Submitted to: Journal of Parasitology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 10/17/2006
Publication Date: 6/1/2007
Citation: Obrien, C.N., Jenkins, M.C. 2007. A rapid method for production of high numbers of purified Cryptosporidium parvum oocysts. Journal of Parasitology. 93:434-436. Interpretive Summary: Cryptosporidiosis is an intestinal disease of humans and animals caused by the protozoan parasite Cryptosporidium parvum. The disease is prevalent in the young because of an incompletely developed immune system. Because of this lack of immunity, young calves are used to propagate Cryptosporidium oocysts. Large numbers of purified oocysts are necessary to study the proteins of the parasite in order to understand enzymatic processes and develop diagnostic tools. Purifying oocysts from fecal material can be a time consuming, expensive process resulting in low yields and purity. This rapid procedure combines a sodium chloride flotation step followed by an ether extraction to recover a large number of Cryptosporidium oocysts devoid of contaminating fecal debris or bacteria. The high yield of the oocyst preparation minimizes the number of propagations necessary and the high purity ensures the protein assays are devoid of microbe contamination.
Technical Abstract: Most procedures that have been described for purifying Cryptosporidium parvum oocysts are designed to either identify the parasites in clinical specimens or isolate oocysts from a small volume of feces from infected animals. The present study describes a rapid method for purifying high numbers of C. parvum oocysts from feces of infected calves that are virtually free of contaminating bacteria and yeast. This procedure regularly yields > 109 purified C. parvum oocysts within 1-2 days of feces collection.