Hostname: page-component-77c89778f8-n9wrp Total loading time: 0 Render date: 2024-07-22T03:05:31.324Z Has data issue: false hasContentIssue false

Separation of Isospora (Toxoplasma) gondii cysts and cystozoites from mouse brain tissue by continuous density - gradient centrifugation

Published online by Cambridge University Press:  06 April 2009

A. W. C. A. Cornelissen
Affiliation:
Department of Tropical Veterinary Medicine and Protozoology, State University Utrecht, Biltstraat 172, 3572 BP Utrecht, The Netherlands
J. P. Overdulve
Affiliation:
Department of Tropical Veterinary Medicine and Protozoology, State University Utrecht, Biltstraat 172, 3572 BP Utrecht, The Netherlands
J. M. Hoenderboom
Affiliation:
Department of Tropical Veterinary Medicine and Protozoology, State University Utrecht, Biltstraat 172, 3572 BP Utrecht, The Netherlands

Summary

A simple, quick and reproducible method consisting of density-gradient centrifugation of homogenized infected mouse brain tissue on Percoll is described for the isolation and purification of cysts of Isospora (Toxoplasma) gondii. A 100% recovery of cysts, with 74·2% in a single fraction with a specific gravity of 1·056, was obtained by overlaying homogenates of infected mouse brains on a pre-formed Percoll gradient and centrifugation at low g forces. With this procedure recovery was independent of the age of the cysts. Titration of purified cystozoites showed there to be no loss of infectivity.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1981

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Dubey, J. P., Voller, A. & Frenkel, J. K. (1970). The Toxoplasma gondii oocyst from cat faeces. Journal of Experimental Medicine 132, 636–62.CrossRefGoogle Scholar
Masihi, K. N. & Jíra, J.(1975). Studies on purification of Toxoplasma gondii zoites by sintered glass filters. Acta parasitologica Polonica 23, 653–8.Google Scholar
Masihi, K. N., Mach, O., Valkoun, A. & Jíra, J. (1976). Toxoplasma gondii: large scale purification by zonal density gradient centrifugation. Experimental Parasitology 39, 84–7.CrossRefGoogle ScholarPubMed
Motomura, I. & Jo, K. (1970). Estimation of the number of parasites contained within a cyst, and growth and distribution of cysts in the brains of mice infected with Toxoplasma gondii. Tropical Medicine (Japan) 12, 4150.Google Scholar
Nakabayashi, T. & Motomura, I. (1968). A method for separating cysts of Toxoplasma gondii from the infected mouse brains by multi-layer centrifugation with Gum Arabic solution. Tropical Medicine (Japan) 10, 7280.Google Scholar
Overdulve, J. P. (1978). Excretion of Toxoplasma gondii by non-immunized and immunized cats; its role in the epidemiology of Toxoplasmosis. Proceedings of the Koninklijke Nederlandse Akademie van Wetenschappen, Series C 81, 118.Google Scholar
Overdulve, J. P. & Antonisse, H. W. (1970). Measurement of the effect of low temperature on Protozoa by titration. 1. A mathematical model for titration, using prepatent period or survival time; with a discussion of the method of the ID63. Experimental Parasitology 27, 310–22.CrossRefGoogle ScholarPubMed
Pettersen, E. K. (1979). Destruction of Toxoplasma gondii by HCL solution. Acta pathologica microbiologica Scandinavica, Section B 87, 217–20.Google Scholar
Tryon, J. C., Weidner, E., Larson, A. D. & Hart, L. T. (1978). A rapid method for isolating purified Toxoplasma tachyzoites from peritoneal exudate and cell culture. Journal of Parasitology 64, 1127–9.CrossRefGoogle ScholarPubMed
Van der Waay, D. (1959). Ontwikkeling en transmissie-mogelijkheden van Toxoplasma gondii in experimentele infectie met chronisch beloop bij muizen. Thesis, Leiden, pp. 48.Google Scholar
Work, K. (1971). Toxoplasmosis. With special reference to transmission and life cycle of Toxoplasma gondii. Acta pathologica microbiologica Scandinavica, Section B, Suppl. 221, 1518.Google ScholarPubMed