebsiella pneumoniae (CP-Kp) is a real global health threat. Environmental reservoirs of resistance gene determinats, such as effluents of hospital wastewaters, are acquiring increased relevance in the selection of plasmid-encoded carbapenemase genes. The presence of Hg in environmental reservoirs may exert a positive selective pressure on tolerant bacteria, favoring the co-transfer of carbapenemase genes and mer operons. In our study, 63 CP-Kp isolates were screened for mer operons by whole genome sequencing (MySeq). Conjugation assays were performed with 24 out of 63 CP-Kp isolates harboring the mer operon. Ten transconjugants (Tc-Kp) were selected with Hg. Plasmid DNA of Tc-Kp was extracted and sequenced using single-molecule real-time (SMRT) technology (PacBio, Sequel II system) with later annotation. Plasmid analysis revealed that Tc-Kp from blaIMP-like (n = 3) showed a single plasmid belonging to IncC group with two complete mer operon next to blaIMP-like. Tc-Kp from blaVIM-1 (n = 2) harbored two plasmids, one with blaVIM-1 in an IncL, and mer operon was in an IncFIB plasmid. Tc-Kp from blaOXA-48-like (n = 5) showed 2 plasmids. blaOXA-48-like was found in an IncL plasmid, whereas mer operon was (i) in an IncR plasmid associated with blaCTX-M-15 in 3 Tc-Kp-OXA-48-like, (ii) in an IncC plasmid associated with blaCMY-2 in 1 Tc-Kp-OXA-48-like, (iii) and in an IncFIB plasmid associated with blaCTX-M-15 in 1 Tc-Kp-OXA-48-like. This is, to our knowledge, the first study to describe in K. pneumoniae producing plasmid-encoded carbapenemase, the potential impact of Hg in the co-transfer of mer operons and carbapenemase genes located in the same or different plasmids.

Graphical abstract

Key points

• Environmental reservoirs are playing an important role in the selection of carbapenemase genes.

• Conjugation assays, selecting with Hg, obtained 10 transconjugants with carbapenemase genes.

• mer operons were located in the same or different plasmids than carbapenemase genes.

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Availability of data and material

Illumina and PacBio sequence data of the 24 CP-Kp harboring plasmids of bla carbapenemase and mer operon are freely available from the NCBI BioProject database (https://www.ncbi.nlm.nih.gov/bioproject/) under accession number PRJNA738337.

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Funding

This work was supported by the Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III (project PI15-01172), and the Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía y Competitividad, the Spanish Network for Research in Infectious Diseases (REIPI RD12/0015) co-financed by European Development Regional Fund “A way to achieve Europe” ERDF.

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Authors and Affiliations

  1. Unidad de Gestión Clínica de Enfermedades Infecciosas Y Microbiología Clínica, Hospital Universitario Virgen Macarena, Seville, Spain

    Patricia Perez-Palacios, Mercedes Delgado-Valverde, Ana Gual-de-Torrella, Álvaro Pascual & Felipe Fernández-Cuenca

  2. Spanish Network for the Research in Infectious Diseases (REIPI RD16/0016, RD16/CIII/0004/0002), Instituto de Salud Carlos III, Madrid, Spain

    Patricia Perez-Palacios, Mercedes Delgado-Valverde, Ana Gual-de-Torrella, Jesús Oteo-Iglesias, Álvaro Pascual & Felipe Fernández-Cuenca

  3. Instituto de Biomedicina de Sevilla (IBIs), Hospital Universitario Virgen Macarena/CSIC/Universidad de Sevilla, Seville, Spain

    Patricia Perez-Palacios, Mercedes Delgado-Valverde, Ana Gual-de-Torrella, Álvaro Pascual & Felipe Fernández-Cuenca

  4. Laboratorio de Referencia e Investigación en Resistencia a Antibióticos e Infecciones Relacionadas con la Asistencia Sanitaria, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain

    Jesús Oteo-Iglesias

  5. Departamento de Microbiología, Universidad de Sevilla, Seville, Spain

    Álvaro Pascual

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  1. Patricia Perez-Palacios
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  2. Mercedes Delgado-Valverde
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  3. Ana Gual-de-Torrella
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  4. Jesús Oteo-Iglesias
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  6. Felipe Fernández-Cuenca
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Contributions

PPP wrote the paper, designed the study, and performed research, MDV and AGT performed the research and analyzed data, JOI contributed new methods, and AP and FFC review the manuscript and conceived and designed research.

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Correspondence to Patricia Perez-Palacios.

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Perez-Palacios, P., Delgado-Valverde, M., Gual-de-Torrella, A. et al. Co-transfer of plasmid-encoded bla carbapenemases genes and mercury resistance operon in high-risk clones of Klebsiella pneumoniae. Appl Microbiol Biotechnol 105, 9231–9242 (2021). https://doi.org/10.1007/s00253-021-11684-2

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