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Enterococcus faecium  E35048, a bloodstream isolate from Italy, was the first strain where the oxazolidinone resistance gene  optrA  was detected outside China. The strain was also positive for the oxazolidinone resistance gene  cfr . WGS analysis revealed that the two genes were linked (23.1 kb apart), being co-carried by a 41,816-bp plasmid that was named pE35048-oc. This plasmid also carried the macrolide resistance gene  erm (B) and a backbone related to that of the well-known  Enterococcus faecalis  plasmid pRE25 (identity 96%, coverage 65%). The  optrA  gene context was original,  optrA  being part of a composite transposon, named Tn 6628 , which was integrated into the gene encoding for the ζ toxin protein ( orf19  of pRE25). The  cfr  gene was flanked by two IS Enfa5  insertion sequences and the element was inserted into an  lnu (E) gene. Both  optrA  and  cfr  contexts were excisable. pE35048-oc could not be transferred to enterococcal recipients by conjugation or transformation. A plasmid-cured derivative of  E. faecium  E35048 was obtained following growth at 42°C, and the complete loss of pE35048-oc was confirmed by WGS. pE35048-oc exhibited some similarity but also notable differences from pEF12-0805, a recently described enterococcal plasmid from human  E. faecium  also co-carrying  optrA  and  cfr ; conversely it was completely unrelated to other  optrA - and  cfr -carrying plasmids from  Staphylococcus sciuri . The  optrA - cfr  linkage is a matter of concern since it could herald the possibility of a co-spread of the two genes, both involved in resistance to last resort agents such as the oxazolidinones.

Characterization of a Multiresistance Plasmid Carrying the optrA and cfr Resistance Genes From an Enterococcus faecium Clinical Isolate