Microplastic ingestion poses a significant concern for a plethora of marine organisms due to its widespread presence in marine ecosystems. Despite growing scientific interest, the effects on marine biota are not yet well understood. This study investigates the ingestion of microplastics (MPs) by mussels from various marine environments and assesses the associated effects that can be induced by MPs and associated toxic chemicals. Biomarkers of oxidative stress (catalase, lipid peroxidation), biotransformation (glutathione S-transferase), genotoxicity (micronuclei frequency) and neurotoxicity (acetylcholinesterase) were employed. Mussels, considered reliable bioindicators of MPs pollution, were sampled by hand from diverse locations under varied anthropogenic pressures, including a highly touristic Marine Protected Area (MPA) in the Ionian Sea, a mussel farm and a fish farm in the Aegean Sea. The results revealed the highest MP ingestion in mussels from the fish farm [0.21 ± 0.04 (SE) MPs/g or 0.63 ± 0.12 (SE) MPs/Ind.], likely due to plastic aquaculture equipment use. Stereoscopic observation revealed fibers, as the predominant shape of ingested MPs across all sites, and μFTIR polymer identification revealed the presence of various types, with polyethylene (PE) and polyamide (PA) being the most abundant. Significant physiological alterations in mussels related to MP ingestion levels were observed through biomarkers indicative of oxidative stress and biotransformation, as well as the Integrated Biomarker Response (IBR index). However, laboratory experiments with mussels exposed to controlled increasing PE concentrations for four weeks, did not show significant effects triggered by the PE ingestion, possibly indicating other environmental factors, such as contaminants from aquaculture environments, may influence biomarker levels in the field. Despite the observed effects, MP ingestion rates in mussels from the field were relatively low compared to other studies. Future research should continue to investigate the interactions between MPs and marine organisms in diverse environments to better understand and mitigate their impacts.

The effect of water acidification in combination with normoxia or hypoxia on the antioxidant capacity and oxidative stress markers in gills and hemolymph of the Mediterranean mussel (Mytilus galloprovincialis), as well as on gill microstructure, has been evaluated through an in vivo experiment. Mussels were exposed to a low pH (7.3) under normal dissolved oxygen (DO) conditions (8 mg/L), and hypoxia (2 mg/L) for 8 days, and samples were collected on days 1, 3, 6, and 8 to evaluate dynamic changes of physiological responses. Cytoplasmic concentrations of reactive oxygen species (ROS) and levels of DNA damage were measured in hemocytes, while the activity of catalase (CAT) and superoxide dismutase (SOD) and histopathological changes were assessed in gills. The results revealed that while water acidification did not significantly affect the activity of SOD and CAT in gills under normoxic and hypoxic conditions, there was a trend towards suppression of CAT activity at the end of the experimental period (day 8). Similarly, we did not observe increased formation of ROS in hemocytes or changes in the levels of DNA damage during the experimental period. These results strongly suggest that the oxidative stress response system in mussels is relatively stable to experimental conditions of acidification and hypoxia. Experimental acidification under normoxia and hypoxia caused changes to the structure of the gills, leading to various histopathological alterations, including dilation, hemocyte infiltration into the hemal sinuses, intercellular edema, vacuolization of epithelial cells in gill filaments, lipofuscin accumulation, changes in the shape and adjacent gill filaments, hyperplasia, exfoliation of the epithelial layer, necrosis, swelling, and destruction of chitinous layers (chitinous rods). Most of these alterations were reversible, non-specific changes that represent a general inflammatory response and changes in the morphology of the gill filaments. The dynamics of histopathological alterations suggests an active adaptive response of gills to environmental stresses. Taken together, our data indicate that Mediterranean mussels have a relative tolerance to water acidification and hypoxia at tissue and cellular levels.

The overproduction and mismanagement of plastics has led to the accumulation of these materials in the environment, particularly in the marine ecosystem. Once in the environment, plastics break down and can acquire microscopic or even nanoscopic sizes. Given their sizes, microplastics (MPs) and nanoplastics (NPs) are hard to detect and remove from the aquatic environment, eventually interacting with marine organisms. This research mainly aimed to achieve the aggregation of micro- and nanoplastics (MNPs) to ease their removal from the marine environment. To this end, the size and stability of polystyrene (PS) MNPs were measured in synthetic seawater with the different components of the technology (ionic liquid and chitosan). The MPs were purchased in their plain form, while the NPs displayed amines on their surface (PS NP-NH2). The results showed that this technology promoted a significant aggregation of the PS NP-NH2, whereas, for the PS MPs, no conclusive results were found, indicating that the surface charge plays an essential role in the MNP aggregation process. Moreover, to investigate the toxicological potential of MNPs, a mussel species (M. galloprovincialis) was exposed to different concentrations of MPs and NPs, separately, with and without the technology. In this context, mussels were sampled after 7, 14, and 21 days of exposure, and the gills and digestive glands were collected for analysis of oxidative stress biomarkers and histological observations. In general, the results indicate that MNPs trigger the production of reactive oxygen species (ROS) in mussels and induce oxidative stress, making gills the most affected organ. Yet, when the technology was applied in moderate concentrations, NPs showed adverse effects in mussels. The histological analysis showed no evidence of MNPs in the gill\'s tissues.

Tracking PFAS in ecosystems is challenging. In this context, monitoring programs are crucial to fill data gaps, especially in marine environments, which are the ultimate outlets for these forever chemicals. The 2021 chemical contamination monitoring campaign along the French Mediterranean coast established a baseline for PFAS concentrations in mussels, with 90 % of measurements below quantification limits. When detected, long-chain PFCA\'s were predominant. Spatial distribution patterns suggested continuous PFAS inputs and complex dynamics, shaped by the influence of large watersheds and rivers (Rhône, Aude, Huveaune). Lapeyrade shallow lagoon stood out as the most contaminated site. Similar PFAS profiles in connected sites implied shared sources but raised questions about accumulation processes in mussels. While certain sites had evident sources (e.g., military airbase for Palo lagoon), others remained uncertain (e.g., Toulon bay). Coastal stations (Banyuls, Cap Agde, Brégançon, Pampelonne) showed PFAS contamination without clear onshore sources, possibly due to insufficient transportation process understanding.

This study delves into the intricate interplay between ocean acidification (OA), metal bioaccumulation, and cellular responses using mussels (Mytilus galloprovincialis) as bioindicators. For this purpose, environmentally realistic concentrations of isotopically labelled metals (Cd, Cu, Ag, Ce) were added to investigate whether the OA increase would modify metal bioaccumulation and induce adverse effects at the cellular level. The study reveals that while certain elements like Cd and Ag might remain unaffected by OA, the bioavailability of Cu and Ce could potentially escalate, leading to amplified accumulation in marine organisms. The present findings highlight a significant rise in Ce concentrations within different mussel organs under elevated pCO2 conditions, accompanied by an increased isotopic fractionation of Ce (140/142Ce), suggesting a heightened potential for metal accumulation under OA. The results suggested that OA influenced metal accumulation in the gills of mussels. Conversely, metal accumulation in the digestive gland was unaffected by OA. The exposure to both trace metals and OA affects the biochemical responses of M. galloprovincialis, leading to increased metabolic capacity, changes in energy reserves, and alterations in oxidative stress markers, but the specific effects on other biomarkers (e.g., lipid peroxidation, some enzymatic responses or acetylcholinesterase activity) were not uniform, suggesting complex interactions between the stressors and the biochemical pathways in the mussels.

In mussel hatchery systems, the settlement process is a crucial element influencing seed yield. The current study assayed the influence of five densities of competent pediveliger larvae on settlement success and post-larvae production. We showed an inverse relationship between density and settlement efficiency, e.g., an attachment success of 99.4% at the lowest density (35 larvae/cm2) but only 9% at the highest density (210 larvae/cm2). However, post-larvae production was higher at intermediate larvae densities (70 larvae/cm2). The reimplementation of treatments upon post-larvae density after 6 weeks post settlement showed that the lowest-density groups bore both the highest post-larvae growth rate (22.24 ± 4.60 µm/day) and the largest head batch (48% of the size distribution), as compared to the higher-post-larvae-density groups. These results highlight the importance of optimizing both pediveliger larvae density and post-larvae density, to maximize high-quality seed yield in local hatcheries. Current rearing technologies would assure a timely commercial seed production to protect natural sea rocky beds in Alboran Sea coasts.

In recent years, the presence of pharmaceuticals and microplastics (MPs) in aquatic ecosystems has raised concerns about their environmental impact. This study explores the combined effects of caffeine, a common pharmaceutical pollutant, and MPs on the marine mussel Mytilus galloprovincialis. Caffeine, at concentrations of 20.0 μg L-1, and MPs (1 mg L-1, 35-50 μm size range), was used to mimic real-world exposure scenarios. Two hundred M. galloprovincialis specimens were divided into four groups: caffeine, MPs, Mix (caffeine + MPs), and Control. After a two-week acclimation period, the mollusks were subjected to these pollutants in oxygen-aerated aquariums under controlled conditions for 14 days. Histopathological assessments were performed to evaluate gill morphology. Cellular volume regulation and digestive gland cell viability were also analyzed. Exposure to caffeine and MPs induced significant morphological changes in M. galloprovincialis gills, including cilia loss, ciliary disk damage, and cellular alterations. The chitinous rod supporting filaments also suffered damage, potentially due to MP interactions, leading to hemocyte infiltration and filament integrity compromise. Hemocytic aggregation suggested an inflammatory response to caffeine. In addition, viability assessments of digestive gland cells revealed potential damage to cell membranes and function, with impaired cell volume regulation, particularly in the Mix group, raising concerns about nutrient metabolism disruption and organ function compromise. These findings underscore the vulnerability of M. galloprovincialis to environmental pollutants and emphasize the need for monitoring and mitigation efforts. RESEARCH HIGHLIGHTS: The synergy of caffeine and microplastics (MPs) in aquatic ecosystems warrants investigation. MPs and caffeine could affect gill morphology of Mytilus galloprovincialis. Caffeine-exposed cells had lower viability than the control group in the NR retention test. MPs and mix-exposed cells struggled to recover their volume.

Microplastic contamination is a growing marine environmental issue with possible consequences for seafood safety. Filter feeders are the target species for microplastic (MPs) pollution because they filter large quantities of seawater to feed. In the present study, an experimental contamination of Mytilus galloprovincialis was conducted using a mixture of the main types of MPs usually present in the seawater column (53% filaments, 30% fragments, 3% granules) in order to test the purification process as a potential method for removing these contaminants from bivalves intended for human consumption. A set of molecular biomarkers was also evaluated in order to detect any variations in the expression levels of some genes associated with biotransformation and detoxification, DNA repair, cellular response, and the immune system. Our results demonstrate that: (a) the purification process can significantly reduce MP contamination in M. galloprovincialis; (b) a differential expression level has been observed between mussels tested and in particular most of the differences were found in the gills, thus defining it as the target organ for the use of these biomarkers. Therefore, this study further suggests the potential use of molecular biomarkers as an innovative method, encouraging their use in next-generation marine monitoring programs.

The presence of doubly uniparental inheritance (DUI) in bivalves represents a unique mode of mitochondrial transmission, whereby paternal (male-transmitted M-type) and maternal (female-transmitted F-type) haplotypes are transmitted to offspring separately. Male embryos retain both haplotypes, but the M-type is selectively removed from females. Due to the presence of heteroplasmy in males, mtDNA can recombine resulting in a \'masculinized\' haplotype referred to as Mf-type. While mtDNA recombination is usually rare, it has been recorded in multiple mussel species across the Northern Hemisphere. Given that mitochondria are the powerhouse of the cell, different mtDNA haplotypes may have different selective advantages under diverse environmental conditions. This may be particularly important for sperm fitness and fertilization success. In this study we aimed to i) determine the presence, prevalence of the Mf-type in Australian blue mussels (Mytilus sp.) and ii) investigate the effect of Mf-mtDNA on sperm performance (a fitness correlate). We found a high prevalence of recombined mtDNA (≈35 %) located within the control region of the mitochondrial genome, which occurred only in specimens that contained Southern Hemisphere mtDNA. The presence of two female mitotypes were identified in the studied mussels, one likely originating from the Northern Hemisphere, and the other either representing the endemic M. planulatus species or introduced genotypes from the Southern Hemisphere. Despite having recombination events present in a third of the studied population, analysis of sperm performance indicated no difference in fertilization success related to mitotype.

In recent years, metabolomics has become a valuable new resource in environmental monitoring programs based on the use of bio-indicators such as Mytilus galloprovincialis. The reproductive system is extremely susceptible to the effects of environmental pollutants, and in a previous paper, we showed metabolomic alterations in mussel spermatozoa exposed to metal chlorides of copper, nickel, and cadmium, and the mixture with these metals. In order to obtain a better overview, in the present work, we evaluated the metabolic changes in the male gonad under the same experimental conditions used in the previous work, using a metabolomic approach based on GC-MS analysis. A total of 248 endogenous metabolites were identified in the male gonads of mussels. Statistical analyses of the data, including partial least squares discriminant analysis, enabled the identification of key metabolites through the use of variable importance in projection scores. Furthermore, a metabolite enrichment analysis revealed complex and significant interactions within different metabolic pathways and between different metabolites. Particularly significant were the results on pyruvate metabolism, glycolysis, and gluconeogenesis, and glyoxylate and dicarboxylate metabolism, which highlighted the complex and interconnected nature of these biochemical processes in mussel gonads. Overall, these results add new information to the understanding of how certain pollutants may affect specific physiological functions of mussel gonads.