Protocol for assembling and implementing a partially automated system for rearing and handling Anopheles stephensi mosquitoes

Live mosquitoes are required to comprehensively study vector-borne diseases, including transmission. Traditional mosquito-rearing protocols are laborious and time consuming. Here, we present a protocol for assembling and implementing a partially automated system for rearing and handling Anopheles stephensi mosquitoes. We describe steps for assembling a pupation station, self-emptying bucket, pupal funnel and dish vacuum, automatic aspirator, and sugar tubes. We also detail the application of these systems, along with specific limitations.