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Salmonella enterica  and  Escherichia coli  can inhabit humans and animals from multiple origins. These bacteria are often associated with gastroenteritis in animals, being a frequent cause of resistant zoonotic infections. In fact, bacteria from animals can be transmitted to humans through the food chain and direct contact. In this study, we aimed to assess the antibiotic susceptibility of a collection of  S. enterica  and  E. coli  recovered from animals of different sources, performing a genomic comparison of the plasmid-mediated quinolone resistance (PMQR)-producing isolates detected. Antibiotic susceptibility testing revealed a high number of non-wild-type isolates for fluoroquinolones among  S. enterica  recovered from poultry isolates. In turn, the frequency of non-wild-type  E. coli  to nalidixic acid and ciprofloxacin was higher in food-producing animals than in companion or zoo animals. Globally, we detected two  qnrS1  and two  aac(6 ′ )-Ib-cr  in  E. coli  isolates recovered from animals of different origins. The genomic characterization of QnrS1-producing  E. coli  showed high genomic similarity (O86:H12 and ST2297), although they have been recovered from a healthy turtle dove from a Zoo Park, and from a dog showing symptoms of infection. The  qnrS1  gene was encoded in a IncN plasmid, also carrying  bla  TEM−1− containing Tn 3 . Isolates harboring  aac(6 ′ )-Ib-cr  were detected in two captive bottlenose dolphins, within a time span of two years. The additional antibiotic resistance genes of the two  aac(6 ′ )-Ib-cr- positive isolates ( bla  OXA−1 ,  bla  TEM−1 , bla  CTX−M−15 ,  catB3, aac(3)-IIa , and  tetA ) were enclosed in IncFIA plasmids that differed in a single transposase and 60 single nucleotide variants. The isolates could be assigned to the same genetic sublineage—ST131  fimH 30-Rx (O25:H4), confirming clonal spread. PMQR-producing isolates were associated with symptomatic and asymptomatic hosts, which highlight the aptitude of  E. coli  to act as silent vehicles, allowing the accumulation of antibiotic resistance genes, mobile genetic elements and other relevant pathogenicity determinants. Continuous monitoring of health and sick animals toward the presence of PMQR should be strongly encouraged in order to restrain the clonal spread of these antibiotic resistant strains.

QnrS1- and Aac(6′)-Ib-cr-Producing Escherichia coli among Isolates from Animals of Different Sources: Susceptibility and Genomic Characterization