[Departement_IRSTEA]Eaux [TR1_IRSTEA]BELCA | International audience | The separate and combined in vitro toxic effects of antibiotics (ciprofloxacin, erythromycin, novobiocin, oxytetracycline, sulfamethazole and trimethoprim) commonly found in urban wastewater effluents were assessed on the immune parameters of Elliptio complanata at environmentally relevant concentrations. The observed responses were then compared to those produced by the physicochemical-treated wastewater effluent of a major city before and after the removal of microorganisms. Most of the selected antibiotics, separately and as mixture, induced changes in immune responses. The removal of microorganisms and fine particles from the effluent increased or decreased the resulting immunotoxic effects, depending of the observed parameter. The immunotoxic effects of erythromycin, sulfamethoxazole and trimethoprim were closely associated to the antibiotic mixture and the filtered effluent. In conclusion, the data revealed that the removal of fine particles and microorganisms from municipal effluents can alter the toxic nature of the effluent that is closely associated with the cumulative effects of antibiotics.

Mussels (Mytilus galloprovincialis) were collected from the locality in the vinicity of the Institute for Marine Research in Kotor (Montenegro, Yugoslavia). The concentration of Cu (copper), Cr (chromium), Fe (iron), Zn (zinc), Ni (nickel), Mn (manganese), Pb (lead), Cd (cadmium), Al (aluminium), Ca (calcium) and Mg (magnesium) were determined in whole mulles, gills, pedal muscle and in rest of soft tissues. The data obtained in this research were compared with the results reported by other investigators.

Ocean acidification may increase the risk of disease outbreaks that would challenge the future persistence of marine organisms if their immune system and capacity to produce vital structures for survival (e.g., byssus threads produced by bivalves) are compromised by acidified seawater. These potential adverse effects may be exacerbated by microplastic pollution, which is forecast to co-occur with ocean acidification in the future. Thus, we evaluated the impact of ocean acidification and microplastics on the health of a mussel species (Mytilus coruscus) by assessing its physiological performance, immunity and byssus properties. We found that ocean acidification and microplastics not only reduced hemocyte concentration and viability due to elevated oxidative stress, but also undermined phagocytic activity of hemocytes due to lowered energy budget of mussels, which was in turn caused by the reduced feeding performance and energy assimilation. Byssus quality (strength and extensibility) and production were also reduced by ocean acidification and microplastics. To increase the chance of survival with these stressors, the mussels prioritized the synthesis of some byssus proteins (Mfp-4 and Mfp-5) to help maintain adhesion to substrata. Nevertheless, our findings suggest that co-occurrence of ocean acidification and microplastic pollution would increase the susceptibility of bivalves to infectious diseases and dislodgement risk, thereby threatening their survival and undermining their ecological contributions to the community.

Amyl salicylate (AS) is a fragrance massively used as a personal care product and following the discharged in wastewaters may end up in the aquatic environment representing a potential threat for the ecosystem and living organisms. AS was recently detected in water of the Venice Lagoon, a vulnerable area continuously subjected to the income of anthropogenic chemicals. The lagoon is a relevant area for mollusc farming, including the Mediterranean mussels (Mytilus galloprovincialis) having an important economic and ecological role. Despite high levels of AS occurred in water of the Lagoon of Venice, no studies investigated the possible consequences of AS exposures on species inhabiting this ecosystem to date. For the first time, we applied a multidisciplinary approach to investigate the potential effects of the fragrance AS on Mediterranean mussels. To reach such a goal, bioaccumulation, cellular, biochemical, and molecular analyses (RNA-seq and microbiota characterization) were measured in mussels treated for 7 and 14 days with different AS Venice lagoon environmental levels (0.1 and 0.5 μg L⁻¹). Despite chemical investigations suggested low AS bioaccumulation capability, cellular and molecular analyses highlighted the disruption of several key cellular processes after the prolonged exposures to the high AS concentration. Among them, potential immunotoxicity and changes in transcriptional regulation of pathways involved in energy metabolism, stress response, apoptosis and cell death regulations have been observed. Conversely, exposure to the low AS concentration demonstrated weak transcriptional changes and transient increased representation of opportunistic pathogens, as Arcobacter genus and Vibrio aestuarianus. Summarizing, this study provides the first overview on the effects of AS on one of the most widely farmed mollusk species.

Microplastic (MP) contamination is present in the entire marine environment from the sediment to the water surface and down to the deep sea. This ubiquitous presence of MP particles opens the possibility for their ingestion by nearly all species in the marine ecosystem. Reports have shown that MP particles are present in local commercial seafood species leading to the possible human ingestion of these particles. However, due to a lack of harmonized methods to identify microplastics (MPs), results from different studies and locations can hardly be compared. Hence, this study was aimed to detect, quantify, and estimate MP contamination in commercially important mussels originating from 12 different countries distributed worldwide. All mussels were obtained from supermarkets and were intended for human consumption. Using a combinatorial approach of focal plane array (FPA)-based micro- Fourier-transform infrared (FTIR) spectroscopy and micro-Raman spectroscopy allowed the detection and characterization of MP down to a size of 3 μm in the investigated mussels. Further, a gentle sample purification method based on enzymes has been modified in order to optimize the digestion of organic material in mussels. A random forest classification (RFC) approach, which allows a rapid discrimination between different polymer types and thus fast generation of data on MP abundance and size distributions with high accuracy, was implemented in the analytical pipeline for IR spectra. Additionally, for the first time we also applied a RFC approach for the automated characterization of Raman spectra of MPs.

Microplastics represent an emerging environmental issue and have been found almost everywhere including seafood, raising a great concern about the ecological and human health risks they pose. This study addressed the common technical challenges in the assessment of microplastics in seafood by developing an improved protocol based on Raman spectroscopy and using the green-lipped mussel Perna viridis and the Japanese jack mackerel Trachurus japonicus as the test models. Our findings identified a type of stainless-steel filter membranes with minimal Raman interference, and a combination of chemicals that achieved 99–100% digestion efficiency for both organic and inorganic biomass. This combined chemical treatment reached 90–100% recovery rates for seven types of microplastics, on which the surface modification was considered negligible and did not affect the accuracy of polymer identification based on Raman spectra, which showed 94–99% similarity to corresponding untreated microplastics. The developed extraction method for microplastics was further combined with an automated Raman mapping approach, from which our results confirmed the presence of microplastics in P. viridis and T. japonicus collected from Hong Kong waters. Identified microplastics included polypropylene, polyethylene, polystyrene and poly(ethylene terephthalate), mainly in the form of fragments and fibres. Our protocol is applicable to other biological samples, and provides an improved alternative to streamline the workflow of microplastic analysis for routine monitoring purposes.

Gymnodimine A has been found in mollusks obtained along the whole northern coast of Spain from April 2017 to December 2019. This is the first time that this toxin is detected in mollusks from the Atlantic coast of Europe. The prevalence of the toxin was, in general, low, being detected on average in approximately 6% of the obtained samples (122 out of 1900). The concentrations recorded were also, in general, low, with a median of 1.3 μg kg⁻¹, and a maximum value of 23.93 μg kg⁻¹. The maxima of prevalence and concentration were not geographically coincident, taking place the first at the easternmost part of the sampled area and the second at the westernmost part. In most cases (>94%), gymnodimine A and 13-desmethyl spirolide C were concurrently detected, suggesting that Alexandrium ostenfeldii could be the responsible producer species. The existence of cases in which gymnodimine A was detected alone suggests also that a Karenia species could also be involved. The geographical heterogeneity of the distribution suggests that blooms of the producer species are mostly local. Not all bivalves are equally affected, clams being less affected than mussels, oysters, and razor clams. Due to their relatively low toxicity, and their low prevalence and concentration, it seems that these toxins do not pose an important risk for the mollusk consumers in the area.

The impacts of microplastic particulates in benthic freshwater organisms have been largely unexplored despite abundant plastic accumulation in the sediments of these systems. We investigated the uptake of plastic particles by benthic filter feeding quagga mussels (Dreissena bugensis) and associated toxicity exhibited through impacts on mortality, filtration rate, reproduction and oxygen consumption. Matrix Assisted Laser Desorption/Ionization Imaging Mass Spectrometry (MALDI-IMS) technology was used to assess the microplastic inclusion. For this purpose, quagga mussels were exposed to four treatments ranging from 0.0 to 0.8 g/L of a high density fluorescent red polyethylene powder in the size range of 10–45 μm for 24-h, and the targeted endpoints were quantified. Identification of several micrograms of microplastics in the digestive tract suggests rapid clearance from the water column by filtering. At the higher concentrations, about 95% of the microplastics ingested remained in the mussels after 24-h. Microplastics were found in the gills which correlated with decreasing filtration rate at higher microplastic concentrations. Despite large-scale ingestion, plastic exposure did not affect survivorship, reproduction rates, or oxygen consumption in the period examined. MALDI-IMS identified unique mass spectra that correlated with microplastic inclusion. This research suggests that microplastics can impair feeding through decreased filtration rates of filter feeding organisms, potentially resulting in a reduction of overall fitness over time and that MALDI-IMS may have the potential to identify microplastics and changes in tissue at the borders of plastic inclusion.

The exposure risk of metal-based nanoparticles (NPs) to marine organisms and related food safety have attracted increasing attention, but the actual concentrations of these NPs in seawater and marine organisms are unknown. In this work, single particle inductively coupled plasma-mass spectrometry (spICP-MS) was used to quantify the concentrations and size distributions of NPs in different marine mollusks (oysters, mussels, scallops, clams, and ark shells) from an offshore aquaculture farm. Results showed that Ti, Cu, Zn, and Ag bearing NPs were detected in all the five mollusks with the mean sizes at 65.4–70.9, 72.2–89.6, 97.8–108.3, and 42.9–51.0 nm, respectively. The particle concentrations of Ti, Cu, Zn, and Ag bearing NPs in all mollusks (0.88–3.26 × 10⁷ particles/g fresh weight) were much higher than that in the seawater (0.46–0.79 × 10⁷ particles/mL), suggesting bio-accumulation of NPs. For all the five mollusks, Ag bearing NPs had the highest number-based bioconcentration factors (NBCFs) in all the tested NPs due to the smallest mean size of Ag bearing NPs in seawater (30.5 nm). In addition, the clams exhibited the lowest NBCFs of the four NPs than other mollusks. All four NPs were mainly accumulated in the gill and digestive gland, and could transfer to adductor muscle of all mollusks. Although all the four metals (Ti, Cu, Zn, Ag) in mollusks were safe for human consumption by the estimated daily intake (EDI) analysis, the risk of NPs remaining in the mollusks should be further considered when evaluating the toxicity of metals for human health. The findings could improve our understanding on the distribution and health risk of NPs in marine mollusks under offshore aquaculture.

The majority of plastics present in the marine environment are microplastics (MPs, <5 mm). Suspension filter feeders are susceptible species to MPs ingestion. Once ingested MPs can be eliminated packed in fecal pellets, or they can be accumulated within tissues, and likely be transferred along the food web. The research on MPs is hampered by the difficulty on their quantification and the lack of standardized methodologies. Indeed, limited information exits about the capacity of marine organisms to ingest, accumulate and eliminate MPs. In this work we investigated the uptake, elimination and accumulation of MPs (irregularly shaped particles of high density polyethylene, ≤22 μm) in mussel. Mussels were exposed to two concentrations of MPs (2 and 4 mm3 l−1), and their uptake, elimination and accumulation in digestive gland was investigated. The results showed that the uptake of MPs increased at the high concentration tested, and that mussels cleared MPs at the same extent than a food item (microalgae) of similar size. Small MPs (2–4 μm) were less efficiently cleared than the larger ones. Large MPs (>10 μm) were faster eliminated than the smaller ones. The global balance showed that after 6 days of depuration mussels eliminated ≈85% of the MPs cleared, and that ≈2–6% of the MPs cleared remained in the digestive gland, essentially those <6 μm. We recorded a long retention time for MPs, contrasting with the lower times assumed to be necessary to empty mussel's gut before quantifying MPs. Our study emphasized the gap of knowledge on the feeding behaviour of mussels in relation to MPs, and the necessity to investigate it in different marine species, and under different exposure scenarios.