Antimicrobial-resistant bacteria were isolated from muscoid dipterans collected at five different areas of Rio de Janeiro city, in proximity to hospitals. Extracts obtained by maceration of flies were diluted and used as inocula for different culture media, with or without antibiotic (ceftriaxone 1 mg/L) supplementation. Purified isolates were submitted to antimicrobial susceptibility testing (AST). Bacterial identification was performed by MALDI TOF Microflex LT (Bruker Daltonics). A total of 197 bacterial strains were obtained from 117 dipterous muscoids. Forty-two flies (35.9%) carried bacteria resistant to at least one antimicrobial, while 7 insects (5.9%) carried multidrug-resistant bacteria (MDR), which were all members of the family Enterobacteriaceae. Among 10 MDR bacteria (5%), 5 strains (2,5%) were positive by PCR for one or more of the following antibiotic resistance genes: aac(6’)-Ib, blaTEM-1, blaCTX-M-15, blaKPC-2 and blaNDM-1. Analysis of variance (ANOVA) and cluster analysis compared the number of resistant isolates per collection point and showed that a single location was statistically different from the others with regard to resistance. Although there are still no criteria to determine the envi ronmental contamination by resistant bacteria the fact that they have been isolated from flies is an indication of a disseminated contamination. As such, these insects may be useful in monitoring programs of antibiotic resistance in non-hospital environments, where they could function as sentinels.