Virulome Landscape of Multidrug-Resistant <i>Escherichia coli</i> Across Human, Animal, and Environmental Reservoirs

<b>Background/Objectives:</b><i>Escherichia coli</i> (<i>E. coli</i>) spans commensal, intestinal pathogenic, and extraintestinal pathogenic lineages distributed across human, animal, and environmental reservoirs, yet the extent to which virulence architectures are shared across these compartments remains incompletely understood. Using a One Health framework, we profiled putative virulence determinants in pooled multidrug-resistant (MDR) <i>E. coli</i> source groups representing human, animal, and environmental sectors. <b>Methods:</b> Virulence genes were predicted with VirulenceFinder, and presence-absence profiles were integrated to define functional composition, sector overlap, source-group distribution breadth, and pathotype-associated signatures. Predicted pathogenic potential was assessed with PathogenFinder and compared with pathogenic family richness. <b>Results:</b> Overall, 114 putative virulence genes were detected, with adhesion/colonization functions dominating the virulome (33/114), followed by toxin-associated genes (12/114). A conserved core of 50 virulence genes was shared across all three sectors, including determinants linked to serum resistance (<i>iss</i>, <i>ompT</i>, <i>traT</i>), adhesion (<i>csgA</i>, <i>fimH</i>), stress adaptation (<i>terC</i>), and iron acquisition (<i>sitA</i>, <i>iutA</i>, <i>fyuA</i>). ExPEC-associated determinants were most numerous in environmental source groups (n = 52), whereas diarrheagenic <i>E. coli</i> markers were most frequent in animal-associated groups (n = 42). LEE-associated effectors were infrequent and largely absent from human source groups. Despite ecological differences in virulence composition, pathogenicity scores remained consistently high across sectors (0.83-0.92) and showed no significant association with pathogenic family richness (Spearman's ρ = 0.197, <i>p</i> = 0.392). <b>Conclusions:</b> Within the limits of pooled source-group analysis, these findings suggest that MDR <i>E. coli</i> across One Health compartments shares a broadly distributed, ExPEC-associated virulence repertoire overlaid with sector-specific pathotype signals, underscoring the value of integrated genomic surveillance while highlighting the need for isolate-resolved analysis.