Normal gametocyte numbers and sex ratio. Mean male gamete exflagellation events were slightly but significantly reduced in the psop25 line, as compared to the WT parasites.

Significantly reduced ookinete production (60% in vitro). Mature ookinetes formed showed normal (light microscope) morphology.

Significantly reduced oocyst production (69%)

More than 20-fold reduction in oocyst numbers.The size of parasite oocysts was clearly smaller than that of the wild-type parasite oocysts [as observed by phase contrast microscopy at 14 days post-infection (dpi). To identify the step in which they decreased in number, parasites were generated that constitutively expressed GFP from these disruptant populations and the oocysts shortly after ookinete invasion of the midgut at 2 dpi were counted. The number of oocysts was already approximately 10-fold smaller than that of wild-type parasites.We further measured oocyst diameters by epifluorescence microscopy at 14 dpi. The average oocyst diameter was approximately 60% of that of the wild-type oocysts, suggesting that oocyst development was impaired by this disruption.

In Anopheles coluzzi no oocyst formation

In Anopheles coluzzi no oocyst formation

Strongly reduced sporozoite formation

In Anopheles coluzzi no oocyst formation; no sporozoite formation (in A stephensi low numbers of oocysts were observed, but no sporozoites).

In Anopheles coluzzi no oocyst formation; no sporozoite formation (in A stephensi low numbers of oocysts were observed, but no sporozoites).