Cell number and cellular composition in infusoriform larvae of dicyemid mesozoans (Phylum Dicyemida)

Cell numbers and cellular composition were examined in infusoriform larvae of 44 species of dicyemid mesozoans belonging to 6 genera; Conocyema, Dicyema, Dicyemennea, Dicyemodeca, Microcyema, and Pseudicyema. In addition, literature on infusoriform larvae of another 20 species was reviewed. Infusoriform larvae consist of a constant cell number which is species-specific. Small interspecific variations are found in total cell numbers, 35, 37, 39, 41 and 42. The most frequent cell number encountered in infusoriform larvae studied is either 37 or 39. Infusoriform larvae with 35 cells are found in three genera. Infusoriform larvae with 37 cells are found in four genera. Infusoriform larvae with 39 cells are found in four genera. Most differences in total cell numbers are due to the absence or presence of particular ventral cells. In all infusoriform larvae, the lateral, dorsal and caudal areas are cell constant, whereas in the apical and ventral areas a distinct and variable configuration of cells are present. In cellular composition, a total of 29 cells (15 cell types) were recognized in all infusoriform larvae examined. Additional cell types are characteristic of a relatively few species. Even in infusoriform larvae with the same total cell numbers, cellular composition varies by species. Thus, there are 7 variations of cellular composition in infusoriform larvae with 37 cells. Differences in larval cell numbers and types do not warrant traditional generic separation of dicyemids.     

Developmental patterns and cell lineages of vermiform embryos in dicyemid mesozoans.

Patterns of cell division and cell lineages of the vermiform embryos of dicyemid mesozoans were studied in four species belonging to four genera: Conocyema polymorpha, Dicyema apalachiensis, Microcyema vespa, and Pseudicyema nakaoi. During early development, the following common features were apparent: (1) the first cell division produces prospective cells that generate the anterior peripheral region of the embryo; (2) the second cell division produces prospective cells that generate the posterior peripheral region plus the internal cells of the embryo; (3) in the lineage of prospective internal cells, several divisions ultimately result in cell death of one of the daughter cells. Early developmental processes are almost identical in the vermiform embryos of all four dicyemid genera. The cell lineages appear to be invariant among embryos and are highly conserved among species. Species-specific differences appear during later stages of embryogenesis. The number of terminal divisions determines variations in peripheral cell numbers among genera and species. Thus, the numbers of peripheral cells are fixed and hence species-specific.     

Redescription of two Dicyemennea (Phylum: Dicyemida) from Rossia pacifica (Mollusca: Cephalopoda: Decapoda)

Two species of dicyemid mesozoan are redescribed from Rossia pacifica Berry, 1911, collected off Iwase in Toyama Bay, Honshu, Japan. Dicyemennea brevicephaloides Bogolepova-Dobrokhotova, 1962, is a large species that reaches about 4,000 microm in length. This species attaches to the surface of the branchial heart appendages. The vermiform stages are characterized as having 23 peripheral cells, a disc-shaped calotte, and an axial cell that extends to the propolar cells. An anterior abortive axial cell is absent in vermiform embryos. Infusoriform embryos consist of 35 cells; a single nucleus is present in each urn cell and the refringent bodies are absent. Dicyemennea rossiae Bogolepova-Dobrokhotova, 1962, is a medium species that reaches about 2000 microm in length. This species lives in folds of the renal appendages. The vermiform stages are characterized as having 29-34 peripheral cells, a conical calotte, and an axial cell that extends to the middle of the metapolar cells. An anterior abortive axial cell is present in vermiform embryos. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid.     

Three new species of dicyemid mesozoans (phylum Dicyemida) from Amphioctopus fangsiao (Mollusca: Cephalopoda), with comments on the occurrence patterns of dicyemids

Three new species of dicyemid mesozoans are described from the renal appendages of Amphioctopus fangsiao, collected off Akashi, in Harima Nada, and from Osaka Bay. Dicyema akashiense n. sp. is a small species that reaches about 900 microm in length. The vermiform stages are characterized as having 15-17 peripheral cells, a conical calotte, and an axial cell that extends to the base of the metapolar cells. Infusoriform embryos consist of 37 cells; two nuclei are present in each urn cell, and the refringent bodies are solid. Dicyema helocephalum n. sp. is a small species that reaches about 800 microm in length. The vermiform stages are characterized as having 22 peripheral cells, a disc-shaped calotte, and an axial cell that extends to the base of the propolar cells. Infusoriform embryos consist of 37 cells; a single nucleus is present in each urn cell, and the refringent bodies are solid. Dicyema awajiense n. sp. is a small species that reaches about 300 microm in length. The vermiform stages are characterized as having 22 peripheral cells, a conical calotte, and an axial cell that extends to the middle of the propolar cells. Infusoriform embryos consist of 37 cells; a single nucleus is present in each urn cell, and the refringent bodies are solid. In A. fangsiao various occurrence patterns of dicyemid species were observed, including instances where different dicyemid species were found in the renal appendage on each side. This suggests that dicyemids infect each renal appendage independently. The prevalence, reproductive traits, calotte shapes, and co-occurrence patterns of dicyemids are briefly discussed.     

Three new dicyemids from Octopus sasakii (Mollusca: Cephalopoda: Octopoda)

Three new species of dicyemid mesozoan are described from Octopus sasakii Taki, 1942, collected from Tosa Bay and Kii Strait in Japan. Dicyema shimantoense n. sp. is a medium-size species that reaches about 3,000 microm in length, and lives in folds of the renal appendages. The vermiform stages are characterized by having 22 peripheral cells, a conical calotte, and an axial cell that extends to the base of the propolar cells. Infusoriform embryos consist of 37 cells; a single nucleus is present in each urn cell, and the refringent bodies are solid. Dicyema codonocephalum n. sp. is also a medium-size species that reaches about 2,000 microm in length. It too lives in folds of the renal appendages. The vermiform stages are characterized by having 17-22 peripheral cells, an elongated calotte, and an axial cell that extends to the middle of propolar cells. Infusoriform embryos consist of 37 cells; a single nucleus is present in each urn cell, and the refringent bodies are solid. Dicyemennea pileum n. sp. is a medium species that reaches about 2,000 microm in length. It attaches to the surface of the renal appendages. The vermiform stages are characterized by having 23 peripheral cells, a disc-shaped calotte, and an axial cell that extends to the propolar cells. An anterior abortive axial cell is absent in vermiform embryos. Infusoriform embryos consist of 38 cells; 2 nuclei are present in each urn cell, and the refringent bodies are liquid. These are the first dicyemids to be described from Octopus sasakii.     

Redescription of Dicyemennea nouveli (Phylum: Dicyemida) from Enteroctopus dofleini (Mollusca: Cephalopoda: Octopoda)

A species of dicyemid mesozoan is redescribed from the giant Pacific octopus, Enteroctopus dofleini (Wülker, 1910), collected off Iwase in Toyama Bay, Honshu, Japan. Dicyemennea nouveli McConnaughey, 1959, is a large species that reaches about 12,000 microm in length. This species lives in folds of the renal appendages. The vermiform stages are characterized as having 30-41 peripheral cells, a conical calotte, and an axial cell that extends to the middle of the metapolar cells. An anterior abortive axial cell is present in vermiform embryos. Full-grown vermiform embryos have as many as 4 agametes. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid.     

Fine structure of dicyemid mesozoans,with special reference to cell junctions

The fine structure of the dicyemid mesozoan, Dicyema acuticephalum, from Octopus vulgaris, was studied with special attention to intercellular junctional complexes between various kinds of cells. Two types of intercellular junction, namely, adherens junctions and gap junctions, were found in both vermiform stages and in infusoriform embryos. Adherens junctions were classified into two types. Zonulae adherentes-like junctions were observed between adjacent peripheral cells at vermiform stages, between adjacent external cells of infusoriform embryos, and between members of groups of internal cells that covered the urn in infusoriform embryos. Maculae adherentes-like junctions were seen between a peripheral cell and an axial cell at vermiform stages. In infusoriform embryos, these junctions were observed between various types of cells, excluding urn cells. Gap junctions were found between adjacent peripheral cells at vermiform stages, whereas in infusoriform embryos these junctions were located between various types of cells excluding urn cells. Dicyemids might be the most primitive multicellular animals to possess these basic types of cell junctions. Ciliary rootlet systems at vermiform stages and in infusoriform embryos were unique in structure compared with those of other primitive multicellular animals. J Morphol 231:297–305, 1997. © 1997 Wiley-Liss, Inc.     

New species of Dicyemennea (phylum: Dicyemida) in deep-water Graneledone (Mollusca: Cephalopoda: Octopoda) from the Antarctic

Two new species of dicyemids are described from 2 species of deep benthic cephalopods, Graneledone antarctica and G. macrotyla, collected in the Southern Ocean south of the Antarctic Convergence. Dicyemennea bathybenthum n. sp. was found in G. antarctica. It is a medium-sized dicyemid whose length does not exceed 1,000 microm. The calotte is bluntly rounded or conical in small individuals, but the shape becomes discoidal in large individuals. Nematogens and vermiform embryos have 23 peripheral cells. An anterior abortive axial cell is present in vermiform embryos. Other stages in the life cycle were not observed. Dicyemennea dorycephalum n. sp. was found in G. macrotyla. It is a medium to large dicyemid that rarely exceeds 4,000 microm. The calotte is distinctly pointed, similar in shape to a spearhead. Vermiform stages typically have either 25 or 27 peripheral cells. An anterior abortive axial cell is present in vermiform embryos. Infusoriform embryos have 37 cells, the refringent bodies are solid in composition, and 2 nuclei are present in each urn cell.     

A new dicyemid from Benthoctopus sibiricus (Mollusca: Cephalopoda: Octopoda)

A new species of dicyemid mesozoan is described from Benthoctopus sibiricus Loyning, 1930, collected in the eastern Chukchi Sea. Dicyemennea chukchiense n. sp. is a medium species that reaches about 2,000 μm in length; it lives in folds of the renal appendages. The vermiform stages are characterized in having 23 peripheral cells, a conical calotte, and an axial cell that extends to the base of the propolar cells. An anterior abortive axial cell is present in vermiform embryos. Infusoriform embryos consist of 37 cells; a single nucleus is present in each urn cell and the refringent bodies are solid. This is the first dicyemid described from the Arctic waters.     

Dicyema shorti n. sp. (phylum Dicyemida) from Octopus burryi (Mollusca: Cephalopoda: Octopodidae) in the Gulf of Mexico

Nematogens and vermiform embryos of a new species of Dicyema are described from an octopus collected off Veracruz, Mexico. Dicyema shorti n. sp. is a small dicyemid species that rarely exceeds 500 microm in length. It is further characterized by the presence of 18 peripheral cells in the vermiform stages, a conical-shape calotte, and an axial cell that extends to the base of the propolar cells. Other stages in the life cycle of the parasite are not known. This is the first dicyemid to be described from Octopus burryi Voss. 1950, and also from both the southern Gulf of Mexico and the country of Mexico.     

Serotonin immunoreactivity in a highly enigmatic metazoan phylum, the pre-nervous Dicyemida

Dicyemida are simply organized metazoans with a long axial cell surrounded by a layer of ciliated peripheral cells. Stem cells and propagates develop inside the cytoplasm of the axial cell. Two types of propagates exist, asexually formed vermiform embryos and sexually derived infusoriform larvae, neither of which, however, develop by gastrulation. Thus, Dicyemida lack all traits typical of common metazoa and represent a pre-nervous state of organization. In this study, immunofluorescence and immunoelectron microscopy have been used to investigate the presence of a serotonin-like molecule in adults and larvae of Dicyema typus and Dicyemennea antarcticensis. Generally, in the adults of both species, the main immunoreactive structures are confined either to small vesicles inside the peripheral cells or to prominent vesicles in the axial cell, the latter vesicles apparently playing a role during the differentiation of the stem cells and the development of the propagates. In all stages, faint immunoreactivity can be observed over the surface membrane and the ciliary apparatus and over the heterochromatic blocks in the nuclei. Surprisingly, in the infusoriform larva, striking immunoreactivity occurs in the vesicles of the capsule cells covering the urn cells, which contain germinal cells. These vesicles might represent stores used in a later life phase of the animal. The results suggest that, in the pre-nervous Dicyemida, serotonin in its ancestral form may function as a hormone regulating essential developmental and morphogenetic processes.     

Epibolic extension of the presumptive ectodermal layer of embryos of the newt Cynops pyrrhogaster before and during gastrulation.

Epibolic extension of the presumptive ectodermal layer (PEL) was investigated in embryos of the newt Cynops pyrrhogaster before and during gastrulation. The PEL was composed of only one layer of columnar cells at all stages examined. The cells of the PEL became elongated from the blastula to the early gastrula stage. They were most elongated at the early gastrula stage and then shortened during gastrulation. Present observations suggest that changes in cell shape of the PEL play an important role in the control of the epibolic extension of the newt embryos. The morphology and movement of the isolated cells from the PEL were examined in an attempt to elucidate the role of cell movement in epibolic extension of the PEL. Blebbing and vermiform cells which showed active cell movement appeared at the early blastula stage. The blebbing cells, which formed large hyaline blebs that moved around the circumference of each cell, appeared in large numbers at the early blastula stage. The frequency of the blebbing cells decreased from the early blastula to the early gastrula stage and increased again during gastrulation. The vermiform cells, which had an elongated cell body and moved in a worm-like manner, increased in frequency from the early blastula to the early gastrula stage. The relative number of such vermiform cells was maximal at the early gastrula stage and decreased abruptly during gastrulation. These results suggest that the elongation of the cells of the PEL is controlled by the active cell movement which resembles that of a worm.     

Survival and differential development of Entomophaga maimaiga and Entomophaga aulicae (Zygomycetes: Entomophthorales) in Lymantria dispar hemolymph

The closely related entomophthoralean fungi Entomophaga aulicae and E. maimaiga are both host-specific pathogens of lepidopteran larvae. However, these fungi do not have the same host range. The first objective of this study was to compare the fate of E. aulicae in the nonpermissive host Lymantria dispar with the fate of the successful pathogen E. maimaiga over the same time period. In the hemolymph of L. dispar injected with E. maimaiga protoplasts, the number of hemocytes demonstrated a decreasing trend after the first day postinjection and hemocytes completely disappeared by day 5, with the majority of larvae dying in 5.6 +/- 0.1 days. In L. dispar larvae, E. maimaiga infections developed successfully, evidenced by increasing numbers of protoplasts and hyphal bodies prior to host mortality. In contrast, at day 5 hemocytes were readily visible in hemolymph of E. aulicae-injected larvae, but E. aulicae cells did not increase in numbers, although persisting in the hemolymph for at least 16 days postinjection. For both fungal species, when hemolymph samples from injected insects were introduced to culture media viable fungal cultures were always produced. Both E. aulicae and E. maimaiga occurred in hemolymph initially after injection as protoplasts. For E. maimaiga, after day 3, <50% of fungal cells were hyphal bodies until insect death when most cells regenerated cell walls. For E. aulicae, from day 2 equal numbers of fungal cells in the hemolymph occurred as protoplasts and hyphal bodies. To investigate the cause of fungistasis in E. aulicae-injected larvae, E. aulicae cell cultures exposed to partially purified protein fractions from hemolymph of larvae infected with either fungus displayed increased lysis and decreased viability at lower concentrations of protein fractions compared with E. maimaiga cell cultures. These studies demonstrate that E. aulicae does not increase in L. dispar hemolymph, although it persists and results suggest that proteinaceous factors induced within the hemolymph may limit the capacity of E. aulicae to develop successful infections.     

Dicyemennea canadensis n. sp. (phylum Dicyemida) from Bathypolypus arcticus (Mollusca: Cephalopoda: Octopoda)

Dicyemennea canadensis n. sp. is described from a bathyal octopus collected off Canada in the Bay of Fundy. The dicyemid is a small species that rarely exceeds 600 microm in length. The vermiform stages are further characterized as having 17-23 peripheral cells, a conical-shaped calotte, an axial cell that extends to the base of the propoplar cells, and no abortive axial cells. Infusoriform embryos consist of 37 cells. There is 1 nucleus in each urn cell, and refringent bodies are absent. This is the first dicyemid to be described from the cephalopod Bathypolypus arcticus (Prosch. 1847), and the first dicyemid reported from Canada. In addition, it is the first species of Dicyemennea from the northwestern Atlantic Ocean to be described.     

The control of chloroplast number in wheat mesophyll cells

Chloroplast number per cell and mesophyll cell plan area were determined in populations of separated cells from the primary leaves of different wheat species representing three levels of ploidy. Mean chloroplast number per cell increases with ploidy level as mean cell size increases. But in addition the analysis of individual cells clearly shows that cells of a similar size but from species of different ploidies have similar numbers of chloroplasts. We conclude that the number of chloroplasts within a cell is closely correlated (P<0.001) with the size of the cell and this relationship is consistent for species of different ploidies over a wide range of cell sizes. These results are discussed in relation to the hypothesis that chloroplast number in leaf mesophyll cells is determined by the size of the cell.     

Suppression of Nippostrongylus brasiliensis (Nematoda)-induced lysophospholipase activity and peripheral eosinophilia by Eimeria nieschulzi (Apicomplexa)

Interspecific interactions between Nippostrongylus brasiliensis and Eimeria nieschulzi were studied by measuring fecal lysophospholipase (LYPH) activity and relative numbers of peripheral eosinophils in rats singly or concurrently infected with one or both parasite species. Three groups of 10 rats each were inoculated with 2 X 10(3) N. brasiliensis L3 larvae and/or 5 X 10(5) E. nieschulzi sporulated oocysts. Groups 1 and 2 were infected with E. nieschulzi or N. brasiliensis, respectively. Group 3 rats were infected first with N. brasiliensis, followed on day 8 postinoculation (PI) with E. nieschulzi. Each rat served as its own control. Results revealed LYPH levels rose steadily in Group 2 rats, reaching significant peaks on days 10 and 12 PI before decreasing to control levels. Lysophospholipase activity in Groups 1 and 3, however, did not differ from control values. Group 2 rats also demonstrated peripheral eosinophilia, with peak values occurring on days 10, 12, 14, and 16 PI, while rats in Groups 1 and 3 exhibited no eosinophilia. These results demonstrate that E. nieschulzi suppressed intestinal LYPH activity and relative peripheral eosinophilia and demonstrate that a host's immune response to a single parasite may be significantly altered when a second parasite species is present.     

Light and electron microscopy studies of the ulnar, saphenous, and caudal cutaneous sural nerves of the dog

Transverse sections of the ulnar, saphenous, and sural nerves taken at specific levels in normal, young-adult beagle dogs were examined qualitatively and quantitatively at both the light and electron microscopic levels. The aim of this investigation was to provide baseline information for future studies of peripheral nerve disease in this species. A systematic sampling technique was used for the determination of nerve components (i.e., unmyelinated axons and Schwann cell and fibroblast nuclei). In all nerves sampled, the average size distribution for unmyelinated axons was unimodal, and most of the axons were 0.4-1.1 micron in diameter. Within this range, there were slight individual and nerve-to-nerve variations in the location of the largest diameter peak. The mean densities of Schwann cell nuclei (numbers/mm2) ranged from 841/mm2 in the palmar branch of the ulnar nerve, to 1,223/mm2 in the caudal cutaneous sural nerve, being nearly four times the average density or fibroblast nuclei. In every animal and in almost every nerve, a few abnormalities were found; and these should be kept in mind when assessing peripheral nerves in the dog.     

Spontaneous micronuclei in peripheral blood erythrocytes from 54 animal species (mammals, reptiles and birds): part two

The normal numbers of micronucleated erythrocytes (MNE) observed in peripheral blood samples differ among species. This depends on the effectiveness of the spleen (or the rest of the reticuloendothelial system) to withdraw them from circulation. In our previous report, we assessed the number of MNE in the peripheral blood of 35 mammalian species. Here we show the results observed in 54 species including mammals, reptiles and birds. We obtained 212 peripheral blood samples from different species. In 14 species, only one individual was studied. Slides were stained with acridine orange. The total number of MNE (normo and polychromatic) in 10,000 erythrocytes per animal are shown. The species that display the higher MNE were: ocelote, lynx, owl, gray squirrel, hedgehog, lion, orange fronted parakeet and common barn owl. For this reason, these species could be tested as monitors for genotoxic events. Another interesting observation was that in the gray squirrel, we found the highest values of MNE in the smaller (younger) animals when compared with the larger (older) of the same species.