Simple SummaryDirect and indirect exposure of antibiotics to pet animals has been shown to play a role in the transmission of antimicrobial-resistant bacteria. Our study from the United Arab Emirates found that a public health relevant antibiotic-resistant strain ofEscherichia coli(E. coli) bacteria is present in fecal swabs sampled from many healthy pet cats and dogs. The strain, known as extended-spectrum β-lactamases resistant (ESBL-R)E. coli, is resistant to many commonly used antibiotics, making it difficult to treat if it causes an infection. We found that 23.65% of the animals tested had ESBL-RE. coli. The bacteria were most commonly found in pets that had access to water in ditches and puddles. The study also found that 91% of the bacteria samples were multidrug resistant, meaning that they were resistant to multiple types of antibiotics. The findings from this study call for veterinarians to develop surveillance programs to monitor ESBL-RE. coliin pets and to reduce the risk of transmission to humans and the environment.AbstractExtended-spectrum β-lactamases resistant (ESBL-R)Escherichia coli(E. coli) has been reported from healthy and sick pets. However, data from Middle Eastern countries, including the United Arab Emirates (UAE), are minimal. This study provides the first evidence of ESBL-RE. colicarriage among pets in the UAE. A total of 148 rectal swabs were collected from domestic cats (n= 122) and dogs (n= 26) attending five animal clinics in the UAE. Samples were cultured directly onto selective agar, and suspected colonies were confirmed as ESBL-producing using phenotypic and molecular methods. Confirmed isolates were screened for their phenotypic resistance to twelve antimicrobial agents using the Kirby Bauer method. The owners of the pets completed a questionnaire at the time of sampling, and the data were used to identify risk factors. ESBL-RE. coliwas detected in rectal swabs of 35 out of 148 animals (23.65%) (95% confidence interval [CI]: 17.06–31.32). Multivariable logistic regression analysis identified cats and dogs with access to water in ditches and puddles as 3.71 (p-value = 0.020) times more likely to be positive to ESBL-RE. colithan those without access to open water sources. Ciprofloxacin resistance was evident in 57.14% (44/77) of the ESBL-RE. coliisolates. The percentage of resistance to azithromycin and cefepime was 12.99% (10/77) and 48.05% (37/77), respectively. TheblaCTX-Mgene was detected in 82% of the PCR-screened isolates (n= 50). Multidrug resistance (MDR) phenotypes were evident in 91% (70/77) of the isolates. In conclusion, ESBL-RE. coliwas detected at a noticeable rate among healthy pet cats and dogs in the UAE, and the majority are MDR to clinically important antimicrobials such as fluoroquinolones and 3rd and 4th generation cephalosporins. Our results call for strengthening antimicrobial stewardship among companion animal veterinarians in the UAE to reduce the potential transmission of ESBL-RE. colibetween pets, humans, and urban environments.