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.

Roads impact wildlife around the world; however, dedicated studies are lacking in many biodiverse areas such as the Amazon. Identifying which species are more often hit by vehicles and which landscape and road-related features promote roadkill is essential to guide future development and ensure adequate mitigation actions. For six months, we monitored 240 km of roads in the Ecuadorian Amazon and recorded 1125 dead vertebrates (148 species). Reptiles were the most affected class with 380 individuals (56 species), followed by amphibians with 278 individuals (11 species), birds with 259 individuals (62 species), and mammals with 208 individuals (20 species). We used Random Forest models to explore the role of various land cover types and road sinuosity on the observed mortality. Additionally, we created heatmaps to visualize the road segments where roadkills were more frequent. For all vertebrates, mortality was more likely in straight road sections near rivers. The effect of other variables was taxa-specific. Amphibian mortality was more likely near bare soil or forest, birds and mammals died more often near herbaceous-shrubby vegetation, whereas reptile mortality occurred more often further from herbaceous-shrubby vegetation. Road segments with a high mortality (roadkill hotspots) varied across taxa. These hotspots identify areas where further research is needed to assess road impacts and where mitigation could prevent collisions. Among records, we found rare and threatened species, including some that may be new to science. Roadkill surveys not only aid in quantifying threats and informing future planning but can also provide insight into local biodiversity.

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