Hostname: page-component-cd9895bd7-gbm5v Total loading time: 0 Render date: 2024-12-26T00:57:23.787Z Has data issue: false hasContentIssue false
  • English
  • Français

Studies on tapeworm physiology

X. Axenic cultivation of the hydatid organism, Echinococcus granulosus; establishment of a basic technique

Published online by Cambridge University Press:  06 April 2009

J. D. Smyth
J. D. Smyth*
Affiliation:
Department of Zoology, Australian National University, Canberra, Australia
*
*Grateful acknowledgement is made for substantial technical assistance from Mrs H. Taylor, B.Sc., and Mr A. B. Howkins.
Get access Rights & Permissions [Opens in a new window]

Extract

1. A technique is described for the preparation of axenic cultures of protoscolices of Echinococcus granulosus.

2. Protoscolices removed aseptically from hydatid cysts are treated with 0·02% crystalline pepsin at pH 2·0 for 10 min. at 38°C. in order to remove (a) daughter cyst germinal membranes, and (b) dead or degenerating protoscolices.

3. Protoscolices may be maintained for more than 100 days in a variety of media. These include media containing various proportions of some of the following: chick embryo extract, beef embryo extract, hydatid fluid, bovine amniotic fluid, Parker 199, bovine serum.

4. 10–20 ml. of culture media was used at 38°C., either in roller tubes or shaken in a water-bath. Media were renewed every 48 hr.

5. Protoscolices developed into cysts by two routes, either (a) by becoming vesicular, gradually growing in size, and finally becoming enclosed in a laminated envelope, or (b) by forming a ‘posterior bladder’ which became relatively enormous and ultimately absorbed the scolex region, the whole forming a cyst within an envelope as in (a).

6. In the majority of media, after about 31–48 days' culture, a thin envelope was secreted which developed later into a thick laminated membrane.

7. Cysts with laminated membranes ultimately underwent fatty degeneration and became cytologically abnormal.

8. In the most successful experiments, cells or clusters of cells formed within the cysts and these may have represented the anlagen of the protoscolices.

9. In no case was segmentation, or any other sign of strobilar development, obtained.

10. These results show that the laminated membrane in E. granulosus is initially of parasite origin and not of host origin as believed by some workers to be the case.

11. Since there is evidence from the literature that E. multilocularis does not form such an envelope in vitro, a clear developmental difference appears to exist between the two species.

12. It is suggested that this difference offers an explanation for the formation of unilocular or multilocular cysts. Since E. granulosus secretes a laminated envelope, its development is limited to a single or unilocular type of cyst. In the case of E. multilocularis no such limitation occurs, so that the invasive multilocular type of cyst could more easily develop.

Type
Research Article
Information
Parasitology , Volume 52 , Issue 3-4 , November 1962 , pp. 441 - 457
Copyright
Copyright © Cambridge University Press 1962

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.)

Footnotes

†

This is a convenient term, recently introduced by some authors, for the rather unusual larvae of E. granulosus for which the term ‘scolex’ is not altogether appropriate.

References

REFERENCES

Bell, E. J. & Smyth, J. D. (1958). Cytological and histochemical criteria for evaluating development of trematodes and pseudophyllidean cestodes in vivo and in vitro. Parasitology, 48, 131–48.CrossRefGoogle ScholarPubMed
Berntzen, A. K. (1961). The in vitro cultivation of tapeworms. I. Growth of Hymenolepis diminuta (Cestoda: Cyclophyllidea). J. Parasit. 47, 351–5.CrossRefGoogle Scholar
Cameron, T. W. M. (1960). The development and experimental pathology of Echinococcus multilocularis and alveolar hydatid. Seventh International Congress of Hydatidosis (Rome, 1960).Google Scholar
Čmelik, S. (1952). Ein antigenes Polysaccharid aus den Echinococcuscysten. Biochem. Z. 322, 456–62.Google Scholar
Čmelik, S. & Briski, B. (1953). Untersucchungen über Eiweissfraktionen von Taenia echinococcus. Biochem. Z. 324, 104–14.Google Scholar
Coutelen, F. (1927 a). Essai de culture in vitro de scolex et hydatides échinococciques. Ann. Parasit. hum. comp. 5, 1–19.CrossRefGoogle Scholar
Coutelen, F. (1927 b). Sur l'évolution vésiculaire in vitro des scolex échinococciques. Ann. Parasit. hum. comp. 5, 239–42.CrossRefGoogle Scholar
Dévé, F. (1926). Archives de Médecine Expérimentale et d'Anatomie Pathologique, Tome 27, no. 2, Mai 1916.Google Scholar
Dévé, F. (1916). Évolution vesiculaire du scolex échinococcique in vitro. La culture artificielle du hyste hydatique. C.R. Soc. Biol., Paris, 94, 440–1.Google Scholar
Dévé, F. (1928). Scoliciculture hydatique en sac de collodion et in vitro. C.R. Soc. Biol., Paris, 98, 1176–7.Google Scholar
Dougherty, E. C. (1959). Introduction to ‘Axenic culture of invertebrate Metazoa: a goal'. Ann. N.Y. Acad. Sci. 77, 27–54.CrossRefGoogle Scholar
Flössner, O. (1925). Neue Untersuchungen üher die Echinokokkusfluessigkeit. 2 Mitteilung. Z. Biol., 82, 297–301.Google Scholar
Mueller, J. F. (1959). The laboratory propagation of Spirometra mansonoides (Mueller, 1935) as an experimental tool. III. In vitro cultivation of the plerocercoid larva in a cellfree medium. J. Parasit. 45, 561–9.CrossRefGoogle Scholar
Rausch, R. L. & Jentoft, V. L. (1957). Studies on the helminth fauna of Alaska. XXXI. Observations on the propagation of the larval Echinococcus multilocularis Leuckart, 1863, in vitro. J. Parasit. 43, 1–8.CrossRefGoogle ScholarPubMed
Schwabe, C. W., Schinazi, L. A. & Kilejian, A. (1959). Host parasite relationships in echinococcosis. II. Age resistance to secondary echinococcosis in the white mouse. Amer. J. Trop. Med. Hyg. 8, 29–36.CrossRefGoogle ScholarPubMed
Smyth, J. D. (1959). Maturation of larval pseudophyllidean cestodes and strigeid trematodes under axenic conditions: the significance of nutritional levels in platyhelminth development. Ann. N.Y. Acad. Sci. 77, 102–25.CrossRefGoogle Scholar
Weinstein, P. P. & Jones, M. F. (1959). Development in vitro of some parasitic nematodes of vertebrates. Ann. N.Y. Acad. Sci. 77, 137–62.CrossRefGoogle Scholar
Yamashita, J., Ohbayashi, M. & Kitamura, Y. (1958). Studies on Echinococcosis. VII. On the development of Echinococcus multilocularis in the tapeworm stage. Jap. J. Vet. Res. 6, 89–91.Google Scholar
Yamashita, J., Ohbayashi, M. & Konno, S. (1956). Studies on Echinococcosis. III. On experimental infection in dogs, especially on the development of Echinococcus granulosus (Batsch, 1786). Jap. J. Vet. Res. 4, 113–122.Google Scholar