Chemical and microlitter (ML) pollution in three Estonian coastal areas (Baltic Sea) was investigated using mussels (Mytilus trossulus). Polycyclic aromatic hydrocarbons (PAH) in mussel tissues were observed in moderate levels with high bioaccumulation factors for the more hydrophilic and low molecular weight PAH (LMW PAH), namely anthracene and fluorene. Tissue concentrations of polybrominated diphenyl ethers (PBDE) and cadmium within mussel populations exceeded the Good Environmental Status thresholds by more than 200% and 60%, respectively. Multiple contamination at the Muuga Harbour site by tributyltin, high molecular weight PAH, including the highly toxic benzo[c]fluorene and PBDE, coincided with the inhibition of acetylcholinesterase activity and a lower condition index of the mussels. The metabolization and removal of bioaccumulated LMW PAH, reflected in the dominance of oxy-PAH such as anthracene-9,10-dione, is likely associated with the increased activity of glutathione S-transferase in caged mussels. Only a few microplastic particles were observed among the ML in mussel tissues, with coloured cellulose-based microfibers being the most prevalent. The average concentration of ML in mussels was significantly higher at the harbour area than at other sites. The integrated biomarker response index values allowed for the differentiation of pollution levels across studied locations representing high, intermediate, and low pollution levels within the studied area.

Marine ecosystems are under escalating threats from myriad environmental stressors, necessitating a deeper understanding of their impact on biodiversity and the health of sentinel organisms. In this study, we carried out a spatiotemporal multi-omic analysis of liquid biopsies collected from mussels (Mytilus spp.) in marine ecosystems of a national park. We delved into the epigenomic, transcriptomic, glycomic, proteomic, and microbiomic profiles to unravel the intricate interplay between ecosystem biodiversity and mussels\' biological response to their environments. Our analysis revealed temporal fluctuations in the alpha diversity of the circulating microbiome associated with human activities. Analysis of the hemolymphatic circulating cell-free DNA (ccfDNA) provided information on the biodiversity and the presence of potential pathogens. Epigenomic analysis revealed widespread hypomethylation sites within the mitochondrial (mtDNA). Comparative transcriptomic and glycomic analyses highlighted differences in metabolic pathways and genes associated with immune and wound healing functions. This study demonstrates the potential of multi-omic analysis of liquid biopsy in sentinel to provide a holistic view of human activities\' environmental impacts on marine coastal ecosystems. Overall, this approach has the potential to enhance the effectiveness and efficiency of various conservation efforts, leading to more informed decision-making and better outcomes for biodiversity and ecosystem conservation.

As the demand for rare earth elements (REEs) continues to surge in diverse industrial and medical domains, the ecological consequences of their ubiquitous presence have garnered heightened attention. Among the REEs, gadolinium (Gd), commonly used in medical imaging contrast agents, has emerged as a pivotal concern due to its inadvertent introduction into marine ecosystems via wastewater release. This study delves into the complex ecotoxicological implications of Gd contamination, focusing on its impact on the embryonic development and sperm functionality of Mytilus galloprovincialis. The findings from this study underscore the potential hazards posed by this rare element, offering a critical perspective on the ecological risks associated with Gd. Notably, this exploratory work reveals that Gd exerts a significant embryotoxic effect at elevated concentrations, with an observed half maximal effective concentration (EC50) value of 0.026 mg/L. Additionally, Gd exposure leads to a considerable reduction in sperm motility and alters sperm morfo-kinetic parameters, especially at a concentration of 5.6 mg/L. The results highlight a dose-dependent relationship between Gd exposure and the prevalence of specific malformation types in Mytilus embryos, further providing crucial insights into the potential risks imposed by this rare earth element.

Blue mussels are often abundant and widely distributed in polar marine coastal ecosystems. Because of their wide distribution, ecological importance, and relatively stationary lifestyle, bivalves have long been considered suitable indicators of ecosystem health and changes. Monitoring the population dynamics of blue mussels can provide information on the overall biodiversity, species interactions, and ecosystem functioning. In the present work, we combined the concept of liquid biopsy (LB), an emerging concept in medicine based on the sequencing of free circulating DNA, with the Oxford Nanopore Technologies (ONT) platform using a portable laboratory in a remote area. Our results demonstrate that this platform is ideally suited for sequencing hemolymphatic circulating cell-free DNA (ccfDNA) fragments found in blue mussels. The percentage of non-self ccfDNA accounted for >50 % of ccfDNA at certain sampling Sites, allowing the quick, on-site acquisition of a global view of the biodiversity of a coastal marine ecosystem. These ccfDNA fragments originated from viruses, bacteria, plants, arthropods, algae, and multiple Chordata. Aside from non-self ccfDNA, we found DNA fragments from all 14 blue mussel chromosomes, as well as those originating from the mitochondrial genomes. However, the distribution of nuclear and mitochondrial DNA was significantly different between Sites. Similarly, analyses between various sampling Sites showed that the biodiversity varied significantly within microhabitats. Our work shows that the ONT platform is well-suited for LB in sentinel blue mussels in remote and challenging conditions, enabling faster fieldwork for conservation strategies and resource management in diverse settings.

Pharmaceutical active compounds (PhACs) have raised concerns in the last decade due to their increased consumption and inadequate elimination during discharge, resulting in their introduction into water systems and potential significant threats to non-target organisms. However, few studies have investigated the sublethal impacts of PhAC exposure on marine invertebrates. Thus, the present study aimed to assess tissue-specific responses in Mytilus galloprovincialis to sodium lauryl sulfate (SLS), salicylic acid (SA), and caffeine (CAF) (4.0 mg/L, 4.0 mg/L and 2.0 μg/L, respectively). Short-term in vitro exposures with mussel digestive gland and gill tissues were conducted and biochemical responses related to antioxidant and detoxification capacity, cellular damage and neurotoxicity were assessed. The present results clearly showed significant differences in tissue sensitivity and biochemical responses to the contaminants tested. This study highlights the suitability of filter-feeder species as valuable model organisms for studying the sublethal effects of unintended environmental exposures to PhACs.

The present field-based study aimed to determine the levels of six potentially toxic metals (PTM)s (Cd, Cu, Fe, Ni, Pb, and Zn determined using a flame atomic-absorption spectrophotometer) using transplanted green-lipped mussel Perna viridis from a polluted site at Kampung Pasir Puteh (KPP) to unpolluted sites at Kampung Sungai Melayu (KSM) and Sungai Belungkor (SB) in the Johore Straits (SOJ), and to estimate the human health risks of the PTMs after the depuration periods. Interestingly, after 10 weeks of depuration in the two unpolluted sites, there were 55.6-88.4% and 51.3-91.7% reductions of the six PTMs after transplantation from KPP to SB and KSM, respectively. Lower risks of health assessments were recorded and judged on the present findings of significantly (p < 0.05) lower levels of safety guidelines, significantly (p < 0.05) lower values of target hazard quotient, and significantly (p < 0.05) lower values of estimated weekly intake, of all the six PTMs after 10 weeks of depuration of the transplanted polluted mussels to the two unpolluted sites in the SOJ. Thus, further reducing the noncarcinogenic risks of the PTMs to the consumers. From an aquacultural point of view, this depuration technique can be recommended to reduce the health risks of PTMs to mussel consumers.

Microcystins and nodularins, produced naturally by certain species of cyanobacteria, have been found to accumulate in aquatic foodstuffs such as fish and shellfish, resulting in a risk to the health of the seafood consumer. Monitoring of toxins in such organisms for risk management purposes requires the availability of certified matrix reference materials to aid method development, validation and routine quality assurance. This study consequently targeted the preparation of a mussel tissue reference material incurred with a range of microcystin analogues and nodularins. Nine targeted analogues were incorporated into the material as confirmed through liquid chromatography with tandem mass spectrometry (LC-MS/MS), with an additional 15 analogues detected using LC coupled to non-targeted high resolution mass spectrometry (LC-HRMS). Toxins in the reference material and additional source tissues were quantified using LC-MS/MS, two different enzyme-linked immunosorbent assay (ELISA) methods and with an oxidative-cleavage method quantifying 3-methoxy-2-methyl-4-phenylbutyric acid (MMPB). Correlations between the concentrations quantified using the different methods were variable, likely relating to differences in assay cross-reactivities and differences in the abilities of each method to detect bound toxins. A consensus concentration of total soluble toxins determined from the four independent test methods was 2425 ± 575 µg/kg wet weight. A mean 43 ± 9% of bound toxins were present in addition to the freely extractable soluble form (57 ± 9%). The reference material produced was homogenous and stable when stored in the freezer for six months without any post-production stabilization applied. Consequently, a cyanotoxin shellfish reference material has been produced which demonstrates the feasibility of developing certified seafood matrix reference materials for a large range of cyanotoxins and could provide a valuable future resource for cyanotoxin risk monitoring, management and mitigation.

Tetrodotoxins (TTXs), the pufferfish venom traditionally associated with Indo-Pacific area, has been reported during last decades in ever wider range of marine organisms and ever more geographical areas, including shellfish in Europe. Wild mussels (Mytilus galloprovincialis) grown in the Marche Region (N Adriatic Sea, Italy) were shown to be prone to TTX contamination during the warm season, with a suspected role of Vibrio alginolyticus characterized by non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS)-encoding genes. This work aimed to deepen the knowledge about the toxin\'s origin and the way through which it accumulates in mussels. A two-year study (spring-summer 2020-2021) confirmed the recurrent presence of TTX (11-68 µg kg-1) in the official monitored natural mussel beds of the Conero Riviera. During 2021, a supplementary nonroutine monitoring of a natural mussel bed in the same area was carried out weekly from June until August for TTXs and/or the presence of V. alginolyticus. Biotic (mussels, mesozooplankton, worms and phytoplankton); abiotic (water and sediment) matrices and phytoplankton assemblage characterizations were studied. Mussels showed relevant TTX contamination levels (9-296 µg kg-1) with extremely rapid TTX accumulation/depletion rates. The toxin presence in phytoplankton and its distribution in the different mussel tissues supports its possible exogenous origin. The V. alginolyticus count trend overlaps that of TTX contamination in mussels, and similar trends were reported also for some phytoplankton species. The role of V. alginolyticus carrying NRPS or PKS genes as a possible TTX source and of phytoplankton as a \"potential vector\" should therefore be further investigated.

The seasonal pattern of microplastics (MPs) contamination of the French littoral area of the Pertuis Charentais, one of the main French shellfish production regions, was assessed for the first time, between May 2019 and May 2020 at four different sites. The reference site was located at \"Ile de Ré\" and the other sites were located in the estuaries of the Sèvre Niortaise, Charente and Seudre rivers. Both blue mussels (Mytilus edulis) and Pacific oysters (Magallana gigas), that are considered sentinel species for the quality of the marine environment were analysed, along with sediment and seawater samples. MPs were extracted from each sample, counted, measured and sorted by colour and type. Micro-Raman spectroscopy was used to determine the proportion of confirmed MPs and the polymer types. The results showed that the contamination of mussels by fibres and fragments (1.9 ± 2.1 MPs/g ww) was significantly higher than for oysters (0.4 ± 0.4 MPs/g ww). Specifically, the contamination by fibres in both species was significantly greater than the contamination by fragments. Significant variations of MPs contamination were observed across the seasons and sites in bivalves, and depended on the species and the type of MPs (fibres or fragments). Mean concentrations of MPs measured in water and sediment were 0.007 MPs/L and 210 MPs/kg dw, respectively. Finally, blue was the dominant colour for fibres (79 %) and fragments (81 %). Blue fragments were mainly made of PS (70 %) followed by PC (18 %) and PP, PA or PLA (3 %) whereas blue fibres were mainly made of PA (80 %) followed by PET (13 %) or PP (7 %). This rare environmental case study of long-term chronic exposure of farming areas to MPs provides new knowledge on in situ variations of plastic fibres and fragments contamination throughout the seasons.

A quantification of total mercury (Hgtot) and methylmercury (MeHg) concentrations in sediment and mussels was carried out in the east basin of the Orbetello lagoon in order to assess their bioaccumulation potential. The sediment was sampled in four macroareas, mussels were transplanted in the same sites and collected after seven weeks. The results show that Hgtot concentrations in sediments exceeded (0.21-16.9 mg/kg dry weight (dw)) the environmental quality standard of the Italian legislation (0.3 mg/kg dw). The Hgtot concentration in mussels (0.050-0.324 mg/kg wet weight (ww)) does not exceed the limit values (0.5 mg/kg ww) of the European food legislation. The biota-sediment accumulation factors (BSAFs) derived for MeHg (80-306.7) and a biomagnification factor (BMF) greater than 1 for Hgtot demonstrate that in the lagoon, these compounds can be transferred in the upper levels of the trophic chain and pose a risk to human health.