Abstract Tropical rainforests usually have multiple strata that results in a vertical stratification of ecological opportunities for animals. We investigated if this stratification influences the way bats use the vertical space in flooded and unflooded forests of the Central Amazon. Using mist-nets set in the canopy (17 to 35 m high) and in the understorey (0 to 3 m high) we sampled four sites in upland unflooded forests (terra firme), three in forests seasonally flooded by nutrient-rich water (várzea), and three in forests seasonally flooded by nutrient-poor water (igapó). Using rarefaction curves we found that species richness in the understorey and canopy were very similar. An ordination analysis clearly separated the bat assemblages of the canopy from those of the understorey in both flooded and unflooded habitats. Gleaning carnivores were clearly associated with the understorey, whereas frugivores were abundant in both strata. Of the frugivores, Carollinae and some Stenodermatinae were understorey specialists, but several Stenodermatinae mostly used the canopy. The first group mainly includes species that, in general, feed on fruits of understorey shrubs, whereas the second group feed on figs and other canopy fruits. We conclude that vertical stratification in bat communities occurs even within forests with lower canopy heights, such as Amazonian seasonally flooded forests, and that the vertical distribution of bat species is closely related to their diet and foraging behaviour.