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.     

The immune response of wild rainbow trout, Salmo gairdneri Richardson, to naturally acquired plerocercoid infections of Diphyllobothrium dendriticum (Nitzsch, 1824) and D. ditremum (Creplin, 1825)

Natural infections of rainbow trout with two species of Diphyllobothrium result in a host inflammatory response encapsulating the plerocercoid. The encapsulating cyst, observed by light and electron microscopy, comprises leucocytes, fibroblasts and collagenous connective tissue and is infiltrated with a blood vascular network. An indirect immunofluorescence technique and enzyme-linked immunosorbant assay (ELISA) have shown that specific antibodies are elicited by the fish host to these Diphyllobothrium spp. These antibodies have been semi-quantitatively measured by ELISA and correlated with worm burdens in individual fish.     

The abundance and distribution of Diphyllobothrium dendriticum (Nitzsch) and D. ditremum (Creplin) in the char Salvelinus alpinus (L.) in Sweden

A total of 654 char, Salvelinus alpinus (L.) and 33 brown trout, Salmo trutta L., were examined for the plerocercoids of Diphyllobothrium dendriticum (Nitzsch, 1824) and D. ditremum (Creplin, 1825). Only char were found to be infected. Both parasitic species were abundant. The incidence and mean intensity of infection for D. dendriticum were 83.2% and 8.8, respectively; for D. ditremum 93.7% and 45.0. There was no variation in the intensity of infection between different parts of the lake. The incidence and intensity of infection increased with the age of the fish to age 8+ for both species. From this age the intensity of infection leveled off or decreased. No differences in intensity of infection between the sexes were found. The frequency distribution of parasite counts were overdispersed and fitted the negative binomial distribution even within single age-groups. The two parasite species showed a high positive correlation in intensity of infection, D. ditremum was concentrated in the stomach and pyloric caeca. D. dendriticum was more widely distributed within the host and was supposed to kill heavily infected fish.     

The dynamics of infection of Diphyllobothrium dendriticum (Nitzsch) and D. ditremum (Creplin) in the char Salvelinus alpinus (L.) in Sweden

A total of 654 char, Salvelinus alpinus (L.), from Lake Bjellojaure, Sweden, were examined for plerocercoids of Diphyllobothrium dendriticum (Nitzsch, 1824) and D. ditremum (Creplin, 1825) from September 1969 to December 1974. Plankton samples showed that the dominating copepod species in the lake were Eudiaptomus graciloides (Lilljeborg), Cyclops scutifer Sars and Heterocope appendiculata Sars. E. graciloides passed the winter as adults and C. scutifer as copepodids while H. appendiculata was restricted to a short period in the summer. Copepods occurred in the stomachs in September-October and to a certain extent in April-May, but only cyclopids and Heterocope sp. were found. C. scutifer is proposed to be the main first intermediate host of both D. dendriticum and D. ditremum . The potential final hosts were thecommon gull, Larus canus L., for D. dendriticum and the black-throated diver, Gavia arctica (L.), for D. ditremum . One common gull was experimentally successfully infected with plerocercoids of D. dendriticum . The parasites showed no seasonal cycle in incidence of infection.     

Differentiation of the tegument and associated structures in Diphyllobothrium dendriticum Nitsch (1824) (Cestoda: Pseudophyllidea). An electron microscopical study

Grammeltvedt Anne-François 1973. Differentiation of the tegument and associated structures in Diphyllobothrium dendriticum Nitsch 1824 (Cestoda : Pseudophyllidea). An electron microscopical study. International Journal for Parasitology3: 321–327. The differentiation of the tegument and associated structures of the coracidium, procercoid, plerocercoid and adult is described. The embryophore is composed of four zones and is covered by a fibrous layer resembling a glycocalyx. The oncospheral plasma membrane is extensively folded. A typical cestode tegument, with a distal and perinuclear cytoplasm, is probably already existing in the coracidium. The formation of the microvilli starts after about three days in the copepod host. In young procercoids ribosomes and Golgi complexes were observed in the distal cytoplasm. These organelles disappear at later stages. The infective procercoid has a typical tegument. The microvilli are shaped like a thorn compressed from the sides. They have an electron dense tip and a less dense base in which microfilaments are seen. Bodies, called disc-shaped and lamellated bodies, are described. The microvilli of the plerocercoid are characterized by a great variation in shape. The villi are bounded by two unit membranes. The lamellated bodies are especially well developed. The adult microvilli are uniform in shape. The lamellated bodies are few in young adults and disappear in mature worms.     

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.     

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.     

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.     

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.     

Release of neurosecretory material by protrusions of bounding membranes extending through the axolemma,in Diphyllobothrium dendriticum (Cestoda)

Summary The release of neurosecretory material was studied in the cestode tapeworm Diphyllobothrium dendriticum. In plerocercoid larvae cultivated in vitro the neurosecretory vesicles release their content by way of protrusions extending through the axolemma into the intercellular space. The released material is directed predominantly towards the muscles. No omega figures were detected. The significance of this new type of release is discussed.The authors are greatly indebted to Mrs. Annukka Kärkkäinen and Mr. Esa Nummelin for technical assistance     

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.     

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.