The effect of different types of microplastic and acute cadmium exposure on the Mytilus galloprovincialis (Lamarck, 1819).

Microplastic (MP) pollution is a pressing issue for both environmental health and the safety of human food sources. This study provides a comprehensive analysis of the effects of MPs on Mediterranean mussels (Mytilus galloprovincialis, Lamarck 1819), focusing on the food safety risks associated with MP and cadmium (Cd) exposure in these organisms intended for consumption. The retention of different polymer types of MPs in mussels was specifically evaluated, and the influence of Cd on MP retention across these polymers was investigated. Mussels were exposed to polystyrene (PS), polypropylene (PP), and polyethylene terephthalate (PET) MPs individually and in combination with the toxic metal Cd for a duration of 7 days. Antioxidant enzymes, oxidative stress parameters, and digestive system enzyme activities, selected as biomarkers for Cd and MPs pollution, were assessed. Furthermore, human consumption risk evaluations and limits regarding mussel intake were analysed in terms of food safety. The results suggest that exposure to Cd, MPs, or their combination induces oxidative stress, tissue damage, and neurotoxicity. Alterations in digestive enzyme activities could impact the mussels' energy acquisition from food and their capacity to conserve energy reserves. The estimated daily intake (EDI), provisional tolerable weekly intake (PTWI), target hazard quotient (THQ), and target cancer risk (TCR) levels for all groups surpassed established limits, implying a significant health risk for humans consuming these products. These results underscore the potential health risks for humans associated with consuming mussels exposed to Cd and/or MPs and provide valuable data for monitoring pollution levels and ecological risks in aquatic organisms. Additionally, our findings reveal that the retention of Cd in mussel tissues varies significantly after exposure, with combinations of PET and Cd showing lower levels of Cd accumulation compared to other groups, suggesting a differential interaction that influences Cd retention.

Using marine mussels to assess the potential ecotoxicological effects of two different commercial microplastics.

Microplastics (MPs) in the aquatic environment pose a serious threat to biota, by being confounded with food. These effects occur in mussels which are filter-feeding organisms. Mussels from the genus Mytilus sp. were used to evaluate the ecotoxicological effects of two MPs, polypropylene (PP) and polyethylene terephthalate (PET), after 4 and 28-days. Measured individual endpoints were condition index and feeding rate; and sub-individual parameters, metabolism of phase I (CYP1A1, CYP1A2 and CYP3A4) and II (glutathione S-transferases - GSTs), and antioxidant defense (catalase - CAT). MPs decreased both condition index (CI) and feeding rate (FR). No alterations occurred in metabolic enzymes, suggesting that these MPs are not metabolized by these pathways. Furthermore, lack of alterations in GSTs and CAT activities suggests the absence of conjugation and oxidative stress. Overall, biochemical markers were not responsive, but non-enzymatic responses showed deleterious effects caused by these MPs, which may be of high ecological importance.

Bioaccumulation and biological effects of hydrogenated cement particles in the marine bivalve, Mytilusgalloprovincialis.

The decommissioning and normal functioning of nuclear facilities can result in the production and release of airborne particles in the environment. Aquatic biota are expected to be exposed to these particles considering that nuclear facilities are often located near water bodies. Aerosols, such as cement dust, can interact with radionuclides as well as with heavy metals, and therefore elicit not only radiological impacts but also chemical toxicity. In the present study, we aimed to determine the effects of hydrogenated cement particles (HCPs) as a first step before evaluating any radiotoxicity of tritiated cement particles in the marine mussels, Mytilus galloprovincialis. Responses at different levels of biological organisation were assessed, including clearance rate (CR), tissue specific accumulation, DNA damage and transcriptional expression of key stress related genes. Acute (5 h) and medium-term, chronic (11 d) exposures to 1000 µg L-1 HCPs showed that bioaccumulation, assessed using Cu as a proxy and determined by inductively coupled plasma mass spectrometry, was time and tissue dependent. The highest levels of Cu were found in the digestive gland (DG) after 11 d. HCP exposure caused changes in the expression of oxidative and other stress-related genes, including mt20 in DG and gst and sod in the gill after 5 h exposure, while an overexpression of hsp70 in the gill was observed after 11 d. Genotoxic effects in haemocytes were observed after 11 d of HCP exposure. Multivariate analysis indicated that oxidative stress is the most probable factor contributing to overall physiological dysfunction. Our results provide a baseline to perform further studies employing tritiated cement particles. Specifically, future work should focus on the DG since only this tissue showed significant bioaccumulation when compared to the negative control.

Exposure to environmental pharmaceuticals affects the macromolecular composition of mussels digestive glands.

Human pharmaceuticals represent a major challenge in natural environment. A better knowledge on their mechanisms of action and adverse effects on cellular pathways is fundamental to predict long-term consequences for marine wildlife. The FTIRI Imaging (FTIRI) spectroscopy represents a vibrational technique allowing to map specific areas of non-homogeneous biological samples, providing a unique biochemical and ultrastructural fingerprint of the tissue. In this study, FTIRI technique has been applied, for the first time, to characterize (i) the chemical building blocks of digestive glands of Mytilus galloprovincialis, (ii) alterations and (iii) resilience of macromolecular composition, after a 14-days exposure to 0.5 µg/L of carbamazepine (CBZ), valsartan (VAL) and their mixture, followed by a 14-days recovery period. Spectral features of mussels digestive glands provided insights on composition and topographical distribution of main groups of biological macromolecules, such as proteins, lipids, and glycosylated compounds. Pharmaceuticals caused an increase in the total amount of protein and a significant decrease of lipids levels. Changes in macromolecular features reflected the modulation of specific molecular and biochemical pathways thus supporting our knowledge on mechanisms of action of such emerging pollutants. Overall, the applied approach could represent an added value within integrated strategies for the effects-based evaluation of environmental contaminants.

Unraveling the interplay between environmental microplastics and salinity stress on Mytilus galloprovincialis larval development: A holistic exploration.

The rise of plastic production has triggered a surge in plastic waste, overwhelming marine ecosystems with microplastics. The effects of climate change, notably changing salinity, have shaped the dynamics of coastal lagoons. Thus, understanding the combined impact of these phenomena on marine organisms becomes increasingly crucial. To address these knowledge gaps, we investigated for the first time the interactive effects of environmental microplastics (EMPs) and increased salinity on the early development of Mytilus galloprovincialis larvae. Morphological assessments using the larval embryotoxicity test revealed larval anomalies and developmental arrests induced by EMPs and increased salinity. Transcriptomic analyses targeting 12 genes involved in oxidative stress, apoptosis, DNA repair, shell formation, and stress proteins were conducted on D-larvae uncovered the potential effects of EMPs on shell biomineralization, highlighting the role of Histidine Rich Glycoproteine (HRG) and tubulin as crucial adaptive mechanisms in Mytilus sp. in response to environmental shifts. Furthermore, we explored oxidative stress and neurotoxicity using biochemical assays. Our findings revealed a potential interaction between EMPs and increased salinity, impacting multiple physiological processes in mussel larvae. Our data contribute to understanding the cumulative effects of emerging anthropogenic pollutants and environmental stressors, emphasizing the need for a holistic approach to assessing their impact on marine ecosystems.

Can exposure to Gymnodinium catenatum toxic blooms influence the impacts induced by Neodymium in Mytilus galloprovincialis mussels? What doesn't kill can make them stronger?

The presence in marine shellfish of toxins and pollutants like rare earth elements (REEs) poses a major threat to human well-being, coastal ecosystems, and marine life. Among the REEs, neodymium (Nd) stands out as a widely utilized element and is projected to be among the top five critical elements by 2025. Gymnodinum catenatum is a phytoplankton species commonly associated with the contamination of bivalves with paralytic shellfish toxins. This study evaluated the biological effects of Nd on the mussel species Mytilus galloprovincialis when exposed to G. catenatum cells for fourteen days, followed by a recovery period in uncontaminated seawater for another fourteen days. After co-exposure, mussels showed similar toxin accumulation in the Nd and G. catenatum treatment in comparison with the G. catenatum treatment alone. Increased metabolism and enzymatic defenses were observed in organisms exposed to G. catenatum cells, while Nd inhibited enzyme activity and caused cellular damage. Overall, this study revealed that the combined presence of G. catenatum cells and Nd, produced positive synergistic effects on M. galloprovincialis biochemical responses compared to G. catenatum alone, indicating that organisms' performance may be significantly modulated by the presence of multiple co-occurring stressors, such those related to chemical pollution and harmful algal blooms. ENVIRONMENTAL IMPLICATIONS: Neodymium (Nd) is widely used in green technologies like wind turbines, and this element's potential threats to aquatic environments are almost unknown, especially when co-occurring with other environmental factors such as blooms of toxic algae. This study revealed the cellular impacts induced by Nd in the bioindicator species Mytilus galloprovincialis but further demonstrated that the combination of both stressors can generate a positive defense response in mussels. The present findings also demonstrated that the impacts caused by Nd lasted even after a recovery period while a previous exposure to the toxins generated a faster biochemical improvement by the mussels.

Nano-titanium dioxide exacerbates the harmful effects of perfluorooctanoic acid on the health of mussels.

Exposing marine organisms to contemporary contaminants, such as perfluorooctanoic acid (PFOA) and nano-titanium dioxide (nano-TiO2), can induce multifaceted physiological consequences. Our investigation centered on the responses of the mussel, Mytilus coruscus, to these agents. We discerned pronounced disruptions in gill filament connections, pivotal structures for aquatic respiration, suggesting compromised oxygen uptake capabilities. Concurrently, the respiratory rate exhibited a marked decline, indicating a respiratory distress. Furthermore, the mussels' clearance rate, a metric of their filtration efficacy, diminished, suggesting the potential for bioaccumulation of deleterious substances. Notably, the co-exposure of PFOA and nano-TiO2 exhibits interactive effects on the physiological performance of the mussels. The mussels' digestive performance waned in the face of heightened PFOA and nano-TiO2 concentrations, possibly hampering nutrient assimilation and energy accrual. This was mirrored in the noticeable contraction of their energy budget, suggesting long-term growth repercussions. Additionally, the dysregulation of the gut microbiota and the reduction in its diversity further confirm alterations in intestinal homeostasis, subsequently impacting its physiological functions and health. Collectively, these findings underscore the perils posed by escalated PFOA and nano-TiO2 levels to marine mussels, accentuating the need for a deeper understanding of nanoparticle-pollutant synergies in marine ecosystems.

Preliminary hazard assessment of a new nature-inspired antifouling (NIAF) agent.

A recently synthesized aminated 3,4-dioxygenated xanthone (Xantifoul2) was found to have promising antifouling (AF) effects against the settlement of the macrofouler Mytilus galloprovincialis larvae. Preliminary assessment indicated that Xantifoul2 has reduced ecotoxicological impacts: e.g., being non-toxic to the marine crustacea Artemia salina (<10 % mortality at 50 µM) and showing low bioconcentration factor in marine organisms. In order to meet the EU Biocidal Product Regulation, a preliminary hazard assessment of this new nature-inspired antifouling (NIAF) agent was conducted in this work. Xantifoul2 did not affect the swimming ability of the planktonic crustacean Daphnia magna, the growth of the diatom Phaeodactylum tricornutum, and the cellular respiration of luminescent Gram-negative bacteria Vibrio fischeri, supporting the low toxicity towards several non-target marine species. Regarding human cytotoxicity, Xantifoul2 did not affect the cell viability of retinal human cells (hTERT-RPE-1) and lipidomic studies revealed depletion of lipids involved in cell death, membrane modeling, lipid storage, and oxidative stress only at a high concentration (10 µM). Accelerated degradation studies in water were conducted under simulated sunlight to allow the understanding of putative transformation products (TPs) that could be generated in the aquatic ecosystems. Both Xantifoul2 and photolytic-treated Xantifoul2 in the aqueous matrix were therefore evaluated on several nuclear receptors (NRs). The results of this preliminary hazard assessment of Xantifoul2, combined with the high degradation rates in water, provide strong evidence of the safety of this AF agent under the evaluated conditions, and provide the support for future validation studies before this compound can be introduced in the market.

The role of the macroalgae Ulva lactuca on the cellular effects of neodymium and mercury in the mussel Mytilus galloprovincialis.

Rare earth elements (REEs) are increasingly being studied mainly due to their economic importance and wide range of applications, but also for their rising environmental concentrations and potential environmental and ecotoxicological impacts. Among REEs, neodymium (Nd) is widely used in lasers, glass additives, and magnets. Currently, NdFeB-based permanent magnets are the most significant components of electronic devices and Nd is used because of its magnetic properties. In addition to REEs, part of the environmental pollution related to electrical and electronic equipment, fluorescent lamps and batteries also comes from mercury (Hg). Since both elements persist in ecosystems and are continuously accumulated by marine organisms, a promising approach for water decontamination has emerged. Through a process known as sorption, live marine macroalgae can be used, especially Ulva lactuca, to accumulate potential toxic elements from the water. Therefore, the present study aimed to evaluate the cellular toxicity of Nd and Hg in Mytilus galloprovincialis, comparing the biochemical effects induced by these elements in the presence or absence of the macroalgae U. lactuca. The results confirmed that Hg was more toxic to mussels than Nd, but also showed the good capability of U. lactuca in preventing the onset of cellular disturbance and homeostasis disruption in M. galloprovincialis by reducing bioavailable Hg levels. Overall, the biochemical parameters evaluated related to metabolism, antioxidant and biotransformation defences, redox balance, and cellular damage, showed that algae could prevent biological effects in mussels exposed to Hg compared to those exposed to Nd. This study contributes to the advancement of knowledge in this field, namely the understanding of the impacts of different elements on bivalves and the crucial role of algae in the protection of other aquatic organisms.

Physiological response of mussel to rayon microfibers and PCB's exposure: Overlooked semi-synthetic micropollutant?

Rayon microfibers, micro-sized semi-synthetic polymers derived from cellulose, have been frequently detected and reported as "micropollutants" in marine environments. However, there has been limited research on their ecotoxicity and combined effects with persistent organic pollutants (POPs). To address these knowledge gaps, thick-shell mussels (Mytilus coruscus) were exposed to rayon microfibers at 1000 pieces/L, along with polychlorinated biphenyls (PCBs) at 100 and 1000 ng/L for 14 days, followed by a 7-day recovery period. We found that rayon microfibers at the environmentally relevant concentration exacerbated the irreversible effects of PCBs on the immune and digestive systems of mussels, indicating chronic and sublethal impacts. Furthermore, the results of 16 s rRNA sequencing demonstrated significant effects on the community structure, species richness, and diversity of the mussels' intestinal microbiota. The branching map analysis identified the responsive bacteria to rayon microfibers and PCBs belonging to the Proteobacteria, Actinobacteriota, and Bacteroidota phyla. Despite not being considered a conventional plastic, the extensive and increasing use of rayon fibers, their direct toxicological effects, and their interaction with POPs highlight the need for urgent attention, investigation, and regulation to address their contribution to "micropollution".

Mixtures of environmental pharmaceuticals in marine organisms: Mechanistic evidence of carbamazepine and valsartan effects on Mytilus galloprovincialis.

Unravelling the adverse outcomes of pharmaceuticals mixture represents a research priority to characterize the risk for marine ecosystems. The present study investigated, for the first time, the interactions between two of the most largely detected pharmaceuticals in marine species: carbamazepine (CBZ) and valsartan (VAL), elucidating mechanisms that can modulate bioaccumulation, excretion and the onset of toxicity. Mytilus galloprovincialis were exposed to environmental levels of CBZ and VAL dosed alone or in combination: measurement of drug bioaccumulation was integrated with changes in the whole transcriptome and responsiveness of various biochemical and cellular biomarkers. Interactive and competing mechanisms between tested drugs were revealed by the much higher CBZ accumulation in mussels exposed to this compound alone, while an opposite trend was observed for VAL. A complex network of responses was observed as variations of gene expression, functional effects on neurotransmission, cell cycle, immune responses and redox homeostasis. The elaboration of results through a quantitative Weight of Evidence model summarized a greater biological reactivity of CBZ compared to VAL and antagonistic interactions between these compounds, resulting in a reduced effect of the antiepileptic when combined with valsartan. Overall, new perspectives are highlighted for a more comprehensive risk assessment of environmental mixtures of pharmaceuticals.

Toxicity of two drugs towards the marine filter feeder Mytilus spp, using biochemical and shell integrity parameters.

The increasing presence of anthropogenic contaminants in the environment may constitute a challenge to non-target biota, considering that most contaminants can exert deleterious effects. Salicylic acid (SA) is a non-steroid anti-inflammatory drug (NSAID) which exerts its activity by inhibiting the enzyme cyclooxygenase (COX). Another class of drugs is that of the diuretics, in which acetazolamide (ACZ) is included. This pharmaceutical acts by inhibiting carbonic anhydrase (CA), a key enzyme in acid-base homeostasis, regulation of pH, being also responsible for the bio-availability of Ca2+ for shell biomineralization processes. In this work, we evaluated the chronic (28-day) ecotoxicological effects resulting from the exposures to SA and ACZ (alone, and in combination) on individuals of the marine mussel species Mytillus spp., using enzymatic (catalase (CAT), glutathione S-transferases (GSTs), COX and CA), non-enzymatic (lipid peroxidation, TBARS levels) and morphological and physiological (shell hardness, shell index and feeding behaviour) biomarkers. Exposure to ACZ and SA did not cause significant alterations in CAT and GSTs activities, and in TBARS levels. In terms of CA, this enzyme was inhibited by the highest concentration of ACZ in gills of exposed animals, but no effects occurred in the mantle tissue. The activity of COX was not altered after exposure to the single chemicals. However, animals exposed to the mixture of ACZ and SA evidenced a significant inhibition of COX activity. Morphological and physiological processes (namely, feeding, shell index, and shell hardness) were not affected by the here tested pharmaceutical drugs. Considering the general absence of adverse effects, further studies are needed to fully evaluate the effects of these pharmaceutical drugs on alternative biochemical and physiological pathways.

Larval metamorphosis is inhibited by methimazole and propylthiouracil that reveals possible hormonal action in the mussel Mytilus coruscus.

Larval metamorphosis in bivalves is a key event for the larva-to-juvenile transformation. Previously we have identified a thyroid hormone receptor (TR) gene that is crucial for larvae to acquire "competence" for the metamorphic transition in the mussel Mytilus courscus (Mc). The mechanisms of thyroid signaling in bivalves are still largely unknown. In the present study, we molecularly characterized the full-length of two iodothyronine deiodinase genes (McDx and McDy). Phylogenetic analysis revealed that deiodinases of molluscs (McDy, CgDx and CgDy) and vertebrates (D2 and D3) shared a node representing an immediate common ancestor, which resembled vertebrates D1 and might suggest that McDy acquired specialized function from vertebrates D1. Anti-thyroid compounds, methimazole (MMI) and propylthiouracil (PTU), were used to investigate their effects on larval metamorphosis and juvenile development in M. coruscus. Both MMI and PTU significantly reduced larval metamorphosis in response to the metamorphosis inducer epinephrine. MMI led to shell growth retardation in a concentration-dependent manner in juveniles of M. coruscus after 4 weeks of exposure, whereas PTU had no effect on juvenile growth. It is hypothesized that exposure to MMI and PTU reduced the ability of pediveliger larvae for the metamorphic transition to respond to the inducer. The effect of MMI and PTU on larval metamorphosis and development is most likely through a hormonal signal in the mussel M. coruscus, with the implications for exploring the origins and evolution of metamorphosis.

Expression Analyses of Genes Related to Multixenobiotic Resistance in Mytilus galloprovincialis after Exposure to Okadaic Acid-Producing Dinophysis acuminata.

The mussel Mytilus galloprovincialis is one of the most important aquaculture species in Europe. Its main production problem is the accumulation of toxins during coastal blooms, which prevents mussel commercialization. P-glycoprotein (ABCB1/MDR1/P-gp) is part of the multixenobiotic resistance system in aquatic organisms, and okadaic acid, the main DSP toxin, is probably a substrate of the P-gp-mediated efflux. In this study, the presence and possible role of P-gp in the okadaic acid detoxification process was studied in M. galloprovincialis. We identified, cloned, and characterized two complete cDNAs of mdr1 and mdr2 genes. MgMDR1 and MgMDR2 predicted proteins had the structure organization of ABCB full transporters, and were identified as P-gp/MDR/ABCB proteins. Furthermore, the expression of mdr genes was monitored in gills, digestive gland, and mantle during a cycle of accumulation-elimination of okadaic acid. Mdr1 significantly increased its expression in the digestive gland and gills, supporting the idea of an important role of the MDR1 protein in okadaic acid efflux out of cells in these tissues. The expression of M. galloprovincialismrp2, a multidrug associated protein (MRP/ABCC), was also monitored. As in the case of mdr1, there was a significant induction in the expression of mrp2 in the digestive gland, as the content of okadaic acid increased. Thus, P-gp and MRP might constitute a functional defense network against xenobiotics, and might be involved in the resistance mechanisms to DSP toxins.

Multi-biomarker approach in Mytilus galloprovincialis and Ruditapes decussatus as a predictor of pelago-benthic responses after exposure to Benzo[a]Pyrene.

This study evaluated the biomarker responses indicative of exposure to Benzo[a] Pyrene (B[a]P) in Mytilus galloprovincialis and Ruditapes decussatus. A significant increase of the total oxyradical scavenging capacity (TOSC) was observed after seven days of exposure to two concentrations of B[a]P (100 and 300 µg.L-1), in the digestive gland with the lowest concentration tested. The TOSC in the gills increased notably only after the exposure to 300 µg.L-1 of B[a]P. Interestingly, the superoxide dismutase (SOD) and catalase (CAT) activities in gills and digestive gland on one hand and glutathione S-transferase (GST) in gills in the other, were positively correlated with the concentration of B[a]P with a significant induction noticed at the highest concentration. In contrast, a significant increase of the GST activity was observed in the digestive gland following the exposure of bivalves to 100 µg.L-1. In pelagic (M. galloprovincialis) or benthic (R. decussatus) bivalves, the AChE activity decreased discernibly in digestive glands and gills with the increase of B[a]P concentrations as evidence of neurotoxic effects. In clams, the exposure to B[a]P was followed by a significant increase of Malondialdehyde level (MDA) in gills and digestive gland, this does not occur in gills of Mytilus galloprovincialis at the concentration of 100 µg.L-1. Overall, the results found seems to indicate that the mussel was more suitable as a predictor tool of toxicity of B[a]P.

The cadmium toxicity in gills of Mytilus coruscus was accentuated by benzo(a)pyrene of higher dose but not lower dose.

In natural environment, the existence of interactions of toxic mixtures could induce diverse biochemical pathways and consequently exert different toxicological responses in aquatic organisms. However, little information is available on the effects of combined xenobiotics on lower aquatic invertebrates. Here, we assessed the effects of cadmium (Cd, 0.31 mg/L) as well as the mixture of Cd (0.31 mg/L) and benzo(a)pyrene (Bap, 5 or 50 µg/L) on bioaccumulation, antioxidant, lipid peroxidation (LPO) and metallothionein (MT) responses in gills of thick shell mussel Mytilus coruscus. Upon exposed to single Cd, the metal bioaccumulation, antioxidant enzymes activities, LPO and MT level significantly increased in the gills, suggesting an apparent toxicity to mussels. The interaction of Cd + 5 µg/L Bap did not significantly alter these endpoints compared to single Cd. However, once the dose of Bap elevated to 50 µg/L, the induction of bioaccumulation, antioxidant system and LPO was even more pronounced while the induction of MT was remarkably inhibited, implying an accentuated toxicity. Collectively, the current results demonstrated that 0.31 mg/L Cd exposure resulted in severe toxicity to mussels despite of the induction of MT system to alleviate the metal toxicity. Once the Cd exposure combined with Bap, the lower dose of Bap could not change the Cd toxicity while the higher dose of Bap accentuated the toxicity by inhibiting metallothionein synthesis. These findings might provide some useful clues for elucidation the mechanism of the interaction of combined xenobiotics in molluscs.

The influence of salinity on sodium lauryl sulfate toxicity in Mytilus galloprovincialis.

The influence of salinity on the effects of sodium lauryl sulfate (SLS) was evaluated using the Mediterranean mussel Mytilus galloprovincialis, exposed for 28 days to SLS (control-0.0 and 4.0 mg/L) under three salinity levels (Control-30, 25 and 35). The effects were monitored using biomarkers related to metabolism and energy reserves, defence mechanisms (antioxidant and biotransformation enzymes) and cellular damage. The results revealed that non-contaminated mussels tended to maintain their metabolic capacity regardless of salinity, without activation of antioxidant defence strategies. On the contrary, although contaminated mussels presented decreased metabolic capacity at salinities 25 and 35, they were able to activate their antioxidant mechanisms, preventing cellular damage. Overall, the present findings indicate that SLS, especially under stressful salinity levels, might potentially jeopardize population survival and reproduction success since reduced metabolism and alterations on mussels' antioxidant mechanisms will impair their biochemical and, consequently, physiological performance.

The Endocannabinoid System in the Mediterranean Mussel Mytilus galloprovincialis: Possible Mediators of the Immune Activity?

Anandamide (AEA) is one of the best characterized members of the endocannabinoid family and its involvement in many pathophysiological processes has been well documented in vertebrates and invertebrates. Here, we report the biochemical and functional characterization of key elements of the endocannabinoid system in hemocytes isolated from the Mediterranean mussel Mytilus galloprovincialis. We also show the effects of exogenous AEA, as well as of capsaicin, on the cell ability to migrate and to activate the respiratory burst, upon in vitro stimulation of phagocytosis. Interestingly, our findings show that both AEA and capsaicin suppress the hemocyte response and that the use of selective antagonists of CB2 and TRPV1 receptors revert their inhibitory effects. Overall, present data support previous evidence on the presence of endocannabinoid signaling in mollusks and advance our knowledge about the evolutionary origins of this endogenous system and its role in the innate response of mollusks.

How Ulva lactuca can influence the impacts induced by the rare earth element Gadolinium in Mytilus galloprovincialis? The role of macroalgae in water safety towards marine wildlife.

Rare earth elements (REEs) are gaining growing attention in environmental and ecotoxicological studies due to their economic relevance, wide range of applications and increasing environmental concentrations. Among REEs, special consideration should be given to Gadolinium (Gd), whose wide exploitation as a magnetic resonance imaging (MRI) contrast agent is enhancing the risk of its occurrence in aquatic environments and impacts on aquatic organisms. A promising approach for water decontamination from REEs is sorption, namely through the use of macroalgae and in particular Ulva lactuca that already proved to be an efficient biosorbent for several chemical elements. Therefore, the present study aimed to evaluate the toxicity of Gd, comparing the biochemical effects induced by this element in the presence or absence of algae. Using the bivalve species Mytilus galloprovincialis, Gd toxicity was evaluated by assessing changes on mussels' metabolic capacity and oxidative status. Results clearly showed the toxicity of Gd but further revealed the capacity of U. lactuca to prevent injuries to M. galloprovincialis, mainly reducing the levels of Gd in water and thus the bioaccumulation and toxicity of this element by the mussels. The results will advance the state of the art not only regarding the effects of REEs but also with regard to the role of algae in accumulation of metals and protection of aquatic organisms, generating new insights on water safety towards aquatic wildlife and highlighting the possibility for resources recovery.

Spermatozoa Transcriptional Response and Alterations in PL Proteins Properties after Exposure of Mytilus galloprovincialis to Mercury.

Mercury (Hg) is an environmental pollutant that impacts human and ecosystem health. In our previous works, we reported alterations in the properties of Mytilus galloprovincialis protamine-like (PL) proteins after 24 h of exposure to subtoxic doses of toxic metals such as copper and cadmium. The present work aims to assess the effects of 24 h of exposure to 1, 10, and 100 pM HgCl2 on spermatozoa and PL proteins of Mytilus galloprovincialis. Inductively coupled plasma-mass spectrometry indicated accumulation of this metal in the gonads of exposed mussels. Further, RT-qPCR analyses showed altered expression levels of spermatozoa mt10 and hsp70 genes. In Mytilus galloprovincialis, PL proteins represent the major basic component of sperm chromatin. These proteins, following exposure of mussels to HgCl2, appeared, by SDS-PAGE, partly as aggregates and showed a decreased DNA-binding capacity that rendered them unable to prevent DNA damage, in the presence of CuCl2 and H2O2. These results demonstrate that even these doses of HgCl2 exposure could affect the properties of PL proteins and result in adverse effects on the reproductive system of this organism. These analyses could be useful in developing rapid and efficient chromatin-based genotoxicity assays for pollution biomonitoring programs.