Production of Plasmodium falciparum oocysts and sporozoites

The production of different sexual stages of P. falciparum and their isolation from infected mosquitoes facilitates a variety of cellular, molecular, immunochemical, and transmission-blocking studies that examine the parasite-vector relationship. The procedures of mosquito rearing, parasite cultivation, and infection and dissection of mosquitoes are highly specialized skills requiring considerable experience to perform with competence. Additionally, the timing of the two life cycles, that of the parasite and that of the mosquito, requires precise planning so that the mosquitoes are the correct age to infect during the brief period in which the parasites are viable for infection. Mosquitoes capable of transmitting P. falciparum should be maintained within a limited access, dedicated, high security humidified insectary. It is recommended that all rooms be painted in a light color and all cages be constructed of white materials in order to make highly visible any mosquito that may temporarily escape. A nonresidual insecticide spray should also be available in the event of an emergency. Infected mosquitoes to be dissected are first stunned or anesthetized by chloroform and then rendered incapable of movement (dewinged and delegged). Any potentially infective mosquitoes that are not to be dissected are frozen by placing the complete container at minus 20 degrees celsius prior to autoclaving and disposal.