| Summary: | The aim of the present study was to characterize <i>Enterobacterales</i> resistant to 3rd and 4th generation cephalosporins, carbapenems and/or fluoroquinolones, isolated from dogs and cats with urogenital infections. In total, 36 strains (<i>Escherichia coli</i> (<i>n</i> = 28), <i>Klebsiella pneumoniae</i> (<i>n</i> = 3), <i>Serratia marcescens</i>, <i>Raoultella ornithinolytica</i>, <i>Proteus mirabilis</i>, <i>Citrobacter portucalensis</i> and <i>Enterobacter cloacae</i> (each <i>n</i> = 1)) were included in the present study, 28 from Austria and 8 from Serbia. Isolates were characterized by a polyphasic approach including susceptibility pheno- and genotyping and microarray-based assays. <i>Escherichia</i> (<i>E.</i>) <i>coli</i> isolates were additionally characterized by two-locus (<i>fum</i>C and <i>fim</i>H) sequence phylotyping and multi-locus sequence typing (MLST) of selected isolates. MLST of carbapenem-resistant <i>Enterobacter cloacae</i> isolates was also performed. Among <i>E. coli</i>, the most dominant phylogenetic group was B1 (27.8%), followed by C, (16.6%), A and Clade II (5.5% each), B2 and F (2.77% each). The most predominant β-lactam resistance genes were <i>bla</i><sub>TEM</sub> (70%) and <i>bla</i><sub>CTX-M</sub> (38.8%), <i>bla</i><sub>CMY</sub> (25%). <i>bla</i><sub>NDM</sub> was detected in one carbapenem-resistant <i>Enterobacter cloacae</i> ST114. The most common ST among selected <i>E. coli</i> was 744 (10.7% isolates). The pandemic clones ST131 and ST648 carrying CTX-M-15 were also detected. Remaining STs belonged to 469, 1287, 1463 and 1642. <i>E. coli</i> clonotyping revealed 20 CH types. Based on the presence of certain virulence genes, three isolates were categorized as ExPEC/UPEC. The most prevalent virulence factors were <i>fim</i>H detected in 61%, <i>iuc</i>D and <i>iss</i> both in 55%, <i>iro</i>N in 27.8%, <i>pap</i>C in 13.8% and <i>sat</i> in 8.3% isolates.
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