Hostname: page-component-cd9895bd7-p9bg8 Total loading time: 0 Render date: 2024-12-26T01:13:15.948Z Has data issue: false hasContentIssue false
  • English
  • Français

Eimeria tenella sporozoites: the method of excystation affects the surface membrane proteins

Published online by Cambridge University Press:  06 April 2009

M. H. Wisher  and
M. E. Rose
M. H. Wisher
Affiliation:
Institute for Animal Disease Research, Houghton Laboratory, Houghton, Huntingdon, Cambs. PE 17 2DA
M. E. Rose
Affiliation:
Institute for Animal Disease Research, Houghton Laboratory, Houghton, Huntingdon, Cambs. PE 17 2DA
Get access Rights & Permissions [Opens in a new window]

Summary

Eimerian sporozoites can be recovered from intestinal washings after oral administration of oocysts to chickens but suspensions of sporozoites are usually prepared in the laboratory by incubation of sporocysts or fractured oocysts in vitro, at body temperatures, with relatively high concentrations of trypsin and bile salts. Since these agents affect membrane structure, the surface membrane of proteins of Eimeria tenella sporozoites excysted in vivo and in vitro have been compared. Surface radio-iodination followed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) showed that more 125I was incorporated into polypeptides on sporozoites excysted in vivo than on sporozoites excysted in vitro. The 125I-polypeptide profile of sporozoites excysted in vivo was more resistant to subsequent incubation with pure trypsin than that of sporozoites excysted in vitro, but incubation with bile salts resulted in the loss of some iodinated polypeptides from both preparations of iodinated sporozoites. Reaction with combinations of crude trypsin and bile salts led to the lysis of sporozoites. The method of excystation had no effect on the reaction of convalescent chicken serum with Western blots of sporozoites but the results of immunofluorescent staining carried out with mouse monoclonal antibodies indicated that the structure of the cell surface was altered and some antigenic determinants were lost from sporozoites excysted in vitro. In contrast, neither the infectivity of sporozoites determined in vivo, nor their invasion of cultured cells was changed by the method of excystation.

Type
Research Article
Information
Parasitology , Volume 95 , Issue 3 , December 1987 , pp. 479 - 489
Copyright
Copyright © Cambridge University Press 1987

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

Augustine, P. C. & Danforth, H. D. (1985). Effects of hybridoma antibodies on invasion of cultured cells by sporozoites of Eimeria. Avian Diseases 29, 1212–23.CrossRefGoogle ScholarPubMed
Bird, F. H. & Moreau, G. E. (1978). The effect of dietary protein levels in isocaloric diets on the composition of avian pancreatic juice. Poultry Science 57, 1622–8.CrossRefGoogle ScholarPubMed
Chapman, H. D. (1978). Studies on the excystation of different species of Eimeria in vitro. Zeitschrift für Parasitenkunde 56, 115–21.CrossRefGoogle ScholarPubMed
Danforth, H. D. (1983). Use of monoclonal antibodies directed against Eimeria tenella sporozoites to determine stage specificity and in vitro effect on parasite penetration and development. American Journal of Veterinary Research 44, 1722–7.Google ScholarPubMed
Davis, L. R. (1973). Techniques. In The Coccidia (ed. Hammond, D. M. and Long, P. L.), pp. 411–58. Baltimore: University Park Press.Google Scholar
Doran, D. J. (1970). From sporozoites to oocysts in cell culture. Proceedings of the Helminthological Society of Washington 37, 84–92.Google Scholar
Doran, D. J. & Farr, M. M. (1962). Excystation of the poultry coccidium, Eimeria acervulina. Journal of Protozoology 9, 154–61.CrossRefGoogle ScholarPubMed
Farr, M. M. & Doran, D. J. (1962). Comparative excystation of four species of poultry coccidia. Journal of Protozoology 9, 403–7.CrossRefGoogle ScholarPubMed
Freer, S. M. (1984). A permanent wet-mount for fluorescent microscopy of surface stained lymphoid cells. Journal of Immunological Methods 66, 187–8.CrossRefGoogle ScholarPubMed
Helenius, A. & Simons, K. (1975). Solubilisation of membranes by detergents. Biochimica et Biophysica Acta 415, 29–79.CrossRefGoogle ScholarPubMed
Hurwitz, S., Bar, A., Katz, M., Sklan, D. & Budowski, P. (1973). Absorption and secretion of fatty acids and bile acids in the intestine of the laying fowl. Journal of Nutrition 103, 543–7.CrossRefGoogle ScholarPubMed
Köhler, G. & Milstein, C. (1975). Continuous cultures of fused cells secreting antibody of predefined specificity. Nature, London 256, 495–7.CrossRefGoogle ScholarPubMed
Russell, D. G. & Sinden, R. E. (1981). The role of the cytoskeleton in the motility of coccidian sporozoites. Journal of Cell Science 50, 345–59.CrossRefGoogle ScholarPubMed
Schwert, G. W. & Takenaka, Y. (1955). A spectrophotometric determination of trypsin and chymotrypsin. Biochimica et Biophysica Acta 16, 570–5.CrossRefGoogle ScholarPubMed
Speer, C. A., Wong, R. B., Blixt, J. A. & Schenkel, R. H. (1985). Capping of immune complexes by sporozoites of Eimeria tenella. Journal of Parasitology 71, 33–42.CrossRefGoogle ScholarPubMed
Wisher, M. H. (1986). Identification of the sporozoite antigens of Eimeria tenella. Molecular and Biochemical Parasitology 21, 7–15.CrossRefGoogle ScholarPubMed
Wisher, M. H. & Rose, M. E. (1984). The large-scale preparation of purified sporozoites of Eimeria spp. by metrizamide density-gradient centrifugation. Parasitology 88, 515–19.CrossRefGoogle ScholarPubMed