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This study comprehensively characterised a protected designation of origin mussel 'Cozza di Scardovari' (Mytilus galloprovincialis) by examining how it is affected by the farming site (outer vs. inner area of the lagoon) and harvest time (21 April vs. 18 May vs. 16 June). Harvest time affected the marketable traits and fatty acid profile of mussels, whereas farming site scarcely affected marketable traits and mussel yields. Mussels from the inner area of the lagoon displayed a superior nutritional profile, including higher contents of proteins (7.8% vs. 7.4%; P < 0.05), lipids (1.2% vs. 1.0%; P < 0.001) and essential amino acids such as tryptophan (+24%; P < 0.05) and valine (+8%; P < 0.05), with a more favourable n-3/n-6 ratio (7.7 vs. 7.0; P < 0.001) than those from the close-to-sea area. Volatile organic compounds, mainly octanoic acid, dimethyl sulphide and 1-penten-3-ol, differed between farming sites within the same lagoon., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Understanding the developmental processes and signaling pathways involved in larval myogenesis and metamorphosis is crucial for comprehending the life history and adaptive strategies of marine organisms. In this study, we investigated the temporal and spatial patterns of myogenesis in the mussel Mytilus coruscus (Mc), focusing on the emergence and transformation of major muscle groups during different larval stages. We also explored the role of the Hedgehog (Hh) signaling pathway in regulating myogenesis and larval metamorphosis. The results revealed distinct developmental stages characterized by the emergence of specific muscular components, such as velum retractor muscles and anterior adductor muscles, in D-veliger and umbo larvae, which are responsible for the planktonic stage. In the pediveliger stage, posterior ventral, posterior adductor, and foot muscles appeared. After larval metamorphosis, the velum structure and its corresponding retractor muscles degenerate, indicating the transition from planktonic to benthic life. We observed a conserved pattern of larval musculature development and revealed a high degree of conservation across bivalve species, with comparable emergence times during myogenesis. Furthermore, exposure to the Hh signaling inhibitor cyclopamine impaired larval muscle development, reduced larval swimming activity, and inhibited larval metamorphosis in M. coruscus. Cyclopamine-mediated inhibition of Hh signaling led to reduced expression of four key genes within the Hh signaling pathway (McHh, McPtc, McSmo, and McGli) and the striated myosin heavy chain gene (McMHC). It is hypothesised that the abnormal larval muscle development in cyclopamine-treated groups may be an indirect effect due to disrupted McMHC expression. We provide evidence for the first time that cyclopamine treatment inhibited larval metamorphosis in bivalves, highlighting the potential involvement of Hh signaling in mediating larval muscle development and metamorphosis in M. coruscus. The present study provides insights into the dynamic nature of myogenesis and the regulatory role of the Hh signaling pathway during larval development and metamorphosis in M. coruscus. The results obtained in this study contribute to a better understanding of the evolutionary significance of Hh signaling in bivalves and shed light on the mechanisms underlying larval muscle development and metamorphosis in marine invertebrates., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Bacteria induced metamorphosis observed in nearly all marine invertebrates. However, the mechanism of bacteria regulating the larvae-juvenile metamorphosis remains unknown. Here, we test the hypothesis that c-di-GMP, a ubiquitous bacterial second-messenger molecule, directly triggers the mollusc Mytilus coruscus larval metamorphosis via the stimulator of interferon genes (STING) receptor. We determined that the deletion of c-di-GMP synthesis genes resulted in reduced c-di-GMP levels and biofilm-inducing activity on larval metamorphosis, accompanied by alterations in extracellular polymeric substances. Additionally, c-di-GMP extracted from tested varying marine bacteria all exhibited inducing activity on larval metamorphosis. Simultaneously, through pharmacological and molecular experiments, we demonstrated that M. coruscus STING (McSTING) participates in larval metamorphosis by binding with c-di-GMP. Our findings reveal that new role of bacterial c-di-GMP that triggers mussel larval metamorphosis transition, and extend knowledge in the interaction of bacteria and host development in marine ecosystems., (© 2024. The Author(s).)

Understanding past coastal variability is valuable for contextualizing modern changes in coastal settings, yet existing Holocene paleoceanographic records for the North American Pacific Coast commonly originate from offshore marine sediments and may not represent the dynamic coastal environment. A potential archive of eastern Pacific Coast environmental variability is the intertidal mussel species Mytilus californianus. Archaeologists have collected copious stable isotopic (δ18O and δ13C) data from M. californianus shells to study human history at California's Channel Islands. When analyzed together, these isotopic data provide windows into 9000 years of Holocene isotopic variability and M. californianus life history. Here we synthesize over 6000 δ18O and δ13C data points from 13 published studies to investigate M. californianus shell isotopic variability across ontogenetic, geographic, seasonal, and millennial scales. Our analyses show that M. californianus may grow and record environmental information more irregularly than expected due to the competing influences of calcification, ontogeny, metabolism, and habitat. Stable isotope profiles with five or more subsamples per shell recorded environmental information ranging from seasonal to millennial scales, depending on the number of shells analyzed and the resolution of isotopic subsampling. Individual shell profiles contained seasonal cycles and an accurate inferred annual temperature range of ~ 5°C, although ontogenetic growth reduction obscured seasonal signals as organisms aged. Collectively, the mussel shell record reflected millennial-scale climate variability and an overall 0.52‰ depletion in δ18Oshell from 8800 BP to the present. The archive also revealed local-scale oceanographic variability in the form of a warmer coastal mainland δ18Oshell signal (-0.32‰) compared to a cooler offshore islands δ18Oshell signal (0.33‰). While M. californianus is a promising coastal archive, we emphasize the need for high-resolution subsampling from multiple individuals to disentangle impacts of calcification, metabolism, ontogeny, and habitat and more accurately infer environmental and biological patterns recorded by an intertidal species., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Vriesman et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Bacteria from coast seawaters are widely known to induce larval recruitment of many invertebrates. However, whether and how deep-sea bacteria, that play crucial roles in the ecological and biogeochemical cycles, promote larval recruitment remains little known. Here, the interaction between deep-sea bacterial biofilms (BFs) and Mytilus coruscus larvae was tested. All these nine deep-sea bacterial isolates triggered planktonic-sessile transition, and the highest percentage of post-larvae was observed in Virgibacillus sp. 1 BF. Except for Pseudomonas sp. 3, Pseudoalteromonas sp. 32 and Bacillus sp. 13, other BF cell  densities were significantly related to their corresponding inductive efficiency. The deep-sea Virgibacillus sp. 1 BF's cue that triggers planktonic-sessile transition was uncovered. Treating Virgibacillus sp. 1 BFs through physic-chemical approaches reduced inducing impact and cell survival. The conditioned water collaborated with formalin-fixed Virgibacillus sp. 1 BF hoisted planktonic-sessile transition efficiency in comparison to each one alone. Thus, two signals derived from deep-sea bacteria trigger planktonic-sessile transition in M. coruscus. This finding firstly demonstrates that deep-sea bacteria has good potential for application in the mussel seed production and provides novel insight to clarify the bacteria-mussel interaction.

Over the last decades, antifouling coatings containing biocidal compounds as active ingredients were used to prevent biofouling, and eco-friendly alternatives are needed. Previous research from our group showed that polymethoxylated chalcones and glycosylated flavones obtained by synthesis displayed antifouling activity with low toxicity. In this work, ten new polymethoxylated flavones and chalcones were synthesized for the first time, including eight with a triazole moiety. Eight known flavones and chalcones were also synthesized and tested in order to construct a quantitative structure-activity relationship (QSAR) model for these compounds. Three different antifouling profiles were found: three compounds ( 1b , 11a and 11b ) exhibited anti-settlement activity against a macrofouling species ( Mytilus galloprovincialis ), two compounds ( 6a and 6b ) exhibited inhibitory activity against the biofilm-forming marine bacteria Roseobacter litoralis and one compound ( 7b ) exhibited activity against both mussel larvae and microalgae Navicula sp. Hydrogen bonding acceptor ability of the molecule was the most significant descriptor contributing positively to the mussel larvae anti-settlement activity and, in fact, the triazolyl glycosylated chalcone 7b was the most potent compound against this species. The most promising compounds were not toxic to Artemia salina , highlighting the importance of pursuing the development of new synthetic antifouling agents as an ecofriendly and sustainable alternative for the marine industry.

The microbiota, the host-associated community of microbes, play important roles in health status and whole body homeostasis of all organisms, including marine species. In bivalves, the microbiota composition has been mainly investigated in adults, whereas little information is available during development. In this work, the microbiota composition of the first larval stages of Mytilus galloprovincialis was evaluated by 16S rRNA gene-based profiling, at 24 and 48 hours post fertilization in comparison with those of eggs and sperm. The main genera detected in both larvae (Vibrio, Pseudoalteromonas, Psychrobium, Colwellia) derived from eggs. However, a clear shift in microbiota was observed in developing larvae compared to eggs, both in terms of core microbiome and relative abundance of different genera. The results provide a first insight into the composition of the microbial communities associated with gametes and early larvae of mussels. Moreover, the impact on larval microbiome of estrogenic chemicals that potentially affect Mytilus early development, 17βestradiol-E 2 , Bisphenol A-BPA and Bisphenol F-BPF (10 μg/L), was investigated. Exposure to estrogenic chemicals leads to changes in abundance of different genera, with distinct and common effects depending on the compound and larval stage. Both potential pathogens (Vibrio, Arcobacter, Tenacibaculum) and genera involved in xenobiotic biotransformation (Oleispira, Shewanella) were affected. The effects of estrogenic compounds on larval microbiome were not related to their developmental effects: however, the results address the importance of evaluating the impact of emerging contaminants on the microbiota of marine invertebrates, including larval stages, that are most sensitive to environmental perturbations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Marine biofouling represents a global economic and ecological challenge and few eco-friendly antifouling agents are available. The aim of this work was to establish the proof of concept that a recently synthesized nature-inspired compound (gallic acid persulfate, GAP) can act as an eco-friendly and effective antifoulant when immobilized in coatings through a non-release strategy, promoting a long-lasting antifouling effect. The synthesis of GAP was optimized to provide quantitative yields. GAP water solubility was assessed, showing values higher than 1000 mg/mL. GAP was found to be stable in sterilized natural seawater with a half-life (DT 50 ) of 7 months. GAP was immobilized into several commercial coatings, exhibiting high compatibility with different polymeric matrices. Leaching assays of polydimethylsiloxane and polyurethane-based marine coatings containing GAP confirmed that the chemical immobilization of GAP was successful, since releases up to fivefold lower than the conventional releasing systems of polyurethane-based marine coatings were observed. Furthermore, coatings containing immobilized GAP exhibited the most auspicious anti-settlement effect against Mytilus galloprovincialis larvae for the maximum exposure period (40 h) in laboratory trials. Overall, GAP promises to be an agent capable of improving the antifouling activity of several commercial marine coatings with desirable environmental properties.

Ocean acidification (OA) represents a serious challenge to marine ecosystems. Laboratory studies addressing OA indicate broadly negative effects for marine organisms, particularly those relying on calcification processes. Growing evidence also suggests OA combined with other environmental stressors may be even more deleterious. Scaling these laboratory studies to ecological performance in the field, where environmental heterogeneity may mediate responses, is a critical next step toward understanding OA impacts on natural communities. We leveraged an upwelling-driven pH mosaic along the California Current System to deconstruct the relative influences of pH, ocean temperature, and food availability on seasonal growth, condition and shell thickness of the ecologically dominant intertidal mussel Mytilus californianus. In 2011 and 2012, ecological performance of adult mussels from local and commonly sourced populations was measured at 8 rocky intertidal sites between central Oregon and southern California. Sites coincided with a large-scale network of intertidal pH sensors, allowing comparisons among pH and other environmental stressors. Adult California mussel growth and size varied latitudinally among sites and inter-annually, and mean shell thickness index and shell weight growth were reduced with low pH. Surprisingly, shell length growth and the ratio of tissue to shell weight were enhanced, not diminished as expected, by low pH. In contrast, and as expected, shell weight growth and shell thickness were both diminished by low pH, consistent with the idea that OA exposure can compromise shell-dependent defenses against predators or wave forces. We also found that adult mussel shell weight growth and relative tissue mass were negatively associated with increased pH variability. Including local pH conditions with previously documented influences of ocean temperature, food availability, aerial exposure, and origin site enhanced the explanatory power of models describing observed performance differences. Responses of local mussel populations differed from those of a common source population suggesting mussel performance partially depended on genetic or persistent phenotypic differences. In light of prior research showing deleterious effects of low pH on larval mussels, our results suggest a life history transition leading to greater resilience in at least some performance metrics to ocean acidification by adult California mussels. Our data also demonstrate "hot" (more extreme) and "cold" (less extreme) spots in both mussel responses and environmental conditions, a pattern that may enable mitigation approaches in response to future changes in climate., Competing Interests: The authors have declared that no competing interests exist.

The modern technology brought new engineering materials (e.g. nanostructured materials) with advantageous characteristics such as a high capacity to decontaminate water from pollutants (for example metal(loid)s). Among those innovative materials the synthesis of nanostructured materials (NSMs) based on graphene as graphene oxide (GO) functionalized with polyethyleneimine (GO-PEI) had a great success due to their metal removal capacity from water. However, research dedicated to environmental risks related to the application of these materials is still non-existent. To evaluate the impacts of such potential stressors, benthic species can be a good model as they are affected by several environmental constraints. Particularly, the mussel Mytilus galloprovincialis has been identified by several authors as a bioindicator that responds quickly to environmental disturbances, with a wide spatial distribution and economic relevance. Thus, the present study aimed to evaluate the impacts caused in M. galloprovincialis by seawater previously contaminated by Hg and decontaminated using GO-PEI. For this, histopathological and biochemical alterations were examined. This study demonstrated that mussels exposed to the contaminant (Hg), the decontaminant (GO-PEI) and the combination of both (Hg + GO-PEI) presented an increment of histopathological, oxidative stress and metabolic alterations if compared to organisms under remediated seawater and control conditions The present findings highlight the possibility to remediate seawater with nanoparticles for environmental safety purposes., Competing Interests: Declaration of Competing Interest The Authors whose names are listed immediately below certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers' bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affi liations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Determination of the median effective concentration (EC50) of Cu on Mytilus galloprovincialis larvae by diffusive gradient in thin films (DGT) has been shown to effectively reduce the need to consider dissolved organic carbon (DOC) concentration and quality. A standard toxicity test protocol was used to validate previously modeled protective effects, afforded to highly sensitive marine larvae by ligand competition, in 5 diverse site waters. The results demonstrate significant narrowing of M. galloprovincialis toxicological endpoints, where EC50s ranged from 3.74 to 6.67 μg/L as C DGT Cu versus 8.76 to 26.8 μg/L as dissolved Cu (Cu DISS ) over a DOC range of 0.74 to 3.11 mg/L; Strongylocentrotus purpuratus EC50s were 10.5 to 19.3 μg/L as C DGT Cu versus 22.7 to 67.1 μg/L as Cu DISS over the same DOC range. The quality of DOC was characterized by fluorescence excitation and emission matrices. The results indicate that the heterogeneity of competing Cu binding ligands, in common marine waters, minimizes the need for class determinations toward explaining the degree of protection. Using conservative assumptions, an M. galloprovincialis C DGT Cu EC50 of 3.7 µg/L and corresponding criterion maximum concentration C DGT Cu of 1.8 µg/L, for universal application by regulatory compliance-monitoring programs, are proposed as a superior approach toward both integration of dynamic water quality over effective exposure periods and quantification of biologically relevant trace Cu speciation. Environ Toxicol Chem 2020;39:873-881. © 2020 SETAC., (© 2020 SETAC.)

We tested the hypothesis that environmental trophic conditions prominent during the growing period (nurture conditions) can modify the differing physiological profiles between fast (F)- and slow (S)-growing juveniles of the mussel Mytilus galloprovincialis. Approximately 200 individuals were fed a high organic content diet dosed below the pseudofaeces threshold (BP), whereas another 200 were fed a low organic content diet dosed above the pseudofaeces threshold (AP), forcing them to maintain a continuous production of pseudofaeces. After 3 months, F and S individuals in each rearing condition were selected and used in feeding experiments. We measured the physiological parameters of the energy balance of selected F and S mussels fed on 4 different diets and tested the effects of the rearing condition (BP vs AP) and growth condition (F vs S) upon the physiological variables. Irrespective of the rearing condition, F-mussels attained higher values of scope for growth with the four experimental diets due to their capacity to display higher clearance rates and preingestive selection efficiencies. F-individuals also had higher gill-surface areas than S individuals. We discussed the role of the gills in determining inter-individual growth rate differences in the mussel.

As a stage of life cycle, larval settlement and metamorphosis are critical processes for persistence of many marine invertebrate populations. Bacterial biofilms (BFs) could induce larval settlement and metamorphosis. Pseudoalteromonas, a widely distributed genus of marine bacteria, showed inductive effects on several invertebrates. However, how Pseudoalteromonas BFs induce settlement and metamorphosis of Mytilus coruscus remains unclear. Pseudoalteromonas marina BFs with the highest inducing activity were further investigated to define inductive cues. Surface-bound products of P. marina BFs could induce larval settlement and metamorphosis. P. marina BFs treated with formalin, antibiotics, ultraviolet irradiation, heat and ethanol significantly reduced inductive effects and cell survival rates. The confocal laser scanning microscopy and the biovolume analysis showed the dominance of α-polysaccharides on P. marina BFs. Treatment of BFs with amylases, proteases and lipase led to the decrease of inducing activity, suggesting that inductive cues of P. marina BFs may comprise of molecular domains of polysaccharides, proteins, and lipids. Finding inductive cues of BFs could put forward further studies about the mechanism of larval settlement and metamorphosis of marine invertebrates.

Bivalve biomineralization is a highly complex and organized process, involving several molecular components identified in adults and larval stages. However, information is still scarce on the ontogeny of the organic matrix before calcification occurs. In this work, first shell formation was investigated in the mussel Mytilus galloprovincialis . The time course of organic matrix and CaCO 3 deposition were followed at close times post fertilization (24, 26, 29, 32, 48 h) by calcofluor and calcein staining, respectively. Both components showed an exponential trend in growth, with a delay between organic matrix and CaCO 3 deposition. mRNA levels of genes involved in matrix deposition (chitin synthase; tyrosinase- TYR) and calcification (carbonic anhydrase; extrapallial protein) were quantified by qPCR at 24 and 48 hours post fertilization (hpf) with respect to eggs. All transcripts were upregulated across early development, with TYR showing highest mRNA levels from 24 hpf. TYR transcripts were closely associated with matrix deposition as shown by in situ hybridization. The involvement of tyrosinase activity was supported by data obtained with the enzyme inhibitor N-phenylthiourea. Our results underline the pivotal role of shell matrix in driving first CaCO 3 deposition and the importance of tyrosinase in the formation of the first shell in M. galloprovincialis .

Biofouling is one of the challenges that can strongly affect the finfish farm economy. Although several studies on biofouling in aquaculture have been conducted in the Mediterranean Sea, they focused on specific taxa or were limited to a particular period of sampling. The present study investigated for the first time the development, composition and variation in a biofouling community in a finfish farm with immersion time, season and depth. The results indicate that all these factors influence biofouling succession and recruitment. Moreover, the species that had a crucial role in structuring the community and in the farm cleaning activities were the ascidian Styela plicata and the bivalve Mytilus galloprovincialis . Compared with the literature data, the results highlight the heterogeneity in the composition of the biofouling present in the Mediterranean Sea. Moreover, such knowledge of the biofouling community could provide important information about management efforts and the costs that farmers will face when siting new fish farms.

The present study aims to evaluate the effects of copper and silver alone or along with a moderate temperature increase on embryonic development, DNA integrity and target gene expression levels in early life stages of Mytilus galloprovincialis. For this purpose, upon fertilized embryos were exposed to a sub-lethal concentration of Cu (9.54 μg/L), Ag (2.55 μg/L) and to the mixture of the two metals (Cu (6.67 μg/L) + Ag (1.47 μg/L)) along with a temperature gradient (18, 20 and 22 °C). In all experiments, larvae were exposed to stressors for 48 h except for those designed to DNA damage analysis exposed only for 24 h (before shell formation).Our results showed a significant increase in the percentage of malformed D-larvae (p < 0.05) with increasing temperature and exposure to silver and copper alone or in a mixture. Moreover, metal toxicity increased significantly (p < 0.05) with the temperature rise. Genotoxicity was evaluated using classic and modified with Formamidopyrimidine DNA glycosylase (Fpg) Comet assay. Results suggest that co-exposure to metals and temperature significantly increased DNA damage on mussel larvae with a more accentuated oxidative damage. A significant transcription modulation was observed for genes involved in DNA repair and DNA replication (p53, DNA ligase II and topoisomerase II) when larvae are exposed to a single stressor. However, in the case of multiple stresses, caspase involved in the cell apoptosis pathway was overexpressed. Our study suggests that mussel larvae exposed to a moderate increase in temperature may have a compromised ability to defend against genotoxicity. This is particularly relevant in the context of global warming and thermal pollution., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Toxicity of silver nanoparticles (Ag NPs) to aquatic organisms has been widely studied. However, the potential toxic effects of Ag NPs ingested through the food web, especially at environmentally relevant concentrations, as well as the potential effects on the offspring remain unknown. The aims of this work were to screen the cytotoxicity of Poly N‑vinyl‑2‑pirrolidone/Polyethyleneimine (PVP/PEI) coated 5 nm Ag NPs in hemocytes exposed in vitro and to assess the effects of dietary exposure to Ag NPs on mussels growth, immune status, gonad condition, reproductive success and offspring embryo development. For this, mussels Mytilus galloprovincialis were fed daily with microalgae Isochrysis galbana previously exposed for 24 h to a dose close to environmentally relevant concentrations (1 μg Ag/L Ag NPs) and to a high dose of 10 μg Ag/L Ag NPs. After 24 h of in vitro exposure, Ag NPs were cytotoxic to mussel hemocytes starting at 1 mg Ag/L (LC50: 2.05 mg Ag/L). Microalgae significantly accumulated Ag after the exposure to both doses and mussels fed for 21 days with microalgae exposed to 10 μg Ag/L Ag NPs significantly accumulated Ag in the digestive gland and gills. Sperm motility and fertilization success were not affected but exposed females released less eggs than non-exposed ones. The percentage of abnormal embryos was significantly higher than in control individuals after parental exposure to both doses. Overall, results indicate that Ag NPs taken up through the diet can significantly affect ecologically relevant endpoints such as reproduction success and embryo development in marine mussels., (Copyright © 2018 Elsevier B.V. All rights reserved.)

High latitudes are considered particularly vulnerable to ocean acidification, since they are naturally low in carbonate ions. The edible mussel Mytilus chilensis is a common calcifier inhabiting marine ecosystems of the southern Chile, where culturing of this species is concentrated and where algal blooms produced by the toxic dinoflagellate A. catenella are becoming more frequent. Juvenile Mytilus chilensis were exposed to experimental conditions simulating two environmental phenomena: pCO 2 increase and the presence of paralytic shellfish toxins (PST) produced by the dinoflagellate Alexandrium catenella. Individuals were exposed to two levels of pCO 2 : 380 μatm (control condition) and 1000 μatm (future conditions) over a period of 39 days (acclimation), followed by another period of 40 days exposure to a combination of pCO 2 and PST. Both factors significantly affected most of the physiological variables measured (feeding, metabolism and scope for growth). However, these effects greatly varied over time, which can be explained by the high individual variability described for mussels exposed to different environmental conditions. Absorption efficiency was not affected by the independent effect of the toxic diet; however, the diet and pCO 2 interaction affected it significantly. The inhibition of the physiological processes related with energy acquisition by diets containing PST, may negatively impact mussel fitness, which could have important consequences for both wild and cultured mussel populations, and thus, for socioeconomic development in southern Chile., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Coastal marine ecosystems experience dynamic fluctuations in seawater carbonate chemistry. The importance of this variation in the context of ocean acidification requires knowing what aspect of variability biological processes respond to. We conducted four experiments (ranging from 3 to 22 days) with different variability regimes (pH T 7.4-8.1) assessing the impact of diel fluctuations in carbonate chemistry on the early development of the mussel Mytilus galloprovincialis. Larval shell growth was consistently correlated to mean exposures, regardless of variability regimes, indicating that calcification responds instantaneously to seawater chemistry. Larval development was impacted by timing of exposure, revealing sensitivity of two developmental processes: development of the shell field, and transition from the first to the second larval shell. Fluorescent staining revealed developmental delay of the shell field at low pH, and abnormal development thereof was correlated with hinge defects in D-veligers. This study shows, for the first time, that ocean acidification affects larval soft-tissue development, independent from calcification. Multiple developmental processes additively underpin the teratogenic effect of ocean acidification on bivalve larvae. These results explain why trochophores are the most sensitive life-history stage in marine bivalves and suggest that short-term variability in carbonate chemistry can impact early larval development.

Marine mussel aggregations act as a substratum and refuge for many fouling species. Mussel cultivation in Galicia, Spain, is carried out on hanging ropes in subtidal systems. The fauna associated with this cultivation includes a large number of invertebrates that compete for space or food with the mussels, or use their clusters as a refuge from predators or water turbulence. Outbreaks of the epibiont anemone Actinothoe sphyrodeta have been reported in cultivated Galician mussels since 2013, but their impact has not been investigated rigorously. Here, the temporal and spatial variability of Actinothoe sphyrodeta on mussel shells throughout one year is presented. Sampling of mussel size, weight and byssus attachment strength allowed mussel tenacity (attachment strength relative to size) to be calculated. A higher presence of Actinothoe sphyrodeta correlated with lower mussel tenacity and greater biomass losses, suggesting that this species could be an economically important biofouling component.

Global warming may accelerate growth and distribution of pathogens influencing aquatic organisms' diseases and human health. Despite the extensive research, the biology, cellular development and life cycle and of Marteilia sp. parasites as well as the influence of parasitic infection on the hosts are not fully understood. The aim of this study was to investigate the effect of Marteilia sp. prevalence and infection intensity on mussels' growth rate and morphometric characteristics under natural conditions in Thermaikos Gulf, a major bivalve production area in Greece, during a five-month growth period. The length, width, height and weight of the infected mussels were significantly lower compared to non-infected and the decrease was proportional to the intensity of mussel infection by the parasite. Moreover, the estimation of allometric relations between length, height, width and weight revealed significantly lower growth of mussel wet weight in relation to shell length for infected mussels compared to healthy ones. The negative effect of marteiliosis on the shell length growth rate of infected mussels was also confirmed by von Bertalanffy equations., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Determining the magnitude and causes of intrinsic variability is a main issue in the analysis of bivalve growth. Inter-individual variability in bivalve growth has been attributed to differences in the physiological performance. This hypothesis has been commonly tested comparing the physiological rates of fast and slow growers after size differentiation has occurred. This experimental design may detect a link between growth and physiological performance, but we cannot interpret the posterior physiological performance as a driver for the prior growth variability. Considering these limitations, this work introduces a new methodological framework for the analysis of bivalve growth variability. We have conducted sequential measurements of size and physiological performance (feeding, digestion and metabolic rates) in even-sized mussels growing under homogeneous environmental conditions. This experimental design allows us to distinguish between changes over time within individuals, i.e. growth and trends in the physiological rates, from differences between individuals with respect to a baseline level. In addition, Functional Data Analysis provides powerful tools to summarize all the information obtained in the exhaustive sampling scheme and to test whether differences in the physiological performance enhance growth dispersion. Our results report an increasing dispersion in both size and physiological performance over time. Although mussels grew during the experiment, it is difficult to detect any increasing or decreasing temporal pattern in their feeding, digestion and metabolic rates due to the large inter-individual variability. Comparison between the growth and physiological patterns of mussels with final size above (fast growers) and below (slow growers) the median found that fast growers had larger feeding and digestion rates and lower metabolic expenditures during the experimental culture than mussels with slow growth, which agrees with the hypothesis of a physiological basis for bivalve growth variability., Competing Interests: The authors have declared that no competing interests exist.

Bivalves are worldwide sentinels of anthropogenic pollution. The inclusion of biomarker responses in chemical monitoring is a recommended practise that has to overcome some difficulties. One of them is the time frame between sample collection and sample processing in order to ensure the preservation of enzymatic activities. In the present study, three bivalve species of commercial interest (mussel, Mytilus galloprovincialis, razor shell, Solen marginatus, and cockle, Cerastoderma edule) were processed within <2 h after being retrieved from their natural habitat, and 24 h after being transported in air under cold conditions (6-8 °C) to laboratory facilities. The enzymatic activities were compared in the three species submitted to both conditions revealing no differences in terms of carboxylesterase dependent activities (CEs) using different substrates: p-nitrophenyl acetate (pNPA), p-nitrophenyl butyrate (pNPB), 1-naphthyl acetate (1-NA), 1-naphthyl butyrate (1-NB) and 2-naphthyl acetate (2-NA). In mussels, three tissues were selected (haemolymph, gills and digestive gland). For comparative purposes, in razor shell and cockle only digestive gland was considered as it is the main metabolic organ. Baseline enzymatic activities for CEs were characterised in the digestive gland of the three bivalves using four out of the five selected CE substrates as well as the kinetic parameters (V max and K m ) and catalytic efficiency. The in vitro sensitivity to the organophosphorus metabolite chlorpyrifos oxon was also calculated. IC 50 values (pM-nM range) were lower than those obtained for vertebrate groups which suggest that bivalves have high protection efficiency against this pesticide as well as species dependent particularities., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Intertidal species are frequently exposed to environmental changes associated with multiple stressors, which they must either avoid or tolerate by developing physiological and biochemical strategies. Some of the natural environmental changes are related with the tidal cycle which forces organisms to tolerate the differences between an aquatic and an aerial environment. Furthermore, in these environments, organisms are also subjected to pollutants from anthropogenic sources. The present study evaluated the impacts in Mytilus galloprovincialis exposed to multi-walled carbon nanotubes (0.01 mg/L MWCNTs) when continuously submersed or exposed to tides (5 h of low tide, 7 h of high tide) for 14 days. Our results demonstrated that mussels were physiologically and biochemically affected by MWCNTs, especially when exposed to tides. In fact, when only exposed to the carbon nanoparticles or only exposed to tides, the stress induced was not enough to activate mussels' antioxidant defenses which resulted in oxidative damage. However, when mussels were exposed to the combination of tides and MWCNTs increased metabolism was observed, associated with a possible higher production of reactive oxygen species (ROS), leading to a significant increase in the activities of antioxidant enzymes (superoxide dismutase, SOD and glutathione peroxide, GPx) and oxidized glutathione content (GSSG), preventing the occurrence of cellular damage, expressed as no lipid peroxidation (LPO) or protein carbonylation (PC). Therefore, organisms seemed to be able to tolerate MWCNTs and air exposure during tidal regime; however, the combination of both stressors induced higher oxidative stress. These findings indicate that the increasing presence of carbon nanoparticles in marine ecosystems can induce higher toxic impacts in intertidal organisms compared to organisms continuously submerged. Also, our results may indicate that air exposure can act as a cofounding factor on the assessment of different stressors in organisms living in coastal systems., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Biofouled debris from the 2011 Great East Japan earthquake and tsunami has landed in the Northeast Pacific and along the Hawaiian Islands since 2012. As of 2017, >630 biofouled debris items with >320 living species of algae, invertebrates, and fish have been examined. The invasive mussel Mytilus galloprovincialis was present on >50% of those items. Size, reproduction, and growth of this filter-feeding species were examined to better understand long-distance rafting of a coastal species. The majority of mussels (79%) had developing or mature gametes, and growth rates averaged 0.075±0.018 SE mm/day. Structural and elemental (barium/calcium) analysis of mussel shells generated estimates of growth in coastal waters (mean=1.3 to 25mm total length), which provides an indication of residence times in waters along North America and the Hawaiian Islands prior to landing. Detailed studies of individual species contribute to our understanding of debris as a transport vector and aid efforts to evaluate potential risks associated with marine debris., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this work, we describe results of the reproductive health monitoring studies in Mytilus galloprovincialis following spermatozoa hsp70 expression and protamine-like protein properties. Mussels control (ctr) were released within cages for 30 days in three different marine sites near Naples (Campania, Italy): Bagnoli south (BAs) and Bagnoli north (BAn), both close to a disposal metallurgical factory and in Capo Miseno (CM). Studies of hsp70 gene expression carried out, by RT-qPCR, in mussel spermatozoa have shown varied expression levels, particularly 5, 13, and 15-fold more than ctr in CM, BAs, and BAn, respectively, indicating highest involvement of stress proteins in spermatozoa of mussels in Bagnoli. In order to evaluate the possible risk on Mytilus galloprovincialis sustainability loss, electrophoretic analyses were performed on protamine-like proteins (PL) of collected spermatozoa. The results showed that CM PL were apparently unaltered with respect to ctr PL, while BAs and BAn PL appeared in part in the form of peptides and in part as bands with low mobility. Further, CM and BAs PL showed, by electrophoretic mobility shift assay, a decrease in DNA binding ability and a change in their DNA binding mode. The results of this investigation show the usefulness of the study of alterations of spermatozoa hsp70 expression and protamine-like protein properties for eco-toxicological evaluation using Mytilus galloprovincialis as a bioindicator.

Complex life cycles characterized by uncertainty at transitions between larval/juvenile and adult environments could favour irreversible physiological plasticity at such transitions. To assess whether thermal tolerance of intertidal mussels ( Mytilus californianus ) adjusts to post-settlement environmental conditions, we collected juveniles from their thermally buffered microhabitat from high- and low-shore locations at cool (wave-exposed) and warm (wave-protected) sites. Juveniles were transplanted to unsheltered cages at the two low sites or placed in a common garden. Juveniles transplanted to the warm site for one month in summer had higher thermal tolerance, regardless of origin site. By contrast, common-garden juveniles from all sites had lower tolerance indistinguishable from exposed site transplants. After six months in the field plus a common garden period, there was a trend for higher thermal tolerance at the protected site, while reduced thermal tolerance at both sites indicated seasonal acclimatization. Thermal tolerance and growth rate were inversely related after one but not six months; protected-site transplants were more tolerant but grew more slowly. In contrast to juveniles, adults from low-shore exposed and protected sites retained differences in thermal tolerance after common garden treatment in summer. Both irreversible and reversible forms of plasticity must be considered in organismal responses to changing environments., (© 2018 The Author(s).)

In vivo confocal Raman microscopy (CRM), polarized light microscopy and Fourier transform infrared spectroscopy (FTIR) were used to determine if a significant amount of amorphous calcium carbonate (ACC) exists within larval shells of Baltic mytilid mussels ( Mytilus edulis -like) and whether the amount of ACC varies during larval development. No evidence for ACC was found from the onset of shell deposition at 21 h post-fertilization (hpf) until 48 hpf. Larval Mytilus shells were crystalline from 21 hpf onwards and exhibited CRM and FTIR peaks characteristic of aragonite. Prior to shell deposition at 21 hpf, no evidence for carbonates was observed through in vivo CRM. We further analysed the composition of larval shells in three other bivalve species, Mercenaria mercenaria , Crassostrea gigas and Crassostrea virginica and observed no evidence for ACC, which is in contrast to previous work on the same species. Our findings indicate that larval bivalve shells are composed of crystalline aragonite and we demonstrate that conflicting results are related to sub-optimal measurements and misinterpretation of CRM spectra. Our results demonstrate that the common perception that ACC generally occurs as a stable and abundant precursor during larval bivalve calcification needs to be critically reviewed., (© 2018 The Author(s).)

Carbon capture and storage is a technology that has been widely determined to be one of the best choices for the short-term reduction of atmospheric CO 2 emissions. The aim of this study was to analyze the effects of CO 2 enrichment in the ocean on the mussel species Mytilus galloprovincialis using three different endpoints: mortality, embryo-larval development, and neutral red retention time assays (NRRT). Acute effects were found to be associated with a pH values of 6.0 while citotoxity effects and embryo-larval development were associated with a pH value of 7.0. The NRRT assay and embryo-larval development can be recommended as good endpoints for assessing the environmental risk associated with acidification by CO 2 enrichment because they provide sensitive responses on the effects of changes in seawater pH on mussels in a short period of time. Moreover, this study may support policymakers in finding appropriate solutions for the conservation of marine ecosystems., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The pacific oyster Crassostrea gigas and the Mediterranean mussel Mytilus galloprovincialis are two widely farmed bivalve species which show contrasting behaviour in relation to microbial diseases, with C. gigas being more susceptible and M. galloprovincialis being generally resistant. In a recent study, we showed that different susceptibility to infection exhibited by these two bivalve species may depend on their different capability to kill invading pathogens (e.g., Vibrio spp.) through the action of haemolymph components. Specific microbial-host interactions may also impact bivalve microbiome structure and further influence susceptibility/resistance to microbial diseases. To further investigate this concept, a comparative study of haemolymph and digestive gland 16SrDNA gene-based bacterial microbiota profiles in C. gigas and M. galloprovincialis co-cultivated at the same aquaculture site was carried out using pyrosequencing. Bacterial communities associated with bivalve tissues (hemolymph and digestive gland) were significantly different from those of seawater, and were dominated by relatively few genera such as Vibrio and Pseudoalteromonas. In general, Vibrio accounted for a larger fraction of the microbiota in C. gigas (on average 1.7-fold in the haemolymph) compared to M. galloprovincialis, suggesting that C. gigas may provide better conditions for survival for these bacteria, including potential pathogenic species such as V. aestuarianus. Vibrios appeared to be important members of C. gigas and M. galloprovincialis microbiota and might play a contrasting role in health and disease of bivalve species. Accordingly, microbiome analyses performed on bivalve specimens subjected to commercial depuration highlighted the ineffectiveness of such practice in removing Vibrio species from bivalve tissues.

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., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Multixenobiotic resistance (MXR) efflux transporters constitute a broad-spectrum physiological defense system allowing marine bivalves to cope with environmental challenges. There is, however, scarce information on the type and role that different MXR transporters may have in embryos, which represent the most sensitive stages of bivalves to environmental stress. In this study regulation of MXR-related transporters was investigated in early developmental stages of the Mediterranean mussel (Mytilus galloprovincialis). In vitro fertilization experiments using gametes from naturally-spawning broodstocks were performed to follow embryo development from fertilized eggs (30min post fertilization, pf) to fully developed D-shape veligers (48hpf). Quantitative PCR analyses indicated that ABCB and ABCC transcripts encoding the MXR-related transporters P-glycoproteins (P-gp) and Multidrug resistance proteins (Mrp), respectively, were expressed soon after 30minpf, with ABCC being more expressed than ABCB. Copy numbers of both transcripts were increased in trochophorae and D-veligers. MXR efflux activity assessed using the fluorescent substrate rhodamine 123 and selective P-gp or Mrp inhibitors showed that the P-gp mediated efflux was detected only in D-veligers, while a significant Mrp mediated efflux was detected soon after 30minpf and remained almost unchanged in trochophorae and D-veligers. MXR modulation by propranolol and carbamazepine showed that the pharmaceuticals may act as transcriptional regulators and substrates. Results reported lead to hypothesize that while P-gp aids in xenobiotic efflux performing a prominent protective role, Mrp could be a dual-functioning transporter performing both protective and physiological functions in mussel development., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The increasing usage of nanoparticles has caused their considerable release into the aquatic environment. Meanwhile, anthropogenic CO 2 emissions have caused a reduction of seawater pH. However, their combined effects on marine species have not been experimentally evaluated. This study estimated the physiological toxicity of nano-TiO 2 in the mussel Mytilus coruscus under high pCO 2 (2500-2600 μatm). We found that respiration rate (RR), food absorption efficiency (AE), clearance rate (CR), scope for growth (SFG) and O:N ratio were significantly reduced by nano-TiO 2 , whereas faecal organic weight rate and ammonia excretion rate (ER) were increased under nano-TiO 2 conditions. High pCO 2 exerted lower effects on CR, RR, ER and O:N ratio than nano-TiO 2 . Despite this, significant interactions of CO 2 -induced pH change and nano-TiO 2 were found in RR, ER and O:N ratio. PCA showed close relationships among most test parameters, i.e., RR, CR, AE, SFG and O:N ratio. The normal physiological responses were strongly correlated to a positive SFG with normal pH and no/low nano-TiO 2 conditions. Our results indicate that physiological functions of M. coruscus are more severely impaired by the combination of nano-TiO 2 and high pCO 2 .

In December 2007, >150km of the West coast of Korea were heavily polluted by crude oil leaked from the oil tanker Hebei Spirit, leading to mass mortality of bivalve mollusks on the intertidal areas. Two years after, mussels Mytilus galloprovincialis were collected from two impacted sites to investigate sub-lethal effects of the oil spill. Tissue content in polycyclic aromatic hydrocarbons (PAHs), hemocyte parameters, reproductive status and energetic reserves were analyzed. PAHs in tissues of mussels as well as hemocyte parameters were not different between impacted and control sites. Energetic reserves were altered in mussels from the impacted sites. Glycogen content remained low at polluted sites, whatever the season. Two years after the Hebei Spirit oil spill, mussels then presented altered energetic metabolism. Further investigations are thus warranted to monitor the sustainability of mussel populations on the oil spilled West coast of Korea., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

To understand the potential functions of Wnt7b in different developmental stages and adult tissues of the mussel Mytilus coruscus, the Wnt7b gene was cloned using the rapid amplification of cDNA ends technique. The full-length Wnt7b gene was 1910 bp long, with a 1059-bp open reading frame encoding 352 amino acids. The amino acid sequence of the M. coruscus Wnt7b gene shared high homology with that of Homo sapiens (59%), Mus musculus (61%), Danio rerio (61% similarity), Biomphalaria glabrata (62% similarity), Aplysia californica (66% similarity), and Crassostrea gigas (74%). Wnt7b mRNA expression was detected by reverse transcription PCR in all tissues studied (mantle, adductor muscle, gill, foot, digestive gland, and male and female gonads), with the highest expression found in the gill, and in the male and female gonads. This indicates that Wnt7b may play an important role in gonadal maturation and in the functions of gills in the mussel M. coruscus. Expression of the Wnt7b gene during larval development stages, including the trochophore, D-shaped veliger, umbo veliger, pediveliger, and juvenile stages, was also detected. Wnt7b mRNA was highly expressed in the D-shaped veliger, umbo veliger, and pediveliger larvae stages, suggesting that Wnt7b may participate in larval development and in the process of metamorphosis in the mussel M. coruscus. Taken together, these findings provide new insights into the functions of the Wnt gene family during mussel larval development and settlement and metamorphosis.

Cadmium (Cd) and arsenic (As) are the main metal/metalloid contaminants in the coastal environments of the Bohai Sea, China. In this work, a combined proteomic and metabolomic approach was applied to investigate the biological effects of Cd and As (V) in the early life stage (D-shape larvae) of mussel Mytilus galloprovincialis. Results indicated that Cd was a potential immune toxicant to D-shape larval mussel because of the numerous proteomic responses related to immune system. Additionally, Cd induced oxidative stress, cellular injury and disturbance in nucleic acid metabolism in D-shape larval mussels. However, only two identified proteins were significantly altered in As (V)-treated group, suggesting that D-shape larval mussel was less sensitive to As (V) than to Cd at protein level. These two proteins in response to As (V) suggested that As (V) influenced anti-oxidative system and cell proliferation in D-shape larval mussels. Metabolic responses indicated that Cd and As (V) induced disturbances in osmotic regulation and energy metabolism in D-shape larval mussels via different metabolic pathways. In addition, Cd reduced lipid metabolism as well. This work demonstrated that a combination of proteomics and metabolomics could provide an insightful view in the biological effects of pollutants in mussel M. galloprovincialis at an early life stage., (Copyright © 2016. Published by Elsevier Ltd.)

Cadmium (Cd) has become an important metal contaminant and posed severe risk on the organisms in the coastal environments of the Bohai Sea. Marine mussel Mytilus galloprovincialis is widely distributed along the Bohai coast and consumed as seafood by local residents. Evidences indicate that the early stages of marine organisms are more sensitive to metal contaminants. In this study, we applied two-dimensional electrophoresis-based proteomics to characterize the biological effects of Cd (50 μg L(-1)) in the early life stages (D-shape larval and juvenile) of mussels. The different proteomic responses demonstrated the differential responsive mechanisms to Cd exposure in these two early life stages of mussels. In details, results indicated that Cd mainly induced immune and oxidative stresses in both D-shape larval and juvenile mussels via different pathways. In addition, the significant up-regulation of triosephosphate isomerase and metallothionein confirmed the enhanced energy demand and mobilized detoxification mechanism in D-shape larval mussels exposed to Cd. In juvenile mussels, Cd exposure also induced clear apoptosis. Overall, this work suggests that Cd is a potential immune toxicant to mussel M. galloprovincialis at early life stages., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The harmful alga, Heterocapsa circularisquama, causes recurrent mortalities of bivalve molluscs in Japan, with demonstrated hemolysis and cytotoxicity in rabbit erythrocytes, HeLa cells, and bivalve tissues. Nonetheless, the effects of exposure to sublethal cell densities on the physiological energetics of bivalves have not been investigated, nor the potential involvement of neurotoxicity. In the present study, two sets of experiments were conducted with adult clams, Ruditapes philippinarum. In the first set, the clearance rate (CR), respiration rate (RR), absorption efficiency (AE), ingestion rate (IR), and absorption rate (AR) were examined in clams exposed to H. circularisquama to quantify the scope for growth (SFG) as an indicative of the bioenergetic status of clams (5, 50, 2.5×10(2), and 5×10(2)cellsml(-1); under 15°C and 20°C). In the second set, the activity of the biomarker of neurotoxic exposure, acetylcholinesterase (AChE), was monitored following 3, 6, 24, and 48h of exposure (5, 50, 5×10(2), and 10(3)cellsml(-1), at 20°C) in gills of R. philippinarum, and compared to that in Mediterranean mussels, Mytilus galloprovincialis; a species also affected by H. circualrisquama and in which AChE activity was more extensively studied. At 15°C, CR, IR, and AR were decreased for exposures to 50-5×10(2) cells ml(-1) resulting in a significant decrease in the absorbed energy (A), and a significant decrease in SFG at 5×10(2)cellsml(-1). At 20°C, AE was null for exposures to 2.5×10(2) and 5×10(2)cellsml(-1). RR was decreased at 2.5×10(2) and 5×10(2)cellsml(-1), CR, IR, and AR were decreased at 5-5×10(2)cellsml(-1), and the AE was null for 2.5×10(2)-5×10(2)cellsml(-1) resulting in a significant decrease in the respired energy (R), but mainly in (A) especially at 2.5×10(2) and 5×10(2)cellsml(-1) decreasing the SFG over the entire range of cell density with negative values for 2.5×10(2) and 5×10(2)cellsml(-1). The activity of AChE in both clams and mussels was significantly reduced following 3-48h of exposure to 5-5×10(2)cellsml(-1). Reduction in the energy available for clams to grow and reproduce was induced mainly via decreased energy acquisition, and delayed and/or reduced functions of the digestive organs. In addition, this is the first report of decreased neuroenzymatic activity in two bivalve molluscs induced by exposure to H. circularisquama potentially via either neurotoxic compounds affecting the activity of gill cilia and/or gill muscles, and/or potentially via either neurotoxic compounds affecting the activity of gill cilia and/or gill muscles, and/or non-cholinergic affects associated with other functions; both resulting in decreased SFG. The relationship between the decreased AChE and decreased SFG should be corroborated in future research., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Seawater pH and the availability of carbonate ions are decreasing due to anthropogenic carbon dioxide emissions, posing challenges for calcifying marine species. Marine mussels are of particular concern given their role as foundation species worldwide. Here, we document shell growth and calcification patterns in Mytilus californianus, the California mussel, over millennial and decadal scales. By comparing shell thickness across the largest modern shells, the largest mussels collected in the 1960s-1970s and shells from two Native American midden sites (∼1000-2420 years BP), we found that modern shells are thinner overall, thinner per age category and thinner per unit length. Thus, the largest individuals of this species are calcifying less now than in the past. Comparisons of shell thickness in smaller individuals over the past 10-40 years, however, do not show significant shell thinning. Given our sampling strategy, these results are unlikely to simply reflect within-site variability or preservation effects. Review of environmental and biotic drivers known to affect shell calcification suggests declining ocean pH as a likely explanation for the observed shell thinning. Further future decreases in shell thickness could have significant negative impacts on M. californianus survival and, in turn, negatively impact the species-rich complex that occupies mussel beds., (© 2016 The Author(s).)

The mantle is an organ common to all molluscs and is at the forefront of the biomineralisation process. The present study used the Mediterranean mussel (Mytilus galloprovincialis) as a model species to investigate the structural and functional role of the mantle in shell formation. The transcriptomes of three regions of the mantle edge (umbo to posterior edge) were sequenced using Illumina technology which yielded a total of 61,674,325 reads after adapter trimming and filtering. The raw reads assembled into 179,879 transcripts with an N50 value of 1086bp. A total of 1363 transcripts (321, 223 and 816 in regions 1, 2 and 3, respectively) that differed in abundance in the three mantle regions were identified and putative function was assigned to 54% using BLAST sequence similarity searches (cut-off less than 1e(-10)). Morphological differences detected by histology of the three mantle regions was linked to functional heterogeneity by selecting the top five most abundant Pfam domains in the annotated 1363 differentially abundant transcripts across the three mantle regions. Calcium binding domains dominated region two (middle segment of the mantle edge). Candidate biomineralisation genes were mined and tested by qPCR. This revealed that Flp-like, a penicillin binding protein potentially involved in shell matrix maintenance of the Pacific oyster (Crassostrea gigas), had significantly higher expression in the posterior end of the mantle edge (region one). Our findings are intriguing as they indicate that the mantle edge appears to be a heterogeneous tissue, displaying structural and functional bias., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Polychlorinated biphenyls, organochlorine pesticides and trace metals were determined in tissues of bivalve molluscs (Mytilus galloprovincialis, Ostrea edulis, Venus verrucosa, Arca noae and Callista chione), collected from 11 harvesting and 2 cultured locations along the eastern Adriatic coast, in May and November 2012. Concentrations (ng g(-1) dry weight) of organochlorines ranged from 1.53 to 21.1 for PCBs and 0.68 to 5.21 for p,p'-DDTs. HCB, lindane, heptachlor and aldrin-like compounds were found in lower levels or were not detected. Metal concentrations (mg kg(-1) dry weight) ranged from 0.23 to 4.03 for Cd, 0.87-3.43 for Cr, 3.69-202.3 for Cu, 0.06-0.26 for HgT, 0.62-9.42 for Ni, 0.95-4.64 for Pb, and 55.76-4010.3 for Zn. Established organochlorine and trace metal levels were lower than the maximum allowable levels in seafood set by the European Commission., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Numerous studies on molluscs have been carried out to clarify the physiological roles of haemolymph serum proteins and haemocytes. However, little is known about the presence and functional role of the serum metabolites. In this study, Nuclear Magnetic Resonance (NMR) was used to assess whether changes of the metabolic profile of Mytilus galloprovincialis haemolymph may reflect alterations of the physiological status of the organisms due to environmental stressors, namely copper and temperature. Mussel haemolymph was taken from the posterior adductor muscle after a 4-day exposure to ambient (16 °C) or high temperature (24 °C) and in the absence or presence (5 μg/L, 20 μg/L, or 40 μg/L) of sublethal copper (Cu(2+)). The total glutathione (GSH) concentration in the haemolymph of both control and treated mussels was minimal, indicating the absence of significant contaminations by muscle intracellular metabolites due to the sampling procedure. In the (1)H-NMR spectrum of haemolymph, 27 metabolites were identified unambiguously. The separate and combined effects of exposure to copper and temperature on the haemolymph metabolic profile were assessed by Principal Component Analysis (PCA) and Ranking-PCA multivariate analysis. Changes of the metabolomic profile due to copper exposure at 16 °C became detectable at a dose of 20 μg/L copper. Alanine, lysine, serine, glutamine, glycogen, glucose and protein aliphatics played a major role in the classification of the metabolic changes according to the level of copper exposition. High temperature (24 °C) and high copper levels caused a coherent increase of a common set of metabolites (mostly glucose, serine, and lysine), indicating that the metabolic impairment due to high temperature is enforced by the presence of copper. Overall, the results demonstrate that, as for human blood plasma, the analysis of haemolymph metabolites represents a promising tool for the diagnosis of pollutant-induced stress syndrome in marine mussels., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Inorganic arsenic (As) is a known pollutant including two chemical forms (arsenite (As III) and arsenate (As V)), in marine and coastal environment. Marine mussel Mytilus galloprovincialis is an important environmental monitoring species around the world. In this study, we focused on valence-specific responses of As in juvenile mussel M. galloprovincialis using a combined proteomic and metabolomic approach. Metabolic responses indicated that As (III) mainly caused disturbance in osmotic regulation in juvenile mussels. As (V) caused disturbances in both osmotic regulation and energy metabolism marked by different metabolic responses, including betaine, taurine, glucose and glycogen. Proteomic responses exhibited that As (III) had a significant negative effect on cytoskeleton and cell structure (actin and collagen alpha-6(VI) chain). As (V) affected some key enzymes involved in energy metabolism (cytosolic malate dehydrogenase, cMDH) and cell development (ornithine aminotransferase and astacin). Overall, all these results confirmed the valence-specific responses in juvenile mussels to As exposures. These findings demonstrate that a combined metabolomic and proteomic approach could provide an important insight into the toxicological effects of environmental pollutants in organisms., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The neurotoxin β-N-methylamino-l-alanine (BMAA) is naturally present in some microalgal species in the marine environment. The accumulation of BMAA has widely been observed in filter-feeding bivalves that are known to consume primary producers constituting the base of complex aquatic food webs. This study was performed to assess the occurrence of BMAA and isomers in mollusks collected from nine representative shellfish production areas located on the three French coasts (Channel, Atlantic and Mediterranean sites). The use of a highly selective and sensitive HILIC-MS/MS method, with D5DAB as internal standard, revealed the systematic detection of BMAA and DAB, in concentrations ranging from 0.20 to 6.7 μg g(-1) dry weight of digestive gland tissues of mollusks. While we detected BMAA in four strains of diatoms in a previous study, here BMAA was only detected in one diatom species previously not investigated out of the 23 microalgal species examined (belonging to seven classes). The concentrations of BMAA and DAB in mussels and oysters were similar at different sampling locations and despite the high diversity of phytoplankton populations that mollusks feed on at these locations. Only small variations of BMAA and DAB levels were observed and these were not correlated to any of the phytoplankton species reported. Therefore, extensive research should be performed on both origin and metabolism of BMAA in shellfish. The levels observed in this study are similar to those found in other studies in France or elsewhere. A previous study had related such levels to a cluster of Amyotrophic Lateral Sclerosis in the South of France; hence the widespread occurrence of BMAA in shellfish from all coasts in France found in this study suggests the need for further epidemiological and toxicological studies to establish the levels that are relevant for a link between the consumption of BMAA-containing foodstuffs and neurodegenerative diseases., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Pseudoalteromonas is widespread in various marine environments, and most strains can affect invertebrate larval settlement and metamorphosis by forming biofilms. However, the impact and the molecular basis of population diversification occurring in Pseudoalteromonas biofilms are poorly understood. Here, we show that morphological diversification is prevalent in Pseudoalteromonas species during biofilm formation. Two types of genetic variants, wrinkled (frequency of 12±5%) and translucent (frequency of 5±3%), were found in Pseudoalteromonas lipolytica biofilms. The inducing activities of biofilms formed by the two variants on larval settlement and metamorphosis of the mussel Mytilus coruscus were significantly decreased, suggesting strong antifouling activities. Using whole-genome re-sequencing combined with genetic manipulation, two genes were identified to be responsible for the morphology alternations. A nonsense mutation in AT00&#95;08765 led to a wrinkled morphology due to the overproduction of cellulose, whereas a point mutation in AT00&#95;17125 led to a translucent morphology via a reduction in capsular polysaccharide production. Taken together, the results suggest that the microbial behavior on larval settlement and metamorphosis in marine environment could be affected by the self-generated variants generated during the formation of marine biofilms, thereby rendering potential application in biocontrol of marine biofouling.

Benthic suspension feeding mussels are an important functional guild in coastal and estuarine ecosystems. To date we lack information on how various environmental gradients and biotic interactions separately and interactively shape the distribution patterns of mussels in non-tidal environments. Opposing to tidal environments, mussels inhabit solely subtidal zone in non-tidal waterbodies and, thereby, driving factors for mussel populations are expected to differ from the tidal areas. In the present study, we used the boosted regression tree modelling (BRT), an ensemble method for statistical techniques and machine learning, in order to explain the distribution and biomass of the suspension feeding mussel Mytilus trossulus in the non-tidal Baltic Sea. BRT models suggested that (1) distribution patterns of M. trossulus are largely driven by separate effects of direct environmental gradients and partly by interactive effects of resource gradients with direct environmental gradients. (2) Within its suitable habitat range, however, resource gradients had an important role in shaping the biomass distribution of M. trossulus. (3) Contrary to tidal areas, mussels were not competitively superior over macrophytes with patterns indicating either facilitative interactions between mussels and macrophytes or co-variance due to common stressor. To conclude, direct environmental gradients seem to define the distribution pattern of M. trossulus, and within the favourable distribution range, resource gradients in interaction with direct environmental gradients are expected to set the biomass level of mussels.

Predicting copper (Cu) toxicity in marine and estuarine environments is challenging because of the influence of anions on Cu speciation, competition between Cu(2+) and other cations at the biotic ligand and the effect of salinity on the physiology of the organism. In the present study the combined effect of salinity and dissolved organic carbon (DOC) on Cu toxicity to larvae of Mytilus galloprovincialis was assessed. Two statistical models were developed and used to elucidate the relationship between Cu toxicity, salinity, and DOC. All models based on dissolved Cu indicate a decrease in Cu toxicity with increasing DOC concentrations, which can partly be explained by complexation of Cu(2+) ions with DOC. These models also indicate an increase in Cu toxicity (modeled with dissolved Cu or Cu(2+) activity) with increasing salinity, suggesting a salinity-induced alteration in the physiology of the mussel larvae. When based on Cu body burdens, neither of the models indicates an effect of salinity or DOC. This shows that the Cu body burden is a more constant predictor of Cu toxicity, regardless of the water chemistry influencing Cu speciation or competition and possible physiological alterations or changes in Cu speciation or competition., (© 2015 SETAC.)

Most sedentary marine animals disperse from their place of origin during their initial life stages as larvae. The delivery of planktonic larvae back to coastal adult habitats after weeks or months of offshore development is commonly thought to be stochastic, resulting in large recruitment fluctuations and making predictive understanding of population dynamics difficult. Time series of invertebrate settlement on intertidal shores have been used to infer how various oceanographic processes deliver planktonic larvae ashore. However, the possibility that successful settlement may involve a series of different transport mechanisms, which are sequentially utilized by late-stage larvae, has received little attention. To address this, we monitored both the delivery of mussel and barnacle larvae to inner-shelf moorings positioned 200-1400 m from the shore, and larval settlement in the intertidal adult habitat, at two contrasting sites: a headland forming an upwelling center and a downstream bay. Model selection was employed to determine the most likely scenario(s) of larval onshore transport from four a priori transport mechanisms individually and in combination: (1) upwelling or relaxation/downwelling, (2) tidal motions, (3) diurnal sea breezes, and (4) surface waves. Mussel larvae were delivered to the inner shelf during upwelling in the bay, but during downwelling at the headland, and were further transported to the shore by surface waves at both locales. In contrast, the delivery of barnacle larvae to the inner shelf occurred during relaxation/downwelling events at both sites, and intertidal settlement coincided with spring tides, suggesting a role for internal tides in their onshore transport. Thus, sequential mechanisms appear to be utilized by larvae to get to the shore, involving interactions of regional-scale upwelling/downwelling processes and local-scale tidal and surface-wave processes, which differ among taxa and among sites with different topography. A bottleneck for larval delivery across the surf zone may be a result of out-of-phase steps in sequential transport mechanisms leaving larvae lost "in transit."

Nutritional quality parameters, microbiological and technological quality indicators (condition index, meat yield and water-holding capacity) of blue mussel, Mytilus galloprovincialis, reared in the North Adriatic Sea were characterised at monthly intervals over a 1 year period. Contents of protein (7.5-11.6 g/100 g), lipid (1.0-2.2 g/100 g) and ash (2.2-3.3 g/100 g) varied significantly accordingly to condition index (6-15%). n-3 PUFAs were the predominant fatty acids (38.7-45.9% of fatty acids) and docosahesaenoic and eicosapentaenoic acids were the most abundant (167 and 93.3 mg/100 g, respectively). Glycine, glutamic and aspartic acids accounted for 40% of total amino acids. All samples exhibited limited concentrations of Cr, Mn, Ni, Cu and Zn, as well as Na. M. galloprovincialis from the North Adriatic Sea showed the highest technological and nutritional quality, considering also the inter-annual variability, in late spring, which corresponds to the period immediately before gamete release., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A biometric, nutritional and sensory analysis of raw and cooked mussels comparing Mytilus sp. from the north-west coast of Portugal and Spain (Minho and Galicia, respectively) and the new offshore production site of Armona (Algarve, south Portugal) was carried out. In addition, multiple factorial analysis was performed to explore potential relationships between sensory attributes and nutritional content properties between the different mussels. Results showed that, at similar times of sale, biometrics of mussels from Armona and Vigo were similar and bigger than the remaining. Nonetheless, despite some similarities in proximate composition, mussels presented differences in lipid classes, fatty acid content and free amino acids profiles. These differences were not fully reflected in the sensory assessment by the panel, which were able to distinguish different production sites in raw specimens but displayed problems in discrimination these in cooked mussels. Some nutritional components were related to specific sensory sensations., (Copyright © 2014 Elsevier Ltd. All rights reserved.)