Temperature effects on embryonic development and the life cycle of Diphyllobothrium dendriticum

Diphyllobothrium dendriticum eggs collected from hamster faeces were incubated at 10 or 20 degrees C, or maintained at 4 degrees C for 11-30 months. On day 65, 20-50% of eggs failed to hatch at 10 degrees C and 42-51% did not hatch by day 21 at 20 degrees C. Our study indicates that eggs begin to hatch in mid-August and persist until October in many lakes within the natural range of D. dendriticum. Our results demonstrate that eggs stored at 4 degrees C will hatch, suggesting they can persist in the environment for long periods and contribute to the D. dendriticum life cycle in high Arctic lakes by hatching months or years after release.     

Cytochemical characteristics of the labrocyte-like cells in the capsule of the plerocercoids of Triaenophorus nodulosus (Pallas, 1781) and Diphyllobothrium dendriticum (Nitzsch, 1824) cestodes]

Out of six fish species examined (Esox lucius, Paracottus kessleri, Perca fluviatilis, Leuciscus leuciscus baicalensis, Coregonus autumnalis migratorius, Thymallus arcticus baicalensis) labrocyte-like cells were detected in Esox lucius, Leuciscus leuciscus baicalensis, Coregonus autumnalis migratotius. The cells contain large metachromatic granules. Histochemical methods revealed in them sulfated and carboxylic acid mucopolysacharides. The amount of labrocyte-like cells in the capsules around plerocercoids of Triaenophrus nodulosis from Esox lucits liver depends on the age of the capsule and that of the plerocercoid. It increases when the connective tissue papsule is forming and decreases again under aging and degeneration of the capsule containing plerocercoid. The amount of labrocyte-like cells in the cestodes capsules of Raillietina increases sharply when they locate in an accidental organ (liver). This demonstrates the acuteness of organ reactivity to a nonspecific helminth.     

Immunocytochemical demonstration of neuropeptides and serotonin in the tapeworm Diphyllobothrium dendriticum

Summary The present immunocytochemical study concerns the distribution of four neuropeptides, FMRF-amide, vasotocin, leu-enkephalin and neurotensin, and of the bioamine serotonin in the plerocercoid larva of Diphyllobothrium dendriticum. Anti-FMRF-amide and vasotocin-reactivity occurs in perikarya and nerve fibres in the CNS and PNS of this worm. The peptide-containing fibres surround and seem to innervate the musculature and to terminate beneath the basal lamina of the tegument at the inner surface of the bothridia, suggesting a neurotransmitter function. Antileu-enkephalin reaction occurs in perikarya and fibres in the main nerve cords and in the PNS. Anti-neurotensin reactive fibres were observed in the neuropile of the nerve cords. Serotonin immunoreactivity was found in neurons in the ganglionic commissure of the brain and along the main nerve cords. This study is the first immunocytochemical identification of neuropeptides and serotonin in a parasitic flatworm and the information gained may be of importance for the development of new antihelminthics.     

The Nervous System of Early Larval Stages of the Cestode Diphyllobothrium dendriticum

The nervous system (NS) of the coracidium and procercoid larval stages of the pseudophyllidean cestode Diphyllobothrium dendriticum was examined by whole mount immunocytochemistry and electron microscopy. Antisera against the bioamine serotonin and the molluscan cardioexcitatory peptide FMRF-amide gave a positive immunoreaction in the NS of the procercoid. Ultrastructurally, three types of nervous elements were discerned. Ultrastructural evidence of both aminergic and peptidergic neurons in the coracidium is provided. The NS of these early larval stages is simply organized. The serotonergic part of the NS shows a beginning centralization in the procercoid, while the peptidergic neurons are more peripherally distributed.     

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.     

On the existence of non-cycling germinative cells in human epidermis in vivo and cell cycle aspects of psoriasis

Abstract. This report deals with the controversies of whether all germinative epidermal cells in human epidermis are in the cycling state and whether stimulated hyperproliferation of psoriatic epidermis is due to a shortening of the cell cycle time or to a recruitment of non-cycling germinative epidermal cells. Experiments were performed on human subjects in vivo . Continuous infusion of [³H]thymidine for 8½ days indicated that 40% of germinative epidermal cells reside in the non-cycling state. Proliferative stimulation by tape stripping indicated recruitment of non-cycling (G₀) germinative epidermal cells in both normal and psoriatic skin, and a prolongation (rather than a shortening) of cell cycle traverse in activated psoriatic epidermal cells.     

The distribution of F-actin during the development of Diphyllobothrium dendriticum (Cestoda)

The distribution of actin filaments in all developmental stages of the tapeworm Diphyllobothrium dendriticum was studied. It is the first investigation of the placement of microfilaments during the development of a flatworm, and the results show that actin filaments, in all developmental stages, can be found in the subtegument and the flame cells. Muscle fibers possibly corresponding to the longitudinal, transversal, and dorsoventral muscles of the adult and plerocercoid were already detected in the procercoid. Concerning the adult worm, a new set of longitudinal fibers in the peripheral parts of the adult proglottid was found. The ducts of the protonephridial system and the vitellarias were seen to be surrounded by longitudinally oriented actin filaments, while the uterine ducts and the vagina were encircled by microfilaments. Prominent layers of circular muscle fibers surrounded the cirrus and the seminal vesicle, and radial fibers were also detected. Areas faintly stained with TRITC-phalloidin were found in the developing germ cells, the cells of the genital anlage, vitelline cells, the tegument, and the main nerve cords. None of these structures were autofluorescent, which is also true concerning the intensively labeled oncospheral hooks.     

Observations on the Extracellular Spaces and Intercellular Junctions in Diphyllobothrium dendriticum (Cestoda)

Extracellular spaces and intercellular junctions were studied in adult Diphyllobothrium dendriticum after lanthanum nitrate infiltration. The parasitic tapeworms absorb nutrients across the whole body surface, the syncytial tegument. The results indicate that the distribution of nutrients can occur both in the extracellular stroma and in the intracellular compartment, through intercellular junctions of the gap junction-type. The fibrous basal lamina of the tegument forms a large extracellular compartment communicating through narrow interstices in the subtegument with extracellular lacunae in the parenchyma. Intercellular junctions of the gap junction-type frequently occur between the tegument cell bodies, as well as between tegument and parenchymal cells. According to the function of these cells in nutrient uptake and storage, metabolic coupling seemed the most likely mission for these junctions. Thus it can be concluded that the extracellular spaces and the intercellular junctions create the possibilities for an efficient distribution of nutrients in this tapeworm.     

Sequence analysis of an actin isoform in the flatworm Diphyllobothrium dendriticum (Cestoda)

We have examined actin cDNA of the flatworm Diphyllobothrium dendriticum (Cestoda). Actin is a contractile protein that has been implicated in a variety of developmental and cellular processes. It is highly conserved and present in all eukaryotic cells. It is of particular interest to analyze evolutionary preserved genes in flatworms, because ancestral flatworms are regarded to play a central role in the evolution of the metazoans (Barnes et al., 1998). Screening a cDNA library of D. dendriticum (UniZap XR, Stratagene) with a human -actin probe resulted in several positive clones. One of the cDNA inserts, Didactl, consisting of 1392 bp was completely sequenced. The established nucleotide sequence revealed a 5 untranslated region of 33 bp, the entire open reading frame of 1128 bp and a 3 untranslated region of 231 bp which ends in a stretch of 21 A residues. The potential polyadenylation signal (AATAAA) is located 14 bp upstream of the poly (A) tail. The deduced amino acid sequence of Didactl is 376 amino acids long. It is a typical invertebrate actin (Fyrberg et al., 1981) resembling more the cytoplasmic than the muscular isoforms of vertebrate actins. Didactl is for example 96% homologous to human cytoplasmic -actin but only 92.6% identical with human smooth muscle -actin. The actin proteins are generally encoded by a multigene family which differs in size from species to species. Most organisms have four to eight genes coding for actin in their genome, but the number of actin genes can also be over 20 (Hamelin et al., 1988). Sequence comparisons of Didactl and the partly sequenced cDNA clones indicate that D. dendriticum has at least four different genes coding for actin in its genome.     

Fatty acid makeup of the cestodes, Eubothrium crassum and Diphyllobothrium dendriticum]

The fat-acidic composition of E. crassum and D. dendriticum was investigated. Lipids of E. crassum differ in greater unsaturation as compared to these of D. dendriticum as well as in greater amount of acids of type omega 3, whereas acids of type omega 6 and stearic acid were found in extracts of E. crassum in less quantity than in D. dendriticum. This phenomenon is characteristic of organisms living under conditions of low temperatures.     

Genetic and morphological variability in a population of Diphyllobothrium dendriticum (Nitzsch, 1824)

Plerocercoids of Diphyllobothrium dendriticum were isolated from whitefish (Coregonus clupeaformis) from McGregor Lake, Alberta. Adult D. dendriticum were obtained by infection of golden hamsters (Mesocricetus auratus). Analysis of morphological variables indicated high variability and some overlap with D. latum. Correlations in morphology were due to the covariance of shape variables. Isozyme analysis also showed high variability, with two thirds of the enzymes studied being polymorphic. Correlations with morphology indicate a relationship between rare patterns for malate dehydrogenase and esterase and increased size of adults. Southern hybridisation with ribosomal gene probes produced two common patterns and a rare intermediate form, indicating that the species is variable even within a local population. Correlations between rDNA pattern type and morphological variables supported the hypothesis of a genetic base for the morphological variability.     

Cestode egg production dynamics and fertility in Diphyllobothrium dendriticum (Cestoda, Pseudophyllidea)

Dynamics of egg production of the tapeworm Diphyllobothrium dendriticum has been estimated experimentally in nestlings of the herring gull Larus argentatus per day and per reproductive period. The number of eggs in strobiles has been estimated for the maturation period. Mean egg production of the tapeworm per day (10.43 +/- 2.72 mln) is two orders of magnitude as high as the maximum number of eggs in a mature strobile (0.206 +/- 0.007). It is proposed to estimate coefficient of reproduction intensity as the ratio of egg output per unit of time and maximum egg number in a mature strobile.