Malarial parasites complete sporogony in axenic mosquitoes

To obtain sporogonic stages of malaria free from microbial contaminants for in vitro studies, Anopheles stephensi were reared under sterile conditions using a mosquito cell line as larval food. The adult females, kept in sterile humidified containers and allowed to engorge on parasitemic hamsters, supported the sporogonic development of the rodent malarial parasite Plasmodium berghei. In 10 experiments, the proportion of infected mosquitoes varied from 0 to 92%, and the geometric mean number of oocysts per female mosquito from 2.5 to 58.6, with a range of 1 to 548. The average number of salivary gland sporozoites per infected mosquito was determined by direct sporozoite counts in the pooled homogenate of the thoraces of all female mosquitoes. In five experiments, it varied from 2.7 X 10(3) to 9.0 X 10(3). The sterile sporozoites, harvested on day 19 or 20 after the infective blood meal, were as infective for rodents as nonsterile ones.