Bacteremia Caused by Extended-Spectrum-β-Lactamase-Producing Escherichia coli Sequence Type ST131 and Non-ST131 Clones: Comparison of Demographic Data, Clinical Features, and Mortality

Escherichia coli producing the highly virulent, multidrug-resistant, CTX-M-15 extended-spectrum β-lactamase (ESBL), sequence type 131 (ST131), has emerged on three continents since the late 2000s. We described the molecular epidemiology, clinical features, and outcome of ESBL-producing E. coli bacteremia in Taiwan from 2005 to 2010. This study aims to determine whether the risk factors, clinical features, and outcomes of the ST131 isolate differ from those of non-ST131 isolates. From 2005 to 2010, we collected 122 nonduplicated, consecutive, ESBL-producing E. coli isolates from bloodstream infections in a 1,200-bed hospital in Taiwan. Isolates were characterized using multilocus sequence typing. Demographic data, clinical features, and outcomes were collected from medical chart records. Thirty-six (29.5%) patients with bacteremia with ESBL-producing E. coli ST131 were identified. Patients with clone ST131 were more likely to have secondary bacteremia and noncatheterized urinary tract infections (P < 0.05). Secondary bacteremia (odds ratio [OR], 5.05; 95% confidence interval [CI], 1.08 to 23.56) and urinary catheter nonuse (OR, 3.77; 95% CI, 1.17 to 12.18) were independent risk factors for the ST131 clone after adjustment. Mortality rates at day 28 were similar in ST131 and non-ST131 populations. Independent risk factors predicting mortality at day 28 included malignancy, shock, and hospital-acquired bacteremia. In ESBL-producing E. coli bloodstream infections, the ST131 clone was not associated with health-care-associated risk factors, such as urinary catheter use or antibiotic exposure. Although highly virulent and multidrug resistant, the ST131 clone was not associated with higher mortality than non-ST131 clones.

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title: Escherichia coli sequence type ST131 as the major cause of serious multidrug-resistant E. coli infections in the United States.
abstract: Escherichia coli sequence type ST131 (O25:H4), associated with the CTX-M-15 extended-spectrum beta-lactamase, has emerged internationally as a multidrug-resistant pathogen but has received little attention in the United States.



From the SENTRY and Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) surveillance programs, 127 E. coli clinical isolates from hospitalized patients across the United States in 2007, stratified by extended-spectrum cephalosporin and fluoroquinolone phenotype and bla(CTX-M-15) genotype, were assessed for phylogenetic group, ST131 status, susceptibility profile, virulence genotype, gyrA and parC sequence, and pulsed-field gel electrophoresis profile.



The 54 identified ST131 isolates (all fluoroquinolone resistant) accounted for an estimated 17% of the source populations, including 67%-69% of isolates resistant to extended-spectrum cephalosporins or fluoroquinolones, 55% of those resistant to both fluoroquinolones and trimethoprim-sulfamethoxazole, and 52% of multidrug-resistant isolates. Their distinctive virulence profiles were more extensive compared with other antimicrobial-resistant isolates but similarly extensive compared with antimicrobial-susceptible isolates. Pulsed-field profiling suggested ongoing dissemination among locales, with concentration of bla(CTX-M-15) within specific ST131 lineages. A historical ST131 isolate lacked the 2007 ST131 isolates' conserved fluoroquinolone resistance-associated single-nucleotide polymorphisms in gyrA and parC.



A single E. coli clonal group, ST131, probably caused the most significantly antimicrobial-resistant E. coli infections in the United States in 2007, thereby constituting an important new public health threat. Enhanced virulence and/or antimicrobial resistance compared with other E. coli, plus ongoing dissemination among locales, may underlie ST131's success. Urgent investigation of the sources and transmission pathways of ST131 is needed to inform mitigation efforts.

title: UK epidemic Escherichia coli strains A-E, with CTX-M-15 beta-lactamase, all belong to the international O25:H4-ST131 clone.
abstract: Uropathogenic and invasive Escherichia coli O25:H4-ST131 isolates producing CTX-M-15 extended-spectrum beta-lactamase (ESBL) enzymes have recently been shown to be disseminated across the globe. In the UK, many CTX-M-15 ESBL-producing E. coli strains have been previously defined as belonging to the epidemic strains A-E, as determined by PFGE. The present study was carried out to define the relationship between these two groups of pathogenic E. coli.



Multilocus sequence typing and PFGE were used for molecular characterization of a collection of 61 ESBL-producing E. coli isolates from across the UK.



Strains A to E all belonged to the ST131 clone, further underscoring the epidemiological importance of this lineage.



The future spread of the ST131 clone, and its UK variants, should be monitored closely and the pathogenic mechanisms explaining their success should be investigated.

title: Intercontinental emergence of Escherichia coli clone O25:H4-ST131 producing CTX-M-15.
abstract: Concomitant with the recent emergence of CTX-M-type extended-spectrum beta-lactamases (ESBLs), Escherichia coli has become the enterobacterial species most affected by ESBLs. Multiple locales are encountering CTX-M-positive E. coli, including specifically CTX-M-15. To gain insights into the mechanism underlying this phenomenon, we assessed clonality and diversity of virulence profiles within an international collection of CTX-M-15-positive E. coli.



Forty-one ESBL-positive E. coli isolates from eight countries and three continents (Europe, Asia and North America) were selected for study based on suspected clonality. Phylogenetic group, ERIC2 PCR profile, O H serotype, AmpC variant and antibiotic susceptibility were determined. Multilocus sequence typing (MLST) and PFGE provided additional discrimination. Virulence potential was inferred by detection of 46 virulence factor (VF) genes.



Thirty-six (88%) of the 41 E. coli isolates exhibited the same set of core characteristics: phylogenetic group B2, ERIC2 PCR profile 1, serotype O25:H4, AmpC EC6, ciprofloxacin resistance and MLST profile ST131. By PFGE, the 36 isolates constituted one large cluster at the 68% similarity level; this comprised 17 PFGE groups (defined at 85% similarity), some of which included strains from different countries. The 36 isolates exhibited highly (91% to 100%) similar VF profiles.



We describe a broadly disseminated, CTX-M-15-positive and virulent E. coli clonal group with highly homogeneous virulence genotypes and subgroups exhibiting highly similar PFGE profiles, suggesting recent emergence. Understanding how this clone has emerged and successfully disseminated within the hospital and community, including across national boundaries, should be a public health priority.

title: Molecular epidemiology of Escherichia coli producing CTX-M beta-lactamases: the worldwide emergence of clone ST131 O25:H4.
abstract: Since 2000, Escherichia coli producing CTX-M enzymes have emerged worldwide as important causes of community-onset urinary tract and bloodstream infections owing to extended-spectrum beta-lactamase (ESBL)-producing bacteria. Molecular epidemiological studies suggested that the sudden worldwide increase of CTX-M-15-producing E. coli was mainly due to a single clone (ST131) and that foreign travel to high-risk areas, such as the Indian subcontinent, might in part play a role in the spread of this clone across different continents. Empirical antibiotic coverage for these resistant organisms should be considered in community patients presenting with sepsis involving the urinary tract, especially if the patient recently travelled to a high-risk area. If this emerging public health threat is ignored, it is possible that the medical community may be forced, in the near future, to use carbapenems as the first choice for the empirical treatment of serious infections associated with urinary tract infections originating from the community.

title: Commonality among Fluoroquinolone-Resistant Sequence Type ST131 Extraintestinal Escherichia coli Isolates from Humans and Companion Animals in Australia
abstract: Escherichia coli sequence type 131 (ST131), an emergent multidrug-resistant extraintestinal pathogen, has spread epidemically among humans and was recently isolated from companion animals. To assess for human-companion animal commonality among ST131 isolates, 214 fluoroquinolone-resistant extraintestinal E. coli isolates (205 from humans, 9 from companion animals) from diagnostic laboratories in Australia, provisionally identified as ST131 by PCR, selectively underwent PCR-based O typing and blaCTX-M-15 detection. A subset then underwent multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE) analysis, extended virulence genotyping, antimicrobial susceptibility testing, and fluoroquinolone resistance genotyping. All isolates were O25b positive, except for two O16 isolates and one O157 isolate, which (along with six O25b-positive isolates) were confirmed by MLST to be ST131. Only 12% of isolates (25 human, 1 canine) exhibited blaCTX-M-15. PFGE analysis of 20 randomly selected human and all 9 companion animal isolates showed multiple instances of ≥94% profile similarity across host species; 12 isolates (6 human, 6 companion animal) represented pulsotype 968, the most prevalent ST131 pulsotype in North America (representing 23% of a large ST131 reference collection). Virulence gene and antimicrobial resistance profiles differed minimally, without host species specificity. The analyzed ST131 isolates also exhibited a conserved, host species-independent pattern of chromosomal fluoroquinolone resistance mutations. However, eight (89%) companion animal isolates, versus two (10%) human isolates, possessed the plasmid-borne qnrB gene (P < 0.001). This extensive across-species strain commonality, plus the similarities between Australian and non-Australian ST131 isolates, suggest that ST131 isolates are exchanged between humans and companion animals both within Australia and intercontinentally.

title: Escherichia coli O25b-ST131: a pandemic, multiresistant, community-associated strain.
abstract: Escherichia coli sequence type 131 (ST131) is a worldwide pandemic clone, causing predominantly community-onset antimicrobial-resistant infection. Its pandemic spread was identified in 2008 by utilizing multilocus sequence typing (MLST) of CTX-M-15 extended-spectrum β-lactamase-producing E. coli from three continents. Subsequent research has confirmed the worldwide prevalence of ST131 harbouring a broad range of virulence and resistance genes on a transferable plasmid. A high prevalence of the clone (∼30%-60%) has been identified amongst fluoroquinolone-resistant E. coli. In addition, it potentially harbours a variety of β-lactamase genes; most often, these include CTX-M family β-lactamases, and, less frequently, TEM, SHV and CMY β-lactamases. Our knowledge of ST131's geographical distribution is incomplete. A broad distribution has been demonstrated amongst antimicrobial-resistant E. coli from human infection in Europe (particularly the UK), North America, Canada, Japan and Korea. High rates are suggested from limited data in Asia, the Middle East and Africa. The clone has also been detected in companion animals, non-companion animals and foods. The clinical spectrum of disease described is similar to that for other E. coli, with urinary tract infection predominant. This can range from cystitis to life-threatening sepsis. Infection occurs in humans of all ages. Therapy must be tailored to the antimicrobial resistance phenotype of the infecting isolate and the site of infection. Phenotypic detection of the ST131 clone is not possible and DNA-based techniques, including MLST and PCR, are described.

title: Multidrug-resistant extraintestinal pathogenic Escherichia coli of sequence type ST131 in animals and foods.
abstract: Multidrug-resistant Escherichia coli sequence type 131 (ST131) has recently emerged as a globally distributed cause of extraintestinal infections in humans. Diverse factors have been investigated as explanations for ST131's rapid and successful dissemination, including transmission through animal contact and consumption of food, as suggested by the detection of ST131 in a number of nonhuman species. For example, ST131 has recently been identified as a cause of clinical infection in companion animals and poultry, and both host groups have been confirmed as faecal carriers of ST131. Moreover, a high degree of similarity has been shown among certain ST131 isolates from humans, companion animals, and poultry based on resistance characteristics and genomic background and human and companion animal ST131 isolates tend to exhibit similar virulence genotypes. However, most ST131 isolates from poultry appear to possess specific virulence genes that are typically absent from human and companion animal isolates, including genes associated with avian pathogenic E. coli. Since the number of reported animal and food-associated ST131 isolates is quite small, the role of nonhuman host species in the emergence, dissemination, and transmission of ST131 to humans remains unclear. Nevertheless, given the profound public health importance of the emergent ST131 clonal group, even the limited available evidence indicates a pressing need for further careful study of this significant question.

title: Absence of CTX-M Enzymes but High Prevalence of Clones, Including Clone ST131, among Fecal Escherichia coli Isolates from Healthy Subjects Living in the Area of Paris, France
abstract: Quinolone-resistant and CTX-M-15-producing Escherichia coli isolates belonging to clone ST131 have been reported in the community. This study was designed to identify these E. coli isolates in the stools of 332 independent healthy subjects living in the area of Paris, France. Stools were plated on media without antibiotics, in order to obtain the dominant (Dm) fecal E. coli strain, and with nalidixic acid (NAL) and cefotaxime. Quinolone susceptibility, phylogenetic groups, and molecular profiles, including multilocus sequence types (ST), were determined for all NAL-resistant (NAL-R) isolates. Groups were also determined for the Dm strains from participants with NAL-R isolates and from a subgroup without NAL-R isolates. All B2 isolates were typed; pulsed-field gel electrophoresis was performed for the ST131 isolates, and the results were compared with those for intercontinental clone ST131. Two participants (0.6%) had extended-spectrum β-lactamase-producing (SHV-2, TEM-52) fecal E. coli isolates, and 51 (15%) had NAL-R isolates; 51% of NAL-R isolates belonged to phylogenetic group A, 31% to group D, 16% to group B2, and 2% to group B1. The Dm strain was NAL-R in 3.3% of the 332 subjects. Forty-nine percent of the NAL-R isolates belonged to clones: ST10 and ST606 for group A isolates, ST117 and ST393 for group D isolates. Of all B2 isolates studied from 100 subjects (8 NAL-R strains; 19 NAL-susceptible dominant strains), 52% belonged to three clones: ST131 (n = 7), ST95 (n = 4), and ST141 (n = 3). This is the first study to show the presence of fecal E. coli isolates of clone ST131 in 7% of independent healthy subjects not colonized by CTX-M-15-producing isolates.

title: Transmission of an Extended-Spectrum-Beta-Lactamase-Producing Escherichia coli (Sequence Type ST131) Strain between a Father and Daughter Resulting in Septic Shock and Emphysematous Pyelonephritis
abstract: Probable transmission of an extended-spectrum-beta-lactamase-producing Escherichia coli strain (sequence type ST131) between a father and daughter was documented. The father developed severe, recurrent pyelonephritis with multiple small abscesses; the daughter later developed septic shock, bacteremia, and extensive emphysematous pyelonephritis. This multidrug-resistant E. coli clone appears to be highly pathogenic and transmissible.

title: Community Transmission in the United States of a CTX-M-15-Producing Sequence Type ST131 Escherichia coli Strain Resulting in Death
abstract: A middle-aged woman developed fatal urosepsis due to a multidrug-resistant Escherichia coli strain representing sequence type ST131, a recently emerged, disseminated, multidrug-resistant extraintestinal pathogen, after presumably having acquired it from her extensively antibiotic-exposed sister with chronic recurrent cystitis. Susceptibility results (reported on day 4) showed resistance to the initially selected regimen.

title: Escherichia coli Pyomyositis: an emerging infectious disease among patients with hematologic malignancies.
abstract: Pyomyositis is typically caused by gram-positive bacteria, especially Staphylococcus aureus. Few cases of Escherichia coli pyomyositis have been reported, including only 1 involving a patient with a hematologic malignancy.



The clinical microbiology database at The M. D. Anderson Cancer Center (Houston, TX) was reviewed for the period January 2003 through December 2007 to identify cases of E. coli pyomyositis. Clinical characteristics, laboratory and radiologic findings, treatment, and outcomes were recorded. Available isolates underwent phylogenetic group determination, virulence genotyping, multilocus sequence typing, repetitive-element polymerase chain reaction, and pulsed-field gel electrophoresis.



Six cases of E. coli pyomyositis were identified. All patients were receiving chemotherapy for a hematologic malignancy; 5 were severely neutropenic. Three patients became hypotensive, 2 required intensive care, and 2 (33%) died, despite receiving carbapenem therapy. All E. coli isolates were fluoroquinolone resistant; 55% produced an extended-spectrum beta-lactamase (ESBL). Five of 6 available isolates belonged to phylogenetic group B2, had similar virulence factor profiles, exhibited > 95% similar repetitive-element polymerase chain reaction profiles, and represented sequence type ST131; however, all had unique pulsed-field gel electrophoresis profiles.



E. coli pyomyositis has emerged as a serious problem among our patients with hematologic malignancy. It usually is caused by members of E. coli ST131, a recently identified cause of fluoroquinolone-resistant, ESBL-positive E. coli infection worldwide. Awareness of this emerging syndrome and the usual causative agent is important to ensure appropriate management when febrile, neutropenic patients with hematologic malignancy exhibit signs of localized muscle infection.

title: Clinical and Molecular Characteristics of Extended-Spectrum- β-Lactamase-Producing Escherichia coli Causing Bacteremia in the Rotterdam Area, Netherlands
abstract: We investigated the clinical and molecular characteristics of bacteremia caused by extended-spectrum-β-lactamase (ESBL)-producing Escherichia coli over a 2-year period (2008 to 2009) in the Rotterdam region (including 1 teaching hospital and 2 community hospitals) of Netherlands. The majority of patients presented with community onset urinary and intra-abdominal infections, with an increase in prevalence during 2009. The majority of E. coli isolates produced CTX-M-15, and 4 sequence types (ST38, ST131, ST405, and ST648) predominated. There were significant differences in clinical and molecular characteristics between the 2 community hospitals.

title: Molecular Characteristics of Extended-Spectrum-β-Lactamase-Producing Escherichia coli Isolates Causing Bacteremia in the Calgary Health Region from 2000 to 2007: Emergence of Clone ST131 as a Cause of Community-Acquired Infections
abstract: A study was designed to characterize extended-spectrum-β-lactamase (ESBL)-producing Escherichia coli isolates causing bacteremia over an 8-year period (2000 to 2007) in a large well-defined geographical region. Molecular characterization was done by using isoelectric focusing; PCR; and sequencing of the blaCTX-M-, blaTEM-, blaOXA-, blaSHV-, and plasmid-mediated quinolone resistance determinants. Genetic relatedness was determined by pulsed-field electrophoresis with XbaI and multilocus sequence typing. A total of 67 patients with incident bloodstream infections were identified, and the majority presented with community-acquired infections involving the urinary and biliary tracts. Of the 67 ESBL-producing E. coli isolates recovered, 60 (90%) were positive for blaCTX-M genes; 32 (48%) produced CTX-M-15, 25 (37%) produced CTX-M-14, 1 (2%) produced CTX-M-24, 1 (2%) produced CTX-M-2, and 1 (2%) produced CTX-M-3, while 2 (3%) produced TEM-52 and 5 (7%) produced SHV-2. Twenty-four (36%) isolates were positive for aac(6′)-Ib-cr. The majority of isolates were resistant to ciprofloxacin (60 [90%] isolates) and gentamicin (40 [60%] isolates). The occurrence of ESBL-producing isolates was stable during the first 5 years, but there was a substantial increase from 2005 to 2007, mostly due to clone ST131, which produces CTX-M-15 and CTX-M-14, in blood cultures submitted from the community. Our results illustrated that E. coli clone ST131, which coproduces CTX-M-15, OXA-1, TEM-1, and aac(6′)-Ib-cr, has emerged as an important cause of community-onset bacteremia caused by ESBL-producing E. coli isolates; and this is the first study to identify CTX-M-14 in E. coli clone ST131.

title: Analysis of a Uropathogenic Escherichia coli Clonal Group by Multilocus Sequence Typing
abstract: Although many strain typing methods exist for pathogenic Escherichia coli, most have drawbacks in terms of resolving power, interpretability, or scalability. For this reason, multilocus sequence typing (MLST) is an appealing alternative. However, its applicability to different pathogens in specific epidemiologic contexts is not well understood. Here, we applied a previously established MLST method based on housekeeping genes to a well-characterized collection of uropathogenic E. coli isolates to compare the discriminatory ability of this procedure with that of enterobacterial repeat intergenic consensus (ERIC2) PCR, serogrouping, and pulsed-field gel electrophoresis (PFGE). Among 45 E. coli isolates studied, 17 different multilocus sequence types (ST) were identified. One MLST group (designated ST69 complex) was comprised of 22 isolates, all belonging to uropathogenic and bacteremic E. coli strains previously defined as clonal group A (CgA) by ERIC2 PCR. The ST69 strains contained five different serogroups and 14 PFGE types. ERIC2 PCR CgA strains belonging to different MLST groups were also identified. Interestingly, one cow E. coli isolate, previously shown by PFGE to be closely related to a human uropathogenic CgA strain, was found to cluster with the ST69 strains. All of the other animal and environmental CgA isolates had different MLST profiles. The discriminatory power of this MLST method based on housekeeping genes appears to be higher than that of ERIC2 PCR but lower than that of PFGE for epidemiologic study of uropathogenic E. coli.

title: Epidemiology of urinary tract infections caused by extended-spectrum beta-lactamase-producing Escherichia coli.
abstract: To describe the epidemiology of urinary tract infections caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli.



We performed three studies: a trend study (1999 to 2004) to assess the evolution and origin of ESBL-producing E. coli isolates from urine samples; a comparison of the susceptibility patterns of ESBL-producing E. coli and a random sample of non-ESBL-producing E. coli isolated in the same interval, and a retrospective chart review to determine the risk factors for acquisition of, and outcomes from, urinary tract infections caused by ESBL-producing E. coli (n = 61) compared with a random sample of non-ESBL-producing E. coli urinary tract infections (n = 61) from patients attending our institution and matched by temporal occurrence.



ESBL-producing E. coli significantly increased from 2 (0.20%) to 89 (5.52%) isolates per year (P value for trend = 0.000). Of the 161 patients with urinary tract infections caused by ESBL-producing E. coli, 100 (62%) were attending ambulatory health centers, and 61 (38%) were attending the hospital. ESBL-producing E. coli showed a significant reduction in the susceptibility to most antimicrobials, although carbapenems and fosfomycin retained significant activity. The chart review study showed that previous treatment with fluoroquinolones (odds ratio 12.98, 95% confidence interval 1.81 to 106.51, P = 0.017) and the presence of a urinary catheter (odds ratio 2.64, 95% confidence interval 1.01 to 6.88, P = 0.047) were independent risk factors associated with infections caused by ESBL-producing E. coli.



ESBL-producing E. coli is a problem of increasing importance. Our study results may help physicians select appropriate antimicrobial therapy in patients suspected of having urinary tract infections caused by ESBL-producing E. coli.

title: Emergence of clonal groups O1:HNM-D-ST59, O15:H1-D-ST393, O20:H34/HNM-D-ST354, O25b:H4-B2-ST131 and ONT:H21,42-B1-ST101 among CTX-M-14-producing Escherichia coli clinical isolates in Galicia, northwest Spain.
abstract: CTX-M enzymes, mainly CTX-M-14 and CTX-M-15, have emerged as the most prevalent extended-spectrum β-lactamase (ESBL) type produced by Escherichia coli in Spain, with successful dissemination of clonal group O25b:H4-B2-ST131 producing CTX-M-15 within the hospital and community settings. However, until now CTX-M-14-producing E. coli in Spain had been shown to belong to a wide variety of serotypes with no predominance of a certain clonal group. In the present study, 654 E. coli strains positive for ESBL production obtained between 2005 and 2008 from inpatients and outpatients of four hospitals in Galicia, northwest Spain, were analysed. The strains were characterised with regard to ESBL enzymes, serotype, virulence genes, phylogenetic group, multilocus sequence type, and pulsed-field gel electrophoresis of XbaI-digested DNA. As a result, the emergence of certain clonal groups of extraintestinal pathogenic E. coli producing CTX-M-14 has been detected in this geographic area, including O1:HNM-D-ST59, O15:H1-D-ST393/ST1394, O20:H34/HNM-D-ST354, O25b:H4-B2-ST131 and ONT:H21,42-B1-ST101. These five clonal groups showed a high virulence potential as they harboured more than eight virulence factors, which could explain their successful dissemination.