Under in vitro conditions, muscle larvae of Trichinella spiralis secreted minute amounts of a cysteine proteinase into the outer environment from the stichosome. The proteinase hydrolyzed azocoll at pH 5.0 but not a number of synthetic N-blocked and N-unsubstituted proteinase substrates at this pH. The reducing compound dithioerythritol enhanced the enzyme activity, but the thiol-blocking reagent sodium-p-hydroxymercuribenzoate (0.1 mM) was without effect. Phenylmethylsulfonyl fluoride (PMSF) (2 mM) and leupeptin (100 mM) produced partial and complete inhibition, respectively, whereas soybean trypsin inhibitor, pepstatin A, and 1,10-phenanthroline were non-inhibitory. Calcium (1 mM) produced a slight decrease in the activity that was reversed by 1 mM EGTA. Although multiple proteinase activities were detected histochemically in the somatic muscles, stichosome, midgut, and genital primordium of the muscle larvae, none of these enzymes appeared to be the one secreted. Several histochemically demonstrable proteinases were also found in the cells of 48- to 72-h-old juveniles of the parasite. One was localized in the esophageal lumen and at or around the anterior esophagus of the larvae, where developing stichocytes are believed to occur. The proteinase hydrolyzed N-acetyl-L-methionine-L-naphthyl ester and was sensitive to the metal cation-complexing compound EGTA as well as to PMSF, an inhibitor of serine proteinases.
The nervous system of adult Diplostomum pseudospathaceum Niewiadomska, 1984 was studied using Koelle's (1951) method for revealing cholinesterase activity. The nervous system in the fore body grows but its pattern remains the same as in the metacercaria: three pairs of stems connected by numerous commissures and differentiated innervation of various organs in this part of the body. In the elongated hind body the nervous system develops according to another pattern: two pairs of stems connected by a number of commissures form a loose net surrounding the whole segment. The net is more dense at the body end around the genital opening and copulatory organs. A brief discussion of the development of the nervous system of D. pseudospathaceum from cercaria to adult stage is given.
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