ᰔThe bla NDM-1 carbapenemase gene has now been identified among many different enterobacterial species from many countries (1,7,(10)(11)(12)(13)(14). Its current spread has often been associated with importations from the Indian subcontinent.Here we report the case of a 70-year-old man admitted to the intensive care unit of a hospital in Bonn, Germany, in December 2009. This patient had been hospitalized in India 3 months before as a consequence of a trauma that made him paraplegic. An acute appendicitis associated with paralytic ileus was diagnosed at the Bonn hospital. The surgical intervention was successful, but the patient remained on a ventilator and screening for colonization produced growth of a multidrug-resistant Escherichia coli isolate from his tracheal secretions. Since no symptoms of infection occurred, no antibiotic treatment was initiated, and the patient was shortly discharged.Antimicrobial drug susceptibility testing of the E. coli RKI isolate was performed by broth microdilution according to the guidelines of the CLSI (3). E. coli RKI was resistant to all -lactams, including carbapenems, with MICs of imipenem and meropenem being Ͼ32 g/ml; to all aminoglycosides; and to nitrofurantoin and sulfonamides but remained susceptible to tigecycline and fosfomycin, and the MIC of colistin was 0.5 g/ml. PCR and sequencing were performed in searching of different carbapenemase and extended-spectrum -lactamase (ESBL) genes and also for bla OXA genes as described previously (5, 9). These procedures showed that E. coli RKI harbored the bla NDM-1 , bla CTX-M-15 , bla TEM-1 , bla OXA-1 , and bla OXA-2 genes.In addition, PCR performed as described previously (4, 6, 11) revealed that E. coli RKI harbored the aac(6Ј)1b-cr gene, encoding resistance to aminoglycosides and reduced susceptibility to ciprofloxacin, together with the 16S rRNA methylase gene rmtC, conferring high-level resistance to all aminoglycosides, although no qnr gene was detected.Transfer of the bla NDM-1 gene was attempted by conjugation or electrotransformation (11) into a sodium azide-resistant E. coli J53 recipient strain, but the effort was unsuccessful. S1 nuclease pulsed-field gel electrophoresis (PFGE) analysis, performed as previously described (1), revealed two plasmids in E. coli RKI (120 and 70 kb, respectively) that did not hybridize with a bla NDM-1 -specific probe. Investigation of the bla NDM-1 genetic environment by PCR mapping using primers designed according to previously identified bla NDM-1 -associated structures (11) revealed that the bla NDM-1 genetic context in E. coli RKI was different from those previously identified, further underlining that the current dissemination of bla NDM-1 was not associated with a single specific genetic structure.Multilocus sequence typing (MLST) (http://mlst.ucc.ie /mlst/dbs/Ecoli) and PCR-based phylogroup analysis (2) identified E. coli RKI as an ST101 strain belonging to phylogroup B1. Interestingly, the NDM-1-positive E. coli 271 strain recently identified from Australia (11) with a link ...
