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Ecological interactions are being affected at unprecedented rates by human activities in tropical forests. Yet, the continuity of ecological functions provided by animals, such as seed dispersal, is crucial for forest regeneration and species resilience to anthropogenic pressures. The construction of new roads in tropical forests is one of the main boosters of habitat destruction as it facilitates human access to previously isolated areas and increases defaunation and loss of ecological functions. It, therefore, becomes increasingly urgent to rapidly assess how recently opened roads and associated anthropogenic activities affect ecological processes in natural habitats, so that appropriate management measures to conserve diversity can be taken. In this study, we aimed to evaluate the effects of anthropogenic pressures on the health status of a mature rainforest crossed by a newly opened road in French Guiana. For this, we combined different methods to conduct a rapid assessment of the forest’s health status. Firstly, we evaluated the activity of frugivores using camera traps deployed in four forest patches located near (