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 beta-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: bla(CTX-M-15), bla(OXA-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: bla(CTX-M-15), bla(OXA-1), bla(TEM-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, bla(CTX-M-3) and bla(TEM-1). It probably arose by the transposition of Tn3 and ISEcp1-bla(CTX-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 bla(CTX-M-3) and bla(CTX-M-15) genes on pEK204 and pEK499/pEK516 represent separate escape events, and (iii) IncFII plasmids harboring bla(CTX-M-15) have played a crucial role in the global spread of CTX-M-15 ESBLs in E. coli.

Nursing homes as a reservoir of extended-spectrum beta-lactamase (ESBL)-producing ciprofloxacin-resistant Escherichia coli.

BACKGROUND: To assess the prevalence and risk factors for faecal carriage of fluoroquinolone-resistant, extended-spectrum beta-lactamase (ESBL)-producing, Escherichia coli (MDR E. coli) among residents in nursing homes in Northern Ireland. METHODS: Between January 2004 and May 2006, retrospective histories of hospital admissions, antimicrobial treatment and co-morbidities were collected. Faecal samples were cultured for MDR E. coli. These isolates and their ESBL genes were typed by a reference laboratory. RESULTS: Of the 294 patients included in the study, faecal samples from 119 (40.5%) grew MDR E. coli. The proportion of carriers in the different homes ranged from 0% to 75%. Epidemic strain A belonging to the ST131, O25:H4 lineage with the CTX-M-15 enzyme accounted for 58 (49%) of all isolates; its proportion varied from 0% to 100% among homes. Fifty-one percent of carriers had no history of recent hospital admission and only 13.5% had a known history of ESBL E. coli colonization or infection. In a multivariate logistic regression model, days of fluoroquinolone use [odds ratio (OR) = 1.33, 95% confidence interval (CI) 1.04-1.69, P = 0.02] and a history of urinary tract infection (OR = 2.56, 95% CI 1.37-4.78, P = 0.003) were the only variables independently associated with the risk of carrying MDR E. coli. CONCLUSIONS: The high level of faecal carriage of MDR E. coli in nursing home residents demonstrates their importance as a reservoir population. Public health measures to combat spread of these organisms should address the needs of this group.

Molecular epidemiology of multiresistant Escherichia coli isolates from community-onset urinary tract infections in Cornwall, England.

OBJECTIVES: To study the clonality of gentamicin-resistant, extended-spectrum beta-lactamase (ESBL)-negative and ESBL-producing Escherichia coli isolated from community-onset urinary tract infections (UTIs) in Cornwall. METHODS: Isolates were identified by API, susceptibilities were determined by local disc testing, and MICs were determined at the reference laboratory, both interpreted using BSAC guidelines. bla(CTX-M) genes were sought by PCR, and isolates were compared by PFGE. RESULTS: In the years 2004 and 2005, 69 E. coli were submitted by Truro (Cornwall) laboratory for reference laboratory testing: these included 14 gentamicin-resistant, ESBL-negative isolates; 45 with group 1 CTX-M enzymes; seven with group 9 CTX-M enzymes; and three with non-CTX-M ESBLs. By PFGE, nine gentamicin-resistant, ESBL-negative E. coli were distinct (<85% similarity) from all the ESBL producers, but three were related to producers of group 1 CTX-M enzymes, and two isolates were related to a non-CTX-M ESBL producer. An outbreak strain was identified, represented by 11 gentamicin-resistant and one gentamicin-susceptible isolates, all with group 1 CTX-M enzymes, and two gentamicin-resistant, ESBL-negative isolates. This was distinct by PFGE from nationally distributed CTX-M-producing strains. Five of nine patients infected with this strain had been on the same ward in a local hospital; four presented with community-onset UTIs; one inpatient developed a hospital-acquired bacteraemia. Of the other four patients presenting with community-onset UTIs, three were admitted to different hospitals and the fourth had only attended an outpatient clinic. CONCLUSIONS: Community-onset, ESBL-producing and non-producing E. coli were diverse. Two ESBL-negative isolates were closely related to a local CTX-M-producing outbreak strain, suggesting gain or loss of a bla(CTX-M)-carrying plasmid. An outbreak strain was linked with prior hospital admission and appeared not to represent genuine community acquisition.