Development of malaria parasites in mosquitoes fed with ookinetes suspended in defined media

Information concerning the specific nutritional requirements of malarial parasites developing in the mosquito host has been difficult to obtain, owing primarily to the complex nature of the blood meal that accompanies the parasites and the lack of success in culturing the complete invertebrate cycle of Plasmodium in vitro. The present report describes a blood-free system for infecting mosquitoes with ookinetes of Plasmodium berghei and for allowing the latter to develop into infective sporozoites. Ookinetes cultured in vitro were separated from blood proteins, suspended in defined medium, and fed to Anopheles stephensi mosquitoes through a membrane. The mosquitoes were then maintained on the same defined medium plus 5% sucrose. Infectivity of the parasites was demonstrated 17-19 days later by intracardial inoculation of the macerated mosquitoes into hamsters. This system makes it possible to evaluate nutritional factors that affect parasite development in the mosquito host under controlled conditions.