Marise Dutra Asensi

First Report of KPC-2-Producing Klebsiella pneumoniae Strains in Brazil

The carbapenems are regarded as the preferential therapeutic option for treatment of serious health care-associated infections with multidrug-resistant gram-negative bacteria. Although carbapenem resistance is rarely described for the Enterobacteriaceae ([6][1]), this phenotype of resistance has

Carbapenem-hydrolysing β-lactamase KPC-2 in Klebsiella pneumoniae isolated in Rio de Janeiro, Brazil

OBJECTIVES The aim of this study was to characterize the KPC-type carbapenem-hydrolysing beta-lactamase, extended-spectrum beta-lactamases (ESBLs) and class 1 integrons among nosocomial Klebsiella pneumoniae isolated in Rio de Janeiro, Brazil. METHODS MICs were determined and isolates were screened for ESBLs, metallo-beta-lactamases (MBLs) and class A carbapenemase-producing phenotypes. The main beta-lactamases resistance genes (bla(TEM), bla(SHV), bla(CTX-M), bla(KPC), bla(IMP) and bla(VIM)) and class 1 integrons were detected by PCR followed by DNA sequencing. The genetic relatedness of isolates was determined by PFGE. RESULTS All K. pneumoniae isolates were positive for ESBL and class A carbapenemase production and negative for MBL production. All isolates were resistant to all beta-lactam antibiotics, ciprofloxacin and gentamicin, being susceptible only to tigecycline and polymyxin B. The bla(KPC-2), bla(CTX-M-1), bla(CTX-M-2), bla(CTX-M-8) and bla(SHV-11) genes were detected. PFGE analysis revealed two clonal types among KPC-producing isolates, both identified in the same hospital. CONCLUSIONS Our findings should alert medical authorities to implement stringent methods for the detection and spread control of emerging KPC-2 carbapenemases in the hospital setting in Brazil.

Dissemination of multidrug-resistant Acinetobacter baumannii genotypes carrying blaOXA-23 collected from hospitals in Rio de Janeiro, Brazil

The present study reports the dissemination of multidrug-resistant (MDR) OXA-23-producing Acinetobacter baumannii clones throughout hospitals in Rio de Janeiro, Brazil. A total of 110 imipenem-resistant A. baumannii isolates were obtained from January 2006 to September 2007 in eight hospitals. The modified Hodge test was performed to screen for carbapenemase production. Polymerase chain reaction (PCR) and DNA sequencing were performed for the detection of bla(IMP), bla(VIM), bla(OXA-23-like), bla(OXA-24-like), bla(OXA-58) and the class 1 integron. Isolates were typed by pulsed-field gel electrophoresis (PFGE) following digestion with ApaI. All the isolates were MDR and 96 (87.3%) produced the carbapenemase OXA-23. No isolates produced OXA-24, OXA-58 or the metallo-beta-lactamases IMP and VIM. The class 1 integron was absent in all isolates. The A. baumannii isolates were separated into five genotypes, with the highest prevalence of genotype A (71.8%) followed by genotype B (22.7%). Genotype A was present in seven hospitals, whilst genotype B had spread in five hospitals. The OXA-23-producing isolates belonged to all genotypes. The presence of MDR OXA-23-producing A. baumannii in different hospitals in Rio de Janeiro emphasises the need to control the use of carbapenems and to prevent the spread of these organisms in Rio de Janeiro.

Study on the bacterial midgut microbiota associated to different Brazilian populations of Lutzomyia longipalpis (Lutz & Neiva) (Diptera: Psychodidae)

The bacterial community associated with the midgut of three Brazilian Lutzomyia longipalpis (Lutz & Neiva) populations, two from endemic areas for visceral leishmaniasis (Jacobina, Bahia State and São Luís, Maranhão State) and one from a non-endemic area (Lapinha Cave, Minas Gerais State), was identified. Five groups, 35 females each, from each population were separated; a total of 175 females per collecting area were analyzed. The species identification was based on molecular and traditional bacteriological methods. All bacteria were either affiliated to non-Enterobacteriaceae, such as Acinetobacter, Burkholderia, Flavimonas, Pseudomonas and Stenotrophomonas, or and to Enterobacteriaceae, such as Citrobacter, Enterobacter, Escherichia, Klebsiella, Serratia, Pantoea, Morganella and Weeksella. Stenotrophomonas was found to be associated with all three populations studied. In addition, Serratia spp., which are well documented as laboratory contaminant of insects, were detected only in the Jacobina population. We also discuss the impact of the colonization of insect gut by bacteria on the development and transmission of pathogens.

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.

Molecular epidemiology of KPC-2- producing Klebsiella pneumoniae isolates in Brazil: the predominance of sequence type 437

The aim of this study was to investigate the genetic relatedness of 57 KPC-2-producing Klebsiella pneumoniae isolates from 5 states in Brazil, during 2006-2009. Pulse-field gel electrophoresis analysis identified 10 pulsotypes. The pulsotype designated as Kp-RJ (Klebsiella pneumoniae-Rio de Janeiro) was the dominant clone found in the states of Rio de Janeiro and Espírito Santo. Multilocus sequence typing of Kp-RJ assigned it to ST 437. Sequence types ST11, ST16, ST25, ST70, ST101, ST105, ST423, ST442, and ST443 were also identified. These results indicate the dissemination of a successful KPC-producing clone (ST437) in Brazil, which is a single locus variant of ST258.

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

Characterization of carbapenem-resistant Acinetobacter baumannii in Brazil (2008–2011): countrywide spread of OXA-23–producing clones (CC15 and CC79)

The study investigated the genetic relationship of carbapenem-resistant Acinetobacter baumannii clinical isolated from inpatients during 2008-2011 from 11 Brazilian states. Antimicrobial susceptibility profile was determined by disc diffusion method and Etest. Polymerase chain reaction was applied for carbapenemase genes, and ISAba1. Isolates were subjected to pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) for molecular typing. Most of the isolates showed high resistance rates to antibiotics tested. The blaOXA-51-like gene was found in all isolates, and 146 (94.2%) isolates were positive for blaOXA-23-like. In the most OXA-23-producing isolates, the blaOXA-23-like gene was accompanied by ISAba1. A total of 146 OXA-23-producing isolates were clustered into 28 genotypes by PFGE. Molecular analysis by MLST identified 13 sequence types (STs). The most prevalent PFGE profiles were designated as ST15 (CC15), ST1 (CC1), and ST79 (CC79). This study showed the widespread of clonal complexes of A. baumannii harboring the blaOXA-23-like gene in different Brazilian states.

Pseudomonas aeruginosa: study of antibiotic resistance and molecular typing in hospital infection cases in a neonatal intensive care unit from Rio de Janeiro City, Brazil.

This study had the objective of to analyze the demographic and bacteriologic data of 32 hospitalized newborns in an neonatal intensive care unit of a public maternity hospital in Rio de Janeiro city, Brazil, seized by Pseudomonas aeruginosa sepsis during a period ranged from July 1997 to July 1999, and to determine the antimicrobial resistance percentage, serotypes and pulsed field gel electrophoresis (PFGE) patterns of 32 strains isolated during this period. The study group presented mean age of 12.5 days, with higher prevalence of hospital infection in males (59.4%) and vaginal delivery (81.2%), than females (40.6%) and cesarean delivery (18.8%), respectively. In this group, 20 (62.5%) patients received antimicrobials before positive blood cultures presentation. A total of 87.5% of the patients were premature, 62.5% presented very low birth weight and 40.6% had asphyxia. We detected high antimicrobial resistance percentage to b-lactams, chloramphenicol, trimethoprim/sulfamethoxazole and tetracycline among the isolated strains. All isolated strains were classified as multi-drug resistant. Most strains presented serotype O11 while PFGE analysis revealed seven distinct clones with isolation predominance of a single clone (75%) isolated from July 1997 to June 1998.

Susceptibility of clinical isolates of multiresistant Pseudomonas aeruginosa to a hospital disinfectant and molecular typing

The aim of this study was to evaluate the susceptibility of 35 resistant Pseudomonas aeruginosa clinical isolates to a quaternary ammonium hospital disinfectant. The methodology was the AOAC Use-Dilution Test, with disinfectant at its use-concentration. In addition, the chromosomal DNA profile of the isolates were determined by macro-restriction pulsed field gel electrophoresis (PFGE) method aiming to verify the relatedness among them and the behavior of isolates from the same group regarding the susceptibility to the disinfectant. Seventy one percent of the isolates were multiresistant to antibiotics and 43% showed a reduced susceptibility to the disinfectant. The PFGE methodology detected 18 major clonal groups. We found isolates with reduced susceptibility to the disinfectant and we think that these are worrying data that should be further investigated including different organisms and chemical agents in order to demonstrate that microorganisms can be destroyed by biocide as necessary. We also found strains of the same clonal groups showing different susceptibility to the disinfectant. This is an interesting observation considering that only few works are available about this subject. PFGE profile seems not to be a reliable marker for resistance to disinfectants.