Ecuador has both high richness and high endemism, which are increasingly threatened by anthropic pressures, including roads. Research evaluating the effects of roads remains scarce, making it difficult to develop mitigation plans. Here, we present the first national assessment of wildlife mortality on roads that allow us to (1) estimate roadkill rates per species, (2) identify affected species and areas, and (3) reveal knowledge gaps. We bring together data from systematic surveys and citizen science efforts to present a dataset with 5010 wildlife roadkill records from 392 species, and we also provide 333 standardized corrected roadkill rates calculated on 242 species. Systematic surveys were reported by ten studies from five Ecuadorian provinces, revealing 242 species with corrected roadkill rates ranging from 0.03 to 171.72 ind./km/year. The highest rates were for the yellow warbler Setophaga petechia in Galapagos (171.72 ind./km/year), the cane toad Rhinella marina in Manabi (110.70 ind./km/year), and the Galapagos lava lizard Microlophus albemarlensis (47.17 ind./km/year). Citizen science and other nonsystematic monitoring provided 1705 roadkill records representing all 24 provinces in Ecuador and 262 identified species. The common opossum Didelphis marsupialis, the Andean white-eared opossum Didelphis pernigra, and the yellow warbler Setophaga petechia were more commonly reported (250, 104, and 81 individuals, respectively). Across all sources, we found 15 species listed as "Threatened" and six as "Data Deficient" by the IUCN. We recommend stronger research efforts in areas where the mortality of endemic or threatened species could be critical for populations, such as in Galapagos. This first country-wide assessment of wildlife mortality on Ecuadorian roads represents contributions from academia, members of the public, and government, underlining the value of wider engagement and collaboration. We hope these findings and the compiled dataset will guide sensible driving and sustainable planning of infrastructure in Ecuador and, ultimately, contribute to reduce wildlife mortality on roads.