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In this study, a new method has been optimized to determinate nine bisphenols (BPs) in mussel samples. This approach is based on miniaturized matrix solid-phase dispersion combined with gas chromatography-mass spectrometry determination after trimethylsilylation with 99% N,O-Bis-(trimethylsilyl)trifluoroacetamide + 1 % trimethylchlorosilane. The optimization process investigated the derivatization (derivatizing agent volume, time and temperature) and sample preparation (C18 co-column packed and C18 dispersant agent amounts and elution volume) parameters. A second clean-up step was performed using EMR-Lipid cartridges in order to efficiently remove lipids from mussel, the optimized parameters were Captiva EMR-Lipid sorbent amount and elution volume. The whole method was validated and very good linearity (r2 > 0.99) was obtained. The recoveries (accuracy) at two concentration levels ranged from 51.2 to 109 % and the coefficients of variation (precision) were between 0.55 and 13 %. The limits of quantification were between 0.014 μg/Kg dry weight for BPM and 4.0 μg/Kg dry weight for BPS. The analytical procedure was applied to four mussel samples from Galician Rías. Except for one of four samples whose BPAF level was below the limit of quantification, all BPs were found in concentrations that ranged from 0.39 to 301 μg/Kg dry weight.

Metatranscriptomic methods involving RNA sequencing (RNA-seq) are powerful tools for evaluating the gene expression profiles of transcriptionally-active microbes that inhabit the tissues of animals. Bivalve molluscs, like all invertebrates, are holobionts and sites of interactions between host animals and both prokaryotic and eukaryotic symbionts. The present communication describes the metatranscriptomic profile of the resident microorganisms that inhabit the gut of blue mussels, Mytilus edulis, under standard laboratory conditions. Each of the eight mussels described herein were housed in isolated microcosms with routine husbandry for 14 days before their gut tissues were sampled and subjected to RNA-seq. Subsequent mRNA reads that aligned to the mussel genome were removed, and the non-host reads were annotated for function and pathway analyses. Under laboratory conditions, the resident gut microbiota expressed genes associated mostly with aerobic energy processes, with other notable contributions from metabolism and protein processing genes. At the pathway level, the most abundant complete pathways expressed in the resident gut microbial communities were related to aerobic cellular respiration, nucleotide biosynthesis, and catabolism. These data represent novel, baseline microbial gene expression information from the gut of mussels, which are crucial for future research examining the mussel holobiont and bivalve microbial ecology. Public retrieval and secondary analyses of these metatranscriptomic profiles are highly encouraged.

The accumulation of contaminants in aquatic organisms is of concern to human health due to the potential for exposure through the consumption of seafood. This dataset presents the levels of 17 polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and 12 dioxin-like polychlorinated biphenyls (DL-PCBs) (PCB-81, PCB-77, PCB-126, PCB-169, PCB-123, PCB-118, PCB-114, PCB-105, PCB-167, PCB-156, PCB-157 and PCB-189) in bivalves collected annually between 2008 and 2023 from the four Shellfish Production Areas of the Basque coast (southeastern Bay of Biscay). Additionally, data on 6 non-dioxin like polychlorinated biphenyls (NDL-PCBs) (PCB-28, PCB-52, PCB-101, PCB-153, PCB-138 and PCB-180) is provided for the 2012-2023 time period. Depending on the availability of bivalves at each Shellfish Production Area, different species, such as mussels (Mytilus galloprovincialis) and oysters (Magallana gigas and Ostrea edulis) were used for the evaluation. Bivalve samples were analyzed at the accredited Laboratory of Dioxins of the Institute of Environmental Assessment and Water Research - CSIC in Spain, following standardized methodologies according to the specific requirements established at the corresponding EU Regulation. Concentrations and the World Health Organization-Toxic Equivalents (WHO-TEQ) were calculated. Considering the data from the four Shellfish Production Areas and the study period, WHO-TEQ values ranged from 0.12 to 0.64 pg g−1 wet weight for PCDD/Fs and from 0.45 to 2.23 pg g−1 wet weight for DL-PCBs, whereas concentrations ranged from 7.45 to 51.10 ng g−1 wet weight for NDL-PCBs. This database is useful for (i) assessing the spatial and temporal trends of PCDD/Fs and various PCBs in bivalves from the Basque coast, (ii) determining the relative contribution of different congeners, (iii) comparison with levels for human consumption and environmental quality standards, as well as with levels of other biogeographical areas, and (iv) the management of Shellfish Production Areas on the Basque coast.

Mussels can form tough and long-lasting adhesions to organic and inorganic surfaces in saline and impactive severe aquatic environments. Similar to mussel adhesion, dentin bonding occurs in a wet environment. However, unlike mussels, it is difficult to achieve long-lasting bonds with dentin. Moreover, water is considered a major hindrance in dentin bonding. Inspired by the synergistic effect of cationic lysine (Lys) and catechol on the elimination of the hydration layer during mussel adhesion, a catechol- and Lys-functionalized polymerizable polymer (catechol–Lys–methacrylate [CLM]) was synthesized to replicate the complex synergy between amino acids and catechol. The bond-promoting potential of 5 ​mg/mL CLM primer was confirmed using an in vitro wet dentin-bonding model, which was characterized by an improvement in bond strength and durability. CLM can adhere to wet demineralized dentin, with Lys acting as a molecular vanguard to expel water. Subsequently, a myriad of interfacial interactions can be obtained by introducing the catechol group into the interface. Additionally, tough and long-lasting adhesion, similar to that formed by mussels, can be achieved by grafting CLM onto type I collagen via covalent bonds, hydrogen bonds, Van der Waals interactions, and cation–π interactions, which can enhance the mechanical and chemical stability of collagen, increase the enzymatic resistance of collagen, and provide additional physical/chemical adhesion to dentin bonds. Catechol- and cationic Lys-functionalized polymers can improve the stability of the resin–dentin interface under wet conditions.

Polystyrene nanoplastics (PS-NPs) are typical accumulated nanoplastics in the marine environment and organisms, and have strong potential risks to marine ecological environment and human health. MiRNAs could respond to and participate in the response process of environmental stressors. However, the response of miRNAs to nanoplastics has not been fully explored. In this study, miRNA responses of digestive glands in mussels Mytilus galloprovincialis treated by 200 nm PS-NPs (20, 200, 2000 μg/L) for 7 days were characterized by BGISEQ-500 deep sequencing and bioinformatics analysis, along with histopathological quantification with planimetric parameters on hematoxylin and eosin (H&E) staining. Results showed that one novel miRNA (novel_mir63) and seven known miRNAs (miR-34_2, miR-34_5, miR-281_8, let-7–5p_6, miR-10, miR-124, miR-29b-3p) were significantly (adjusted P-value < 0.05) differentially expressed after PS-NPs treatments, and most of them were down-regulated expect for novel_mir63 and miR-34_2. Function analysis of target genes corresponding to these differentially expressed miRNAs indicated that PS-NPs disturbed the process related to metabolism, aging, cardiac function, neural excitation, and repairment. Among them, acetyl-CoA C-acetyltransferase and purine metabolism pathway played vital connection roles. Meanwhile, significantly morphology changes of digestive tubes obtained from H&E stained sections also implied severely disrupted metabolic capability in digestive glands, reflected by significantly increased mean diverticular radius (MDR) and mean luminal radius (MLR) values and the ratio of MLR to mean epithelial thickness (MET), and significantly decreased MET value and MET/MDR. Overall, these findings have revealed new characterization of miRNAs and their target genes in mussel M. galloprovincialis under PS-NPs stress, and provide important clues to further elucidate the toxicity mechanisms of PS-NPs.

In the marine environment, various weathering effects on micro or nanoplastics lead to surface modifications, which in turn alter their toxic effects on aquatic organisms. This study investigated the impact of three types of nanoplastics (NPs, NPs-NH 2 , NPs-COOH) on the antioxidant capacity of Mytilus coruscus gills, mantle, and hemolymph over 28 days. Analyzed key antioxidant stress indicators (CAT, SOD, GSH, GSH-Px, MDA, H 2 O 2 ) and conducted IBR and PCA analyses to evaluate the toxic effects of modified nanoplastics. In particular, NPs-NH 2 showed the most significant inhibition of antioxidant enzymes like CAT and GSH-Px in gills and mantle, while NPs-COOH affected a wider range of oxidative stress markers. Furthermore, tissue-specific responses were observed, with gills being the most sensitive to biomarker changes. Overall, NPs-NH 2 emerged as the most toxic nanoplastic, highlighting the need to assess ecological risks associated with novel nanoparticles in marine environments and offering insights into tissue-specific toxicity in mussels., 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 Elsevier Ltd. All rights reserved.)

The extensive presence of microplastics (MPs) in marine ecosystems constitutes a major threat to aquatic environments. The gametes of the marine invertebrate Mytilus galloprovincialis, which is essential for coastal ecosystems, are released directly into the water, potentially exposing them to environmental microplastics (EMPs). This study examined the effects of exposing M. galloprovincialis gametes to 50 or 100 µg/L EMP for 1 h on fertilization rates, larval quality, and the molecular mechanisms underlying the induction of apoptosis and shell growth. Our findings show that increased EMP concentrations correlate with reduced fertilization success and higher rates of larval malformations, indicating negative impacts on embryonic development. Additionally, DNA degradation in larvae is related to the EMP concentration. The apoptosis-associated proteins Bax, P53, and Cas-3 are upregulated, whereas Bcl-2 and DNA-ligase are downregulated with increasing EMP concentrations. Prothymosin-ɑ (PTMA), which is crucial for cell proliferation, also decreases with increasing EMP concentrations, contributing to impaired cell proliferation and growth imbalances. Reduced HRG gene expression is correlated with decreased shell growth and larval malformations. This study underscores the detrimental impact of EMPs on bivalve gametes, which impacts fertilization success and larval quality and highlights the potential risks to species survival and marine ecosystem stability., 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 Authors. Published by Elsevier B.V. All rights reserved.)

Growing micro- and nano-plastic (MNPs) pollution in the environment poses a threat to marine animals. Due to their excellent filtration capacity, bivalves can easily ingest MNPs, which could be translocated to open circulation system with potential risks. In the present study, the accumulation and elimination of MNPs (200 nm and 1 µm) in the mussel hemolymph serum and hemocytes were firstly quantified, and the differential sensitiveresponses of two subpopulations of hemocytes were then explored by in vivo exposure under environmentally relevant concentration of MNPs (200 µg/L). We demonstrated that MNPs were readily translocated into hemolymph serum, but were immediately followed by efficient internalization by hemocytes. Remarkably, concentrations of MNPs in hemolymph were only 0.63 and 0.39 times lower than the ambient exposure concentration. Granulocytes displayed a much higher potential of accumulating MNPs than the agranulocytes. MPs were more readily internalized by granulocytes, with their estimated maximum bioaccumulation factor (BCF) of 0.29 L/g. Due to the primary function of phagocytic encapsulation of MNPs by granulocytes, lysosome features especially the decline of subsequent lysosome membrane potential could be a potential sensitive biomarker in response to MNPs exposure. Our results provided insights on the bioaccumulation of MNPs at the cellular levels in marine bivalves., 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 Elsevier B.V. All rights reserved.)

This study investigates the applicability of elemental and Cu isotope compositions in sediments and bivalves from the Korean coast to monitor anthropogenic Cu contamination. Sediments with high Cu (>64.4 mg/kg) and/or moderate enrichment levels (EF Cu ) exhibit homogenous δ 65 Cu AE647 values (-0.12 to +0.16 ‰), suggesting similar anthropogenic Cu fingerprints along the Korean coast. Sediments with Cu concentrations near natural background levels (< 20.6 mg/kg) display large isotopic variability (Δ 65 Cu max-mim : ~0.8 ‰), encompassing those from sediments under anthropic influences. We hypothesize that Cu isotopic compositions of Korean geology are heterogeneous, therefore, natural end-members of source mixing models should be established locally at small scales. Cu concentrations in Oysters correlate with sediments, and their isotopic compositions are more suitable for monitoring Cu contamination, while mussel's regulatory mechanisms seem to affect source records. The current Cu isotope data will help to detect shifts attributable to anthropic contamination in future biomonitoring., 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 Elsevier Ltd. All rights reserved.)

Biomineralisation of bivalve shells raises questions at the level of genes to the final calcified product. For the first time, gene expression has been studied in association with growth increment deposition in the mussel Mytilus galloprovincialis. A short-term experiment highlighted that biomineralisation genes exhibit a rhythm of expression consistent with the observed tidal increment formation. Long-term mark-recapture experiments were conducted in three Mediterranean environments and revealed that the mussel shells harbour complex incrementation regimes, consisting of daily, tidal and a mixed periodicity of ∼1.7 growth increment.d -1 formed. The latter is likely related to the local tidal regime, although the mussels were continuously submerged and exposed to a small tidal range. The pattern of growth increments shifted from mixed to daily in Mediterranean lagoon, and to tidal at sea, probably linked to biological clocks. Based on our results and the literature, a hypothetical model for mussel shell increment formation in various habitats is proposed., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Franck Lartaud and Laurence Besseau reports financial support was provided by Centre National de la Recherche Scientifique. If there are other authors, they 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 Authors. Published by Elsevier Ltd.. All rights reserved.)

Microplastics (MPs) have accumulated in the oceans, causing adverse effects on marine organisms and the environment. Biodegradable polylactic acid (PLA) is considered as an excellent substitute for traditional petroleum-based plastics, but it is difficult to degrade completely and easily become MPs in the marine environment. To test the ecological risk of bio-based PLA, we exposed thick-shelled mussels (Mytilus coruscus) to bio-based PLA and petroleum-based polystyrene (PS) (at 10 2 , 10 4 , and 10 6 particles/L) for 14 days. The significant increase in enzyme activities related to oxidative stress and immune response showed that mussels were under physiological stress after MP ingestion. While enzyme activities of nerve conduction and energy metabolism were significantly disturbed after exposure. Meanwhile, normal physiological activities in respiration, ingestion and assimilation were also suppressed in association with enzyme changes. The negative effects of PS and PLA in mussels were not differentiated, and further integration analysis of integrated biomarker response (IBR) and principal component analysis (PCA) also showed that PLA would induce adverse effects in mussels and ecological risks as PS, especially at environmental concentrations. Therefore, it is necessary to pay more attention to the environmental and ecological risk of bio-based MP PLA accumulating in the marine environment., 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 Elsevier B.V. All rights reserved.)

The objective of this work was to determine the spatial distribution and temporal trend (2010–2019) of organochlorine compounds (OCs), polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs), in wild mussels from several Galician Rías (estuaries). Some of them, like the northernmost estuaries, were studied for the first time. OCs were determined by GC-ECD and confirmed by GC-MS/MS. SPCB levels (sum of PCBs 31, 28, 52, 101, 118, 153, 105, 138, 156, 180) were found in the range 7.2–263.1 ng/g dw (dry weight) and SDDt levels (sum of pp´- DDT, op´-DDT, pp´-DDE and pp´-DDD) in the range 2.5–18.8 ng/g dw. PCB levels were below European limits. Analysis of variance (ANOVA) showed some relationship between of OC levels and biological parameters (lipid content and condition index). Principal component analysis (PCA) reported a geographical characterization of Rías Altas and Rías Baixas and relationships between PCB congeners that revealed a similar pattern of entry. Legal restrictions on the production and use of PCBs and OCPs have led to a steady decline in OCs over the period under study (2010–2019).

Coastal waters face significant anthropogenic stress, particularly from tourism, exacerbating pollution, especially in areas like touristic islands. Ischia, the largest island in the Gulf of Naples and part of the Regno di Nettuno Marine Protected Area, suffers from pollution due to tourism and maritime traffic. During the initial SARS-CoV-2 lockdown from March to June 2020, Ischia was isolated, providing a unique opportunity to study pollutant release and its impact on coastal ecosystems. Adult Mytilus galloprovincialis mussels were transplanted to three sites on the island for active biomonitoring. Accumulation of chemicals in tissues and biomarkers related to metabolism, detoxification, and oxidative stress were measured. Results indicated that pollutants from daily activities entered the sea, affecting filter feeders. Translocated organisms showed modulated metabolic functions and biochemical changes, highlighting coastal vulnerability and calling for conservation efforts., 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 Authors. Published by Elsevier Ltd.. All rights reserved.)

Numerous approaches have been developed to improve the resin-dentin bond performance, among which the bio-application of mussel-derived compounds have drawn great attention recently. To assess the performance of N-(3,4-dihydroxyphenethyl)methacrylamide (DMA), a mussel-derived compound, as a functional monomer in dental adhesive, its potential property to cross-link with dentin collagen and polymerize with adhesive will first be evaluated by transmission electron microscopy (TEM), attenuated total reflectance technique of Fourier transform infrared (ATR-FTIR), and atomic force microscopy (AFM) via Peakforce QNM mode. After validating the influence of DMA on collagen and adhesive separately, the overall performance of DMA/ethanol solution as a primer in dentin bonding was examined using micro-tensile bond strength (μTBS) testing, fracture pattern observation, and nanoleakage evaluation both immediately and after 10,000 times thermocycling aging. The inhibitory effect of DMA on endogenous metalloproteinases (MMPs) was evaluated by in situ zymography using confocal laser scanning microscopy (CLSM) and the cytotoxicity of DMA was evaluated using cell counting kit-8. Results demonstrated that DMA successfully cross-linked with dentin collagen via non-covalent bonds and had no influence on the polymerization and mechanical properties of the adhesive. Furthermore, even after 10,000 times thermocycling aging, the μTBS and nanoleakage expression of the DMA-treated groups showed no significant change compared with their immediate values. In situ zymography revealed reduced endogenous proteolytic activities after the application of DMA, and no cytotoxicity effect was observed for DMA concentration up to 25 ​μmol/L. Thus, DMA could be used as a novel, biocompatible functional monomer in dentin bonding.

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.

Perfluorooctanoate (PFOA) is widely used as a surfactant and has metabolic, immunologic, developmental, and genetic toxicity on marine organisms. However, the effects of PFOA on individual defense functions in mussels in the presence of titanium dioxide nanoparticles (nano-TiO 2 ) are poorly understood. To investigate the defense strategies and regulatory mechanisms of mussels under combined stressors, the thick-shell mussels Mytilus coruscus were exposed to different PFOA concentrations (0, 2 and 200 μg/L) and nano-TiO 2 (0 and 0.1 mg /L, size: 25 nm) for 14 days. The results showed that, compared to the control group, PFOA and nano-TiO 2 significantly reduced the number of byssal threads (NBT), byssal threads length (BTL), diameter of proximal threads (DPB), diameter of middle threads (DMB), diameter of distal byssal threads (DDB), adhesive plaque area (BPA), and breaking force of byssal threads (N). Under the influence of PFOA and nano-TiO 2 , the morphological surface smoothness of the fractured byssal threads surface increased, concurrently inducing an increased surface roughness in the adhesive plaques. Additionally, under the presence of PFOA and nano-TiO 2 , the foot displayed dispersed tissue organization and damaged villi, accompanied by an increased incidence of cellular apoptosis and an upregulation of the apoptosis gene caspase-8. Expression of the adhesion gene mfp-3 and byssal threads strength genes (preCOL-D, preCOL-NG) was upregulated. An interactive effect on the performance of byssal threads is observed under the combined influence of PFOA and nano-TiO 2 . Under co-exposure to PFOA and nano-TiO 2 , the performance of the byssal threads deteriorates, the foot structure is impaired, and the genes mRNA expression of byssal thread secretory proteins have compensated for the adhesion and byssal threads strength by up-regulation. Within marine ecosystems, organic and particulate contaminants exert a pronounced effect on the essential life processes of individual organisms, thereby jeopardizing their ecological niche within community assemblages and perturbing the dynamic equilibrium of the overarching ecosystem. ENVIRONMENTAL IMPLICATION: Perfluorooctanoic acid (PFOA) is prone to accumulate in marine organisms. TiO 2 nanoparticles (nano-TiO 2 ) are emerging environmental pollutants frequently found in marine environment. The effects of PFOA and nano-TiO 2 on marine mussels are not well understood, and their toxic mechanisms remain largely unknown. We investigated the impacts of PFOA and nano-TiO 2 on mussel byssus defense mechanisms. By assessing byssus performance indicators, morphological structures of the byssus, subcellular localization, and changes in byssal secretion-related genes, we revealed the combined effects and mechanisms through which these two types of pollutants may affect the functional capabilities and survival of mussels in the complex marine ecosystem., 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 Elsevier B.V. All rights reserved.)

Tetrabromobisphenol A (TBBPA) and Perfluorooctane sulfonate (PFOS) are emerging contaminants which coexist in marine environments, posing significant risks to ecosystems and human health. The behavior of these contaminants in the presence of dissolved organic matter (DOM), specifically the co-contamination of TBBPA and PFOS, is not well understood. The bioaccumulation, distribution, elimination, and toxic effects of TBBPA and PFOS on thick-shell mussels (Mytilus unguiculatus V.), with the absence and presence of humic acid (HA), a typical DOM, were studied. The results showed that the uptake of TBBPA decreased and the uptake of PFOS increased when exposed to 1 mg/L HA. However, at higher concentrations of HA (5 and 25 mg/L), the opposite effect was observed. Combined exposure to HA, TBBPA, and PFOS resulted in oxidative stress in the digestive gland, with the severity of stress dependent on exposure time and HA dose. Histological analysis revealed a positive correlation between HA concentration and tissue damage caused by TBBPA and PFOS. This study provides insights into the influence of HA on the bioaccumulation-elimination patterns and toxicity of TBBPA and PFOS in marine bivalves, offering valuable data for ecological and health risk assessments of combined pollutants in aquatic environments rich in DOM., 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 Elsevier B.V. All rights reserved.)

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., 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 Authors. Published by Elsevier Ltd.. All rights reserved.)

This study aimed to identify the presence of microplastics in green mussels (Perna viridis), surface seawater, and beach sediment on the North Coast of Vietnam. The average concentration of MPs in mussels was 3.67 ± 1.20 MPs/g wet weight and 25.05 ± 5.36 MPs/individual. Regarding surface seawater and beach sediments, the MPs concentration was found at 88.00 ± 30.88 MPs/L and 4800 ± 1776 MPs/kg dry weight, respectively. The dominant microplastics shape was fragment with the fractions ranging from 69.86 to 82.41 %. In addition, the size distribution of MPs was mostly in the range of smaller than 50 μm and 1-150 μm (34.17 % and 45.62 % in mussels; 29.65 % and 43.20 % in surface seawater and 40.22 % and 39.40 % in beach sediment, respectively). Polyethylene terephthalate was the major polymer types 49.93-58.44 % of the detected MPs. The risk assessment results based on the polymer types indicated a warning level in several sites., 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 Elsevier Ltd. All rights reserved.)

Santa Catarina is the main producer state of oysters and mussels in Brazil, reaching 98 % of national production. To assure the safety of bivalve mollusks production, control programs of marine biotoxins (MBs) have been continuously performed. Herein, the co-occurrence of MBs and contaminants of emerging concern (CECs) in oyster and mussels from the main production sites of Santa Catarina was reported, covering 178 compounds. Samples of wild and non-cultivated oysters and mussels were also assessed. Chemometric tools were used to evaluate and optimize several sample preparation techniques such as solid-liquid, ultrasound assisted, and pressurized liquid extraction. The optimized protocol was based on ultrasound assisted extraction followed by liquid chromatography coupled to tandem mass spectrometry. The results showed the incidence of several CECs and MBs. In the case of MBs, all results were below the regulatory limits for both cultivated and non-cultivated samples. Wild mollusks have shown a higher number of compounds. Regarding CECs, the more frequent compounds were caffeine, diclofenac, meloxicam, and sertraline. Domoic acid and okadaic acid were the main toxins detected. The results highlighted the need of monitoring for MBs and the potential of oyster and mussels as sentinel organisms to risk analysis of CECs in coastal regions. To the best of our knowledge, this is the first method to describe a simultaneous sample preparation and analysis of CECs and MBs in bivalve mollusks, as well as the first report of meloxicam and florfenicol in mussels and oysters., 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 © 2023 Elsevier B.V. All rights reserved.)

Nowadays, it is of utmost importance to consider climate change factors, such as ocean warming, since the risk of negative impacts derived from increased surface water temperature is predicted to be high to the biodiversity. The need for renewable energy technologies, to reduce greenhouse gas emissions, has led to the increasing use of rare earth elements (REEs). Dysprosium (Dy) is widely used in magnets, motors, electrical vehicles, and nuclear reactors, being considered a critical REE to technology due to its economic importance and high supply risk. However, the increasing use of this element contributes to the enrichment of anthropogenic REEs in aquatic systems. Nevertheless, the information on the potential toxicity of Dy is limited. Moreover, the effects of pollutants can be amplified when combined with climate change factors. Thus, this study aimed to assess the effects of Dy (10 μg/L) in the species Mytilus galloprovincialis under actual (17 °C) and predicted warming conditions (21 °C). The Dy concentration in contaminated mussels was similar between temperatures, probably due to the detoxification capacity in individuals under these treatments. The combined stressors affected the redox balance, but higher impacts were caused by Dy and warming acting alone. In terms of cellular damage, although Dy acting alone was prejudicial to mussels, warming and both stressors acting together induced higher levels of LPO and PC. The histopathological effects of Dy in the digestive tubules were independent of the temperature tested. Regarding effects on sperm, only warming induced cellular damage, while both stressors, alone and together, impaired sperm movement. Overall, this study highlights that warming might influence the effects induced by Dy, but greater impacts were caused by the element. Eventually, the tested stressors may have consequences on mussels' reproduction capacity as well as their growth, abundance, and survival., 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 © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

Blue mussels (Mytilus sp.) are an economically important species for European aquaculture. Their importance as a food source is expected to increase in the coming net-zero society due to their low environmental footprint; however, their production is affected by anthropogenic stressors and climate change. During reproduction, lipids are key molecules for mussels as they are the main source of energy on which newly hatched embryos depend in the first days of their development. In this work, blue mussels of different origins are analysed, focusing on the differences in lipid composition between the ovary (BMO) and the testis (BMT). The lipidome of blue mussel gonads (BMG) is studied here by combining traditional lipid profiling methods, such as fatty acid and lipid class analysis, with untargeted liquid chromatography-mass spectrometry (LC-MS) lipidomics. The approach used here enabled the identification of 770 lipid molecules from 23 different lipid classes in BMG. BMT, which consists of billions of spermatocytes, had greater amounts of cell membrane and membrane lipid components such as FA18:0, C20 polyunsaturated fatty acids (PUFA), free sterols (ST), ceramide phosphoethanolamines (CerPE), ceramide aminoethylphosphonates (CAEP), cardiolipins (CL), glycerophosphocholines (PC), glycerophosphoethanolamines (PE) and glycerophosphoserines (PS). In BMO, saturated fatty acids (FA14:0 and FA16:0), monounsaturated fatty acids (MUFA) and other storage components such as C18-PUFA accumulated in triradylglycerolipids (TG) and alkyldiacylglycerols (neutral plasmalogens, TG O-), which, together with terpenes, wax esters and cholesterol esters, make up most of oocytes yolk reserves. BMO also had higher levels of ceramides (Cer) and generally alkyl/alkenyl glycerophospholipids (mainly plasmanyl/plasmenyl PC), suggesting a role for these lipids in vitellogenesis. Non-methylene interrupted dienoic fatty acids (NMID FA), typically found in plasmalogens, were the only membrane-forming PUFA predominantly detected in BMO. The results of this study are of great importance for clarifying the lipid composition of BMG and provide an important basis for future studies on the reproductive physiology of these organisms., 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 © 2023 Elsevier Inc. All rights reserved.)

This study investigated the presence of microplastics (MPs) in mussels from an urbanized bay in Korea, focusing on intra-annual variation. The average MP concentration was 0.36 ± 0.14 n/g w.w. (1.24 ± 0.56 n/individual). MPs were detected throughout the year with minor monthly variation and no significant monthly changes. Although there was no correlation between MP concentration in mussel and seawater, a positive relationship was observed when normalizing MPs to mussel's condition index (except for July). Polypropylene, polyethylene and acrylate polymer were dominant MPs, with high-density polymers comprising 59 % of the total. Most MPs were fragments and particles <200 μm. This study indicates that strict time restrictions for mussel sampling may not be necessary to assess MP levels in this bay. Additionally, considering MPs in seawater, biological conditions of mussel may be essential for understanding variations in MP levels in mussels. Including small MPs (<200 μm) and high-density polymers in MP assessment is recommended., 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 © 2023 Elsevier Ltd. All rights reserved.)

The spatial distribution of total mercury (HgT) and methylmercury (MeHg) in Mediterranean mussels (Mytilus galloprovincialis) sampled at 23 different stations along Türkiye's Black Sea, the Marmara, and Aegean Sea coasts were investigated in this study. HgT and MeHg were determined using a direct mercury analyzer and ranged 1.10-130.70 μg/kg and 0.22-37.90 μg/kg dry weight, respectively. HgT and MeHg have been determined to have a substantial positive correlation (r 2  = 0.94). According to average consumption and portion sizes, the weekly intake (EWI) for MeHg was less than the 1.6 μg/kg/week limits set by the FAO/WHO Joint Committee of Experts on Food Additives (JECFA) for both pediatric and adult consumers. The Target Risk Coefficients (THQ) were calculated to explain the potential long-term exposure scenarios. Our data showed that the excessive consumption of mussels from areas with high MeHg concentrations might pose a risk only for pediatric consumers. The determination of suitable sites for healthy-sustainable mussel production as well as the prevention of collecting mussels from illegal and polluted fields and placing them on the market, should be considered in the context of consumer health awareness., 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 © 2023 Elsevier Ltd. All rights reserved.)

Polybrominated diphenyl ethers (PBDEs) are one of the most used halogenated flame retardants worldwide, and exert neurotoxicity, reproductive toxicity, endocrine interference, and carcinogenic effects on organisms. However, there are insufficient studies on the physical and immune defense at the individual level of mussels under different food conditions. To explore the defense strategy and individual health status, the thick-shelled mussels Mytilus coruscus were exposed to different BDE-47 concentrations (0, 0.1 and 10 μg/L) and nutritional conditions (feeding and starvation) for 21 days. The results showed that BDE-47 exposure and starvation significantly decreased the number of byssus threads (NBT), adhesion, and condition index (CI) of mussels, whereas increased the reactive oxygen species (ROS) production and the combined stress further declined the CI. BDE-47 exposure and starvation induced decreased adhesive capability and healthy state along with oxidative lesions in mussels. The downregulation gene expression of foot adhesion proteins (mfp-2/3/4/5/6) under starvation or combined exposure also proved the reduced adhesion of mussels. However, up-regulated mfp-1 and pre-collagens proteins (preCOL-D/P/NG) indicated mussels would adjust energy allocation to enhance the strength and extensibility of byssal threads for compensating reduced adhesion and CI. As global climate change and organic pollution have dramatically impacted the ocean, hazardous substances and the fluctuated primary productivity have frequently co-occurred, which will affect the structure of coastal biomes and fishery production., 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 © 2023 Elsevier B.V. All rights reserved.)

Nano‑titanium dioxide (nTiO 2 ) is a widely used nanomaterial posing potential ecological risk for marine ecosystems that might be enhanced by elevated temperatures such as expected during climate change. nTiO 2 may affect benthic filter feeders like mussels through waterborne exposures and via food chain due to the adsorption on/in algae. Mussel byssus are proteinaceous fibers secreted by byssal glands of the mussels for attachment. Byssus production and mechanical properties are sensitive to environmental stressors but the combined effects of warming and nTiO 2 on byssus performance of mussels are unclear hampering our understanding of the predation and dislodgement risk of mussels under the multiple stressor scenarios. We explored the effects of a short-term (14-day) single and combined exposures to warming (28 °C) and 100 μg L -1 nTiO 2 (including food co-exposure) on the byssus performance of the thick shell mussel Mytilus coruscus. The mechanical strength (measured as the breaking force) of the byssal threads was impaired by warming and nTiO 2 (including food co-exposure), but the number and length of the byssal threads were increased. The mRNA expression levels of mussel foot proteins (mfp-3, mfp-5) and pre-collagens (preCOL-D, preCOL-P, preCOL-NG) were up-regulated to varying degrees, with the strongest effects induced by warming. This indicates that the physiological and molecular mechanisms of byssus secretion are plastic. However, downregulation of the mRNA expression of preCOL-D and preCOL-P under the combined warming and nTiO 2 exposures indicate the limits of these plasticity mechanisms and suggest that the attachment ability and survival of the mussels may be impaired if the pollution or temperature conditions further deteriorate., 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 © 2023 Elsevier B.V. All rights reserved.)

Oil spills pose significant environmental risks, particularly in cold seas. In the Baltic Sea, the low salinity (from 0 to 2 up to 18) affects the behaviour of the spilled oil as well as the efficiency and ecological impacts of oil spill response methods such as mechanical collection and the use of dispersants. In the present study, mussels (Mytilus trossulus) were exposed under winter conditions (5 °C) to the water-accommodated fraction (WAF) of Naphthenic North Atlantic crude oil prepared by mechanical dispersion or to the chemically enhanced fraction (CEWAF) obtained using the dispersant Finasol OSR 51 at salinities of 5.6 and 15.0. Especially at the lower salinity, high bioaccumulation of polycyclic aromatic hydrocarbons was recorded in mussels in the CEWAF treatments, accompanied by increased biomarker responses. In the WAF treatments these impacts were less evident. Thus, the use of dispersants in the Baltic Sea still needs to be carefully considered., 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 © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is currently the gold-standard technique for detecting and quantifying messenger RNA. However, without proper validation, the method may produce artefactual and non-reproducible cycle threshold values generating poor-quality data. The newer droplet digital PCR (ddPCR) method allows for the absolute quantification of targeted nucleic acids providing more sensitive and accurate measurements without requiring external standards. This study compared these two PCR-based methods to measure the expression of well-documented genes used in ecotoxicology studies. We exposed Mediterranean mussels (Mytilus galloprovincialis) to copper and analyzed gene expression in gills and digestive glands using RT-qPCR and ddPCR assays. A step-by-step methodology to optimize and compare the two technologies is described. After ten-fold serial complementary DNA dilution, both RT-qPCR and ddPCR exhibited comparable linearity and efficiency and produced statistically similar results. We conclude that ddPCR is a suitable method to assess gene expression in an ecotoxicological context. However, RT-qPCR has a shorter processing time and remains more cost-effective., 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 © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Intertidal mussels are well adapted to withstand emersion from water during low tide, but they may be intermittently exposed to waterborne toxicants such as copper, which targets physiological processes including metabolism, ammonia excretion, and osmoregulation. To determine if copper exposure damages intertidal organisms' ability to tolerate tidal emersion, Mediterranean mussels (Mytilus galloprovincialis) were exposed to copper for 96 h followed by 6 h of emersion. Oxygen uptake increased after copper exposure which suggests that copper accumulation caused moderate stress in the mussels, but ammonia excretion and anaerobic metabolism were unaffected by mixed copper and emersion exposures. Shell composition analyses indicate that cycles of copper exposure and tidal emersion may affect bivalve shell growth, but copper deposition into shells may decrease the metal's overall toxicity. Results suggest that copper does not damage M. galloprovincialis's tolerance to tidal emersion, and insight is provided into the mussel's ability to overcome mixed stressor exposures., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Tamzin Blewett reports financial support was provided by Natural Sciences and Engineering Research Council of Canada., (Copyright © 2023 Elsevier Ltd. All rights reserved.)