Lowering the pH leads to the disaggregation of NiO and ZnO nanoparticles and modifies the mutagenic response.

In a changing environmental scenario, acid rain can have a significant impact on aquatic ecosystems. Acidification is known to produce corrosion in metals, hence increasing their harmful effects on the environment, organisms and human health. The prevalent use of metallic nanoparticles (NPs) in everyday products raises concerns regarding exposure and nanotoxicity even in these acidified conditions. We thus report on the cytotoxic and genotoxic potential of nickel oxide (NiO-NP) and zinc oxide (ZnO-NP) NPs when suspended in aqueous media in light of pH variations (7.5 and 5). A modified microsuspension method of the Salmonella/microsome assay was adopted, and strains (TA97a, TA98, TA100, TA102) were exposed to NPs (10-1280 µg/plate) with and without a metabolization fraction. The acidic condition favored disaggregation and caused a decrease in NPs size. Mutagenicity was observed in all samples and different strains, with greater DNA base pair substitution damage (TA100 and TA102), but extrinsic conditions (pH) suggest different action mechanisms of NiO-NP and ZnO-NP on genetic content. Mutagenic activity was found to increase upon metabolic activation (TA98, TA100, and TA102) demonstrating the bioactivity of NiO-NP and ZnO-NP in relation to metabolites generated by the mammalian p450 system in vitro. Modifications in the Salmonella assay methodology increased cell exposure time. The observed responses recommend this modified assay as one of the methodologies of choice for nanoecotoxicological evaluation. These findings emphasize the significance of incorporating the environmental context when evaluating the toxicity of metal-based NPs.

The Effect of Silver Nanoparticles on the Digestive System, Gonad Morphology, and Physiology of Butterfly Splitfin (Ameca splendens).

The aim of this study was to determine the effects of silver nanoparticles (AgNPs) on the morphology and enzymatic activity of butterfly splitfin (Ameca splendens). Individuals of both sexes, aged about five months, were exposed to AgNPs at concentrations of 0 (control group), 0.01, 0.1, and 1.0 mg/dm3 for 42 days. On the last day of the experiment, the fish were euthanized, subjected to standard histological processing (anterior intestine, liver, and gonads), and analysed for digestive enzyme activity in the anterior intestine and oxidative stress markers in the liver. Fish in the AgNP 0.01 and 0.1 groups had the lowest anterior intestinal fold and enterocyte height. However, there were no statistically significant changes in the digestive enzyme activity in the anterior intestine. Analysis of enzymatic activity in the liver showed an increase in superoxide dismutase activity in fish in the AgNP 0.1 group. Histological analyses showed that AgNPs inhibited meiotic divisions at prophase I in a non-linear manner in ovaries and testes. In the AgNP 0.1 and 1.0 groups, the area occupied by spermatocytes was lower compared to the other groups. These results indicate that exposure to AgNPs may lead to disturbances in morphology and enzymatic activity in the liver and intestine and may lead to disruption of reproduction in populations.

Graphene oxide-silver nanoparticle hybrid material: an integrated nanosafety study in zebrafish embryos.

This work reports an integrated nanosafety study including the synthesis and characterization of the graphene oxide-silver nanoparticle hybrid material (GO-AgNPs) and its nano-ecotoxicity evaluation in the zebrafish embryo model. The influences of natural organic matter (NOM) and a chorion embryo membrane were considered in this study, looking towards more environmentally realistic scenarios and standardized nanotoxicity testing. The nanohybrid was successfully synthesized using the NaBH4 aqueous method, and AgNPs (~ 5.8 nm) were evenly distributed over the GO surface. GO-AgNPs showed a dose-response acute toxicity: the LC50 was 1.5 mg L-1 for chorionated embryos. The removal of chorion, however, increased this toxic effect by 50%. Furthermore, the presence of NOM mitigated mortality, and LC50 for GO-AgNPs changed respectively from 2.3 to 1.2 mg L-1 for chorionated and de-chorionated embryos. Raman spectroscopy confirmed the ingestion of GO by embryos; but without displaying acute toxicity up to 100 mg L-1, indicating that the silver drove toxicity down. Additionally, it was observed that silver nanoparticle dissolution has a minimal effect on these observed toxicity results. Finally, understanding the influence of chorion membranes and NOM is a critical step towards the standardization of testing for zebrafish embryo toxicity in safety assessments and regulatory issues.

Effects of multiwalled carbon nanotubes co-exposure with cadmium on zebrafish cell line: Metal uptake and accumulation, oxidative stress, genotoxicity and cell cycle.

Carbon nanotubes presence in the environment increases every year because of exponential industrial production around the world. In aquatic environments, carbon nanotubes can interact with other pollutants based on their adsorbent surface chemistry properties. Heavy metal ions represent one of the biggest concerns in water resources nowadays due to anthropogenic activities, in which cadmium (Cd) is one of the most harmful metal for aquatic organisms. This study investigated the influence of two co-exposure protocols differing by the order of interaction of oxidized multiwalled carbon nanotubes (ox-MWCNT) with Cd in zebrafish liver cell line (ZFL). The ox-MWCNT was characterized, Cd content in culture medium and uptake by cells were quantified using ICP-MS and, the reactive oxygen species (ROS), the biotransformation enzymes activity of phase I and II as well as the antioxidants defenses and oxidative damage were analyzed. The effects on the cell cycle were investigated by flow cytometry and DNA damage by comet assay. The exposure to ox-MWCNT alone decreased the activity of catalase, glutathione peroxidase, and glutathione S-transferase and altered the cell cycle with a reduction of cells in the G2/M phase. Cd exposure alone decreased the activity of catalase and glutathione S-transferase, increased ROS, metallothionein, and lipid peroxidation content and causes genotoxicity in the cells. Despite different incubation protocol, the co-exposure ox-MWCNT-Cd increased the Cd content in ZFL cells after 24 h exposure, increased ROS production and DNA damage without differences between them. Our results showed the modulation of ox-MWCNT on Cd effects and contributed to future co-exposure toxicity investigations and nanosafety regulations involving carbon nanomaterials and aquatic pollutants.

Influence of wastewater type on the impact generated by TiO2 nanoparticles on the oxygen uptake rate in activated sludge process.

Physicochemical characteristics of wastewater have a relationship with the stability of TiO2 nanoparticles (NPs). This in turn has an effect on the toxicity of TiO2 NPs in microorganisms. In this work, the effect of TiO2 NPs on activated sludge process was evaluated using three different types of wastewater: synthetic, raw, and filtered. The results showed that aggregate size of TiO2 NPs and their specific adsorption of substrates were influenced by the type of substrates and the presence of suspended solids in the wastewater. It was also shown that TiO2 NPs in raw wastewater severely inhibited oxygen uptake by microorganisms as compared to uptake in synthetic or filtered wastewater. The attachment of TiO2 NP aggregates on cell membranes was observed for all types of wastewater. However, the internalization of TiO2 NPs by microorganisms was observed only for raw and filtered wastewater. These results indicate that the effects caused by TiO2 NPs on activated sludge were different depending on the wastewater used for the experiment.

Influence of pH and media composition on suspension stability of silver, zinc oxide, and titanium dioxide nanoparticles and immobilization of Daphnia magna under guideline testing conditions.

In aquatic toxicity testing of engineered nanoparticles (ENPs) the process of agglomeration is very important as it may alter bioavailability and toxicity. In the present study, we aimed to identify test conditions that are favorable for maintaining stable ENP suspensions. We evaluated the influence of key environmental parameters: pH (2-12) and ionic strength using M7, Soft EPA (S EPA) medium, and Very Soft EPA (VS EPA) medium; and observed the influence of these parameters on zeta potential, zeta average, and acute immobilization of Daphnia magna for three different ENPs. Despite being sterically stabilized, test suspensions of silver (Ag) ENPs formed large agglomerates in both VS EPA and M7 media; and toxicity was found to be higher in VS EPA medium due to increased dissolution. Low-agglomerate suspensions for zinc oxide (ZnO) could be obtained at pH 7 in VS EPA medium, but the increase in dissolution caused higher toxicity than in M7 medium. Titanium dioxide (TiO2) ENPs had a point of zero charge in the range of pH 7-8. At pH 7 in VS EPA, agglomerates with smaller hydrodynamic diameters (~200nm) were present compared to the high ionic strength M7 medium where hydrodynamic diameters reached micrometer range. The stable suspensions of TiO2 ENPs caused immobilization of D. magna, 48-h EC50 value of 13.7mgL(-1) (95% CI, 2.4mg-79.1mgL(-1)); whereas no toxicity was seen in the unstable, highly agglomerated M7 medium suspensions, 48-h EC50 >100mgL(-1). The current study provides a preliminary approach for methodology in testing and assessing stability and toxicity of ENPs in aquatic toxicity tests of regulatory relevance.

Phytotoxicity of ionic, micro- and nano-sized iron in three plant species.

Potential environmental impacts of engineered nanoparticles (ENPs) can be understood taking into consideration phytotoxicity. We reported on the effects of ionic (FeCl3), micro- and nano-sized zerovalent iron (nZVI) about the development of three macrophytes: Lepidium sativum, Sinapis alba and Sorghum saccharatum. Four toxicity indicators (seed germination, seedling elongation, germination index and biomass) were assessed following exposure to each iron concentration interval: 1.29-1570mg/L (FeCl3), 1.71-10.78mg/L (micro-sized iron) and 4.81-33,560mg/L (nano-iron). Exposure effects were also observed by optical and transmission electron microscopy. Results showed that no significant phytotoxicity effects could be detected for both micro- and nano-sized zerovalent irons, including field nanoremediation concentrations. Biostimulation effects such as an increased seedling length and biomass production were detected at the highest exposure concentrations. Ionic iron showed slight toxicity effects only at 1570mg/L and, therefore, no median effect concentrations were determined. By microscopy, ENPs were not found in palisade cells or xylem. Apparently, aggregates of nZVI were found inside S. alba and S. saccharatum, although false positives during sample preparation cannot be excluded. Macroscopically, black spots and coatings were detected on roots of all species especially at the most concentrated treatments.

Direct and indirect toxic effects of cotton-derived cellulose nanofibres on filamentous green algae.

Recently, cellulose nanofibers (CNFs) have attracted considerable attention as natural, abundant polymers with excellent mechanical properties and biodegradability. CNFs provide a new materials platform for the sustainable production of high-performance nano-enable products for various applications. Given the increasing rates of CNF production, the potential for their release to the environment and the subsequent impact on ecosystem is becoming an increasing concern that needs to be addressed. Here, we used the Klebsormidium flaccidum as a bioindicator organism of terrestrial and freshwater habitats pollution using a battery of biomarkers. Our results show that cotton CNFs inhibit the proliferation of algae and induce morphological changes in them. The two main toxicity mechanisms induced by cotton CNFs are: (i) a direct contact of CNFs with the cell wall and cellular membrane and (ii) an indirect effect through the generation of reactive oxygen species (ROS).

Physiological changes of the lichen Parmotrema tinctorum as result of carbon nanotubes exposition.

Carbon nanotubes (CNT) is one of the more abundant nanomaterial produced in the world. Therefore, it is desirable to access its effects in all environment compartments, in order to mitigate environmental distress. This study aims to verify the potential use of lichens - classical atmospheric pollution indicators - as biomonitors of carbon nanotubes aerosols. To examine cause-effect relationships, preserving environmental microclimatic parameters, the lichen Parmotrema tinctorum (Nyl.) Hale was transplanted to open top chambers where aerosols of CNT were daily added. Physiological parameters such as cell viability, photosynthetic efficiency, cell permeability as well as nanoparticle internalization were assessed. Carbon nanotubes exposure led to reduction on the cell viability of P. tinctorum. The treatment with 100µg/mL of MWCNT-COOH resulted in intracellular ion leakage, probably due to changes in membrane permeability. No alterations on photosynthetic efficiency were detected. Carbon nanotubes entrapment and internalization into the lichen thallus were observed. Short term exposition of CNT produced measurable physiological changes in P. tinctorum lichen. This suggests the possibility of use of lichens as models to assess the environmental impact (air related) of engineered nanomaterials.

Co-exposure of iron oxide nanoparticles with glyphosate herbicides in Poecilia reticulata: Fish liver damages is reversible during iron accumulation and elimination period.

Iron oxide nanoparticles (IONPs) are advanced materials for water remediation technologies. It is therefore relevant to evaluate the cellular and tissue behavior of fishes in response to IONPs and their associations with agrochemicals such as glyphosate (GLY) and glyphosate-based herbicides (GBHs). Iron accumulation, tissue integrity and lipid distribution in the hepatocytes of Poecilia reticulata (guppy) were investigated in a control group and in groups exposed to soluble iron ions, namely IFe (0.3 mgFe/L), IONPs (0.3 mgFe/L), and IONPs, associated with GLY (0.65 mg/L), GBHs 0.65 mgGLY/L (IONPs + GBH1), and 1.30 mgGLY/L (IONPs + GBH2), for 7, 14, and 21 days, followed by an equal period of postexposure in clean reconstituted water. The results showed that the accumulation of iron was greater in the subjects in the IONP treatment group when compared to that in the Ife group. In addition, the subjects in the mixtures with GBHs had a greater accumulation of iron than those in the IONP + GLY treatment group. Tissue integrity assessments demonstrated an intense accumulation of lipids, formation of necrotic zones and leukocyte infiltrates in all the treated groups, with a greater quantity of lipids in the animals treated with IONP + GLY and IFe. During postexposure, the results indicated an elimination of iron in all treated groups, reaching the same level as the control group, throughout the 21 days postexposure. Thus, the damage caused to animal livers by IONP mixtures is reversible, providing promising results for the development of safe environmental remediation practices using nanoparticles.

Uptake and Effects of Nanoplastics on the Dinoflagellate Gymnodinium corollarium.

Plastic nanoparticles (NPs) are the final state of plastic degradation in the environment before they disintegrate into low-molecular-weight organic compounds. Unicellular organisms are highly sensitive to the toxic effects of nanoplastics, because they are often capable of phagotrophy but are unable to consume a foreign material such as synthetic plastic. We studied the effect of polystyrene, poly(vinyl chloride), poly(methyl acrylate), and poly(methyl methacrylate) NPs on the photosynthetic dinoflagellate Gymnodinium corollarium Sundström, Kremp et Daugbjerg. Fluorescent tagged particles were used to visualize plastic capture by dinoflagellate cells. We found that these dinoflagellates are capable of phagotrophic nutrition and thus should be regarded as mixotrophic species. This causes their susceptibility to the toxic effects of plastic NPs. Living cells ingest plastic NPs and accumulate in the cytoplasm as micrometer-level aggregates, probably in food vacuoles. The action of nanoplastics leads to a dose-dependent increase in the level of reactive oxygen species in dinoflagellate cells, indicating plastic degradation in the cells. The introduction of a methyl group into the main chain in the α-position in the case of poly(methyl methacrylate) causes a drastic reduction in toxicity. We expect that such NPs can be a tool for testing unicellular organisms in terms of heterotrophic feeding ability. We suggest a dual role of dinoflagellates in the ecological fate of plastic waste: the involvement of nanoplastics in the food chain and its biochemical destruction. Environ Toxicol Chem 2023;42:1124-1133. © 2023 SETAC.

"Smart" nanosensors for early detection of corrosion: Environmental behavior and effects on marine organisms.

Corrosion is an environmental and economic global problem. "Smart" or stimuli-responsive colorimetric nanosensors for maritime coatings have been proposed as an asset to overcome the limitations of the current monitoring techniques by changing color in the presence of triggers associated with the early stages of corrosion. Layered double hydroxides (Zn-Al LDH; Mg-Al LDH) and silica mesoporous nanocapsules (SiNC) were used as precursor nanocarriers of active compounds: hexacyanoferrate ions ([Fe(CN)6]3-) and phenolphthalein (PhPh), respectively. Additionally, the safer-by-design principles were employed to optimize the nanosensors in an eco-friendly perspective (e.g., regular vs. warm-washed SiNC-PhPh; immobilization using different carriers: Zn-Al LDH-[Fe(CN)6]3- vs. Mg-Al LDH-[Fe(CN)6]3-). Therefore, the present study aims to assess the environmental behavior in saltwater and the toxic effects of the nanosensors, their nanocarriers, and the active compounds on the marine microalgae Tetraselmis chuii and the crustacean Artemia salina. Briefly, tested compounds exhibited no acute toxic effects towards A. salina (NOEC = 100 mg/L), apart from SiNC-PhPh (LC50 = 2.96 mg/L) while tested active compounds and nanosensors caused significant growth inhibition on T. chuii (lowest IC50 = 0.40 mg/L for SiNC-PhPh). The effects of [Fe(CN)6]3- were similar regardless of the nanocarrier choice. Regarding SiNC-PhPh, its toxicity can be decreased at least twice by simply reinforcing the nanocapsules washing, which contributes to the removal (at least partially) of the surfactants residues. Thus, implementing safe-by-design strategies in the early stages of research proved to be critical, although further progress is still needed towards the development of truly eco-friendly nanosensors.

Exposure of Midge Larvae (Chironomus riparius) to Graphene Oxide Leads to Development Alterations.

Despite the fast-growing use and production of graphene-based nanomaterials (GBMs), data concerning their effects on freshwater benthic macroinvertebrates are scarce. This study aims to investigate the effects of graphene oxide (GO) on the midge Chironomus riparius. Mortality, growth inhibition, development delay and teratogenicity, assessed using mentum deformity analysis, were investigated after a 7-day static exposure of the first instar larvae under controlled conditions. The collected data indicated that the survival rate was not impacted by GO, whereas chronic toxicity following a dose-dependent response occurred. Larval growth was affected, leading to a significant reduction in larval length (from 4.4 to 10.1%) in individuals reaching the fourth instar at any of the tested concentrations (from 0.1 to 100 mg/L). However, exposure to GO is not associated with an increased occurrence of mouthpart deformities or seriousness in larvae. These results highlight the suitability of monitoring the larval development of C. riparius as a sensitive marker of GO toxicity. The potential ecological consequences of larval size decrease need to be considered for a complete characterization of the GO-related environmental risk.

Bioaccumulation and depuration of TiO2 nanoparticles by zebrafish through dietary exposure: Size- and number concentration-resolved analysis using single-particle ICP-MS.

The bioaccumulation and depuration of TiO2 nanoparticles (TiO2NPs) by zebrafish via the dietary exposure following the OECD Test Guideline 305 (OECD TG305) was evaluated using particle size- and number concentration-resolved analysis based on single-particle ICP-MS (spICP-MS). We found that using enzymatic digestion without H2O2 or excessive heating can recover 84.0 ± 4.0% and 94.5 ± 3.5% of TiO2NP mass and number concentrations from fish tissue, respectively, without altering the size distribution of parent TiO2NPs. OECD TG305 can allow for the evaluation of bioaccumulation and depuration of TiO2NPs by fish based on the particle mass and number dose metrics. The toxicokinetic modeling can reasonably describe the mass- and number-based measurement data with the derived absorption efficiency α at ~0.2, depuration rate at ~0.5 d-1, and kinetic biomagnification factor (BMFk) at ~0.007 comparable with available data. The mass concentration- and number concentration-based bioaccumulation metrics including body burdens are correlated for TiO2NPs that remained nano-sized in vivo and exhibited marginal physicochemical alterations upon uptake by fish. The result indicates that the traditional mass concentration metric may be used to represent the fish bioaccumulation potential for chemically inert NPs like TiO2.

Impacts of engineered iron nanoparticles on oxidative stress, fatty acid composition, and histo-architecture of the smooth scallop Flexopecten glaber.

Engineered iron nanoparticles are widely used in environmental remediation, yet their potential toxic effects on marine biota remain poorly elucidated. This study aimed to gain insight into the nanoscale zero-valent iron (NZVI) toxicity mechanisms for marine invertebrates. Aside from the effect on oxidative status and histopathology, the effect of NZVI on lipid metabolism in bivalves was studied for the first time. To this end, specimens of Flexopecten glaber were exposed to ascending concentrations (0.5, 1, and 1.5 mg/L) of NZVI for 96 h. Results illustrate differential patterns of iron accumulation in the gills and the digestive gland. By increasing NZVI concentrations, the total iron level tended to markedly increase in the gills and decrease in the digestive gland, reaching 132 and 37.6 µg/g DW, respectively, in the specimens exposed to 1.5 mg/L. Biochemical and cellular biomarkers highlighted that NZVI caused oxidative stress (measured as hydrogen peroxide, malondialdehyde, and advanced oxidation protein product levels) and alterations of antioxidant defense systems, including reduced glutathione, non-protein thiol, glutathione peroxidase, superoxide dismutase, and catalase. Modulation of lipid metabolism with changed fatty acid compositions (mainly an increase in the saturation and a decrease in unsaturation levels) was also observed in both gills and digestive gland. Moreover, several histological damages, including lipofuscin accumulation, infiltrative inflammations, and digestive tubule alterations, were observed in the two studied organs, providing supplementary evidence regarding the toxic effect of NZVI. This study adds to the growing body of evidence pointing to the hazardous impacts of iron NPs on aquatic ecosystems.

Form-Specific and Probabilistic Environmental Risk Assessment of 3 Engineered Nanomaterials (Nano-Ag, Nano-TiO2 , and Nano-ZnO) in European Freshwaters.

The release of engineered nanomaterials (ENMs) to the environment necessitates an assessment of their environmental risks. The currently available environmental risk assessments (ERA) for ENMs are based on an analysis of the total flows of a specific ENM to the environment and on ecotoxicity studies performed with pristine ENMs. It is known that ENMs undergo transformation during product use and release and in technical systems such as wastewater treatment. The aim of the present study was therefore to perform an ERA of 3 ENMs (nano-Ag, nano-TiO2 , and nano-ZnO) based on a form-specific release model and a form-specific analysis of ecotoxicological data. Predicted environmental concentration values were derived using a form-specific material flow model. Species sensitivity distributions were used to derive predicted-no-effect concentrations (PNECs) for the pristine ENMs and for dissolved and transformed Ag and ZnO. For all ENMs, the matrix-embedded form was included in the assessment. A probabilistic assessment was applied, yielding final probability distributions for the risk characterization ratio (RCR). For nano-Ag, the form-specific assessment resulted in a decrease of the mean RCR from 0.061 for the approach neglecting the different release forms to 0.034 because of the much lower PNEC of transformed Ag. Likewise, for nano-ZnO, the form-specific approach reduced the mean RCR from 1.2 to 0.86. For nano-TiO2 , the form-specific assessment did not change the mean RCR of 0.026. This analysis shows that a form-specific approach can have an influence on the assessment of the environmental risks of ENMs and that, given the availability of form-specific release models, an updated ERA for ENMs can be performed. Environ Toxicol Chem 2021;40:2629-2639. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Single and combined toxicity of amino-functionalized polystyrene nanoparticles with potassium dichromate and copper sulfate on brine shrimp Artemia franciscana larvae.

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.

Ecotoxicity of silica nanoparticles in aquatic organisms: An updated review.

This review aims to (i) provide a current overview of the main characteristics of SiNP (physical and chemical properties, applications, and emissions), (ii) evaluate the scientific production up to date concerning SiNP, with focus on their toxic effects, through a bibliometric analysis, (iii) describe the main toxic mechanisms of SiNP, (iv) assess the current knowledge about ecotoxicity of SiNP on aquatic organisms (marine and freshwater), and (v) identify the main gaps in the knowledge of SiNP toxicity from an environmentally point of view. The scientific production of SiNP concerning their chemical and physical characteristics has increased exponentially. However, little information is available regarding their ecotoxicity. Particle functionalization is a key factor that reduces SiNP toxicity. Most of the studies employed standard species as test organisms, being the local/native ones poorly represented. Further studies employing long-term exposures and environmentally relevant concentrations are needed to deepen the knowledge about this emergent pollutant.

Comparative toxicity of ionic and nanoparticulate zinc in the species Cymodoce truncata, Gammarus aequicauda and Paracentrotus lividus.

Due to the continuous development, production and consumption of nanoparticles (NPs), their release, fate and effects in marine coastal environment can represent a major concern. The aim of this study was to evaluate the toxicity of ZnO nanoparticles (ZnO NPs) and compare it to bulk ZnSO4 on three macroinvertebrates: the isopod Cymodoce truncata (i.e. used for the first time in ecotoxicology), the amphipod Gammarus aequicauda and the sea urchin Paracentrotus lividus. This study showed concentration- and time-dependent relationships for all biological models for both ZnO NPs and ZnSO4. Both Zn forms elicited high toxicity to G. aequicauda and C. truncata juveniles, but ZnO NPs induced comparable responses to both species (96h-LC50 = 0.30 and 0.37 mg/L for G. aequicauda and C. truncata, respectively; p > 0.05), while differences were found after ZnSO4 exposure (96h-LC50 = 0.28 and 0.63 mg/L, respectively; p < 0.05). ZnO NPs generated sub-lethal effects on P. lividus embryos (72h-EC50 = 0.04 (0.03, 0.05) mg/L), not significantly different from ZnSO4 ones (72h-EC50 = 0.06 (0.05, 0.07) mg/L). Effects of ZnO NPs were similar to existing literature data for other testing species. C. truncata can be considered as a promising new biological model in (nano)ecotoxicology.

Confirmatory assays for transient changes of omics in soil invertebrates - Copper materials in a multigenerational exposure.

Environmental risk assessment (ERA) based on effects caused by chronic and longer term exposure is highly relevant. Further, if mechanistic based approaches (e.g. omics) can be included, beyond apical endpoints (e.g. reproduction), the prediction of effects increases. For Cu NMs (and CuCl2) this has been studied in detail, covering multi-omics and apical effects using the soil standard species Enchytraeus crypticus. The intermediate level effects like cell/tissue and organ alterations represent a missing link. In the present study we aimed to: 1) perform long term exposure to Cu materials (full life cycle and multigeneration, 46 and 224 days) to collect samples; 2) perform histology and immunohistochemistry on collected samples at 12 time points and 17 treatments; 3) integrate all levels of biological organization onto an adverse outcome pathway (AOP) framework. CuO NMs and CuCl2 caused both similar and different stress response, either at molecular initiating events (MIE) or key events (KEs) of higher level of biological organization. Cell/Tissue and organ level, post-transcriptional and transcriptional mechanisms, through histone modifications and microRNA related protein, were similarly affected. While both Cu forms affected the Notch signalling pathway, CuCl2 also caused oxidative stress. Different mechanisms of DNA methylation (epigenetics) were activated by CuO NMs and CuCl2 at the MIE.