| Summary: | During the last decade’s increase of antimicrobial resistance (AMR) in animals, animal-human transmission has become a major threat. Therefore, the present study aimed to evaluate the genetic basis of AMR in Gram-negative bacteria recovered from sheep and goats with respiratory disease. Nasal and ocular swabs were collected from 69 diseased animals, and 76 Gram-negative bacterial isolates were identified from 59 animals. All isolates were checked phenotypically for resistance and genotypically for different resistance mechanisms, including β-lactam, quinolone, and aminoglycoside resistance. Our results demonstrated that 9.2% (95% CI 4.5–17.8%) of the isolates were multidrug-resistant, with high resistance rates to β-lactams and quinolones, and 11.8% (95% CI 6.4–21%) and 6.6% (95% CI 2.8–14.5%) of the isolates were phenotypically positive for AmpC and ESBL, respectively. Genotypically, <i>bla</i><sub>TEM</sub> was the most identified β-lactamase encoding gene in 29% (95% CI 20–40%) of the isolates, followed by <i>bla</i><sub>SHV</sub> (14.5%, 95% CI 8.3–24.1%) and <i>bla</i><sub>CTX-M</sub> (4%, 95% CI 1.4–11%). Furthermore, 7.9% (95% CI 3.7–16.2%) of the isolates harbored plasmid-mediated quinolone resistance gene <i>qnrS</i>. Our study revealed for the first time to our knowledge high β-lactam and quinolone resistance associated with the bacteria recovered from sheep and one goat with respiratory disease. Furthermore, different antimicrobial resistant determinants were identified for the first time from animals in Africa, such as <i>bla</i><sub>LEN-13/55</sub>, <i>bla</i><sub>TEM-176</sub> and <i>bla</i><sub>TEM-198/214</sub>. This study highlights the potential role of sheep and goats in disseminating AMR determinants and/or resistant bacteria to humans. The study regenerates interest for the development of a One Health approach to combat this formidable problem.
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