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For a One-Health investigation of antimicrobial resistance (AMR) in  Enterococcus  spp., isolates from humans and beef cattle along with abattoirs, manured fields, natural streams, and wastewater from both urban and cattle feedlot sources were collected over two years. Species identification of  Enterococcus  revealed distinct associations across the continuum. Of the 8430 isolates collected,  Enterococcus faecium  and  Enterococcus faecalis  were the main species in urban wastewater (90%) and clinical human isolates (99%);  Enterococcus hirae  predominated in cattle (92%) and feedlot catch-basins (60%), whereas natural streams harbored environmental  Enterococcus  spp. Whole-genome sequencing of  E. faecalis  (n = 366 isolates) and  E. faecium  (n = 342 isolates), revealed source clustering of isolates, indicative of distinct adaptation to their respective environments. Phenotypic resistance to tetracyclines and macrolides encoded by  tet(M)  and  erm(B)  respectively, was prevalent among  Enterococcus  spp. regardless of source. For  E. faecium  from cattle, resistance to β-lactams and quinolones was observed among 3% and 8% of isolates respectively, compared to 76% and 70% of human clinical isolates. Clinical vancomycin-resistant  E. faecium  exhibited high rates of multi-drug resistance, with resistance to all β-lactam, macrolides, and quinolones tested. Differences in the AMR profiles among isolates reflected antimicrobial use practices in each sector of the One-Health continuum.

Surveillance of Enterococcus spp. reveals distinct species and antimicrobial resistance diversity across a One-Health continuum