Molecular and behavioural responses of the mussel Mytilus edulis exposed to a marine heatwave.

Marine heatwaves (MHW) threaten marine organisms and tend to increase in frequency and intensity. We exposed the blue mussel Mytilus edulis to a MHW lasting 23 days, including two 10-d periods of thermal intensity increase of +5 °C (20 °C-25 °C) interspersed by 1 day back to 20 °C, followed by a 4-d recovery period. We investigated behaviour responses of mussels and gene expression changes relative to the circadian rhythm (Per), oxidative stress (SOD), cellular apoptosis (CASP3), energy production (ATPs), and general stress response (hsp70). Results showed that the MHW disturbed the valve activity of mussels. Particularly, mussels increased the number of valve micro-closures, showing a stressful state of organisms. Mussels also decreased Per, CASP3, ATPs, and Hsp70 gene expression. Some behavioural and molecular effects persisted after the MHW, suggesting a limited recovery capacity of individuals. This work highlighted the vulnerability of M. edulis to a realistic MHW.

Using valve gape analysis to compare sensitivity of native Mytilus edulis to invasive Magallana gigas when exposed to heavy metal contamination.

Coastal ecosystems are ecologically and economically important but are under increasing pressure from numerous anthropogenic sources of stress. Both heavy metal pollution and invasive species pose major environmental concerns that can have significant impacts on marine organisms. It is likely that many stresses will occur simultaneously, resulting in potential cumulative ecological effects. The aim of this study was to compare the relative resilience of an invasive oyster Magallana gigas and a native mussel Mytilus edulis to heavy metal pollution, utilising their valve gape response as an indicator. The gape activity of bivalves has been utilised to monitor a range of potential impacts, including for example oil spills, increased turbidity, eutrophication, heavy metal contamination etc. In this study, Hall effect sensors were used on both the native blue mussel (M. edulis) and the pacific oyster (M. gigas), invasive to Ireland. Mussels were shown to be more responsive to pollution events than oysters, where all heavy metals tested (copper, cadmium, zinc, lead) had an effect on transition frequency though significant differences were only observed for lead and cadmium (Control; > Copper, p = 0.0003; >lead, p = 0.0002; >Cadmium, p = 0.0001). Cadmium had an apparent effect on mussels with specimens from this treatment remaining closed for an average of 45.3% of the time. Similarly, significant effects on the duration of time mussels spent fully open was observed when treated with lead and cadmium (Control; > lead, p = 0.03, > cadmium, p = 0.02). In contrast, oysters displayed no significant difference for any treatment for number of gapes, or duration spent open or closed. Though there was an effect of both zinc and copper on the amount of time spent closed, with averages of 63.2 and 68.7% respectively. This indicates oysters may be potentially more resilient to such pollution events; further boosting their competitive advantage. Future mesocosm or field studies are required to quantify this relative resilience.

Size-dependent response of the mussel collective behaviour to plastic leachates and predator cues.

Both individual and collective anti-predator behaviours are essential for the survival of many species. This is particularly true for ecosystem engineers such as intertidal mussels, which through their collective behaviour create novel habitats for a range of organisms and biodiversity hotspots. However, contaminants may disrupt these behaviours and consequently indirectly affect exposure to predation risk at the population level. Among these, plastic litter is a major and ubiquitous contaminant of the marine environment. Here, we assessed the impact of microplastic (MP) leachates of the most produced plastic polymer, polypropylene (PlasticsEurope, 2022), at a high but locally relevant concentration (i.e. ca. 12 g L-1) on the collective behaviours and anti-predator responses of both small and large Mytilus edulis mussels. Indeed, in contrast to large mussels, small ones reacted to MP leachates, showing a taxis towards conspecifics and stronger aggregations. All mussels reacted to the chemical cues of the predatory crab, Hemigrapsus sanguineus, but with two different collective anti-predator behaviours. Small mussels only showed a taxis towards conspecifics when exposed to predator cues. This response was also found in large ones with a tendency to form more strongly bound aggregations and a considerable reduced activity, i.e. they significantly delayed their time to start to form aggregations and decreased their gross distance. These anti-predator behaviours were respectively inhibited and impaired in small and large mussels by MP leachates. The observed collective behavioural changes may reduce individual fitness by enhancing predation risk, particularly in small mussels that are the crab H. sanguineus's favourite preys. Given the key role of mussels as ecosystem engineers, our observations suggest that plastic pollution may have implication on M. edulis at the species level, but also enhancing a cascading effect towards a higher level of organisation such as population, community and ultimately structure and function of intertidal ecosystem.

Organic matter processing in a [simulated] offshore wind farm ecosystem in current and future climate and aquaculture scenarios.

The rapid development of blue economy and human use of offshore space triggered the concept of co-location of marine activities and is causing diverse local pressures on the environment. These pressures add to, and interact with, global challenges such as ocean acidification and warming. This study investigates the combined pressures of climate change and the planned co-location of offshore wind farm (OWF) and aquaculture zones on the carbon flow through epifaunal communities inhabiting wind turbines in the North Sea. A 13C-labelled phytoplankton pulse-chase experiment was performed in mesocosms (4 m3) holding undisturbed hard-substrate (HS) communities, natural sediment with infauna, and mobile invertebrate predators. Carbon assimilation was quantified under current and predicted future-climate conditions (+3 °C and -0.3 pH units), as well as a future-climate co-use scenario with blue mussel (Mytilus edulis) aquaculture. Climate change induced an increase in macrofaunal carbon assimilation as well as an organic enrichment of underlying sediments. Dynamic (non-)trophic links between M. edulis and other HS epifauna resulted in shifts among the species contributing most to the phytoplankton-derived carbon flow across climate scenarios. Increased inter- and intraspecific resource competition in the presence of M. edulis aquaculture prevented a large increase in the total assimilation of phytoplankton by HS fauna. Lower individual carbon assimilation rates by both mussels and other epifauna suggest that if filter capacity by HS epifauna would approach renewal by advection/mixing, M. edulis individuals would likely grow to a smaller-than-desired commercial size. In the same scenario, benthic organic carbon mineralisation was significantly boosted due to increased organic matter deposition by the aquaculture set-up. Combining these results with in situ OWF abundance data confirmed M. edulis as the most impactful OWF AHS species in terms of (total) carbon assimilation as well as the described stress responses due to climate change and the addition of bivalve aquaculture.

Microplastic leachates disrupt the chemotactic and chemokinetic behaviours of an ecosystem engineer (Mytilus edulis).

The massive contamination of the environment by plastics is an increasing global scientific and societal concern. Knowing whether and how these pollutants affect the behaviour of keystone species is essential to identify environmental risks effectively. Here, we focus on the effect of plastic leachates on the behavioural response of the common blue mussel Mytilus edulis, an ecosystem engineer responsible for the creation of biogenic structures that modify the environment and provide numerous ecosystem functions and services. Specifically, we assess the effect of virgin polypropylene beads on mussels' chemotactic (i.e. a directional movement in response to a chemical stimulus) and chemokinetic (i.e. a non-directional change in movement properties such as speed, distance travelled or turning frequency in response to a chemical stimulus) responses to different chemical cues (i.e. conspecifics, injured conspecifics and a predator, the crab Hemigrapsus sanguineus). In the presence of predator cues, individual mussels reduced both their gross distance and speed, changes interpreted here as an avoidance behaviour. When exposed to polypropylene leachates, mussels moved less compared to control conditions, regardless of the cues tested. Additionally, in presence of crab cues with plastic leachates, mussels significantly changed the direction of movement suggesting a leachate-induced loss of their negative chemotaxis response. Taken together, our results indicate that the behavioural response of M. edulis is cue-specific and that its anti-predator behaviour as well as its mobility are impaired when exposed to microplastic leachates, potentially affecting the functioning of the ecosystem that the species supports.

The survival and responses of blue mussel Mytilus edulis to 16-day sustained hypoxia stress.

The blue mussel Mytilus edulis, which is a worldwide commercial species distributed mainly from the intertidal zone to tens of meters deep, has been previously studied regarding its acute defense responses to air exposure and intermittent hypoxia. However, the effects of sustained hypoxia, such as caused by coastal eutrophication, remain to be explored. In the present study, the critical threshold of dissolved oxygen (DO) for experimental mussels exposed to 16 days of hypoxia was DO 0.7-0.8 mg L-1, below which survival dropped drastically from nearly 80% to <38%. When hypoxia was combined with DO fluctuations or with poor water quality, the threshold rose to an average of DO 1.0 mg L-1, which resulted in less than 80% survival. To find possible clues of physiological stress to account for mortalities, the metabolic rate and enzyme activities of Na+/K+ ATPase, superoxide dismutase, acid phosphatase, and alkaline phosphatase were further recorded.

Molecular Responses to Thermal and Osmotic Stress in Arctic Intertidal Mussels (Mytilus edulis): The Limits of Resilience.

Increases in Arctic temperatures have accelerated melting of the Greenland icesheet, exposing intertidal organisms, such as the blue mussel Mytilus edulis, to high air temperatures and low salinities in summer. However, the interaction of these combined stressors is poorly described at the transcriptional level. Comparing expression profiles of M. edulis from experimentally warmed (30 °C and 33 °C) animals kept at control (23‱) and low salinities (15‱) revealed a significant lack of enrichment for Gene Ontology terms (GO), indicating that similar processes were active under all conditions. However, there was a progressive increase in the abundance of upregulated genes as each stressor was applied, with synergistic increases at 33 °C and 15‱, suggesting combined stressors push the animal towards their tolerance thresholds. Further analyses comparing the effects of salinity alone (23‱, 15‱ and 5‱) showed high expression of stress and osmoregulatory marker genes at the lowest salinity, implying that the cell is carrying out intracellular osmoregulation to maintain the cytosol as hyperosmotic. Identification of aquaporins and vacuolar-type ATPase transcripts suggested the cell may use fluid-filled cavities to excrete excess intracellular water, as previously identified in embryonic freshwater mussels. These results indicate that M. edulis has considerable resilience to heat stress and highly efficient mechanisms to acclimatise to lowered salinity in a changing world.

Modeling the metabolic profile of Mytilus edulis reveals molecular signatures linked to gonadal development, sex and environmental site.

The monitoring of anthropogenic chemicals in the aquatic environment including their potential effects on aquatic organisms, is important for protecting life under water, a key sustainable development goal. In parallel with monitoring the concentrations of chemicals of concern, sentinel species are often used to investigate the biological effects of contaminants. Among these, bivalve molluscs such as mussels are filter-feeding and sessile, hence an excellent model system for measuring localized pollution. This study investigates the relationship between the metabolic state of the blue mussel (Mytilus edulis) and its physiology in different environments. We developed a computational model based on a reference site (relatively unpolluted) and integrated seasonal dynamics of metabolite relative concentrations with key physiological indicators and environmental parameters. The analysis of the model revealed that changes in metabolite levels during an annual cycle are influenced by water temperature and are linked to gonadal development. This work supports the importance of data-driven biology and its potential in environmental monitoring.

Integrated analysis of physiological, transcriptomics and metabolomics provides insights into detoxication disruption of PFOA exposure in Mytilus edulis.

Perfluorooctanoic acid (PFOA), a persistent environmental contaminant, resists environmental degradation and bioaccumulates in food chains. Lots of literatures have proved that PFOA exposure could disrupt detoxifying function in a variety of organisms, however, it still remained poorly known about this in mollusk. Here, we examined physiological, transcriptomic, and metabolomic responses to PFOA in Mytilus edulis, a model organism frequently used in studies of aquatic pollution. We aimed to characterize PFOA-induced stress responses and detoxification mechanisms. PFOA exposure significantly altered antioxidant enzyme activity levels and the abundances of lipid peroxidation products. In addition, transcriptomic analysis indicated that several genes associated with oxidative stress and detoxication were differentially expressed after PFOA exposure. In combination, transcriptomic and metabolomic analyses showed that PFOA exposure disturbed several metabolic processes in M. edulis, including the lipid metabolism, amino acid metabolism, and carbohydrate metabolism etc. Molecular examination and enzymes assay of PFOA-exposed M. edulis after a 7-day depuration period still did not recover to control levels. The Pathway enrichment analysis proved that several pathways related to detoxification, such as c-Jun N-terminal kinase (JNK) and p38-dependent mitogen-activated protein kinase (MAPK) pathway, Peroxisome proliferator-activated receptor γ (PPARγ) pathway etc, were obviously affected. The present work verifies firstly PFOA disruption to molluscan detoxification and identifies the key pathways to understand the molecular mechanisms thereof. This study provides new insights into the detoxication disruption invoked in response to PFOA exposure in M. edulis.

Sensitivity to oil dispersants: Effects on the valve movements of the blue mussel Mytilus edulis and the giant scallop Placopecten magellanicus, in sub-arctic conditions.

In response to accidental oil spills at sea, chemical oil dispersants are utilized to limit negative impacts on nearby littoral zones. However, current evidence suggests that such dispersants may be toxic to aquatic organisms. Blue mussels (Mytilus edulis) and giant scallops (Placopecten magellanicus) were exposed to different environmentally relevant concentrations of oil dispersant and their behavioural responses were closely monitored using high frequency (10Hz) valvometry. Behavioural valve responses included rapid closures when oil dispersant was added to the experimental tanks. At higher concentrations, the mussels remained closed throughout the exposure period. The giant scallop displayed escape behaviours (clapping) prior to mortality, suggesting toxicity of the oil dispersant. Relationships between different behavioural indicators and oil dispersant concentrations were observed for both species, but with different trends. While scallops demonstrated positive correlations between gaping behaviours and dispersant concentration, mussels exhibited a concentration threshold beyond which the gaping behaviour was characteristic of longer closure periods. This study highlights behavioural response differences consistent with bivalve-specific biological traits: the continuous valve closure of an intertidal species, M. edulis, firmly attached to the substrate, and the escapement behaviours of a semi-mobile subtidal species, P. magellanicus. From these observations, it appears that valvometry could be used as a tool for environmental assessments.

Warming and temperature variability determine the performance of two invertebrate predators.

In a warming ocean, temperature variability imposes intensified peak stress, but offers periods of stress release. While field observations on organismic responses to heatwaves are emerging, experimental evidence is rare and almost lacking for shorter-scale environmental variability. For two major invertebrate predators, we simulated sinusoidal temperature variability (±3 °C) around todays' warm summer temperatures and around a future warming scenario (+4 °C) over two months, based on high-resolution 15-year temperature data that allowed implementation of realistic seasonal temperature shifts peaking midpoint. Warming decreased sea stars' (Asterias rubens) energy uptake (Mytilus edulis consumption) and overall growth. Variability around the warming scenario imposed additional stress onto Asterias leading to an earlier collapse in feeding under sinusoidal fluctuations. High-peak temperatures prevented feeding, which was not compensated during phases of stress release (low-temperature peaks). In contrast, increased temperatures increased feeding on Mytilus but not growth rates of the recent invader Hemigrapsus takanoi, irrespective of the scale at which temperature variability was imposed. This study highlights species-specific impacts of warming and identifies temperature variability at the scale of days to weeks/months as important driver of thermal responses. When species' thermal limits are exceeded, temperature variability represents an additional source of stress as seen from future warming scenarios.

Effects of variable deoxygenation on trace element bioaccumulation and resulting metabolome profiles in the blue mussel (Mytilus edulis).

The dissolved oxygen concentration of the world's oceans has systematically declined by 2% over the past 50 years, and there has been a notable commensurate expansion of the global oxygen minimum zones (OMZs). Such wide-scale ocean deoxygenation affects the distribution of biological communities, impacts the physiology of organisms that may affect their capacity to absorb and process contaminants. Therefore, the bioaccumulation efficiencies of three contrasting radionuclides, 110mAg, 134Cs and 65Zn were investigated using controlled aquaria in the blue mussel Mytilus edulis under three contrasting dissolved oxygen regimes: normoxic; 7.14 mg L-1, reduced oxygen; 3.57 mg L-1 and hypoxic 1.78 mg L-1 conditions. Results indicated that hypoxic conditions diminished 110mAg uptake in the mussel, whereas depuration rates were not affected. Similarly, hypoxia appeared to cause a decrease in the 65Zn bioaccumulation rate, as evidenced by both weakened uptake and rapid elimination rates. Effects of hypoxia on the metabolome of mussels were also explored by untargeted Nuclear Magnetic Resonance (NMR) spectroscopic methods. The metabolic response was characterised by significantly greater abundance of several amino acids, amino sulfonic acids, dicarboxylic acids, carbohydrates and other metabolites in the lowest oxygen treatment, as compared to the higher oxygen treatments. Clearance rates significantly dropped in hypoxic conditions compared to normoxia. Results suggest that hypoxic conditions, and even partly moderate hypoxia, alter ventilation, an-aerobic, oxidative and osmoregulation metabolism of this mussel, which may further influence the trace element bioaccumulation capacity.

Effects of fullerene C60 in blue mussels: Role of mTOR in autophagy related cellular/tissue alterations.

The effects of C60 on mTOR (mechanistic Target of Rapamycin) activity in mussel digestive gland were investigated. mTOR is a kinase that senses physiological and environmental signals to control eukaryotic cell growth. mTOR is present in two complexes: the phosphorylated mTORC1 regulates cell growth by activating anabolic processes, and by inhibiting catabolic processes (i.e. autophagy); mTORC2 also modulates actin cytoskeleton organization. Mussels were exposed to C60 (0.01, 0.1 and 1 mg/L) for 72 h. Immunocytochemical analysis using a specific antibody revealed the cellular distribution of C60 in mussel digestive gland, already at the lowest concentration. In exposed mussels, the dephosphorylation of mTORC1 and mTORC2 may explain the C60 effects, i.e. the reduction of lysosomal membrane stability, the enhancement of LC3B protein, and the increase of lysosomal/cytoplasmic volume ratio; as well the cytoskeletal alterations. No oxidative stress was observed. Multivariate analysis was used to facilitate the interpretation of the biomarker data. Finally, a low density oligo-microarray was used to understand the cellular responses to fullerene. Transcriptomics identified a number of differentially expressed genes (DEGs) showing a maximum in animals exposed to 0.1 mg/L C60. The most affected processes are associated with energy metabolism, lysosomal activity and cytoskeleton organization. In this study, we report the first data on the subcellular distribution of C60 in mussel's cells; and on the involvement of mTOR inhibition in the alterations due to nanoparticle accumulation. Overall, mTOR deregulation, by affecting protein synthesis, energy metabolism and autophagy, may reduce the capacity of the organisms to effectively grow and reproduce.

Antioxidant defenses and metabolic responses of blue mussels (Mytilus edulis) exposed to various concentrations of erythromycin.

Erythromycin, one of the most widely used macrolide antibiotics, has been detected in various aquatic environments, so erythromycin ecotoxicity should deserve more attention. In this study, blue mussels (Mytilus edulis) were exposed to erythromycin to explore its potential physiological toxicity. After 2d acute and 7d sub-acute exposure to erythromycin, blue mussel glutathione S-transferase (GST) and catalase (CAT) activities were determined with microplate methods and metabolic responses were analyzed using 1H nuclear magnetic resonance (1H NMR). The results revealed that GST was approximately 1.6 times higher in exposed mussels at 200 mg/L and higher concentrations. CAT was about 1.9 times higher in exposed mussels at 200 mg/L, indicating that erythromycin exposure led that blue mussels enhanced antioxidant responses. Low doses of erythromycin exposure had a relatively small impact on the metabolism, while high doses of erythromycin exposure (200 and 400 mg/L) disturbed metabolic balance. With the increase of erythromycin concentrations, the individual metabolic differences within the same treatment groups also increased. The significant increase in alanine, glutamate, taurine, glycine and betaine were observed after acute and subacute exposure. Betaine played an important role in protecting antioxidant enzyme activities through adjusting osmotic pressure. The metabolomic results also showed the modes of erythromycin acted on the energy metabolism, osmoregulation, nerve activities and amino acid metabolism. This study highlighted how metabolomics can provide a comprehensive picture of metabolic responses, although significant antioxidant and metabolic responses were observed at high exposure concentrations.

Decreased thermal tolerance under recurrent heat stress conditions explains summer mass mortality of the blue mussel Mytilus edulis.

Extreme events such as heat waves have increased in frequency and duration over the last decades. Under future climate scenarios, these discrete climatic events are expected to become even more recurrent and severe. Heat waves are particularly important on rocky intertidal shores, one of the most thermally variable and stressful habitats on the planet. Intertidal mussels, such as the blue mussel Mytilus edulis, are ecosystem engineers of global ecological and economic importance, that occasionally suffer mass mortalities. This study investigates the potential causes and consequences of a mass mortality event of M. edulis that occurred along the French coast of the eastern English Channel in summer 2018. We used an integrative, climatological and ecophysiological methodology based on three complementary approaches. We first showed that the observed mass mortality (representing 49 to 59% of the annual commercial value of local recreational and professional fisheries combined) occurred under relatively moderate heat wave conditions. This result indicates that M. edulis body temperature is controlled by non-climatic heat sources instead of climatic heat sources, as previously reported for intertidal gastropods. Using biomimetic loggers (i.e. 'robomussels'), we identified four periods of 5 to 6 consecutive days when M. edulis body temperatures consistently reached more than 30 °C, and occasionally more than 35 °C and even more than 40 °C. We subsequently reproduced these body temperature patterns in the laboratory to infer M. edulis thermal tolerance under conditions of repeated heat stress. We found that thermal tolerance consistently decreased with the number of successive daily exposures. These results are discussed in the context of an era of global change where heat events are expected to increase in intensity and frequency, especially in the eastern English Channel where the low frequency of commercially exploitable mussels already questions both their ecological and commercial sustainability.

The influence of hydrodynamics and ecosystem engineers on eelgrass seed trapping.

Propagule dispersal is an integral part of the life cycle of seagrasses; important for colonising unvegetated areas and increasing their spatial distribution. However, to understand recruitment success, seed dispersal and survival in habitats of different complexity remains to be quantified. We tested the single and synergistic effects of three commonly distributed ecosystem engineers-eelgrass (Zostera marina), oysters (Magellana gigas) and blue mussels (Mytilus edulis)-on trapping of Z. marina seeds in a hydraulic flume under currents. Our results suggest that seed retention increases with habitat complexity and further reveal insights into the underlying mechanisms. In eelgrass canopy, trapping occurred mostly through direct blocking of a seed's pathway, while trapping in bivalve patches was mainly related to altered hydrodynamics in the lee side, i.e. behind each specimen. With increasing flow velocity (24-30 cm s-1 in eelgrass canopy, 18-30 cm s-1 in bivalve patches), modifications of the sediment surface through increased turbulence and erosive processes became more important and resulted in high seed trapping rates. Furthermore, we show that while monospecific patches of seagrass and bivalves had different trapping optima depending on flow velocities, intermixing resulted in consistently high trapping rates throughout the investigated hydrodynamic gradient. Our results highlight the importance of positive interactions among ecosystem engineers for seed retention and patch emergence in eelgrass.

Influence of blue mussel (Mytilus edulis) intake on fatty acid composition in erythrocytes and plasma phospholipids and serum metabolites in women with rheumatoid arthritis.

Intake of blue mussels decreased disease activity in women with rheumatoid arthritis (RA) in the randomized cross-over MIRA (Mussels, inflammation and RA) trial. This study investigates potential causes of the decreased disease activity by analysing fatty acid composition in erythrocytes and plasma phospholipids and serum metabolites in samples from the participants of the MIRA trial. Twenty-three women completed the randomized 2 × 11-week cross-over dietary intervention, exchanging one cooked meal per day, 5 days a week, with a meal including 75 g blue mussels or 75 g meat. Fatty acid composition in erythrocytes and plasma and 1H Nuclear Magnetic Resonance (1H NMR) metabolomics data were analysed with multivariate data analysis. Orthogonal Projections to Latent Structures with Discriminant Analysis (OPLS-DA) and OPLS with effect projections (OPLS-EP) were performed to compare the two diets. The fatty acid profile in erythrocytes was different after intake of blue mussels compared to the control diet, and all samples were correctly classified to either the blue mussel diet or control diet. Changes following blue mussel intake included significant increases in omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) at the group level but not for all individuals. The fatty acid profile in plasma phospholipids and 1H NMR serum metabolites did not differ significantly between the diets. To conclude, modelling fatty acids in erythrocytes may be a better biomarker for seafood intake than only EPA and DHA content. The change in fatty acid pattern in erythrocytes could be related to reduction in disease activity, although it cannot be excluded that other factors than omega-3 fatty acids potentiate the effect.

The effects of ZnO nanostructures of different morphology on bioenergetics and stress response biomarkers of the blue mussels Mytilus edulis.

Biofouling causes massive economical losses in the maritime sector creating an urgent need for effective and ecologically non-harmful antifouling materials. Zinc oxide (ZnO) nanorod coatings show promise as an antifouling material; however, the toxicity of ZnO nanorods to marine organisms is not known. We compared the toxicity of suspended ZnO nanorods (NR) with that of ZnO nanoparticles (NP) and ionic Zn2+ in a marine bivalve Mytilus edulis exposed for two weeks to 10 or 100 µg Zn L-1 of ZnO NPs, NRs or Zn2+, or to immobilized NRs. The multi-biomarker assessment included bioenergetics markers (tissue energy reserves, activity of mitochondrial electron transport system and autophagic enzymes), expression of apoptotic and inflammatory genes, and general stress biomarkers (oxidative lesions, lysosomal membrane stability and metallothionein expression). Exposure to ZnO NPs, NRs and Zn2+ caused accumulation of oxidative lesions in proteins and lipids, stimulated autophagy, and led to lysosomal membrane destabilization indicating toxicity. However, these responses were not specific for the form of Zn (NPs, NR or Zn2+) and showed no monotonous increase with increasing Zn concentrations in the experimental exposures. No major disturbance of the energy status was found in the mussels exposed to ZnO NPs, NRs, or Zn2+. Exposure to ZnO NPs and NRs led to a strong induction of apoptosis- and inflammation-related genes, which was not seen in Zn2+ exposures. Based on the integrated biomarker response, the overall toxicity as well as the pro-apoptotic and pro-inflammatory action was stronger in ZnO NPs compared with the NRs. Given the stability of ZnO NR coatings and the relatively low toxicity of suspended ZnO NR, ZnO NR coating might be considered a promising low-toxicity material for antifouling paints.

Hypoxia aggravates the effects of ocean acidification on the physiological energetics of the blue mussel Mytilus edulis.

Apart from ocean acidification, hypoxia is another stressor to marine organisms, especially those in coastal waters. Their interactive effects of elevated CO2 and hypoxia on the physiological energetics in mussel Mytilus edulis were evaluated. Mussels were exposed to three pH levels (8.1, 7.7, 7.3) at two dissolved oxygen levels (6 and 2 mg L-1) and clearance rate, absorption efficiency, respiration rate, excretion rate, scope for growth and O: N ratio were measured during a14-day exposure. After exposure, all parameters (except excretion rate) were significantly reduced under low pH and hypoxic conditions, whereas excretion rate was significantly increased. Additive effects of low pH and hypoxia were evident for all parameters and low pH appeared to elicit a stronger effect than hypoxia (2.0 mg L-1). Overall, hypoxia can aggravate the effects of acidification on the physiological energetics of mussels, and their populations may be diminished by these stressors.

Potential nitrous oxide production by marine shellfish in response to warming and nutrient enrichment.

Bivalves facilitate microbial nitrogen cycling, which can produce nitrous oxide (N2O), a potent greenhouse gas. Potential N2O production by three marine bivalves (Mytilus edulis, Mercenaria mercenaria and Crassostrea virginica) was measured in the laboratory including responses to nitrogen (N) loading and/or warming over short-terms (up to 14 or 28 days). N additions (targeting 100 µM-N ammonium nitrate) or warming (22 °C) individually and in combination were applied with experimental controls (20 µM-N, 19 °C). N2O production rates were higher with N additions for all species, but warming lacked significant direct effects. Ammonium and nitrate concentrations varied but were consistent with nitrification as a potential N2O source for all bivalves. Highest N2O emissions (7.5 nmol N2O g-1 h-1) were from M. edulis under hypoxic conditions coincident with a drop in pH. Macro-epifauna on M. edulis did not significantly alter N2O production. Thus, under short-term hypoxic conditions, micro-organisms in M. edulis guts may be a particularly significant source of N2O.