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Title: Obtaining Highly Purified Toxoplasma gondii Oocysts by a Discontinuous Cesium Chloride Gradient

Author
item STAGGS, SARAH - Dynamac Corporation
item SEE, MARY - University Of Cincinnati
item Dubey, Jitender
item VILLEGAS, ERIC - Environmental Protection Agency (EPA)

Submitted to: Journal of Visualized Experiments
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/30/2009
Publication Date: 11/3/2009
Citation: Staggs, S.E., See, M.J., Dubey, J.P., Villegas, E.N. 2009. Obtaining Highly Purified Toxoplasma gondii Oocysts by a Discontinuous Cesium Chloride Gradient. Journal of Visualized Experiments. http://dx.doi.org/10.3791/1420.

Interpretive Summary: Toxoplasma gondii is a single-celled parasite of all warm-blooded hosts worldwide. It causes mental retardation and loss of vision in children, and abortion in livestock. Cats are the main reservoir of T. gondii because they are the only hosts that can excrete the resistant stage (oocyst) of the parasite in the feces. Humans become infected by eating undercooked meat from infected animals and food and water contaminated with oocysts. In the present paper, scientists at the Beltsville Agricultural Research Center and Us Environmental Protection Agency report a method to separatye T. gondii oocysts from cat feces. The results will be of interest to biologists, parasitologists, and veterinarians.

Technical Abstract: Toxoplasma gondii is an obligate intracellular protozoan pathogen that commonly infects humans. To date, research on understanding the prevalence of T. gondii oocysts in the water and environment are limited due to the lack of tools to detect oocysts in the environment. This is primarily due to the lack of efficient purification protocols for obtaining large numbers of highly purified T gondii oocysts from infected cats for research purposes. This study describes the development of a modified CsCl method that easily purifies T. gondii oocysts from feces of infected cats that are suitable for molecular biological and tissue culture manipulation.