AimsThe study aims to demonstrate the antimicrobial and disinfectant resistance phenotypes and genotypes of Escherichia coli isolates obtained from giant pandas (Ailuropoda melanoleuca). Methods and ResultsAntimicrobial testing was performed according to the standard disk diffusion method. The minimal inhibitory concentrations (MICs) of disinfectants were determined using the agar dilution method. All isolates were screened for the presence of antimicrobial and disinfectant resistance genes and further analysed for genetic relatedness by pulse-field gel electrophoresis (PFGE). Results showed that 466% of the isolates were resistant to at least one antimicrobial. Escherichia coli isolates showed resistance to fewer antimicrobials as panda age increased. Among antimicrobial-resistant E.coli isolates, the antimicrobial resistance genes blaCTX-M (882%) and sul1 (923%) were most prevalent. The disinfectant resistance genes emrE, ydgE/ydgF, mdfA and sugE(c) were commonly present (682-989%), whereas qac and sugE(p) were relatively less prevalent (0-213%). The frequencies of resistance genes tended to be higher in E.coli isolated in December than in July, and PFGE profiles were also more diverse in isolates in December. The qacE1 and sugE(p) genes were higher in adolescent pandas than in any other age groups. PFGE revealed that antimicrobial resistance correlated well with sampling time and habitat. ConclusionsThis study demonstrated that antimicrobial and disinfectant resistance was common in giant panda-derived E.coli, and the antimicrobial resistance was associated with sampling time and habitat. Escherichia coli could serve as a critical vector in spreading disinfectant and antimicrobial resistance. Significance and Impact of the StudyThis is the first study that demonstrated the phenotypic and genetic characterizations of antimicrobial and disinfectant resistance in E.coli isolates from more than 60 giant pandas. Frequent transfer of pandas to other cages may lead to the dissemination of antimicrobial resistance. The study highlights the need for regularly monitoring the antimicrobial and disinfectant resistance in bacteria from giant pandas.