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Protein profiles as an aid to taxonomy in the genus Diphyllobothrium.

Specific and distinct protein profiles were demonstrated by isoelectric focusing performed on soluble whole-body proteins of four Diphyllobothrium species. The protein patterns differed in number, position and relative density of the fractions separated. Altogether 31-36 protein bands per species were separated by isoelectric focusing in thin-layer polyacrylamide gels. The results show that sensitive chemotaxonomic methods can be used to help characterize and delimit species in the troublesome Diphyllobothrium group.

Animals

Comparison of restriction fragment length polymorphisms of ribosomal DNA between Diphyllobothrium nihonkaiense and D. latum.

Restriction fragment length polymorphisms (RFLPs) of ribosomal DNA (rDNA) were compared between Diphyllobothrium latum and D. nihonkaiense using seven kinds of restriction endonucleases. No intra-specific variation in restriction fragment profiles was shown within both species of Diphyllobothrium. Digestion of the genomic DNA with three endonucleases. SmaI, HinfI and HhaI, provided one or two different bands between two species, although the hybridization patterns generated with the others. HindIII, XbaI, StyI and HaeIII, were the same in both. RFLPs in the digested profiles with SmaI, HinfI and HhaI could be used as species-specific markers even if only fragments of strobilae with morphological similarity were available. Other cestodes, Spirometra erinacei and Taenia saginata, used as controls showed quite different restriction fragment patterns with all the enzymes used.

Animals

The development of the tapeworm Diphyllobothrium latum (L. 1756) (Cestoda; Pseudophyllidea) in its definitive hosts, with special references to the growth patterns of D. dendriticum (Nitzsch, 1824) and D. ditremum (Creplin, 1827).

When Diphyllobothrium latum develops from larva to adult in a definitive host, it first sheds the entire larval 'body' before growth of an adult strobila starts. This process of shedding off the entire larval abothrial extremity, piece by piece, takes about 48 h. By this time the larva has usually reached the anterior third of the small intestine of the host. D. dendriticum and D. ditremum develop quite differently, although exhibiting similar anterior migrations. In these two species the larvae develop directly into adults without the larval 'body' first being shed. The implications of the observed differences in growth pattern between these three species of Diphyllobothrium to the classification of diphyllobothriid cestodes is discussed briefly.

Aging

A marine Diphyllobothrium plerocercoid (cestoda, pseudophyllidea) from blue whiting (micromestius poutasson).

A type of Diphyllobothrium plerocercoid found in blue whiting, Micromestius poutasson, caught near the north edge of the Faeroe Banks, is described. The most dominant characters of these larvae are their typical dorsoventrally flattened body and their fully extruded and "adult"-like scolex. A morphological comparison with perocercoids of D. dendriticum, D. ditremum and D. latum is given.

Animals

Observations on the histogenesis of nervous tissue in Diphyllobothrium dendriticum Nitzsch, 1824 (Cestoda, Pseudophyllidea).

The mode of growth of the populations of cells within and immediately surrounding the main lateral nerve cords in the actively growing, immature Diphyllobothrium dendriticum was studied by the use of 3H-thymidine autoradiography. The population of nerve cells within the nerve cord grows only on account of cell migration from the surrounding parenchyma. No mitotic figures were observed in the nerve cords. The rate of growth is high. Within a period of cultivation for 2 days in hamster 34% of the nerve cells within the nerve cords have arrived from the parenchyma. These cells can be considered as cells at the starting point for differentiation into nerve cells. The protective layer of binding cells around the nerve cords also grows on a-count of cells migrating from the parenchyma. The binding cells actively move from the outer regions of the layer inwards close to t-e nerve cord. As stem cells for these types of cell differentiation serve the highly basophilic, actively dividing germinative cells.

Animals

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.

Animals

Tests with a new compound (Praziquantel) against Diphyllobothrium latum.

Praziquantel (Droncit) is a new anthelminthic agent with a strikingly wide therapeutic index. The efficiency of the drug against Diphyllobothrium latum was tested in vivo and in vitro. With a drug dose of 50 mg/kg, artificially infected hamsters were completely cured. The threshold value of the active dose was between 2-5 and 10-0 mg/kg. Clinical trials with Praziquantel against human diphyllobothriasis show great promise, all of ten patients treated with 25-0 mg/kg of the drug being cured. Praziquantel had little effect on larval or adult diphyllobothriids in vitro.

Animals

A hidden break in the 28.0S rRNA from Diphyllobothrium dendriticum.

Nondenatured and denatured total RNA from the tapeworm Diphyllobothrium dendriticum (Cestoda) was analysed by agarose gel electrophoresis. It was found that the large subunit ribosomal RNA (lrRNA) is 28.0S and the small subunit ribosomal RNA (srRNA) is 19.5S. Following denaturation the 28.0S rRNA was disrupted into a 19.5S subfragment and a 20.7S subfragment due to the presence of a centrally located hidden break. By hybridization of Northern blot membranes with oligonucleotide probes specific for the 5'- and 3'-ends of the lrRNA respectively, we have shown that the 19.5S subfragment is from the 5'-end (the alpha-subfragment) and the 20.7S subfragment from the 3'-end (the beta-subfragment) of the 28.0S rRNA of D. dendriticum.

Animals

Egg size and form as taxonomic criteria in Diphyllobothrium (Cestoda, Pseudophyllidea).

The size and form (length, width, and length: width ratio) of eggs of Diphyllobothrium dendriticum, D. ditremum and D. latum vary considerably among individual worms within each species. The size of eggs varies with host species and a decrease in egg size with increasing intensity of infestation is indicated. The egg size of D. latum increases during the first 10-12 days of egg production. For single worm infections in golden hamsters the mean egg length and width of D. ditremum are significantly smaller than the corresponding means of D. dendriticum and D. latum, while D. latum has significantly wider eggs than D. dendriticum. As taxonomic characteristics, egg size and form may contribute to species delimitation at the population level. For identification at the individual level the best possible accuracy is about 80%. This accuracy is considerably reduced when variation in host species and intensities of infestations are introduced. Scanning electron microscope studies did not reveal any differences among eggs of the three species.

Animals

Sequential development of the immune response in rainbow trout [Oncorhynchus mykiss (Walbaum, 1792)] to experimental plerocercoid infections of Diphyllobothrium dendriticum (Nitzsch, 1824).

Development of the inflammatory response of rainbow trout to experimental infections with Diphyllobothrium dendriticum plerocercoids is described using light and electron microscopy. The cellular response to plerocercoids occurred within 2 weeks post-infection (p.i.). This was followed by an increase in leucocyte numbers during weeks 3-6 p.i., with full encapsulation of plerocercoids by week 6 p.i. Neutrophils were the first leucocytes to engage the developing plerocercoid, followed by large influxes of macrophages which transformed into epithelioid cells. With longer times p.i. the accumulation of different leucocyte types increased, and a blood vascular network developed. Full development of the composite cyst was characterized by fibroplasia, particularly at the periphery of the cyst, and the subsequent deposition of a collagenous tissue matrix. Enzyme-linked immunosorbent assay (ELISA) examination of serum samples taken over the 20 week period showed that specific anti-D. dendriticum antibody titres were first detected at 5 weeks p.i. and increased to a maximum by 11 weeks p.i.

Animals

The laboratory maintenance of Diphyllobothrium dendriticum (Nitzsch, 1824).

A protocol for the routine laboratory maintenance of the life-cycle of Diphyllobothrium dendriticum, using herring gulls (Larus argentatus), Cyclops abyssorum and rainbow trout Oncorhynchus mykiss ( = Salmo gairdneri) as the definitive, first and second intermediate hosts respectively, is described.

Animals

Seasonal changes in the prevalence of ova of Diphyllobothrium ursi and Baylisascaris transfuga in the feces of the black bear (Ursus americanus).

An analysis of 114 samples of feces from black bears (Ursus americanus) from La Verendrye Park, Quebec revealed that the prevalence of eggs of Diphyllobothrium ursi and Baylisascaris transfuga varied seasonally. D. ursi ova were most prevalent during the summer and fall and at their lowest in spring while the converse was true for B. transfuga. This phenomenon possibly is associated with the cessation of feeding during the winter denning of black bears.

Animals

Ultrastructural studies on Diphyllobothrium ditremum and D. dendriticum (Cestoda, Pseudophyllidea), with emphasis on the scolex tegument and the tegument in the area around the genital atrium.

The scolices of D. ditremum and D. dendriticum are similar in regard to general morphology and ultrastructure of the tegument. At the scolex apex and along the upper bothrial edges sensory endings are numerous. The inner bothrium surface of D. ditremum appears to be plain or smooth, the distal cytoplasm is only about 0.6 mum thick and covered with long and slender microtriches. When D. ditremum is attached to the host gut an adhesive layer of secreted substance exists between the inner bothrium surface and the host intestinal villi. In D. dendriticum, however, no secreted layer exists between the inner bothrium surface and the host intestinal villi and the inner bothrium surface of this species appears to be lobed or lappet formed. The lobes are protrusions of the distal cytoplasm and are covered with long and slender microtriches. Large conglomerations of secretion globules appearing to be enclosed by a double membrane, are observed in the scolices of D. dendriticum and D. ditremum, although more frequent in the latter. Differences existing between the distal cytoplasm of the scolex and the gravid proglottid are described. The papillae around the genital atrium in mature and gravid segments were studied. Each papilla appears as a thickening of the distal cytoplasm, which in this region is 10-12 mum thick, when studied with transmission electron microscope. From the area between the papillae sensory endings appearing in sections to be either single, double or triple are described.

Animals