Browsing by Author "Palos, Carlos"
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- Draft Genome Sequence of the First NDM-1-ProducingProvidencia stuartiiStrain Isolated in PortugalPublication . Manageiro, Vera; Sampaio, Daniel A.; Pereira, Patrícia; Rodrigues, Paulo; Vieira, Luís; Palos, Carlos; Caniça, ManuelaWe report here the draft genome sequence of the first NDM-1-producing Providencia stuartii strain isolated in Portugal. Sequence analyses revealed the presence of an incompatibility group A/C2 (IncA/C2) plasmid and of diverse acquired genes conferring resistance to β-lactams, aminoglycosides, tetracycline, macrolides, chloramphenicol, and sulfonamides. This sequence contributes to the evaluation of the spread of NDM-1 producers.
- NDM-1-producing Providencia stuartii isolates in a Portuguese HospitalPublication . Manageiro, Vera; Ferreira, Eugénia; Rodrigues, João; Sampaio, Daniel A; Vieira, Luís; Pereira, Patrícia; Rodrigues, Paulo; Palos, Carlos; Caniça, ManuelaObjective: Providencia stuartii is an opportunistic pathogen typically associated with urinary infections, and is intrinsically resistance to a wide range of antibiotics. The main aim of this study was to characterize five carbapenemase (CA) NDM-1-producing P. stuartii isolates obtained during an outbreak detected in a Hospital. Methods: MICs were obtained by the reference microdilution broth method, according to EUCAST guidelines. PCR amplification and DNA sequencing were applied to identify the presence of CA genes from class A, B and D. Direct transfer of the CA resistance phenotype was attempted by mating-out assays. Genetic relatedness was examined by PFGE. One isolate, INSRA21868, recovered from the urine of an 88-year-old male patient admitted to the intensive care unit, was selected for genetic characterization using whole-genome sequencing (WGS), performed using 150 bp paired-end reads on a MiSeq (Illumina). A set of bioinformatic web tools were used to estimate the presence of pathogenicity determinants, antibiotic resistance (AR) genes, and clinically relevant mobile genetic elements. Results: All isolates, genetically indistinguishable by PFGE, presented multidrug-resistance with non-susceptibility to all carbapenems tested. Transconjugants had AR profiles similar to those of their parental clinical isolates. All NDM-1 determinants tested were found to be carried on conjugative plasmids. In silico AR analyses using ResFinder-v2.1 revealed genes conferring resistance to β-lactams [blaNDM-1, blaCMY-4 and ΔblaDHA-1), aminoglycosides (aac(2’)-Ia, armA), tetracycline (tetB), macrolides (mphE and msrE), chloramphenicol (catB3), and sulfonamides (sul1). PlasmidFinder-v1.2 analyses revealed the presence of an IncA/C2, which has been associated with wide dissemination of blaNDM-1. In the 3’ region, the blaNDM-1 gene was adjacent to a bleomycin resistance-encoding gene (bleMBL), followed by a trpF and part of the blaDHA-1-ampR region. The ISAba125 element upstream of blaNDM-1 was interrupted by an IS26 element. Conclusion: This study emphasizes the elements involved in dissemination of nosocomial infections and the potential of WGS in epidemiological investigations in the prevention of CA dissemination among hospitals as well as to other bacterial genera.
- NDM-producing Enterobacteriaceae responsible for environmental contamination of internal medicine wards in a Portuguese HospitalPublication . Manageiro, Vera; Ferreira, Eugénia; Romão, Raquel; Rodrigues, João; Pereira, Patrícia; Cano, Manuela; Rodrigues, Paulo; Palos, Carlos; Caniça, ManuelaObjective: The hospital environment plays a role in the cross-transmission of multidrug-resistant bacteria. The aim of this study was to identify potential carbapenemase-producing Enterobacteriaceae (CPE) reservoirs that might be responsible for a NDM-1 outbreak at a hospital in Portugal. Methods: An environmental investigation was instituted in a Portuguese hospital, after the occurrence of NDM-1-producing Providencia stuartii in five patients characterized by whole-genome sequencing. CPE screening was performed in critical infection sites of internal medicine wards and intensive care units where the patients stayed during hospitalization, by using MacConkey agar supplemented with ertapenem. Air samples were also obtained for the same locations. Antimicrobial susceptibility was determined by the disk diffusion method according to EUCAST guidelines. PCR and sequencing were applied to detect and identify carbapenemase-encoding genes. PCR-based replicon typing was used to characterize the plasmids. The genetic relationship between CPE was evaluated by PFGE. Results: Sixty-three Gram-negative isolates (Acinetobacter baumannii, Burkholderia cepacia, Klebsiella pneumoniae, Providentia stuartii and Pseudomonas spp.) were obtained from the 91 surface samples collected. A total of 17 NDM-1-producers were found: 9 K. pneumoniae and 8 P. stuartii in flush handles, bathroom door handles, dinamap monitors and/or hygiene cars from internal medicine wards. No CPE were obtained from air samples. The five NDM-1-producing P. stuartii clinical strains obtained during the outbreak were all genetically indistinguishable from those collected in the hospital environment. Regarding NDM-1-producing K. pneumoniae, no clinical cases were described. However, PFGE analysis identified two clusters, one of them indistinguishable, consisting of 7 NDM-1-K. pneumoniae producers and 7 K. pneumoniae non-CPE. PCR-based replicon typing revealed the presence of the same plasmid type among NDM-1-producers, suggesting horizontal transfer within the hospital surfaces. Conclusion: These results showed the importance of the hospital environment as a potential reservoir and source of CPE. This study also emphasizes the need of early environmental screening to interrupt the transmission of CPE.