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

## TL;DR

This paper provides a detailed protocol for rearing and tracking infections in Rhodnius prolixus, a vector for Chagas disease, using 3D-printable tools.

## Contribution

The novel contribution is a 3D-printable, cost-effective protocol for rearing Rhodnius prolixus and tracking Trypanosoma cruzi infections in the lab.

## Key findings

- A 3D-printable system for housing and feeding Rhodnius prolixus is described.
- Procedures for collecting and analyzing excreta from infected insects are detailed.
- A method for isolating Trypanosoma cruzi from the gut or excreta of R. prolixus is provided.

## Abstract

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.

•Assembly instructions for partially/fully disposable caging for Rhodnius prolixus•Design for artificial blood-feeding system using water balloons and handwarmers•Guidelines for collection of excreta from Trypanosoma cruzi-infected R. prolixus•Procedure to isolate T. cruzi from R. prolixus gut or excreta for axenic culture

Assembly instructions for partially/fully disposable caging for Rhodnius prolixus

Design for artificial blood-feeding system using water balloons and handwarmers

Guidelines for collection of excreta from Trypanosoma cruzi-infected R. prolixus

Procedure to isolate T. cruzi from R. prolixus gut or excreta for axenic culture

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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.