Isolation, Culture and Cryopreservation of Sarcocystis species

Authors: VERMA, SHIV - Non ARS Employee; LINDSAY, DAVID - Virginia-Maryland Regional College Of Veterinary Medicine (VMRCVM); GRIGG, MICHAEL - National Instiute Of Allergy And Infectious Diseases (NIAID, NIH); Dubey, Jitender

Submitted to: Current Protocols in Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/1/2017
Publication Date: 5/1/2017
Citation: Verma, S., Lindsay, D., Grigg, M., Dubey, J.P. 2017. Isolation, Culture and Cryopreservation of Sarcocystis species. Current Protocols in Microbiology. doi: 10.1002/cpmc.32.

Interpretive Summary: Toxoplasma and Sarcocystis are related single celled parasites of livestock and humans. While Toxoplasma has long been recognized to cause neurologic disease in many warm blooded hosts, including humans, parasites in the related genus Sarcocystis also cause serious disease in livestock and even in humans. Some species of Sarcoystis closely resemble Toxoplasma and were until misdiagnosed as Toxoplasma. Although Toxoplasma will grow in many cell lines, Sarcocystis species are more difficult to propagate in culture. In the present paper, the authors provide step by step methods, including reagents and materials, required to grow Sarcocystis parasites in cell culture. The results will be of interest to biologists, and parasitologists.

Technical Abstract: More than 200 valid Sarcocystis species have been described in the parasitological literature. The developmental life cycle in the intermediate host and definitive host has only been described for a few species. The majority of species have been identified based solely on the presence of the sarcocyst stage in the muscles of the intermediate host. Information on the immune response to infection is limited due to difficulties growing and conducting experimental infections in laboratory animals. One genome has been assembled and only recently have molecular methods been used to examine phylogenetic relationships within, and between, different Sarcocystis species. Sarcocystis parasites are common pathogens infecting a wide range of animals, including humans, and this unit reviews the methods used for isolating infective stages of the parasite from both definitive and intermediate host(s), as well as methods used to initiate cultures from sporocysts and merozoites, and cryopreservation of various Sarcocystis spp. These methods are based on published reports and our experience with Sarcocystis species in cell culture over many years. The information presented is suitable for the efficient culture of many Sarcocystis species, however, some minor modifications may be needed based on the unique developmental patterns of some species.