This considerably exceeded the rate reported by previous studies

This considerably exceeded the rate reported by previous studies (Gibb et al., 1992; Appelmelk et al., 1994; Amor et al., 2000; Gibbs et al., 2004). As the core type distribution among non-ESBL-producing strains (3.7%) was similar to those found earlier (Table 1), and as the production of ESBL is, at least partly, a clonal phenomenon (Woodford et al., 2011), the possible clustering

of the 58 K-12 core PCR-positive isolates was investigated. We found that 54 of these strains (93.1%) carried the rfbO25b gene with the O25 serogroup also confirmed by slide agglutination. All strains belonged to the B2 phylogenetic group. All isolates, except two, were ESBL-producing strains. Fifty-two CHIR-99021 purchase of the 54 K-12 learn more core and rfbO25b-positive strains were typable by PFGE exhibiting 18 pulsotypes (10 clusters with 2–11 members and 8 singletons) (Fig. 1). Twenty-four selected isolates representing all pulsotypes were submitted to MLST and found to belong to the rapidly spreading, often multidrug resistant ST131 clone (Fig. 1). To rule out that the presence of the K-12 core-specific genes was restricted to Emirati UTI isolates of the O25 ST131 group, ten independent representatives of this clone isolated in Hungary from UTI (five strains) and BSI (five strains) in 2008 and 2009, respectively, were also tested. Importantly, all these strains were also positive with the

K-12 core-specific PCR (Fig. 1). Next, we determined the DNA sequence of the entire waa locus (Heinrichs et al., 1998) of one of the O25-ST131 isolates from our collection (#81009). The resulting > 16-kb sequence (GenBank JQ241150) covered the 15 K-12 core genes (Muller-Loennies et al., 2007) between 4-Aminobutyrate aminotransferase the kbl and the coaD genes flanking the waa locus. As expected, based on the PCR results, individual gene sequences displayed extensive homology to their respective homologues in the prototype K-12 commensal strain, MG1655 (Table 2). Comparison of the deduced amino acid sequences of the various Waa proteins of the ST131 O25 strain #81009 revealed ≥ 90% identities

with their counterparts in MG1655 with the exception of WaaQ, exhibiting a 71% homology, only (Table 2). This enzyme of strain #81009, however, was 99% identical to its counterparts found in strains with core types R1, R3, and R4 (Table 2), while the WaaQ protein encoded by the MG1655 allele was identical to that of a representative R2 strain, F632. Because the function of this protein as a heptosyltransferase is completely conserved in all core types (Muller-Loennies et al., 2007), we surmise that this sequence variation is unlikely to have functional consequences. An almost 100% identity (except two nucleotide differences resulting in a single amino acid mismatch in WaaB) of the entire waa locus of the strain #81009 was found with that of a commensal fecal isolate SE15 of the B2 phylogenetic group (Toh et al., 2010) (Table 2).

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