ABSTRACTThis study describes the genotypic characteristics of a collection of 100 multidrug-resistant (MDR)Escherichia colistrains recovered from cattle and the farm environment in Ireland in 2007. The most prevalent antimicrobial resistance identified was to streptomycin (100%), followed by tetracycline (99%), sulfonamides (98%), ampicillin (82%), and neomycin (62%). Resistance was mediated predominantly bystrA-strB(92%),tetA(67%),sul2(90%),blaTEM(79%), andaphA1(63%) gene markers, respectively. Twenty-seven isolates harbored a class 1 integrase (intI1), whileqacEΔ1andsul1markers were identified in 25 and 26 isolates, respectively. The variable regions of these integrons contained aminoglycoside, trimethoprim, and β-lactam resistance determinants (aadA12,aadB-aadA1,blaOXA-30-aadA1,dfrA1-aadA1,dfrA7). Class 2 integrons were identified less frequently (4%) and contained the gene cassette arraydfrA1-sat1-aadA1. Resistance to ampicillin, neomycin, streptomycin, sulfonamide, and tetracycline was associated with transferable high-molecular-weight plasmids, as demonstrated by conjugation assays. A panel of virulence markers was screened for by PCR, and genes identified includedvt1, K5 in 2 isolates,papCin 10 isolates, and PAI IV536in 37 isolates. MDR commensalE. coliisolates from Irish cattle displayed considerable diversity with respect to the genes identified. Our findings highlight the importance of the commensal microflora of food-producing animals as a reservoir of transferable MDR.