Studies on Plasmodium ookinetes. 1. Isolation and concentration from mosquito midguts

In a method for isolating a relatively clean suspension of concentrated Plasmodium berghei ookinetes from infected midguts of Anopheles stephensi at appropriate times after the infective blood meal, the ookinetes are freed from the midguts by enzymatic digestion, and then concentrated by means of a BSA/renografin gradient. The mean number of ookinetes recovered/midgut was 152. More than 95% of the recovered ookinetes were viable by the criteria of motility, incorporation of adenosine and leucine, and appearance on light and electron microscopic examination. Trypan blue exclusion was not a valid criterion for viability. These ookinetes were not useful for in vitro studies of further development due to the presence of contaminating microorganisms. Our attempts to determine their potential for further development in vivo have similarly not been successful. Nevertheless, our ability to obtain large numbers of ookinetes at defined times during development now permits further studies on their structure, biochemistry, and physiology, as well as comparison with ookinetes formed in vitro.