Understanding carnivores' diet is key to understanding their adaptability in a rapidly changing world. However, studying diet of large carnivores is difficult due to their elusive nature. In this study, we performed DNA metabarcoding analyses of 82 putative leopard scats collected from two distinct, but connected, habitat types (rainforest and grassland) in the Udzungwa Mountains, Tanzania. Two mitochondrial markers were used to identify predator and prey. Metabarcoding confirmed that 60 of the collected scats (73%) originated from leopards, and nineteen mammalian prey DNA sequences were identified to species. Using prey size correction factors for leopards, and covariates on habitat type and prey ecology, we investigated whether differences in leopard dietary composition were detectable between habitats. We found that leopards in grassland consumed a larger mean prey size compared with leopards in rainforest. Small prey (70% of the biomass consumed by leopards in rainforest, while large prey (>= 80 kg) were only eaten in grassland. Arboreal species constituted 50% of the biomass consumed by rainforest leopards. Our results highlight the importance of arboreal species in their diet. From a management perspective, we suggest continued protection of all prey species in the protected areas to prevent human-wildlife conflicts.