Anopheles Pupa Collection and Sex Identification

For most Anopheles species, larval-pupal metamorphosis commences ∼1 wk after egg hatching. However, depending on the amount of food provided, H₂O temperature, and larval density, the pupation process can be accelerated or delayed. Synchronous pupation is difficult to accomplish consistently, and, thus, pupae need to be separated from larvae daily. Adult emergence will take place 24-48 h after pupation. Most adults will eclose before the next morning (light cycle) in many species. Here, we provide information on some methods available to collect pupae and to sort pupae by sex. Notably, pupa collection and sorting are some of the most time-consuming procedures of the overall mosquito rearing process. Some methods mentioned here attempt to help reduce work effort and time required.