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Farmed mussels accumulate contaminants from their production environment rather than releasing them into water. This study reveals potential health risks associated with selenium, cadmium, mercury, and lead resulting from the consumption of mussels (Mytilus galloprovincialis) cultured along the coasts of Türkiye and Bulgaria. The concentrations of Se and toxic trace metals were measured by inductively coupled plasma mass spectrometry (ICP-MS). The detection limits (LOD) were 0.100, 0.015, 0.025, and 0.180 µg/kg for Se, Cd, Hg, and Pb, respectively. The mean Se concentrations were between 1.305 and 1.957 µg/g, and toxic metals were below the maximum limits. Due to Turkish and Bulgarian consumers' limited mollusk consumption, mussels could only provide a maximum of 7.35% of the daily Se need. THQ and TTHQ of Se, Cd, and methyl-Hg were below 1, indicating that farmed mussels were safe for consumption. Percent PTWI values were calculated only for Cd and MeHg, as the PTWI value for Pb was discarded by the authorities and not determined for Se. Accordingly, weekly mussel consumption did not pose any risks. The margin of exposure approach was used to evaluate Pb intake. MOE-SBP and MOE-NE were significantly higher than 10, designating no significant health risks. Long-term consumption of mussels also does not pose a carcinogenic risk regarding the TR index calculated between 10 -5 and 10 -6 for Pb. Positive HBV Se (10.13-37.27) indicated that Se in mussels overcame Hg-related potential health concerns. Consequently, mussels grown in Türkiye and Bulgaria did not pose a risk for human consumption, based on current risk analysis methods., (© 2024. The Author(s).)

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

The aim of this study was to evaluate the potential bioaccumulation of arsenic (As), copper (Cu), zinc (Zn), cadmium (Cd), lead (Pb) and mercury (Hg) in the haemolymph and corpus of Mytilus galloprovincialis and Tapes decussatus from Lake Faro. The lake is particularly prone to the accumulation of substances that are potentially toxic to aquatic organisms, due to the input of pollutants from urban and agricultural sources and the low rate of water exchange. The combination of saltwater from the Tyrrhenian Sea, the Strait of Messina and freshwater from hilly aquifers has created brackish conditions in the lake, resulting in an area of high commercial shellfish productivity. As, Cd, Cu, Pb and Zn were determined using a single quadrupole inductively coupled plasma mass spectrometer; Hg was determined using a direct mercury analyser (DMA-80). Physicochemical parameters of the water from Lake Faro were also performed. Statistical analysis was carried out using GraphPad Prism 9.0 (GraphPad Software, Inc., Boston, MA, USA) and Shapiro-Wilk normality was applied. Concentrations of Cd, Hg and Pb below the permitted MRLs in Mytilus galloprovincialis and Tapes decussatus used as ''biological indicators'' show that Lake Faro is not at risk of contamination by these pollutants and, moreover, is free of health problems for the consumer based on regulatory limits., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Mussel byssal proteins are of biomimetic importance for the development of novel underwater bio-adhesive agents. It is important to maintain a reduced state during the process of byssus adhesion. There are 19 mussel foot proteins (MFPs) have been reported in previous studies, among which only MFP-6 had been confirmed as an antioxidant protein in mussel byssus due to the function of cysteines, and playing an essential role in the redox balance of mussel byssus during adhesion process. Although the other four MFPs (MFP-16 ~ MFP-19) also have abundant cysteines, their function is still unknown. In this study, a novel mussel foot protein, named MFP-20, was identified from Mytilus coruscus foot. The sequential features, expression profile, and function of recombinant MFP-20 were verified. The results showed that MFP-20 has more abundant cysteines than other MFPs, the relative expression of mfp-20 was upregulated in Fe 3+ stress and low pH seawater. In addition, different adhesive substrates induced significant changes of expression level of mfp-20. Furthermore, rMFP-20 showed strong antioxidant capacity in the DPPH assay, and the abundant cysteines in its sequence may play vital roles in the antioxidation activity. Our findings revealed the possible function of MFP-20 with a totally different sequence from the reported MFP-6 and provided new clues for exploring the redox balance of mussel byssus during the adhesion process., 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. Published by Elsevier B.V.)

Uranium is a natural radioelement (also a model for heavier actinides), but may be released through anthropogenic activities. In order to assess its environmental impact in a given ecosystem, such as the marine system, it is essential to understand its distribution and speciation, and also to quantify its bioaccumulation. Our objective was to improve our understanding of the transfer and accumulation of uranium in marine biota with mussels taken here as sentinel species because of their sedentary nature and ability to filter seawater. We report here on the investigation of uranium accumulation, speciation, and localization in Mytilus galloprovincialis using a combination of several analytical (Inductively Coupled Plasma Mass Spectrometry, ICP-MS), spectroscopic (X ray Absorption Spectroscopy, XAS, Time Resolved Laser Induced Fluorescence Spectroscopy, TRLIFS), and imaging (Transmission Electron Microscopy, TEM, μ-XAS, Secondary Ion Mass Spectrometry, SIMS) techniques. Two cohorts of mussels from the Toulon Naval Base and the Villefranche-sur-Mer location were studied. The measurement of uranium Concentration Factor (CF) values show a clear trend in the organs of M. galloprovincialis: hepatopancreas ≫ gill > body ≥ mantle > foot. Although CF values for the entire mussel are comparable for TNB and VFM, hepatopancreas values show a significant increase in those from Toulon versus Villefranche-sur-Mer. Two organs of interest were selected for further spectroscopic investigations: the byssus and the hepatopancreas. In both cases, U(VI) (uranyl) is accumulated in a diffuse pattern, most probably linked to protein complexing functions, with the absence of a condensed phase. While such speciation studies on marine organisms can be challenging, they are an essential step for deciphering the impact of metallic radionuclides on the marine biota in the case of accidental release. Following our assumptions on uranyl speciation in both byssus and hepatopancreas, further steps will include the inventory and identification of the proteins or metabolites involved., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: DEN AUWER Christophe reports financial support was provided by French Alternative Energies and Atomic Energy Commission. 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 Elsevier Inc. All rights reserved.)

Microplastic (MP) pollution has become a global concern due to its potential impacts on the environment, ecosystem services and human health. The goals of the present study were to document the MP contamination in wild specimens of Mytilus galloprovincialis sampled along the Atlantic coast of the North region of Portugal continental (NW Portuguese coast), and to estimate the human risk of MP intake (HRI) through the consumption of local mussels as seafood. Mussels were collected at four sampling sites along the NW Portuguese coast (40 mussels per site), and the whole soft body of each mussel was analysed for MP content. HRI estimates were based on the mean of MP items per wet weight of mussel analysed tissue (MP/g) and consumption habits. A total of 132 MP items were recovered from mussels. MP had diverse sizes (98-2690 μm) and colours. The most common shapes were fibres (39%) and pellets (36%). Five polymers were identified in the MP: polyethylene (50%), polystyrene (15%), poly(ethylene vinyl acetate) (14%), polyamide (12%) and polypropylene (9%). From the 160 analysed mussels, 55% had MP. The mean and standard error of the mean of mussel contamination ranged from 0.206 ± 0.067 and 0.709 ± 0.095 MP/g. Compared to estimates based on MP contamination in mussels from other areas and varied consumption habits, the HRI through the consumption of mussels from the NW Portuguese coast is relatively low., 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.)

The observation of mortality in Mediterranean mussels (Mytilus galloprovincialis) distributed in the Çanakkale Strait in recent years was influential in developing the research question for this study. In this study, the presence of bacteria (Vibrio spp.) and parasites (Marteilia spp. and Haplosporidium spp.) in mussels collected from Kumkale, Kepez, and Umurbey stations in the Çanakkale Strait was investigated seasonally. Microbiological findings, histopathology, oxidative stress enzymes and their gene expressions, lipid peroxidation, lysosomal membrane stability, and changes in haemolymph were examined. In summer samples, both the defence system and the extent of damage were higher in gill tissue. In winter samples, enzyme activities and lipid peroxidation were found to be predominantly higher in digestive gland tissues. Histological examinations and Hemacolor staining revealed the presence of protozoan cysts, and for bacterial examination, molecular analysis performed after culturing revealed the presence of 7 Vibrio species. While the total numbers of heterotrophic bacteria detected in all samples were at acceptable levels, the predominance of Vibrio spp. numbers among the total heterotrophic bacteria detected in almost all samples were noteworthy. The total hemocyte count was calculated as 5.810(4)±0.58 (cells/mm 3 ) in winter and 7.210(4)±1.03 (cells/mm 3 ) in summer. These factors are considered to be possible causes of mussel mortality., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Mert Gurkan reports financial support was provided by SCIENTIFIC AND TECHNOLOGICAL RESEARCH COUNCIL OF Türkiye. 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 Elsevier Ltd. All rights reserved.)

We examined the presence of microplastics in blue mussels Mytilus spp. from the intertidal zone of western Spitsbergen in Arctic Svalbard. The optical microscopy technique detected a total of 148 microplastics, with the highest concentration per mussel being 24 particles. Microplastics were found in 84% of the examined mussels. The microplastics ranged in size from <0.5 mm to 5 mm and consisted of fibers (83%), fragments (13%), plates (3%), and spherules (1%). The micro-Raman spectroscopy technique revealed four different types of polymers: polyethylene (67%), nylon-12 (17%), low-density polyethylene (11%), and polypropylene (5%). Our research shows that Arctic coastal waters are polluted with microplastics notwithstanding their remoteness. These findings suggest that microplastic contamination may harm marine life and coastal ecosystems and require further research into long-term environmental effects. We also indicate that intertidal mussels may be beneficial for monitoring microplastics because they can be collected without involving diving., 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.)

Microplastics are a global ecological concern due to their potential risk to wildlife and human health. Animals ingest microplastics, which can enter the trophic chain and ultimately impact human well-being. The ingestion of microplastics can cause physical and chemical damage to the animals' digestive systems, affecting their health. To estimate the risk to ecosystems and human health, it is crucial to understand the accumulation and localization of ingested microplastics within the cells and tissues of living organisms. However, analyzing this issue is challenging due to the risk of sample contamination, given the ubiquity of microplastics. Here, an analytical approach is employed to confirm the internalization of microplastics in cryogenic cross-sections of mussel tissue. Using 3D Raman confocal microscopy in combination with chemometrics, microplastics measuring 1 μm in size were detected. The results were further validated using optical and fluorescence microscopy. The findings revealed evidence of microplastics being internalized in the digestive epithelial tissues of exposed mussels (Mytilus galloprovincialis), specifically within the digestive cells forming digestive alveoli. This study highlights the need to investigate the internalization of microplastics in organisms like mussels, as it helps us understand the potential risks they pose to aquatic biota and ultimately to human health. By employing advanced imaging techniques, challenges associated with sample contamination can be overcome and valuable insights into the impact of microplastics on marine ecosystems and human consumers are provided., 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.)

Characterized by a sessile filter-feeding lifestyle, commercial marine bivalves inhabiting pollution-prone coastal areas may accumulate significant amounts of pollutants, such as antibiotic residues, in their soft tissues and thus pose a potential threat to the health of seafood consumers. Microplastics are another type of emerging pollutant that are prevalent in coastal areas and have been reported to interact with common antibiotics such as enrofloxacin (ENR) and trimethoprim (TMP). Nevertheless, little is known about the impacts of MPs on the accumulation and corresponding food safety risk of antibiotics in edible bivalve species. Taking the frequently detected ENR, TMP, and polystyrene (PS)-MPs as representatives, the accumulation of above-mentioned antibiotics in three commercial bivalves with or without the copresence of MPs was assessed. In addition, the corresponding food safety risks of consuming antibiotic-contaminated bivalves were evaluated. Moreover, the impacts of these pollutants on detoxification-related processes were analyzed using the thick-shell mussel as a representative. Our results demonstrated that blood clams (Tegillarca granosa), thick-shell mussels (Mytilus coruscus), and Asiatic hard clams (Meretrix meretrix) accumulated significantly higher amounts of antibiotics in their bodies under antibiotic-MP coexposure scenarios. Although based on the target hazard quotients (THQs) and the margins of exposure (MoEs) obtained, the direct toxic risks of consuming ENR- or TMP-contaminated bivalves were negligible, the TMP residue accumulated in TMP-MP-coexposed mussels did surpass the maximum residue limits (MRLs) of the corresponding National Food Safety Standard of China, suggesting that other forms of potential risks should not be ignored. In addition, it was shown that the detoxification, energy provision, and antioxidant capacities of the thick-shell mussels were significantly hampered by exposure to the pollutants. In general, our data indicate that MPs may aggravate the accumulation and corresponding food safety risk of antibiotics in edible bivalves by disrupting detoxification-related processes, which deserves closer attention., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Copyright © 2023 Elsevier B.V. All rights reserved.)

MytiLec-1, the recombinant form of a mussel lectin from Mytillus galloprovincialis, was purified by affinity chromatography and showed the maximum hemagglutination activity at a temperature range of 10 °C to 40 °C and at pH 7.0 to 9.0. Denaturants like urea and acidic-guanidine inhibited its hemagglutination activity significantly. MytiLec-1 was found to be metal-independent though Ca 2+ slightly increased the activity of chelated MytiLec-1. The lectin suppressed 65 % growth of Pseudomonas aeruginosa (ATCC 47085) at 200 μg/ml and reduced the formation of biofilm (15 % at 200 μg/ml). Comparing to Shigella sonnei (ATCC 29930), Shigella boydii (ATCC 231903) and Shigella dysenteriae (ATCC 238135), Bacillus cereus (ATCC 14579) was slightly more sensitive to MytiLec-1. At a concentration of 200 μg/disc and 100 μg/ml, MytiLec-1 prevented the growth of Aspergillus niger and agglutinated the spores of Aspergillus niger and Trichoderma reesei, respectively. Amino acid sequences, physicochemical properties and antimicrobial activities of MytiLec-1 were compared with three other lectins (CGL, MTL and MCL from Crenomytilus grayanus, Mytilus trossulas and Mytilus californianus, respectively) from the mytilectin family of bivalve mollusks. It reconfirms the function of these lectins to recognize pathogens and perform important roles in innate immune response of 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 article., (Copyright © 2023 Elsevier B.V. All rights reserved.)

This study documented seasonal levels of microplastics (MPs) and biomarkers (condition index, neurotoxicity, energy, oxidative stress) in mussels (Mytilus galloprovincialis), and water physico-chemical parameters in the Douro estuary (NE Atlantic coast), and estimated the human risk of MP intake (HRI) through mussels. Mussel stress was determined through the Integrated Biomarker Response (IBR). HRI was estimated from mussel MP concentrations and consumer habits. MPs were mainly micro-fibres (72 %) with varied chemical composition. Seasonal MP means (±SEM) in mussels ranged from 0.111 ± 0.044 (spring) to 0.312 ± 0.092 MPs/g (summer). Seasonal variations of mussel stress (IBR: 1.4 spring to 9.7 summer) and MP concentrations were not related. MeO-BDEs, PBDEs, temperature, salinity and other factors likely contributed to mussel stress variation. HRI ranged from 2438 to 2650 MPs/year. Compared to the literature, MP contamination in mussels is low, as well as the human risk of MP intake through their consumption., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2023. Published by Elsevier Ltd.)

Micronutrients, or essential trace elements, are important components in various metabolic processes inherent to the normal functioning of organism. To date, a substantial part of the world population suffers from a lack of micronutrients in the diet. Mussels are an important and cheap source of nutrients, which can be utilized to mitigate the micronutrient deficiency in the world. In the present work, using inductively coupled plasma mass spectrometry, the contents of the micronutrients Cr, Fe, Cu, Zn, Se, I, and Mo were studied for the first time in soft tissues, shell liquor, and byssus of females and males of the mussel Mytilus galloprovincialis as the promising sources of essential elements in the human diet. Fe, Zn, and I were the most abundant micronutrients in the three body parts. Significant sex-related differences in the body parts were detected only for Fe, which was more abundant in byssus of males, and Zn, which exhibited higher levels in shell liquor of females. Significant tissue-related differences were registered in the contents of all the elements under study. M. galloprovincialis meat was characterized as the optimal source of I and Se for covering the daily human needs. Regardless of sex, byssus turned out to be richer in Fe, I, Cu, Cr, and Mo in comparison with soft tissues, which fact allows recommending this body part for the preparation of dietary supplements to compensate for the deficiency of these micronutrients in the human body., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The release of hazardous chemicals into aquatic environments has long been a known problem, but its full impact has only recently been realized. This study presents a validated liquid chromatography-mass spectrometry (HPLC-MS/MS) method for detecting pharmaceutical and pesticide residues in mussels (Mytilus galloprovincialis). An innovative MS-compatible extraction method was developed and validated, demonstrating successful recovery rates for analytes at three different concentration levels (25-95%). The method detected the target analytes at ng/g concentrations with high accuracy (-7% to 11%) and low relative standard deviation (<10%) for both intra-day and inter-day analyses. After validation, the method was applied to mussel samples collected from a commercial farm near Senigallia, Adriatic Sea, detecting different contaminants in the range of 2-40 ng/g (dry weight). The study provides a valuable tool for investigating the potential threats posed by diverse contaminant classes with high annual tonnage, including analytes with known persistence and/or illegal status., 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. Published by Elsevier Ltd.)

The rare earth element (REE) pollution in the hydrosphere has become a matter of serious concern lately. In this study, using inductively coupled plasma mass spectrometry, the REE contents in soft tissue, byssus and shell liquor of the mussel Mytilus galloprovincialis, as a potential REE pollution bioindicator, in the Black Sea were determined for the first time. The highest REE levels were observed in mussels from the seabed. The REE contents in byssus and shell liquor were higher than in soft tissue. In byssus, the contents of Y, La, Ce, Nd and Dy were the highest, whereas in shell liquor, the heavier REE and Sc were the most concentrated. No likely REE-associated risks from the mussel meat consumption were detected. In soft tissue and byssus, REE contents significantly correlated with that of silicon., 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.)

As filter feeders, marine bivalves inhabiting estuarine and coastal areas are directly exposed to microplastics (MPs) in water. To assess whether MPs number, and their shape, size, colour, and polymer type present in mussels (Mytilus galloprovincialis) and cockles (Cerastoderma edule) varied over one year, bivalves were collected over the year of 2019 in the lower part of the coastal Aveiro lagoon, Portugal. After extraction from the bivalve's whole-body soft tissues, a subset of the visually inspected particles was randomly separated for identification using the Fourier-transform mid-infrared (FT-MIR) spectroscopy. A fraction of the inspected particles, 26-32% of particles >100 μm, and 59-100% of smaller ones were confirmed as MPs. Concentrations varied within the intervals of 0.77-4.3 items g -1 in mussels and 0.83-5.1 items g -1 in cockles, with the lowest values observed in January. In winter, the accumulation of large-sized fibers was composed of a mixture of plastic types, which contrasted against the most abundant MPs in summer consisting mainly of polyethylene of diverse size classes and shapes. Temperature decrease registered in winter might have triggered a lower filtration rate, resulting in lower MPs concentrations in the whole-soft body tissues of organisms. Different properties of MPs found in bivalves between January-February and August-September appear to reflect changes in the characteristics of MPs available in the Aveiro lagoon., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Active biomonitoring of chemical contamination (e.g., Cd, Hg, Pb, DDT, PCB, PAH) in French Mediterranean coastal waters has been performed for more than two decades. This study aimed at presenting the current contamination in 2021 and the temporal evolution of concentrations from 2000. Based on a relative spatial comparison, low concentrations were measured in 2021 at most sites (>83 %). Also, several stations with moderate to high levels were highlighted in the vicinity of major urban industrial centers (e.g., Marseille, Toulon) and near river mouths (e.g., Rhône, Var). Over the last 20 years, no major trend was revealed, mostly, especially for the relative high-level sites. This likely constant contamination over time, plus slight increases of metallic elements at a few sites, still raise questions on the efforts that remain to be made. The decreasing trends of organic compounds, in particular PAH, provide evidence of the efficiency of some management actions., 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.)

In this study, concentrations of elements were determined in edible tissues of Mytilus galloprovincialis by means of inductively coupled plasma mass spectrometry (ICP-MS). The mean levels (mg kg -1 ) of 0.67 for Cd, 6.9 for As, 0.79 for Pb, 2 for Ni, and 42.6 for Zn exceeded the maximum limits in the digestive glands. Also, the mean concentrations of Cd and As in muscle and of Cd and Ni in gills were above the maximum limits. The highest value was found for As in a digestive gland, with 65.4% of the Provisional Tolerable Weekly Intake. In addition, the lowest percentage belonged to Zn with 0.2% of PTWI in muscles and 0.3% of PTWI in gills of the mussels. Mercury concentrations were well below legal limits.

This study evaluated the microplastic abundance, shape, color, size and chemical composition of microplastic in mussels and estimated human exposure to microplastic through consumption of mussels collected from Boka Kotorska Bay (Adriatic coast of Montenegro). Microplastic was found in 53.3% of the studied mussels, with an average microplastic abundance of 2.53 ± 1.1 items/individual. Most of the ingested microplastic were fibers (63.7%), which were blue in color. FT-IR revealed that 98% of the examined particles were plastic, with seven polymers identified, of which polyethylene, polypropylene, and polyethylene terephthalate were the most abumdant polymers in mussels. Three of the polymers detected in mussels (polyamide, polyvinyl chloride and polystyrene) are classified as hazardous by the European Chemical Agency with warning or danger signals. With one serving of mussels, consumers would ingest 22.7 microplastic particles, while the annual dietary intake of microplastic via consumption of mussels was estimated at 99 MP/year., 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.)

Imidazolium ionic liquids (ILs) are chemical compounds beginning to be used on a mass scale. Although their presence in the environment is usually treated as only potential threat, there are already first evidences that this has become a real case. Taking into account their increasing use it might be expected that this problem will also increase in the nearest future. Given that some of the imidazolium cations exhibit high potential for bioconcentration, it is likely that they will accumulate in the tissues of wild organisms. Thus, there is no doubt that monitoring the presence of these compounds in organisms from potentially contaminated waters will be needed. Therefore, the aim of our study was to develop and fully validate a novel and reliable analytical procedure for the determination of the mixture of imidazolium ILs in Mytilus trossulus mussels. For this purpose, different extraction techniques were tested such as: microwave-assisted extraction (MAE), accelerated-solvent extraction (ASE) and bullet-blender homogenization (HOMO). Finally, the proposed procedure is based on the application of MAE technique for the extraction of imidazolium cations and SPE technique using Oasis HLB cartridges for the purification of the obtained extracts and LC-MS/MS technique with QqQ analyzer for their final determination. Absolute recoveries of the proposed analytical procedure reached 71-90%. The developed procedure is characterized with low limits of quantification, at 50-100 ng g -1 dry tissue and allows for reliable determination of trace amounts of the tested compounds in complex biological matrix. Matrix effects obtained for the optimized procedure ranged from 7.8 to 37.5%. As a result, this is the first study presenting the analytical procedure for the analysis of imidazolium ILs in aquatic animal tissues., 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 © 2022. Published by Elsevier B.V.)

The Pacific coast of Canada has a rich marine fauna and a growing human population with increasing potential for pollution releases, but there is currently little overlap between marine wildlife hotspots and ongoing biomonitoring efforts for less bioaccumulative contaminants such as polycyclic aromatic compounds (PAC) and trace metals (metals). We surveyed PACs and metals at marine bird breeding colonies in coastal British Columbia in 2018 by analyzing chemical residues in the soft tissue of bivalve Mytilus sp. mussels collected from stations (n = 3) at seven sites. The concentration of sum PACs (∑ 43 PAC) and high molecular weight (HMW) PACs were highest at the Second Narrows colony in Vancouver Harbour, a highly urbanized and industrialized port within the Salish Sea. For conservation areas, two Salish Sea and three Pacific Ocean coast colonies, PACs were generally lower. However, ∑ 43 PAC, ∑ HMW PAC, and several HMW congeners at the remote site of Triangle Island, a Marine National Wildlife Area, were not significantly different from Second Narrows. The dominant PAC sources at all sites are likely pyrogenic rather than petrogenic, as suggested by PAC profiles, proportion of parent PACs, and source-indicator congeners. For metals, site differences were found for seven out of eight priority metals, but principal component analysis indicated that site differences, such as high mercury and cadmium at offshore sites, are likely related to environmental and biological variables including salinity, condition index, water temperature, and shell length. Our survey across a broad coastal region shows that PAC and metal biomonitoring programs with mussels should include wildlife hotspots where the exposure of protected vertebrate species to pollutants with low bioaccumulation potential would be less obvious, and shows that collection of data on key covariates (e.g. lipid content, salinity) will be critical to tracking long-term trends and detecting pollution release events., 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., (Crown Copyright © 2022. Published by Elsevier Ltd. All rights reserved.)

Mussels, whose soft tissues are used in food and are rich in high-quality protein, are essential in marine fisheries. Mediterranean mussels (Mytilus galloprovincialis Lam) used to assesses the pollution level of a coastal marine zone due to exposure to local technological processes populate the metal piles of a pier built for the 2014 Olympics in Sochi Olympic Park. The chemical composition of mussel soft tissues (61 elements analyzed by the ICP method) indicates anthropogenic pollution of the coastal zone ecosystem, most pronounced in the 50-m zone along the coast due to run off from the terrestrial ecosystem affected by the Olympic construction. Line measurements of annual rings of 6-year-old individuals of mussels showed stabilization of the ecological situation in the coastal-marine zone by 2019. This study can be used for comprehensive ecological monitoring of coastal-marine zones with similar conditions and anthropogenic load., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Due to current marine pollution, microplastics ingestion through seafood is an increasing risk for consumers. In this study, microplastics from mussels (Mytilus galloprovincialis) and table salt employed in popular dishes in Bay of Biscay (Spain) were quantified and analysed by Fourier-Transformed Infrared spectroscopy. Microplastics varied in mussels (mean 0.55-3.20 items/g) depending on the environmental pollution of the collection point (seawater, 0.002-0.015 items/mL; sand, 0.06-0.38 items/g). Microplastics content in table salt (0.1-0.38 items/gr) was much lower than in mussels. Chemical substances found from microplastics in mussels and salt are catalogued as hazardous for human health. Significant correlation between microplastics in sand and mussels was found, suggesting that consumers' risk of microplastics ingestion depends on the harvesting area. Routine microplastics analysis in mussels and disclosure of microplastics content on seafood labels are recommended for conscious, informed consumption., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Mussels have a particular nutritional value, representing a highly valued food source and thus sought after worldwide. Their meat is a real culinary delicacy, rich in proteins, lipids, carbohydrates, trace elements, enzymes, and vitamins. The seasonal variation of mussels' biochemical composition has been studied to determine the best harvesting period to capitalize on various biologically active fractions. In this work biochemical determinations have been performed on fresh flesh samples of Mytilus galloprovincialis specimens from the Black Sea coast to study seasonal variations in mussels' biochemical compounds. An analysis of significant lipid classes and the fatty acid composition of lipid extracts obtained from mussel flesh has also been performed. Since mussels retain pollutants from the marine environment, in parallel, the concentration of heavy metals in the meat of mussels collected for the analysis of the chemical composition was investigated. The impact and risk of heavy metal poisoning due to food consumption of mussels contaminated due to pollution of the marine harvesting area was evaluated by the bio-concentration factor of metals and estimated daily intakes of heavy metals through mussel consumption.

Paralytic shellfish toxins (PSTs) are produced mainly by Alexandrium catenella (formerly A. tamarense ). Since 2000, the National Institute of Fisheries Science (NIFS) has been providing information on PST outbreaks in Korean coastal waters at one- or two-week intervals. However, a daily forecast is essential for immediate responses to PST outbreaks. This study aimed to predict the outbreak timing of PSTs in the mussel Mytilus galloprovincialis in Jinhae Bay and along the Geoje coast in the southern coast of the Korea Peninsula. We used a long-short-term memory (LSTM) neural network model for temporal prediction of PST outbreaks from environmental data, such as water temperature (WT), tidal height, and salinity, measured at the Geojedo, Gadeokdo, and Masan tidal stations from 2006 to 2020. We found that PST outbreaks is gradually accelerated during the three years from 2018 to 2020. Because the in-situ environmental measurements had many missing data throughout the time span, we applied LSTM for gap-filling of the environmental measurements. We trained and tested the LSTM models with different combinations of environmental factors and the ground truth timing data of PST outbreaks for 5479 days as input and output. The LSTM model trained from only WT had the highest accuracy (0.9) and lowest false-alarm rate. The LSTM-based temporal prediction model may be useful as a monitoring system of PSP outbreaks in the coastal waters of southern Korean.

Tetrodotoxins (TTXs), potent neurotoxins, have become an increasing concern in Europe in recent decades, especially because of their presence in mollusks. The European Food Safety Authority published a Scientific Opinion setting a recommended threshold for TTX in mollusks of 44 µg equivalent kg -1 and calling all member states to contribute to an effort to gather data in order to produce a more exhaustive risk assessment. The objective of this work was to assess TTX levels in wild and farmed mussels ( Mytilus galloprovincialis ) harvested in 2018-2019 along the coastal area of the Marche region in the Central Adriatic Sea (Italy). The presence of Vibrio spp. carrying the non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) genes, which are suspected to be involved in TTX biosynthesis, was also investigated. Out of 158 mussel samples analyzed by hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry (HILIC-MS/MS), 11 (7%) contained the toxins at detectable levels (8-26 µg kg -1 ) and 3 (2%) contained levels above the EFSA safety threshold (61-76 µg kg -1 ). Contaminated mussels were all harvested from natural beds in spring or summer. Of the 2019 samples, 70% of them contained V. alginolyticus strains with the NRPS and/or PKS genes. None of the strains containing NRPS and/or PKS genes showed detectable levels of TTXs. TTXs in mussels are not yet a threat in the Marche region nor in Europe, but further investigations are surely needed.

The international seafood trade is based on food safety, quality, sustainability, and traceability. Mussels are bio-accumulative sessile organisms that need regular control to guarantee their safe consumption. However, no well-established and validated methods exist to trace mussel origin, even if several attempts have been made over the years. Recently, an inorganic multi-elemental fingerprint coupled to multivariate statistics has increasingly been applied in food quality control. The mussel shell can be an excellent reservoir of foreign inorganic chemical species, allowing recording long-term environmental changes. The present work investigates the multi-elemental composition of mussel shells, including Al, Cu, Cr, Zn, Mn, Cd, Co, U, Ba, Ni, Pb, Mg, Sr, and Ca, determined by inductively-coupled plasma mass-spectrometry in Mytilus galloprovincialis collected along the Central Adriatic Coast (Marche Region, Italy) at 25 different sampling sites (18 farms and 7 natural banks) located in seven areas. The experimental data, coupled with chemometric approaches (principal components analysis and linear discriminant analysis), were used to create a statistical model able to discriminate samples as a function of their production site. The LDA model is suitable for achieving a correct assignment of >90% of individuals sampled to their respective harvesting locations and for being applied to counteract fraud.

Anthropogenic warming and ocean acidification are predicted to negatively affect marine calcifiers. While negative effects of these stressors on physiology and shell calcification have been documented in many species, their effects on shell mineralogical composition remains poorly known, especially over longer time periods. Here, we quantify changes in the shell mineralogy of a foundation species, Mytilus californianus , under 60 y of ocean warming and acidification. Using historical data as a baseline and a resampling of present-day populations, we document a substantial increase in shell calcite and decrease in aragonite. These results indicate that ocean pH and saturation state, not temperature or salinity, play a strong role in mediating the shell mineralogy of this species and reveal long-term changes in this trait under ocean acidification., Competing Interests: The authors declare no competing interest.

Carotenoids consist of a series of conjugated isoprene units that are characteristically highly conjugated through double bonds, leading to the formation of many isomers that are susceptible to oxidation and other chemical modifications. Extreme hydrophobicity and high complexity make carotenoids difficult to identify and quantify. We implemented the use of a common Syncronis C18 column with strong eluting solvent, here isopropanol, to successfully separate a mixture of 23 carotenoids standards with different structural properties. In addition, the method differentiated between three groups of isomeric carotenoids (lycopene/δ-carotene/γ-carotene/ε-carotene/α-carotene/β-carotene, α-cryptoxanthin/β-cryptoxanthin, and zeaxanthin/lutein) by optimizing the gradient profile and using liquid chrmatography-mass spectrometry. The LOD ranged from 0.05 to 5.51 ng/mL, and the recovery of carotenoids in Mytilus coruscus was from 63.54 to 93.25%, with standard deviations <10%. Twenty-five carotenoids were detected with a total content of 857 ± 55.1 mg/kg, and three isomeric carotenoids were identified: ε-carotene, α-carotene, and β-carotene. Our results show that this methodology is a significant improvement over other alternatives for analyzing carotenoids because of its compatibility with carotenoids of different categories, and most importantly, its ability to resolve isomeric carotenes, which is significant not only for assessing carotenoid species, but also for the tracing of metabolic pathways of carotenoids., (© 2020 Wiley-VCH GmbH.)

Plastics profoundly threatens ecological balance in marine ecosystems across the globe in the current era of industrialization. Microplastics (MP), in particular, can pose risks reaching humans through the food web via various marine organisms. Among these organisms, since they are consumed as a whole, mussels are vital vectors of MP transfer during human consumption. Hence, here we analyzed MP pollution in Mediterranean mussel (Mytilus galloprovincialis) sampled from 23 different locations all along the Turkish coasts of the Black Sea, Sea of Marmara, and the Aegean Sea. After digestion of the mussels with H 2 O 2 , the micro-particles were determined under a stereomicroscope and characterized by confirming with FTIR analyses. 48% of the sampled mussels were found to have MPs. The average MP abundance was 0.69 item/mussel and 0.23 item/g fresh weight (fw) of soft tissue. Morphology was ordered as follows: fragments (67.6%)> fibers (28.4%)> films (4.05%). The dominant size of MPs was detected less than 0.5 mm (26.58%). 12 different polymers have been identified by FTIR and PET (32.9%), PP (28.4%), and PE (19.4%) were found to constitute 80% of the total MPs. The annual average exposure amount for mussel consumers in Turkey was estimated as 1918 MPs item/per year. Even though international organizations such as FAO, JECFA, or EU have not declared permissible limits, our data may inform human health uptake of MP ingestion via mussels. This data might also serve as a reference data-set for further MP monitoring research in Turkish and European Seas., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Graphene oxide (GO)-based composite materials have become widely popular in many applications due to the attractive properties of GO, such as high strength and high electrical conductivity at the nanoscale. Most current GO composites use organic polymer as the matrix material and thus, their synthesis suffers from the use of organic solvents or surfactants, which raise environmental and energy-consumption concerns. Inspired by mussel foot proteins (Mfp) secreted by the saltwater mussel, Mytilus galloprovincialis and by recent advances in microbial protein production, we developed an aqueous-based green synthesis strategy for preparing GO/Mfp film composites. These GO/Mfp films display high tensile strength (134-158 MPa), stretchability (~ 26% elongation), and high toughness (20-24 MJ/m 3 ), beyond the capabilities of many existing GO composites. Renewable production of Mfp proteins and the facile fabrication process described provides a new avenue for composite material synthesis, while the unique combination of mechanical properties of GO/Mfp films will be attractive for a range of applications.

Seafood represents a significant part of the human staple diet. In the recent years, the identification of emerging lipophilic marine toxins has increased, leading to the potential for consumers to be intoxicated by these toxins. In the present work, we investigate the presence of lipophilic marine toxins (both regulated and emerging) in commercial seafood products from non-European locations, including mussels Mytilus chilensis from Chile, clams Tawerea gayi and Metetrix lyrate from the Southeast Pacific and Vietnam, and food supplements based on mussels formulations of Perna canaliculus from New Zealand. All these products were purchased from European Union markets and they were analyzed by UPLC-MS/MS. Results showed the presence of the emerging pinnatoxin-G in mussels Mytilus chilensis at levels up to 5.2 µg/kg and azaspiracid-2 and pectenotoxin-2 in clams Tawera gayi up to 4.33 µg/kg and 10.88 µg/kg, respectively. This study confirms the presence of pinnatoxins in Chile, one of the major mussel producers worldwide. Chromatograms showed the presence of 13-desmethyl spirolide C in dietary supplements in the range of 33.2-97.9 µg/kg after an extraction with water and methanol from 0.39 g of the green lipped mussels powder. As far as we know, this constitutes the first time that an emerging cyclic imine toxin in dietary supplements is reported. Identifying new matrix, locations, and understanding emerging toxin distribution area are important for preventing the risks of spreading and contamination linked to these compounds.

Ocean acidification alters physiology, acid-base balance and metabolic activity in marine animals. Near future elevated pCO 2 conditions could be expected to influence the bioaccumulation of metals, feeding rate and immune parameters in marine mussels. To better understand such impairments, a series of laboratory-controlled experiment was conducted by using a model marine mussel, Mytilus galloprovincialis. The mussels were exposed to three pH conditions according to the projected CO 2 emissions in the near future (one ambient: 8.10 and two reduced: 7.80 and 7.50). At first, the bioconcentration of Ag and Cd was studied in both juvenile (2.5 cm) and adult (5.1 cm) mussels by using a highly sensitive radiotracer method ( 110m Ag and 109 Cd). The uptake and depuration kinetics were followed 21 and 30 days, respectively. The biokinetic experiments demonstrated that the effect of ocean acidification on bioconcentration was metal-specific and size-specific. The uptake, depuration and tissue distribution of 110m Ag were not affected by elevated pCO 2 in both juvenile and adult mussels, whereas 109 Cd uptake significantly increased with decreasing pH in juveniles but not in adults. Regardless of pH, 110m Ag accumulated more efficiently in juvenile mussels than adult mussels. After executing the biokinetic experiment, the perturbation was sustained by using the same mussels and the same experimental set-up, which enabled us to determine filtration rate, haemocyte viability, lysosomal membrane stability, circulating cell-free nucleic acids (ccf-NAs) and protein (ccf-protein) levels. The filtration rate and haemocyte viability gradually decreased by increasing pCO 2 level, whereas the lysosomal membrane stability, ccf-NAs, and ccf-protein levels remained unchanged in the mussels exposed to elevated pCO 2 for eighty-two days. This study suggests that acidified seawater partially shift metal bioaccumulation, physiological and cellular parameters in the mussel Mytilus galloprovincialis., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Mollusk shell is a product of biomineralization with excellent mechanical properties, and the shell matrix proteins (SMPs) have important functions in shell formation. A vWA domain-containing protein (VDCP) was identified from the shell of Mytilus coruscus as a novel shell matrix protein. The VDCP gene is expressed at a high level in specific locations in the mantle and adductor muscle. Recombinant VDCP (rVDCP) showed abilities to alter the morphology of both calcite and aragonite, induce the polymorph change of calcite, bind calcite, and decrease the crystallization rate of calcite. In addition, immunohistochemistry analyses revealed the specific location of VDCP in the mantle, the adductor muscle, and the myostracum layer of the shell. Furthermore, a pull-down analysis revealed eight protein interaction partners of VDCP in shell matrices and provided a possible protein-protein interaction network of VDCP in the shell.

Paralytic shellfish toxins (PSTs) produced by Alexandrium catenella (formerly A. tamarense ) in Korean coastal waters caused the deaths of four people (in 1986 and 1996) who consumed contaminated mussels ( Mytilus edulis ). This led to more detailed consideration of the risks of PST outbreaks and incidents in Korea, including the introduction of shellfish collection bans. In this study, we investigated the relationships between A. catenella population dynamics and PST accumulation in the mussel M. galloprovincialis. Discharges from the Nakdong River affect the environmental conditions along the Geoje coast, resulting in low salinity and high nutrient levels that trigger blooms of A. catenella . At the toxin peak on 24 April 2017, the toxins detected in A. catenella cells were C1, gonyautoxin (GTX)1 and GTX2, whereas the concentrations of PSTs in M. galloprovincialis were high and in the order of GTX4 > GTX1 > GTX3 > saxitoxin (STX) > GTX2 > neoSTX > decarbamoylgonyautoxin (dcGTX)2 > dc GTX3. The PST level in mussels was also high. At 15 °C, the PSTs are constantly found to be higher (10-fold higher in 2017 and 30-fold higher in 2018) than safe levels for human consumption (80 μg STX diHCl equivalents 100 g -1 ).

Lipophilic marine toxins (LMTs) are made up of multiple groups of toxic analogues, which are characterised by different levels of cellular and toxic action. The most prevalent groups in the southern Pacific zone are: a) okadaic acid group (OA-group) which consists of okadaic acid (OA) and dinophysistoxin-1 (DTX-1); and, b) pectenotoxin-2 (PTX2) group which consists of pectenotoxin-2 (PTX-2). The main objective of our study was to examine in vitro biotransformation of OA-group and PTX-group in the tissues of two endemic species of bivalves from southern Chile; blue mussels ( Mytilus chilensis ) and clams ( Ameghinomya antiqua ). The biotransformation processes of both groups were only detected in the digestive glands of both species using LC-MS/MS. The most frequently detected analogues were acyl derivatives (≈2.0 ± 0.1 μg ml -1 ) for OA-group and PTX-2SA (≈1.4 ± 0.1 μg ml -1 ) for PTX-group, with a higher percentage of biotransformation for OA-group ( p < .001). In addition, simultaneous incubations of the different analogues (OA/PTX-2; DTX-1/PTX-2 and OA/DTX-1/PTX-2) did not show any interaction between the biotransformation processes. These results show that the toxicological variability of endemic species leads to biotransformation of the profile of toxins, so that these new analogues may affect people's health.

Cyclic imines (CIs) are emerging marine lipophilic toxins (MLTs) occurring in microalgae and shellfish worldwide. The present research aimed to study CIs in mussels farmed in the Adriatic Sea (Italy) during the period 2014-2015. Twenty-eight different compounds belonging to spirolides (SPXs), gymnodimines (GYMs), pinnatoxins (PnTXs) and pteriatoxins (PtTXs) were analyzed by the official method for MLTs in 139 mussel samples collected along the Marche coast. Compounds including 13-desmethyl spirolide C (13-desMe SPX C) and 13,19-didesmethyl spirolide C (13,19-didesMe SPX C) were detected in 86% of the samples. The highest levels were generally reported in the first half of the year reaching 29.2 µg kg -1 in January/March with a decreasing trend until June. GYM A, for the first time reported in Italian mussels, was found in 84% of the samples, reaching the highest concentration in summer (12.1 µg kg -1 ). GYM A and SPXs, submitted to tissue distribution studies, showed the tendency to accumulate mostly in mussel digestive glands. Even if SPX levels in mussels were largely below the European Food Safety Authority (EFSA) reference of 400 μg SPXs kg -1 , most of the samples contained CIs for the large part of the year. Since chronic toxicity data are still missing, monitoring is surely recommended.

Microplastic (MP) contamination in marine organisms is a growing field of research internationally. However not much is known about MP presence in invertebrates in southern Africa. The aim of this study was to determine whether MPs occurs in mussels prevalent in Cape Town, South Africa. Mussels (Mytilus galloprovincialis, Choromytilus meridionalis and Aulacomya ater) were sampled at 27 sites in October 2018 and MPs were recorded in 98% of mussels analysed. Most MPs were filaments, dark in colour and the size ranged between 50 and 1000 µm. There were no significant differences in MPs between the mussel species, with an average of 2.33 (standard error ± 0.2) MP particles/g and 4.27 (standard error ± 0.5) particles/individual being recorded for all sites combined. This is the first record of MPs in mussels in the region and provides a baseline for further investigations and monitoring of MPs.

Bivalve mollusks as filter-feeders concentrate in their digestive tissue microorganisms likely present in the harvesting water, thus becoming risky food especially if consumed raw or poorly cooked. To eliminate bacteria and viruses eventually accumulated, they must undergo a depuration process which efficacy on viruses is on debate. To better clarify the worth of the depuration process on virus elimination from mussels, in this study we investigated rotavirus kinetics of accumulation and depuration in Mytilus galloprovincialis experimentally contaminated. Depuration process was monitored for 9 days and virus residual presence and infectivity were evaluated by real time quantitative polymerase chain reaction, cell culture and electron microscopy at days 1, 2, 3, 5, 7, 9 of depuration. Variables like presence of ozone and of microalgae feeding were also analyzed as possible depuration enhancers. Results showed a two-phase virus removal kinetic with a high decrease in the first 24 h of depuration and 5 days necessary to completely remove rotavirus.

This study presents the development and validation of a new analytical method based on matrix solid-phase dispersion (MSPD), integrating sample extraction and clean-up in one single step, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the simultaneous determination of 18 organophosphorus flame retardants and/or plasticizers (OPEs) in marine mussel (Mytilus edulis and Mytilus galloprovincialis) samples. Among these OPEs, 5 (tetraethyl 1,2-ethanediylbis(phosphonate), 6H-dibenzo[c,e] [1,2]oxaphosphinine 6-oxide, tris(2,3-dibromopropyl) phosphate, 2,2-propanediyldi-4,1-phenylene bis(phosphate) and resorcinol bis(diphenyl phosphate)) are considered here for the first time in marine samples. Different parameters affecting the MSPD (clean-up sorbent and elution solvent) were optimized to obtain a good compromise between analyte recoveries and extract clean-up. Also, particular attention was paid to tackle blank issues. The overall method was validated in terms of trueness, precision and detection and quantification limits. Percentages of recovery varied from 69% to 122% with relative standard deviations below 24%. Detection limits ranged from 0.06 to 5 ng g -1 and quantification limits from 0.19 to 17 ng g -1 dry weight. Finally, the method was applied to the analysis of 7 mussel samples collected in the coast of Galicia (Spain). 8 OPEs were detected in these samples at concentrations ranging from the LOQ to 291 ng g -1 dry weight., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The purpose of this paper is to investigate the trace metal content in edible biomonitors (i.e., mollusks) in the Beagle Channel (southern Patagonia) and to assess the human health risks associated with their consumption. Rationale: The monitoring breakdown structure (MBS) conceptual model was applied to four sampling campaigns (2005 → 2012) that collected 729 samples of Mytilus chilensis and Nacella magellanica. The composition of trace elements (Cd, Cr, Cu, Ni, Pb and Zn) in the mollusks was determined using graphite furnace (GFAAS) or flame atomic absorption spectrometry (FAAS). We compared the mean obtained values with the maximum levels (MLs) of each element established by international organizations. Then, based on semi-structured interviews, we calculated the estimated daily intake (EDI) of local residents and compared it with safety reference doses, i.e., the provisional tolerable daily intake (PTDI), provisional maximum tolerable daily intake (PMTDI), and tolerable daily intake (TDI), as well as the benchmark dose level lower confidence limit for Pb (BMDL 01 , a reference point (RP)/point of departure (POD). Moreover, to obtain information about the potential health risks of ingesting heavy metals (HMs) through mollusk consumption, we evaluated the target hazard quotient (THQ) and the hazard index (HI). Findings: For Cd and Pb, 65% and 40% of bivalves exceeded the MLs established by the Mercado Común del Sur (Mercosur), respectively. Except for Cd in N. magellanica (i.e., 1.20 μg/kg/bw/day), EDI values were clearly lower than the safety reference doses. For Cr, Cu, Ni, Pb and Zn, mussels were safe for consumption and did not raise concerns for public health. Likewise, THQ values were well below one for most of the studied metals, indicating that the exposed human population is assumed to be safe. Occasional high consumers of mollusks from the most contaminated sites may be at some health risk. Originality: The food production system and the environment are complex systems; this is crucial to understand when we consider ecosystems as a food source (i.e., marine ecosystems). Here we consider edible biomonitors, that are organisms that can have a dual function. They are food, and at the same time, if properly calibrated, they can act as indicators of environmental quality. This study is the first to investigate relevant essential and non-essential trace metal content in two edible mollusks from the Beagle Channel in a long-term survey (2005 → 2012). The information variety was high; approximately thirteen thousand determinations were conducted to support the risk assessment for mollusk consumption. Other aspects connected with the health risks and the uncertainty factors related to the presence of essential and non-essential minerals in edible mollusks as well as the use of the MBS are also discussed., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

We assessed the relationship between V, Cr, Mn, Hg, As, Cd, Sn, Sb and Pb concentrations in Mytilus galloprovincialis samples from the coasts of Sicily and the expression of metallothioneins . Toxic mineral elements assessment was carried out by A.A. Spectrometry and ICP-MS. The metallothioneins expression was performed by q-PCR method. Low metals' levels were found in the mussel samples examined, in comparison with what was reported in literature. The highest mean values of toxic mineral elements were found in Gela (Cr 0.178 ± 0.03 mg/Kg, Mn 4.325 ± 0.012 mg/Kg, As 3.706 ± 0.009 mg/Kg, Sn 0.148 ± 0.014 mg/Kg, Sb 0.009 ± 0.004 mg/Kg e Pb 0.364 ± 0.01 mg/Kg). Significant levels of Hg were found in samples from Catania (0.014 ± 0.005 mg/Kg). Only vanadium and lead concentrations showed significant differences between sampling areas ( p  < 0.05). Molecular analysis verified a basal expression of Mt1 and the absence of over-expression of Mt2 , confirming the low mineral's concentrations found in the samples examined.

This study aimed to obtain the purified fractions of Mytilus coruscus polysaccharides (MCPs) and investigate their antioxidant activities. MCPs were prepared through ultrasonic-assisted enzymatic extraction optimized by employing the response surface methodology. A single-factor experiment was conducted using the Box-Behnken design to determine the optimum extraction conditions of MCPs. The ultrasonic power was 60 W, liquid-to-material ratio was 30 mL/g, extraction time was 36 min, extraction temperature was 64 °C, enzyme concentration was 3.2%, and polysaccharide extraction yield was 12.86% ± 0.12%. A novel polysaccharide (MCP1-2) was obtained after the purification with AB-8 macroporous resin, DEAE Sepharose Fast Flow, and Sepharose CL-6B column. The molecular weight of MCP1-2 was estimated to be 134.9 kDa according to high-performance gel permeation chromatography. High-pressure liquid-phase chromatography results showed that MCP1-2 contained mannose, rhamnose, glucuronic acid, glucose, galactose, and L-Fuc at a molar ratio of 1.53:1:4.83:81.82:2.36:1.51. Infrared and NMR spectroscopies confirmed that MCP1-2 possessed α- and β- configurations. The antioxidant activities of MCP1-2 were investigated in vitro, and the results showed that MCP1-2 had good antioxidant activity and can be used as a natural antioxidant in food., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The present study was designed to evaluate in vivo the oral bioavailability of lead (Pb) present in the marine bivalve Dosinia exoleta. This infaunal clam, despite inhabiting in clean areas, presents Pb concentrations that are over the 1.5 mg kg -1 wet weight limit for human consumption set by the European Commission. However, Pb is accumulated in this clam in the form of metal rich granules, and it has been shown to be unavailable for trophic transfer to a marine decapod, so it was hypothesised that it might be unavailable for human consumers as well. Twelve Sprague Dawley rats were fed during 14 days with a diet including control mussels (Mytilus galloprovincialis), D. exoleta, or mussels enriched in Pb to the same levels as those found in D. exoleta. Pb accumulation in different rat tissues (blood, bone, kidneys and liver) was analysed. It was observed that Pb assimilation from D. exoleta was about half of Pb assimilation from M. galloprovincialis, and absolute bioavailabilities were around 2% for M. galloprovincialis and 1% for D. exoleta. These results suggest that it might be possible to increase the limit for human consumption for this bivalve to 3 mg kg -1 wet weight without representing an increase in the risk for consumers., (Copyright © 2019 Elsevier Inc. All rights reserved.)

This study investigated the role of a municipal wastewater treatment plant (WWTP) effluent and an abandoned coastal landfill as pathways for microplastics (MPs) input into the marine environment. MPs were first analyzed in raw sewage influent, sludge and effluent samples, and their fate was studied along a distance gradient from the WWTP in three matrices: surface water, sediments and wild mussels. All suspected MPs were characterized according to their polymer nature using micro-Raman spectroscopy. The investigated WWTP had an estimated daily discharge of 227 million MPs. MPs were found in all matrices with a decreasing abundance from the effluent. Strong MPs abundances (higher than those found near the WWTP effluent) were observed in the vicinity of the coastal landfill suggesting its importance as a MPs entry route into the marine coastal environment. Our study supports the idea that blue mussels are a promising sentinel species for MPs (<200 μm)., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

MytiLec-1, a 17 kDa lectin with β-trefoil folding that was isolated from the Mediterranean mussel ( Mytilus galloprovincialis ) bound to the disaccharide melibiose, Galα(1,6) Glc, and the trisaccharide globotriose, Galα(1,4) Galβ(1,4) Glc. Toxicity of the lectin was found to be low with an LC 50 value of 384.53 μg/mL, determined using the Artemia nauplii lethality assay. A fluorescence assay was carried out to evaluate the glycan-dependent binding of MytiLec-1 to Artemia nauplii. The lectin strongly agglutinated Ehrlich ascites carcinoma (EAC) cells cultured in vivo in Swiss albino mice. When injected intraperitoneally to the mice at doses of 1.0 mg/kg/day and 2.0 mg/kg/day for five consecutive days, MytiLec-1 inhibited 27.62% and 48.57% of cancer cell growth, respectively. Antiproliferative activity of the lectin against U937 and HeLa cells was studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in vitro in RPMI-1640 medium. MytiLec-1 internalized into U937 cells and 50 μg/mL of the lectin inhibited their growth of to 62.70% whereas 53.59% cell growth inhibition was observed against EAC cells when incubated for 24 h. Cell morphological study and expression of apoptosis-related genes (p53, Bax, Bcl-X, and NF-κB) showed that the lectin possibly triggered apoptosis in these cells.

Although by now the glass transition temperature of uncrystallized bulk water is generally accepted to manifest at temperature T g near 136 K, not much known are the spectral dispersion of the structural α-relaxation and the temperature dependence of its relaxation time τ α,bulk (T). Whether bulk water has the supposedly ubiquitous Johari-Goldstein (JG) β-relaxation is a question that has not been answered. By studying the structural α-relaxation over a wide range of temperatures in several aqueous mixtures without crystallization and with glass transition temperatures T g close to 136 K, we deduce the properties of the α-relaxation and the temperature dependence of τ α,bulk (T) of bulk water. The frequency dispersion of the α-relaxation is narrow, indicating that it is weakly cooperative. A single Vogel-Fulcher-Tammann (VFT) temperature dependence can describe the data of τ α,bulk (T) at low temperatures as well as at high temperatures from neutron scattering and GHz-THz dielectric relaxation, and hence, there is no fragile to strong transition. The T g -scaled VFT temperature dependence of τ α,bulk (T) has a small fragility index m less than 44, indicating that water is a "strong" glass-former. The existence of the JG β-relaxation in bulk water is supported by its equivalent relaxation observed in water confined in spaces with lengths of nanometer scale and having Arrhenius T-dependence of its relaxation times τ conf (T). The equivalence is justified by the drastic reduction of cooperativity of the α-relaxation in nanoconfinement and rendering it to become the JG β-relaxation. Thus, the τ conf (T) from experiments can be taken as τ β,bulk (T), the JG β-relaxation time of bulk water. The ratio τ α,bulk (T g )/τ β,bulk (T g ) is smaller than most glass-formers, and it corresponds to the Kohlrausch α-correlation function, exp[-(t/τ α,bulk ) 1-n ], having (1-n) = 0.90. The dielectric data of many aqueous mixtures and hydrated biomolecules with T g higher than that of water show the presence of a secondary ν-relaxation from the water component. The ν-relaxation is strongly connected to the α-relaxation in properties, and hence, it belongs to the special class of secondary relaxations in glass-forming systems. Typically, its relaxation time τ ν (T) is longer than τ β,bulk (T), but τ ν (T) becomes about the same as τ β,bulk (T) at sufficiently high water content. However, τ ν (T) does not become shorter than τ β,bulk (T). Thus, τ β,bulk (T) is the lower bound of τ ν (T) for all aqueous mixtures and hydrated biomolecules. Moreover, it is τ β,bulk (T) but not τ α (T) that is responsible for the dynamic transition of hydrated globular proteins.

The occurrence of marine harmful algae is increasing worldwide and, therefore, the accumulation of lipophilic marine toxins from harmful phytoplankton represents a food safety threat in the shellfish industry. Galicia, which is a commercially important EU producer of edible bivalve mollusk have been subjected to recurring cases of mussel farm closures, in the last decades. This work aimed to study the toxic profile of commercial mussels ( Mytilus galloprovincialis ) in order to establish a potential risk when ingested. For this, a total of 41 samples of mussels farmed in 3 Rías (Ares-Sada, Arousa, and Pontevedra) and purchased in 5 local markets were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). Chromatograms showed the presence of okadaic acid (OA), dinophysistoxin-2 (DTX-2), pectenotoxin-2 (PTX-2), azaspiracid-2 (AZA-2), and the emerging toxins 13-desmethyl spirolide C (SPX-13), and pinnatoxin-G (PnTX-G). Quantification of each toxin was determined using their own standard calibration in the range 0.1%-50 ng/mL (R2 > 0.99) and by considering the toxin recovery (62-110%) and the matrix correction (33-211%). Data showed that OA and DTX-2 (especially in the form of esters) are the main risk in Galician mollusks, which was detected in 38 samples (93%) and 3 of them exceeded the legal limit (160 µg/kg), followed by SPX-13 that was detected in 19 samples (46%) in quantities of up to 28.9 µg/kg. Analysis from PTX-2, AZA-2, and PnTX-G showed smaller amounts. Fifteen samples (37%) were positive for PTX-2 (0.7-2.9 µg/kg), 12 samples (29%) for AZA-2 (0.1-1.8 µg/kg), and PnTX-G was detected in 5 mussel samples (12%) (0.4 µg/kg-0.9 µg/kg). This is the first time Galician mollusk was contaminated with PnTX-G. Despite results indicating that this toxin was not a potential risk through the mussel ingestion, it should be considered in the shellfish safety monitoring programs through the LC-MS/MS methods.

The influence of sedimentary metals on the cockle A. trapezia tissue was examined using a strong difference in sedimentary metal concentrations in an embayment (Hen and Chicken Bay) highly contaminated in Cu and an adjacent cove (Iron Cove), strongly enriched in Cd, Cu, Pb and Zn within the heavily-urbanised Sydney estuary catchment (Australia). Statistically significant differences were recorded for cockle tissue metal concentrations between the study locations reflecting differences in surficial sediment metal concentrations. Low metal uptake was apparent in A. trapezia tissue, which were considerably less than background sedimentary concentrations and was of low-risk for human consumption. Dissimilar bioconcentration of Cd, Cu, Pb and Zn was apparent in tissue of cockles (A. trapezia), prawns (Metapenaeus bennettae), mussels (Mytilus galloprovincialis) and oysters (Saccostrea glomerata), due possibly to different feeding patterns and biogeochemical conditions in bottom sediments., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The incorporation of silver nanoparticles (AgNPs) in commercial products is increasing rapidly. The consequent release of AgNPs into domestic and industrial wastewater raises environmental concerns due to their anti-microbial properties and toxicity to non-target aquatic organisms. The aim of the present study was to investigate the effects of nanArgen™ (Nanotek S.A.), a AgNP-enabled consumer product, in the marine bivalve Mytilus galloprovincialis. Two environmentally relevant concentrations of nanArgen™ (1 and 10 μg/L) were tested in vivo for 96 h, and Ag was quantified in mussel soft tissue and natural seawater (NSW). nanArgen™ suspensions were characterized via TEM, SEM, EDS, DLS, and UV-vis optical analysis. Several molecular and biochemical responses were investigated in exposed mussels: lysosomal membrane stability by Neutral Red Retention Time (NRRT) assay; micronucleus (MN) frequency in hemocytes; metallothionein (MT) protein content and gene expression (mt10 and mt20); catalase (CAT) and glutathione-S-transferase (GST) activities; malondialdehyde (MDA) accumulation in digestive glands; and efflux activity of ATP-binding cassette transport proteins (ABC) in gill biopsies. SEM, TEM and DLS analyses confirmed the presence of well-defined AgNPs in nanArgen™ which were roughly spherical with an average particle size of approx. 30 ± 10 nm. DLS analysis revealed the formation of AgNP aggregates in nanArgen™ suspension in NSW (Z-average of 547.80 ± 90.23 nm; PDI of 0.044). A significant concentration-dependent accumulation of Ag was found in mussels' whole soft tissue in agreement with a concentration-dependent decrease in NRRT and an increase of MN frequency in hemocytes and GST activities in digestive glands. A significant increase in MDA levels and MT via both molecular and biochemical tests, were also observed but only at the highest nanArgen™ concentration (10 μg/L). No changes were observed in CAT activities. ABC efflux activities in gill biopsies showed a significant decrease (p <  0.05) only at the lowest concentration (1 μg/L). On such basis, nanArgen™ is shown to be able to induce toxicity and Ag accumulation in marine mussels similarly to AgNPs and in short-term exposure conditions at environmentally relevant concentrations. AgNP-enabled products, instead of pristine AgNPs, should be the focus of future ecotoxicity studies in order to address any risks associated to their widespread use, disposal and uncontrolled release into the aquatic environment for non target species., (Copyright © 2019 Elsevier B.V. All rights reserved.)