In this study, we analyzed the antimicrobial resistance properties and T antigenic types of 511 isolates collected in Lisbon district, Portugal, from throat swabs of healthy subjects (n=341), during 2000-2002 and from diverse infection sites (n=170) of outpatients and inpatients, during 1999-2002. Erythromycin resistance was higher in tonsillitis/pharyngitis (27.4%) and skin infection isolates (21.1%), than in carriage and invasive isolates (

During 2000-2007 in Lisbon, we identified 45 bacitracin-resistant Streptococcus pyogenes isolates among 1629 isolates: 24 from oropharyngeal healthy carriers (out of 1026), 21 from patients with noninvasive infections (out of 559) and zero from invasive infections (out of 44). Forty-four of those isolates, mainly of colonization, are low-level bacitracin-resistant members of the cMLS(B)-macrolide-resistant and tetracycline-susceptible emm28/ST52 clone previously detected in Europe, but only among clinical samples. One high-level bacitracin-resistant isolate, associated with a tonsillitis/pharyngitis episode, is cMLS(B)-macrolide-resistant and tetracycline-resistant member of the emm74/ST120 lineage, which was not previously known to include bacitracin-resistant isolates. The bcrABDR operon encoding an ATP-binding cassette transporter in Enterococcus faecalis was not detected among these bacitracin-resistant S. pyogenes strains. Virulence profiling indicated that genes coding for exotoxins and superantigens seem to be clone specific. This study provides an increased knowledge about specific bacitracin-resistant S. pyogenes strains, which may be useful in future investigations aiming to understand the mechanism(s) leading to bacitracin resistance and the cause(s) for differences in colonization and/or dissemination potential.

We describe 66 ciprofloxacin-nonsusceptible Streptococcus pyogenes isolates recovered from colonized and infected children. The ParC S79A substitution was frequent and associated with the emm6/sequence type 382 (emm6/ST382) lineage. The ParC D83G substitution was detected in two isolates (emm5/ST99 and emm28/ST52 lineages). One isolate (emm89/ST101) had no quinolone resistance-determining region codon substitutions or other resistance mechanisms. Five of 66 isolates were levofloxacin resistant. Although fluoroquinolones are not used in children, they may be putative disseminators of fluoroquinolone-nonsusceptible strains in the community.Streptococcus pyogenes clinical isolates with reduced susceptibility to fluoroquinolones (1,3,8,14,16,18,27,29) or with high-level resistance (15,16,(23)(24)(25)28) have been described previously, and the reduced susceptibility to fluoroquinolones is mediated by point mutations in the quinolone resistancedetermining region (QRDR) of the parC gene (3, 16, 18) whereas high-level resistance has been associated with mutations in the QRDRs of both parC and gyrA genes (23, 28). To the best of our knowledge, there are no reports documenting the prevalence and characterization of fluoroquinolone-nonsusceptible S. pyogenes associated with asymptomatic colonization. Since 1999 and 2000, we have been collecting S. pyogenes isolates from pediatric patients from different clinical origins and from carriers for the surveillance of antimicrobial susceptibility and for the epidemiological characterization of the isolates. This study aimed to describe the prevalence of ciprofloxacin-nonsusceptible S. pyogenes isolates from colonized and infected Portuguese children from 1999 to 2006 and to characterize the associated clones and resistance mechanisms.Strains and antibiotic susceptibility. A total of 1,354 nonduplicated S. pyogenes isolates were collected from children in the Lisbon area in Portugal; 901 were associated with asymp-

The asymptomatic oropharyngeal colonization rate by Streptococcus pyogenes was 10.7% in children (901 among 8,405 children 0-16 years old) and 3.3% in adults (37 among 1,126 households of children) in the Lisbon area during 2000-2006. Macrolide-resistant S. pyogenes from children (n = 149) was variable with time: 9.8-10.7% in 2000-2002, 28.1% in 2003, 19.6-2.7% in 2004-2005 and 14.6% in 2006. Eight lineages (97.3% of isolates) were identified based on at least 80% similarity of PFGE patterns, T types, emm types and multilocus sequence types (ST). The elevated frequency of macrolide resistance was associated with M phenotype lineages I (emm12/ST36) and V (emm4, emm75/ST39 and a novel emmstMrp6 type) and with one cMLS(B) lineage IV (emm28/ST52) known to be associated with upper respiratory tract and invasive infections. Significant associations (p < 0.05) between emm type/virulence genotype were found, such as emm1/speA (+) ssa (-), emm4/ssa (+) prtF1 (+), emm12/speA (-) ssa (-). The high prevalence (>20%) of speC, prtF1 or ssa was probably caused either by clonal dissemination (speC), or to horizontal gene transfer events (prtF1 and ssa). This report contributes to a better understanding of the molecular epidemiology and evolution of macrolide-resistant S. pyogenes causing symptom-free oropharyngeal colonization. These colonizing strains carry macrolide resistance and virulence genes capable of being transferred to other bacterial species sharing the same niche.

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