Anopheles fluviatilis James is an important malaria vector in Indian subcontinent. An. fluviatilis exists as a complex of three sibling species, of which two species, T and U, have been colonized so far. Attempts were made to study the comparative susceptibility of species T and U of the An. fluviatilis complex to rodent malaria parasite Plasmodium vinckei petteri by using Anopheles stephensi Liston as calibrator for variable infectivity in different isolates. An. stephensi, which was used as control, became readily infected, with 60–65% mosquitoes carrying developing oocysts, whereas in species T and species U, ≈50 and 63%, respectively, of mosquitoes carried oocyts. An. fluviatilis species T was found comparatively less susceptible to P. v. petteri sporogonic development compared with species U. Moreover, significantly lesser sporozoites rate (11%) was observed in species T compared with 31% in species U. Species T and species U are not considered as malaria vectors in India in the field. However, in the laboratory, both these species are able to support the malaria sporogony.