Development of human and calf Cryptosporidium in chicken embryos

Cryptosporidium is a newly recognized, zoonotic protozoan that produces short-term, flu-like, gastrointestinal illness in immunocompetent humans and prolonged, severe, diarrhea in immunocompromised individuals. Successful completion of the life cycle, from sporozoite to infective oocyst, of isolates of Cryptosporidium from humans and calves was demonstrated in endoderm cells of the chorioallantoic membrane (CAM) of chicken embryos maintained at 37 C. The human and calf isolates of Cryptosporidium were morphologically and developmentally indistinguishable when grown in chicken embryos. The human isolate also completed its entire life cycle in the CAMs of chicken embryos maintained at 35 C and 41 C. Oocysts recovered from endoderm cells of infected CAMs produced heavy infections in suckling mice. The timing, presence, and morphology of developmental stages in CAM cells during the first eight days after inoculation of sporozoites were similar to those in enterocytes of mice inoculated with oocysts. The method described is safe and convenient for cultivating and studying Cryptosporidium in a bacteria-free environment; the system also lends itself to well-established procedures for evaluating antiprotozoan drugs.