The genus Lytopylus Foerster, 1862 (Hymenoptera: Braconidae: Agathidinae) has 40 described species with a primarily Neotropical distribution. The present work presents three new species of the genus: Lytopylus melanoflavus Silva, Shimbori & Fernandes sp. nov. and Lytopylus tigrinus Silva, Shimbori & Fernandes sp. nov. from the Amazon rainforest (State of Amazonas), and Lytopylus minimus Silva, Shimbori & Fernandes sp. nov. from the Cerrado (State of Mato Grosso). Additionally, we provide an identification key for all Brazilian species of Lytopylus.