Vehicle collisions with wildlife have the potential to negatively impact species everywhere roads divide natural habitats; this is especially true in areas where roads are a recent addition to the landscape (e.g., in the last decade) and/or where expansion of a road network is underway. This study addresses the impact of traffic fatalities on lava lizards, Microlophus albemarlensis, on the island of Santa Cruz, Galápagos. The main road bisecting the island north to south was added to the island beginning in 1974 but not fully paved until 2000. We assessed impacts occurring due to traffic and vehicle speeds on the island. This study adds impacts to lizard populations to those conducted on birds on Santa Cruz, providing a means to create multi-species recommendations. To quantify impacts to lizard populations, we assessed lizards on randomly chosen transects perpendicular to the road. We used Poisson regression to analyze those data and found a 30% increase in lizard abundance per 100 m distance from the road across vegetation zones. We compared incidence of prior tail loss of road-killed lizards and live animals to comment on additional energetic costs of occupying near-road territories. Tail condition could not be assessed in some road-killed lizards; among those that we could assess, 29% showed evidence of prior tail loss. That rate declined to apparent baseline conditions of only 1% beyond 200 m from the road. By traveling the length of the 40-km road, we identified hot spots for fatalities of both lizards and birds to provide necessary information for developing management and remediation strategies. We offer guidance on Best Management Practices, such as testing overpasses, fencing and underpasses, which might reduce vehicle impacts to lizards from drivers on the current road. This information is essential because of a planned increase in the Santa Cruz road network in areas where lava lizards are most abundant.