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Cytological and histochemical criteria for evaluating development of trematodes and pseudophyllidean cestodes in vivo and in vitro

Published online by Cambridge University Press:  06 April 2009

E. J. Bell
Affiliation:
Department of Experimental Biology, Trinity College, Dublin, Ireland
J. D. Smyth
Affiliation:
Department of Experimental Biology, Trinity College, Dublin, Ireland

Extract

1. Maturation of the plerocercoid of the pseudophyllidean cestode Diphyllobothrium sp. and the metacercaria of the strigeid trematode Diplostomum phoxini follow a similar pattern, which may be divided into the following phases: (1) cell multiplication; (2) segmentation or body shaping; (3) organogeny; (4) early gametogeny; (5) late gametogeny; (6) egg-shell formation and vitellogenesis; (7) oviposition.

2. Cytological or histochemical criteria have been developed by means of which each developmental phase may be recognized. Development in vitro may be considered comparable with that occurring in vivo if the same phases are completed with the same incubation time and without the appearance of cytological abnormalities.

3. The cell multiplication phase is characterized by high mitotic activity. This activity in vitro can be evaluated by incubating larvae after 24 hr. culture for a standard time (5 hr. Diphyllobothrium; 4 hr. Diplostomum) in 10−4 colchicine and comparing the number of mitoses seen in a defined area of an aceto-orcein squash with those found in tissue matured in vivo for the same period. Use of this criterion enables the growth-producing properties of a medium to be assessed after 24 hr. culture.

4. Phases 2–4 are characterized by the appearance of the tubular genitalia, the rudiments of which are readily recognized in aceto-orcein squashes or whole mount preparations.

5. The presence of spermatids and mature spermatozoa (phases 4–5) may be detected in aceto-orcein squashes or in fresh tissue teases. The quantity and activity of spermatozoa also serve as important criteria.

6. Recognition of phase 6, egg-shell formation, is carried out by means of diazo reagents which give orange or orange-red reactions with the phenolic egg-shell precursors in the vitellaria, as seen in whole mount preparations. Abnormal egg-shell development is reflected in the failure of vitellaria to give a normal diazo reaction.

7. The appearance of eggs which are macroscopically normal constitutes the final maturation phase. Early detection of eggs (especially abnormal ones) is facilitated by the use of the diazo reaction or the catechol test for polyphenol oxidase.

8. Embryonation and hatching of eggs to produce morphologically normal infective coracidia or miracidia serve as a final criteria.

9. Criteria for the maintenance of the adult condition are also considered.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1958

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References

Archer, D. M. & Hopkins, C. A. (1957). Studies on cestode metabolism. III. The growth pattern of Diphyllobothrium sp. (Cestode: Pseudophyllidea) in a definitive host. Exp. Parasitol. (in the Press).Google Scholar
Bell, E. J. & Hopkins, C. A. (1956). The development of Diplostomum phoxini (Strigeida, Trematoda). Ann. Trop. Med. Parasit. 50, 275–82.CrossRefGoogle ScholarPubMed
Bell, E. J. & Smyth, J. D. (1957). Axenic development of Diplostomum phoxini (Strigeida, Trematoda). Unpublished work.Google Scholar
Brown, C. H. (1952). Some structural proteins of Mytilus edulis. Quart. J. Micr. Sci. 93, 487503.Google Scholar
Brown, C. H. (1955). Egg-capsule protein of Selachians and trout. Quart. J. Micr. Sci. 96, 483–8.Google Scholar
Clegg, J. (1957). Studies on the maintenance of Fasciola hepatica in vitro. Ph.D. Thesis. Univ. of London.Google Scholar
Dawes, B. (1940). The Trematoda. Cambridge University Press.Google Scholar
Ferguson, M. S. (1940). Excystment and sterilization of metacercaria of the avian strigeid trematode, Posthodiplostomum minimum and their development into adult worms in sterile cultures. J. Parasit. 26, 359–72.CrossRefGoogle Scholar
Gonnert, R. (1955). Schistosomiasis–Studien. II. Über die Eibildung bei Schistosoma mansoni und das Schicksal der Eier im Wirtsorganismus. Z. Tropenmed. u Parasit. 6, 3352.Google Scholar
Johri, L. N. (1957). A morphological and histochemical study of egg-shell formation in a cyclophyllidean cestode. Parasitology, 47, 2129CrossRefGoogle Scholar
Johri, L. N. & Smyth, J. D. (1956). A histochemical approach to the study of helminth morphology. Parasitology, 46, 107116.CrossRefGoogle Scholar
Kouri, P. & Nauss, R. W. (1938). Formation of the egg-shell in Fasciola hepatica as demonstrated by histological methods. J. Parasit. 24, 291310.CrossRefGoogle Scholar
Kuhlow, F. (1953). Über die Entwicklung und Anatomie von Diphyllobothrium dendriticum Nitzsch 1824. Z. Parasitenk. 16, 135.CrossRefGoogle ScholarPubMed
Lillie, R. D. 1954. Argentaffin and Schiff reactions after periodic acid oxidation and aldehyde blocking reactions. J. Histochem. Cytochem. 2, 123–36.CrossRefGoogle ScholarPubMed
Lushbaugh, C. C. (1956). Morphological methods of determining doubling times. J. Histochem. Cytochem. 4, 499507.CrossRefGoogle ScholarPubMed
Markov, G. S. (1943). The dynamic of reserve nutritive substances in parasitic worms in artificial media. Zool. Zh. 22, 318.Google Scholar
Rausch, R. L. (1956). Studies on the helminth fauna of Alaska. XXVIII. The description and occurrence of Diphyllobothrium dalliae n.sp. (Cestoda). Trans. Amer. Micr. Soc. 75, 180–7.CrossRefGoogle Scholar
Rohrbacher, G. H. (1957). Observations on the survival in vitro of bacteria-free adult common liver flukes, Fasciola hepatica Linn., 1758. J. Parasit. 43, 918.CrossRefGoogle Scholar
Romanini, M. G. (1947). Contributo alla conoscenza istochimica dei vitellogeni di distoma hepaticum. Monit. zool. ital. 56, 36.Google Scholar
Smyth, J. D. (1946). Studies on tapeworm physiology. i. Cultivation of Schistocephalus solidus in vitro. J. Exp. Biol. 23, 4770.CrossRefGoogle ScholarPubMed
Smyth, J. D. (1947). Studies on tapeworm physiology. III. Aseptic cultivation of larval Diphyllobothriidae in vitro. J. Exp. Biol. 24, 374–86.CrossRefGoogle Scholar
Smyth, J. D. (1949). Studies on tapeworm physiology. IV. Further observations on the development of Ligula intestinalis in vitro. J. Exp. Biol. 26, 114.CrossRefGoogle Scholar
Smyth, J. D. (1950). Studies on tapeworm physiology. V. Further observations on the maturation of Schistocephalus solidus (Diphyllobothriidae) under sterile conditions in vitro. J. Parasit. 36, 371–83.CrossRefGoogle ScholarPubMed
Smyth, J. D. (1951). Egg-shell formation in trematodes and cestodes as demonstrated by the methyl or malachite green techniques. Nature, Lond., 168, 322–3.CrossRefGoogle ScholarPubMed
Smyth, J. D. (1952). Studies on tapeworm physiology. VI. The effect of temperature on the maturation of Schistocephalus solidus in vitro. J. Exp. Biol. 29, 304–9.CrossRefGoogle Scholar
Smyth, J. D. (1954 a). Studies on tapeworm physiology. VII. Fertilization of Schistocephalus solidus in vitro. Exp. Parasit. 3, 6471.CrossRefGoogle ScholarPubMed
Smyth, J. D. (1954 b). A technique for the histochemical demonstration of polyphenol oxidase and its application to egg-shell formation in helminths and byssus formation in Mytilus. Quart. J. Micr. Sci. 95, 139–52.Google Scholar
Smyth, J. D. (1956 a). Studies on tapeworm physiology. VIII. Occurrence of somatic mitosis in Diphyllobothrium spp. and its use as a criterion for assessing growth in vitro. Exp. Parasit. 5, 260–70.CrossRefGoogle ScholarPubMed
Smyth, J. D. (1956 b). Studies on tapeworm physiology. IX. A histochemical study of egg-shell formation in Schistocephalus solidus (Pseudophyllidea). Exp. Parasit. 5, 519–40.CrossRefGoogle Scholar
Smyth, J. D. (1957). In preparation.Google Scholar
Smyth, J. D. & Howie, D. I. D. (1957). Unpublished work.Google Scholar
Stephenson, W. (1947). Physiological and histochemical observations on the adult liver fluke, Fasciola hepatica. L. III. Egg-shell formation. Parasitology, 38, 128139.CrossRefGoogle Scholar
Tandler, C. J. (1955). The reaction of nuclei with ammonacal silver nitrate in the darkness. J. Histochem. Cytochem. 3, 196202.CrossRefGoogle Scholar
Vialli, M. (1950). Fenoli e tannazione chinonica delle proteine. Monit. zool. ital. 58, 8387.Google Scholar
Weinstein, P. P. & Jones, M. F. (1956). The in vitro cultivation of Nippostrongylus muris to the adult stage. J. Parasit. 42, 215–36.CrossRefGoogle Scholar