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Characterisation of VIM-2-producing Pseudomonas aeruginosa isolates from lower tract respiratory infections in a Spanish hospital

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Abstract

To analyse the antimicrobial phenotype, carbapenem mechanisms, integrons, virulence factors and molecular typing of 164 Pseudomonas aeruginosa isolates recovered from lower tract respiratory samples in a Spanish hospital (1 year) as well as the patients’ clinical data. Susceptibility testing to 12 antipseudomonal agents was determined by microdilution and metallo-beta-lactamase (MBL) phenotype by double disc method. The oprD gene was studied by PCR, sequencing and comparison with P. aeruginosa PAO1 sequence. Detection and characterisation of MBLs, class 1, 2 and 3 integrons, and virulence genes were studied by PCR and sequencing. The prevalence of carbapenem-resistant P. aeruginosa (CRPA) was 26.8%. MBL phenotype was detected in 52.3% CRPA, and all of them were disseminated throughout the intensive care unit. Most of the MBL-carrying patients presented respiratory disease, mechanical ventilation, tracheostomy, bacteraemia, ≥ 30 hospitalisation days and previous treatment with carbapenems and/or ≥ 3 different antimicrobial families. The blaVIM-2 gene was the unique MBL encoding gene and was detected inside class 1 integrons. The class 1 integrons detected in 39 strains (23.8%) were associated with aminoglycosides (aadB, aadA1, aadA6, aacA4, aac(3)-I) and carbapenems resistance genes (blaVIM-2). The aac(3)-I + aadA1 and blaVIM-2 arrangements were the most prevalent ones. Thirty-one different PFGE patterns and 4 STs (ST175, ST235, ST253, ST973) were detected among the 39 intI1-positive isolates, being ST235 the most frequent. CRPA showed a great variety of alterations in oprD gene. The exoU+/exoS− genotype was detected in 82.6% of blaVIM-2-producing strains (ST235) and the exoU−/exoS+ in the remaining 17.4% (ST973).

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Acknowledgments

This publication made use of the Pseudomonas aeruginosa MLST website (https://pubmlst.org/paeruginosa/) developed by Keith Jolley and sited at the University of Oxford (Jolley and Maiden 2010, BMC Bioinformatics, 11:595). The development of this site has been funded by the Wellcome Trust.

Funding

This work was partially supported by the Departamento de Ciencia, Tecnología and Universidad from the Gobierno de Aragón, Spain (Project DGA-European Social Fund (FSE)/Grupos consolidados, B24-211130. “Investing in your future”) and by the Instituto de Salud Carlos III of Spain [projects FIS “PI12/01276”, “PI16/01381”] (Co-funded by European Regional Development Fund (FEDER) “A way to make Europe”). Alba Bellés received a SEIMC2015 mobility internship to support her work in the laboratory of Microbiología Molecular (CIBIR).

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  1. Alba Bellés

    Present address: Sección de Microbiología, Hospital Universitario Arnau de Vilanova, Lérida, Spain

Authors and Affiliations

  1. Servicio de Microbiología, IIS Aragón, Hospital Clínico Universitario Lozano Blesa, Zaragoza, Spain

    Alba Bellés, Jessica Bueno, F. Javier Castillo & Cristina Seral

  2. Área de Microbiología Molecular, Centro de Investigación Biomédica de La Rioja (CIBIR), C/Piqueras 98, 3ªplanta, 26006, Logroño, Spain

    Beatriz Rojo-Bezares, Carmen Torres & Yolanda Sáenz

  3. Área de Bioquímica y Biología Molecular, Universidad de La Rioja, Logroño, Spain

    Carmen Torres

  4. Departamento de Microbiología, Facultad de Medicina, Universidad de Zaragoza, Zaragoza, Spain

    F. Javier Castillo & Cristina Seral

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  1. Alba Bellés
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Correspondence to Yolanda Sáenz.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Informed consent was obtained from all the participants’ sampling. Given that no subject-identifiable data were generated and the surveillance activities implied no risk or burden for any individuals, the committee/hospital/authors judged that no specific ethical permission was required for individual consent.

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Bellés, A., Bueno, J., Rojo-Bezares, B. et al. Characterisation of VIM-2-producing Pseudomonas aeruginosa isolates from lower tract respiratory infections in a Spanish hospital. Eur J Clin Microbiol Infect Dis 37, 1847–1856 (2018). https://doi.org/10.1007/s10096-018-3318-3

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