Studies about processes determining biodiversity in Pantanal are urgently needed and the use of invertebrates for this purpose may be an efficient tool. The purpose of this paper is to verify whether ants could be used to predict human impacts on Pantanal, assessing how environmental characteristics of gallery forest influence arboreal ant species richness. Specifically, we tested the hypotheses that: (i) resource availability (estimated by tree density) and (ii) diversity of environmental conditions (estimated by structural heterogeneity) positively influence arboreal ant species richness and composition. Ants were sampled in a gallery forest in the Miranda River. Ant species richness did not respond to tree density, but increased with structural heterogeneity. Species composition was affected both by tree density and structural heterogeneity. The species list produced in the present study showed a high similarity between arboreal ant faunas of Cerrado and Pantanal, which are contiguous biomes. The relationship of ant species richness with heterogeneity, as well as the observed changes of species composition with tree density and heterogeneity may be explained by the presence of a variation of environmental conditions, which create several micro-habitats in the forest. The response of the ant community can reflect human impacts on the forests of the studied region, which are usually modified to create patches for sport fishing and by pathways used in ecotourism.