ABSTRACT. Diversity of Ephemeroptera (Insecta) of the Serra da Mantiqueira and Serra do Mar, southeastern Brazil. The aim of this study was to inventory the mayfly fauna, based on nymphal and alate stages, in Serra da Mantiqueira and in Serra do Mar, São Paulo State, as well as to present information about habitats used by the genera. Nymphs were collected in several streams and mesohabitats with a Surber sampler and the winged stages with light attraction methods, entomological nets, and Malaise traps. In all, eight families and 33 genera were recorded, representing a very significant portion of the Brazilian fauna (80% of families and 49% of genera). Furthermore, it was possible to identify 11 species, of which two are new records for the state: Tricorythodes santarita Traver and Caenis reissi Malzacher. Despite the high diversity recorded, the accumulation curves presented an ascending form, indicating an increase in the number of genera with additional sampling effort. The high richness found in these areas are in agreement with the high biodiversity of the Atlantic Forest biome and the sampling effort employed, which included the use of different methods, the collection of both nymphs and winged stages, and the sampling of a large area with diverse streams and habitats.