GABA in the nervous system of the cestodes Diphyllobothrium dendriticum (Diphyllobothriidea) and Caryophyllaeus laticeps (Caryophyllidea), with comparative analysis of muscle innervation.

The nervous system (NS) of the cestodes Diphyllobothrium dendriticum (Diphyllobothriidea) and Caryophyllaeus laticeps (Caryophyllidea) was investigated using immunocytochemistry. The GABA neurotransmitter was identified in the NS of both species; GABAergic neurons were detected in the main nerve cords (MC). GABA-like immunoreactive neurons were predominantly unipolar and exhibited more intensive immunoreactivity in the neurite than in the perikaryon. In C. laticeps , GABA-like immunoreactive somas are located in both the MCs and peripheral NS near the longitudinal muscles. The innervation of the body musculature was studied using a combination of antibodies against GABA, serotonin (5-HT), and FMRFamide and with complementary staining of F-actin. In both species, the location of GABAergic neurites is associated with all muscle layers including the subtegumental, longitudinal, transverse, and dorsoventral muscles. The cytomorphology of 5-HTergic motoneurons in the MCs of both species is described and differences in muscle innervation between D. dendriticum and C. laticeps are demonstrated. No evidence for co-localization of GABA with 5-HT or FMRFamide neurotransmitters at any particular neuron was found. Neuropiles in MCs and peripheral NS had separate sets of immunoreactive neurites. A functional role for GABA in muscle innervation is discussed.

[Role of helminth antigens in the abnormal mitosis of bone marrow cells in laboratory animals].

The intraabdominal administration of somatic extracts of the cestodes Hydatigera taeniaformis Batsch 1786, Lamarck, 1816 and Diphyllobothrium latum Linnaeus, 1758 and the nematodes Anisakis simplex larva Rudolphi 1809, Toxocara canis Railliet et Henry, 1912 in albino mice proved that these helminths had a karyopathic effect on the bone marrow cells of the animals. The antigenic composition of these extracts was investigated using the agar gel immunodiffusion test. The antigenic composition of the parasites was ascertained to affect their karyopathic properties. The amount of antigens and their foreignness caused a marked karyopathic effect on the bone marrow cells of laboratory animals during intraabdominal administration.

Chemical structures and immunolocalization of glycosphingolipids isolated from Diphyllobothrium hottai adult worms and plerocercoids.

Glycosphingolipids (GSLs) were purified from adults and plerocercoids of the tapeworm Diphyllobothrium hottai, and their chemical structures were determined. Total lipid fractions prepared from chloroform/methanol extracts of whole tissues were fractionated successively on ion-exchange chromatography, silicic acid column chromatography, and preparative TLC. The purified GSLs were characterized by methylation analysis, TLC-immunostaining, liquid secondary ion MS, MALDI-TOF MS, and 1H-NMR. Ten GSLs were isolated from adult worms and four from plerocercoids, comprising mono-, di-, tri-, tetra-, and pentasaccharides. The GSL Gal beta 1-4(Fuc alpha 1-3)Glc beta 1-3Gal beta 1-Cer was found in adult worms but not in plerocercoids, whereas Ga lbeta 1-4 (Fuc alpha 1-3)Glc beta 1-3(Gal beta 1-6)Gal beta 1-Cer was found in both adult worms and plerocercoids. We previously found a similar series of GSLs in plerocercoids of the cestode Spirometra erinaceieuropaei, and termed them 'spirometosides'[Kawakami, Y. et al. (1996) Eur J. Biochem. 239, 905-911]. The core structure of spirometosides, Gal beta 1-4Glc beta 1-3 Gal beta 1-Cer, may have taxonomic significance, being characteristic of pseudophyllidean tapeworms. In the present study, GSL compositions were significantly different between adults and plerocercoids, and growth-dependent changes in composition were documented. We found a novel dihexosylceramide, Glc beta 1-3Gal beta 1-Cer, which is a possible precursor for spirometosides. Immunohistochemical examination showed that spirometoside GSLs are highly enriched in the inner surface of bothria, the major point of contact between the adult worm and the host's intestine. Our findings indicate that spirometosides are involved in host-parasite interaction.

[The effect of host temperature on the fatty acid composition of the total and membrane lipids in the cestodes Eubothrium crassum and Diphyllobothrium dendriticum]. / Vliianie temperatury khoziaina na zhirnokislotnyi sostav obshchikh i membrannykh lipidov tsestod Eubothrium crassum i Diphyllobothrium dendriticum.

A comparative qualitative and quantitative analysis of the fatty acid composition of total and lysosome membrane lipids of two cestode species Eubothrium crassum (from cold-blooded host) and Diphyllobothrium dendriticum (from warm-blooded host) has been carried out. It is shown that quantitative ratio of the fatty acids in the total and membranous lipids of the helminth of cold-blooded host was very similar whereas for the helminth of warm-blooded host fatty acid composition of membranous lipids was considerably more saturated compared with the one of total lipids. It is assumed that the high level of unsaturation of total lipids from D. dendriticum is associated with preadaptation mechanism of fatty acid spectra in parasites with the development coupled with a consecutive rapid change of different thermal regime habitats.

High-performance liquid chromatographic analysis of monoamines in the cestode Diphyllobothrium dendriticum.

The presence of biogene monoamines in adult and larval Diphyllobothrium dendriticum (Cestoda) was investigated by high-performance liquid chromatography with electrochemical detection (HPLC-ED). The biogene amines serotonin (5-HT), dopamine (DA), noradrenaline (NA), and adrenaline (A) as well as many of their precursors and metabolites, comprising a total of 15 different substances, were analyzed. 5-HT, DOPA, DA, NA, and A were detected in the worm, with 5-HT, DOPA, and DA being the dominating amines. The DA metabolites DOPAC and 3-MT or the 5-HT precursor 5-hydroxytryptophan could not be detected, but two unidentified substances, believed to be catecholic, were present in the worm. A high concentration of DOPA was measured in the proglottids and especially in the eggs. This is the first report of A in a flatworm.

Immunocytochemical evidence for the presence of substance P-like peptide in Diphyllobothrium dendriticum.

Substance P immunoreactivity (SP-IR) was detected in the nervous system of the gull-tapeworm Diphyllobothrium dendriticum. The distribution of the SP-IR neurons in the plerocercoid differs from that of other peptidergic and aminergic neurons in the worm. As well as occurring in the ganglionic commissure and along the two main nerve cords, SP-IR neurons are located laterally to the main nerve cords but not dorsally or ventrally. The SP-IR neurons have projections extending to the surface. Bipolar SP-IR neurons with processes to the surface also occur in the tip of the scolex. A sensory function for the SP-IR neurons is suggested.

Localization and identification of catecholamines in the nervous system of Diphyllobothrium dendriticum (Cestoda).

In the central and peripheral nervous system of larval and adult Diphyllobothrium dendriticum, catecholamines were detected using the glyoxylic acid-induced fluorescence method. High-pressure liquid chromatography (HPLC) analysis revealed the presence of dopa (about 65 ng/g fresh weight) and dopamine (5 ng/g fresh weight).

Leucocyte migration in rainbow trout (Oncorhynchus mykiss [Walbaum]): optimization of migration conditions and responses to host and pathogen (Diphyllobothrium dendriticum [Nitzsch]) derived chemoattractants.

A rainbow trout leucocyte-derived chemoattractant(s) was prepared and tested as a stimulant of leucocyte migration. It was used to optimize an in vitro leucocyte migration assay using a 48-well micro chemotaxis chamber. This assay has subsequently been used to test the chemoattractant activity of antigen extracts from the tegument of Diphyllobothrium dendriticum plerocercoids and conditioned medium obtained after in vitro maintenance of live plerocercoids. Leucocytes were found to have an increased directional motility (chemotactic response) to the host-derived chemoattractant(s) but a random increased motility (chemokinetic response) following stimulation/contact with parasite-derived antigens.