Spilonympha, a new subgenus of the genus Wyeomyia Theobald, is described based on morphological characters of the adult, larval, and pupal stages. The taxonomic species included in the new subgenus are Wyeomyia mystes (Dyar), Wy. bourrouli (Lutz), Wy. forcipenis Lourenço-de-Oliveira & Silva, Wy. finlayi Lane & Cerqueira, Wy. airosai Lane & Cerqueira, Wy. howardi Lane & Cerqueira, and a new species designated Wy. aningae, which is described and illustrated. Wy. aningae was designated as the type species of the new subgenus. The systematic treatment of these species includes a key for the identification of the adult males, pupae, and larvae; bionomics data; and geographical distribution.