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Chlorinated insecticides and PCBs in Asteroidea and Holothuroidea species from Rijeka Bay, Yugoslavia.

For an evaluation of the extent of pollution in Rijeka Bay, Yugoslavia by chlorinated hydrocarbons an investigation of levels in sediments and the most abundant benthic organisms (Asteroidea and Holothuroidea species) has been carried out. Samples were collected at several stations in October 1981. The concentrations of chlorinated insecticides and polychlorinated biphenyls (PCBs) in Asteroidea species were from 0.5 to 46.0 micrograms kg-1 for DDTtotal, and from 2.5 to 435 micrograms kg-1 (wet weight) for PCBs. For the PCBs in Holothuroidea species the values ranged from 4.4 to 37.2 micrograms kg-1 and for DDTtotal from 0.9 to 48.5 micrograms kg-1. The contamination of Asteroidea and Holothuroidea samples from Rijeka Bay with persistent chlorinated hydrocarbons was compared with other data for the same species from other parts of the world. Accumulation of chlorinated hydrocarbons by Asteroidea and Holothuroidea species from sediments of Rijeka Bay is discussed with respect to concentration factors (CF). The highest CF (70.0) (dry biota/dry sediment) was obtained for the intestines of Asteroidea species for PCBs and a value of only 2.6 was obtained for DDTtotal for the skin and muscle of Holothuroidea species. The relatively low bioaccumulation of PCBs and DDTs from sediments in the Rijeka Bay by the Asteroidea and Holothuroidea species is reassuring, because of the problem of dredging and ocean dumping of sediments polluted with persistent chlorinated hydrocarbons.

Animals↗

[Asteroidea, Echinoidea y Holothuroidea in shallow bottoms of Bahía de Loreto, Baja California Sur, Mexico].

To evaluate echinoderm distribution, abundance and density a double 50 m transect, with a side observation range of 2.5 m was used at each of 11 stations (bimonthly samplings, Aug. 1997-Febr. 1998). In Bahía de Loreto the Phylum Echinodermata consists of 26 taxa. The greatest mean abundance by transect and mean densities in order of importance for Echinoidea were: Echinometra vanbrunti (94.1 +/- 52.9 ind, 0.25 ind/m2), Centrostephanus coronatus (38.15 +/- 9.15 ind, 0.06 ind/m2), Tripneustes depressus (28.68 +/- 6.86 ind, 0.039 ind/m2), Eucidaris thouarsii (10.66 +/- 3.37 ind, 0.025 ind/m2) and Diadema mexicanum (11.75 +/- 4.92 ind, 0.023 ind/m2); for Asteroidea were: Phataria unifascialis, Mithrodia bradleyi and Acanthaster planci. Asteroidea was the dominant class with 12 species, followed by Echinoidea with ten and Holothuroidea was represented by four species.

Animals↗

Phylogenetic position of the enigmatic starfish family Podosphaerasteridae (Asteroidea, Valvatida) with a morphological observation of the skeletal structure by micro-CT.

Background The genus Podosphaeraster comprises seven species, characterised by a distinctive spherical body, all currently known from the seabed at depths below approximately 70 m. Its peculiar morphology has made its phylogenetic placement a subject of ongoing debate. It was initially suggested to be placed in Sphaerasteridae, the same family as fossil species. However, subsequent detailed skeletal analyses of the fossil forms suggested that this similarity was likely due to convergent evolution. Recent molecular analyses have revealed that Valvatida, in which the genus is currently placed, is likely a large polyphyletic group, leaving its taxonomic position still unresolved. New information A detailed examination of the internal skeletal structure of Podosphaeraster toyoshiomaruae, collected from the seas around Japan, was conducted using micro-focus X-ray computed tomography. Concurrently, shotgun sequencing was performed to identify key molecular markers for recent asteroid phylogeny. Additionally, shotgun sequencing determined the complete mitochondrial genome. Despite conservative evolution amongst asteroidean mitochondrial genomes, a translocation of the COX2 gene was revealed, representing the first discovery of a major protein-coding gene translocation within Asteroidea. In the phylogenetic tree, P. toyoshiomaruae was positioned as the most basal lineage within Valvatida. However, the statistical support for this placement was low, potentially due to the long-branch attraction caused by the excessively rapid evolutionary rate. Images reconstructed by micro-CT confirmed the presence of calcified reinforcement in the mesentery and showed its detailed structure for the first time. The mesentery skeleton was found to connect to the V-plate and five pairs of plates, including three kinds of marginal plates. This suggests that the marginal plates of this species may not be homologous with those of other asteroids. Although varying degrees of marginal plate reduction are shared with the order Velatida, we consider this to be a case of convergent evolution. Our phylogenetic analysis indicates a close relationship between P. toyoshiomaruae and Poraniidae (and other Valvatida), all of which possess marginal plates differentiated to varying extents, the homology of which remains uncertain.

Asteroidea↗

Comparative histological and immunohistochemical study of sea star tube feet (Echinodermata, Asteroidea).

Adhesion in sea stars is the function of specialized structures, the tube feet or podia, which are the external appendages of the water-vascular system. Adhesive secretions allow asteroid tube feet to perform multiple functions. Indeed, according to the sea star species considered, the tube feet may be involved in locomotion, fixation, or burrowing. Different tube foot shapes usually correspond to this variety of function. In this study, we investigated the variability of the morphology of sea star tube feet as well as the variability of the composition of their adhesive secretions. This second aspect was addressed by a comparative immunohistochemical study using antibodies raised against the adhesive material of the forcipulatid Asterias rubens. The tube feet from 14 sea star species representing five orders and 10 families of the Class Asteroidea were examined. The histological study revealed three main tube foot morphotypes, i.e., knob-ending, simple disc-ending, and reinforced disc-ending. Analysis of the results suggests that tube foot morphology is influenced by species habitat, but within limits imposed by the evolutionary lineage. In immunohistochemistry, on the other hand, the results were very homogeneous. In every species investigated there was a very strong immunolabeling of the adhesive cells, independently of the taxon considered, of the tube foot morphotype or function, or of the species habitat. This indicates that the adhesives in all the species considered are closely related, probably sharing many identical molecules or, at least, many identical epitopes on their constituents.

Animals↗

Amplification of ribosomal genes and formation of extrachromosomal nucleoli in oocytes of starfish Henricia hayashi (Asteroidea: Echinasteridae).

DNA and RNA specific dyes, Ag-NOR staining and in situ hybridization were used for studying the nucleolar apparatus in the growing oocytes of Henricia hayashi (Asteroidea: Echinasteridae). A plasmid containing ribosomal genes of Drosophila melanogaster (Kolchinsky et al., 1980) labelled with 3H by nick-translation served as an rDNA probe. Multiple extrachromosomal nucleoli are formed by the cascade type as a result of growth and subsequent fragmentation of the chromosomal (primary) rDNA body and its derivative extrachromosomal (secondary) rDNA bodies. Ribosomal genes were shown in all nucleolar structures. Argentophilia of the primary and secondary DNA bodies appears to be due to the dense packing of the rDNA-containing material. Ag(+) NORs were detected in the extrachromosomal multiple nucleoli and NOR complexes. Amplification of rDNA is a highly probable conclusion from the existing data.

Animals↗

Tauropine dehydrogenase from the starfish Asterina pectinifera (Echinodermata: Asteroidea): presence of opine production pathway in a deuterostome invertebrate.

Tauropine dehydrogenase (tauropine:NAD oxidoreductase; TaDH) was purified to homogeneity from the body wall of the starfish Asterina pectinifera Müller at Troschel(Echinodermata: Asteroidea) by means of (NH4)2SO4 precipitation followed by column chromatographies in DEAE-cellulose, Sephadex G75, Macro-prep ceramic hydroxyapatite, PBE 94, and Toyopearl HW50S. The enzyme was a monomeric protein of approximately 42000 Da and pI 5.2. The maximum rate of the tauropine biosynthetic reaction was observed at pH 6.0, and that of the tauropine catabolic reaction was at pH 8.7-9.2. Taurine and pyruvate were the preferred substrates. The tauropine catabolic reaction was inhibited by the substrate tauropine: the peak rate was observed at 12.5 mM. Apparent Km values for NADH, taurine, and pyruvate were 0.036 +/- 0.002, 21.3 +/- 1.6, and 0.46 +/- 0.02 mM, respectively, and for tauropine and NAD+ were 2.64 +/- 0.73 and 0.068 +/- 0.005 mM, respectively. The molecular and catalytic properties of the starfish TaDH were basically similar to those of TaDH from other species belonging to the lower invertebrate phyla and the middle phyla of Prostostomia. Tauropine accumulation in vivo during experimental anoxia was also demonstrated. These results gave clear evidence of opine production pathway in deutrostome invertebrate.

Amino Acid Oxidoreductases↗

Lectin histochemistry of the hyaline layer around the larvae of Patiriella species (Asteroidea) with different developmental modes.

Larvae of sea stars are surrounded by an extracellular matrix called the hyaline layer. The lectin-binding properties of this matrix were investigated in an ultrastructural study of Patiriella species having different modes of development. The planktonic bipinnaria and brachiolaria of P. regularis and the planktonic brachiolaria of P. calcar demonstrated the same labeling of the hyaline layer for three lectins: Con A, SBA, and WGA. In both species the outer coarse meshwork stained for all three lectins, whereas the intervillous layer displayed patchy labeling. In the benthic brachiolaria of P. exigua, the outer coarse meshwork displayed heavy labeling for all three lectins. The heavy labeling of the outer coarse meshwork of P. exigua compared with that of the other species suggests an increased number of lectin binding sites in the hyaline layer of this species. The similar ultrastructure and histochemistry of the hyaline layer of P. regularis and P. calcar may reflect similar requirements of their extracellular cover in their planktonic environment. Lectin labeling shows that hypertrophy of the hyaline layer of P. exigua, in particular the outer coarse meshwork, involves elaboration of the carbohydrate composition of the matrix. Modifications seen in the ultrastructure and histochemistry of the hyaline layer of P. exigua appear to be associated with the evolution of benthic development.

Animals↗

Mitochondrial rDNA phylogeny of the asteroidea suggests the primitiveness of the paxillosida.

Asteroids display four distinct modes of developmental patterns: the indirect mode, the nonbrachiolarian mode, the direct mode, and the mode with a barrel-shaped larva. Among them the former two are planktotrophic, whereas the latter two are lecithotrophic. The direct mode and the mode with a barrel-shaped larva are thought to have evolved from the more primitive planktotrophic mode, the nonbrachiolarian and the indirect mode, respectively. However, whether the nonbrachiolarian mode or the indirect mode is the more primitive in asteroids is unresolved, despite discussion since early this century. A key aspect of this problem is the phylogenetic status of paxillosidans, since the nonbrachiolarian mode and the mode with a barrel-shaped larva are seen only in paxillosidans. To resolve this problem, we performed a molecular phylogenetic study of asteroids, based on the nucleotide sequences of mitochondrial rDNAs. Phylogenetic trees support a close relationship between the Asterinidae and the Solasteridae. We suggest that the paxillosidans are not a monophyletic group; rather, the Luidiidae (one family of Paxillosida) is a sister group to the rest of the asteroids. Although some aspects of our results contradict a recent study by Lafay et al. (1995, Syst. Biol. 44: 190-208) based on 28S rRNA sequences, both studies agree on a paraphyletic nature for the paxillosidans. We conclude that characters shared by paxillosidans are primitive; hence the primitive mode of development in asteroids is the nonbrachiolarian mode.

Animals↗

Electron microscopy of extracellular materials during the development of a sea star, Patiria miniata (Echinodermata: Asteroidea).

The fine structure of conspicuous extracellular materials during the life history of a sea star (Patiria miniata) is described. The outer surface of the developing sea star is covered by two morphologically different cuticles that appear sequentially during ontogeny. The primary cuticle, which is about 120 nm thick and two-layered, is present from mid-blastula through the end of the larval stage. The secondary cuticle, which is about 1 micron thick and three-layered, first appears on the epidermis of the rudiment region of the larva and, after metamorphosis, covers the entire epidermis of the juvenile and adult stages. During ontogeny, there are only two conspicuous gut cuticles: the first lines the newly invaginated archenteron at the start of the gastrula stage, and the second lines the esophagus during the larval stage. A blastocoelic basal lamina first appears at mid-blastula and persists as subectodermal and subendodermal basal laminae. Ruthenium red-positive granules are detectable between the lateral surfaces of adjacent ectodermal cells during part of the gastrula stage; this transient intercellular material may possibly aid in lateral adhesion between cells.

Animals↗

Histofluorescence study and biochemical assay of catecholamines (dopamine and noradrenaline) during the course of arm-tip regeneration in the starfish, Asterina gibbosa (Echinodermata, Asteroidea).

In the starfish, Asterina gibbosa, the histofluorescence method gives evidence of an aminergic activity in the course of arm-tip regeneration. This activity can be detected as variations in the localization and intensity of a green fluorescence in the nerve structures of the arm, the radial nerve, and the circumoral nerve ring connected to it. Biochemical assays reveal that dopamine levels increase on the 2nd and 4th days of regeneration, when the blastema is forming and when differentiation commences. The level of noradrenaline also increases on the 2nd day after amputation of the arm.

Animals↗

First immunocytochemical study of echinoderm smooth muscle: the Antarctic cushionstar Odontaster validus Koehler (Echinodermata, Asteroidea).

Our immunocytochemical observations reveal that the muscle present in the tips of the arms of the Antarctic cushionstar Odontaster validus contains caldesmon and calponin but not troponin. Thus, the muscle clearly belongs to the smooth muscle category. Distributions of contractile proteins such as actin, myosin (the latter a typical vertebrate muscle filament protein), paramyosin, and miniparamyosin (the latter two being characteristic of thick invertebrate muscle filaments) were also determined immunocytochemically. The results suggest that the thin filaments of the starfish smooth muscle are similar to those of the vertebrate muscle, but that the thick filaments differ from those of vertebrates and possess traits that are also seen in the muscle organization of invertebrates. The absence from the O. validus muscle of titin and nebulin, proteins so far known almost exclusively from the striated vertebrate muscle, comes as no surprise, but immunoreactivity to mini-titin (a protein of the same family as titin and its replacement in invertebrates) was strong and unambiguously recognizable between filaments. Odontaster validus' histochemical characteristics may be a reflection of the phylogenetic position of the echinoderms as deuterostome invertebrates or they may express an adaptation of the muscle to the harsh environmental conditions under which it has to function in the Antarctic water.

Actins↗

Determination of polyamines in digestive and reproductive tissues of adult Asterias vulgaris (Echinodermata: Asteroidea).

1. The polyamines spermine, spermidine and putrescine were extracted from tissues of Asterias vulgaris and quantitated using thin layer chromatography. 2. In the pyloric caeca, mean (+/- SE) levels of spermine, spermidine and putrescine were 138(15), 86(24) and 415(77) nmol/g wet wt tissue, respectively. In the testes, levels were 95(12), 13(6) and less than 6 nmol/g wet wt. In the ovaries, levels were 105(9), 11(0.8) and 15(8) nmol/g wet wt. 3. High levels of polyamines in the pyloric caeca may be related to secretion or excretion in this tissue. 4. We hypothesize that polyamines will be important in the regulation of cellular activity in these tissues during the annual reproductive cycle.

Animals↗

Avoidance of crude-oil contaminated sediment by the Australian seastar, Patiriella exigua (Echinodermata: Asteroidea).

This study attempted to determine whether Patiriella exigua, an Australian seastar, could detect and/or avoid oiled sediment when given an equal choice of unoiled sediment. The sediment was spiked once to produce one of three concentrations of oiled sediment used in the test chambers versus unoiled sediment. Behavioral observations were repeated over a 32 day period to test the effects of aging the oiled sediment. Results show that Patiriella exigua was capable of detecting oiled sediment and/or an oiled environment. Seastars avoided oiled sediment, with significantly higher numbers choosing either to reside on the clean sediment (p<0.05) or to travel up the glass sides of the tanks (p<0.001). Avoidance of oiled sediment increased with increasing sediment oil concentrations. Aging the oiled sediment decreased the oil content of the sediment and increased the number of seastars able to inhabit it (p<0.001). A potential narcotic effect of exposure to oiled sediment was observed.

Animals↗

The phylogenetic status of Paxillosida (Asteroidea) based on complete mitochondrial DNA sequences.

One of the most important issues in asteroid phylogeny is the phylogenetic status of Paxillosida. This group lacks an anus and suckers on the tube feet in adults and does not develop the brachiolaria stage in early development. Two controversial hypotheses have been proposed for the phylogenetic status of Paxillosida, i.e., Paxillosida is primitive or rather specialized in asteroids. In this study, we determined the complete mitochondrial DNA nucleotide sequences from two paxillosidans (Astropecten polyacanthus and Luidia quinaria) and one forcipulatidan (Asterias amurensis). The mitochondrial genomes of the three asteroids were identical with respect to gene order and transcription direction, and were identical to the previously reported mitochondrial genomes of Asterina pectinifera (Valvatida) and Pisaster ochraceus (Forcipulatida) in this respect. Therefore, the comparison of genome structures was uninformative for the purposes of asteroid phylogeny. However, molecular phylogenetic analyses based on the amino acid sequences and the nucleotide sequences from the five asteroids supported the monophyly of the clade that included the two paxillosidans and Asterina. This suggests that the paxillosidan characters are secondarily derived ones.

Animals↗