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Morphological and molecular sequence data were combined with cross-hybridization studies and used to identify a new Steinernema sp. from Free State, South Africa. Molecular and morphological data indicate that the new species belongs to the ‘glaseri-group’ of Steinernema spp. Key morphological diagnostic characters for S. innovationi n. sp. include the morphometric features of the third-stage infective juveniles: total body length = 1054 (1000–1103) μm, tail length = 108 (97–117) μm, location of the excretory pore = 88 (82–91) μm, and D% = 58 (54–63), E% = 115 (104–137) and H% = 43 (37–46). Additionally, the morphology of the spicules and gubernaculum of the first-generation males are considered key diagnostic traits. Steinernema innovationi n. sp. was also characterized by analysis of both rDNA and mitochondrial gene sequence data, which further indicate the uniqueness of this Steinernema species. [ABSTRACT FROM AUTHOR]