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Genetic background of novel sequence types of CTX-M-8- and CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae from public wastewater treatment plants in São Paulo, Brazil

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Abstract

The release of extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae to the environment is a public health issue worldwide. The aim of this study was to investigate the genetic background of genes encoding ESBLs in wastewater treatment plants (WWTPs) in São Paulo, southeastern Brazil. In 2009, during a local surveillance study, seven ESBL-producing Enterobacteriaceae strains were recovered from five WWTPs and screened for ESBL genes and mobile genetic elements. Multilocus sequence typing (MLST) was carried out, and wild plasmids were transformed into electrocompetent Escherichia coli. S1-PFGE technique was used to verify the presence of high molecular weight plasmids in wild-type strains and in bla ESBL-containing E. coli transformants. Strains harbored bla CTX-M-8, bla CTX-M-15, and/or bla SHV-28. Sequencing results showed that bla CTX-M-8 and bla CTX-M-15 genes were associated with IS26. MLST revealed new sequence types for E. coli (ST4401, ST4402, ST4403, and ST4445) and Klebsiella pneumoniae (ST1574), except for one K. pneumoniae from ST307 and Enterobacter cloacae from ST131. PCR and S1-PFGE results showed CTX-M-producing E. coli transformants carried heavy plasmids sizing 48.5–209 kb, which belonged to IncI1, IncF, and IncM1 incompatibility groups. This is the first report of CTX-M-8 and SHV-28 enzymes in environmental samples, and the present results demonstrate the plasmid-mediated spread of CTX-M-encoding genes through five WWTPs in São Paulo, Brazil, suggesting WWTPs are hotspots for the transfer of ESBL genes and confirming the urgent need to improve the management of sewage in order to minimize the dissemination of resistance genes to the environment.

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Acknowledgments

We thank the team of the curators of the Institut Pasteur MLST system (Paris, France) for importing novel profiles at http://www.pasteur.fr/mlst. Nilton Lincopan is a research fellow of CNPq. We also thank Rafael Arrabaça, Flavia Nacache, and Bruna Aguiar for the essential technical support.

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Authors and Affiliations

  1. Public Health Laboratory, School of Public Health, University of São Paulo, Avenida Dr. Arnaldo 715, Cerqueira César, 01146-904, São Paulo, SP, Brazil

    Milena Dropa, Livia C. Balsalobre, Danielle E. Oliveira, Glavur R. Matté & Maria H. Matté

  2. Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, Avenida Professor Lineu Prestes, 1374, Butantã, 05508-900, São Paulo, SP, Brazil

    Nilton Lincopan, Rodrigo A. Moura & Quézia Moura da Silva

  3. Department of Clinical Analysis, School of Pharmacy, University of São Paulo, Avenida Professor Lineu Prestes, 580, Butantã, 05434-070, São Paulo, SP, Brazil

    Nilton Lincopan & Miriam Rodriguez Fernandes

  4. Environmental Company of São Paulo State (CETESB), Avenida Professor Frederico Hermann Jr, 345, Pinheiros, 05489-900, São Paulo, SP, Brazil

    Maria I. Z. Sato

Authors
  1. Milena Dropa
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  2. Nilton Lincopan
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  3. Livia C. Balsalobre
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  4. Danielle E. Oliveira
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  5. Rodrigo A. Moura
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  6. Miriam Rodriguez Fernandes
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  7. Quézia Moura da Silva
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  8. Glavur R. Matté
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  9. Maria I. Z. Sato
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  10. Maria H. Matté
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Correspondence to Milena Dropa.

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Funding

This study was funded by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), grant number 2010/12841-1.

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The authors declare that they have no competing interests.

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Responsible editor: Robert Duran

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Dropa, M., Lincopan, N., Balsalobre, L.C. et al. Genetic background of novel sequence types of CTX-M-8- and CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae from public wastewater treatment plants in São Paulo, Brazil. Environ Sci Pollut Res 23, 4953–4958 (2016). https://doi.org/10.1007/s11356-016-6079-5

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