We used roost searches, mist netting and acoustic sampling to investigate the habitats used by bats in Parc National de Mantadia and the Réserve Spéciale d'Analamazaotra, eastern Madagascar. Four species were caught in relatively intact humid forest (Myotis goudoti, Miniopterus manavi, Miniopterus majori and Emballonura atrata) two in agricultural land, Neoromicia matroka and Neoromicia melckorum, and one, Rousettus madagascariensis, in Eucalyptus plantations. Mormopterus jugularis, Chaerephon pumilus and Mops leucostigma were found roosting in buildings ca. three km from the humid forest. Acoustic sampling revealed that Neoromicia spp. and molossids were ubiquitous and were recorded from intact and degraded humid forest, Eucalyptus plantations and agricultural land. Myotis goudoti showed the strongest association with intact humid forest. Taxon richness, determined by acoustic sampling, was highest in humid forest but activity was highest in plantations and agricultural land. Mixed-habitat landscapes that surround protected forests and consist of a mosaic of regenerating forest, agriculture, wetlands, villages and plantations are important for bats and promote chiropteran diversity because they provide roosting and foraging sites for species that rarely use intact forest. The humid forests of eastern Madagascar have lower bat diversity than the island's western deciduous karst forests.