1. Eggs of the potato cyst-nematode (Heterodera rostochiensis Woll.) were isolated by sieving a suspension of crushed cysts. Eggs were broken open by ultrasonic vibration and the egg shells separated from the released larvae by centrifuging in a potassium tartrate density gradient. About 1 mg. of dried egg shells was obtained from 1000 cysts. 2. The major constituent of the egg shells was protein (59%, calculated from nitrogen content). About 80% of the egg shells went into solution on acid hydrolysis. Of the 18 amino acids determined with the Technicon Auto-Analyser, proline was most abundant and, with aspartic acid, glycine and serine, made up about 64% by weight of the total amino acids. The small amounts of aromatic and sulphur-containing amino acids, and the presence of hydroxy-proline, indicate a collagen-like protein. 3. The egg shells gave a positive van Wisselingh colour test for chitin, and glucosamine was detected in their acid hydrolysate by chromatography. The glucosamine content of the egg shells, determined by the Elson-Morgan colorimetric method, was 7%, corresponding to about 9% chitin. 4. Dried egg shells contained about 7% of lipid, 6% of carbohydrate and 3% of ash. Polyphenols (3% by weight of the egg shells) were detected in the acid hydrolysates. 5. Neither the collagen nor the chitin showed evidence of crystallinity when examined by X-ray diffraction.
Soil containing new-generation cysts of Heterodera rostochiensis was taken from the field at monthly intervals during late summer and autumn and kept in various conditions for up to a year. The number of eggs that hatched in the stored cysts was compared each month with the number that hatched in cysts taken directly from the field. Eggs did not hatch readily when stimulated during the late autumn and early winter, although more did so in cysts taken from the field before August than after. A few more eggs hatched in cysts stored in air-dried soil than in cysts stored in moist soil. Some cysts were kept at 15 or 20 "C continuously and others at 5 , 15 or 30 "C for 6 weeks followed by 20 "C continuously. Storage at 30 "C caused eggs to hatch sooner, but otherwise the temperature of storage had little effect on hatch at any time of the year. Warmth also increased the hatch of H . cruciferae sooner, and some synthetic hatching agents did so with both of these species. When freed from new cysts, more eggs of H. rostochiensis hatched than in intact cysts and hatch was further increased when the fragments of tanned cystwall were left with the freed eggs. Puncturing the cyst-wall of new brown cysts had little effect on the hatch in potato root diffusate. Like eggs in new cysts, those in I-year-old cysts stored out of doors ceased to hatch during the autumn and winter. The term 'dormancy' is inadequate to describe the inability of eggs of H. rostochiensis and other Heterodera spp. to hatch in the appropriate stimulant and the term 'facultative diapause', as applied to insects, better fits the phenomenon.
An automated, quantitative microhemagglutination assay for antibodies to Treponema pallidum was developed by using T. pallidum-sensitized erythrocytes and an automatic serial-dilution instrument. Reactivity was found in sera from 54 rabbits and 6 chimpanzees infected with T. pallidum. Reactivity was also found in sera from animals infected with T. pertenue, T. carateum, and T. cuniculi. No reactivity was found in sera from 75 normal rabbits or from 129 rabbits immunized with cultivatable treponemes or a variety of other bacteria. In approximately 3 min, 13 twofold serial dilutions of each of 8 preabsorbed sera and the addition of sensitized erythrocytes to each dilution were accomplished automatically. The automated assay can serve as a research tool in quantitating antibodies to pathogenic treponemes, and evaluation of its clinical usefulness seems warranted.
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