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Excystment and in vitro cultivation of Echinoparyphium serratum

Published online by Cambridge University Press:  06 April 2009

M. J. Howell
M. J. Howell
Affiliation:
Department of Zoology, The Australian National University, Canberra, Australia
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Extract

Seventy-five per cent of Echinoparyphium serratum metacercariae excysted within 10 min in a trypsin/sodium cholate solution after successive pretreatments with pepsin and sodium dithionite. Excystment appears to be an active process as the cyst wall is unaffected by the treatment except in the vicinity of the ‘escape aperture’.

Optimum development to sexual maturity in vivo required 72 h and was divided into six easily recognized stages against which development in vitro could be judged. These were undifferentiated (excysted metacercariae), cell multiplication, organogeny, gametogeny, vitellogenesis and oviposition. Migration of the juveniles from the lower regions of the intestine to the duodenum in the first few hours after infection was noted.

Cultures were prepared from excysted metacercariae and a variety of media were used. The beneficial effects of shaking cultures were apparent. No development took place in media containing saline, Parker 199 medium, homologous and heterologous sera, chicken embryo extract, albumen and glucose, either alone or in various combinations, although, relative to saline or Parker 199 medium controls, survival times were increased in some and decreased in others. Survival in some media above pH 7 was longer than below pH 7. Serum substrates in culture vessels, and the presence of additional glucose, did not increase longevity.

Development to organogeny was achieved using a yolk/albumen/saline or 199 medium, and addition of yeast extract to this favoured the later stages of development up to and including vitellogenesis. In general only a small percentage of juveniles in each culture reached vitellogenesis, and the amount of egg-shell protein formed and the size of the worms was always less than in vivo. Furthermore, development in vitro was much more prolonged than in vivo. Variability in development in vitro may be due to failure of the excystment procedure to satisfy completely the physiological ‘triggers’ which are probably necessary for normal development.

In some specimens vitellogenesis was reached without production of sperm. In others, production of eggs was thought to be inhibited by either a general deterioration of the juveniles or premature ‘tanning’ of the egg-shell protein, as significant amounts of egg-shell protein, sperm, and apparently normal ova were present.

The results of culturing E. serratum are compared briefly with those of D. phoxini.

I should like to record my sincere thanks to Professor J. D. Smyth for his advice and comments on the manuscript.

Type
Research Article
Information
Parasitology , Volume 58 , Issue 3 , August 1968 , pp. 583 - 597
Copyright
Copyright © Cambridge University Press 1968

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