Uropathogenic isolates of Escherichia coli (UPEC) contain blocks of DNA, termed pathogenicity islands (PAIs), that contribute to their virulence. Two multiplex PCR assays were developed to detect eight PAI markers among 50 commensal E. coli and 100 UPEC isolates. In total, 40% of commensal isolates and 93% of UPEC carried PAIs. Despite this difference, the distribution of various PAIs showed the same pattern in both groups, with the most prevalent being PAI IV(536) (38% commensal vs. 89% UPEC), followed by PAI I(CFT073) (26% vs. 73%), PAI II(CFT073) (14% vs. 46%), PAI II(J96) (8% vs. 34%), PAI I(536) (8% vs. 33%) and PAI II(536) (4% vs. 20%). PAI III(536) was detected only in UPEC (2%), while PAI I(J96) was not detected in any isolate. Although the mean number of PAIs per isolate was higher among UPEC (2.97) than in commensal (0.98) isolates, there were no statistical differences among group B2 E. coli from the two origins; however, commensal isolates from groups D and B1 appeared to be less virulent than pathogenic isolates. Regardless of their phylogenetic group, nearly all the commensal and UPEC isolates with the same number of PAIs had the same PAI combinations. Although group B2 E. coli are uncommon among commensal intestinal flora, they are highly virulent when present, suggesting that the intestinal flora may act as a reservoir for bacteria that can cause urinary tract infection.

E. coli isolates resistant to quinolones, trimethoprim/sulfamethoxazole and especially fluoroquinolones were associated with reductions in virulence traits and shifts to non-B2 phylogenetic groups. Moreover, fluoroquinolone resistance usually occurred in low-virulence E. coli group A isolates rather than in isolates from groups B2 and D which had lost virulence traits. CGA accounted for 23% of trimethoprim/sulfamethoxazole-resistant E. coli producing pyelonephritis.

A new CTX-M-type ␤-lactamase (CTX-M-9) has been cloned from a clinical cefotaxime-resistant Escherichia coli strain. Despite the close identity that exists between the CTX-M-9 and Toho-2 ␤-lactamases (88%), the 35 amino acids located between residues Ala-185 and Ala-219 are totally different in both enzymes. Outside of this region there are only six amino acids substitutions between both proteins.Not long after the beginning of the use of the extendedspectrum ␤-lactam antibiotics, extended-spectrum ␤-lactamases (ESBLs) were detected in Europe and the United States and have now become a serious problem around the world. ESBLs are most often derivatives of TEM or SHV enzymes. However, there is a small growing family of plasmid-mediated ESBLs of Ambler class A that are not closely related to TEM or SHV ␤-lactamases but that show homology to chromosomal ␤-lactamases of Klebsiella oxytoca, including CTX-M-1 (MEN-1) (3, 5-7), CTX-M-2 (4, 6), CTX-M-3 (14), CTX-M-4 (12, 13), 20), Toho-1 (15), Toho-2 (17), and two different enzymes, both designated CTX-M-5, which will be referred to here as CTX-M-5 (8) and 20). In this report we present a new ␤-lactamase closely related to the ␤-lactamases in this family.In 1996, a cefotaxime-resistant Escherichia coli strain (785-D) against which synergy of cefotaxime with clavulanic acid was found was detected by a conventional disk diffusion susceptibility test. The strain was isolated from the urine of a 65-year-old woman who had a urinary tract infection and diabetes mellitus type 2. Two months before the isolation, the patient underwent a nephroureterectomy because of a renal carcinoma.The MICs of the ␤-lactam antibiotics were determined by the Etest (Biodisk, Solna, Sweden). The ␤-lactamase crude cell extracts were prepared from 250-ml cultures in Luria-Bertani (LB) broth (Oxoid, Basingstoke, United Kingdom). Washed, centrifuged cell pellets were subjected to three cycles of 15-s sonication treatments at 4°C, and the supernatants of the sonic extracts were frozen at Ϫ20°C until they were tested. ␤-Lactamases were characterized initially by isoelectric focusing as described previously (2) in polyacrylamide gels with a pH gradient from 4 to 11 (SERVALYT 4-9 T, 9-11 T; Serva, Heidelberg, Germany). Enzyme activities in the gel were detected by the iodometric method (2).Substrate hydrolysis in sonic extracts was monitored spectrophotometrically with a Biochrom 4060 spectrophotometer (Pharmacia, Uppsala, Sweden) as described previously (19).Conjugation studies were performed in a solid medium as described previously (19) by using E. coli 785-D (susceptible to kanamycin) and E. coli HB101 (Nal r Kan r ) as the donor and the recipient, respectively. Kanamycin (50 g/ml) and cefotaxime (4 g/ml) were used for transconjugant selection. One of the transconjugants (E. coli MSP492) was used for further experiments.Extraction of plasmid DNA was by the alkaline lysis procedure reported previously (18). Cloning of the cefotaxime resistance gene was as follows: plasmid DNA from E. coli MSP492 was pa...

The prevalence of qnr among enterobacterial clinical isolates carrying ESBLs between 2003 and 2004 in Barcelona was 4.9%. qnrA1 was the most prevalent, whereas only one qnrS and no qnrB were detected.