The infectivity and purification of cultured Plasmodium berghei ookinetes

Abstract

Plasmodium berghei ookinetes were cultured from hamster blood as described previously (Kurtti and Munderloh, 1986). An average of 7.3 X 10(6) ookinetes was harvested from each ml of blood. Ookinetes were purified by centrifugation on first a 40% and then a 36% Percoll gradient. The final preparation comprised 32.8% of the ookinetes initially obtained, and contained 3.3 other parasite stages or blood cells per ookinete. Unpurified and purified ookinetes were resuspended in hamster blood and fed to Anopheles stephensi. There was a strong linear correlation between the concentration of purified or unpurified ookinetes and the number of oocysts formed. With unpurified ookinetes, a maximum was reached when preparations containing 1 X 10(7) ookinetes/ml were fed, and feeding preparations containing a higher concentration did not produce more oocysts. Sporozoites were found in the salivary glands of mosquitoes fed ookinetes by days 14 (unpurified) or 15 (purified) PI. Approximately 5 times as many purified as unpurified ookinetes were required to produce each oocyst.