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Replication of the Ordered, Nonredundant Library of Pseudomonas aeruginosa strain PA14 Transposon Insertion Mutants
Published on: May 4, 2018
SPM-1-producing Pseudomonas aeruginosa ST277 clone recovered from microbiota of migratory birds
Willames M B S Martins1, Ana Clara Narciso1, Rodrigo Cayô1
1Laboratório Alerta, Division of Infectious Diseases, Department of Internal Medicine, Escola Paulista de Medicina/Universidade Federal de São Paulo-UNIFESP, São Paulo, Brazil.
Abstract:
The production of São Paulo metallo-β-lactamase (SPM-1) is the most common carbapenem resistance mechanism detected among multidrug-resistant Pseudomonas aeruginosa clinical isolates in Brazil. Dissemination of SPM-1-producing P. aeruginosa has been restricted to the nosocomial settings, with sporadic reports of environmental isolates due to contamination by hospital sewage. Herein, we described the detection and molecular characterization of SPM-1-producing P. aeruginosa recovered from the microbiota of migratory birds in Brazil. Three hundred gram-negative bacilli were recovered from cloacal and choanal swabs of Dendrocygna viduata during a surveillance study for detection of carbapenem-resistant isolates. All isolates were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. MICs were determined by agar dilution, except for polymyxin B. Antibiotic resistance genes were detected by polymerase chain reaction (PCR) followed by DNA sequencing. Transcriptional levels of oprD and efflux system encoding genes were also carried out by quantitative real-time PCR. Nine imipenem-resistant P. aeruginosa isolates were recovered with 7 of them carrying blaSPM-1. Additional resistance genes (rmtD-1, blaOXA-56,aacA4, and aac(6')-Ib-cr) were also detected in all 9 isolates. The SPM-1-producing isolates showed high MICs for all β-lactams, fluoroquinolones, and aminoglycosides, being susceptible only to polymyxin B. Interestingly, all isolates showed the same PFGE pattern and belonged to ST277. Overexpression of MexXY-OprM and MexAB-OprM was observed in those isolates that did not harbor blaSPM-1. Our results suggest that migratory birds might have played a role in the dissemination of SPM-1-producing P. aeruginosa within the Brazilian territory.
Insights
Migratory birds in Brazil carry multidrug-resistant Pseudomonas aeruginosa, a significant cause of carbapenem resistance. This finding suggests birds may contribute to spreading these dangerous bacteria within the country.
Area of Science:
- Microbiology
- Molecular Biology
- Environmental Science
Background:
- São Paulo metallo-β-lactamase (SPM-1) is a primary carbapenem resistance mechanism in multidrug-resistant Pseudomonas aeruginosa clinical isolates in Brazil.
- Previous P. aeruginosa SPM-1 dissemination was mainly linked to hospital environments and sewage contamination.
Purpose of the Study:
- To detect and molecularly characterize SPM-1-producing P. aeruginosa from migratory birds in Brazil.
- To investigate the potential role of migratory birds in the dissemination of carbapenem-resistant P. aeruginosa.
Main Methods:
- Isolation of gram-negative bacilli from cloacal and choanal swabs of migratory birds.
- Identification using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS).
- Molecular typing (PFGE, MLST), antibiotic susceptibility testing (MICs), detection of resistance genes (PCR, sequencing), and gene expression analysis (qRT-PCR).
Main Results:
- Nine imipenem-resistant P. aeruginosa isolates were recovered, with seven harboring the blaSPM-1 gene.
- All nine isolates carried additional resistance genes (rmtD-1, blaOXA-56, aacA4, aac(6')-Ib-cr) and exhibited high resistance to beta-lactams, fluoroquinolones, and aminoglycosides.
- SPM-1 producing isolates belonged to Sequence Type 277 (ST277) and showed identical PFGE patterns, suggesting clonal spread.
Conclusions:
- Migratory birds can harbor and potentially disseminate SPM-1-producing P. aeruginosa in Brazil.
- This study highlights an unexpected reservoir and potential pathway for the spread of carbapenem-resistant bacteria.
- Further surveillance is warranted to understand the ecological role of wildlife in antimicrobial resistance dissemination.

