The aim of the present study was to explore the diversity and describe the spatial distribution of Calliphoridae and Sarcophagidae along an urbanization gradient in the Iberá Wetlands. The flies were collected at 18 sampling points, three per site. In total, 3,318 flies were collected (3,077 blow flies and 241 flesh flies), distributed in 13 genera and 33 species. Calliphoridae was the most abundant, comprised 92.74% of all adult flies. Chrysomya albiceps (Wiedemann), Cochliomyia macellaria (F.), Chrysomya megacephala (F.), and Chrysomya putoria (Wiedemann) were the most abundant species, representing 82.21% of the total sample. The genus Oxysarcodexia Townsend was the most diverse with 10 species represented. The abundance of exotic species represents 62.85% of the total sample, showing a reduction toward less disturbed sites. The results of this study indicated that sites with intermediate impact had higher diversity than those where the disturbances are high or absent. Our findings provide the first assessment of Calliphoridae and Sarcophagidae diversity in the area of the Iberá Wetlands.