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Passive forest restoration can buffer the effects of habitat loss on biodiversity. We acoustically surveyed aerial insectivorous bats in a whole‐ecosystem fragmentation experiment in the Brazilian Amazon over a 2‐year period, across 33 sites, comprising continuous old‐growth forest, remnant fragments, and regenerating secondary forest matrix. We analyzed the activity of 10 species/sonotypes to investigate occupancy across habitat types and responses to fragment size and interior‐edge‐matrix (IEM) disturbance gradients. Employing a multiscale approach, we investigated guild (edge foragers, forest specialists, flexible forest foragers, and open space specialists) and species‐level responses to vegetation structure and forest cover, edge, and patch density across six spatial scales (0.5–3 km). We found species‐specific habitat occupancy patterns and nuanced responses to fragment size and the IEM disturbance …