 This study aims to fully characterize the genomes of multi-driven resistant, NDR, and antimicrobial susceptible Escherichia coli strains isolated from pre-winged calves, post-winged calves, dry cows, and lactating cows on commercial dairy operations. The results indicate that both susceptible and resistant E. coli isolates recovered from animals on commercial dairy operations are highly diverse and encode a large pool of virulence factors. In total, 838 transferable antimicrobial resistance genes, ARGs, were detected, with genes conferring resistance to aminoblycosides being the most common. Multiple sequence types associated with mild to severe human gastrointestinal and extraintestinal infections were identified. A Fisher's exact test identified 619 genes that were significantly enriched in NDR isolates and 147 genes that were significantly enriched in susceptible isolates. Significantly enriched genes in NDR isolates include the iron scavenging aerobactin synthesis and receptor genes, UKBCDIDA, the CIT-ABCD system, the PFIMBRE E-PAP genes, muonositol catabolism, IOL-Aptigiatia, and ascorbate transport genes. The results demonstrate a highly diverse population of E. coli and commercial dairy operations, some of which encode virulence genes responsible for severe human infections and resistance to antibiotics of human health significance. Further, the enriched accessory genes in NDR isolates, aerobactin, CIT, PFIMBRE, and muonositol catabolism in ascorbate transport genes represent potential targets for reducing colonization of antimicrobial resistant bacteria in the calf gut. This article was authored by Brad J. Haley, C. N. Woo Kim, Sarah J. Salaheen, and others. We're article.tv, links in the description below.