Doroti de Oliveira Garcia

Coproduction of Novel 16S rRNA Methylase RmtD and Metallo-β-Lactamase SPM-1 in a Panresistant Pseudomonas aeruginosa Isolate from Brazil

ABSTRACT Serious infections with Pseudomonas aeruginosa are frequently treated with the combination of a β-lactam antimicrobial and an aminoglycoside. P. aeruginosa strain PA0905 was isolated in 2005 from an inpatient in Brazil. It showed a panresistant phenotype that included resistance to β-lactams, aminoglycosides, and fluoroquinolones. The β-lactam resistance was conferred by the production of the metallo-β-lactamase SPM-1. No inhibitory zone was observed when a disk diffusion test was performed with the semisynthetic aminoglycoside arbekacin, raising suspicion of 16S rRNA methylase production. A cloning experiment subsequently revealed the presence of a novel 16S rRNA methylase, RmtD, which accounted for the high-level resistance to all 4,6-disubstituted deoxystreptamine aminoglycosides, such as amikacin, tobramycin, and gentamicin. RmtD shared a moderate degree of identity with RmtA, another 16S rRNA methylase that was initially reported to occur in P. aeruginosa in Japan in 2003. This is the first identification of aminoglycoside resistance mediated by a 16S rRNA methylase in South America. This is also the first report to document coproduction of a metallo-β-lactamase and a 16S rRNA methylase, a combination that would severely compromise therapeutic options for the infected patients.

Multiclonal Outbreak of Klebsiella pneumoniae Producing Extended-Spectrum β-Lactamase CTX-M-2 and Novel Variant CTX-M-59 in a Neonatal Intensive Care Unit in Brazil

ABSTRACT An outbreak of cephalosporin-resistant Klebsiella pneumoniae occurred in a neonatal intensive care unit in São Paulo, Brazil. Of the 10 pulsotypes identified during the outbreak and follow-up periods, nine produced CTX-M-2 or its new variant CTX-M-59 and one produced SHV-5. blaCTX-M-2/59 genes were located on closely related plasmids that were transferable.

High Prevalence of Metallo-β-Lactamase and 16S rRNA Methylase Coproduction among Imipenem-Resistant Pseudomonas aeruginosa Isolates in Brazil

ABSTRACT Rates of metallo-β-lactamase and 16S rRNA methylase production were investigated in 51 imipenem-resistant Pseudomonas aeruginosa clinical isolates collected from hospitals in São Paulo, Brazil. Of them, 57% and 75% produced SPM-1 and RmtD, respectively. Of note, 51% produced both enzymes, suggesting that their coproduction is already common in this geographic area.

Silent dissemination of colistin-resistant Escherichia coli in South America could contribute to the global spread of the mcr-1 gene.

During a Brazilian multicentric antimicrobial resistance surveillance study, colistin resistance was investigated in 4,620 Enterobacteriaceae isolated from human, animal, food and environmental samples collected from 2000 to 2016. We present evidence that mcr-1-positive Escherichia coli has been emerging in South America since at least 2012, supporting a previous report on the possible acquisition of mcr-1-harbouring E. coli by European travellers visiting Latin American countries.

Coproduction of 16S rRNA Methyltransferase RmtD or RmtG with KPC-2 and CTX-M Group Extended-Spectrum β-Lactamases in Klebsiella pneumoniae

ABSTRACT Eight Klebsiella pneumoniae clinical strains with high-level aminoglycoside resistance were collected from eight hospitals in São Paulo State, Brazil, in 2010 and 2011. Three of them produced an RmtD group 16S rRNA methyltransferase, RmtD1 or RmtD2. Five strains were found to produce a novel 16S rRNA methyltransferase, designated RmtG, which shared 57 to 58% amino acid identity with RmtD1 and RmtD2. Seven strains coproduced KPC-2 with or without various CTX-M group extended-spectrum β-lactamases, while the remaining strain coproduced CTX-M-2.

Population Structure Analysis of Carbapenem-Resistant Acinetobacter baumannii Clinical Isolates from Brazil Reveals Predominance of Clonal Complexes 1, 15, and 79

ABSTRACT The population structure of 71 carbapenem-resistant Acinetobacter baumannii clinical isolates from several hospitals in Brazil was investigated by ApaI pulsed-field gel electrophoresis, blaOXA-51-like subtyping, and multilocus sequence typing (Institute Pasteur scheme). In addition to the predominance of strains carrying blaOXA-23, we detected the presence of blaOXA-72 and blaOXA-231. We observed a predominance of clonal complex 1 (CC1), CC15, and CC79 and representative strains of the worldwide-disseminated international clone I.

Nosocomial infections caused by Elizabethkingia meningoseptica: an emergent pathogen

We hereby describe the clinical and epidemiological features and, outcomes of nine patients with Elizabethkingia meningoseptica infections in two hospitals over a 2-year period. All infections caused by this pathogen were nosocomial, or healthcare associated infections, in hemodialysis settings whereas none was correlated with hospital outbreaks.

Outbreak of carbapenem-resistant Klebsiella pneumoniae: two-year epidemiologic follow-up in a tertiary hospital

This study describes a carbapenem-resistant Klebsiella pneumoniae (CRKP) outbreak that occurred from October 2008-December 2010. Polymerase chain reaction assays were performed to detect the blaKPC gene and molecular typing was performed using pulsed-field gel electrophoresis (PFGE). There were 33 CRKP infections; PFGE revealed five genotypes: genotype A in five (15%), B in 18 (55%), C in eight (24%) and two unique profiles. Genotype B was disseminated in all hospital units and belonged to the same clone identified in 11 different hospitals in the state of São Paulo. Sixteen (48%) patients died. Seven isolates (21%) were resistant to polymyxin B and 45% were resistant to tigecycline and amikacin.

First report of investigation into an outbreak due to carbapenemase-producing Klebsiella pneumoniae in a tertiary Brazilian hospital, with extension to a patient in the community

This is the description of an investigation into an outbreak of carbapenemase-producing Klebsiella pneumoniae (KPC) that involved 22 inpatients in a public Brazilian hospital specialising in cardiac surgery, where these bacteria had never been isolated before. The dissemination of KPC also involved an external patient. This is the first description of a KPC outbreak in a Brazilian hospital.

First report of a NDM-producing Providencia rettgeri strain in the state of São Paulo

Carbapenemases are enzymes known to hydrolase almost all types of -lactam antibiotics and their spread among bacteria of clinical relevance are one of the main global challenges to be overcome in the next years. NDM stands for New Delhi Metallo-lactamase which was firstly identified in 2009 in a Sweden patient who had been previously hospitalized in New Delhi, India.1 Although other types of metallo-lactamases are currently present in Brazil (until now SPM-1 has been the most prevalent in clinical isolates2), the first occurrence of NDM producer was reported in a clinical isolate of Providencia rettgeri in the beginning of 2013 in Porto Alegre/RS, Southern region of Brazil.3 Since then, other cases have been reported in Brazil. A 55 year-old diabetic and hypertensive man was admitted to a tertiary public hospital (Hospital Heliópolis) in the São Paulo city to undergo amputation of the 4th toe due osteomyelitis complication. The patient had a history of gastric surgery for peptic ulcer five years before, but no report of hospitalization or travel in the past 12 months. Four months earlier, the man reported a lesion in the same toe caused by a trauma with iron sheet and high temperature solder. One month after the initial trauma the patient presented worsening of pain and swelling of 4th right toe and was at that time admitted to a tertiary public hospital from city of São Paulo (Hospital Vila Alpina) where he received intravenous ciprofloxacin and clindamycin therapy to treat the bone infection (400 mg q12h and 600 mg q6h × 14d, respectively) without success. During hospitalization at Hospital Heliópolis for amputation of the 4th toe the patient received intravenous ceftriaxone and clindamycin therapy (1 g q12h and 600 mg q6h × 3d, respectively) and then was discharged with no signs of infection to be followed up in the outpatient service. Results from bone culture of the amputated toe performed at Hospital Heliópolis revealed the presence of Gram-negative bacteria. The susceptibility tests showed a carbapenem resistance profile: Etest positive for imipenem (MIC >32 mg/L), meropenem (MIC >32 mg/L) (bioMérieux Clinical Diagnostics, France, according to CLSI 2015 breakpoints4) with Modified Hodge Test negative, and positive for metallo-lactamase (M L) production (Carbapenembac-Metalo® kit). The bacterium was identified as Providencia rettgeri, according MicroScan system (Beckman Coulter, Inc.), sensitive to amikacin (MIC ≤16 mg/L), gentamicin (MIC ≤4 mg/L), and tobramycin (MIC ≤4 mg/L). In order to identify the type of M L produced by the clinical isolate and to confirm the previous results the strain was sent to a reference laboratory at Instituto Adolf Lutz – São Paulo. The presence of blaNDM gene by Multiplex (KPC, NDM and OXA-48) and Simplex (NDM) PCR and the species identity was confirmed by Vitek-MS system (bioMérieux Clinical Diagnostics, France). The clinical isolate is under investigation in order to characterize molecular aspects of blaNDM gene and its origin. To the best of our knowledge, this is the first official report of a NDM producer Enterobacteriaceae isolated in the state of São Paulo. Worldwide dissemination of broad-spectrum carbapenemases is a real threat for the modern medicine. New Delhi Metallo-lactamase (NDM) is a new type of MetalloLactamase that precludes the use of last resort carbapenems. In Brazil, the occurrence of NDM is emerging, especially in the South and Southeast regions. In conclusion, the first NDM producer strain has been detected in the most populous state of Brazil with the largest economy, underscoring the need to urgently implement even more efforts to identify and prevent the spreading of this threat in Brazil.