Conjugative IncFI plasmids carrying CTX-M-15 among Escherichia coli ESBL producing isolates at a University hospital in Germany

<p>Abstract</p> <p>Background</p> <p>Multi-drug-resistant, extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae, constitute an emerging public-health concern. Little data on the molecular epidemiology of ESBL producing <it>Escherichia coli </it>is available in Germany. Here we describe the prevalence and molecular epidemiology of ESBL producing-<it>Escherichia coli </it>isolates at a German University hospital.</p> <p>Methods</p> <p>We analysed 63 non-duplicate clinical ESBL isolates obtained over an 8-month period using PCR and sequence-based ESBL allele typing, plasmid replicon typing, phylogenetic group typing. Pulsed-field gel electrophoresis (PFGE) based genotyping and plasmid profiling was performed, as well as confirmatory DNA-based hybridization assays.</p> <p>Results</p> <p>Examination of the 63 <it>Escherichia coli </it>isolates revealed an almost equal distribution among the <it>E. coli </it>phylogenetic groups A, B1, B2 and D. High prevalence (36/63) of the CTX-M-15 gene was observed and an analysis of PFGE-based patterns revealed the presence of this CTX-M allele in multiple clones. Resistance to cefotaxime was a transferable trait and a commonly occurring 145.5 kb conjugative IncFI plasmid was detected in 65% of <it>E. coli </it>carrying the CTX-M-15 allele. The rate of transferable antibiotic resistances for GM, SXT, TET, GM-SXT-TET, SXT-TET and GM-TET was 33%, 61%, 61%, 27%, 44% and 11%, respectively. The remaining strains did not have a common IncFI plasmid but harboured transferable IncFI plasmids with sizes that ranged from 97 to 242.5 kb.</p> <p>Conclusion</p> <p>Our data demonstrate the presence of IncFI plasmids within the prevailing <it>E. coli </it>population in a hospital setting and suggest that the dissemination of CTX-M-15 allele is associated to lateral transfer of these well-adapted, conjugative IncFI plasmids among various <it>E. coli </it>genotypes.</p

Virulence factors in Escherichia coli with CTX-M-15 and other extended-spectrum -lactamases in the UK

Objectives Multiresistant Escherichia coli with CTX-M-15 extended-spectrum ß-lactamase (ESBL) are widespread in the UK. We examined their phylogenetic groups and virulence factors. Methods Clinical E. coli isolates (n = 114), collected between 2003 and 2006, were phylogenetically grouped and screened by PCR for 33 virulence factor genes. They included representatives of the five major UK epidemic E. coli strains with CTX-M-15 enzyme, as well as non-clonal isolates with CTX-M or other types of ESBLs. Results All representatives of the epidemic E. coli strains belonged to the virulent extra-intestinal phylogenetic group B2, as did 60% (34/56) of the non-clonal isolates with CTX-M-15-like enzymes and 75% (15/20) of those with non-CTX-M ESBLs. Half of those with CTX-M-9-like enzymes belonged to virulence group D. Within phylogenetic group B2, the prevalence of most virulence factors was comparable among clonal and non-clonal isolates with CTX-M enzymes, and among those with non-CTX-M ESBLs. The most frequent virulence genes were PAI, fimH, fyuA, iutA, kpsMTII, K5, traT, uidA and usp. Among the five epidemic clones, afa/draBC was specific to strain A, whereas P fimbriae were only detected in strain D, and only representatives of the B–C–E group specifically harboured sfaS, kpsMTII and K5. However, afa/draBC was also found in 30% of non-clonal isolates with CTX-M ESBLs, and no virulence gene was unique to the epidemic strains. Conclusions Most E. coli with CTX-M ESBLs belonged to virulent phylogenetic groups, mainly B2. The successful epidemic strains did not appear more virulent, but iutA and fyuA were significantly more prevalent among these than in non-clonal isolates also belonging to phylogenetic group B2. The most successful clone with CTX-M-15 enzyme (A) differed from other epidemic clones in harbouring afa/draBC, but this was also found in non-clonal isolates with CTX-M-15 enzyme

Complete nucleotide sequences of plasmids pEK204, pEK499, and pEK516, encoding CTX-M enzymes in three major Escherichia coli lineages from the United Kingdom, all belonging to the international O25:H4-ST131 clone

We determined the complete nucleotide sequences of three plasmids that encode CTX-M extended-spectrum β-lactamases (ESBLs) in pulsed-field gel electrophoresis-defined United Kingdom variants (strains A, C, and D) of the internationally prevalent Escherichia coli O25:H4-ST131 clone. Plasmid pEK499 (strain A; 117,536 bp) was a fusion of type FII and FIA replicons and harbored the following 10 antibiotic resistance genes conferring resistance to eight antibiotic classes: blaCTX-M-15, blaOXA-1, bla TEM-1, aac6′-Ib-cr, mph(A), catB4, tet(A), and the integron-borne dfrA7, aadA5, and sulI genes. pEK516 (strain D; 64,471 bp) belonged to incompatibility group IncFII and carried seven antibiotic resistance genes: blaCTX-M-15, blaOXA-1, blaTEM-1, aac6′-Ib-cr, catB4, and tet(A), all as in pEK499. It also carried aac3-IIa, conferring gentamicin resistance, and was highly related to pC15-1a, a plasmid encoding the CTX-M-15 enzyme in Canada. By contrast, pEK204 (strain C; 93,732 bp) belonged to incompatibility group IncI1 and carried only two resistance genes, blaCTX-M-3 and blaTEM-1. It probably arose by the transposition of Tn3 and ISEcp1-blaCTX-M-3 elements into a pCOLIb-P9-like plasmid. We conclude that (i) United Kingdom variants of the successful E. coli ST131 clone have acquired different plasmids encoding CTX-M ESBLs on separate occasions, (ii) the blaCTX-M-3 and bla CTX-M-15 genes on pEK204 and pEK499/pEK516 represent separate escape events, and (iii) IncFII plasmids harboring blaCTX-M-15 have played a crucial role in the global spread of CTX-M-15 ESBLs in E. coli. Copyright © 2009, American Society for Microbiology. All Rights Reserved

Molecular characterization of plasmids encoding CTX-M-15 -lactamases from Escherichia coli strains in the United Kingdom

Objectives: The UK, like other countries worldwide, has a growing problem with CTX-M ß-lactamase-producing Escherichia coli. Five major clonally related strains have been identified among CTX-M-15 producers. We characterize here the plasmids from clonal strains A and D. Methods: Plasmids were extracted and transformed into E. coli DH5a; conjugative mating was attempted on agar. MICs were determined by agar dilution. ß-Lactamases were typed by isoelectric focusing; antibiotic resistance genes and integrons were identified by PCR and sequenced. Plasmid incompatibility groups were determined by replicon PCR. Results: blaCTX-M-15 was carried by a 150 kb plasmid in strain A and a 70 kb plasmid in strain D. Conjugative transfer of cefotaxime resistance was only achieved from strain D; plasmids from both strains were transferred by transformation. The plasmid from strain A additionally carried blaTEM-1 (variably), blaOXA-1, aac(6')-Ib-cr and tet(A), as well as a class 1 integron with the gene cassettes aadA5 and dfr17; the plasmid from strain D carried blaTEM-1 consistently, also blaOXA-1, aac(6')-Ib-cr, aac3-IIa and tet(A). Both plasmids belonged to incompatibility group FII. Conclusions: blaCTX-M-15 was plasmid-mediated in both strains A and D and was linked to other antibiotic resistance genes including aac(6')-Ib-cr, which encodes an acetyltransferase, not previously found in Europe, acting on both aminoglycosides and some fluoroquinolones. Although the plasmids from the two strains differed in size, both were related and conferred similar multi-drug resistance phenotypes, suggesting that they may share a similar genetic scaffold. Both shared features with plasmids encoding CTX-M-15 ß-lactamases in E. coli from Canada and India

Malnutrition and sarcopenia are associated with increased mortality rate in nursing home residents: A prospective study

Background and aims: The aim of this study was to determine the prevalence of malnutrition risk and sarcopenia in our elderly (>= 65 years) nursing home residents and find out their association with mortality

Nationwide Survey of CTX-M-Type Extended-Spectrum β-Lactamases among Klebsiella pneumoniae Isolates in Slovenian Hospitals ▿

Among 177 extended-spectrum β-lactamase-producing Klebsiella pneumoniae isolates collected from 11 Slovenian hospitals in 2005 and 2006, 60 (34%), from eight hospitals, harbored genes for CTX-M enzymes, with blaCTX-M-15 detected by sequencing. These 60 isolates comprised 11 pulsed-field gel electrophoresis-defined strains, with several clusters of closely related isolates. Plasmids encoding CTX-M-15 enzyme were highly transmissible