The occurrence of estrone and estriol in Trichostrongylus colubriformis (Nematoda).

Mammalian sex steroids and cholesterol were isolated from the lipid extract of the zooparasitic nematode Trichostrongylus colubriformis. In addition to the previously identified sex steroids progesterone and testosterone, estrone and estriol were detected and isolated from the mixture. The steroids were analyzed by thin-layer, gas-liquid and high-performance liquid chromatography, and their structures confirmed by proton nuclear magnetic resonance and mass spectroscopy.

Trichostrongylus colubriformis: analysis of monoclonal antibody and lectin binding to the larval cuticle.

A monoclonal antibody (VRI 86-1) raised against third stage Trichostrongylus colubriformis preferentially bound to the excretory pore of living exsheathed larvae, with little or no binding to other sites on the parasite surface. A similar binding pattern was observed with fluorescein-labelled wheat germ agglutinin (WGA), although the anterior end of the parasite was also stained. To gain information at the molecular level regarding the parasite components at these sites, the residues recognized by WGA (i.e. N-acetylglucosamine and N-acetylneuraminic acid) were radiolabelled on the surface of living larvae. After homogenization and detergent extraction of the larvae, four dominant bands and a number of minor bands were revealed by SDS-PAGE and fluorography. None of these bands was specifically immunoprecipitated or recognized on a Western blot by VRI 86-1, suggesting that the epitope recognized by this antibody either resides on a different molecule or is destroyed or changed during the radiolabelling procedures. These results provide further evidence that the nematode cuticle is not uniform at the molecular level, and that the excretory pore contains molecules and antigens that may be unique to that site.

Two-dimensional electrophoretic comparison of the antigens and biosynthetically labelled proteins of Trichostrongylus colubriformis and Ostertagia circumcincta.

The proteins and antigens of different life stages of Trichostrongylus colubriformis were compared with those of Ostertagia circumcincta in an attempt to identify the subset of parasite molecules that is genus-specific and that may therefore be involved in the induction of genus-specific, host-protective immunity. Novel short-term culture techniques were instituted to label biosynthetically the proteins of the infective larval and adult stages of the parasites using 35S-methionine. High resolution, two-dimensional electrophoretic profiles of the labelled proteins indicated that the majority of proteins synthesized by adults were also present in the larval stages. Qualitative differences in the levels of these common proteins were observed, indicating differences in protein expression or turnover. There was extensive homology between larvae from the different species, with only eight major differences apparent in their profiles of biosynthetically-labelled proteins. Western blot analysis using immune sheep sera indicated that extensive homology also existed between the antigens of T. colubriformis and O. circumcincta larvae.

Biogenic amines and their metabolites in Trichostrongylus colubriformis, a nematode parasite of ruminants.

The following biogenic amines (BA) and BA metabolites were identified via HPLC in homogenates prepared from adults of Trichostrongylus colubriformis (Nematoda) recovered from the intestines of goats: N-acetyldopamine, DOPA, dopamine, epinephrine, epinine, 5-hydroxyindoleacetic acid, 4-hydroxy-3-methoxy-phenylglycol, 3-(p-hydroxyphenyl) proprionic acid, metanephrine, O-methyl-DOPA, 3-methoxytyramine, norepinephrine, normetanephrine, octopamine, p-hydroxymandelic acid, serotonin, synephrine, tyramine and vanillylmandelic acid. The mean concentrations of these compounds in groups of worms collected from different goats did not differ significantly with sex, but between the groups variance was high with probable components of both host and nematode origin.