In a very remote human community of the Peruvian Amazonas with minimal antibiotic exposure, high levels of acquired resistance to the oldest antibiotics (ampicillin, tetracycline, trimethoprim/sulfamethoxazole, streptomycin and chloramphenicol) were detected in commensal Escherichia coli, with remarkable diversity of resistant clones and of resistance genes and plasmids. This pattern was similar overall to that previously observed in a very remote community of Bolivia. It was also similar to that observed in the nearest urban area, except for a lower dominance of resistant isolates and the absolute lack of quinolone resistance in the remote community. Present findings suggest that antibiotic resistance observed in remote communities with minimal antibiotic exposure is a general phenomenon contributed by complex mechanisms and provide new insights into the mechanisms involved in this phenomenon.