Laboratory culture of <i>Triatoma sanguisuga</i> (Hemiptera: Reduviidae) by supplementation with <i>Drosophila</i> artificial hemolymph media

Triatoma sanguisuga (LeConte, 1855), the eastern bloodsucking conenose, is an epidemiologically significant triatomine vector in the United States in that it is implicated in documented autochthonous human Chagas disease cases in the United States and can be found across a large geographic area. Despite decades of research interest, this species has proven remarkably challenging to maintain in laboratory culture, with consistent developmental arrest occurring prior to the fifth nymphal instar stage when maintained on standard blood-only feeding protocols. Here, we report a successful protocol for maintaining T. sanguisuga through complete development in laboratory culture. Adult specimens were collected from Des Allemands, Louisiana, and maintained under standard laboratory conditions with blood feeding supplemented by Drosophila artificial hemolymph media (AHL). Early nymphal development proceeded through the third instar on blood meals alone, but development consistently stalled at the fourth instar. Following initiation of AHL supplementation, the first successful fourth-to-fifth instar molt was observed approximately 3 wk later, with subsequent emergence of adult insects occurring within 17 wk. The AHL supplement was readily consumed by nymphs when offered ad libitum at room temperature. This development overcomes a significant barrier to T. sanguisuga laboratory culture and provides compelling evidence for complex nutritional requirements in triatomines beyond those met by vertebrate blood alone. Laboratory-reared T. sanguisuga cultures will enable critical research on vector competence, feeding behavior, reproductive biology, and control measures essential for understanding and managing Chagas disease transmission in North America.