The Neotropical species of Asthenopodinae are revised in a formal phylogenetic context. The five known species of Asthenopus Eaton, 1871, together with other five new species were included in a cladistic analysis using morphological characters (continuous and discretes). Representatives of the Afro-Oriental group of the subfamily (Povilla Navás, 1912 and Languidipes Hubbard, 1984) were also included to test the monophyletic hypothesis traditionally accepted for the group. Additional taxa representing the other subfamilies of Polymitarcyidae were incorparated: Ephoron Williamson, 1802 (Polymitarcyinae) and Campsurus Eaton, 1868, Tortopus Needham & Murphy, 1924 and Tortopsis Molineri, 2010 (Campsurinae). A matrix of 17 taxa and 72 characters was analyzed under parsimony resulting in a single tree supporting the monophyly of the subfamily Asthenopodinae. Other results include the monophyly of the Afro-Oriental taxa (Povilla and Languidipes), the paraphyletic nature of Neotropical Asthenopodinae, and the recognition of four South American genera: Asthenopus (including Asthenopus
curtus (Hagen), 1861, Asthenopus
angelae de Souza & Molineri, 2012, Asthenopus
magnus
sp. n., Asthenopus
hubbardi
sp. n., Asthenopus
guarani
sp. n.), Asthenopodes Ulmer, 1924, stat. n. (including Asthenopus
picteti Hubbard, 1975, stat. n., Asthenopodes
traverae
sp. n., Asthenopodes
chumuco
sp. n.), Priasthenopus
gen. n. (including Priasthenopus
gilliesi (Domínguez), 1988, comb. n.), and Hubbardipes
gen. n. (including Hubbardipes
crenulatus (Molineri et al.), 2011, comb. n.). Descriptions, diagnoses, illustrations and keys are presented for all Neotropical taxa of Asthenopodinae (adults of both sexes, eggs and nymphs). Additionally a key to the subfamilies and genera of Polymitarcyidae is included. A quantitative biogeographic analysis of vicariance is presented and discussed through the study of the “taxon history” of the group.