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Background  Taxonomic identification of triatomines is generally performed based on aspects of their external morphology. However, the use of a multidisciplinary approach, considering morphological aspects of the external genitalia, morphometry, genetics, and phylogeography has been suggested, especially for similar and/or cryptic species. The rupestral species  Triatoma jatai  Gonçalves et al., 2013,  Triatoma costalimai  Verano & Galvão, 1959 and  Triatoma williami  Galvão et al., 1965, which are morphologically similar, have been found naturally infected with  Trypanosoma cruzi  (Chagas, 1909) in wild, peridomestic, and intradomestic environments, representing a risk of new outbreaks of Chagas disease. This study presents morphological description complementation of these species, with an emphasis on the structures of the female external genitalia, using scanning electron microscopy.    Methods  The females of  T. jatai  and  T. costalimai  ( n  = 10 of each) were captured in the Brazilian municipalities of Paranã and Aurora do Tocantins and were identified with the use of a dichotomous key for the Matogrossensis subcomplex. Females of  T. williami  ( n  = 5), were obtained from a laboratory colony. The females were cut transversely at the sixth abdominal segment and examined under scanning electron microscopy (SEM) at the Oswaldo Cruz/Fiocruz Institute Electronic Microscopy Platform.    Results  It was possible to differentiate the three species based on the characteristics of urotergites VII, VIII and IX and urosternite VII, as well as the genital plaques, gonocoxites, and gonapophyses. To our knowledge, morphological differences in the spines present on gonapophysis 8 in triatomines are described here for the first time.    Conclusions  The results show that external genitalia of females are useful structures to differentiate  T. costalimai ,  T. jatai  and  T. williami . SEM analysis contributes to and corroborates, together with other tools morphological and molecular, the distinction of the three species.

External female genitalia of Triatoma jatai, Triatoma costalimai and Triatoma williami (Hemiptera: Reduviidae: Triatominae)