Co-existence of the oxazolidinone resistance genes cfr and optrA on two transferable multi-resistance plasmids in one Enterococcus faecalis isolate from swine.

ABSTRACT

OBJECTIVES: To identify and characterize oxazolidinone resistance genes cfr and optrA in enterococcal isolates. METHODS: Two hundred and ninety-three enterococcal isolates were screened for the presence of cfr and optrA by polymerase chain reaction. The transferability of cfr and optrA was examined by conjugation. S1 nuclease pulsed-field gel electrophoresis and Southern blotting were used to identify the location of cfr and optrA. One Enterococcus faecalis isolate carrying both cfr and optrA was sequenced in full. RESULTS: cfr and optrA were detected in 16 (5.5%) and 170 (58.0%) enterococcal isolates, respectively. Sixteen enterococcal isolates (E. faecalis n=13, Enterococcus avium n=2, Enterococcus mundtii n=1) carried both cfr and optrA. The cfr-carrying fragment between res and theta in plasmid p4 showed 98.9% identity to the corresponding region of plasmid pEF120805 from vancomycin-resistant Enterococcus faecium. The optrA-carrying segment between tnpB and optrA in plasmid p1 showed >99.9% identity to the corresponding region of genomic DNA from E. faecalis A101. Plasmid p4 and plasmid p1 were simultaneously conjugated to E. faecalis JH2-2. CONCLUSIONS: One hundred and seventy optrA-positive enterococci were identified in 293 enterococcal isolates from swine and the farm environment. The co-existence of cfr and optrA in E. avium and E. mundtii has been identified previously. cfr and optrA were identified on two new conjugative plasmids from one E. faecalis isolate. The optrA-carrying segment (IS1216E-optrA-IS1216E) was reported initially. Among different types of enterococcal plasmids, ISEnfa5 and IS1216E elements may play a vital role in the dissemination of cfr and optrA, respectively.