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Deoxyribonucleic acid (DNA) barcode (cytochrome c oxidase subunit I [COI]) sequences have been used to associate adult and immature stages of stoneflies as a rapid and effective alternative to rearing immatures. However, no molecular associations have been made for Neotropical species, which have very few described immature stages. Kempnyia Klapálek (Perlidae) is an endemic South American genus. Currently, 20 females have been associated with males of the 40 valid species, but only 5 nymphs have been described. We collected stonefly nymphs at Macaé River Basin, Rio de Janeiro State, Brazil, and Kempnyia nymphs were separated those in 6 morphotypes. These morphotypes were associated to nominal Kempnyia species using 38 COI sequences (471 base pairs [bp]) of immatures and identified adults of 10 species occurring in southern and southeastern Brazil. Based on a neighbor-joining …