We report the roadkill of a jaguar in one of the longest highways in Brazil (BR-101), in the stretch where this road crosses one of the most important Atlantic Forest remnants in the country: the Linhares-Sooretama block. The jaguar population present in this area represents the very last in entire Espírito Santo state. There is an approved project to the lines duplication of the entire BR-101 Highway and the company responsible by the work has already started the first activities in the state. However, there is no environmental impact assessment already done neither planning for the implementation of measures to avoid or reduce the roadkill risk in the region. Thus, to minimize the impacts associated with the BR-101, we do not recommend its lines duplication along the 15 km stretch traversing the Linhares-Sooretama block. In addition, alternatively, we suggest the deviation of the current route of the BR-101 Highway or the construction of overpasses to fauna in the most critical points, interspersing these overpasses with electronic speed monitoring devices and warning and educational plates.