Studies on resistance of sheep to infestation with Haemonchus contortus and Trichostrongylus spp. and on the immunological reactions of sheep exposed to infestation. I. The preparation of antigens for the complement fixation test and the reactivity of the biochemical fractions of H. contortus

Abstract

All antigen was developed to detect circulating antibodies by means of the complement fixation test in sheep infested with Haemonchus contortus and Trichostrongylus spp. Extraction of worm material at 100°C. for 10 minutes was found to be the most satisfactory method for the preparation of antigens. Potent antigens were prepared from young adult H. contortus, from third-stage infective larvae, and from the eggs. Old mature adult H. contortus yielded antigens of low potency. No significant difference was found between the potency of antigens prepared from male and female adult H. contortus collected from the same sheep. Both adult Trichostrongylus spp. and third-stage infective larvae consistently yielded antigens of high potency. No significant difference was found in the results obtained with larval or adult H. contortus antigens, adjusted to the same potency and tested with natural H. contortus antisera. H. contortus and Trichostrongylus spp. absorbed the antiserum to each other. It was shown that a lipid was an essential constituent of the boiled antigen in the complement fixation reaction with natural antisera. Lipid-free antigens from H. contortus failed to react with natural antisera. The lipid was not antigenic when injected into rabbits. The carbohydrate fraction of H. contortus did not fix complement in the presence of natural antisera and was not antigenic when injected into rabbits. The lipid fraction of a variety of nematode parasites reacted with natural antisera to H. contortus infestation. Similar lipid fractions of two species of trematodes did not react with natural antisera to H. contortus infestation. Normal saline suspensions of the lipid-free material from a variety of helminths showed a greater degree of specificity when tested with artificially prepared antisera than did the lipid fractions.