A chromatophore system in the hermatypic,deep-water coral Leptoseris fragilis (Anthozoa: Hexacorallia)

Ultrastructural evidence is presented of a chromatophoresystem in the zooxanthellae containing hermatypic, deep-water coral Leptoseris fragilis (Milne Edwards and Haime). It consists of multilobed cells which mainly occupy the intercellular space of the oral gastrodermis. The cellular processes are filled with electron-dense granules up to 1-m-long and 0.5-m-wide. Within the cytoplasm an elaborate system of microtubules is established. The ramifications of the pigment cells, containing the pigment granules, form a dense and nearly continuous layer close to the overlying zooxanthellae. It is speculated that host pigments may transform the violet portion of the incident light into longer wavelengths, thus increasing the photosynthetic efficiency of the zooxanthellae.     

Photosynthesis and growth of red and green morphotypes of Kappaphycus alvarezii (Rhodophyta) from the Philippines

The effect of photosynthetic available radiation (PAR) levels, light quality, ultraviolet (UV) radiation, and temperature on photosynthesis, growth, and chlorophyll fluorescence was evaluated in red and green morphotypes of the rhodophyte Kappaphycus alvarezii (Doty) Doty under controlled conditions. Chlorophyll a and phycoerythrin (PE) levels were similar in the red and green morphotypes cultured under the same conditions, but phycocyanin (PC) and allophycocyanin (APC) levels were 2-fold greater in the green than in the red morphotype. Pigment characterization indicated that the overexpression of PC and APC masked the red pigmentation in the green morphotype. Maximum photosynthesis and photosynthetic efficiency were similar between the two morphotypes assayed at a wide temperature range, which was reflected in the similar growth rates observed in outdoor culture systems. In the green morphotype, photosynthetic efficiency increased 2-fold relative to the red morphotype when assayed with red light (λ > 600 nm), indicating that photosynthetic characteristics are modified as a result of pigment variation in these morphotypes. Such increase in photosynthetic efficiency in the green morphotype, however, did not result in greater growth rates when cultured under white light. Short exposure to high levels of solar radiation (UV-A + UV-B + PAR), and filtered solar radiation (UV-A + PAR or PAR) decreased effective quantum yield (ΔF/F _m′) in both morphotypes. The reduction of ΔF/F _m′ values in the red and green morphotypes was accounted for by high levels of PAR and not by the UV-A + UV-B + PAR and UV-A + PAR treatments. Photoinhibition caused by UV-A, UV-B, or PAR was completely reversed within 30 h after incubations. Recovery rates from photoinhibition, however, were significantly reduced in the green morphotype when incubated with UV-B radiation. The results here suggest that the overexpression of pigments do not necessarily increase photosynthesis and growth in these morphotypes. Received: 19 June 2000 / Accepted: 28 November 2000     

Photobiology of endolithic microorganisms in living coral skeletons: 1. Pigmentation, spectral reflectance and variable chlorophyll fluorescence analysis of endoliths in the massive corals Cyphastrea serailia, Porites lutea and Goniastrea australensis

We used microscopy, reflectance spectroscopy, pigment analysis, and photosynthesis-irradiance curves measured with variable fluorescence techniques to characterise the endolithic communities of phototrophic microorganisms in the skeleton of three massive corals from a shallow reef flat. Microscopic observations and reflectance spectra showed the presence of up to four distinct bands of photosynthetic microorganisms at different depths within the coral skeleton. Endolithic communities closer to the coral surface exhibited higher photosynthetic electron transport rates and a green zone dominated by Ostreobium quekettii nearest the surface had the greatest chlorophyll pigment concentration. However, Ostreobium was also present and photosynthetically active in the colourless band between the coral tissue and the green band. The spectral properties and pigment density of the endolithic bands were also found to closely correlate to photosynthetic rates as assessed by fluorometry. All endolithic communities were extremely shade-adapted, and photosynthesis was saturated at irradiances <7 μmol photons m⁻²s⁻¹.     

Different responses of photosynthesis and flow cytometric signals to iron limitation and nitrogen source in coastal and oceanic Synechococcus strains (Cyanophyceae)

Iron plays an important role in marine primary productivity, and Synechococcus species as major contributors to the total photosynthetic biomass in the world’s oceans might be limited by iron supply in some regions. The present study aimed to compare the photosynthesis and flow cytometric signals of four Synechococcus strains grown under different iron concentrations with either nitrate or ammonium as the sole nitrogen source. Two oceanic strains were much more sensitive to iron limitation than two coastal strains. The inhibition of iron limitation on the growth, maximal PSII photochemical yield, maximal rate of relative electron transport and photochemical quenching of the two oceanic strains was higher than for their coastal counterparts. Under iron limitation condition, the connectivity factor between individual photosynthetic units (ρ) increased for the two coastal strains, while decreased for the two oceanic strains. Furthermore, iron limitation accelerated the Q _A re-oxidation of the two oceanic strains and the PQ pool re-oxidation of the two coastal strains. Under iron limitation condition, the cell size of the two coastal strains and intracellular pigment concentrations of the two oceanic strains decreased, while the side light scatter/front light scatter (SS/FS) ratio of the two coastal strains increased. In contrast to iron limitation, nitrogen source only marginally affected the photosynthesis of the four Synechococcus strains. Ammonium enhanced the growth of the two coastal strains under iron-replete condition. For the two oceanic strains, ammonium increased their cell size and decreased their SS/FS ratio and intracellular pigment concentrations under iron-deplete and iron-replete conditions.     

Lysosomes and sulfide-oxidizing bodies in the bacteriocytes of Lucina pectinata, a cytochemical and microanalysis approach

Lucina pectinata is a large tropical clam living deeply burrowed in the black, reducing mud of mangrove swamps. It is known to possess hemoglobin in the cytoplasmic areas of its bacteriocytes, which harbor sulfide-oxidizing bacteria. The bacteriocytes also possess lysosome-like microbodies containing either membrane whorls or electron-dense granules in which free heme compounds have been identified. The cytochemical detection of acid phosphatase and arylsulphatase through EDX (energy-dispersive X-ray) microanalysis strongly suggests that the bacteriocytes of L. pectinata contain, in fact, two different types of microbodies. Some of these (devoid of dense granules) possess a variable amount of lysosomal enzymes and occasionally a limited quantity of iron, which may result from a recycling process of hemoglobin. Their main function seems to be the digestion of a limited proportion of symbiotic bacteria. They represent genuine secondary lysosomes with a functionally acidic pH. The second type of microbodies is characterized by dense granules containing sulfur and iron hemes but no lysosomal enzymes. Their sulfide-oxidizing activity was substantiated by benzyl viologen assay, with Na₂S as a substrate. These microbodies appear to be similar to the sulfide-oxidizing bodies (SOBs) described in the bacteriocytes of other bivalve species with symbiotic thioautotrophic bacteria; however, their sulfide-oxidizing activity appears to be non-enzymatic. They are discrete organelles, characterized by a functionally basic pH and pseudoperoxidasic activity, and have been termed SOBs. Therefore, the bacteriocytes of L. pectinata possess at the same time functional lysosomes and functional SOBs. Received: 17 August 2000 / Accepted: 20 December 2000     

Degrees of vacuolation of the absorptive intestinal cells of five Sagitta (Chaetognatha) species: possible ecophysiological implications

The cell organization and ultrastructure of the intestine have been studied in five species of Sagitta, one epiplanktonic (S. bipunctata Quoy and Gaimard, 1827), four mesoplanktonic (S. megalophthalma Dallot and Ducret, 1969, S. decipiens Fowler, 1905, S. minima Grassi, 1881, S. zetesios Fowler, 1905). The intestinal epithelium is composed of two ciliated cell types. The first (S-cell) principally occurs in the anterior intestine and represents typical secretory cells. The second (A-cell) occurs in the median and posterior intestine and displays ultrastructural features involved in absorption and intracellular digestion, that is, coated pits, endocytotic vesicles, cytoplasmic tubules, and two distinct types of digestive vacuoles. Whereas S-cells exhibit few ultrastructural differences among the five species, the vacuolar volume of the A-cells located in the mid portion of the intestine is higher in the mesoplanktonic species. In S. zetesios and S. megalophthalma, each side of the median intestine comprises several hypervacuolated A-cells visible in transverse section; their degree of vacuolation is inversely proportional to their number. The increased volume of the intestinal vacuoles is most marked in S. decipiens and S. minima; in both these species, each lateral side of the median intestine displays a single ultravacuolated A-cell. The possible ecophysiological implications of intestinal vacuoles are discussed in relation to patterns of the vertical distribution of several Sagitta species. The vacuoles are presumed to regulate buoyancy, enabling the mesoplanktonic species to make vertical diel migrations. Received: 22 January 2000 / Accepted: 28 July 2000     

Deleterious effects of a nonPST bioactive compound(s) from <Emphasis Type="Italic">Alexandrium tamarense</Emphasis> on bivalve hemocytes

The known negative effects of shellfish toxin-producing dinoflagellates on feeding, burrowing and survival of some bivalve mollusks has prompted questions concerning whether they might also impair the internal defense system of affected bivalves and make them more susceptible to disease agents. The primary components of the cellular defense system are hemocytes. Many toxic dinoflagellates are too large to be ingested whole by hemocytes and would most likely be exposed to intracellular toxins only after the algae are consumed, broken down, and the water-soluble toxins, released. Therefore, we conducted a series of experiments in which hemocytes from two suspension-feeding bivalves—the Manila clam, Ruditapes philippinarum, and the softshell clam, Mya arenaria—were exposed in vitro to filtered extracts of one highly toxic paralytic shellfish toxin (PST)-producing and one nonPST-producing strain of Alexandrium tamarense (isolates PR18b, 76 ± 6 STXeq cell⁻¹ and CCMP115, with undetectable PST, respectively). We measured adherence and phagocytosis, two hemocyte attributes known to be inhibited by bacterial pathogens and other stressors. We found no measurable effect of a cell-free extract from a highly concentrated suspension of the PST-producing strain on hemocytes of either bivalve species. Instead, extract from the nonPST-producing strain had a consistent negative effect on both clams, resulting in significantly lower adherence and phagocytosis compared to strain PR18b and filtered seawater controls. The bioactive compound produced by strain CCMP115, which has yet to be characterized, may be similar to the PST-independent allelopathic compounds described for Alexandrium spp., which act on other plankters. These compounds and those produced by other harmful algae are known to cause immobilization, cellular deformation and lysis of co-occurring target organisms. Thus, nonPST producing Alexandrium spp., which do not cause paralysis and burrowing incapacitation of clams, may still produce a compound(s) that has negative effects not only on hemocytes, but on other molluscan cell types and their functions, as well.     

Photoadaptation of zooxanthellae in the sponge Cliona vastifica from the Red Sea, as measured in situ

In the Red Sea, the zooxanthellate sponge Cliona vastifica (Hancock) is mainly present at >15 m depth or in shaded areas. To test whether its scarcity in unshaded areas of shallower waters is linked to the functional inefficiency of its photosymbionts at high irradiances, sponges were transferred from 30 m to a six times higher light regime at 12 m depth, and then returned to their original location. During this time, photosynthetic responses to irradiance were measured as rapid light curves (RLCs) in situ by pulse amplitude modulated (PAM) fluorometry using a portable underwater device, and samples were taken for microscopic determinations of zooxanthellar abundance. The zooxanthellae harboured by this sponge adapted to the higher irradiance at 12 m by increasing both their light saturation points and relative photosynthetic electron transport rates (ETRs). The ETRs at light saturation increased almost fourfold within 15–20 days of transfer to the shallower water, and decreased back to almost their original values after the sponges were returned to 30 m depth. This, as well as the fact that the photosynthetic light responses within an individual sponge were in accordance with the irradiance incident to specific surfaces, shows that these photosymbionts are highly adaptable to various irradiances. There was no significant change in the number of zooxanthellae per sponge area throughout these experiments, and the different photosynthetic responses were likely due to adaptations of the photosynthetic apparatus within each zooxanthella. In conclusion, it seems that parameters other than the hypothesised inability of the photosymbionts to adapt adequately to high light conditions are the cause of C. vastifica's rareness in unshaded shallow areas of the Red Sea. Received: 25 April 2000 / Accepted: 13 October 2000     

Photosynthetic activity of the non-dormant <Emphasis Type="Italic">Posidonia oceanica</Emphasis> seed

The photosynthetic adaptive features of non-dormant seeds in Posidonia oceanica were studied in order to evaluate the effects of light on germination success. Transmission electron micrographs showed the presence of chloroplasts in the epidermal cells, close to the nucleus at the periphery of the cytoplasm. The well-developed thylakoid membranes and the presence of starch granules indicated that the chloroplasts were photosynthetically active. The relationship between photosynthesis versus irradiance in P. oceanica seeds incubated at 15 and 21°C was analysed. The net photosynthesis in the non-dormant seed of P. oceanica was positive and compensated its respiration demand (90 μmol quanta m⁻² s⁻¹) at both temperatures. Net photosynthesis was negative at the other irradiance values. To test the effects of light on germination success, seeds were placed both in dark and light conditions. Germination success was significantly higher in light rather than in dark condition. The characteristics observed in the photosynthesis in P. oceanica seed could be a mechanism to guarantee seedling survival in temperate waters, demonstrating though the specialized nature of this species.     

Dinoflagellate <Emphasis Type="Italic">Alexandrium leei</Emphasis> (Dinophyceae) from Singapore coastal waters produces a water-soluble ichthyotoxin

A strain of Alexandrium leei Balech that was isolated in October 2002 from Singapore coastal waters and identified by light and scanning electron microscopy and phylogenetic analysis using LSU rDNA sequences, is toxic to Asian sea bass fingerlings (Lates calcarifer Bloch). The ichthyotoxicity of the algal cells obtained by filtration (15 μm mesh net) and rinsed with sterile culture medium indicated that the toxicity of A. leei was probably not due to bacterial contamination, which was further supported by a negative correlation between the time to death of the fish and the dosage of algal cells applied. Paralytic shellfish toxins (PST) could not be detected indicating that PST was not the cause of fish mortality. Fish bioassays using frozen culture, heat-treated cultures, cell-free culture medium, and hexane, ethyl acetate, and water extracts of algal cells indicated that A. leei produces a heat-stable, polar ichthyotoxin(s) which can be released from the algal cells into the culture medium. Phylogenetic analysis based on the LSU rDNA sequences of A. leei confirmed its identification and indicated that the Singapore strain is more similar to isolates from Malaysia than to a geographically distant strain from Korea. This is the first evidence of icthyotoxin production by A. leei.     

Fine structure of naive and allogeneic challenged ampullae in Botrylloides subpopulation I from the Mediterranean coast of Israel

The external peripheral termini of vasculature in botryllid ascidians (ampullae) are important in the expression of effector mechanisms during histocompatibility reactions. We study here morphological changes to the microstructure during allorecognition in Botrylloides subpopulation I (SP1) from the Mediterranean coast of Israel, where special giant ampullae, called megaloampullae are developed hours after first allogeneic challenge. The basal part of each naive ampulla consists of a sheath of continuous squamous epithelial cells (1 μm×20 μm). At the tip of the ampullae, about 130 epithelial cells become columnar with wide apical pads (15 μm×5–10 μm), that contain electron-opaque “adhesive vesicles”. Foci of crystallizations were observed within the vesicles of some columnar cells. Ampullar epithelial cells develop internal folds that support tight attachments between circulating blood cells (most of them are pigment cells) and ampullar epithelium. During the histoincompatibility process, the tunic peripheral edge at the interaction area becomes highly convoluted. Filaments (up to 1.16 μm) accumulate in the interaction tunic matrix. Some of these filaments encircle destructed cells. Fragmented cell membranes are also found next to the reacting ampullae. The most characteristic feature of the Botrylloides SP1 rejection reaction is dilation of the ampullae. The ampullar epithelium becomes “curly” when megaloampullae are formed. Within the curly region, about 30% of the squamous epithelial cells are further stretched, up to 33 μm in length. Some additional mechanisms are suggested to explain the fast increase (up to seven times) in ampullar width that is recorded within a few hours from the first allogeneic contact.     

Fine structure of some prosobranch ospharadia

The fine structure of the osphradia of Buccinum undatum 1. three Hawaiian Conus spp., Nassarius reticulatus (L), Nucella lapillus (L) and Littorina littorea (L) was examined. There is a remarkable uniformity in the arrangement of ultrastructural cell types in all the ospharadia investigated. The marginal glandular region is characterised by a single layer of cells, bearing a regular pallisade of microvilli and including two types of mucous cell. The epithelium of the sensory region is several cells deep, and bears a complex layer of microvilli in many orientations. Besides indifferent cells it contains two types of cell bodies of ciliated nerve processes. One type ends at the surface of the sensory region. The other, containing an elaborate array of smooth membranes in the perikaryon, has a process which enters the transitional region between sensory and glandular areas. In the transitional region, specialised cells with motile cilia adjoin the glandular region, and similar, but unciliated cells, containing large pigment granules scattered throughout the cytoplasm, adjoin the sensory region. The distal surface of both cell types is dissected by elefts extending 2 to 3 proximally, then widening into extensive spaces which contain cytoplasmic processes. The ciliated neurites which enter the transitional region end in the spaces at the base of the unciliated transitional cell elefts. It is suggested that the neuroepithelial cell processes, both those distributed over the sensory region and those concentrated in the transitional region, are receptors.     

An electron-microscope study of periostracum formation in some marine bivalves. II. The cells lining the periostracal groove

The histology and ultrastructure of the epithelial cells of the outer and middle mantle folds of the bivalves Mytilus edulis (L), Cardium edule (L), Macoma balthica (L) and Nucula sulcata Bronn are described. The cells lining the inner face of the outer fold exhibit a prominent granular endoplasmic reticulum, and Golgi complexes which are concerned with the elaboration of granules and vesicles eventually incorporated into the periostracum. A gradual reduction in the protein synthetic apparatus occurs towards the tip of the fold. Within the cells, it is proposed that the ovoid inclusion bodies are lysosomes and that they control the rate of secretion. The cells of the middle fold are cuboidal in appearance. Those of M. edulis and N. sulcata exhibit prominent granules, whereas those of C. edule and M. balthica possess vesicles. The cells of M. edulis differ from the others in possessing stout bundles of filaments, which occupy large areas of the cell and constitute a cell web. The cells of the epithelium in all cases do not appear to be implicated in periostracum formation.     

Population differences in nerve resistance to paralytic shellfish toxins in softshell clam, <Emphasis Type="Italic">Mya arenaria</Emphasis>, associated with sodium channel mutations

The softshell clam, Mya arenaria, is a commercially important bivalve with wide latitudinal distribution in North America. Populations of clams with a history of repeated exposure to toxic Alexandrium spp. have developed a natural resistance to the paralytic shellfish toxins (PSTs) produced by these algae. An association between PST resistance in individual clams and a single mutation in the saxitoxin (STX) binding region of the α-subunit of the voltage-gated sodium (Na⁺) channel gene was previously identified. Here we establish that more than one mutation associated with nerve resistance to STX occurred at this locus. Both cDNA from mRNA and genomic DNA sequences from individual clams are identical demonstrating that both alleles are expressed simultaneously. In addition, one resistant allele per individual is sufficient to confer neural resistance to STX even though heterozygous individuals show an intermediate level of resistance to STX in in vitro nerve trunk assays.     

Presence of Alexandrium catenella and paralytic shellfish toxins in finfish, shellfish and rock crabs in Monterey Bay, California, USA

The central California coast is a highly productive, biodiverse region that is frequently affected by the toxin-producing dinoflagellate Alexandrium catenella. Despite the consistent presence of A. catenella along our coast, very little is known about the movement of its toxins through local marine food webs. In the present study, we investigated 13 species of commercial finfish and rock crabs harvested in Monterey Bay, California for the presence of paralytic shellfish toxins (PSTs) and compared them to the presence of A. catenella and PSTs in sentinel shellfish over a 3-year period. Between 2003 and 2005, A. catenella was noted in 55% of surface water samples (n = 307) and reached a maximum concentration of 17,387 cells L⁻¹ at our nearshore site in Monterey Bay. Peak cell densities occurred in the month of July and were associated with elevated shellfish toxicity in the summers of 2004 and 2005. When A. catenella was present, particulate PSTs were detected 71% of the time and reached a maximum concentration of 962 ng STXeq L⁻¹. Of the 13 species tested, we frequently detected PSTs in Pacific sardines (Sardinops sagax; maximum 250 μg STXeq 100 g⁻¹), northern anchovies (Engraulis mordax; maximum 23.2 μg STXeq 100 g⁻¹), brown rock crabs (Cancer antennarius; maximum 49.3 μg STXeq 100 g⁻¹) and red rock crabs (C. productus; 23.8 μg STXeq 100 g⁻¹). PSTs were also present in one sample of Pacific herring (Clupea pallas; 13.3 μg STXeq 100 g⁻¹) and one sample of English sole (Pleuronectes vetulus; 4.5 μg STXeq 100 g⁻¹), and not detected in seven other species of flatfish tested. The presence of PSTs in several of these organisms reveals that toxins produced by A. catenella are more prevalent in California food webs than previously thought and also indicates potential routes of toxin transfer to higher trophic levels. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Vision in the lanternfish Stenobrachius leucopsarus (Myctophidae)

The visual system of the midwater fish Stenobrachius leucopsarus (Myctophidae) has been studied by biochemical, anatomical, and electrophysiological methods. Partial bleaching analysis in the presence of hydroxylamine showed that the eye contains a single extractable photopigment, based on retinal and absorbing maximally at 492 nm. The photoreceptor population consists entirely of rods, approximately 5.0×10⁵ rods per mm² and 1.8×10⁷ per retina. Visual sensitivity is enhanced by the lack of pigment granules in the pigment epithelium and the presence of a choroidal tapetum lucidum. Influenced by the high concentration of visual pigment, the spectral sensitivity has a broad plateau between 460 and 540 nm, which suggests that the eye retains high sensitivity to a diversity of bioluminescent stimuli and depth-attenuated solar light.     

Intracellular cyanobiont Richelia intracellularis: ultrastructure and immuno-localisation of phycoerythrin,nitrogenase, Rubisco and glutamine synthetase

In marine tropical or subtropical plankton the filamentous, heterocyst-forming cyanobacterium Richelia intracellularis forms a symbiosis with the diatom Rhizosolenia clevei. An ultrastructural analysis of the apex of Rhizosolenia clevei showed that the cytoplasm in that particular part of the cell was present only where the cyanobiont was located. The cyanobiont was, however, always outside the host cytoplasm. Vegetative cells as well as the heterocysts of the cyanobiont were devoid of gas vesicles and cyanophycin granules, while carboxysomes and large glycogen granules were common. The cyanobacterial cell wall apparently remained intact in both vegetative and heterocyst cells. In green excitation light the heterocysts and vegetative cells emitted a bright yellow fluorescence, indicating that both cell types possessed high concentrations of the pigment phycoerythrin (PE) commonly associated with photosystem (PS) II. The presence of this pigment in both cell types was verified by immunogold localisation. Using the same technique, the nitrogenase (dinitrogenase reductase) enzyme was shown to be exclusively present in the heterocysts, while Rubisco was localised primarily to the carboxy-somes, which were only detected in vegetative cells. Using an antiserum against the ammonia assimilating enzyme glutamine synthetase (GS), we could demonstrate very low levels of this enzyme, indicating repression of GS in the cyanobiont.     

The osphradium in Placopecten magellanicus and Pecten maximus (Bivalvia,Pectinidae): histology,ultrastructure, and implications for spawning synchronisation

The bivalve osphradium is a band of putatively sensory tissue located in the gill axis, whose function is uncertain. In the present study, extending from 1987 to 1994, anatomical, histological, and electron microscopical techniques were used to elucidate the structure and ultrastructure of the osphradium in hatchery Pecten maximus L. and Placopecten magellanicus (Gmelin) (collected from Passamaquoddy Bay, New Brunswick, Canada). The osphradium consists of two distinct regions which run longitudinally on both sides of each gill axis: the osphradial ridge, and the dorsal tuft cilia region. The osphradial ridge was largely devoid of cilia other than those of the few free nerve fibres. The dorsal tuft cilia region contained free nerve fibres and ciliary tufts, separated by undifferentiated epithelial cells. No paddle cilia were observed under isosmotic fixation conditions, although under hypotonic conditions such cilia were quite common, suggesting an artefactual nature. Most of the cells of the osphradial ridge were highly secretory, the principal products being large pigment granules (in Pecten maximus) directly secreted by the Golgi bodies, and numerous small, electron-dense vesicles. These vesicles were arranged along extensive microtubule arrays in the basal region, indicative of axonal transport. These data support and extend Haszprunar's hypothesis of the role of the osphradium in the reception of chemical spawning cues and in the synchronization of gamete emission. Together with independent data on nerve pathways, osphradial sensory modalities, and monoamine localisation, an anatomical pathway and neurophysiological mediator are postulated.     

Field depuration and biotransformation of paralytic shellfish toxins in scallop<Emphasis Type="Italic"> Chlamys nobilis</Emphasis> and green-lipped mussel<Emphasis Type="Italic"> Perna viridis</Emphasis>

Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration was characterized by a rapid loss within the first day, followed by a secondary slower loss of toxins. In the fast depuration phase, scallops detoxified PSTs more quickly than the mussels (depuration rate constants for scallops and mussels were 1.16 day^–1 and 0.87 day^–1, respectively). In contrast, the mussels detoxified PSTs more quickly than the scallops in the slow depuration phase, and the calculated depuration rate constants (mean+SE) from day 2 to day 13 were 0.063+0.009 day^–1 and 0.040+0.019 day^–1 for mussels and scallops, respectively. The differences in the appearances of gonyautoxins, GTX2 and GTX3, and their decarbamoyl derivatives, dcGTX2, dcGTX3 and GTX5, which are all derivatives of C2 toxin, indicated active and species-specific biotransformation of the algal toxins in the two bivalves. In both species of bivalves, the non-viscera tissue contained fewer toxins and lower concentrations than the viscera-containing tissue compartment. In scallops, very little toxin was distributed in the adductor muscle. In mussels, most of the PSTs were found in the digestive gland with significant transport of toxins into the digestive gland from other tissues during the course of depuration. The toxin profiles of scallops and mussels differed from each other and from that of the toxic algae fed. A significant fraction of GTX5 was detected in the mussels but not in the scallops. Our study demonstrates a species specificity in the depuration kinetics, biotransformation and tissue distribution of PSTs among different bivalves.Communicated by T. Ikeda, Hakodate     

Role of secondary metabolites and pigments in the epidermal tissues, ripe ovaries, viscera, gut contents and diet of the sea cucumber Holothuria atra

The epidermal tissues, ovaries, viscera, gut contents, and the diet of Holothuria atra (Jaeger), collected from the Great Barrier Reef (GBR), Australia, just prior to spawning, contained carotenoid pigments, mycosporine-like amino acids (MAAs) and three other metabolites in varying proportions. Mycosporine-glycine (mycosporine-gly), palythine, asterina-330, shinorine, mycosporine-2-gly, porphyra-334, and palythinol were present in the epidermal tissues. Mycosporine-gly was the major MAA in the epidermal tissues, gut contents and the diet, and the only MAA present in the ripe ovaries and the viscera. Gadusol, a metabolite with antioxidant properties, which co-occurs with MAAs, and in certain instances with trace amounts of 6-deoxygadusol in unfertilised and fertilised eggs and developing larvae of some marine invertebrates and vertebrates, was absent from H. atra. However, 6-deoxygadusol, with similar physical and chemical properties to gadusol, and the proposed biosynthetic precursor of MAAs, was present, but only in the ovaries. This is first report of the presence of this proposed biogenetic precursor of MAAs in significant amounts in a marine invertebrate. Approximately 90% of the total carotenoids of the epidermal tissues, ovaries and viscera of H. atra are highly oxidised, the main component being astaxanthin followed by canthaxanthin. These were the major carotenoids present in the ovaries and the total carotenoid content was highest in the ovaries. β-carotene, a common egg carotenoid, was present in trace quantities in the ovaries but, along with the xanthophylls lutein and zeaxanthin, it occurred in significant amounts in the gut contents and the diet of H. atra. Carotenoid patterns in the epidermal tissues and viscera were strikingly similar, containing β-carotene, canthaxanthin, astaxanthin, zeaxanthin, lutein, phoenicoxanthin and echinenone. The alkaloids homarine and isomeric trigonelline, (structurally unrelated to MAAs) and 6-deoxygadusol were present only in the ovaries and the viscera. It is suggested that these metabolites and pigments are either involved with photoprotection or reproduction, or associated with both processes. The origin, biogenetic relationships and the roles of these metabolites and pigments in H. atra are discussed. Received: 21 November 1997 / Accepted: 14 September 1998