AbstractRoads pose an imminent threat to wildlife directly through mortality and changes in individual behavior, and also indirectly through modification of the amount and configuration of wildlife habitat. However, few studies have addressed how these mechanisms interact to determine species response to roads. We used structural equation modeling to assess direct and indirect effects (via landscape modification) of roads on space use by jaguars in Brazil, using radio-tracking data available from the literature. We fit path models that directly link jaguars’ space use to roads and to land cover, and indirectly link jaguars’ space use to roads through the same land cover categories. Our findings show that space use by jaguars was not directly affected by roads, but indirect effects occurred through reductions in natural areas on which jaguars depend, and through urban sprawl. Males´ space use, however, was not negatively influenced by urban areas. Since jaguars seem to ignore roads, mitigation should be directed to road fencing and promoting safe crossings. We argue that planners and managers need to much more seriously take into account the deforestation and the unbridled urban expansion from roads to ensure jaguar conservation in Brazil.