Ana Paula D'Alincourt Carvalho-Assef

OXA-23-producing Acinetobacter baumannii: a new hotspot of diversity in Rio de Janeiro?

OBJECTIVES this study focused on the population structure of OXA-23-producing Acinetobacter baumannii clinical isolates from Rio de Janeiro, Brazil. METHODS the analysis included several genomic typing methods, including PFGE, two multilocus sequence typing (MLST) schemes, sequence group (SG) determination and bla(OXA-51-like) sequencing. The genomic context of the bla(OXA-23) gene was also evaluated using I-CeuI hybridizations and PCR assays. RESULTS congruent clustering was obtained revealing four lineages. In accordance, four new sequence types (STs) (ST131, ST132, ST133 and ST134) were obtained with the MLST-OD scheme (associated with the Oxford Database) and four (ST79, ST15 and two new allelic profiles) with the MLST-IP scheme (developed by the Institute Pasteur). Four SGs (SG1, SG4 and two new profiles) were identified, allowing the association of 70% of the isolates with European clone II. bla(OXA-51-like) sequencing revealed the presence of bla(OXA-66), bla(OXA-69), bla(OXA-95) and bla(OXA-132). CONCLUSIONS identification of new STs together with new SG profiles are findings suggestive of a local diversity hotspot that is worth exploring.

Isolation of NDM-producing Providencia rettgeri in Brazil

Laboratório de Pesquisa em Infecção Hospitalar (LAPIH), Instituto Oswaldo Cruz-FIOCRUZ, Rio de Janeiro, RJ, Brazil; Departamento de Bioquı́mica, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brazil; Fundação Estadual de Produção e Pesquisa em Saúde (FEPPS IPB-LACEN-RS), Porto Alegre, RS, Brazil; Hospital Nossa Senhora da Conceição, Porto Alegre, RS, Brazil

Detection of NDM-1-, CTX-M-15-, and qnrB4-Producing Enterobacter hormaechei Isolates in Brazil

Ana Paula D’Alincourt Carvalho-Assef, Polyana S. Pereira, Rodolpho M. Albano, Gabriela Casemiro Berião, Carolina Padilha Tavares, Thiago Pavoni Gomes Chagas, Elizabeth A. Marques, Loeci N. Timm, Renato C. F. Da Silva, Diego R. Falci, Marise D. Asensi Laboratório de Pesquisa em Infecção Hospitalar (LAPIH), Instituto Oswaldo Cruz-FIOCRUZ, Rio de Janeiro, RJ, Brazil; Departamento de Bioquímica, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brazil; Departamento de Microbiologia, Imunologia e Parasitologia, Faculdade de Ciências Médicas, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, RJ, Brazil; Fundação Estadual de Produção e Pesquisa em Saúde (FEPPS IPB-LACEN-RS), Porto Alegre, RS, Brazil; Hospital Nossa Senhora da Conceição, Porto Alegre, RS, Brazil

Escherichia coli producing KPC-2 carbapenemase: first report in Brazil☆

Carbapenem resistance in Escherichia coli isolates is rare; resistance has usually been attributed to the presence of an AmpC β-lactamase in association with the loss of porins (Poirel et al., 2004). Klebsiella pneumoniae carbapenemase (KPC)-producing E. coli have been described in a few cases, involving KPC-2 and KPC-3 acquisition in the United States (Bratu et al., 2007; Deshpande et al., 2006; Hong et al, 2005) and KPC-2 in Israel (Navon-Venezia et al., 2006) and China (Cai et al., 2008). KPC enzymes have recently been described in Brazil in Klebsiella pneumoniae (Monteiro et al., 2009; Pavez et al., 2009; Peirano et al., 2009) and Enterobacter cloacae (Zavascki et al., 2009). This study is the first reported description of the presence of KPC-2 in E. coli isolates in Brazil. Since May 2008, in the University Hospital Pedro Ernesto, a 600-bed teaching hospital located in Rio de Janeiro City, Rio de Janeiro State, has isolated a KPC-2– producing K. pneumoniae clone from different patients hospitalized in different wards (personal communication). In an effort to curb this trend, we have begun routine screening of all Enterobacteriaceae isolates with carbapenem resistance or reduced carbapenem susceptibility for presence of carbapenemases by modified Hodge test (Anderson et al., 2007). From August 2008 to May 2009, we recovered 4 carbapenemase-positive E. coli isolates from postoperative fluid collection in nephrectomy (1 isolate), urine (1 isolate), and blood (2 isolates) from patients with different clinical backgrounds hospitalized in different wards. The isolates showed polymerase chain reactionpositive results by using blaKPC primers (Peirano et al., 2009), and amplicons sequencing revealed 100% of sequence identity with blaKPC-2. The main microbiologic findings for isolates are summarized in Table 1. According to the antimicrobial susceptibility testing by disk diffusion, all isolates exhibited resistance or reduced

First Report of NDM-1-Producing Acinetobacter baumannii Sequence Type 25 in Brazil

ABSTRACT New Delhi metallo-β-lactamase 1 (NDM-1) was first identified in Brazil in Enterobacter hormaechei and Providencia rettgeri in 2013. Here, we describe the first case of NDM-1-producing Acinetobacter baumannii sequence type 25 isolated from the urinary tract of a 71-year-old man who died of multiple complications, including A. baumannii infection. The NDM-1 gene was detected by quantitative PCR, and its sequence confirmed its presence in an ∼100-kb plasmid.

Occurrence of blaOXA-23 gene in imipenem-susceptible Acinetobacter baumannii

The aim of the current study was to describe the occurrence of the blaOXA-23 gene and the ISAba1 element in imipenem-susceptible Acinetobacter baumannii strains. By performing the polymerase chain reaction mapping using combinations of ISAba1 forward primers and the blaOXA-23-like gene reverse primers, we demonstrated that the ISAba1 element did not occur upstream of the blaOXA-23 gene in five of 31 isolates, which explained the lack of resistance to imipenem despite the presence of the blaOXA-23 gene. All of the blaOXA-23-positive isolates were susceptible to imipenem and meropenem with minimal inhibitory concentration ≤ 4 µg/mL. Pulsed-field gel electrophoresis analysis revealed four genotypes among the five blaOXA-23-positive isolates. The current report of the blaOXA-23 gene in imipenem-susceptible isolates provided evidence that this gene may be silently spread in a hospital environment and highlighted the threat of undetected reservoirs of carbapenemase genes.

Clonal Dissemination of OXA-370-Producing Klebsiella pneumoniae in Rio de Janeiro, Brazil

ABSTRACT Enzymes of the OXA-48 family have become some of the most important beta-lactamases in the world. A new OXA-48 variant (OXA-370) was first described for an Enterobacter hormaechei strain isolated in Rio Grande do Sul (southern region of Brazil) in 2013. Here we report detection of the blaOXA-370 gene in 24 isolates belonging to three Enterobacteriaceae species (22 Klebsiella pneumoniae isolates, 1 Enterobacter cloacae isolate, and 1 Enterobacter aerogenes isolate) collected from five hospitals in Rio de Janeiro, Brazil, in 2013 and 2014. The isolates showed a multidrug resistance profile, and 12.5% were resistant to polymyxin B. Besides blaOXA-370, no other carbapenemase genes were observed by PCR, whereas blaOXA-1 was found in all isolates and 22 isolates (91.6%) possessed blaCTX-M-15. Molecular typing of the K. pneumoniae isolates by pulsed-field gel electrophoresis (PFGE) showed the presence of two clonal groups, i.e., KpA (21 isolates) and KpB (1 isolate). KpA was characterized as sequence type 16 (ST16) and KpB as ST1041 by multilocus sequence typing (MLST). ST16 has been observed for KPC-producing K. pneumoniae in Rio de Janeiro. Plasmid analysis performed with six representative OXA-370-producing isolates showed plasmids harboring the blaOXA-370 gene in all strains, ranging from 25 kb to 150 kb. This study suggests that there is an urgent need to investigate the presence of OXA-370 and dissemination of the K. pneumoniae ST16 clone carrying this gene in Brazil.

Detection of Carbapenemase Genes in Aquatic Environments in Rio de Janeiro, Brazil

ABSTRACT This study reveals the presence of different carbapenemase genes (blaKPC, blaNDM, blaGES, and blaOXA48-like genes) detected directly from water samples and clonal dispersion (by pulsed-field gel electrophoresis [PFGE] and multilocus sequence typing [MLST]) of KPC-2-producing Enterobacteriaceae in two important urban aquatic matrixes from Rio de Janeiro, Brazil, highlighting the role of aquatic environments as gene pools and the possibility of community spreading.

Description of genomic islands associated to the multidrug-resistant Pseudomonas aeruginosa clone ST277

Multidrug-resistant Pseudomonas aeruginosa clone ST277 is disseminated in Brazil where it is mainly associated with the presence of metallo-β-lactamase SPM-1. Furthermore, it carries the class I integron In163 and a 16S rRNA methylase rmtD that confers aminoglycoside resistance. To analyze the genetic characteristics that might be responsible for the success of this endemic clone, genomes of four P. aeruginosa strains that were isolated in distinct years and in different Brazilian states were sequenced. The strains differed regarding the presence of the genes blaSPM-1 and rmtD. Genomic comparisons that included genomes of other clones that have spread worldwide from this species were also performed. These analyses revealed a 763,863bp region in the P. aeruginosa chromosome that concentrates acquired genetic structures comprising two new genomic islands (PAGI-13 and PAGI-14), a mobile element that could be used for ST277 fingerprinting and a recently reported Integrative and Conjugative Element (ICE) associated to blaSPM-1. The genetic elements rmtD and In163 are inserted in PAGI-13 while PAGI-14 has genes encoding proteins related to type III restriction system and phages. The data reported in this study provide a basis for a clearer understanding of the genetic content of clone ST277 and illustrate the mechanisms that are responsible for the success of these endemic clones.

Emergence of the Plasmid-Mediated mcr-1 Gene in Clinical KPC-2-Producing Klebsiella pneumoniae Sequence Type 392 in Brazil

Caio Augusto Martins Aires,a Orlando Carlos da Conceição-Neto,a Thamirys Rachel Tavares e Oliveira,a Carolina Frizzera Dias,b Lara Feital Montezzi,c Renata Cristina Picão,c Rodolpho Mattos Albano,d Marise Dutra Asensi,a Ana Paula D’Alincourt Carvalho-Assefa Laboratório de Pesquisa em Infecção Hospitalar (LAPIH), Instituto Oswaldo Cruz, Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, Brazila; Comissão de Controle de Infecção Hospitalar, Hospital Santa Casa da Misericórdia de Vitória, Vitória, Brazilb; Laboratório de Investigação em Microbiologia Médica (LIMM), Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazilc; Departamento de Bioquímica, Instituto de Biologia Roberto Alcântara Gomes, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazild