Protocol for laboratory rearing and infection tracking of Rhodnius prolixus using 3D-printable designs

Human infections by Trypanosoma cruzi propagate via its blood-feeding triatomine vector. Investigating parasite-vector interactions depends upon robust techniques to rear insects and analyze infections. Here, we present a protocol for laboratory rearing and infection tracking of Rhodnius prolixus. We describe steps for housing, feeding, and sorting strategies using 3D-printable designs. We also detail procedures for gut dissection, fecal collection, and parasite re-isolation. This protocol describes techniques that support efforts to understand and mitigate vector-mediated Chagas disease transmission.