The emergence of KPC-producing isolates of K. pneumoniae in Norway and Sweden is associated with multiple import events and probable local transmission of a successful multiresistant ST258 clone, closely related to the CTX-M-15-producing ST11 clone previously described in Hungary.
BackgroundTo study the molecular characteristics of a long-term, low frequency outbreak of bla KPC-2 in a low prevalence setting involving the hospital environment.Methodology/Principal FindingsKPC-producing bacteria were screened by selective chromogenic agar and Real-Time PCR. The presence of antibiotic resistance genes was ascribed by PCRs and subsequent sequencing, and the KPC-producing isolates were phylogenetically typed using PFGE and multi-locus sequence typing. Bla KPC-2-plasmids were identified and analysed by S1-nuclease-PFGE hybridization and PCR based replicon typing. A ∼97 kb IncFII plasmid was seen to carry bla KPC-2 in all of the clinical isolates, in one of the isolates recovered from screened patients (1/136), and in the Klebsiella pneumoniae and Enterobacter asburiae isolates recovered from the environment (sinks) in one intensive care unit. The K. pneumoniae strain ST258 was identified in 6 out of 7 patients. An intergenus spread to E. asburiae and an interspecies spread to two different K. pneumoniae clones (ST27 and ST461) of the bla KPC-2 plasmid was discovered. K. pneumoniae ST258 and genetically related E. asburiae strains were found in isolates of both human and environmental origins.Conclusions/SignificanceWe document a clonal transmission of the K. pneumoniae ST258 strain, and an intergenus plasmid diffusion of the IncFII plasmid carrying bla KPC-2 in this outbreak. A major reservoir in the patient population could not be unveiled. However, the identification of a persisting environmental reservoir of strains with molecular determinants linked to human isolates, suggests a possible role of the environment in the maintenance of this long-term outbreak.
Consecutive clinical isolates of Escherichia coli (n ؍87Systemic infections with extended-spectrum--lactamase (ESBL)-producing Enterobacteriaceae are associated with severe adverse clinical outcomes (7,12,25). It is thus essential for a diagnostic microbiology laboratory to have updated methods for the detection of ESBL-producing strains, taking into account the local epidemiology of ESBL genotypes and their various expression profiles. As very little is known about ESBL genotypes in Norway, we designed a study for the detection and characterization of ESBL production in clinical isolates of Escherichia coli and Klebsiella pneumoniae with reduced susceptibilities to oxyimino-cephalosporins from routine diagnostic samples. More specifically, we examined (i) the abilities of different phenotypic methods to detect ESBL-producing strains in relation to MICs of oxyimino-cephalosporins, (ii) the molecular basis for ESBL production by typing of the most prevalent -lactamase genes (bla TEM , bla SHV , and bla CTX-M ) and the relationships between MIC profiles for oxyimino-cephalosporins and different bla groups, and (iii) the occurrence of multiple-antibiotic resistance.(The results of this study were presented in part at the European Congress of Clinical Microbiology and Infectious Diseases, Prague, Czech Republic, 2004.) MATERIALS AND METHODSStudy design. Consecutive nonduplicate isolates of E. coli and K. pneumoniae with reduced susceptibilities to oxyimino-cephalosporins (MIC Ͼ 1 mg/liter) were collected in 18 of 24 Norwegian diagnostic microbiology laboratories covering
Several studies have reported an increased incidence of candidaemia and a redistribution of species, with a decrease in the number of Candida albicans isolates. In Norway, a prospective, national surveillance study of candidaemia has been ongoing since 1991. Data from the period 1991-2003 have been published previously. The aim of this study was to follow up the incidence, species distribution and antifungal susceptibility of Candida species isolates from blood cultures in the period 2004-2012, and compare them with the corresponding findings from the period 1991-2003. Blood culture isolates of Candida species from all medical microbiological laboratories in Norway were identified and susceptibility tested at the Norwegian Mycological Reference Laboratory. A total of 1724 isolates were recovered from 1653 patients in the period 2004-2012. Comparison of the two periods showed that the average incidence of candidaemia episodes per 100 000 inhabitants increased from 2.4 (1991-2003) to 3.9 (2004-2012). The increase in incidence in the latter period was significantly higher in patients aged >40 years (p 0.001), and a marked increase was observed in patients aged >60 years (p < 0.001). In conclusion, the average incidence in Norway over a period of 22 years modestly increased from 2.4 to 3.9 per 100,000 inhabitants, this being mainly accounted for by candidaemia in the elderly. The species distribution was stable, and the rate of acquired resistance was low.
Background The clonal diversity underpinning trends in multidrug resistant Escherichia coli causing bloodstream infections remains uncertain. We aimed to determine the contribution of individual clones to resistance over time, using large-scale genomics-based molecular epidemiology.Methods This was a longitudinal, E coli population, genomic, cohort study that sampled isolates from 22 512 E coli bloodstream infections included in the Norwegian surveillance programme on resistant microbes (NORM) from 2002 to 2017. 15 of 22 laboratories were able to share their isolates, and the first 22•5% of isolates from each year were requested. We used whole genome sequencing to infer the population structure (PopPUNK), and we investigated the clade composition of the dominant multidrug resistant clonal complex (CC)131 using genetic markers previously reported for sequence type (ST)131, effective population size (BEAST), and presence of determinants of antimicrobial resistance (ARIBA, PointFinder, and ResFinder databases) over time. We compared these features between the 2002-10 and 2011-17 time periods. We also compared our results with those of a longitudinal study from the UK done between 2001 and 2011. FindingsOf the 3500 isolates requested from the participating laboratories, 3397 (97•1%) were received, of which 3254 (95•8%) were successfully sequenced and included in the analysis. A significant increase in the number of multidrug resistant CC131 isolates from 71 (5•6%) of 1277 in 2002-10 to 207 (10•5%) of 1977 in 2011-17 (p<0•0001), was the largest clonal expansion. CC131 was the most common clone in extended-spectrum β-lactamase (ESBL)-positive isolates (75 [58•6%] of 128) and fluoroquinolone non-susceptible isolates (148 [39•2%] of 378). Within CC131, clade A increased in prevalence from 2002, whereas the global multidrug resistant clade C2 was not observed until 2007. Multiple de-novo acquisitions of both bla CTX-M ESBL-encoding genes in clades A and C1 and gain of phenotypic fluoroquinolone non-susceptibility across the clade A phylogeny were observed. We estimated that exponential increases in the effective population sizes of clades A, C1, and C2 occurred in the mid-2000s, and in clade B a decade earlier. The rate of increase in the estimated effective population size of clade A (N e =3147) was nearly ten-times that of C2 (N e =345), with clade A over-represented in Norwegian CC131 isolates (75 [27•0%] of 278) compared with the UK study (8 [5•4%] of 147 isolates).Interpretation The early and sustained establishment of predominantly antimicrobial susceptible CC131 clade A isolates, relative to multidrug resistant clade C2 isolates, suggests that resistance is not necessary for clonal success. However, even in the low antibiotic use setting of Norway, resistance to important antimicrobial classes has rapidly been selected for in CC131 clade A isolates. This study shows the importance of genomic surveillance in uncovering the complex ecology underlying multidrug resistance dissemination and competition, which have impl...
Nationwide, CTX-M-producing clinical Escherichia coli isolates from the Norwegian ESBL study in 2003 (n=45) were characterized on strain and plasmid levels. Bla(CTX-M) allele typing, characterization of the genetic environment, phylogenetic groups, pulsed field gel electrophoresis (PFGE), serotyping and multilocus sequence typing were performed. Plasmid analysis included S1-nuclease-PFGE, polymerase chain reaction-based replicon typing, plasmid transfer and multidrug resistance profiling. Bla(CTX-M-15) (n=23; 51%) and bla(CTX-M-14) (n=11; 24%) were the major alleles of which 18 (78%) and 6 (55%), respectively, were linked to ISEcp1. Thirty-two isolates were of phylogenetic groups B2 and D. Isolates were of 29 different XbaI-PFGE-types including six regional clusters. Twenty-three different O:H serotypes were found, dominated by O25:H4 (n=9, 20%) and O102:H6 (n=9, 20%). Nineteen different STs were identified, where ST131 (n=9, 20%) and ST964 (n=7, 16%) were dominant. Bla(CTX-M) was found on > or =100 kb plasmids (39/45) of 10 different replicons dominated by IncFII (n=39, 87%), FIB (n=20, 44%) and FIA (n=19, 42%). Thirty-nine isolates (87%) displayed co-resistance to other classes of antibiotics. A transferable CTX-M phenotype was observed in 9/14 isolates. This study reveals that the majority of CTX-M-15-expressing strains in Norway are part of the global spread of multidrug-resistant ST131 and ST-complex 405, associated with ISEcp1 on transferrable IncFII plasmids.
This study was designed to investigate the molecular epidemiology and antibiotic-resistance characteristics of 11 carbapenem-resistant clinical isolates of Acinetobacter baumannii obtained in Norway between 2004 and 2009. Interestingly, all the isolates were linked with recent hospitalization outside Norway. The epidemiological status was investigated by multilocus sequence typing (MLST), multiplex PCR assays for major international clones, typing of bla OXA-51 -like variants and PFGE. The genotypic-resistance characteristics, including the occurrence of OXA-carbapenemase-encoding and 16S rRNA methylase-encoding genes and class 1 integrons, were investigated by PCR assays and sequencing. Seven isolates were found to harbour bla OXA-66 and belong to MLST clonal complexes (CCs) CC2 P (Pasteur Institute scheme) and CC92 B (Bartual scheme), and international clone II. One isolate harboured bla OXA-69 , and belonged to CC1 P , CC109 B and international clone I. Two isolates belonged to sequence group 9, probably a subgroup of international clone I, and one isolate belonged to sequence group 4, a proposed novel international clone. All isolates contained an acquired OXAcarbapenemase-encoding gene: bla OXA-23 -like (n59), bla OXA-24 -like (n51) and bla OXA-58 -like (n51). Four isolates with high-level aminoglycoside-resistance contained the 16S rRNA methylase-encoding armA gene. Class 1 integrons with six different variable regions were detected. Sequence analysis of gene cassettes identified four aminoglycoside (aacA4, aac(69)-Im, aadA1 and aacC1), two chloramphenicol (catB8 and cm1A5), one b-lactamase (bla and one rifampicin (arr-2) resistance gene in various combinations. In conclusion, the occurrence of A. baumannii isolates producing OXA carbapenemase and 16S rRNA methylase in Norway was related to the worldwide distribution of international clones I and II, and the emergence of novel international clones.
Our results provide support for the hypothesis that clonal transfer of cephalosporin-resistant E. coli from chicken meat to humans may occur, and may cause difficult-to-treat infections. Furthermore, these E. coli can be a source of AmpC-resistance plasmids for opportunistic pathogens in the human microbiota.
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