Ceratophrys ornata is an iconic and regionally threatened amphibian of the temperate grasslands from Argentina, Brazil, and Uruguay. Conservation assessments suggested that habitat loss is the main threat to the species, although no studies have yet explored the link between grassland replacement and C. ornata decline. Using a robust database with ∼1000 records from herpetological collections, citizen science, and long-term monitoring, we modeled the species’ ecological niche to produce a map of suitable areas, where we searched for C. ornata using active and passive techniques. After exhaustive efforts, we failed to obtain recent records from Brazil and Uruguay. We also conducted Generalized Linear Models to explore the relationship between land use/cover, landscape configuration, and current species occurrence. Overall, results showed that C. ornata occurrence is negatively associated with variables related to native grasslands fragmentation and replacement (i.e., Distance to the Nearest Patch, Forest Plantations), suggesting that habitat loss may have driven local extinctions and population declines of the species. Finally, we outline the regional and national conservation needs of C. ornata and recommend focusing on in-situ conservation strategies for the Argentinean populations to ensure the species’ viability.