The exact taxonomic identity of members of the Simulium damnosum Theobald complex in Ethiopia was not known until recently. A cytotaxonomic analysis of larval chromosomes from the Kulfo River area in southern Ethiopia, from where the ‘Kulfo’ form was reported, and from the Awash River at Sodere in central Ethiopia has revealed the existence of two newly recognized species, namely Simulium kulfoense and Simulium soderense. Although three fixed and 11 floating inversions in the chromosomes of the ‘Kulfo’ form were described in an idiogram, in the present study four fixed and 17 floating inversions were observed, out of which only seven floating inversions were the same as Dunbar's, indicating that the remaining 10 inversions were either missed or misinterpreted. Though both species share many inversions, S. kulfoense sp. nov. has a diagnostic inversion on chromosome II, IIL-E6, with no sex-linked chromosomal arrangement, while S. soderense sp. nov. has a sex-linked inversion, IIS-3, which warrants it as a distinct species. Both of these new species also share many inversions with the recently described new species from south-western Ethiopia, Simulium kaffaense, including the fixed inversions IIL-E1 and E3. Since both the study sites are known to be free of onchocerciasis, and the flies are not anthropophilic, both the species are not presumed to be vectors of human onchocerciasis. The presence of IS-1 and a complex inversion possibly involving IL-3 strongly indicates that the two species belong to the ‘Nile’ phylogenetic group of S. damnosum s.l.