Few studies have addressed drug resistance of Enterobacterales in rural communities in developing countries. This study aimed to determine the coexistence of extended-spectrum β-lactamase (ESBL) and carbapenemase genes inEscherichia coliandKlebsiella pneumoniaestrains carrying themcr-1gene in rural communities in Ecuador from healthy humans and their backyard animals. Sixty-two strains, thirtyE. coliand thirty-twoK. pneumoniaestrains carrying themcr-1gene were selected from a previous study. PCR were performed for the presence of ESBLs and carbapenemase genes. The strains were further characterized, and the genetic relationship was studied with multi-locus sequencing typing (MLST) of seven housekeeping genes. Fifty-nine of the sixty-twomcr-1isolates (95%) harbored at least on β-lactam resistance gene. The most prevalent ESBL genes were theblaTEMgenes (present in in 80% of theE. colistrains) and theblaSHVgene (present in 84% of theK. pneumoniaestrains). MSLT analysis revealed 28 different sequence types (ST); 15 forE. coliand 12 forK. pneumoniae, with most ST never described in humans and animals. The coexistence ofmcr-1and β-lactams resistant genes inE. coliandK. pneumoniaestrains is alarming and threatens the efficacy of last-resort antibiotics. Our findings highlight backyard animals as a reservoir ofmcr-1/β-lactams resistant genes.