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Three new cave-dwelling aeglid species from the Alto Ribeira karst region, south-eastern Brazil, are described. Morphological descriptions are based on traits traditionally used in aeglid taxonomy. Molecular phylogeny reconstruction based on the partial fragments of the 16S ribosomal RNA (16S rDNA) and the cytochrome oxidase subunit I (COI) revealed that: (1) dispersal of epigean aeglids toward the Alto Ribeira karst area occurred at two separate events in the evolutionary history of freshwater aeglids; (2) extant cave-dwelling aeglid species form two distinct subclades, each of which originated from a distinct ancestral epigean lineage; and (3) successful adaptation to subterranean habitats, as seen in extant cave-dwelling aeglid species from the Alto Ribeira karst area, developed independently within each subclade, as each one of them comprised an exclusive group of both stygobitic and stygophilic species. ZooBank: Three new cave-dwelling aeglid species are described from the Alto Ribeira karst region, south-eastern Brazil using morphological and molecular (16S rDNA) data. Comparison of molecular sequences from several subterranean and epigean species in the region reveal two dispersal events involving epigean ancestors that resulted in two distinct subclades of extant cave-dwelling species. Adaptation to subterranean habitats among the cave-dwelling species developed independently within each subclade resulting in each having an exclusive group of both stygobitic and stygophilic species. (Image credit: Sérgio Bueno.) [ABSTRACT FROM AUTHOR]