Eimeria separata: method for the excystation of sporozoites

A method is described for the excystation and collection of infective sporo zoites of Eimeria separata. The procedure uses conditions that resemble the in vivo environment. The first treatment of the oocysts in a 0.4% pepsin/H Cl solution alters the oocyst wall, which becomes thinner. The second treat ment in a 0.4% trypsin/0.75% taurocholate solution breaks the oocyst wall a nd sporocysts are released. A third incubation of the oocyst-sporocyst mixt ure in trypsin-free medium with 0.75% taurocholate and an additive of MgCl2 followed by a final incubation in RPMI medium supplemented with 1% fetal c alf serum yields a sporozoite excystation rate of up to 90%.