Description of an original integron encompassing blaVIM-2, qnrVC1 and genes encoding bacterial group II intron proteins in Pseudomonas aeruginosa.

OBJECTIVES: A burn unit of a hospital in Tunis underwent an endemic situation caused by imipenem-resistant Pseudomonas aeruginosa. For nine non-repetitive isolates of a clonal VIM-2-producing strain, the blaVIM-2 genetic background was characterized and the associated qnrVC1 gene molecularly analysed. METHODS: The imipenem resistance mechanism was investigated by phenotypic and molecular tests, and resistance transfer was studied by conjugation and transformation experiments. The integron's structure was characterized by sequencing, and qnrVC1 expression was explored after cloning experiments. RESULTS: The nine VIM-2-producing strains were collected from eight patients and one environmental sample. All transfer assays failed, suggesting a chromosomal location of blaVIM-2. This latter was found to be part of a class 1 integron of ∼7500 bp, which also contains blaOXA-2, aadA1 and two copies of the aadB, arr-6 and qnrVC1 genes. qnrVC1 exhibited higher homology with the chromosomally encoded qnr genes of Vibrionaceae than with plasmid-mediated qnr genes of Enterobacteriaceae. The qnrVC1 gene cassette possesses a promoter allowing its expression, and it conferred decreased fluoroquinolone susceptibility to Escherichia coli. Additionally, on the same integron, genes encoding an uncommon group IIC-attC intron were detected. CONCLUSIONS: A VIM-2-producing P. aeruginosa outbreak led us to characterize an integron harbouring a qnrVC1 cassette and a new group IIC-attC intron. This is the first known description of a qnr determinant in a P. aeruginosa strain. Its presence conferred a low level of resistance to quinolones in E. coli, which might favour the emergence of highly resistant mutants.

Evolution of an incompatibility group IncA/C plasmid harboring blaCMY-16 and qnrA6 genes and its transfer through three clones of Providencia stuartii during a two-year outbreak in a Tunisian burn unit.

During a 2-year period in 2005 and 2006, 64 multidrug-resistant Providencia stuartii isolates, including 58 strains from 58 patients and 6 strains obtained from the same tracheal aspirator, were collected in a burn unit of a Tunisian hospital. They divided into four antibiotypes (ATB1 to ATB4) and three SmaI pulsotypes (PsA to PsC), including 49 strains belonging to clone PsA (48 of ATB1 and 1 of ATB4), 11 strains to clone PsB (7 of ATB2 and 4 of ATB3), and 4 strains to clone PsC (ATB3). All strains, except for the PsA/ATB4 isolate, were highly resistant to broad-spectrum cephalosporins due to the production of the plasmid-mediated CMY-16 ß-lactamase. In addition, the 15 strains of ATB2 and ATB3 exhibited decreased quinolone susceptibility associated with QnrA6. Most strains (ATB1 and ATB3) were gentamicin resistant, related to an AAC(6')-Ib' enzyme. All these genes were located on a conjugative plasmid belonging to the incompatibility group IncA/C(2) of 195, 175, or 100 kb. Despite differences in size and in number of resistance determinants, they derived from the same plasmid, as demonstrated by similar profiles in plasmid restriction analysis and strictly homologous sequences of repAIncA/C(2), unusual antibiotic resistance genes (e.g., aphA-6), and their genetic environments. Further investigation suggested that deletions, acquisition of the ISCR1 insertion sequence, and integron cassette mobility accounted for these variations. Thus, this outbreak was due to both the spread of three clonal strains and the dissemination of a single IncA/C(2) plasmid which underwent a remarkable evolution during the epidemic period.