Emergence and spread of a multidrug-resistant Acinetobacter baumannii clone producing both the carbapenemase OXA-23 and the 16S rRNA methylase ArmA.

Acinetobacter baumannii is a Gram-negative organism reported worldwide as a cause of health-care-associated infections, particularly in intensive care units (ICUs). The aim of this study is to describe the emergence and spread of carbapenem-resistant A. baumannii (CRAB) isolates in hospitalized patients. From March to November 2009, multidrug-resistant CRAB isolates were obtained from 21 patients hospitalized in different wards (mostly ICUs). Antimicrobial susceptibility was determined by using the Etest method. Carbapenem and aminoglycoside resistance determinants were studied by PCR and sequencing. Genetic relatedness was investigated by pulsed-field gel electrophoresis and multiplex PCR identification of sequence groups. Clinical records of patients were examined retrospectively. CRAB isolates were consistently resistant to multiple drugs including fluoroquinolones and aminoglycosides, whereas they retained a susceptibility to colistin. Molecular analysis revealed that 19 of the 21 CRAB isolates belonged to a single clone producing both the carbapenemase OXA-23 and the 16S rRNA methylase ArmA. Based on clinical data, the patients included in the study were classified as infected (n=13) or colonized (n=8). Colistin alone or in combination with ampicillin-sulbactam was administered to 11 of the 13 infected patients. A complete or partial response was obtained in eight cases, whereas a failure to respond was observed in one patient and a relapse was observed in two patients. An A. baumannii clone producing both OXA-23 and ArmA has been identified as an emerging and rapidly spreading pathogen. To our knowledge, this is the first report of the ArmA enzyme in A. baumannii in Italy and is the first report of hospital dissemination of A. baumannii carrying both bla(OXA-23) and armA genes.

Prevalence of plasmid-mediated quinolone resistance genes in uropathogenic Escherichia coli isolated in a teaching hospital of northern Italy.

A retrospective study was conducted to determine the prevalence of plasmid-mediated quinolone resistance (PMQR) determinants in uropathogenic Escherichia coli isolated from inpatients and outpatients in a teaching hospital of northern Italy. The presence of qnrA, qnrB, qnrS, aac(6')-Ib-cr, and qepA was evaluated in 76 and 72 nalidixic acid-resistant E. coli, isolated in 2004 and 2006, respectively. Positivity for the aac(6')-Ib-cr gene was demonstrated in 3 of 76 (3.9%) and 8 of 72 (11%) isolates, respectively; no other PMQR determinant was found. All aac(6')-Ib-cr-positive strains also showed two point mutations in the gyrA and parC genes. Most aac(6')-Ib-cr-positive isolates demonstrated the contemporary presence of bla(CTX-M-15), bla(OXA-1/30), and bla(TEM-1) genes and 4/11 harbored a class 1 integron with a dfrA17-aadA5 gene cassette arrangement. Interestingly, all aac(6')-Ib-cr-positive isolates belonged to B2 phylogenetic group, O25b antigen type, multi locus sequence type 131, and to a cluster of approximately 70% similarity level by pulsed-field gel electrophoresis (PFGE). These findings suggest the circulation of the previously described intercontinentally spreading E. coli O25:H4-ST131 clone in our geographical area since 2004. Hybridization studies of the PFGE profiles showed the aac(6')-Ib-cr gene to be associated with different molecular weight bands (40-350 kb) and interestingly aac(6')-Ib-cr chromosomal integration was demonstrated in one strain by I-Ceu I method. This represents the first report to investigate the presence and diffusion of PMQR determinants in northern Italy and to describe aac(6')-Ib-cr chromosomal integration in E. coli.