The World Health Organization recently recognized the Republic of Kosovo as one of the highest consumers per capita of antibiotics for human use among non- European Union Eastern European countries; however, data are limited regarding antimicrobial usage and antimicrobial resistance in the livestock sector for this recently formed country. The objective of this study was to conduct the first nationwide survey of antimicrobial resistance phenotypes in indicator bacteria collected from dairy farms in Kosovo. Composite fecal samples were collected from 52 farms located within all 7 administrative districts of Kosovo in the summer of 2014. Isolation and characterization of the indicator bacteria Escherichia coli (n = 165) and Enterococcus spp. (n = 153) from these samples was achieved by culturing on selective/differential media with and without select antibiotics, followed by MALDI-TOF (matrixassisted laser desorption/ionization time-of-flight) mass spectrometry-based identification and antimicrobial susceptibility testing using the disk diffusion method. When no selective pressure was applied in culture-based isolation, the majority of E. coli and Enterococcus spp. collected were resistant to ≤1 of 16 and ≤2 of 12 antibiotics tested, respectively. In contrast, E. coli and Enterococcus spp. isolated using sub-minimum inhibitory concentrations of cefoxitin, ciprofloxacin, or erythromycin were typically resistant to at least one and often multiple antibiotic types, which primarily consisted of certain β-lactams, quinolones, sulfonamides, phenicols, and tetracyclines for E. coli isolates and macrolides, tetracyclines, and rifamycins for enterococci isolates
