A new scorpion species, Tityus imei sp. nov., is described, representing the first record of the genus for the southern foothills of Sierra de Portuguesa in the western Venezuelan state of Portuguesa. The species is morphologically related to T. sanarensis, from the state of Lara, and T. boconoensis, from state of Trujillo. Tityus imei sp. nov. can be distinguished by the array of dorsal trichobothria in the pedipalp femur (d3, d4, d5) and by the following combination of female metasomal segment II characters: (i) ventral keels double and parallel up to two thirds of the segment, whereupon they converge into a single keel and separate again in the base; (ii) full, parallel ventral keels, including a well defined central keel in the second half of the segment. A molecular approach was undertaken to estimate the genetic relationship between the new species, T. sanarensis, T. boconoensis, and T. discrepans. Sequencing of a 562 bp-segment encoding sectors M6 to M11 of the cytochrome oxidase subunit I revealed that T. imei sp. nov. diverges from T. sanarensis and T. boconoensis, particularly in the polymorphic sector I4, sharing more overall sequence similarity with the northcentral Venezuelan species, T. discrepans. The new species is of potential medical importance given the presence of venom components whose mass resembles that of scorpion neurotoxins affecting voltage-sensitive sodium channels, as indicated by MALDI-TOF MS analyses.