A climate model, based on effects of water availability and temperature, was recently proposed to explain global variation in bat species richness along altitudinal gradients. Yet such studies are sparse in the tropics and near-absent in Africa. Here we present results from an altitudinal study of bat diversity from Mount Mulanje, Malawi. Using ground nets, canopy nets and harp traps, we sampled eight sites across three habitat zones from 630 m to 2010 m asl. We assessed the influence of climatic, geographic and biotic variables on measures of estimated species richness, Fisher's α, and an unbiased index of compositional turnover. We recorded 723 individuals and 30 species along the gradient, revealing a ‘low plateau’ pattern in estimated species richness, peaking at 1220 m, which is congruent with the global climate model. Measures of local habitat structure significantly explained a large degree of variation in species richness and compositional turnover between sites. Fisher's α was further significantly correlated to mean annual relative humidity, suggesting a background climatic influence.