Aims:  To investigate the bacterial diversity, antimicrobial resistance patterns and types of beta‐lactamase genes in Gram‐negative bacteria isolated from a hospital sewage treatment plant. Methods and Results:  Between July and December 2008, we collected samples from influent, clarifier tank effluent and chlorine contact tank effluent from a sewage treatment plant service of a hospital located in the city of Rio de Janeiro, Brazil. Of the 221 isolates identified, 40% were characterized as extended‐spectrum beta‐lactamase (ESBL) producers. Nonpathogenic micro‐organisms and some pathogenic genera were quantified. The most common ESBL‐producing isolates were Klebsiella pneumoniae, Enterobacter cloacae and Escherichia coli. The blaTEM, blaSHV and blaCTX‐M genes were detected in 82, 48 and 67% of bacterial isolates, respectively. Conclusions:  Results showed that hospital wastewater treatment plant is not suitable systems for the removal of all antibiotic‐resistant micro‐organisms present in hospital wastewaters. Significance and Impact of the Study:  This study provides evidence that bacteria resistant to multiple antibiotics and their resistance genes that are usually present in the hospital can reach the environment, even after the use of hospital wastewater treatment plants.

No 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.

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.

Carbapenem-hydrolysing b-lactamase KPC-producing isolates of Klebsiella pneumoniae are a major problem and concern. The first KPC-producing isolate was K. pneumoniae from the USA, identified in 1996, with subsequent identifications in China, Europe, Israel, Central and Brazil. 1 The KPC b-lactamases, a mostly plasmid-

These findings suggest the requirement of constant monitoring of these microorganisms in order to control the spread of these clones in the hospital environment.

A fter the first description of NDM-1 carbapenemase in a Providencia rettgeri isolate in Brazil in February 2013 (1), the public hospital where this isolate was recovered began an active surveillance in search of asymptomatic carriers and in the hospital environment. Furthermore, a retrospective study of carbapenem-resistant isolates stored at that hospital since 2012 was performed. Six NDM-1-producing Enterobacter hormaechei subsp. oharae isolates, identified by the Vitek2 system and hsp60 genotyping, were recovered and characterized by phenotypic assays and molecular techniques, such as PCR and DNA sequencing (2, 3). One isolate (CCBH10892) was recovered from a rectal swab of a patient at the intensive care unit (ICU) in September 2012 (a time period prior to the isolation of NDM-positive P. rettgeri). The others were isolated from March to May 2013 from rectal swabs of patients (n ϭ 4) and a sink (n ϭ 1) located in the same ICU.Pulsed-field gel electrophoresis (PFGE) of XbaI-digested DNA (4) showed that all isolates belonged to the same clone; the isolates were considered to be multidrug resistant, as they were susceptible only to amikacin (MIC range of 8 to 16 mg/liter) and polymyxin B (MIC Յ 1 mg/liter) by the Etest method (Fig. 1). Besides bla NDM-1 , all these isolates carried bla CTX-M-15 , qnrB4, and aac(6)-Ib genes, detected by PCR and sequencing. Plasmid analysis by restriction digests with S1 nuclease and Southern blotting (5) showed that the bla NDM-1 and qnrB4 genes were located on the same plasmids, ranging in size from 420 to 490 kb (Fig. 1).To obtain a comprehensive in-depth view of the genetic structure surrounding the bla NDM-1 , bla CTX-M-15 , and qnrB4 genes, the genomic sequence of the CCBH10892 isolate was determined on an Illumina MiSeq system. A total of 1,149,470 reads (5,373,710 bp) were assembled with Geneious assembler (Biomatters) to generate 56 contigs. We found bla NDM-1 in a 94,795-bp contig flanked by a truncated ISAba125 sequence at the right boundary and by a bleomycin resistance gene (ble MBL ) at the left (GenBank accession number KF727591). This region shared 99% identity with the NDM region present in a plasmid carried by a Klebsiella pneumoniae isolate from Taiwan (6). In this contig, some conjugation and plasmid transfer genes and a replication protein gene belonging to the IncF group were also observed. However, this replicon could not be detected by the PCR-based replicon typing (PBRT) scheme (7).The qnrB4 gene was found in a 16,569-bp contig in which ISCR1 and genes encoding permeases (sapA, sapB, and sapC) and phage shock proteins (pspA, pspB, pspC, and pspD) and AmpC bla DHA-1 (GenBank accession number KF646592) were also observed. This same region has been reported in different plasmids of other bacterial species (8).bla was integrated into the chromosome associated with an upstream ISEcp1 element in a 277,989-bp contig (part of it is in the sequence available at GenBank accession number KF727590). This transposition unit was inserted into the flhC gene, which enc...

We characterized NDM-1-producing isolates from Rio de Janeiro, Brazil. PCR was applied for resistance and virulence determinants. The genetic context of was determined by S1 nuclease pulsed-field gel electrophoresis (PFGE) and hybridization. Genotyping was performed by PFGE and multilocus sequence typing (MLST). Most isolates carried multiple resistance genes and remained susceptible to amikacin, fosfomycin-trometamol, polymyxin B, and tigecycline. The spread of NDM-1-producing was not associated with clonal expansion and appears to be associated with Tn.