Bivalve Crassostrea Gigas Research Articles

Mitochondrial responses to constant and cyclic hypoxia depend on the oxidized fuel in a hypoxia-tolerant marine bivalve Crassostrea gigas

Sessile benthic organisms like oysters inhabit the intertidal zone, subject to alternating hypoxia and reoxygenation (H/R) episodes during tidal movements, impacting respiratory chain activities and metabolome compositions. We investigated the effects of constant severe hypoxia (90 min at ~ 0% O2 ) followed by 10 min reoxygenation, and cyclic hypoxia (5 cycles of 15 min at ~ 0% O2 and 10 min reoxygenation) on isolated mitochondria from the gill and the digestive gland of Crassostrea gigas respiring on pyruvate, palmitate, or succinate. Constant hypoxia suppressed oxidative phosphorylation (OXPHOS), particularly during Complex I-linked substrates oxidation. It had no effect on mitochondrial reactive oxygen species (ROS) efflux but increased fractional electron leak (FEL). In mitochondria oxidizing Complex I substrates, exposure to cyclic hypoxia prompted a significant drop after the first H/R cycle. In contrast, succinate-driven respiration only showed significant decline after the third to fifth H/R cycle. ROS efflux saw little change during cyclic hypoxia regardless of the oxidized substrate, but Complex I-driven FEL tended to increase with each subsequent H/R cycle. These observations suggest that succinate may serve as a beneficial stress fuel under H/R conditions, aiding in the post-hypoxic recovery of oysters by reducing oxidative stress and facilitating rapid ATP re-synthesis. The impacts of constant and cyclic hypoxia of similar duration on mitochondrial respiration and oxidative lesions in the proteins were comparable indicating that the mitochondrial damage is mostly determined by the lack of oxygen and mitochondrial depolarization. The ROS efflux in the mitochondria of oysters was minimally affected by oxygen fluctuations indicating that tight regulation of ROS production may contribute to robust mitochondrial phenotype of oysters and protect against H/R induced stress.

Larvae of a marine gastropod and a marine bivalve share common gene expression signatures during metamorphic competence

Many aquatic invertebrates undergo an indirect development, a biphasic life cycle which encompasses the transformation of free-swimming larvae into benthic juveniles via settlement and metamorphosis. During this transition, metamorphic competence is a crucial developmental stage that allows larvae to swim and feed in the planktonic realm while retaining the ability to settle and metamorphose in response to environmental cues. Although there have been substantial efforts to decipher the molecular mechanisms underlying this event in several molluscan species, the conserved biological pathways that are crucial to enable this transition across species are not well understood. Here, we performed a comparative analysis of the developmental transcriptomes between bivalve Crassostrea gigas and gastropod Rapana venosa. We particularly explored the common gene expression signatures that may underlie their larval competence. We showed that, although the developmental transcriptomes differed remarkably between C. gigas and R. venosa, they likely shared a plethora of genes (n = 690) that exhibited similar expression signatures during their larval competence. Gene Ontology enrichment and expression analyses further indicated that competent larvae of both species exhibited up-regulation of pathways associated with response to stimuli, metal ion binding and transport, and neuronal development, but showed down-regulation of pathways that were mainly involved in cilium assembly and organ development. Using oyster and whelk as models, our study suggests that regulation of these conserved pathways is crucial for their subsequent settlement and metamorphosis and may represent a universal mechanism that enables the pelagic-to-benthic transition in a broader range of marine invertebrates.

Activity Levels of 210Po, 210Pb and Other Radionuclides (134Cs, 137Cs, 90Sr, 110mAg, 238U, 226Ra and 40K) in Marine Organisms From Coastal Waters Adjacent to Fuqing and Ningde Nuclear Power Plants (China) and Radiation Dose Assessment

With the rapid development of nuclear power, the radiation impacts on edible marine organisms, and the potential radiation risks to humans have become of considerable concern to public health. In this study, the activities of 210Po and 210Pb as well as those of other radionuclides in fishes (Mugil cephalus, Konosirus punctatus, Largehead hairtail, and Larimichthys polyactis), crustaceans (Mantis shrimp, Parapenaeopsis hardwickii, and Portunus trituberculatus), bivalves (Crassostrea gigas, Sinonovacula conzcta), and macroalgae (Gracilaria, Porphyra) collected in the coastal area adjacent to the Fuqing and Ningde nuclear power plants (NPPs) were determined. The activity range of 210Po and 210Pb was 0.60–48.09 and 0.07–2.76 Bq/kg freshweight, respectively, with 210Po/210Pb activity ratios of 1.1–189.7. The ranking of 210Po activity levels in marine organisms was bivalve mollusks > crustaceans > fishes > macroalgae. The calculated bioconcentration factors of 210Po and 210Pb were 636–44,944 and 3–1,226 L/kg, respectively. These values provide a new supplement to the IAEA reference database. The radiation dose rates for these marine organisms ranged from 0.037 to 1.531 μSv/h, which was much lower than the ERICA ecosystem screening benchmark of 10 μGy/h. The calculated committed effective dose received by humans from ingestion of these marine organisms was 0.06–2.99 mSv. Overall, 210Po was the dominant radiation dose contributor in marine organisms and humans, whereas the dose contributions from the artificial nuclides 90Sr and 137Cs were negligible.

First detection of OsHV-1 in the cephalopod Octopus vulgaris. Is the octopus a dead-end for OsHV-1?

The ostreid herpes virus (OsHV-1), associated with massive mortalities in the bivalve Crassostrea gigas, was detected for the first time in the cephalopod Octopus vulgaris. Wild adult animals from a natural breeding area in Spain showed an overall prevalence of detection of 87.5% between 2010 and 2015 suggesting an environmental source of viral material uptake. Overall positive PCR detections were significantly higher in adult animals (p = 0.031) compared to newly hatched paralarvae (62%). Prevalence in embryos reached 65%. Sequencing of positive amplicons revealed a match with the variant OsHV-1 µVar showing the genomic features that distinguish this variant in the ORF4. Gill tissues from adult animals were also processed for in situ hybridization and revealed positive labelling. Experimental exposure trials in octopus paralarvae were carried out by cohabitation with virus injected oysters and by immersion in viral suspension observing a significant decrease in paralarval survival in both experiments. An increase in the number of OsHV-1 positive animals was detected in dead paralarvae after cohabitation with virus injected oysters. No signs of viral replication were observed based on lack of viral gene expression or visualization of viral structures by transmission electron microscopy. The octopus response against OsHV-1 was evaluated by gene expression of previously reported transcripts involved in immune response in C. gigas suggesting that immune defences in octopus are also activated after exposure to OsHV-1.

Molecular Biomarkers of the Mitochondrial Quality Control Are Differently Affected by Hypoxia-Reoxygenation Stress in Marine Bivalves Crassostrea gigas and Mytilus edulis

Coastal environments commonly experience strong oxygen fluctuations. Resulting hypoxia/reoxygenation stress can negatively affect mitochondrial functions, since oxygen deficiency impairs ATP generation, whereas a surge of oxygen causes mitochondrial damage by oxidative stress. Marine intertidal bivalves are adapted to fluctuating oxygen conditions, yet the underlying molecular mechanisms that sustain mitochondrial integrity and function during oxygen fluctuations are not yet well understood. We used targeted mRNA expression analysis to determine the potential involvement of the mitochondrial quality control mechanisms in responses to short-term hypoxia (24 h at <0.01% O2) and subsequent reoxygenation (1.5 h at 21% O2) in two hypoxia-tolerant marine bivalves, the Pacific oysters Crassostrea gigas and the blue mussels Mytilus edulis. We hypothesized that the genes involved in the mitochondrial quality control will be upregulated during hypoxia, and the less hypoxia-tolerant of the two studied species (M. edulis) will show a stronger dependence on transcriptional upregulation of these pathways than C. gigas. To test these hypotheses, mRNA expression of 17 (C. gigas) and 11 (M. edulis) marker genes involved in mitochondrial fusion, fission, proteolysis and mitophagy was analyzed in the digestive gland of M. edulis and C. gigas in normoxia and during hypoxia-reoxygenation (H/R) stress. In the mussels, the mRNA expression of the transcripts related to mitochondrial dynamics and quality control was strongly altered during H/R stress showing a shift toward fission, suppression of fusion, an increase in mitochondrial proteolysis and onset of mitophagy. These changes indicate that H/R stress induces mitochondrial injury in M. edulis requiring upregulation of the protective mechanisms to segregate the dysfunctional mitochondria by fission and degrade the oxidative damaged proteins and/or organelles. Unlike mussels, the transcript levels of all studied genes in the oysters remained at the baseline (normoxic) levels during H/R stress. This muted transcriptional response of C. gigas is in agreement with earlier findings showing better ability to maintain cellular homeostasis and higher resistance to apoptosis during H/R stress in the oysters compared with the mussels. The revealed species-specific differences in the expression of the mitochondrial quality control pathways shed light on the potentially important mechanisms of mitochondrial protection against H/R-induced damage that might contribute to hypoxia tolerance in marine bivalves.

A-to-I editing of Malacoherpesviridae RNAs supports the antiviral role of ADAR1 in mollusks

BackgroundAdenosine deaminase enzymes of the ADAR family are conserved in metazoans. They convert adenine into inosine in dsRNAs and thus alter both structural properties and the coding potential of their substrates. Acting on exogenous dsRNAs, ADAR1 exerts a pro- or anti-viral role in vertebrates and Drosophila.ResultsWe traced 4 ADAR homologs in 14 lophotrochozoan genomes and we classified them into ADAD, ADAR1 or ADAR2, based on phylogenetic and structural analyses of the enzymatic domain. Using RNA-seq and quantitative real time PCR we demonstrated the upregulation of one ADAR1 homolog in the bivalve Crassostrea gigas and in the gastropod Haliotis diversicolor supertexta during Ostreid herpesvirus-1 or Haliotid herpesvirus-1 infection. Accordingly, we demonstrated an extensive ADAR-mediated editing of viral RNAs. Single nucleotide variation (SNV) profiles obtained by pairing RNA- and DNA-seq data from the viral infected individuals resulted to be mostly compatible with ADAR-mediated A-to-I editing (up to 97%). SNVs occurred at low frequency in genomic hotspots, denoted by the overlapping of viral genes encoded on opposite DNA strands. The SNV sites and their upstream neighbor nucleotide indicated the targeting of selected adenosines. The analysis of viral sequences suggested that, under the pressure of the ADAR editing, the two Malacoherpesviridae genomes have evolved to reduce the number of deamination targets.ConclusionsWe report, for the first time, evidence of an extensive editing of Malacoherpesviridae RNAs attributable to host ADAR1 enzymes. The analysis of base neighbor preferences, structural features and expression profiles of molluscan ADAR1 supports the conservation of the enzyme function among metazoans and further suggested that ADAR1 exerts an antiviral role in mollusks.

A Hybrid Phase I-Phase II Toxicity Identification Evaluation (TIE) for the Simultaneous Characterization and Identification of Toxicants of Concern in Coastal and Estuarine Environments.

A sequential TIE procedure combining in a single framework Phase I manipulations and Phase II methods, including chemical analyses and complementary Phase I treatments, was proposed for characterization and identification of toxicants of concern in estuarine environments. Interstitial water was chosen as test matrix and embryo-larval development with the bivalve Crassostrea gigas as toxicity endpoint. TIE treatments included addition of Ulva rigida and elution through zeolite column for addressing effects due to ammonia, addition of EDTA and elution through a Cation-Exchange Solid-Phase Extraction column for characterize metals, and elution through two different type of polymers (XAD and DPA) with different affinity for polar and nonpolar organic pollutants. Chemical analyses concerned determination of ammonia and trace elements in the untreated sample and after manipulation intended to remove or modify bioavailability of ammonia and metals. The "hybrid" Phase I-Phase II TIE sequence proved to be a reliable and effective tool for the identification of main toxicant of concern in a highly toxic and contaminated interstitial water sample, also in presence of high concentration of potential confounding factors (ammonia). The addition of U. rigida was the more reliable treatment for the removal of ammonia, due to the concurrent release of particles and potentially toxic elements, such as Ba, Rb, and Tl by zeolite column, which may increase toxicity in the post-column sample. The combined use of polymers with different affinity for the various classes of organic pollutants was essential to identify the contribution of polar organic compounds to the observed toxicity.

Dual Analysis of Virus-Host Interactions: The Case of Ostreid herpesvirus 1 and the Cupped Oyster Crassostrea gigas

Dual analyses of the interactions between Ostreid herpesvirus 1 (OsHV-1) and the bivalve Crassostrea gigas during infection can unveil events critical to the onset and progression of this viral disease and can provide novel strategies for mitigating and preventing oyster mortality. Among the currently used “omics” technologies, dual transcriptomics (dual RNA-seq) coupled with the analysis of viral DNA in the host tissues has greatly advanced the knowledge of genes and pathways mostly contributing to host defense responses, expression profiles of annotated and unknown OsHV-1 open reading frames (ORFs), and viral genome variability. In addition to dual RNA-seq, proteomics and metabolomics analyses have the potential to add complementary information, needed to understand how a malacoherpesvirus can redirect and exploit the vital processes of its host. This review explores our current knowledge of “omics” technologies in the study of host-pathogen interactions and highlights relevant applications of these fields of expertise to the complex case of C gigas infections by OsHV-1, which currently threaten the mollusk production sector worldwide.

Shifting Balance of Protein Synthesis and Degradation Sets a Threshold for Larval Growth Under Environmental Stress.

Exogenous environmental factors alter growth rates, yet information remains scant on the biochemical mechanisms and energy trade-offs that underlie variability in the growth of marine invertebrates. Here we study the biochemical bases for differential growth and energy utilization (as adenosine triphosphate [ATP] equivalents) during larval growth of the bivalve Crassostrea gigas exposed to increasing levels of experimental ocean acidification (control, middle, and high pCO2, corresponding to ∼400, ∼800, and ∼1100 µatm, respectively). Elevated pCO2 hindered larval ability to accrete both shell and whole-body protein content. This negative impact was not due to an inability to synthesize protein per se, because size-specific rates of protein synthesis were upregulated at both middle and high pCO2 treatments by as much as 45% relative to control pCO2. Rather, protein degradation rates increased with increasing pCO2. At control pCO2, 89% of cellular energy (ATP equivalents) utilization was accounted for by just 2 processes in larvae, with protein synthesis accounting for 66% and sodium-potassium transport accounting for 23%. The energetic demand necessitated by elevated protein synthesis rates could be accommodated either by reallocating available energy from within the existing ATP pool or by increasing the production of total ATP. The former strategy was observed at middle pCO2, while the latter strategy was observed at high pCO2. Increased pCO2 also altered sodium-potassium transport, but with minimal impact on rates of ATP utilization relative to the impact observed for protein synthesis. Quantifying the actual energy costs and trade-offs for maintaining physiological homeostasis in response to stress will help to reveal the mechanisms of resilience thresholds to environmental change.

First data on three bivalve species exposed to an intra-harbour polymetallic contamination (La Rochelle, France)

Evaluating diffuse sediment contamination in the environment is a major concern with the aim of reaching a good chemical and ecological state of the littoral zone. In this study the risks of chronic chemical contamination and consequences in the bivalves Crassostrea gigas, Mytilus sp. and Mimachlamys varia were evaluated in coastal environments. The objective here was to understand the anthropological phenomena that affect the functioning of the marina of La Rochelle (semi-closed environment). Harbours seeking ecomanagement accreditations (such as the international reference ISO 14001) constitute zones of interest to implement biomonitoring studies. The biological effects of chemical pollution in the Marina of La Rochelle were studied to develop a multi-biomarker biomonitoring approach on specific marine species of this site. Moreover, a genetic (DNA barcoding) approach was applied to validate the species identity of collected bivalves. Of the three species tested the scallop, M. varia, was the most sensitive to metal exposure.

Serum amyloid A in marine bivalves: An acute phase and innate immunity protein

Serum amyloid A (SAA) is among the most potent acute phase proteins (APP) in vertebrates. After injury, its early expression can dramatically increase to promote the recruitment of immuno-competent cells, expression of pro-inflammatory proteins and the activation of the innate immune defences. Although APP have been studied in many vertebrates, only recently their search was extended to invertebrates and the finding of SAA-like molecules has opened new questions on the immune-regulatory functions of these soluble proteins in the animal kingdom. Taking advantage of the considerable amount of genomic and transcriptomic data currently available, we retrieved 51 SAA-like proteins in several protostome taxa comprising 21 marine bivalve species and basal metazoans. In addition to vertebrate-like SAAs, we identified a second protein type with peculiar features. In the bivalves Crassostrea gigas and Mytilus galloprovincialis, both digital expression analysis and qPCR data indicated an induction of the classical SAA after bacterial challenge.

Understanding the dynamics of δ 13C and δ 15N in soft tissues of the bivalve Crassostrea gigas facing environmental fluctuations in the context of Dynamic Energy Budgets (DEB)

We studied the dynamics of stable isotopes δ 13C and δ 15N of an opportunistic suspension feeder the Pacific oyster ( Crassostrea gigas) to better understand the factors that influence the trophic enrichment (trophic-shift, Δ) between primary producers and consumers. Most of the previous studies on this topic do not quantify mass fluxes or isotopic discrimination phenomena in the organism, which are two pillars in isotope ecology. We used a Dynamic Energy Budget (DEB) approach ( Kooijman, 2010) to quantify i) the fluxes of elements and isotopes in C. gigas soft tissues and ii) the impact of the scaled feeding level, the organism mass and the isotopic ratio of food on the “trophic-shift” Δ, and isotope turnover in tissues. Calibration and parametrization modeling were based on data from the literature. We showed that a five-fold increase in scaled feeding level leads to a decrease of the trophic-shift value of 35% for carbon and 43% for nitrogen. This can be explained by the molecule selection for the anabolic and/or catabolic way. When f increases due to the reserve dynamic formulation in the standard DEB model, the half-life of the isotopic ratio t δ 1/2 in tissues also decreases from 13.1 to 7.9 d for δ 13C and from 22.1 to 10.3 d for δ 15N. Organism mass also affects the trophic-shift value: an increase of the individual initial mass from 0.025 g to 0.6 g leads to an enrichment of 22% for δ 13C and 21% for δ 15N. For a large individual, these patterns show that a high structural volume has to be maintained. Another consequence of the mass effect is an increase of the half-life for δ 13C from 6.6 to 12.0 d, and an increase of the half life for δ 15N from 8.3 to 19.4 d. In a dynamic environment, the difference in the isotopic ratios between the individual tissues and the food (δ 13C W –δ 13C X ) exhibits a range of variation of 2.02‰ for carbon and 3.03‰ for nitrogen. These results highlight the potential errors in estimating the contributions of the food sources without considering the selective incorporation of isotopes. We conclude that the Dynamic Energy Budget model is a powerful tool to investigate the fate of isotopes in organisms.

Molecular cloning of a new member of the p53 family from the Pacific oyster Crassostrea gigas and seasonal pattern of its transcriptional expression level

Like other sessile filter-feeding molluscs, oysters may be exposed in the natural environment to a variety of contaminants. Long-term exposure to pollutants may be one factor affecting prevalence of cancerous-like disorders, such as neoplasia. Environmentally induced alterations in p53 protein expression, in relation to leukemia, have been reported in various mollusc species inhabiting polluted water, suggesting that p53 proteins can also be used as a marker for environmental research. This work reports the cloning and sequencing of a p53-like cDNA in the mollusc bivalve Crassostrea gigas. The deduced amino acid sequences of p53 shared a high degree of homology with the homologues from other mollusc species, including typical eukaryotic p53 signature sequences. We examined the p53 transcription expression pattern during the annual cycle in oyster gills and whole soft tissues in four locations along the French coasts. Real-time PCR analysis suggested that strong variations at p53 mRNA level are probably synchronized with the seasonal cycle at the four locations investigated.

Impact of seston characteristics on qualitative particle selection sites and efficiencies in the pseudolamellibranch bivalve Crassostrea gigas

To date, knowledge of the qualitative particle selection sites and conditions in the widely-distributed bivalve Crassostrea gigas is incomplete, having focussed either on heterogeneous particles, or on particles intentionally too large to enter the gill principal filament tracts. We used endoscope-directed sampling and the intact diatom-empty, cleaned frustule approach to unambiguously establish qualitative selection sites and the influence of seston quality (varying proportions of intact diatoms and empty, cleaned frustules) and quantity (particle loads) on the degree of qualitative selection. Normally-feeding oysters were presented test mixtures of the naturally-occurring Actinoptychus senarius (small enough to enter the gill principal filaments), and the potential selection sites (gill: dorsal and ventral collecting tracts; labial palps: anteriorly-deposited pseudofaeces), were sampled for comparison with the proportions and concentrations of the ambient medium. Qualitative selection was demonstrated at both the gills and labial palps. Gill selection efficiency was shown to be directly proportional to seston quality and quantity, using a technique independant of pseudofeces mucus content. The oyster gill is thus able to increase ingested food quality when environmental food quality is low and / or when seston concentrations are high, which is typical of oyster habitats. Palp selection efficiency was directly proportional to seston quality, but at the highest concentration tested, no qualitative selection was observed on the labial palps, probably due to overload on these smaller organs. The partial functional redundancy of these key processing organs in heterorhabdic species such as oysters and scallops may enhance their success in high-turbidity habitats.

Seasonal variation in weight and biochemical composition of the Pacific oyster, Crassostrea gigas in relation to the gametogenic cycle and environmental conditions of the Bizert lagoon, Tunisia

Seasonal variations in biochemical composition of the bivalve Crassostrea gigas were studied from February 2002 until January 2003 in the Bizert lagoon in relation to environmental conditions and reproductive events. Separate analyses were made of gonad-visceral mass. The gametogenic cycle comprised tow phases: gametogenesis including ripeness and spawning (March–September) and sexual repose during the rest of the year. The C. gigas in the Bizert lagoon showed a clear cycle of energy storage and utilization. Total biochemical composition was about 6.50% ± 3.01 of dry weight in glycogen, 14.34% ± 3.60 of dry weight in protein and 6.67% ± 1.34 of dry weight in lipids. Lipid and glycogen concentrations in the gonad-visceral mass were inversely related: maximum concentrations of glycogen occurred between December and February and corresponded with minimum concentrations of lipids. High glycogen contents recorded in winter supported the processes of gamatogenesis and then decreased progressively to attain low values at maturity and during summer corresponded to the first emission of gametes (June) and the beginning of total spawning, however lipids are accumulated in gonads during the period of maximum ripeness (spring) and decrease in summer. During autumn, the increase of total fatty acids values appeared to be related to the available food in association with an increase in the chlorophyll a concentrations. Protein contents show many variations throughout the year. High protein levels occurred in May and in July which corresponded to oocyte maturation and then decreased in late summer (beginning of total spawning). At the expense of the available food in autumn, protein concentrations increase again to assure recovery of the tissue weight of the bivalve, which induced an increase in the condition indices values.

Physiological bases of genetically determined variation in growth of marine invertebrate larvae: A study of growth heterosis in the bivalve Crassostrea gigas

Many species of marine animals have larval stages whose rates of growth in the plankton are regulated by complex combinations of biological and environmental factors. In this study, we focus on the physiological bases that underlie endogenous variation in growth potential of larvae. Our approach was based on experimental crosses of gravid adults from pedigreed families of the Pacific oyster, Crassostrea gigas. This produced large numbers of larvae with different growth rates when reared under similar environmental conditions of food and temperature. A total of 35 larval families were reared to test hypotheses regarding the physiological bases of growth variation. Growth rate of these larval families varied over a five-fold range, from 3.4 (± 0.5, S.E.) to 17.6 (± 0.6) μm day − 1 . The suite of integrated measurements applied to study growth variation included size, biochemical compositions, rates of particulate and dissolved nutrient acquisition, absorption efficiencies, respiration rates and enzyme activities. We show that a complex set of physiological processes regulated differences in genetically determined growth rates of larvae. One-half of the energy required for faster growth came from an enhanced, size-specific feeding ability. Differences in absorption rates were not significant for slow- and fast-growing larvae, nor were differences in size-specific respiration rates. Metabolic processes accounted for the additional 50% of the energy “savings” required to explain enhanced growth rates. We propose that different protein depositional efficiencies could account for this energy saving. Quantitative analyses of the endogenous physiological factors that cause variation in growth rate will allow for a more sophisticated understanding of growth, survival and recruitment potential of larvae.

Native Parasites Adopt Introduced Bivalves of the North Sea

Introduced species may have a competitive advantage over native species due to a lack of predators or pathogens. In the North Sea region, it has been assumed that no metazoan parasites are to be found in marine introduced species. In an attempt to test this assumption, we found native parasites in the introduced bivalves Crassostrea gigas and Ensis americanus with a prevalence of 35% and 80%, respectively, dominated by the trematode Renicola roscovita. When comparing these introduced species with native bivalves from the same localities, Mytilus edulis and Cerastoderma edule, trematode intensity was always lower in the introduced species. These findings have three major implications: (1) introduced bivalves are not free of detrimental parasites which raises the question whether introduced species have an advantage over native species after invasion, (2) introduced bivalves may divert parasite burdens providing a relief for native species and (3) they may affect parasite populations by influencing the fate of infectious stages, ending either in dead end hosts, not being consumed by potential final hosts or by adding new hosts. Future studies should consider these implications to arrive at a better understanding of the interplay between native parasites and introduced hosts.

Evolution of the arginine kinase gene family

Arginine kinase (AK), catalyzing the reversible transfer of phosphate from MgATP to arginine yielding phosphoarginine and MgADP, is widely distributed throughout the invertebrates and is also present in certain protozoa. Typically, these proteins are found as monomers targeted to the cytoplasm, but true dimeric and contiguous dimeric AKs as well as mitochondrial AK activities have been observed. In the present study, we have obtained the sequences of the genes for AKs from two distantly related molluscs—the cephalopod Nautilus pompilius and the bivalve Crassostrea gigas. These new data were combined with available gene structure data (exon/intron organization) extracted from EST and genome sequencing project databases. These data, comprised of 23 sequences and gene structures from Protozoa, Cnidaria, Platyhelminthes, Mollusca, Arthropoda and Nematoda, provide great insight into the evolution and divergence of the AK family. Sequence and phylogenetic analyses clearly show that the AKs are homologous having arisen from some common ancestor. However, AK gene organization is highly divergent and variable. Molluscan AK genes typically have a highly conserved six-exon/five-intron organization, a structure that is very similar to that of the platyhelminth Schistosoma mansoni Arthropod and nematode AK genes have fewer introns, while the cnidarian and protozoan genes each display unique exon/intron organization when compared to the other AK genes. The non-conservative nature of the AK genes is in sharp contrast to the relatively high degree of conservation of intron positions seen in a homologous enzyme creatine kinase (CK). The present results also show that gene duplication and subsequent fusion events forming unusual two-domain AKs occurred independently at least four times as these contiguous dimers are present in Protozoa, Cnidaria, Platyhelminthes and Mollusca. Detailed analyses of the amino acid sequences indicate that two AKs (one each from Drosophila and Caenorhabditis) have what appear to be N-terminal mitochondrial targeting sequences, providing the first evidence for true mitochondrial AK genes. The AK gene family is ancient and the lineage has undergone considerable divergence as well as multiple duplication and fusion events.

Extracellular and Intracellular Acid‐Base Status with Regard to the Energy Metabolism in the OysterCrassostrea gigasduring Exposure to Air

The acid-base status of extra- and intracellular fluids was studied in relation to the anaerobic energy metabolism in the adductor muscle, mantle, gills, and heart of the marine bivalve Crassostrea gigas after exposure to air for periods of 2, 4, 8, 12, 24, and 48 h. Such exposure was found to cause a significant reduction in the pH in the hemolymph (pH(e)) within the first 4 h. The decrease in the pHe was accompanied by elevated Pco2 values, causing [HCO3-] to rise (respiratory acidosis). Thereafter, the pHe fell at a lower rate, and this fall was partially compensated for by a further increase in [HCO3-] in the hemolymph. The increase in the [Ca] levels in the hemolymph indicates a mobilization of Ca2+ from CaCO3 and the involvement of bicarbonates in the buffering of pHe. The main anaerobic end-products that accumulated in the tissues during the first stages of anaerobiosis were alanine and succinate, at a ratio of about 2 : 1. Later on, propionate and acetate were also accumulated at significant rates. In contrast to the adductor muscle, gills, and mantle, opine production in the heart was significant after 12-24 h of exposure to air. Determination of intracellular pH (pHi) revealed that there is a close relationship between the rate of anaerobic end-product accumulation and the extent of intracellular acidosis in the adductor muscle, mantle, and gills. On the contrary, accumulation of anaerobic end-products in the heart did not cause any significant change in its pHi. The intracellular nonbicarbonate, nonphosphate buffering value (beta (NB,NPi)) was determined to be higher in the heart than in the other three tissues and thus probably plays a crucial role in stabilizing heart pHi during exposure to air.

Seasonal Dynamics of Pelagic Larvae and Settling of the Fouling Organisms in Conditions of Thermal Pollution

Year-round comprehensive study of fouling of the cooling system of the Vladivostok Heat Power Plant (VTETs-2) was carried out. The seasonal dynamics of pelagic larvae, their settling, and succession of fouling communities on test plates were analyzed in the intake scoop (Ussuriiskii Bay) and in Zolotoi Rog Bay (Amurskii Bay), which are subjected to thermal pollution by dump waters of VTETs-2. It was shown that the barnacles Balanus crenatus, B. improvisus, and B. amphitrite; bivalves Mytilus trossulus and Crassostrea gigas; polychaetes Polydora limicola and Hydroides ezoensis; and ascidians Molgula manhattensis and Diplosoma mitsukurii were background species in the plate fouling. A high correlation between the seasonal dynamics of larval plankton and the settling of spat was recorded only in some species. The total larval density in Zolotoi Rog Bay, despite the critical level of its pollution, more than 4 times exceeded that of the relatively clean Ussuriiskii Bay. The number of recorded species on the test plates was 2 times higher in Ussuriiskii Bay than in Zolotoi Rog Bay; however, the biomass and population density of settled spat of the dominant forms was several times lower in Ussuriiskii Bay. Thanks to thermal pollution, Zolotoi Rog Bay is an intermediate step for the introduction and acclimation of tropical species, such as B. amphitrite and M. manhattensis, brought by long-distance vessels. Measures aimed at the prevention of intensive settling of mytilids in the cooling system of VTETs-2 are suggested.