Genetic Characterization of a Linezolid- and Penicillin-Resistant Enterococcus hirae Isolate Co-Harboring poxtA and pbp5fm

Linezolid and penicillin are critical for treating multidrug resistant (MDR) Gram-positive infections, but the emergence of resistance to both seriously threatens public health. Here, we first report the cocarrying poxtA (oxazolidinone resistance) and pbp5fm (β-lactam resistance) genes by the plasmid in a strain of Enterococcus hirae HDC14-2 derived from porcine. The isolate also exhibits MDR phenotypes to phenicols, oxazolidinones, tetracyclines, β-lactams, aminoglycosides, macrolides, and lincosamides. Whole-genome sequencing (WGS) revealed these resistance genes, along with tet(L), tet(M), catA, erm(B), aac(6)-aph(2”), aadE, spw, lsa(E), lnu(B), sat4, and aphA3, were clustered in a novel MDR region flanked by IS1216 elements on plasmid pHDC14-2.133K. This IS1216-bounded MDR region formed translocatable units (TUs), including an IS1216-poxtA TU that was also identified on a secondary plasmid, pHDC14-2.27K. Functional assays demonstrated the excisability and mobility of these TUs, indicating its potential ability integration into other plasmids or chromosomes. Critically, electrotransformation confirmed the transfer of pHDC14-2.27K (poxtA-carrying) to Enterococcus faecalis JH2-2, with retained TU activity and minimal fitness cost. This study provides the evidence of colocalized poxtA and pbp5fm on plasmids in enterococci, highlighting their role in disseminating pan-resistance among bacteria. Although E. hirae is not an important pathogenic bacterium to humans and animals, but its potential risk to horizontally spread of these resistance genes important in medicine still cannot be ignored.