| Summary: | The occurrence of antimicrobial resistance in commensal strains of <i>Escherichia coli</i> and <i>Staphylococcus</i> spp. was investigated in 320 samples collected from patients and the environment of a veterinary university hospital—specifically, the consultation area (CA) and intensive care unit (ICU). <i>E. coli</i> was isolated in 70/160 samples (44%), while <i>Staphylococcus</i> spp. were isolated in 110/160 (69%) samples. The occurrence of multidrug-resistant (MDR) isolates from CA and ICU admission were similar for <i>E. coli</i> (1/12 (8%) versus 4/27 (15%), respectively) and <i>Staphylococcus</i> spp. (10/19 (53%) versus 26/50 (52%), respectively). MDR <i>E. coli</i> isolates increased significantly at hospital discharge (18/31; 58%; <i>p</i> = 0.008). Antimicrobial treatment administered during hospitalization was a risk factor for carriage of MDR <i>E. coli</i> (OR, 23.9; 95% CI: 1.18–484.19; <i>p</i> = 0.04) and MDR <i>Staphylococcus</i> spp. (OR, 19.5; 95% CI 1.30–292.76; <i>p</i> = 0.02), respectively. The odds ratio for MDR <i>E. coli</i> was 41.4 (95% CI 2.13–806.03; <i>p</i> = 0.01), if the administration of fluoroquinolones was evaluated. The <i>mecA</i> gene was detected in 19/24 (79%) coagulase-positive <i>Staphylococcus</i> spp. isolates resistant to oxacillin. High rates of MDR <i>Staphylococcus</i> spp. were reported. Hospitalization in the ICU and antimicrobial treatment were risk factors for colonization by MDR commensal bacteria.
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