Geographical and ecological factors affect microplastic body burden in marine fish at global scale.

Microplastic (MP) contamination has been identified as a worrisome environmental issue at the global level. Fish are the taxonomic group more extensively investigated to assess MP contamination in marine environment. A large variability in MP bioaccumulation (i.e., body burden) was reported in fish but to date there is a dearth of information concerning the drivers underlying this process. The present systematic review aimed at summarizing the results of the scientific literature on MP body burden in the digestive tract of marine fish to quantitatively shed light on the contribution of different geographical (i.e., latitudinal origin of the sample, distance from the coastline and field- or marked-collected) and ecological (i.e., trophic strategy, milieu, and body size) factors driving bioaccumulation. The mean (±SE) MPs/individual was 4.13 ± 2.87, and the mean MPs/ww (i.e., MPs/g) was 5.92 ± 0.94. Overall, MP abundance expressed as MPs/individual of fish from tropical areas was significantly higher compared to the other latitudinal bands, with species sampled close to the coastline that accumulated a larger number of MPs compared to those collected offshore. Neither the trophic strategy, nor the milieu and the market or field origin of fish explained the MP body burden. However, fish body size resulted as a determinant of MP body burden (as MPs/individual), with small fish accumulating a lower amount of MPs compared to larger ones. Qualitatively, but not statistically significant, similar results were generally obtained for MPs/ww, except for an opposite, and significant, variation according to species body size. Our findings showed that geographical, rather than ecological factors represent the main drivers of MP body burden in marine fish, suggesting that environmental variables and/or local pollution sources mainly contribute to explaining the large variability underlying the ingestion and bioaccumulation processes of these contaminants.

Potential Effects of Environmental and Occupational Exposure to Microplastics: An Overview of Air Contamination.

Microplastics (MPs) are now ubiquitous environmental contaminants that lead to unavoidable human exposure; they have received increasing attention in recent years and have become an emerging area of research. The greatest concern is the negative impacts of MPs on marine, fresh-water, and terrestrial ecosystems, as well as human health, to the extent that the World Health Organization (WHO) calls for increased research and standardized methods to assess exposure to MPs. Many countries and international organizations are implementing or proposing legislation in this regard. This review aims to summarize the current state of legislation, indoor and outdoor contamination, and potential human health risk due to exposure to airborne MPs, considering that occupational exposure to MPs is also becoming a growing area of concern. Even though research regarding MPs has continuously increased in the last twenty years, the effects of MPs on human health have been scarcely investigated, and toxicity studies are still limited and not directly comparable, due to the lack of standardized studies in this field.

Indigo-dyed cellulose fibers and synthetic polymers in surface-feeding seabird chick regurgitates from the Gulf of Alaska.

We provide evidence of anthropogenic materials ingestion in seabirds from a remote oceanic area, using regurgitates obtained from black-legged kittiwake (Rissa tridactyla) chicks from Middleton Island (Gulf of Alaska, USA). By means of GPS tracking of breeding adults, we identified foraging grounds where anthropogenic materials were most likely ingested. They were mainly located within the continental shelf of the Gulf of Alaska and near the Alaskan coastline. Anthropogenic cellulose fibers showed a high prevalence (85 % occurrence), whereas synthetic polymers (in the micro- and mesoplastics dimensional range) were less frequent (20 %). Most fibers (60 %) were blue and we confirmed the presence of indigo-dyed cellulosic fibers, characteristic of denim fabrics. In terms of mass, contamination levels were 0.077 µg g-1 wet weight and 0.009 µg g-1 wet weight for anthropogenic microfibers and synthetic polymers, respectively. These results represent the only recent report of contamination by anthropogenic fibers in seabirds from the Gulf of Alaska.

Altitudinal variation of microplastic abundance in lakeshore sediments from Italian lakes.

Microplastic (MP) contamination represents an issue of global concern for both aquatic and terrestrial ecosystems, but only in recent years, the study of MPs has been focused on freshwaters. Several monitoring surveys have detected the presence of a wide array of MPs differing in size, shape, and polymer composition in rivers and lakes worldwide. Because of their role of sink for plastic particles, the abundance of MPs was investigated in waters, and deep and shoreline sediments from diverse lakes, confirming the ubiquity of this contamination. Although diverse factors, including those concerning anthropogenic activities and physical characteristics of lakes, have been supposed to affect MP abundances, very few studies have directly addressed these links. Thus, the aim of the present study was to explore the levels of MP contamination in mountain and subalpine lakes from Northern Italy. Fourteen lakes dislocated at different altitudes and characterized by dissimilar anthropic pressures were visited. Lakeshore sediments were collected close to the drift line to assess MPs contamination. Our results showed the presence of MPs in lakeshore sediments from all the lakes, with a mean (± standard deviation) expressed as MPs/Kg dry sediment accounting to 14.42 ± 13.31 (range 1.57-61.53), while expressed as MPs/m2, it was 176.07 ± 172.83 (range 25.00-666.67). The MP abundance measured for Garda Lake was significantly higher compared to all the other ones (F1,13 = 7.344; P < 0.001). The pattern of contamination was dominated by fibers in all the lakes, but they were the main contributors in mountain lakes. These findings showed that the MP abundance varied according to the altitude of the lakes, with higher levels measured in subalpine lakes located at low altitudes and surrounded by populated areas.

Multi-level toxicity assessment of polylactic acid (PLA) microplastics on the cladoceran Daphnia magna.

The accumulation of plastics waste in the environment has raised a worrisome concern, moving the society to seek out for sustainable solutions, such as the transition from the use of fossil-based, conventional plastics to bioplastics (BPs). However, once in the environment bioplastics have the same probability to accumulate and experience weathering processes than conventional plastics, leading to the formation of microplastics (MPs). However, to date the information on the potential toxicity of MPs originated from the weathering of bioplastics is limited. Thus, this study aimed at investigating the adverse effects induced by the exposure to MPs made of a bioplastic polymer, the polylactic acid (PLA), towards the freshwater cladoceran Daphnia magna. Organisms were exposed for 21 days to three concentrations (0.125 µg/mL, 1.25 µg/mL and 12.5 µg/mL) of PLA microplastics (hereafter PLA-MPs). A multi-level approach was performed to investigate the potential effects through the biological hierarchy, starting from the sub-individual up to the individual level. At the sub-individual level, changes in the oxidative status (i.e., the amount of reactive oxygen species and the activity of antioxidant and detoxifying enzymes) and oxidative damage (i.e., lipid peroxidation) were explored. Moreover, the total caloric content as well as the content of protein, carbohydrate and lipid content assess were used to investigate the effects on energy reserves. At individual level the changes in swimming activity (i.e., distance moved and swimming speed) were assessed. Our results showed that the exposure to PLA-MPs induced a slight modulation in the oxidative status and energy reserves, leading to an increase in swimming behavior of treated individuals compared to control conspecifics. These results suggest that the exposure to MPs made of a bioplastic polymer can induce adverse effects similar to those caused by conventional polymers.

Exposure to Microplastics Made of Plasmix-Based Materials at Low Amounts Did Not Induce Adverse Effects on the Earthworm Eisenia foetida.

The implementation of recycling techniques represents a potential solution to the plastic pollution issue. To date, only a limited number of plastic polymers can be efficiently recycled. In the Italian plastic waste stream, the residual, non-homogeneous fraction is called 'Plasmix' and is intended for low-value uses. However, Plasmix can be used to create new materials through mechanical recycling, which need to be tested for their eco-safety. This study aimed to investigate the potential toxicity of two amounts (0.1% and 1% MPs in soil weight) of microplastics (MPs) made of naïve and additivated Plasmix-based materials (Px and APx, respectively) on the earthworm Eisenia foetida. Changes in oxidative status and oxidative damage, survival, gross growth rate and reproductive output were considered as endpoints. Although earthworms ingested both MP types, earthworms did not suffer an oxidative stress condition or growth and reproductive impairments. The results suggested that exposure to low amounts of both MPs can be considered as safe for earthworms. However, further studies testing a higher amount or longer exposure time on different model species are necessary to complete the environmental risk assessment of these new materials.

Comparison of the potential toxicity induced by microplastics made of polyethylene terephthalate (PET) and polylactic acid (PLA) on the earthworm Eiseniafoetida.

A growing number of studies have demonstrated that microplastic (MP) contamination is widespread in terrestrial ecosystems. A wide array of MPs made of conventional, fossil-based polymers differing in size and shape has been detected in soils worldwide. Recently, also MPs made of bioplastics have been found in soils, but there is a dearth of information concerning their toxicity on soil organisms. This study aimed at exploring the potential toxicity induced by the exposure for 28 days to irregular shaped and differently sized MPs made of a fossil-based (polyethylene terephthalate - PET) and a bioplastic (polylactic acid - PLA) polymer on the earthworm Eisenia foetida. Two amounts (1 g and 10 g/kg of soil, corresponding to 0.1% and 1% of soil weight) of both MP types were administered to the earthworms. A multi-level approach was used to investigate the MP-induced effects at sub-individual and individual level. Changes in the activity of antioxidant and detoxifying enzymes, as well as in lipid peroxidation levels, were investigated at specific time-points (i.e., 7, 14, 21 and 28 days) as sub-individual responses. Histological analyses were performed to assess effects at tissue level, while the change in digging activity was considered as a proxy of behavioral effects. Earthworms ingested MPs made of both the polymers. MPs made of PET did not induce any adverse effect at none of the biological levels. In contrast, MPs made of PLA caused the modulation of earthworms' oxidative status as showed by a bell-shaped activity of superoxide dismutase coupled with an increase in glutathione peroxidase activity. However, neither oxidative and tissue damage, nor behavioral alteration occurred. These findings suggest that the exposure to bio-based MPs can cause higher toxicity compared to fossil-based MPs.

Exposure to polystyrene nanoplastics induced physiological and behavioral effects on the brittle star Ophiactis virens.

Nanoplastic contamination has become an issue of environmental concern but the information on the potential adverse effects of nanoplastics on marine ecosystems is still limited. Therefore, the aim of this work was to investigate the effects of the exposure to polystyrene nanoplastics (PS-NPs; 0.05, 0.5 and 5 µg/mL) on the brittles star Ophiactis virens. Diverse endpoints at different levels of biological organization were considered, including behavior, arm regeneration capacity and oxidative stress. PS-NPs were observed on the brittle star body surface but not in inner tissues. Accumulation of PS-NPs was observed in the pre-buccal cavity of animals exposed to 5 µg/mL PS-NPs which also displayed delayed righting activity and an oxidative stress condition. Nevertheless, no effect was observed on arm regeneration efficiency at any tested PS-NPs concentration. Overall, our results highlighted that prolonged exposure to high amounts of PS-NPs could interfere at least partially with the physiology of O. virens.

Interspecific competitive interactions affect body size and oxidative status of two nonnative salmonid species.

In fish, interspecific interactions between nonnative and other sympatric species are considered determinants in shaping species assemblages. Such interactions can also arise between nonnative fish species only, including salmonids such as the brown trout (Salmo trutta, Linnaeus, 1758) and the rainbow trout (Oncorhynchus mykiss, Walbaum, 1792), returning contrasting outcomes. The present manipulative experiment was aimed at exploring the effect of interspecific competition on the body growth and the oxidative status of parr (2 + -year-old individuals) of the brown trout and the rainbow trout. Allopatric (intraspecific competition) and sympatric (interspecific competition) populations of these species were experimentally recreated in two wild streams. At the end of a 2-month-long experiment, changes in specific growth rate (SGR), oxidative status (i.e., levels of reactive oxygen species and activity of antioxidant enzymes such as superoxide dismutase - SOD, catalase - CAT and glutathione peroxidase - GPx) and oxidative damage (i.e., lipid peroxidation) were investigated in brown and rainbow trout individuals maintained in allopatric or sympatric populations. Sympatric interactions between rainbow and brown trout parr resulted in a significant decrease in SGR of brown trout individuals only. Moreover, an overall modulation of the oxidative status, in terms of an increase in ROS levels coupled with the activation of SOD and CAT activity, occurred in brown trout individuals under sympatric conditions. These findings might suggest that, under sympatric conditions, parr of the rainbow trout are more competitive than brown trout for food acquisition. However, this competition affected the antioxidant defenses of the brown trout only, probably because of reduced ingestion of dietary antioxidants or increased physical activity and aggressive behavior. Thus, interspecific interactions can induce physiological and phenotypic effects on parr of nonnative salmonids, with potential consequences on the establishment of populations of these species in freshwater ecosystems.

Exposure to 3,4-methylenedioxymethamphetamine (MDMA) induced biochemical but not behavioral effects in Daphnia magna.

Among amphetamine like stimulants (ATS), the 3,4-methylenedioxymethamphetamine (MDMA) is often detected in sewage and surface waters, representing a potential threat for organisms because of its peculiar mechanism of action (i.e., stimulatory and hallucinogenic). The present study aimed at investigating biochemical (i.e., oxidative stress and energetic biomarkers) and behavioral (i.e., swimming activity) effects induced by a 21-days exposure to two concentrations (50 ng/L and 500 ng/L) of MDMA towards Daphnia magna. The amount of reactive oxygen species (ROS), the activity of antioxidant (SOD, CAT, GPx) and detoxifying (GST) enzymes and lipid peroxidation were measured as oxidative stress-related endpoints. Total energy content was estimated from the measurement of protein, carbohydrate and lipid content to assess energy reserves. The modulation of swimming activity was assessed as behavioral endpoint. Slight effects of MDMA exposure on oxidative stress responses and energy reserves were observed, while no alterations of the swimming behavior was noted.

Microplastics Contamination in Nonalcoholic Beverages from the Italian Market.

A growing number of studies have confirmed that microplastics (MPs) contamination represents a worrisome issue of global concern. MPs have been detected in the atmosphere, in aquatic and terrestrial ecosystems, as well as in the biota. Moreover, MPs have been recently detected in some food products and in drinking water. However, only limited information is currently available for beverages, although they are largely consumed by humans and might contribute to the ingestion of MPs. Thus, estimating the contamination in beverages represents a crucial step in assessing human MP ingestion. The aim of the present study was to explore the presence of MPs in nonalcoholic beverages, namely soft drinks and cold tea, of different brands purchased in supermarkets and to estimate the contribution of beverage consumption to MP ingestion by humans. The results of the present study confirmed the presence of MPs, mainly fibers, in most of the analyzed beverages, with a mean (± SEM) number of 9.19 ± 1.84 MPs/L. In detail, the number of MPs detected in soft drinks and cold tea was 9.94 ± 0.33 MPs/L and 7.11 ± 2.62 MPs/L, respectively. Our findings confirmed that beverage consumption can be considered one of the main pathways for MP ingestion by humans.

Impact of anthropogenic contamination on glacier surface biota.

Glaciers are ecosystems and they host active biological communities. Despite their remoteness, glaciers act as cold condensers where high precipitation rates and cold temperatures favor the deposition of pollutants. These contaminants include a broad range of substances, including legacy pollutants, but also compounds still largely used. Some of these compounds are monitored in the environment and their effects on the ecosystems are known, in contrast others can be defined as emerging pollutants since their presence and their impact on the environment are still poorly understood (e.g. microplastics, radionuclides). This review aim to provide an overview of the studies that have investigated the effects of pollutants on the supraglacial ecosystem so far. Despite the distribution of the pollutants in glacier environments has been discussed in several studies, no review paper has summarized the current knowledge on the effects of these substances on the ecological communities living in glacier ecosystems.

Polystyrene microplastics exposure modulated the content and the profile of fatty acids in the Cladoceran Daphnia magna.

A growing number of studies has shown that the exposure to microplastics (MPs) of different polymeric compositions can induce diverse adverse effects towards several aquatic species. The vast majority of such studies has been focused on the effects induced by the administration of MPs made by polystyrene (PS; hereafter PS-MPs). However, despite the increase in the knowledge on the potential toxicity of PS-MPs, there is a dearth of information concerning their role in affecting energy resources and/or their allocation. The present study aimed at exploring the impact of 21-days exposure to three concentrations (0.125, 1.25 and 12.5 µg mL-1) of PS-MPs of different sizes (1 and 10 µm) on fatty acids (FAs) profile of the freshwater Cladoceran Daphnia magna. The exposure to the highest tested concentration of PS-MPs induced an overall decrease in D. magna total FAs content, independently of the particle size. Moreover, a change in the accumulation of essential FAs by the diet was noted, with an enhanced synthesis of monounsaturated FAs-rich storage lipids. However, a sort of adaptation to counteract the adverse effects and to re-establish the FAs homeostasis was observed in individuals treated with high PS-MPs concentration, independently of their size. These results indicate that the exposure to PS-MPs could alter the allocation or induce changes in FAs composition in D. magna, with potential long-term consequences on life-history traits of this zooplanktonic species.

Fe3O4-Halloysite Nanotube Composites as Sustainable Adsorbents: Efficiency in Ofloxacin Removal from Polluted Waters and Ecotoxicity.

The present work aimed at decorating halloysite nanotubes (HNT) with magnetic Fe3O4 nanoparticles through different synthetic routes (co-precipitation, hydrothermal, and sol-gel) to test the efficiency of three magnetic composites (HNT/Fe3O4) to remove the antibiotic ofloxacin (OFL) from waters. The chemical-physical features of the obtained materials were characterized through the application of diverse techniques (XRPD, FT-IR spectroscopy, SEM, EDS, and TEM microscopy, thermogravimetric analysis, and magnetization measurements), while ecotoxicity was assessed through a standard test on the freshwater organism Daphnia magna. Independently of the synthesis procedure, the magnetic composites were successfully obtained. The Fe3O4 is nanometric (about 10 nm) and the weight percentage is sample-dependent. It decorates the HNT's surface and also forms aggregates linking the nanotubes in Fe3O4-rich samples. Thermodynamic and kinetic experiments showed different adsorption capacities of OFL, ranging from 23 to 45 mg g-1. The kinetic process occurred within a few minutes, independently of the composite. The capability of the three HNT/Fe3O4 in removing the OFL was confirmed under realistic conditions, when OFL was added to tap, river, and effluent waters at µg L-1 concentration. No acute toxicity of the composites was observed on freshwater organisms. Despite the good results obtained for all the composites, the sample by co-precipitation is the most performant as it: (i) is easily magnetically separated from the media after the use; (ii) does not undergo any degradation after three adsorption cycles; (iii) is synthetized through a low-cost procedure. These features make this material an excellent candidate for removal of OFL from water.

Microplastic contamination of supraglacial debris differs among glaciers with different anthropic pressures.

Microplastic (MP) contamination is ubiquitous and widespread in terrestrial and aquatic ecosystems, including remote areas. However, information on the presence and distribution of MPs in high-mountain ecosystems, including glaciers, is still limited. The present study aimed at investigating presence, spatial distribution, and patterns of contamination of MPs on three glaciers of the Ortles-Cevedale massif (Central Alps, Northern Italy) with different anthropic pressures, i.e., the Forni, Cedec and Ebenferner-Vedretta Piana glaciers. Samples of supraglacial debris were randomly collected from the glaciers and MPs were isolated. The mean amount (±SE) of MPs measured in debris from Forni, Cedec and Ebenferner-Vedretta Piana glaciers was 0.033 ± 0.007, 0.025 ± 0.009, and 0.265 ± 0.027 MPs g-1 dry weight, respectively. The level and pattern of MP contamination from the Ebenferner-Vedretta Piana glacier were significantly different from those of the other glaciers. No significant spatial gradient in MP distribution along the ablation areas of the glaciers was observed, suggesting that MPs do not accumulate toward the glacier snout. Our results confirmed that local contamination can represent a relevant source of MPs in glacier ecosystems experiencing high anthropic pressure, while long-range transport can be the main source on other glaciers.

The hidden cost of following currents: Microplastic ingestion in a planktivorous seabird.

Microplastics are increasingly pervasive pollutants, particularly abundant in the neuston where they drift with currents. We assessed dietary microplastic ingestion in the Mediterranean storm petrel (Hydrobates pelagicus melitensis), a small pelagic seabird that forages on plankton and inhabit the Mediterranean sea, one of the most polluted seas worldwide. We collected spontaneous regurgitates from 30 chick-rearing individuals and used GPS tracking data from 7 additional individuals to locate foraging areas. Birds foraged in pelagic areas characterized by water stirring and mixing, and regurgitates from 14 individuals (i.e. 45 %) contained microplastics. Fibers were the dominant shape (56 %), with polyester, polyethylene and nylon being the most frequent polymers. Our findings highlight the potential sensitivity of this species of conservation interest to plastic pollution and suggest that storm petrel regurgitates can be a valuable matrix to investigate microplastic ingestion in planktonic foragers, providing a characterization of spatio-temporal patterns of microplastic exposure in pelagic environments.

Approaches for Management and Valorization of Non-Homogeneous, Non-Recyclable Plastic Waste.

The environmental accumulation of plastic wastes has become one of the most discussed topics in the scientific community. The development of new strategies to tackle this issue is of crucial importance, and different approaches are being investigated to effectively reduce plastic waste generated by improper or inefficient disposal. In addition to the efforts addressing the development of biodegradable plastics, the research is currently focused on the development of innovative recycling approaches. Indeed, although most plastic materials are potentially recyclable, only 15% of the worldwide plastic waste is currently recycled, while the remaining 85% is usually incinerated to recover thermal energy or landfilled. The hurdles to efficient recycling come from improper management of end-of-life plastic goods. Moreover, the highly heterogeneous nature and versatility of plastic and polymeric materials have led to the development of multilayered materials, composites, blends and many other different species, whose management and/or reprocessing to yield high-value products is extremely challenging. Thus, although these materials are extremely valuable from an industrial point of view, they add a high degree of complexity to the recycling process because each one of them is different from the other, but they cannot be separated efficiently. The aim of the present review is to return a comprehensive overview of environmental and management issues related to the complex and heterogeneous mixture of plastic waste that is generated at the end of the sorting procedures in Italian plastic recycling plants, the so-called 'Plasmix'. This review lists the difficulties and limitations related to the management of non-recyclable Plasmix and highlights the strategies for the proper, sustainable and valuable use of this plastic waste.

Differential biochemical and behavioral responses induced by cocaine and benzoylecgonine exposure to the red swamp crayfish Procambarus clarkii.

Cocaine (COC) and its main metabolite, the benzoylecgonine (BE), are the main illicit drugs measured in aquatic system worldwide, with concentrations up to hundreds of ng/L. Although their current environmental concentrations are low these molecules can induce adverse effects at sub-individual level in non-target organisms. In contrast, the information at individual and behavioral level are still scant. The present study aimed at investigating biochemical and behavioral effects induced by 14-days exposure to environmentally relevant concentrations (50 ng/L and 500 ng/L) of COC and BE towards Procambarus clarkii. At sub-individual level, the activity of antioxidant and detoxifying (superoxide dismutase - SOD, catalase - CAT, glutathione peroxidase - GPx and glutathione S-transferases - GST) enzymes, as well as the levels of lipid peroxidation (LPO), were measured as oxidative stress-related endpoints. We also measured the acetylcholinesterase (AChE) activity to check for neurotoxic effect of COC and BE. At individual level, the modulation of some behavioral tasks (i.e., response to external stimuli, changes in feeding activity and exploration of a new environment) were assessed. Although both COC and BE exposure did not induce an oxidative stress situation, a significant inhibition of AChE activity was noted, resulting in behavioral changes in crayfish exposed to COC only. Crayfish exposed to the higher COC concentration showed an increase in the boldness and a decrease in the feeding activity, suggesting that COC may act according to its psychotropic mode of action.

Isolation and evaluation of quinoa (Chenopodium quinoa Willd.) protein fractions. A nutritional and bio-functional approach to the globulin fraction.

This study evaluated the solubility profiles of quinoa grain proteins and applied a complete process for the isolation of its main protein fractions, namely: albumins, globulins, prolamins and glutelins, which corresponded to 26.96%, 41.3%, 1.7% and 23.16% of the total protein content, respectively. When these fractions were digested with pepsin followed by pancreatin, the degrees of hydrolysis achieved varied between 26.62% (for unheated globulin fraction) and 38.97% (for unheated glutelin), with casein reached 33.73% hydrolysis. After heating, the globulin hydrolysis degree increased to 34.7%, not significantly differing from casein. These results reflect its good susceptibility to hydrolysis by digestive enzymes, and this observation is reinforced with assays with pepsin, trypsin and chymotrypsin tested separately. Globulins, the largest protein fraction, showed promising results in additional assays regarding the amino acid profile, with limitation only for lysine in relation to the FAO standard, and the potential for releasing bioactive peptides after digestion. Although pepsin-digested globulin inhibited only 5% of ACE activity under the conditions tested, after 24h with the addition of pancreatin, the inhibition was 100%. Antioxidant activity (DPPH assay) also indicated very similar results, when hydrolysis with pepsin was inefficient in releasing antioxidant peptides, while hydrolysis by pancreatin led to 35 times greater results.

Zinc oxide, titanium dioxide and C60 fullerene nanoparticles, alone and in mixture, differently affect biomarker responses and proteome in the clam Ruditapes philippinarum.

Continuous release of nanoparticles (NPs) into marine coastal environments results in an increased risk of exposure to complex NP mixtures for marine organisms. However, to date, the information on the effects at molecular and biochemical levels induced by the exposure to NPs, singly and as a mixture, is still scant. The present work aimed at exploring the independent and combined effects and the mechanism(s) of action induced by 7-days exposure to 1 µg/L nZnO, 1 µg/L nTiO2 and 1 µg/L FC60 fullerene in the Manila clam Ruditapes philippinarum, using a battery of immunological and oxidative stress biomarkers in haemolymph, gills and digestive gland. In addition, proteomics analyses were performed in gills and the digestive gland, where NP bioaccumulation was also assessed. Increased bioaccumulation of single NPs and the mixture was linked with increased oxidative stress and higher damage to proteins, lipids and DNA in all tissues analysed. The proteomics approach highlighted protein modulation in terms of abundance and damage (higher redox-thiol and carbonylated groups content). In particular, the modulated proteins (16 in gills and 18 in digestive gland) were mostly related to cytoskeleton and energetic metabolism. The digestive gland was the tissue more affected. For all biomarkers measured, increased detrimental effects were observed in the mixture compared to single NP exposures.