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Vegetation clutter is a limiting factor for bats that forage near ground level, and may determine the distribution of species and guilds. However, many studies that evaluated the effects of vegetation clutter on bats have used qualitative descriptions rather than direct measurements of vegetation density. Moreover, few studies have evaluated the effect of vegetation clutter on a regional scale. Here, we evaluate the influence of the physical obstruction of vegetation on phyllostomid-bat assemblages along a 520 km transect in continuous Amazonian forest. We sampled bats using mist nets in eight localities during 80 nights (3840 net-hours) and estimated the ground-vegetation density with digital photographs. The total number of species, number of animalivorous species, total number of frugivorous species, number of understory frugivorous species, and abundance of canopy frugivorous bats were negatively associated with vegetation clutter. The bat assemblages showed a nested structure in relation to degree of clutter, with animalivorous and understory frugivorous bats distributed throughout the vegetation-clutter gradient, while canopy frugivores were restricted to sites with more open vegetation. The species distribution along the gradient of vegetation clutter was not closely associated with wing morphology, but aspect ratio and wing load differed between frugivores and animalivores. Vegetation structure plays an important role in structuring assemblages of the bats at the regional scale by increasing beta diversity between sites. Differences in foraging strategy and diet of the guilds seem to have contributed more to the spatial distribution of bats …