The revision of the Afrotropical Ophioninae is updated, based on the examination of about 800–900 individuals in the South African and European museum collections. A robust interactive matrix key was built to provide quick and reliable identifications. The key is available online at http://www.waspweb.org. Two new species are described: Dicamptus maxipol sp. n. and Enicospilus gauldetmitchellorum sp. n. Numerous new distribution and biological records are provided, and noticeable morphological intra-specific variations are detailed. Enicospilus batus Gauld & Mitchell, syn. n. is considered as a junior synonym of Enicospilus luebberti (Enderlein).