f Extended-spectrum-beta-lactamase (ESBL)-producing, AmpC beta-lactamase-producing, and plasmid-mediated quinolone resistance (PMQR) gene-positive strains of Escherichia coli were investigated in wintering rooks (Corvus frugilegus) from eight European countries. Fecal samples (n ؍ 1,073) from rooks wintering in the Czech Republic, France, Germany, Italy, Poland, Serbia, Spain, and Switzerland were examined. Resistant isolates obtained from selective cultivation were screened for ESBL, AmpC, and PMQR genes by PCR and sequencing. Pulsed-field gel electrophoresis and multilocus sequence typing were performed to reveal their clonal relatedness. In total, from the 1,073 samples, 152 (14%) cefotaxime-resistant E. coli isolates and 355 (33%) E. coli isolates with reduced susceptibility to ciprofloxacin were found. Eighty-two (54%) of these cefotaxime-resistant E. coli isolates carried the following ESBL genes: bla CTX-M-1 (n ؍ 39 isolates), bla CTX-M-15 (n ؍ 25), bla CTX-M-24 (n ؍ 4), bla TEM-52 (n ؍ 4), bla CTX-M-14 (n ؍ 2), bla CTX-M-55 (n ؍ 2), bla SHV-12 (n ؍ 2), bla CTX-M-8 (n ؍ 1), bla CTX-M-25 (n ؍ 1), bla CTX-M-28 (n ؍ 1), and an unspecified gene (n ؍ 1). Forty-seven (31%) cefotaxime-resistant E. coli isolates carried the bla CMY-2 AmpC beta-lactamase gene. Sixty-two (17%) of the E. coli isolates with reduced susceptibility to ciprofloxacin were positive for the PMQR genes qnrS1 (n ؍ 54), qnrB19 (n ؍ 4), qnrS1 and qnrB19 (n ؍ 2), qnrS2 (n ؍ 1), and aac(6=)-Ib-cr (n ؍ 1). Eleven isolates from the Czech Republic (n ؍ 8) and Serbia (n ؍ 3) were identified to be CTX-M-15-producing E. coli clone B2-O25b-ST131 isolates. Ninety-one different sequence types (STs) among 191 ESBL-producing, AmpC-producing, and PMQR gene-positive E. coli isolates were determined, with ST58 (n ؍ 15), ST10 (n ؍ 14), and ST131 (n ؍ 12) predominating. The widespread occurrence of highly diverse ESBL-and AmpC-producing and PMQR gene-positive E. coli isolates, including the clinically important multiresistant ST69, ST95, ST117, ST131, and ST405 clones, was demonstrated in rooks wintering in various European countries.T he incidence of bacteria resistant to cephalosporins and fluoroquinolones is growing steadily and constitutes a serious risk for human and animal health. The major mechanism conferring resistance to cephalosporins is mediated by extended-spectrum beta-lactamases (ESBLs) and AmpC beta-lactamases (1). Although plasmid-mediated quinolone resistance (PMQR) genes confer only a low level of resistance to fluoroquinolones and resistance is mainly caused by point mutations of the quinolone resistance-determining region (QRDR), coding for gyrase and topoisomerase (2), the interaction between mutations in the QRDR and PMQR genes leads to higher levels of resistance to fluoroquinolones (3).Wild animals that do not come directly into contact with antibiotics are affected by their use in human and veterinary medicine. The close proximity of wild animals with humans and domestic animals plays an impo...