A lectin from Crenomytilus grayanus as a probe for the detection of widespread cancerous and metastatic cells.

A novel Gal-binding lectin from mussels (Crenomytilus grayanus, CGL) with 6 binding sites in the dimeric structure has been previously shown to have antifungal, anticancer, and antibacterial activities. In this study, a glycan array was used to confirm that CGL recognizes a range of non-reducing end α- or ß-linked Gal glycans on normal cells but not sialic acid-capped glycans. This finding suggests that CGL has potential in the tumor detection due to the hyper-sialylation present in cell surface glycans from cancer cells. To evaluate the feasibility of this possibility, we labeled CGL with biotin and then mixed it with streptavidin-horseradish peroxidase (HRP) to create a CGL-biotin-SP complex as a probe for use in enzyme-linked lectin assays. CGL-biotin-SP successfully distinguished not only HeLa cells and de-sialylated HeLa cells that mimic normal cell surface glycans but also lung and breast cancer cells with different metastatic abilities. This work provides the insights into a new Gal-binding lectin by establishing its specificity and also demonstrates practical applications in cancer diagnosis greater than other reported lectins.

Insights into symbiotic interactions from metatranscriptome analysis of deep-sea mussel Gigantidas platifrons under long-term laboratory maintenance.

Symbioses between invertebrates and chemosynthetic bacteria are of fundamental importance in deep-sea ecosystems, but the mechanisms that enable their symbiont associations are still largely undescribed, owing to the culturable difficulties of deep-sea lives. Bathymodiolinae mussels are remarkable in their ability to overcome decompression and can be maintained successfully for an extended period under atmospheric pressure, thus providing a model for investigating the molecular basis of symbiotic interactions. Herein, we conducted metatranscriptome sequencing and gene co-expression network analysis of Gigantidas platifrons under laboratory maintenance with gradual loss of symbionts. The results revealed that one-day short-term maintenance triggered global transcriptional perturbation in symbionts, but little gene expression changes in mussel hosts, which were mainly involved in responses to environmental changes. Long-term maintenance with depleted symbionts induced a metabolic shift in the mussel host. The most notable changes were the suppression of sterol biosynthesis and the complementary activation of terpenoid backbone synthesis in response to the reduction of bacteria-derived terpenoid sources. In addition, we detected the upregulation of host proteasomes responsible for amino acid deprivation caused by symbiont depletion. Additionally, a significant correlation between host microtubule motor activity and symbiont abundance was revealed, suggesting the possible function of microtubule-based intracellular trafficking in the nutritional interaction of symbiosis. Overall, by analyzing the dynamic transcriptomic changes during the loss of symbionts, our study highlights the nutritional importance of symbionts in supplementing terpenoid compounds and essential amino acids and provides insight into the molecular mechanisms and strategies underlying the symbiotic interactions in deep-sea ecosystems.

Aromatic Polyketides from the Deep-Sea Cold-Seep Mussel Associated Endozoic Fungus Talaromyces minioluteus CS-138.

Five new aromatic polyketides, including a unique benzofuran derivative, talarominine A (1), and four chromone analogs talamins A-D (2-5), along with one known related metabolite, 5-hydroxy-7-methoxy-2,3-dimethylchromone (6), were isolated and identified from the Talaromyces minioluteus CS-138, an endozoic fungus obtained from the deep-sea cold seep mussel Gigantidas platifrons. Their chemical structures were elucidated by detailed analysis of their NMR spectra, HRESIMS and X-ray crystallographic data, and by comparison with literature data as well. The antibacterial and DPPH scavenging activities of compounds 1-6 were evaluated. Compounds 1-3 showed inhibitory activity against some of the tested bacteria whereas compounds 2 and 5 showed potent DPPH radical scavenging activities, which were better than that of the positive control butylated hydroxytoluene (BHT). This work is likely the first report on marine natural products of mussel-derived fungus living in cold seep environments.

Bioaccumulation and retention kinetics of trace elements in the horse mussels Modiolus micropterus exposed to different environmental conditions.

Bioaccumulation studies of Zn and 137Cs by the horse mussel (Modiolus micropterus) were conducted in a laboratory that used radiotracer. The study has been carried out on the effect of cesium and zinc concentrations and the effect of sea seawater salinity on the ability of M. micropterus to accumulate these two contaminants. The uptake of Zn and Cs according to the one-compartment model and the experiment was carried out until the steady-state conditions were reached. The concentration factor at steady-state Zn is 31.94-45.54 mL. g-1 and 23.22-33.26 mL. g-1 which are influenced by the concentration and salinity of seawater, respectively. The concentration factor of 137Cs at steady-state conditions due to changes in concentration and salinity is 3.34-7.55 mL. g-1 and 4.23-9.66 mL. g-1, respectively. The release rates of Zn were 30-47 % and 39-49 % at various concentrations and salinity. The depuration rate from concentration reaching 60 % and salinity at ranges 43-52 % was observed within 10 days after exposure. On the other hand, the release rates of 137Cs were 60 % and 43-52 % at various changes in the concentration and salinity of seawater.

Fatty acid response of the invasive bivalve Limnoperna fortunei fed with Microcystis aeruginosa exposed to high temperature.

The success of Limnoperna fortunei as an invasive freshwater bivalve species is related to its physiological plasticity to endure changes in environmental conditions. The aim of this study was to investigate the physiological responses of L. fortunei after feeding on Microcystis aeruginosa grown at 26 °C (control) and 29 °C during 10 days. At the beginning, we measured biomass, fatty acids (FAs) composition on Cyanobacteria grown at both temperatures at different time intervals. Afterwards, mussels were fed with the thawed M. aeruginosa cells and their FA profile was measured after 15 days of feeding. M. aeruginosa exposed to 29 °C had the highest content of the FAs 18:2ω6 and cis-18:1ω9. The FA profile of the consumer L. fortunei fed with M. aeruginosa cultures grown at 29 °C was also significantly different to those fed with cultures grown at 26 °C, with a significant increased Eicosapentaenoic acid (EPA, 20:5ω3) and Arachidonic acid (ARA, 20:4ω6) concentrations. L. fortunei was already known to be physiologically adapted to live at 29 °C, but our results also shown a high biosynthesis of EPA and ARA (increase of 70 and 40% respectively, compared with 26 °C) and avoided the lipid peroxidation of both FAs. This increased EPA and ARA biosynthesis may be an important source of ω3 and ω6 polyunsaturated FAs (PUFAs) for higher trophic levels, such as the pelagic fishes or birds that mainly prey on these mussels. The transfer of the cyanobacterial response at higher temperature to higher trophic levels will influence the overall functioning of freshwater bodies.

Modeling of the mussel catch muscle contractile apparatus in actomyosin suspension.

In this paper, we tried to create a contractile model from proteins of the catch muscle of the Gray mussel, similar to the well-described suspension contractile model of vertebrate skeletal muscles. This model makes it possible to characterize the processes in the reconstructed contractile apparatus with the help of monitoring the two characteristics of muscle suspensions - the optical density and the particle size. Contractile model of the catch muscle we constructed was the simplest model consisting of two proteins, actin and myosin. During this work we compared the optical manifestations of the contraction and relaxation states of constructed model with earlier data on the actomyosin suspension of skeletal muscles. It appeared that the approach used in the study of skeletal muscle actomyosin relaxing - the use of an increased amount of ATP - cannot be applied to the contractile model of the molluscan catch muscle. Nevertheless we managed to reach relaxed state of this model with modifying calcium concentration. As a result, we laid the foundation for further reconstruction of the third state of the catch muscle - the catch tone.

Invasive species versus pollutants: Potential of Limnoperna fortunei to degrade glyphosate-based commercial formulations.

The intensive use of glyphosate in industrial agriculture may lead to freshwater contamination, encouraging studies of its toxic effect on non-target aquatic organisms. Glyphosate-based commercial formulations contain adjuvants, making them even more toxic than the active ingredient (a.i.) itself. The golden mussel Limnoperna fortunei is a freshwater invasive species which has been found to increase glyphosate dissipation in water and to accelerate eutrophication. The aim of this study is to evaluate the capability of L. fortunei to reduce the concentration of glyphosate in two commercial formulations, Roundup Max® and Glifosato Atanor®. Results were compared with the decay of the a.i. alone and in presence of mussels. Evasive response and toxicity tests were performed in a first set of trials to analyze the response of L. fortunei exposed to Roundup Max® and Glifosato Atanor®. Subsequently, we conducted a 21-day degradation experiment in 2.6-L microcosms applying the following treatments: 6 mg L-1 of technical-grade glyphosate (G), Glifosato Atanor® (A), Roundup Max® (R), 20 mussels in dechlorinated tap water (M), and the combination of mussels and herbicide either in the technical-grade (MG) or formulated form (MA and MR) (all by triplicate). Samples were collected at days 0, 1, 7, 14 and 21. No significant differences in glyphosate decay were found between treatments with mussels (MG: 2.03 ± 0.40 mg L-1; MA: 1.60 ± 0.32 mg L-1; MR: 1.81 ± 0.21 mg L-1), between glyphosate as a.i. and the commercial formulations, and between the commercial formulations, suggesting that the adjuvants did not affect the degrading potential of L. fortunei. In addition to the acceleration of glyphosate dissipation in water, there was an increase in the concentration of dissolved nutrients in water (N-NH4+ and P-PO43-) even higher than that caused by the filtering activity of the mussels, probably resulting from stress or from the degradation of glyphosate and adjuvants. We believe that a larger bioavailability of these nutrients due to glyphosate metabolization mediated by mussels would accelerate eutrophication processes in natural water bodies. The approach used here, where L. fortunei was exposed to two commercial formulations actually used in agricultural practices, sheds light on the potential impact of glyphosate decay on water bodies invaded by this species.

Molecular Identification, Characterization, and Expression Analysis of a Metallothionein Gene from Septifer virgatus.

This study provides a preliminary characterization of a metallothionein (MT) gene in Septifer virgatus and highlights its potential use in biomonitoring. The full-length SvMT cDNA and the complete sequence of the SvMT gene were identified using reverse transcriptase PCR coupled with the rapid amplification of cDNA ends and the primer walking method. The SvMT cDNA encodes a protein of 72 amino acids having nine classical Cys-X-Cys motifs. Moreover, the deduced amino acids contained the conserved motif (Cys-x-Cys-x(3)-Cys-Thr-Gly-x(3)-Cys-x-Cys-x(3)-Cys-x-Cys-Lys) of MT family 2. Its molecular mass and isoelectric point were estimated to be 7.01 kDa and 7.00, respectively. BLAST-based searching indicated that SvMT shared 81.0% amino acid sequence identity with Mytilus edulis MT-20-II. The SvMT gene has three coding exons and two introns. After exposure to 1 mg/L cadmium chloride, the expression of SvMT increased 15-fold by 3 days (d), with a maximum expression of 27-fold by 5 d compared with the pre-exposure level. After exposure to 2 mg/L zinc chloride, the expression of SvMT increased 2.5-fold by 3 d and 4.7-fold by 5 d compared with the pre-exposure level. A significant increase in the expression level of SvMT mRNA was observed after the exposure of S. virgatus to the combination of 0.003 mg/L cadmium chloride and 0.2 mg/L zinc chloride compared with the pre-exposure level. Our work indicates that the SvMT gene is associated with stress responses and could be a potential biomarker for marine pollution.

Comparative proteomics of related symbiotic mussel species reveals high variability of host-symbiont interactions.

Deep-sea Bathymodiolus mussels and their chemoautotrophic symbionts are well-studied representatives of mutualistic host-microbe associations. However, how host-symbiont interactions vary on the molecular level between related host and symbiont species remains unclear. Therefore, we compared the host and symbiont metaproteomes of Pacific B. thermophilus, hosting a thiotrophic symbiont, and Atlantic B. azoricus, containing two symbionts, a thiotroph and a methanotroph. We identified common strategies of metabolic support between hosts and symbionts, such as the oxidation of sulfide by the host, which provides a thiosulfate reservoir for the thiotrophic symbionts, and a cycling mechanism that could supply the host with symbiont-derived amino acids. However, expression levels of these processes differed substantially between both symbioses. Backed up by genomic comparisons, our results furthermore revealed an exceptionally large repertoire of attachment-related proteins in the B. thermophilus symbiont. These findings imply that host-microbe interactions can be quite variable, even between closely related systems.

A GM1b/asialo-GM1 oligosaccharide-binding R-type lectin from purplish bifurcate mussels Mytilisepta virgata and its effect on MAP kinases.

A 15-kDa lectin, termed SeviL, was isolated from Mytilisepta virgata (purplish bifurcate mussel). SeviL forms a noncovalent dimer that binds strongly to ganglio-series GM1b oligosaccharide (Neu5Acɑ2-3Galß1-3GalNAcß1-4Galß1-4Glc) and its precursor, asialo-GM1 (Galß1-3GalNAcß1-4Galß1-4Glc). SeviL also interacts weakly with the glycan moiety of SSEA-4 hexaose (Neu5Acα2-3Galß1-3GalNAcß1-3Galα1-4Galß1-4Glc). A partial protein sequence of the lectin was determined by mass spectrometry, and the complete sequence was identified from transcriptomic analysis. SeviL, consisting of 129 amino acids, was classified as an R(icin B)-type lectin, based on the presence of the QxW motif characteristic of this fold. SeviL mRNA is highly expressed in gills and, in particular, mantle rim tissues. Orthologue sequences were identified in other species of the family Mytilidae, including Mytilus galloprovincialis, from which lectin MytiLec-1 was isolated and characterized in our previous studies. Thus, mytilid species contain lectins belonging to at least two distinct families (R-type lectins and mytilectins) that have a common ß-trefoil fold structure but differing glycan-binding specificities. SeviL displayed notable cytotoxic (apoptotic) effects against various cultured cell lines (human breast, ovarian, and colonic cancer; dog kidney) that possess asialo-GM1 oligosaccharide at the cell surface. This cytotoxic effect was inhibited by the presence of anti-asialo-GM1 oligosaccharide antibodies. With HeLa ovarian cancer cells, SeviL showed dose- and time-dependent activation of kinase MKK3/6, p38 MAPK, and caspase-3/9. The transduction pathways activated by SeviL via the glycosphingolipid oligosaccharide were triggered apoptosis. DATABASE: Nucleotide sequence data have been deposited in the GenBank database under accession numbers MK434191, MK434192, MK434193, MK434194, MK434195, MK434196, MK434197, MK434198, MK434199, MK434200, and MK434201.

Functional diversity enables multiple symbiont strains to coexist in deep-sea mussels.

Genetic diversity of closely related free-living microorganisms is widespread and underpins ecosystem functioning, but most evolutionary theories predict that it destabilizes intimate mutualisms. Accordingly, strain diversity is assumed to be highly restricted in intracellular bacteria associated with animals. Here, we sequenced metagenomes and metatranscriptomes of 18 Bathymodiolus mussel individuals from four species, covering their known distribution range at deep-sea hydrothermal vents in the Atlantic. We show that as many as 16 strains of intracellular, sulfur-oxidizing symbionts coexist in individual Bathymodiolus mussels. Co-occurring symbiont strains differed extensively in key functions, such as the use of energy and nutrient sources, electron acceptors and viral defence mechanisms. Most strain-specific genes were expressed, highlighting their potential to affect fitness. We show that fine-scale diversity is pervasive in Bathymodiolus sulfur-oxidizing symbionts, and hypothesize that it may be widespread in low-cost symbioses where the environment, rather than the host, feeds the symbionts.

Protein composition of thick filaments from molluscan catch muscle and the role of twitchin in the catch-state formation.

In the work, we performed densitometry of thick filaments of the Gray's mussel catch muscle; densitometry included determination of electrophoretic dye binding constants of proteins. The results of densitometry showed that the content of twitchin in thick filaments is significantly (10 times) lower than the content of myosin. We performed an in vitro simulation of the contractile apparatus of the catch muscle and showed that with such content, links formed by twitchin cannot stop "relaxation". So, we doubt that the role of twitchin in the formation of the catch state is to form load-bearing links between thin and thick filaments that keep the muscle in the contracted state. At the same time, densitometry has shown that the content of the unique catch-muscle protein - myorod - significantly exceeds the content of twitchin and reaches the level of myosin. Like twitchin, myorod is capable of forming regulated cross-links between thick and thin filaments. Such a high content of this protein may indicate that it is myorod, and not twitchin, that is responsible for the formation of catch load-bearing cross-links.

Protein expression profiles in Bathymodiolus azoricus exposed to cadmium.

Proteomic changes in the "gill-bacteria complex" of the hydrothermal vent mussel B. azoricus exposed to cadmium in pressurized chambers ((Incubateurs Pressurises pour l'Observation en Culture d'Animaux Marins Profonds - IPOCAMP) were analyzed and compared with the non-exposed control group. 2-D Fluorescence Difference Gel Electrophoresis (2D-DIGE) showed that less than 1.5% of the proteome of mussels and symbiotic bacteria were affected by a short-term (24 h) Cd exposure. Twelve proteins of the more abundant differentially expressed proteins of which six were up-regulated and six were down-regulated were excised, digested and identified by mass spectrometry. The identified proteins included structural proteins (actin/actin like proteins), metabolic proteins (calreticulin/calnexin, peptidyl-prolyl cis-trans isomerase, aminotransferase class-III, electron transfer flavoprotein, proteasome, alpha-subunit and carbonic anhydrase) and stress response proteins (chaperone protein htpG, selenium-binding protein and glutathione transferases). All differently expressed proteins are tightly connected to Cd exposure and are affected by oxidative stress. It was also demonstrated that B. azoricus was well adapted to Cd contamination therefore B. azoricus from hydrothermal vent areas may be considered a good bioindicator.

Gill symbionts of the cold-seep mussel Bathymodiolus platifrons: Composition, environmental dependency and immune control.

Deep-sea Bathymodiolus mussels depend on the organic carbon supplied by symbionts inside their gills. In this study, optimized methods of quantitative real-time PCR and fluorescence in situ hybridization targeted to both mRNA and 16S rRNA were used to investigate the gill symbionts of the cold-seep mussel Bathymodiolus platifrons, including species composition, environmental dependency and immune control by the host. Our results showed that methanotrophs were the major symbiotic bacteria in the gills of B. platifrons, while thiotrophs were scarce. In the mussels freshly collected from the deep sea, methanotrophs were housed in bacteriocytes in a unique circular pattern, and a lysosome-related gene (VAMP) encoding a vesicle-associated membrane protein was expressed at a high level and presented exactly where the methanotrophs occurred. After the mussels were reared for three months in aquaria without methane supply, the abundance of methanotrophs decreased significantly and their circle-shaped distribution pattern disappeared; in addition, the expression of VAMP decreased significantly. These results suggest that the symbiosis between B. platifrons and methanotrophs is influenced by the environment and that the lysosomal system plays an important immune role in controlling the abundance of endosymbionts in host. This study provides a reliable method for investigating symbionts in deep-sea mussels and enriches the knowledge about symbionts in B. platifrons.

Functional characterization of the GABA transporter GAT-1 from the deep-sea mussel Bathymodiolus septemdierum.

Mammalian γ-aminobutyric acid (GABA) transporter subtype 1 (GAT-1) is a specific transporter for GABA, an inhibitory neurotransmitter in GABA-ergic neurons. GAT-1 belongs to the GAT group, in which five related transporters, GAT-2, GAT-3, GAT-4, CT1, and TAUT are known in mammals. By contrast, the deep-sea mussel, Bathymodiolus septemdierum has only two GAT group members, BsGAT-1 and BsTAUT, and their function in environmental adaptation is of interest to better understand the physiology of deep-sea organisms. Compared with BsTAUT, the function of BsGAT-1 is unknown. Here, we report the functional characterization of BsGAT-1. Analyses of BsGAT-1 expressed in Xenopus oocytes showed that it could transport GABA in a Na+- and Cl--dependent manner, with Km and Vmax values of 0.58 µM and 1.97 pmol/oocyte/h, respectively. BsGAT-1 activity was blocked by the GAT-1 selective inhibitors SKF89976A and ACHC. Competition assays indicated that BsGAT-1 has no affinity for taurine and thiotaurine. These characteristics were common with those of mammalian GAT-1, suggesting its conserved function in the nervous system. However, BsGAT-1 showed a certain affinity for hypotaurine, which is involved in sulfide detoxification in hydrothermal vent-specific animals. This result suggests an additional role for BsGAT-1 in sulfide detoxification, which may be specific to the deep-sea mussel. In a tissue distribution analysis, BsGAT-1 mRNA expression was observed in various tissues. The expression in the adductor and byssus retractor muscles, labial palp, and foot, which possibly contain ganglia, suggested a function in the neural system, while BsGAT-1 expression in other tissues might be related to sulfide detoxification.

Activity Dependence of a Novel Lectin Family on Structure and Carbohydrate-Binding Properties.

A GalNAc/Gal-specific lectins named CGL and MTL were isolated and characterized from the edible mussels Crenomytilus grayanus and Mytilus trossulus. Amino acid sequence analysis of these lectins showed that they, together with another lectin MytiLec-1, formed a novel lectin family, adopting ß-trefoil fold. In this mini review we discuss the structure, oligomerization, and carbohydrate-binding properties of a novel lectin family. We describe also the antibacterial, antifungal, and antiproliferative activities of these lectins and report about dependence of activities on molecular properties. Summarizing, CGL, MTL, and MytiLec-1 could be involved in the immunity in mollusks and may become a basis for the elaboration of new diagnostic tools or treatments for a variety of cancers.

Mutagenesis Studies and Structure-function Relationships for GalNAc/Gal-Specific Lectin from the Sea Mussel Crenomytilus grayanus.

The GalNAc/Gal-specific lectin from the sea mussel Crenomytilus grayanus (CGL) with anticancer activity represents а novel lectin family with ß-trefoil fold. Earlier, the crystal structures of CGL complexes with globotriose, galactose and galactosamine, and mutagenesis studies have revealed that the lectin contained three carbohydrate-binding sites. The ability of CGL to recognize globotriose (Gb3) on the surface of breast cancer cells and bind mucin-type glycoproteins, which are often associated with oncogenic transformation, makes this compound to be perspective as a biosensor for cancer diagnostics. In this study, we describe results on in silico analysis of binding mechanisms of CGL to ligands (galactose, globotriose and mucin) and evaluate the individual contribution of the amino acid residues from carbohydrate-binding sites to CGL activity by site-directed mutagenesis. The alanine substitutions of His37, His129, Glu75, Asp127, His85, Asn27 and Asn119 affect the CGL mucin-binding activity, indicating their importance in the manifestation of lectin activity. It has been found that CGL affinity to ligands depends on their structure, which is determined by the number of hydrogen bonds in the CGL-ligand complexes. The obtained results should be helpful for understanding molecular machinery of CGL functioning and designing a synthetic analog of CGL with enhanced carbohydrate-binding properties.

Biochemical responses of the golden mussel Limnoperna fortunei under dietary glyphosate exposure.

The aim of this study was to analyze the biochemical alterations in the golden mussel Limnoperna fortunei under dietary glyphosate exposure. Mussels were fed during 4 weeks with the green algae Scenedesmus vacuolatus previously exposed to a commercial formulation of glyphosate (6 mg L-1 active principle) with the addition of alkyl aryl polyglycol ether surfactant. After 1, 7, 14, 21 and 28 days of dietary exposure, glutathione-S-transferase (GST), catalase (CAT), superoxide dismutase (SOD), acetylcholinesterase (AChE), carboxylesterases (CES) and alkaline phosphatase (ALP) activities, glutathione (GSH) content and damage to lipids and proteins levels were analyzed. A significant increase (72%) in the GST activity and a significant decrease (26%) in the CES activity in the mussels fed on glyphosate exposed algae for 28 days were observed. The ALP activity was significantly increased at 21 and 28 days of dietary exposure (48% and 72%, respectively). GSH content and CAT, SOD and AchE activities did not show any differences between the exposed and non exposed bivalves. No oxidative damage to lipids and proteins, measured as TBARS and carbonyl content respectively, was observed in response to glyphosate dietary exposure. The decrease in the CES activity and the increases in GST and ALP activities observed in L. fortunei indicate that dietary exposure to glyphosate provokes metabolic alterations, related with detoxification mechanisms.

Trace metals and oxidative status in soft tissues of caged mussels (Aulacomya atra) on the North Patagonian coastline.

This study investigated metal accumulation and oxidative effects in mantle, gill and digestive gland of the ribbed mussel Aulacomya atra from the Argentinean North Patagonian coastline. Mussels were transplanted over an 18-month period from a site with low anthropogenic impact to a harbor site with higher seawater concentration of aluminum, chromium, copper, manganese, nickel and zinc. Total trace metal concentration in seawater did not change throughout the 18-month transplant in either site. A. atra bioaccumulated metals in digestive gland, gills and mantle at different levels. Digestive gland had the highest concentration of metals, especially towards the end of the transplant experiment in the harbor area. Mussels transplanted to the harbor site experienced an upregulation in their antioxidant system, which likely explains the lack of oxidative damage to lipids despite higher metal accumulation. These results demonstrate that A. atra selectively accumulates metals from the water column and their prooxidant effects depend on the tissue antioxidant defenses and the exposure time.

Accumulation and Tissue Distribution of Dinophysitoxin-1 and Dinophysitoxin-3 in the Mussel Crenomytilus grayanus Feeding on the Benthic Dinoflagellate Prorocentrum foraminosum.

A DTX-1-producing microalga, Prorocentrum foraminosum, from Peter the Great Bay, Sea of Japan, was fed to Gray's mussels, Crenomytilus grayanus, for 12 days. An increase in DTX-1 and 7-O-acyl-DTX-1 (DTX-3) was observed in the digestive gland, kidneys, and gills. The digestive gland accumulated 91-100% of DTX-1 + DTX-3; and kidneys and gills accumulated, up to 8.5% and 4.3%, respectively. The kidneys had a distinctive pattern of toxin accumulation where the concentration of DTX-1 did not grow significantly after the eighth day of feeding, indicating the potential of DTX-1 elimination. The digestive gland and gills predominantly accumulated DTX-1, with a dramatic increase between Days 8 and 12. The DTX-3 content was highest in the digestive gland. The composition of DTX-3 in the acyl groups was similar for the digestive gland and kidneys, and did not change during feeding. The total toxin uptake of mussels exceeded the total toxin content from ingested cells by 2.4 times, showing that toxins may have accumulated from the seawater. This assumption needs to be further proved. The muscle, gonads, and mantle remained free of toxins. No genotoxic effect was observed in the gills and digestive gland.