The purpose of this study was to evaluate the use of rhodamine B as an orally delivered biomarker for rodents and a feed-through transtadial biomarker for phlebotomine sand flies (Diptera: Psychodidae ). Rhodamine B-treated hamsters were visibly marked for up to 8 wk, and their feces were fluorescent when examined under a fluorescence microscope. The development and survival of sand fly larvae fed feces of rhodamine B-treated hamsters were not significantly different from control sand flies. Adult male and female sand flies, that had been fed as larvae the feces of rhodamine B-treated hamsters, were fluorescent when examined using fluorescent microscopy and could be distinguished from control sand flies. Adult female sand flies that took bloodmeals from rhodamine B-treated hamsters were fluorescent when examined immediately after feeding. Rhodamine B incorporated rodent baits could be used to detect adult male and female sand flies that fed on the feces of baited rodents as larvae, or adult female sand flies that have taken a bloodmeal from bait-fed rodents. This would allow the delineation of specific foci with rodent-sand fly associations that would be susceptible to control by using feed-through or systemic insecticides.