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Larval settlement is a critical step for sessile benthic species such as corals, whose ability to thrive on diverse natural and anthropogenic substrates may lead to a competitive advantage in the colonization of new environments with respect to a narrow tolerance for a specific kind of substratum. Plastic debris, widespread in marine waters, provides a large, motile, and solid substratum supporting a highly diverse biological community. Here we present the first observation of a floating plastic bottle colonized by the deep-sea coral Desmophyllum dianthus. The density pattern and co-occurring species composition suggest a pioneer behavior of this coral species, whose peculiar morphologic plasticity response when interacting with the plastic substrate (i.e., low density polyethylene) has not been observed before. The tolerance of D. dianthus for such plastic substrate may affect ecological processes in deep water environments, disrupting interspecific substrate competition in the benthic community.

The sea urchin Paracentrotus lividus (P. lividus) was exposed to either virgin or biofilm-covered polystyrene microbeads (micro-PS, 45 μm) in order to test the effect of microbial colonization on the uptake, biodistribution, and immune response. The biofilm was dominated by bacteria, as detected by scanning electron microscopy and 16S rRNA sequencing. A higher internalization rate of colonized micro-PS inside sea urchins compared to virgin ones was detected, suggesting a role of the plastisphere in the interaction. Colonized and virgin micro-PS showed the same biodistribution pattern by accumulating mainly in the digestive system with higher levels and faster egestion rates for the colonized. However, a significant increase of catalase and total antioxidant activity was observed only in the digestive system of colonized micro-PS-exposed individuals. Colonized micro-PS also induced a significant decrease in the number of coelomocytes with a significant increase in vibratile cells, compared to control and virgin micro-PS-exposed animals. Moreover, a general time-dependent increase in the red/white amoebocytes ratio and reactive oxygen species and a decrease in nitrogen ones were observed upon exposure to both colonized and virgin micro-PS. Overall, micro-PS colonization clearly affected the uptake and toxicological responses of the Mediterranean sea urchin P. lividus in comparison to virgin micro-PS.

Nanoplastics are considered contaminants of emerging concern at the global scale. The recent evidence of their occurrence in seawater from the Mediterranean Sea calls for a thorough evaluation of their impact on marine life and in particular on vulnerable life stages such as planktonic embryos. Here, we investigated the impact of increasing nominal concentrations of 50 nm amino-modified (PS-NH

Marine nano-ecotoxicology has emerged with the purpose to assess the environmental risks associated with engineered nanomaterials (ENMs) among contaminants of emerging concerns entering the marine environment. ENMs’ massive production and integration in everyday life applications, associated with their peculiar physical chemical features, including high biological reactivity, have imposed a pressing need to shed light on risk for humans and the environment. Environmental safety assessment, known as ecosafety, has thus become mandatory with the perspective to develop a more holistic exposure scenario and understand biological effects. Here, we review the current knowledge on behavior and impact of ENMs which end up in the marine environment. A focus on titanium dioxide (n-TiO2) and silver nanoparticles (AgNPs), among metal-based ENMs massively used in commercial products, and polymeric NPs as polystyrene (PS), largely adopted as proxy for nanoplastics, is made. ENMs eco-interactions with chemical molecules including (bio)natural ones and anthropogenic pollutants, forming eco- and bio-coronas and link with their uptake and toxicity in marine organisms are discussed. An ecologically based design strategy (eco-design) is proposed to support the development of new ENMs, including those for environmental applications (e.g., nanoremediation), by balancing their effectiveness with no associated risk for marine organisms and humans.

Hydrothermal carbonization (HTC) of sewage sludge reduces the waste volume and can be source of energy and valuable products. Furthermore, HTC offers several advantages over conventional dry-thermal pre-treatments, as no prior drying is requested, and the high quality of the char produced promotes applications as energy production and storage, wastewater remediation, and soil amendment. Relationships between char yields, physicochemical properties and process parameters are here analysed, with the aim to provide insight into the choice of the process severity required to fit the desired application. Moreover, presence and fate of heavy metals and organic contaminants are discussed. The highest reaction temperature is the main parameter affecting the physicochemical characteristics of the char produced, while the heating rate governs the heat mass transfer and the rate of intermediates formation. Depolymerization of the biomass results in a reduction of the oxygen to carbon ratio and, therefore, in augmented high heating values, further increased by deposition of 5-(hydroxymethyl)furfural. Recirculation of process water may enhance dehydration reactions and the deposition of degraded polymers, increasing dewaterability and yield, but field trials are recommended to assess the feasibility of this option. An overuse of chars for energy generation purposes would be deleterious for the environmental life cycle. Further research is encouraged to assess the pollutants abatement and their degradation pathways when incorporated in the carbonaceous product, to promote the application of hydrochars as soil amendment, as well as for environmental remediation purposes.

Plastic debris has been recognised as a potential stressor for Antarctic marine organisms. In this study, the effects of surface charged polystyrene nanoparticles (PS NPs) on the immune cells (coelomocytes) of the Antarctic sea urchin Sterechinus neumayeri were assessed through in vitro short-term cultures. The behaviour of anionic carboxylated (PS-COOH) and cationic amino-modified (PS-NH2) NPs in filtered natural sea water (NSW) from King George Island (South Shetland Islands) was characterised by dynamic light scattering. Cellular morphology, NP uptake, phagocytic capacity and gene expression were evaluated after 6 and 24 h of exposure to 1 and 5 µg mL−1 PS NPs. Secondary characterisation showed an initial good dispersion of PS NPs in NSW, followed by nano-scale aggregation after 24 h. Both PS NPs affected cellular phagocytosis and generated an inflammatory response against oxidative stress and apoptosis at the molecular level. Fluorescently labelled PS-COOH aggregates were internalised by phagocytes and associated to the modulation of genes related to external challenges, antioxidant responses and cell protection against stress and apoptosis. Exposure to PS-NH2 caused a strong decrease in phagocytic capacity and the formation of cellular debris at 5 µg mL−1 after 24 h, but low gene modulation, suggesting a threshold in coelomocytes defence ability against PS-NH2. This study represents the first attempt to assess the impact of nanoplastics on Antarctic marine organisms. Our findings demonstrate that PS NPs with different surface charges constitute a challenge for S. neumayeri immune cells.

In recent years, the application of engineered nanomaterials (ENMs) in environmental remediation gained increasing attention. Due to their large surface area and high reactivity, ENMs offer the potential for the efficient removal of pollutants from environmental matrices with better performances compared to conventional techniques. However, their fate and safety upon environmental application, which can be associated with their release into the environment, are largely unknown. It is essential to develop systems that can predict ENM interactions with biological systems, their overall environmental and human health impact. Until now, Life-Cycle Assessment (LCA) tools have been employed to investigate ENMs potential environmental impact, from raw material production, design and to their final disposal. However, LCA studies focused on the environmental impact of the production phase lacking information on their environmental impact deriving from in situ employment. A recently developed eco-design framework aimed to fill this knowledge gap by using ecotoxicological tools that allow the assessment of potential hazards posed by ENMs to natural ecosystems and wildlife. In the present review, we illustrate the development of the eco-design framework and review the application of ecotoxicology as a valuable strategy to develop ecosafe ENMs for environmental remediation. Furthermore, we critically describe the currently available ENMs for marine environment remediation and discuss their pros and cons in safe environmental applications together with the need to balance benefits and risks promoting an environmentally safe nanoremediation (ecosafe) for the future.

Benzo(a)pyrene (B(a)P) is a well-known genotoxic agent, the removal of which from environmental matrices is mandatory, necessitating the application of cleaning strategies that are harmless to human and environmental health. The potential application of nanoparticles (NPs) in the remediation of polluted environments is of increasing interest. Here, specifically designed NPs were selected as being non-genotoxic and able to interact with B(a)P, in order to address the genetic and chromosomal damage it produces. A newly formulated pure anatase nano-titanium (nano-TiO2), a commercial mixture of rutile and anatase, and carbon black-derived hydrophilic NPs (HNP) were applied. Once it had been ascertained that the NPs selected for the work did not induce genotoxicity, marine mussel gill biopsies were exposed in vitro to B(a)P (2 μg/mL), alone and in combination with the selected NPs (50 µg/mL nano-TiO2, 10 µg/mL HNP). DNA primary reversible damage was evaluated by means of the Comet assay. Chromosomal persistent damage was assessed on the basis of micronuclei frequency and nuclear abnormalities by means of the Micronucleus-Cytome assay. Transmission Electron Microscopy (TEM) was performed to investigate the mechanism of action exerted by NPs. Pure Anatase n-TiO2 was found to be the most suitable for our purpose, as it is cyto- and genotoxicity free and able to reduce the genetic and chromosomal damage associated with exposure to B(a)P.

In Antarctic regions, the composition and metabolic activity of microbial assemblages associated with plastic debris (“plastisphere”) are almost unknown. A macroplastic item from land (MaL, 30 cm) and a mesoplastic from the sea (MeS, 4 mm) were collected in Maxwell Bay (King George Island, South Shetland) and analyzed by Fourier transform infrared spectroscopy in attenuated total reflectance geometry (FTIR-ATR), which confirmed a polystyrene foam and a composite high-density polyethylene composition for MaL and MeS, respectively. The structure and function of the two plastic-associated prokaryotic communities were studied by complementary 16S ribosomal RNA gene clone libraries, total bacterioplankton and culturable heterotrophic bacterial counts, enzymatic activities of the whole community and enzymatic profiles of bacterial isolates. Results showed that Gamma- and Betaproteobacteria (31% and 28%, respectively) dominated in MeS, while Beta- and Alphaproteobacteria (21% and 13%, respectively) in MaL. Sequences related to oil degrading bacteria (Alcanivorax,Marinobacter) confirmed the known anthropogenic pressure in King George Island. This investigation on plastic-associated prokaryotic structure and function represents the first attempt to characterize the ecological role of plastisphere in this Antarctic region and provides the necessary background for future research on the significance of polymer type, surface characteristics and environmental conditions in shaping the plastisphere.

The increasing use and disposal of plastics has become a persistent problem in the marine environment, calling for studies that refer to realistic scenarios to understand their effects on biota. Particularly, the understanding about the effects of co-exposure with nanoplastic particles and metals on aquatic organisms is still limited. The present work aimed to investigate the acute toxicity of amino-functionalized polystyrene nanoparticles (PS-NH2; 50 nm) as proxy for nanoplastics on brine shrimp Artemia franciscana larvae under different culture conditions and at different stages of development, as well as the combined effect with two reference toxicants - potassium dichromate (K2Cr2O7) and copper sulfate (CuSO4). Nauplii (instar II or III larval stages) were exposed to different concentrations of PS-NH2 (0.005 to 5 μg mL-1) for up to 48 h, with or without agitation in order to mimic a more realistic environmental scenario. Larval mobility and PS-NH2 accumulation were monitored under microscopy. PS-NH2 alone showed toxicity only at the highest concentration tested (5 μg mL-1) regardless the incubation method used (61.2 + 3.1% and 65.0 + 4.5% with and without agitation, respectively). Moreover, instar III stage was the most sensitive to PS-NH2 exposure (38.2% immobility in 24 h of exposure; 5 μg mL-1). Evidence of PS-NH2 retention in the gastrointestinal tract in a concentration- and time-dependent manner was also obtained. Mixtures of PS-NH2 (0.005 and 5 μg mL-1) with different concentrations of K2Cr2O7 increased the immobilization rate of the larvae after 48 h of exposure, when compared to the K2Cr2O7 alone. Similar results were observed for CuSO4 in the co-exposure conditions at different concentrations. However, exposing nauplii to a mixture of PS-NH2 (0.005 μg mL-1) and CuSO4 decreased immobilization rate, in comparison to the group exposed to CuSO4 alone. The present work highlights the potential risk posed by nanoplastics to zooplanktonic species through their interaction with other toxicants.

Plastic debris has been identified as a potential threat to Antarctic marine ecosystems, however, the impact of nanoplastics (

To encourage the applicability of nano-adsorbent materials for heavy metal ion removal from seawater and limit any potential side effects for marine organisms, an ecotoxicological evaluation based on a biological effect-based approach is presented. ZnCl2 (10 mg L&minus, 1) contaminated artificial seawater (ASW) was treated with newly developed eco-friendly cellulose-based nanosponges (CNS) (1.25 g L&minus, 1 for 2 h), and the cellular and tissue responses of marine mussel Mytilus galloprovincialis were measured before and after CNS treatment. A control group (ASW only) and a negative control group (CNS in ASW) were also tested. Methods: A significant recovery of Zn-induced damages in circulating immune and gill cells and mantle edges was observed in mussels exposed after CNS treatment. Genetic and chromosomal damages reversed to control levels in mussels&rsquo, gill cells (DNA integrity level, nuclear abnormalities and apoptotic cells) and hemocytes (micronuclei), in which a recovery of lysosomal membrane stability (LMS) was also observed. Damage to syphons, loss of cilia by mantle edge epithelial cells and an increase in mucous cells in ZnCl2-exposed mussels were absent in specimens after CNS treatment, in which the mantle histology resembled that of the controls. No effects were observed in mussels exposed to CNS alone. As further proof of CNS&rsquo, ability to remove Zn(II) from ASW, a significant reduction of &gt, 90% of Zn levels in ASW after CNS treatment was observed (from 6.006 to 0.510 mg L&minus, 1). Ecotoxicological evaluation confirmed the ability of CNS to remove Zn from ASW by showing a full recovery of Zn-induced toxicological responses to the levels of mussels exposed to ASW only (controls). An effect-based approach was thus proven to be useful in order to further support the environmentally safe (ecosafety) application of CNS for heavy metal removal from seawater.

Titanium Dioxide Nanoparticles (TiO2 NPs) could cause alterations in exposed aquatic species, in terms of oxygen consumption, ammonia excretion and tissues functionality therefore, the aim of the present study is to evaluate the effects of acute exposure to different concentrations of TiO2 NPs (0.1, 0.5, 1.0 and 2.5 mgL-1) on routine metabolism (oxygen consumption and ammonia excretion) and histological parameters (branchial and hepatic) in Oreochromis niloticus. After 24 hours, we observed an increase in oxygen consumption of 2.36 and 3.23 times in groups exposed to 1.0 and 2.5 mgL-1 of TiO2 NPs respectively, as well as an increase in ammonia excretion of 3.54, 4.0 and 4.82 times higher in groups exposed to 0.5, 1.0 and 2.5 mgL-1 of TiO2 NPs respectively, compared to the control group. Histological analysis showed, after 72 hours, moderate to severe alterations in both gills and liver of TiO2 exposed fish at concentrations 1.0 and 2.5 mgL-1, the severity and occurrence of the alteration observed was grade 3 (severe and extensive pathological alterations). We concluded that waterborne exposure of Nile tilapia to TiO2 NPs caused alteration in routine metabolism and histological parameters in a dose-dependent manner. Nanoparticulas de Dioxido de Titânio (TiO2NPs) podem causar alteracoes nas especies aquaticas expostas, em termos de consumo de oxigenio, excrecao de amonia e funcionalidade de tecidos, portanto, o objetivo do presente estudo e avaliar os efeitos da exposicao aguda a diferentes concentracoes de TiO2NPs (0,1, 0,5, 1,0 e 2,5 mgL-1) sobre o metabolismo de rotina (consumo de oxigenio e excrecao de amonia) e parâmetros histologicos (branquiais e hepaticos) em Oreochromis niloticus. Apos 24 horas, observamos um aumento no consumo de oxigenio de 2,36 e 3,23 vezes em grupos expostos a 1,0 e 2,5 mgL-1 de TiO2NPs respectivamente, bem como um aumento na excrecao de amonia de 3,54, 4,0 e 4,82 vezes maior nos grupos expostos a 0,5, 1,0 e 2,5 mgL-1 de TiO2NPs respectivamente, em comparacao com o grupo controle. A analise histologica mostrou, apos 72 horas, alteracoes moderadas a severas tanto nas brânquias quanto no figado de peixes expostos a TiO2NPs em concentracoes de 1,0 e 2,5 mgL-1, a gravidade e a ocorrencia da alteracao observada foram de grau 3 (alteracoes patologicas graves e extensas). Concluimos que a exposicao a TiO2NPs da tilapia do Nilo causaram alteracao no metabolismo de rotina e nos parâmetros histologicos de uma maneira dose-dependente.

Titanium dioxide nanoparticles (TiO 2 NPs) have been widely employed in industrial applications, thus rising concern about their impact in the aquatic environment. In this study we investigated the chemical behaviour of TiO 2 NPs in the culture medium and its effect on the green alga Dunaliella tertiolecta , in terms of growth inhibition, oxidative stress, ROS (Reactive Oxygen Species) accumulation and chlorophyll content. In addition, the influence of exopolymeric substances (EPS) excreted by the microalgae on the stability of NPs has been evaluated. The physicochemical characterization showed a high propensity of TiO 2 NPs to form micrometric-sized aggregates within 30 min, large enough to partially settle to the bottom of the test vessel. Indeed, an increasing amount of TiO 2 particles settled out with time, but the presence of EPS seemed to mitigate this behaviour in the first 6 h of exposure where the main effects in D. tertiolecta were observed. TiO 2 NPs did not inhibit the 72-h growth rate of D. tertiolecta , nor affected the cellular chlorophyll concentration in the range 0.01–10 mg L −1 . The time-course of ROS production showed an initial transient increase of ROS in TiO 2 NP-exposed algae compared to the control, concomitant with an enhancement of catalase activity. Interestingly, intracellular ROS was a small fraction of total ROS, the highest amount being extracellular. The occurrence of cell-mediated chemical transformations of TiO 2 NPs in the external medium, related to the presence of EPS, has been evaluated. Our results showed that carbohydrates were the major component of EPS, whereas proteins of medium molecular weight (20–80 kDa) were preferentially bound to TiO 2 NPs, likely influencing their biological fate.

The Gulf of Follonica (Italy) is impacted by the chemical pollution from ancient mining activity and present industrial processes. This study was aimed to determine the bioavailability of dioxin-like compounds (DLCs) in coastal marine environment and to assess the genotoxic potential of waste waters entering the sea from an industrial canal. Moderately high levels of DCLs compounds (∑ PCDDs + PCDFs 2.18–29.00 pg/g dry wt) were detected in Mytilus galloprovincialis transplanted near the waste waters canal and their corresponding Toxic Equivalents (TEQs) calculated. In situ exposed mussels did not show any genotoxic effect (by Comet and Micronucleus assay). Otherwise, laboratory exposure to canal waters exhibited a reduced genomic template stability (by RAPD-PCR assay) but not DNA or chromosomal damage. Our data reveal the need to focus on the levels and distribution of DLCs in edible species from the study area considering their potential transfer to humans through the consumption of sea food.

The aim of the present study was to assess the occurrence and spatial distribution of PCDD/Fs and dioxin-like compounds in topsoils of Taranto (Apulia Region), one of the most heavily industrialized and contaminated area of Southern Italy. A combined approach of chemical analysis by GC-MS/MS and AhR reporter gene bioassay was applied in a subset of topsoil samples (n = 20) collected in 2017-18 from ten sites embracing three levels of risk (from high to low) in the framework of a large survey inside Taranto municipality. TCDD-BEQs and GC-MS/MS TEQWHO and TEQTHEORETICAL revealed a decreasing trend with the distance from main industrial settings and landfill areas. A strong correlation between TCDD-BEQs and TEQWHO values (R2 = 0.85) and TEQTHEORETICAL (R2 = 0.88) was also found. In 3 out of 10 topsoil investigated, BEQs and TEQWHO/THEORETICAL resulted above Italian National Regulatory Limits for ∑PCDD/Fs in green, private and recreational used soils (10 ng TEQ/kg d.w. D.Lgs 152/2006) and for ∑PCDD/F/dl-PCBs in agricultural and farming soil (6 ng TEQ/kg d.w. D.M. 46/2019). GC-MS/MS pattern revealed the highest prevalence of dl-PCBs in 6 out of 10 sites, followed by PCDFs and PCDDs. Those sites are all located in proximity of main industrial steel and iron ore sinter plant, steel plant’s landfills and illegal dumping sites. An update on occurrence and spatial distribution of PCDD/Fs and dl-PCBs contamination of Taranto urban soils was obtained and the DR-CALUX® bioassay was further recommended as a suitable screening tool for environmental and human risk assessment.

Plastic pollution has been globally recognized as a critical issue for marine ecosystems and nanoplastics constitute one of the last unexplored areas to understand the magnitude of this threat. However, current difficulties in sampling and identifying nano-sized debris make hard to assess their occurrence in marine environment. Polystyrene nanoparticles (PS NPs) are largely used as nanoplastics in ecotoxicological studies and although acute exposures have been already investigated, long-term toxicity on marine organisms is unknown. Our study aims at evaluating the effects of 40nm PS anionic carboxylated (PS-COOH) and 50nm cationic amino-modified (PS-NH2) NPs in two planktonic species, the green microalga Dunaliella tertiolecta and the brine shrimp Artemia franciscana, respectively prey and predator. PS NP behaviour in exposure media was determined through DLS, while their toxicity to microalgae and brine shrimps evaluated through 72h growth inhibition test and 14 d long-term toxicity test respectively. Moreover, the expression of target genes (i.e. clap and cstb), having a role in brine shrimp larval growth and molting, was measured in 48h brine shrimp larvae. A different behaviour of the two PS NPs in exposure media as well as diverse toxicity to the two planktonic species was observed. PS-COOH formed micro-scale aggregates (Z-Average>1μm) and did not affect the growth of microalgae up to 50μg/ml or that of brine shrimps up to 10μg/ml. However, these negatively charged NPs were adsorbed on microalgae and accumulated (and excreted) in brine shrimps, suggesting a potential trophic transfer from prey to predator. On the opposite, PS-NH2-formed nano-scale aggregates (Z-Average

Magnetic hydrogels based on carboxymethylcellulose (CMC) were prepared using magnetite (Fe 3 O 4 ) nanoparticles (NPs) functionalized with 3-aminopropyltrimethoxysilane (APTMS) as cross-linker. In this hybrid organic-inorganic hydrogel, magnetite NPs are covalently bonded to the polymer chains. The CMC-Fe 3 O 4 hydrogel was characterized by means of Attenuated Total Reflection Fourier Transform Infra Red Spectroscopy (ATR-FTIR) and Field Emission Scanning Electron Microscopy (FESEM). The equilibrium swelling degree of the CMC-Fe 3 O 4 hydrogels was measured in deionized water as well as in artificial and natural sea water. Bioassays were performed in order to assess the ecosafety of these magnetic hydrogels by testing any potential effect on the growth rate of marine microalgae Dunaliella tertiolecta . The capability of the CMC-Fe 3 O 4 hydrogel to adsorb organic and inorganic pollutants was tested on solutions of methylene blue (MB) and CdCl 2 in deionized and sea water as model systems. The CMC-Fe 3 O 4 hydrogel is capable to adsorb both MB and Cd(II) in deionized water. On the other hand, the adsorption capacity is strongly reduced in sea water, due to the ions in solution which decrease the electrostatic attraction between the carboxylate groups of CMC and the positive charge of MB and Cd(II).

Plastics are the most abundant marine debris globally dispersed in the oceans and its production is rising with documented negative impacts in marine ecosystems. However, the chemical-physical and biological interactions occurring between plastic and planktonic communities of different types of microorganisms are poorly understood. In these respects, it is of paramount importance to understand, on a molecular level on the surface, what happens to plastic fragments when dispersed in the ocean and directly interacting with phytoplankton assemblages. This study presents a computer-aided analysis of electron paramagnetic resonance (EPR) spectra of selected spin probes able to enter the phyoplanktonic cell interface and interact with the plastic surface. Two different marine phytoplankton species were analyzed, such as the diatom Skeletonema marinoi and dinoflagellate Lingulodinium polyedrum, in absence and presence of polyethylene terephthalate (PET) fragments in synthetic seawater (ASPM), in order to in-situ characterize the interactions occurring between the microalgal cells and plastic surfaces. The analysis was performed at increasing incubation times. The cellular growth and adhesion rates of microalgae in batch culture medium and on the plastic fragments were also evaluated. The data agreed with the EPR results, which showed a significant difference in terms of surface properties between the diatom and dinoflagellate species. Low-polar interactions of lipid aggregates with the plastic surface sites were mainly responsible for the cell-plastic adhesion by S. marinoi, which is exponentially growing on the plastic surface over the incubation time.

Silver nanoparticles (AgNPs) are widely used as engineered nanomaterials (ENMs) in many advanced nanotechnologies, due to their versatile, easy and cheap preparations combined with peculiar chemical-physical properties. Their increased production and integration in environmental applications including water treatment raise concerns for their impact on humans and the environment. An eco-design strategy that makes it possible to combine the best material performances with no risk for the natural ecosystems and living beings has been recently proposed. This review envisages potential hybrid solutions of AgNPs for water pollution monitoring and remediation to satisfy their successful, environmentally safe (ecosafe) application. Being extremely efficient in pollutants sensing and degradation, their ecosafe application can be achieved in combination with polymeric-based materials, especially with cellulose, by following an eco-design approach. In fact, (AgNPs)–cellulose hybrids have the double advantage of being easily produced using recycled material, with low costs and possible reuse, and of being ecosafe, if properly designed. An updated view of the use and prospects of these advanced hybrids AgNP-based materials is provided, which will surely speed their environmental application with consequent significant economic and environmental impact.

The contamination of freshwaters by heavy metals represents a great problem, posing a threat for human and environmental health. Cadmium is classified as carcinogen to humans and its mechanism of carcinogenicity includes genotoxic events. In this study a recently developed eco-friendly cellulose-based nanosponge (CNS) was investigated as a candidate in freshwater nano-remediation process. For this purpose, CdCl2 (0.05 mg L&minus, 1) contaminated artificial freshwater (AFW) was treated with CNS (1.25 g L&minus, 1 for 2 h), and cellular responses were analyzed before and after CNS treatment in Dreissena polymorpha hemocytes. A control group (AFW) and a negative control group (CNS in AFW) were also tested. DNA primary damage was evaluated by Comet assay while chromosomal damage and cell proliferation were assessed by Cytome assay. AFW exposed to CNS did not cause any genotoxic effect in zebra mussel hemocytes. Moreover, DNA damage and cell proliferation induced by Cd(II) turned down to control level after 2 days when CNS were used. A reduction of Cd(II)-induced micronuclei and nuclear abnormalities was also observed. CNS was thus found to be a safe and effective candidate in cadmium remediation process being efficient in metal sequestering, restoring cellular damage exerted by Cd(II) exposure, without altering cellular physiological activity.

Environment-friendly nanosponges, having a high content of carboxyl groups, were synthesized by crosslinking &beta, cyclodextrin and linecaps, a highly soluble pea starch derivative, with citric acid in water. Additionally, pyromellitic nanosponges were prepared by reacting &beta, cyclodextrin and linecaps with pyromellitic dianhydride in dimethyl sulfoxide and used in comparison with the citric nanosponges. After ion-exchange of the carboxyl groups H+ with sodium ions, the ability of the nanosponges to sequester heavy metal cations was investigated. At a metal concentration of 500 ppm, the pyromellitate nanosponges exhibited a higher retention capacity than the citrate nanosponges. At lower metal concentrations (&le, 50 ppm) both the citrate and the pyromellitate nanosponges showed high retention capacities (up to 94% of the total amount of metal), while, in the presence of interfering sea water salts, the citrate nanosponges were able to selectively adsorb a significantly higher amount of heavy metals than the pyromellitate nanosponges, almost double in the case of Cu2+.

Marine diatoms have been identified among the most abundant taxa of microorganisms associated with plastic waste collected at sea. However, the impact of nano-sized plastic fragments (nanoplastics) at single cell and population level is almost unknown. We exposed the marine diatom Skeletonema marinoi to model polystyrene nanoparticles with carboxylic acid groups (PS–COOH NPs, 90 nm) for 15 days (1, 10, 50 μg/mL). Growth, reactive oxygen species (ROS) production, and nano-bio-interactions were investigated. No effect on diatom growth was observed, however Dynamic light scattering (DLS) demonstrated the formation of large PS aggregates which were localized at the diatoms’ fultoportula process (FPP), as shown by TEM images. Increase production of ROS and reduction in chain length were also observed upon PS NPs exposure (p < 0.005). The observed PS-diatom interaction could have serious consequences on diatoms ecological role on the biogeochemical cycle of carbon, by impairing the formation of fast-sinking aggregates responsible for atmospheric carbon fixation and sequestration in the ocean sea floor. S. marinoi exposure to PS NPs caused an increase of intracellular and extracellular oxidative stress, the reduction of diatom's chain length and the adhesion of PS NPs onto the algal surface.

In this work, hydrophilic silver nanoparticles (AgNPs), bifunctionalized with citrate (Cit) and L-cysteine (L-cys), were synthesized. The typical local surface plasmon resonance (LSPR) at &lambda, max = 400 nm together with Dynamic Light Scattering (DLS) measurements (&lt, 2RH&gt, = 8 &plusmn, 1 nm) and TEM studies (&Oslash, = 5 &plusmn, 2 nm) confirmed the system nanodimension and the stability in water. Molecular and electronic structures of AgNPs were investigated by FTIR, SR-XPS, and NEXAFS techniques. We tested the system as plasmonic sensor in water with 16 different metal ions, finding sensitivity to Hg2+ in the range 1&ndash, 10 ppm. After this first screening, the molecular and electronic structure of the AgNPs-Hg2+ conjugated system was deeply investigated by SR-XPS. Moreover, in view of AgNPs application as sensors in real water systems, environmental safety assessment (ecosafety) was performed by using standardized ecotoxicity bioassay as algal growth inhibition tests (OECD 201, ISO 10253:2006), coupled with determination of Ag+ release from the nanoparticles in fresh and marine aqueous exposure media, by means of ICP-MS. These latest studies confirmed low toxicity and low Ag+ release. Therefore, these ecosafe AgNPs demonstrate a great potential in selective detection of environmental Hg2+, which may attract a great interest for several biological research fields.

The use of indicator species has become a standard monitoring approach in environmental risk assessment; however, pollutant bioavailability and its geographical distribution as well as species-sensitiveness could be affected by different factors. The present study investigated the potential use of the land snail Cornu aspersum as bioindicator of industrial air pollution by using cytological and biochemical responses and trace element levels in individuals caged in proximity to an industrial setting. Eleven sites were selected based on wind direction and distance. Snails were placed in holed plastic cages, transplanted to each site and maintained for 30 days fed at libitum and under constant humidity. Several oxidative stress responses including catalase (CAT) and glutathione-s-transferase (GST) activities, levels of malondialdehyde (MDA) and total metallothioneins (MTs) content were determined in snail hepatopancreas while lysosomal membrane stability (LMS) and loss of DNA integrity through micronuclei frequency (MN) were assessed in haemocytes. Trace elements were also analysed in snail whole soft tissues. Multi-model inference used to predict snail’s biological responses in relation to trace elements levels and distance from the industrial setting was able to disentangle the relative contributions of different influencing predictors and could be successfully applied in environmental risk assessment by using land snail as bioindicator species.

There is evidence and serious concern that microplastics have reached the most remote regions of the planet, but how far have they travelled in terrestrial ecosystems? This study presents the first field-based evidence of plastic ingestion by a common and central component of Antarctic terrestrial food webs, the collembolan Cryptopygus antarcticus . A large piece of polystyrene (PS) foam (34 × 31 × 5 cm) covered by microalgae, moss, lichens and microfauna was found in a fellfield along the shores of the Fildes Peninsula (King George Island). The application of an improved enzymatic digestion coupled with Fourier transform infrared microscopy (µ-FTIR), unequivocally detected traces of PS (less than 100 µm) in the gut of the collembolans associated with the PS foam and documented their ability to ingest plastic. Plastics are thus entering the short Antarctic terrestrial food webs and represent a new potential stressor to polar ecosystems already facing climate change and increasing human activities. Future research should explore the effects of plastics on the composition, structure and functions of polar terrestrial biota.

The retrieval of a polystyrene macro-plastic piece stranded on the shores in King George Island (South Shetlands, Antarctica) gave the opportunity to explore the associated bacterial flora. A total of 27 bacterial isolates were identified by molecular 16s rRNA gene sequencing and 7 strains were selected and screened for their ability to produce biofilm and antibiotic susceptibility profiles. All the bacterial isolates were able to produce biofilm. The Kirby-Bauer disk diffusion susceptibility test to 34 antibiotics showed multiple antibiotic resistances against the molecules cefuroxime and cefazolin (belonging to cephalosporins), cinoxacin (belonging to quinolones) and ampicillin, amoxicillin + clavulanic acid, carbenicillin and mezlocillin (belonging to beta-lactams). The obtained results suggest that plastics can serve as vectors for the spread of multiple resistances to antibiotics across Antarctic marine environments and underline the relevance of future studies on this topic.

The adverse effects of engineered nanomaterials (ENM) in marine environments have recently attracted great attention although their effects on marine benthic organisms such as foraminifera are still largely overlooked. Here we document the effects of three negatively charged ENM, different in size and composition, titanium dioxide (TiO2), polystyrene (PS) and silicon dioxide (SiO2), on a microbial eukaryote (the benthic foraminifera Ammonia parkinsoniana) using multiple approaches. This research clearly shows the presence, within the foraminiferal cytoplasm, of metallic (Ti) and organic (PS) ENM that promote physiological stress. Specifically, marked increases in the accumulation of neutral lipids and enhanced reactive oxygen species production occurred in ENM-treated specimens regardless of ENM type. This study indicates that ENM represent ecotoxicological risks for this microbial eukaryote and presents a new model for the neglected marine benthos by which to assess natural exposure scenarios.

Micro- (, This work was financed by Generalitat Valenciana (GV-2014/085-Prometeo-II), and talian Antarctic Research Program (PNRA) contract number: PNRA 16_0075 NANOPANTA: Nano-Polymers in the Antarctic marine environment and biota

Polystyrene nanoparticles (PS NPs) are considered an upcoming threat to marine ecosystems. When NPs encounter biological fluids, they are readily covered by a biomolecular coating, named the “protein corona”, which critically drives the biological fate and toxicity of NPs. Here we provide the first attempt to identify key components of protein coronas of charged amino- and carboxyl-functionalized (PS-NH2 and PS-COOH, respectively) PS NPs exposed to coelomic fluid (CF) of the Mediterranean sea urchin Paracentrotus lividus. The formation of protein coronas conferred identical colloidal features to both positively and negatively charged PS NPs, which acquired a negative ζ potential and monodisperse size distribution in sea urchin CF. Proteomic analysis revealed striking functional analogies of proteins identified from NP–coronas, especially entailing the potential to promote cell association and internalization of both NP types, as mediated by adsorbed protein species. Such basal findings strengthen the importance of the biological identity of NPs in the evaluation of the ecotoxicity potential of polymeric NPs.

Antarctic seabirds well adapted to extreme environments often deal during their life cycle with sub-optimal conditions and occasionally with severe environmental stress. Climate changes, pollution, habitat loss, increasing human presence can all significantly affect organism’s health status from molecular to individual up to population level. In the present study, erythrocyte nuclear abnormalities (ENAs) and white blood cells (WBCs) differential were investigated in 19 adults of Adelie penguin (Pygoscelisadeliae) breeding at Edmonson Point, Antarctic Specially Protected Area (ASPA n. 165) in the Ross Sea. Micronuclei (MN) accounted for 10.50% of observed abnormalities in penguin erythrocytes while kidney-shaped nucleus (KSN) was the most abundant (20.88%). Heterophils (HE) were the most common WBC (36.93%) in agreement with the generic avian leukocytes profile while eosinophils (EO) were the lowest (7.45%). A low number of lymphocytes were detected resulting in a higher heterophils to lymphocytes ratio. ENAs and H:L ratio are confirmed as reliable indexes of penguin’s health status since they reflect their individual adaptation during breeding season. These baseline data will be useful for future studies as indicators of penguin’s health status mainly as response to environmental changes.

Nanoplastics are recognized as able to interact with other pollutants including heavy metals, and with natural organic matter, with implications for the potential risks to biota. We investigated the interaction of carboxylated polystyrene nanoparticles (PS–COOH NPs) with copper (Cu) and algal exudates (EPS) and how such interaction could affect Cu toxicity towards the freshwater microalga Raphidocelis subcapitata. PS–COOH NPs behavior in the presence of Cu and EPS was determined by dynamic light scattering (DLS), while PS–COOH NPs surface interaction with Cu ions and EPS was investigated by fluorimetric analysis. ICP-MS was used to test Cu ion adsorption to PS–COOH NPs in the presence and absence of algae. The interaction between PS–COOH NPs and the algal cell wall was assessed by fluorescence microscopy. Short- and long-term toxicity tests were carried out in parallel to assess the impact of PS–COOH NPs on algal growth. Results showed altered nanoparticle surface charge and hydrodynamic diameter following algal EPS exposure, supporting the hypothesis of a protein corona formation. In contrast, no absorption of Cu ions was observed on PS–COOH NPs, either in the presence or absence of algae. No differences on algal growth inhibition were observed between exposure to Cu only, and to Cu in combination with PS–COOH NPs, in short-term as well as long-term tests. However, after 72 h of exposure, the adsorption of PS-COOH NPs to algal cell walls appeared to correspond to morphological alterations, revealing potential disturbances in the mitotic cycle. Our findings confirm the ability of PS–COOH NPs to interact with EPS as shown for other nanomaterials. Environmentally realistic exposure scenarios are thus needed for evaluating nanoplastic toxicity, as nanoparticles will not maintain their pristine nature once released into natural media. Prolonged exposure and use of different end-points such as cell morphological changes and EPS production seem more reliable for the investigation of nanoplastic/algal cell interactions which can drive food chain transfer of nanoplastics and ultimately toxicity.

10 pages, 4 figures, Nowadays, the occurrence of a large volume of plastic litter in oceanic and coastal zones has increased concern about its impacts on marine organisms. The degradation of plastic polymers leads to the formation of smaller fragments at both micro and nano scale (, Marta Sendra is grateful to CEI·MAR for their support of the CEIJ-C06.1 project. This research has been funded by the Spanish National Research Plan MINECO (CTM2016-75908-R)

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.

The present study represents the first attempt to promote Ariadna spider as bioindicator of heavy elements contamination in the central region of the Namib Desert. Various spider populations belonging to undescribed Ariadna species have been described in Central Namib gravel plains and identified as sit-and-wait predators. By spending their life in individual tunnels, Ariadna spiders resemble the behaviour of ground-dwelling spiders that are well known as bioaccumulators of heavy elements. In the present study, 60 individuals of Ariadna spiders were collected from three populations located at various distances from major mining areas of Namib Desert. Contents of fifteen heavy elements were measured in sand samples taken around spider burrows as well as in spider bodies. Several oxidative stress responses as CAT and GST, MDA levels, aside cholinergic function (Cholinesterases), were assayed. Body burden of Ag, Cd, Cu and Zn resulted significantly higher than levels measured in sand, while Co, Cr, Ni, Pb, U and V levels were one or two order of magnitude lower in spider body than in sand samples. All enzymes activities (CAT, GST, ChE) as well as MDA levels were detectable in Ariadna spiders despite lower values than other species. Though we cannot affirm that all sampled spiders belong to the same species, their enzymatic activities reflect their ability to accumulate heavy elements regardless the specific habitat features, confirming so the value of this genus as bioindicator of heavy elements in the Central Namib Desert.

The uptake of Cd and some biomarkers of exposure and effects have been investigated in specimens of land snail Cornu aspersum exposed to vaporized CdCl2 (10 mg/L) for 7 days. The Cd levels quantified in snail's whole bodies confirmed Cd bioavailability trough vaporization and an higher accumulation in the midgut gland compared to the foot. Biological responses investigated showed a reduction of destabilization time of lysosomal membranes (NRRT) in hemocytes and an induction of catalase activities (CAT) in midgut gland. A further evidence of CdCl2 vaporized exposure was given by an increase in MT protein content as well as induction of Cd-MT gene expression, highlighting the central role of the midgut gland in Cd detoxification. These biomarkers can thus be considered as sensitive tools for the assessment of Cd contamination in the air using land snails as bioindicators. No changes in of GST activity and MDA were observed. From the overall results, the land snail, C. aspersum, could be used as good bioindicator of air quality for pollution monitoring purposes having shown clear signs of exposure and effects due Cd exposure by air.

The use of engineered nanomaterials (ENMs) for environmental remediation, known as nanoremediation, represents a challenging and innovative solution, ensuring a quick and efficient removal of pollutants from contaminated sites. Although the growing interest in nanotechnological solutions for pollution remediation, with significant economic investment worldwide, environmental and human risk assessment associated with the use of ENMs is still a matter of debate and nanoremediation is seen yet as an emerging technology. Innovative nanotechnologies applied to water and soil remediation suffer for a proper environmental impact scenario which is limiting the development of specific regulatory measures and the exploitation at European level. The present paper summarizes the findings from the workshop: “Ecofriendly Nanotechnology: state of the art, future perspectives and ecotoxicological evaluation of nanoremediation applied to contaminated sediments and soils” convened during the Biannual ECOtoxicology Meeting 2016 (BECOME) held in Livorno (Italy). Several topics have been discussed and, starting from current state of the art of nanoremediation, which represents a breakthrough in pollution control, the following recommendations have been proposed: (i) ecosafety has to be a priority feature of ENMs intended for nanoremediation; ii) predictive safety assessment of ENMs for environmental remediation is mandatory; (iii) greener, sustainable and innovative nano-structured materials should be further supported; (iii) those ENMs that meet the highest standards of environmental safety will support industrial competitiveness, innovation and sustainability. The workshop aims to favour environmental safety and industrial competitiveness by providing tools and modus operandi for the valorization of public and private investments.

Recent developments on environmental fate models indicate that as nano waste, engineered nanomaterials (ENMs) could reach terrestrial ecosystems thus potentially affecting environmental and human health. Plants can be therefore exposed to ENMs but controversial data in terms of fate and toxicity are currently available. Furthermore, there is a current lack of information on complex interactions/transformations to which ENMs undergo in the natural environment as for instance interacting with existing toxic compounds. The aim of the present study was to assess the behavior and biological effects of titanium dioxide nanoparticles (n-TiO2) (Aeroxide P25, Degussa Evonik) and its interaction with cadmium (CdCl2) in plants using radish seeds (Raphanus sativus L. Parvus) as model species. Radish seeds were exposed to n-TiO2 (1-1000mg/L) and CdCl2 (1-250mg/L) alone and in combination using a seed germination and seedling growth toxicity test OECD 208. Percentage of seed germination, germination index (GI) and root elongation were calculated. Cell morphology and oxidative stress parameters as glutathione-S-transferase (GST) and catalase activities (CAT) were measured in radish seeds after 5 days of exposure. Z-Average, PdI and Z-potential of n-TiO2 in Milli-Q water as exposure medium were also determined. DLS analysis showed small aggregates of n-TiO2, negative Z-potential and stable PdI in seed's exposure media. Germination percentage, GI and root length resulted affected by n-TiO2 exposure compared to controls. In particular, n-TiO2 at 1mg/L and 100mg/L did not affect radish seeds germination (100%) while at concentration of 10mg/L, 200mg/L, 500mg/L, and 1000mg/L a slight but not significant decrease of germination % was observed. Similarly root length and GI resulted significantly higher in seeds exposed to 10mg/L and 200mg/L compared to 1mg/L, 100mg/L, 500mg/L, 1000mg/L and control (p < 0.05). On the opposite, CdCl2 significantly abolished germination % and GI compared to control seeds and a concentration dependent decrease on root elongation was observed against controls (p < 0.05). As well, significant decrease of germination %, GI and root elongation was observed in seeds co-exposed to n-TiO2 and CdCl2 at the highest concentrations (1000mg/L n-TiO2 and 250mg/L CdCl2) compared to co-exposed seeds at low concentration (1mg/L n-TiO2 and 1mg/L CdCl2) and controls (p < 0.05). Root elongation significantly increase compared to control at the lowest co-exposure concentration (p < 0.05). Similarly at intermediate concentrations of 10 and 100mg/L in co-exposure conditions, n-TiO2 did not affect CdCl2 toxicity. Concerning antioxidant enzymes, a significant increase of CAT activity in seeds exposed to single high n-TiO2 concentration (1000mg/L) was observed while n-TiO2 (1mg/L), CdCl2 (1 and 250mg/L) and co-exposure resulted significantly decreased compared to controls (p < 0.05). Regarding GST activity, a slight increase in seeds exposed to 1000mg/L n-TiO2 but no significantly was observed, however both n-TiO2 and CdCl2 alone (1 and 250mg/L, respectively) or in combinations caused a significant decrease in GST activity (p < 0.05). Therefore, overall data support the hypothesis that the presence of n-TiO2 do not affect the toxicity of CdCl2 at least at the highest concentration (100 and 250mg/L) in radish seeds. Morphological alterations in nuclei, vacuoles and shape of radish root cells were observed upon single Cd exposure and not abolished in the presence of n-TiO2. Nevertheless, although n-TiO2 seems not to reduce Cd toxicity at high concentration (up to 250mg/L), interactions cannot be excluded based on obtained results.

The impact of nanoplastics using model polystyrene nanoparticles (PS NPs), anionic (PS-COOH) and cationic (PS-NH2), has been investigated on the marine rotifer Brachionus plicatilis, a major component of marine zooplanktonic species. The role of different surface charges in affecting PS NP behaviour and toxicity has been considered in high ionic strength media. To this aim, the selected media were standardized reconstituted seawater (RSW) and natural sea water (NSW), the latter resembling more natural exposure scenarios. Hatched rotifer larvae were exposed for 24h and 48h to both PS NPs in the range of 0.5-50μg/ml using PS NP suspensions made in RSW and NSW. No effects on lethality upon exposure to anionic NPs were observed despite a clear gut retention was evident in all exposed rotifers. On the contrary, cationic NPs caused lethality to rotifer larvae but LC50 values resulted lower in rotifers exposed in RSW (LC50=2.75±0.67µg/ml) compared to those exposed in NSW (LC50=6.62±0.87µg/ml). PS NPs showed similar pattern of aggregation in both high ionic strength media (RSW and NSW) but while anionic NPs resulted in large microscale aggregates (Z-average 1109 ± 128nm and 998±67nm respectively), cationic NP aggregates were still in nano-size forms (93.99 ± 11.22nm and 108.3 ± 12.79nm). Both PDI and Z-potential of PS NPs slightly differed in the two media suggesting a role of their different surface charges in affecting their behaviour and stability. Our findings confirm the role of surface charges in nanoplastic behaviour in salt water media and provide a first evidence of a different toxicity in rotifers using artificial media (RSW) compared to natural one (NSW). Such evidence poses the question on how to select the best medium in standardized ecotoxicity assays in order to properly assess their hazard to marine life in natural environmental scenarios.

Acid mine drainage (AMD) is a global environmental challenge that represents a primary source of contamination of heavy metals in both active and abandoned mining areas. The present study explored the suitability of a carboxymethylcellulose (CMC) hydrogel cross-linked with functionalized magnetite (Fe3O4) nanoparticles, (CMC-Fe3O4), for the removal of heavy metals from AMD-affected stream waters and tested its eco-friendly nature. AMD-affected stream waters, collected in a former mining site located in the Metalliferous Hills ore district (Central Italy), were chosen as a suitable benchmark for validating the CMC-Fe3O4 hydrogel applications. The remediation capability of the CMC-Fe3O4 hydrogel was investigated on solutions of Zn(II) in deionized (DI) water and on samples of AMD-affected stream waters. The CMC-Fe3O4 hydrogel was able to adsorb Zn(II) from DI water solutions as well as to reduce significantly the concentration of Zn and other heavy metals such as Cd, Co, Cu, Ni and Pb, in AMD-affected stream waters. CMC-Fe3O4 hydrogel and treated waters (both Zn-solution and AMD-affected stream waters) were tested for their effects on the freshwater green alga R. subcapitata by using 72-h growth inhibition tests. No effects were observed on the R. subcapitata growth upon exposure to CMC-Fe3O4 hydrogel, suggesting its eco-friendly nature. Moreover, CMC-Fe3O4 hydrogel was able to reduce the toxicity of heavy metals to the green alga, both in DI and AMD-affected stream water. Based on our findings, CMC-Fe3O4 hydrogel could be considered as a valuable tool for an environmentally safe treatment of AMD-affected stream waters.

Polystyrene nanoparticles have been shown to pose serious risk to marine organisms including sea urchin embryos based on their surface properties and consequently behaviour in natural sea water. The aim of this study is to investigate the toxicity pathways of amino polystyrene nanoparticles (PS-NH2, 50 nm) in Paracentrotus lividus embryos in terms of development and signalling at both protein and gene levels. Two sub-lethal concentrations of 3 and 4 μg/mL of PS-NH2 were used to expose sea urchin embryos in natural sea water (PS-NH2 as aggregates of 143 ± 5 nm). At 24 and 48 h post-fertilisation (hpf) embryonic development was monitored and variations in the levels of key proteins involved in stress response and development (Hsp70, Hsp60, MnSOD, Phospho-p38 Mapk) as well as the modulation of target genes (Pl-Hsp70, Pl-Hsp60, Pl-Cytochrome b, Pl-p38 Mapk, Pl-Caspase 8, Pl-Univin) were measured. At 48 hpf various striking teratogenic effects were observed such as the occurrence of cells/masses random...

Nanoremediation, which is the use of nanoparticles and nanomaterials for environmental remediation, is widely explored and proposed for preservation of ecosystems that suffer from the increase in human population, pollution, and urbanization. We herein report a critical analysis of nanotechnologies for water remediation by assessing their sustainability in terms of efficient removal of pollutants, appropriate methods for monitoring their effectiveness, and protocols for the evaluation of any potential environmental risks. Our purpose is to furnish fruitful guidelines for sustainable water management, able to promote nanoremediation also at European level. In this context, we describe new nanostructured polysaccharide-based materials obtained from renewable resources as alternative efficient and ecosafe solutions for water nano-treatment. We also provide eco-design indications to improve the sustainability of the production of these materials, based on life-cycle assessment methodology.

Aim of this study was to evaluate the responsiveness of red mullet (Mullus barbatus) liver detoxification enzymes to PAHs at transcriptional and post-transcriptional levels in the field. Fish were captured in the north-eastern Adriatic Sea, close to an oil refinery. Sixteen PAHs (EPA) were determined in sediments and fish fillets; transcription levels of cyp1a, cyp3a and abcc2 genes and EROD, BROD, B(a)PMO, BFCOD, GST and UDPGT enzymatic activities were measured. Levels of PAHs in sediments reflect the oil pollution gradient of the area, with weak correspondence in fish fillets. cyp1a gene transcription and EROD, B(a)PMO and BFCOD activities were significantly induced in the oil refinery site, and a slight up-regulation of cyp3a and abcc2 was also observed. GST and UDPGT remained unchanged. The present study provides the first data on detoxification responses at transcriptional levels in the liver of red mullet and confirms phase I enzymes as suitable biomarkers of exposure to PAHs in field studies.

Despite the growing concern over the potential biological impact of nanoparticles (NPs) in the aquatic environment, little is known about their interactions with other pollutants. In the marine mussel Mytilus galloprovincialis, exposure to nanosized titanium dioxide (n-TiO2), one of the most widespread type of NPs in use, in combination with and 2,3,7,8-tetrachlorodibenzo-p-dioxins (TCDD), chosen as model organic xenobiotic, was shown to induce significant changes in different biomarkers in hemocytes, gills and digestive gland, with distinct effects depending on cell/tissue and type of response measured. In this work, the interactive effects of n-TiO2 and TCDD at the tissue level were further investigated in mussel digestive gland using an integrated approach transcriptomics/immunohistochemistry. Mussels were exposed to n-TiO2 (100μgL(-1)) and TCDD (0.25μgL(-1)), alone and in combination, for 96h. Transcriptomic analysis identified 48-, 49- and 62 Differentially Expressed Genes (DEGs) in response to n-TiO2, TCDD and n-TiO2/TCDD, respectively. Gene Ontology (GO) term analysis revealed distinct biological processes affected in different experimental conditions. n-TiO2 mainly up-regulated cytoskeletal genes, while TCDD up-regulated endocrine and signal transduction related processes. Co-exposure induced transcriptional changes common to individual treatments, and identified a newly generated process, response to chemical stimulus. Transcription of selected genes was verified by qPCR. Moreover, expression of tubulin, as an example of target protein of interest identified by gene transcription data, was evaluated in tissue sections by immunolabelling. Tissue TCDD accumulation was evaluated by immunofluorescence with an anti-dioxins antibody. The results demonstrate both distinct and interactive effects of n-TiO2 and TCDD in mussel digestive gland at the molecular and tissue level, identify the main molecular targets involved, and underline how exposure to the n-TiO2/TCDD mixture does not result in increased TCDD accumulation and overall stressful conditions in the tissue. These represent the first data on transcriptional responses of marine invertebrates to exposure not only to n-TiO2 as a model of NP, but also to a legacy contaminant like TCDD.

The development of nanotechnology will inevitably lead to the release of consistent amounts of nanomaterials (NMs) and nanoparticles (NPs) into marine ecosystems. Ecotoxicological studies have been carried out to identify potential biological targets of NPs, and suitable models for predicting their impact on the health of the marine environment. Recent studies in invertebrates mainly focused on NP accumulation and sub-lethal effects, rather than acute toxicity. Among marine invertebrates, bivalves represent by large the most studied group, with polychaetes and echinoderms also emerging as significant targets of NPs. However, major scientific gaps still need to be filled. In this work, factors affecting the fate of NPs in the marine environment, and their consequent uptake/accumulation/toxicity in marine invertebrates will be summarized. The results show that in different model species, NP accumulation mainly occurs in digestive tract and gills. Data on sub-lethal effects and modes of action of different types of NPs (mainly metal oxides and metal based NPs) in marine invertebrates will be reviewed, in particular on immune function, oxidative stress and embryo development. Moreover, the possibility that such effects may be influenced by NP interactions with biomolecules in both external and internal environment will be introduced. In natural environmental media, NP interactions with polysaccharides, proteins and colloids may affect their agglomeration/aggregation and consequent bioavailability. Moreover, once within the organism, NPs are known to interact with plasma proteins, forming a protein corona that can affect particle uptake and toxicity in target cells in a physiological environment. These interactions, leading to the formation of eco-bio-coronas, may be crucial in determining particle behavior and effects also in marine biota. In order to classify NPs into groups and predict the implications of their release into the marine environment, information on their intrinsic properties is clearly insufficient, and a deeper understanding of NP eco/bio-interactions is required.

The bivalve Mytilus galloprovincialis has proven as a suitable model invertebrate for evaluating the potential impact of nanoparticles (NPs) in the marine environment. In particular, in mussels, the immune system represents a sensitive target for different types of NPs. In environmental conditions, both NP intrinsic properties and those of the receiving medium will affect particle behavior and consequent bioavailability/uptake/toxicity. However, the evaluation of the biological effects of NPs requires additional understanding of how, once within the organism, NPs interact at the molecular level with cells in a physiological environment. In mammalian systems, different NPs associate with serum soluble components, organized into a "protein corona", which affects particle interactions with target cells. However, no information is available so far on the interactions of NPs with biological fluids of aquatic organisms. In this work, the influence of hemolymph serum (HS) on the in vitro effects of amino modified polystyrene NPs (PS-NH2) on Mytilus hemocytes was investigated. Hemocytes were incubated with PS-NH2 suspensions in HS (1, 5 and 50µg/mL) and the results were compared with those obtained in ASW medium. Cell functional parameters (lysosomal membrane stability, oxyradical production, phagocytosis) were evaluated, and morphological changes were investigated by TEM. The activation state of the signalling components involved in Mytilus immune response (p38 MAPK and PKC) was determined. The results show that in the presence of HS, PS-NH2 increased cellular damage and ROS production with respect to ASW medium. The effects were apparently mediated by disregulation of p38 MAPK signalling. The formation of a PS-NH2-protein corona in HS was investigated by centrifugation, and 1D- gel electrophoresis and nano-HPLC-ESI-MS/MS. The results identified the Putative C1q domain containing protein (MgC1q6) as the only component of the PS-NH2 hard protein corona in Mytilus hemolymph. These data represent the first evidence for the formation of a NP bio-corona in aquatic organisms and underline the importance of the recognizable biological identity of NPs in physiological exposure medium when testing their potential impact environmental model organisms. Although the results obtained in vitro do not entirely reflect a realistic exposure scenario and the more complex formation of a bio-corona that is likely to occur in vivo, these data will contribute to a better understanding of the effects of NPs in marine invertebrates.

Nano-sized polymers as polystyrene (PS) constitute one of the main challenges for marine ecosystems, since they can distribute along the whole water column affecting planktonic species and consequently disrupting the energy flow of marine ecosystems. Nowadays very little knowledge is available on the impact of nano-sized plastics on marine organisms. Therefore, the present study aims to evaluate the effects of 40 nm anionic carboxylated (PS-COOH) and 50 nm cationic amino (PS-NH2) polystyrene nanoparticles (PS NPs) on brine shrimp Anemia franciscana larvae. No signs of mortality were observed at 48 h of exposure for both PS NPs at naplius stage but several sub-lethal effects were evident. PS-COOH (5-100 mu g/ml) resulted massively sequestered inside the gut lumen of larvae (48 h) probably limiting food intake. Some of them were lately excreted as fecal pellets but not a full release was observed. Likewise, PS-NH2 (5-100 mu g/ml) accumulated in larvae (48 h) but also adsorbed at the surface of sensorial antennules and appendages probably hampering larvae motility. In addition, larvae exposed to PS-NH2 undergo multiple molting events during 48 h of exposure compared to controls. The activation of a defense mechanism based on a physiological process able to release toxic cationic NPs (PS-NH2) from the body can be hypothesized. The general observed accumulation of PS NPs within the gut during the 48 h of exposure indicates a continuous bioavailability of nano-sized PS for planktonic species as well as a potential transfer along the trophic web. Therefore, nano-sized PS might be able to impair food uptake (feeding), behavior (motility) and physiology (multiple molting) of brine shrimp larvae with consequences not only at organism and population level but on the overall ecosystem based on the key role of zooplankton on marine food webs. (C) 2015 Elsevier Inc. All rights reserved.

Biological responses and contaminant levels in biological tissues were investigated in fish specimens of grass goby (Zosterisessor ophiocephalus) collected in spring 2000 in a moderately polluted brackish ecosystem, the Orbetello lagoon (southern coast of Tuscany, Italy). The complexity of harmful effects that contaminants may cause in fish over time was assessed. General condition indices (somatic liver index, and gonadal somatic index), cytochrome P450 enzymes, acetylcholinesterase activity and ovarian morphology were integrated with the content of organochlorines (OCs-hexachlorobenzene, DDTs and polychlorinated biphenyls) in liver and ovaries and p-nonylphenol (NP) and lower ethoxylate (NPE(1-2)) content in muscle and ovaries as well. Samples collected from the western basin of the lagoon near the Orbetello sewage treatment plant had somatic liver index values and two P450 activities significantly higher than those collected from the same basin at the Albegna river delta and in the Ansedonia canal in the eastern basin (P < 0.05). On the other hand, while muscle content of p-NP and NPE(1-2) was significantly higher in samples from the Orbetello and Albegna river sites than in samples from the Ansedonia canal site, OC levels in liver (DDTs > PCBs > HCB) were particularly lower in samples from STP compared to levels detected in sites AC and AR. Light microscopy of ovaries revealed that although the gonads were sexually mature, the diameter of the oocytes and eggs was smaller than that commonly reported. High levels of M and NPs were, in fact, observed in grass goby ovaries, and the differences observed between acetylcholinesterase activities among investigated sites suggest exposure to insecticides as well. The integration of biological responses and contaminant tissue content reveals that certain areas of the lagoon are selectively impacted by various human activities and helps in distinguishing relationships between biological responses and contaminant load in the lagoon environment., Les réponses biologiques et les niveaux de contamination des tissus ont été recherchés dans des spécimens de gobies d'herbiers (Zosterisessor ophiocephalus) récoltés au printemps 2000 dans un écosystème saumâtre modérément pollué, la lagune d'Orbetello (côte sud de la Toscane). La complexité des effets nuisibles des contaminants sur les poissons est suivie au cours du temps. Des indices globaux (indice somatique hépatique, indice somatique gonadique), les enzymes du cytochrome P450, l'activité acétylcholinestérase et la morphologie de l'ovaire sont utilisés ainsi que le contenu organochloré (hexachlorobenzène, DDTs et biphényles polychlorinatés) du foie et des ovaires, les teneurs en p-nonylphénol et en éthoxylates inférieurs du muscle et des ovaires. Des échantillons provenant du bassin ouest de la lagune près de la station d'épuration d'Orbetello présentent, pour l'indice somatique hépatique et pour les deux activités P450, des valeurs nettement supérieures à celles qui sont relevées dans le même bassin dans le delta de la rivière Albegna et dans le canal Ansedonia dans le bassin est (P < 0,05). D'autre part, alors que le contenu des muscles en p-nonylphénol et en éthoxylates inférieurs apparaît nettement supérieur dans les échantillons provenant des sites « station d'épuration d'Orbetello » et « rivière Albegna » comparé aux échantillons du site « canal Ansedonia », les niveaux d'organochlorés dans le foie (DDTs > PCBs > HCB) sont particulièrement faibles au site « station d'épuration d'Orbetello » comparé à ceux des sites « canal Ansedonia » et « rivière Albegna ». L'examen microscopique des ovaires révèle que, même si les gonades sont sexuellement mûres, le diamètre des oocytes et des œufs est plus petit qu'habituellement. De hautes concentrations d'hexachlorobenzènes et de p-nonylphenols sont en fait observées dans les ovaires des gobies des herbiers et les différences entre les activités acetylcholinestérase des différents sites suggèrent une exposition aux insecticides. L'intégration des réponses biologiques et du contenu tissulaire en contaminants indique que certaines aires de la lagune subissent un impact sélectif des activités humaines. Ceci met en évidence les relations entre les réponses biologiques et la charge en contaminants dans l'environnement lagunaire.