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Serologic Response in a Marine Worm, Sipunculus Nudus¹

From the Department of Pathobiology, Johns Hopkins School of Hygiene and Public Health, and Station Biologique, Roscoff, France

Abstract

1. Earlier experiments had shown that injection of crab blood containing large numbers of a ciliate parasite, Anophrys magii, into a marine worm, Sipunculus nudus, was followed by the development of a lysin for the ciliate in the plasma.

2. Lysis of the ciliate was first apparent in a slower rate of swimming, then erratic motion, cessation of forward motion, all within a minute. The ciliate then fell to the bottom of the fluid, the shape changed to a rounded balloon form with a swollen posterior vacuole, the cilia disappeared and the whole ciliate superficially resembled a cyst. Lysis continued and in strongly positive plasmas, the entire ciliate disintegrated within 15 min.

3. The lysin was destroyed by heating to 45°C for 5 min, and was inactivated by ether. It was "used up" by repeated additions of Anophrys to the plasma. The presence of bacteria did not inhibit the test.

4. The response, i.e., the development of the lysin, lasted 5 to 7 days in most cases, and was evoked repeatedly, but did not persist.

5. A regular but briefer response was evoked by injection of fresh broth cultures of Vibrio sp. or large amounts of heat-killed bacteria.

6. No response was evoked by injecting fresh or sterilized sea water or a preparation of endotoxin.

7. The release of an apparently identical substance from the blood cells of Sipunculus was evoked in vivo within 15 min of injection of either Anophrys or bacteria. This released substance apparently persisted in the plasma only for a few hours after the injection, and then 1 or 2 days later the lysin reappeared and persisted for 5 to 7 days.

8. The release of lysin was also evoked by in vitro addition of Anophrys to freshly drawn blood.

9. The in vivo release of lysin from cells was regularly accompanied by the failure of blood to form irregular loose clots of several types of blood cells, by a number of changes in the cells, including a breakdown of innominate cells and a release of crystalline substance from these cells. However, in animals which had lysin again for several days after the injection, innominate cells with crystals were frequently found.

10. A spontaneous disease, apparently caused by a flagellate which burrowed into surface ulcerations and papules, was accompanied by a high titer of lysin.

11. All Sipunculus nudus were negative for lysin if tested within 1 to 2 days of capture, but about a third became positive 1 to 2 weeks after capture. By 14 days, laboratory-"adapted" (recovered?) worms were negative.

12. Repeated bleedings of such negative laboratory-adapted animals did not provoke the appearance of the lysin.

Footnotes

¹ This work was carried out during the tenure of a Guggenheim Fellowship.

I am indebted to Professor G. Teissier for the facilities of the laboratory at Roscoff.

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