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A suspension of copper oxide nanoparticles (CuO NPs) is a mixture of dissolved and particulate Cu, the relative proportions of which highly depend on the water chemistry. However, the relationship between different proportions of particulate and dissolved Cu and the overall toxicity of CuO NPs is still unknown. This study investigated the response of Chlorella vulgaris to CuO NPs at varying solution pH and at different tannic acid (TA) additions, with a focus on exploring whether and how dissolved and particulate Cu contribute to the overall toxicity of CuO NPs. The results of the exposure experiments demonstrated the involvement of both dissolved and particulate Cu in inducing toxicity of CuO NPs, and the inhibition of CuO NPs on cell density of Chlorella vulgaris was found to be significantly (p < 0.05) alleviated with increased levels of TA and pH (< 8). Using the independent action model, the contribution to toxicity of particulate Cu was found to be enhanced with increasing pH values and TA concentrations. The toxic unit indicator better (R 2 = 0.86, p < 0.001) explained impacts of CuO NPs on micro-algae cells than commonly used mass concentrations (R 2 = 0.27-0.77, p < 0.05) across different levels of pH and TA. Overall, our study provides an additivity-based method to improve the accuracy of toxicity prediction through including contributions to toxicity of both dissolved and particulate Cu and through eliminating the uneven distribution of data due to large variations in total Cu, particulate Cu, dissolved Cu, Cu 2+ activities, Cu-TA complexes and other Cu-complexes concentrations with varying water chemistry conditions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Research on theoretical prediction methods for the mixture toxicity of engineered nanoparticles (ENPs) faces significant challenges. The application of in silico methods based on machine learning is emerging as an effective strategy to address the toxicity prediction of chemical mixtures. Herein, we combined toxicity data generated in our lab with experimental data reported in the literature to predict the combined toxicity of seven metallic ENPs for Escherichia coli at different mixing ratios (22 binary combinations). We thereafter applied two machine learning (ML) techniques, support vector machine (SVM) and neural network (NN), and compared the differences in the ability to predict the combined toxicity by means of the ML-based methods and two component-based mixture models: independent action and concentration addition. Among 72 developed quantitative structure-activity relationship (QSAR) models by the ML methods, two SVM-QSAR models and two NN-QSAR models showed good performance. Moreover, an NN-based QSAR model combined with two molecular descriptors, namely enthalpy of formation of a gaseous cation and metal oxide standard molar enthalpy of formation, showed the best predictive power for the internal dataset (R 2 test  = 0.911, adjusted R 2 test  = 0.733, RMSE test  = 0.091, and MAE test  = 0.067) and for the combination of internal and external datasets (R 2 test  = 0.908, adjusted R 2 test  = 0.871, RMSE test  = 0.255, and MAE test  = 0.181). In addition, the developed QSAR models performed better than the component-based models. The estimation of the applicability domain of the selected QSAR models showed that all the binary mixtures in training and test sets were in the applicability domain. This study approach could provide a methodological and theoretical basis for the ecological risk assessment of mixtures of ENPs., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Herein, we investigated to which extent metallic nanoparticles (MNPs) affect the trophic transfer of other coexisting MNPs from lettuce to terrestrial snails and the associated tissue-specific distribution using toxicokinetic (TK) modeling and single-particle inductively coupled plasma mass spectrometry. During a period of 22 days, snails were fed with lettuce leaves that were root exposed to AgNO 3 (0.05 mg/L), AgNPs (0.75 mg/L), TiO 2 NPs (200 mg/L), and a mixture of AgNPs and TiO 2 NPs (equivalent doses as for single NPs). The uptake rate constants ( k u ) were 0.08 and 0.11 kg leaves/kg snail/d for Ag and 1.63 and 1.79 kg leaves/kg snail/d for Ti in snails fed with NPs single- and mixture-exposed lettuce, respectively. The elimination rate constants ( k e ) of Ag in snails exposed to single AgNPs and mixed AgNPs were comparable to the corresponding k u , while the k e for Ti were lower than the corresponding k u . As a result, single TiO 2 NP treatments as well as exposure to mixtures containing TiO 2 NPs induced significant biomagnification from lettuce to snails with kinetic trophic transfer factors (TTF k ) of 7.99 and 6.46. The TTF k of Ag in the single AgNPs treatment (1.15 kg leaves/kg snail) was significantly greater than the TTF k in the mixture treatment (0.85 kg leaves/kg snail), while the fraction of Ag remaining in the body of snails after AgNPs exposure (36%) was lower than the Ag fraction remaining after mixture exposure (50%). These results indicated that the presence of TiO 2 NPs inhibited the trophic transfer of AgNPs from lettuce to snails but enhanced the retention of AgNPs in snails. Biomagnification of AgNPs from lettuce to snails was observed in an AgNPs single treatment using AgNPs number as the dose metric, which was reflected by the particle number-based TTFs of AgNPs in snails (1.67, i.e., higher than 1). The size distribution of AgNPs was shifted across the lettuce-snail food chain. By making use of particle-specific measurements and fitting TK processes, this research provides important implications for potential risks associated with the trophic transfer of MNP mixtures.

The goal of the current study was to quantify the trophic transfer of copper nanoparticles (CuNPs) in a food chain consisting of the microalga Pseudokirchneriella subcapitata as the representative of primary producer, the grazer Daphnia magna, and the omnivorous mysid Limnomysis benedeni. To quantify the size and number concentration of CuNPs in the biota, tissue extraction with tetramethylammonium hydroxide (TMAH) was performed and quantification was done by single particle inductively coupled plasma mass spectrometry (sp-ICP-MS). The bioconcentration factor (BCF) of the test species for CuNPs varied between 10 2 - 10 3 L/kg dry weight when expressing the internal concentration on a mass basis, which was lower than BCF values reported for Cu 2+ (10 3 - 10 4 L/kg dry weight). The particle size of CuNPs determined by sp-ICP-MS ranged from 22 to 40 nm in the species. No significant changes in the particle size were measured throughout the food chain. Moreover, the measured number of CuNPs in each trophic level was in the order of 10 13 particles/kg wet weight. The calculated trophic transfer factor (mass concentration basis) was > 1. This indicates biomagnification of particulate Cu from P. subcapitata to L. benedeni. It was also found that the uptake of particulate Cu (based on the particle number concentration) was mainly from the dietary route rather than from direct aqueous exposure. Furthermore, dietary exposure to CuNPs had a significant effect on the feeding rate of mysid during their transfer from daphnia to mysid and from alga through daphnia to mysid. This work emphasizes the importance of tracing the particulate fraction of metal-based engineered nanoparticles when studying their uptake and trophic transfer., (Copyright © 2022. Published by Elsevier Inc.)

Many host-microbiota interactions depend on the recognition of microbial constituents by toll-like receptors of the host. The impacts of these interactions on host health can shape the hosts response to environmental pollutants such as nanomaterials. Here, we assess the role of toll-like receptor 2 (TLR2) signaling in the protective effects of colonizing microbiota against silver nanoparticle (nAg) toxicity to zebrafish larvae. Zebrafish larvae were exposed to nAg for two days, from 3 to 5 days post-fertilization. Using an il1ß-reporter line, we first characterized the accumulation and particle-specific inflammatory effects of nAg in the total body and intestinal tissues of the larvae. This showed that silver gradually accumulated in both the total body and intestinal tissues, yet specifically caused particle-specific inflammation on the skin of larvae. Subsequently, we assessed the effects of microbiota-dependent TLR2 signaling on nAg toxicity. This was done by comparing the sensitivity of loss-of-function zebrafish mutants for TLR2, and each of the TLR2-adaptor proteins MyD88 and TIRAP (Mal), under germ-free and microbially-colonized conditions. Irrespective of their genotype, microbially-colonized larvae were less sensitive to nAg than their germ-free siblings, supporting the previously identified protective effect of microbiota against nAg toxicity. Under germ-free conditions, tlr2, myd88 and tirap mutants were equally sensitive to nAg as their wildtype siblings. However, when colonized by microbiota, tlr2 and tirap mutants were more sensitive to nAg than their wildtype siblings. The sensitivity of microbially-colonized myd88 mutants did not differ significantly from that of wildtype siblings. These results indicate that the protective effect of colonizing microbiota against nAg-toxicity to zebrafish larvae involves TIRAP-dependent TLR2 signaling. Overall, this supports the conclusion that host-microbiota interactions affect nanomaterial toxicity to zebrafish larvae., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Similarity assessment is one of the means of optimally using scarcely available experimental data on the fate and hazards of nanoforms (NFs) for regulatory purposes. For a set of NFs that are shown to be similar it is allowed in a regulatory context to apply the information available on any of the NFs within the group to the whole set of NFs. Obviously, a proper justification for such a similarity assessment is to be provided. Within the context of exemplifying such a justification, a case study was performed aimed at assessing the similarity of a set of spherical metallic NFs that different with regard to chemical composition (three metals) and particle size (three different sizes). The endpoints of assessment were root elongation and biomass increase of lettuce (Lactuca sativa L.) seedlings and exposure assessment was performed in order to express the actual exposure concentration in terms of time-weighted average particle concentrations. The results of the study show that for the specific endpoints assessed, chemical composition is driving NF toxicity and this is mostly due to impacts on the fate of the NFs. On the other hand, particle size of Cu NFs had a negligible impact on the dose-response relationships for the specific endpoints assessed. It is thus concluded that hazard data available on spherical Cu NF tested in our case can be used to inform on the hazards of any spherical Cu NF within the size range of 25-100 nm, but only applies for the certain endpoints. Also, toxicity data for the Cu 2+ -ion are suited for such a similarity assessment., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Various modern products have metallic nanoparticles (MNPs) embedded to enhance products performance. Technological advances enable nowadays even multiple hybrid nanoparticles. Consequently, the future co-release of multiple MNPs will inevitably result in the presence of MNP mixtures in the environment. An important question is if the responses of mixtures of MNPs can be dealt with in a similar way as with the responses of biota to mixtures of metal salts. Moreover, natural organic matter (NOM) is an important parameter affecting the behavior and effect of MNPs. Herein, we determined the joint toxicity and accumulation of copper nanoparticles (CuNPs) and zinc oxide nanoparticles (ZnONPs) in Daphnia magna in the absence and presence of Suwannee River natural organic matter (SR-NOM), compared to the joint toxicity and accumulation of corresponding metal salts. The results of toxicity testing showed that the joint toxicity of CuNPs + ZnONPs was greater than the single toxicity of CuNPs or ZnONPs. The joint toxic action of CuNPs + ZnONPs was additive or more-than-additive for D. magna. A similar pattern was found in the toxicity of the mixtures of Cu- and Zn-salts from the literature data. The presence of SR-NOM had no significant impact on the joint toxicity of CuNPs + ZnONPs. The calculated component-specific contribution to overall toxicity indicated that SR-NOM increased the relative contribution of dissolved ions released from the MNPs to the toxicity of the binary mixtures at high-effect concentrations of individual MNPs. Moreover, dissolved Zn-ions released from the ZnONPs were found to dominate the joint toxicity of CuNPs + ZnONPs in the presence of SR-NOM. Furthermore, the results of the accumulation experiment displayed that the presence of SR-NOM significantly enhanced the accumulation of either CuNPs or ZnONPs in D. magna exposed to the MNP mixtures., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The increasing application of biosolids and agrochemicals containing silver nanoparticles (AgNPs) and titanium dioxide nanoparticles (TiO 2 NPs) results in their inevitable accumulation in soil, with unknown implications along terrestrial food chains. Here, the trophic transfer of single NPs and a mixture of AgNPs and TiO 2 NPs from lettuce to snails and their associated impacts on snails were investigated. Both AgNPs and TiO 2 NPs were transferred from lettuce to snails with trophic transfer factors (defined as the ratio of the Ag/Ti concentration in snail tissues to the Ag/Ti concentration in lettuce leaves) of 0.2-1.1 for Ag and 3.8-47 for Ti. Moreover, the majority of Ag captured by snails in the AgNP-containing treatments was excreted via feces, whereas more than 70% of Ti was distributed in the digestive gland of snails in the TiO 2 NP-containing treatments. Additionally, AgNP-containing treatments significantly inhibited the activity of snails, while TiO 2 NP-containing treatments significantly reduced feces excretion of snails. Furthermore, the concurrent application of AgNPs and TiO 2 NPs did not affect the biomagnification and distribution patterns of Ag and Ti in snails, whereas their co-existence exhibited more severe inhibition of the growth and activity of snails than in the case of applying AgNPs or TiO 2 NPs alone. This highlights the possibility of nanoparticle transfer to organisms of higher trophic levels via food chains and the associated risks to ecosystem health.

With the expanding nanotechnology, nanoparticles (NPs) embedded products are used in the agricultural sector to improve soil fertility. Thus, two typical metal oxides NPs and their mixtures were applied in different doses to evaluate the impacts on soil microbes. CuO and ZnO NPs boosted soil microbial communities as reflected by the increased number of extractable bacterial or fungal groups and the enlarged values of Chao 1, ACE, and Shannon indices. Relative abundance of some susceptible taxa such as Sphingomonadales increased with increasing concentrations of ZnO NPs, while IMCC26256 decreased with increasing concentrations of CuO NPs. The mixture of CuO and ZnO NPs did not show more promotional effects on the soil bacterial community than the sum of individual effects. Increased soil organic carbon mitigated the impacts on soil bacteria for CuO NPs, but not for ZnO NPs. As micro-nutrients, the ions released from CuO and ZnO NPs had the potential to promote soil microbial community richness and diversity. However, the positive impacts of MNPs were impaired at dosage higher than 250 mg kg -1 soil (213.08 mg kg -1 soil of Cu, 162.73 mg kg -1 soil of Zn). Thus, the application dose and soil type other than the coexistence of MNPs should be considered before the wide use in increasing agricultural productivity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Metal-based nanoparticles exhibiting antimicrobial activity are of emerging concern to human and environmental health. In addition to their direct adverse effects to plants and animals, indirect effects resulting from disruption of beneficial host-microbiota interactions may contribute to the toxicity of these particles. To explore this hypothesis, we compared the acute toxicity of silver and zinc oxide nanoparticles (nAg and nZnO) to zebrafish larvae that were either germ-free or colonized by microbiota. Over two days of exposure, germ-free zebrafish larvae were more sensitive to nAg than microbially colonized larvae, whereas silver ion toxicity did not differ between germ-free and colonized larvae. Using response addition modeling, we confirmed that the protective effect of colonizing microbiota against nAg toxicity was particle-specific. Nearly all mortality among germ-free larvae occurred within the first day of exposure. In contrast, mortality among colonized larvae increased gradually over both exposure days. Concurrent with this gradual increase in mortality was a marked reduction in the numbers of live host-associated microbes, suggesting that bactericidal effects of nAg on protective microbes resulted in increased mortality among colonized larvae over time. No difference in sensitivity between germ-free and colonized larvae was observed for nZnO, which dissolved rapidly in the exposure medium. At sublethal concentrations, these particles moreover did not exert detectable bactericidal effects on larvae-associated microbes. Altogether, our study shows the importance of taking host-microbe interactions into account in assessing toxic effects of nanoparticles to microbially colonized hosts, and provides a method to screen for microbiota interference with nanomaterial toxicity.

Whether toxicity of silver nanoparticles (AgNPs) to organisms originates from the nanoparticles themselves or from the dissolved Ag-ions is still debated, with the majority of studies claiming that extracellular release of Ag-ions is the main cause of toxicity. The objective of this study was to determine the contributions of both particles and dissolved ions to toxic responses, and to better understand the underlying mechanisms of toxicity. In addition, the pathways of AgNPs exposure to plants might play an important role and therefore are explicitly studied as well. We systematically assessed the phytotoxicity, internalization, biodistribution, and antioxidant responses in lettuce (Lactuca sativa) following root or foliar exposure to AgNPs and ionic Ag at various concentrations. For each endpoint the relative contribution of the particle-specific versus the ionic form was quantified. The results reveal particle-specific toxicity and uptake of AgNPs in lettuce as the relative contribution of particulate Ag accounted for more than 65% to the overall toxicity and the Ag accumulation in whole plant tissues. In addition, particle toxicity is shown to originate from the accumulation of Ag in plants by blocking nutrient transport, while ion toxicity is likely due to the induction of excess ROS production. Root exposure induced higher toxicity than foliar exposure at comparable exposure levels. Ag was found to be taken up and subsequently translocated from the exposed parts of plants to other portions regardless of the exposure pathway. These findings suggest particle related toxicity, and demonstrate that the accumulation and translocation of silver nanoparticles need to be considered in assessment of environmental risks and of food safety following consumption of plants exposed to AgNPs by humans., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Humic substances are the dominant dissolved organic matter fraction in the aqueous phase of environmental media. They would inevitably react with chemicals released into the environment. The influence of dissolved humic acid (DHA) on the dissolution and dispersion of copper oxide nanoparticles (CuO NPs, 50 nm, 49.57 mg L -1 ) was therefore investigated in the present study. In addition to dispersing CuO NPs and reducing the size of the aggregates, the amount of released Cu from CuO NPs was found to increase over time with increasing concentrations of DHA, 96% of which was present as organic complexes after 72 h. At DHA concentrations exceeding 16.09 mg C L -1 , the complexation coefficients of DHA with Cu and the adsorptivity of CuO NPs to DHA were both reduced due to increased homo-conjugation of DHA as promoted by negative charge-assisted H-bond. Although the adsorption capacity of DHA kept increasing up to 57.07 mg C L -1 , the hydrodynamic diameter and ζ-potential were similar and the percentages of total released Cu continued to increase linearly to 4.92% at higher levels of DHA (30.13-57.07 mg C L -1 ). Thereupon, DHA promoted the dissolution of CuO NPs in a concentration-dependent fashion. The driving force was complexation of Cu by DHA, rather than the balancing between the exposed and the covered surface area of the CuO NPs due to DHA adsorption. Our findings facilitate understanding the underlying mechanisms on how DHA impacts the CuO NPs environmental behavior (or fate) as well as on their kinetics., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

We systematically investigated how the combinations of size, shape and the natural organic matter (NOM)-ecocorona of gold (Au) engineered nanoparticles (ENPs) influence the attachment of the particles to algae and physical toxicity to the cells. Spherical (10, 60 and 100 nm), urchin-shaped (60 nm), rod-shaped (10 × 45, 40 × 60 and 50 × 100 nm), and wire-shaped (75 × 500, 75 × 3000 and 75 × 6000 nm) citrate-coated and NOM-coated Au-ENPs were used. Among the spherical particles only the spherical 10 nm Au-ENPs caused membrane damage to algae. Only the rod-shaped 10 × 45 nm induced membrane damage among the rod-shaped Au-ENPs. Wire-shaped Au-ENPs caused no membrane damage to the algae. NOM ecocorona decreased the membrane damage effects of spherical 10 nm and rod-shaped 10 × 45 nm ENPs. The spherical Au-ENPs were mostly loosely attached to the cells compared to other shapes, whereas the wire-shaped Au-ENPs were mostly strongly attached compared to particles with other shapes. NOM ecocorona determined the strength of Au-ENPs attachment to the cell wall, leading to the formation of loose rather than strong attachment of Au-ENPs to the cells. After removal of the loosely and strongly attached Au-ENPs, some particles remained anchored to the surface of the algae. The highest concentration was detected for spherical 10 nm Au-ENPs followed by rod-shaped 10 × 45 nm Au-ENPs, while the lowest concentration was observed for the wire-shaped Au-ENPs. The combined effect of shape, size, and ecocorona controls the Au-ENPs attachment and physical toxicity to cells.

Some metal-based engineered nanoparticles (ENPs) undergo fast dissolution and/or aggregation when they are released in the environment. The underlying processes are controlled by psychochemical/biological parameters of the environment and the properties of the particles. In this study, we investigated the interaction between algal cells and zero valent copper nanoparticles (Cu 0 -ENPs) to elucidate how the cells influence the dissolution and aggregation kinetics of the particles and how these kinetics influence the cellular uptake of Cu. Our finding showed that the concentration of dissolved Cu ([Cu] dissolved ) in the supernatant of the culture media without algal cells was higher than the [Cu] dissolved in the media with algal cells. In the absence of the cells, dissolved organic matter (DOC) increased the dissolution of the particle due to increasing the stability of the particles against aggregation, thus increasing the available surface area. In the presence of algae, Cu 0 -ENPs heteroaggregated with the cells. Thus, the available surface area decreased over time and this resulted in a low dissolution rate of the particles. The DOC corona on the surface of the particles increased the heteroaggregation of the particles with the cells and decreases the uptake of the particles. Our findings showed that microorganisms influence the fate of ENPs in the environment, and they do so by modifying the dissolution and aggregation kinetics of the Cu 0 -ENPs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The combined effects of pH, dissolved organic carbon (DOC) and Ca 2+ /Mg 2+ on the dissolution and aggregation kinetics of zero valent copper engineered nanoparticles (Cu 0 ENPs) were investigated. The dissolution and aggregation of the particles were studied in (a) synthetic aqueous media, similar in chemistry to natural surface waters, and (b) natural surface waters samples, for up to 32 or 24 h. The DOC stabilized the particles and prevented aggregation, and thus increased the available surface area. The higher available surface area in turn accelerated the dissolution of the particles. The presence of Ca 2+ /Mg 2+ , however, changed the aggregation and the dissolution of the DOC-stabilized particles. The influence of Ca 2+ /Mg 2+ on DOC-stabilized particles was different at different pH's. In the absence of DOC, 10 mM of Ca 2+ /Mg 2+ induced charge reversal on the particles and caused particle stability against aggregation. This subsequently increased particles dissolution. The results obtained with regard to dissolution and aggregation of the particles in natural surface waters were compared with those determined for the synthetic waters. This comparison showed that the behavior of the particles in the natural surface waters was mostly similar to the behavior determined for media at pH 9. Overall, the current study provides some novel insights into the simultaneous effects of physicochemical parameters of water on particle stability against aggregation and dissolution, and provides data about how the processes of aggregation and dissolution of Cu 0 ENPs interact and jointly determine the overall particle fate., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Important questions raised in (nano)ecotoxicology are whether biodistribution of nanoparticles (NPs) is affected by particle shape and to what extent local adverse responses are subsequently initiated. For nanomedicine, these same questions become important when the labeled NPs lose the labeling. In this study, we investigated the biodistribution patterns of gold nanoparticles (AuNPs) as well as immune-related local and systemic sublethal markers of exposure and behavioral assessment. Hatched zebrafish embryos were exposed to four differently shaped non-coated AuNPs with comparable sizes: nanospheres, nanorods, nano-urchins, and nano-bipyramids. Shape-dependent trafficking of the particles resulted in a different distribution of the particles over the target organs. The differences across the distribution patterns indicate that the particles behave slightly different, although they eventually reach the same target organs - yet in different ratios. Mainly local induction of the immune system was observed, whereas systemic immune responses were not clearly visible. Macrophages were found to take AuNPs from the body fluid, be transferred into the veins and transported to digestive organs for clearance. No significant behavioral toxicological responses in zebrafish embryos were observed after exposure. The trafficking of the particles in the macrophages indicates that the particles are removed via the mononuclear phagocytic system. The different ratios in which the particles are distributed over the target organs indicate that the shape influences their behavior and eventually possibly the toxicity of the particles. The observed shape-dependent biodistribution patterns might be beneficial for shape-specific targeting in nanomedicine and stress the importance of incorporating shape-features in nanosafety assessment.

A multistep sample preparation method was developed to separate metal-based engineered nanoparticles (ENPs) from biological samples. The method was developed using spiked zebrafish tissues and standard titanium dioxide (TiO 2 ) and cerium dioxide (CeO 2 ) ENPs. Single-particle inductively coupled plasma mass spectrometry was used to quantify the separated particles in terms of number concentration. This method demonstrated mass recoveries of more than 90% and did not strikingly alter the median particles size. High number recoveries were calculated for CeO 2 ENPs (>84%). Particle number recoveries were poor for TiO 2 ENPs (<25%), which could be due to the interference of 48 Ca with the measured isotope 48 Ti. The method was verified using zebrafish exposed to CeO 2 ENPs to test its applicability for nanotoxicokinetic investigations. Total mass of Ce and particle number concentration of CeO 2 ENPs were measured in different tissues. Notably, the mass-based biodistribution of Ce in the tissues did not follow the number-based biodistribution of CeO 2 . Moreover, the calculated mass-based bioconcentration factors showed a different pattern in comparison to the number-based bioconcentration factors. Our findings suggest that considering mass as the sole dose-metric may not provide sufficient information to investigate toxicity and toxicokinetics of ENPs.

A full-factorial test design was applied to systematically investigate the contribution and significance of water chemistry parameters (pH, divalent cations and dissolved organic carbon (DOC) concentration) and their interactions on the behavior and fate of copper nanoparticles (CuNPs). The total amount of Cu remaining in the water column after 48 h of incubation was mostly influenced by divalent cation content, DOC concentration and the interaction of divalent cations and DOC. DOC concentration was the predominant factor influencing the dissolution of CuNPs, which was far more important than the effect of pH in the range from 6 to 9 on the dissolution of the CuNPs. The addition of DOC at concentrations ranging from 5 to 50 mg C/L resulted in a 3-5 fold reduction of dissolution of CuNPs after 48 h of incubation, as compared to the case without addition of DOC. Divalent cation content was found to be the most influential factor regarding aggregation behavior of the particles, followed by DOC concentration and the interaction of divalent cations and DOC. In addition, the aggregation behavior of CuNPs rather than particulate dissolution explained most of the variance in the sedimentation profiles of CuNPs. These results are meaningful for improved understanding and prediction of the behavior and fate of metallic NPs in aqueous environments., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

A species sensitivity distribution (SSD) for engineered nanomaterials (ENMs) ranks the tested species according to their sensitivity to a certain ENM. An SSD may be used to estimate the maximum acceptable concentrations of ENMs for the purpose of environmental risk assessment. To construct SSDs for metal-based ENMs, more than 1800 laboratory derived toxicity records of metallic ENMs from >300 publications or open access scientific reports were retrieved. SSDs were developed for the metallic ENMs grouped by surface coating, size, shape, exposure duration, light exposure, and different toxicity endpoints. It was found that PVP- and sodium citrate- coatings enhance the toxicity of Ag ENMs as concluded from the relevant SSDs. For the Ag ENMs with different size ranges, differences in behavior and/or effect were only observed at high exposure concentrations. The SSDs of Ag ENMs separated by both shape and exposure duration were all nearly identical. Crustaceans were found to be the most vulnerable group to metallic ENMs. In spite of the uncertainties of the results caused by limited data quality and availability, the present study provided novel information about building SSDs for distinguished ENMs and contributes to the further development of SSDs for metal-based ENMs., (Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Toxicity of metallic nanoparticle suspensions (NP (total) ) is generally assumed to result from the combined effect of the particles present in suspensions (NP (particle) ) and their released ions (NP (ion) ). Evaluation and consideration of how water chemistry affects the particle-specific toxicity of NP (total) are critical for environmental risk assessment of nanoparticles. In this study, it was found that the toxicity of Cu NP (particle) to Daphnia magna, in line with the trends in toxicity for Cu NP (ion) , decreased with increasing pH and with increasing concentrations of divalent cations and dissolved organic carbon (DOC). Without the addition of DOC, the toxicity of Cu NP (total) to D. magna at the LC50 was driven mainly by Cu NP (ion) (accounting for ≥53% of the observed toxicity). However, toxicity of Cu NP (total) in the presence of DOC at a concentration ranging from 5 to 50mg C/L largely resulted from the NP (particle) (57%-85%), which could be attributable to the large reduction of the concentration of Cu NP (ion) and the enhancement of the stability of Cu NP (particle) when DOC was added. Our results indicate that water chemistry needs to be explicitly taken into consideration when evaluating the role of NP (particle) and NP (ion) in the observed toxicity of NP (total) ., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The increasing number of applications in which copper oxide nanoparticles (CuO NPs) are used, may lead to potential release of CuO NPs into the environment. However, the impact of natural organic matters on the behavior and fate of CuO NPs in aquatic media is still largely unknown. In this study, the dissolution and aggregation of CuO NPs under the exposure of tannic acid (TA) were monitored over a period of 72 h, with a focus on assessing the contributions of solution pH changes and complexation reactions. Results showed that the total amount of Cu 2+ released from CuO NPs increased in the presence of TA especially at the highest TA concentration of 73.5 μmol/L. Although TA was observed to wrap around the CuO NPs, the aggregation of CuO NPs was not strongly influenced by TA and by the solution pH as investigated in this study. The kinetics of Cu 2+ release were fitted using the modified pseudo second-order model and the rate of dissolution was assessed to be highest at TA = 14.7 μmol/L. At pH = 4, the increased H + concentration was responsible for increased Cu 2+ release, whereas the complexation reaction between Cu 2+ and TA dominated at pH = 7. These findings suggested that the effects of TA on the dissolution of CuO NPs were a combination of solution pH change and complexation reaction, the relative fractions of which also depended on the solution pH. Additionally, the percentage of Cu 2+ released from the CuO NPs was found to increase upon decreasing concentrations of CuO NPs. Our work helps to further understand how and to which extent natural organic matters affect the behavior and fate of CuO NPs., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Metallic nanoparticles (NPs) differ from other metal forms with respect to their large surface to volume ratio and subsequent inherent reactivity. Each new modification to a nanoparticle alters the surface to volume ratio, fate and subsequently the toxicity of the particle. Newly-engineered NPs are commonly available only in low quantities whereas, in general, rather large amounts are needed for fate characterizations and effect studies. This challenge is especially relevant for those NPs that have low inherent toxicity combined with low bioavailability. Therefore, within our study, we developed new testing strategies that enable working with low quantities of NPs. The experimental testing method was tailor-made for NPs, whereas we also developed translational models based on different dose-metrics allowing to determine dose-response predictions for NPs. Both the experimental method and the predictive models were verified on the basis of experimental effect data collected using zebrafish embryos exposed to metallic NPs in a range of different chemical compositions and shapes. It was found that the variance in the effect data in the dose-response predictions was best explained by the minimal diameter of the NPs, whereas the data confirmed that the predictive model is widely applicable to soluble metallic NPs. The experimental and model approach developed in our study support the development of (eco)toxicity assays tailored to nano-specific features., Competing Interests: The authors declare no conflict of interest. The founding sponsors had no role in the design of the study; neither in the collection, analysis or interpretation of data, nor in the writing of the manuscript or the decision to publish the results.

The mean primary diameter (d) of nanoparticles (NPs) is commonly used as the best basis to assess the dose metric for expressing the toxicity of spherical NPs. However, d bears no relevance for nonspherical NPs. In the present study, the mean surface area to volume ratio, applicable to both spherical and nonspherical NPs, was used to replace d given the aim of obtaining the best dose metric (volume, surface area, or number) of differently shaped metallic NPs in vivo (9 organisms) and in vitro (6 mammalian cell lines). The slope of the curves obtained by relating the total particle number of NPs at various effect concentrations to the mean surface area to volume ratio was subsequently used to deduce the best dose metric. For the majority of the organisms studied, it was found that NP volume is the most appropriate dose metric, independent of the composition of the NPs tested. For 3 organisms exposed to Ag NPs, however, surface area was found to be the best dose metric. It is therefore concluded that the optimum dose metric depends on organism and NP properties. Environ Toxicol Chem 2016;35:2466-2473. © 2016 SETAC., (© 2016 SETAC.)

Although the risks of metallic nanoparticles (NPs) to aquatic organisms have already been studied for >10years, our understanding of the link between the fate of particles in exposure medium and their toxicity is still in its infancy. Moreover, most of the earlier studies did not distinguish the contribution of particles and soluble ions to the toxic effects caused by suspensions of metallic NPs. In this study, the toxicity of CuNPs to Daphnia magna upon modification of the exposure conditions, achieved by aging the suspensions of CuNPs and by altering water chemistry parameters like the pH and levels of dissolved organic carbon (DOC), was investigated. The LC50 values for CuNPs exposure decreased by about 30% after 7days of aging. The LC50 values increased >12-fold upon addition of DOC at concentrations ranging from 0 to 10mg/L to the exposure medium. Changing the pH from 6.5 to 8.5 resulted in a 3-fold higher LC50 value. Furthermore, it was found that during 7days of aging of the exposure medium (without addition of DOC and at pH7.8), the toxicity could be mostly ascribed to the particles present in the suspension (around 70%). However, adding DOC or decreasing the pH of the exposure medium reduced the contribution of the particles to the observed toxicity. We thus found that the effective concentration regarding the toxicity was mainly driven by the contribution of the soluble ions in the presence of DOC or at pH6.5. Our results suggest that the toxicity results of CuNPs obtained from laboratory tests may overestimate the risk of the particles in polluted waters due to the common absence of DOC in laboratory test solutions. Moreover, the role of the ions shedding from CuNPs is very important in explaining the toxicity in natural waters., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Copper nanoparticles (CuNPs) are used extensively in a wide range of products and the potential for toxicological impacts in the aquatic environment is of high concern. In this study, the fate and the acute toxicity of spherical 50nm copper nanoparticles was assessed in juvenile rainbow trout (Oncorhynchus mykiss), fathead minnow (Pimephales promelas) and zebrafish (Danio rerio) for in vivo aqueous exposures following standardized OECD 203 guideline tests. The fate of the CuNPs in the aqueous media was temperature dependent. At the higher study temperature (26±1°C), there was both an enhanced particle aggregation and higher rate of dissolution compared with that at the lower study temperature (15±1°C). 96h LC50s of the CuNPs were 0.68±0.15, 0.28±0.04 and 0.22±0.08mg Cu/L for rainbow trout, fathead minnow and zebrafish, respectively. The 96h lowest-observed-effect concentration (LOEC) for the CuNPs were 0.17, 0.023 and <0.023mg/L for rainbow trout, fathead minnow, and zebrafish respectively, and are below the predicted environmental concentration of CuNPs for some aquatic environments suggesting a possible ecotoxicological risk to fish. Soluble copper was one of main drivers for the acute toxicity of the copper nanoparticles suspensions. Both CuNPs suspension and copper nitrate caused damage to gill filaments and gill pavement cells, with differences in sensitivity for these effects between the fish species studied. We show therefore common toxicological effects of CuNPs in different fish species but with differences in sensitivity with implications for hazard extrapolation between fish species., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This review establishes an inventory of existing toxicity data on nanoparticles (NPs) with the purpose of developing (Quantitative) Structure-Activity Relationships for NPs (nano-[Q]SARs), and also of maximising the use of scientific sources for NP risk assessment. From a data search carried out on 27 February 2014, a total of 910 publications were retrieved from the Web of Science™ Core Collection, and a database comprising 886 records of toxicity endpoints, based on these publications, was built. The test organisms mainly comprised bacteria, algae, yeast, protozoa, nematoda, crustacea and fish. The NPs consisted mostly of metals, metal oxides, nanocomposites and quantum dots. The data were analysed further, in order to: a) categorise each toxicity endpoint and the biological effects triggered by the NPs; b) survey the characterisation of the NPs used; and c) assess whether the data were suitable for nano-(Q)SAR development. Despite the efforts of numerous scientific programmes on nanomaterial safety and design, our study concluded that lack of data consistency prevents the use of experimental data in developing and validating nano-(Q)SARs. Finally, an outlook on the future of nano-(Q)SAR development is provided., (2015 FRAME.)

As a result of ever increasing applications, nanoparticles will eventually end up in the environment. However, currently no common principle has been established to help understand the toxicity of nanoparticles (NPs) across species. Therefore, it is difficult to estimate the potential risks of nanoparticles to untested species in the environment. The authors exposed 4 different sizes of copper nanoparticles (CuNPs) and 1 submicron-sized copper particle to 5 cladoceran species (Daphnia magna, Daphnia pulex, Daphnia galeata, Ceriodaphnia dubia, and Chydorus sphaericus) to investigate whether morphological attributes of species can help to assess the acute toxicity of CuNPs across species. The results showed that rod-shaped CuNPs caused much lower toxicity to all species than spherical CuNPs. Both the particles and ions contributed to the total toxicity of the CuNP suspensions. Moreover, the toxicity caused by particles in 5 different copper suspensions increases with decreasing body length, surface area, and body volume of neonates of 5 cladoceran species. Especially the correlations between body volume of the 5 cladoceran species tested and the corresponding toxicity caused by 5 different CuNPs were statistically significant, and in all cases radj (2) was higher than 0.51 (p < 0.001). The highest correlation was found between body volume and the toxicity of the 78-nm CuNPs (radj (2)  = 0.95, p < 0.001). To conclude, the correlations between attributes of cladoceran species and the toxicity of CuNPs reported in the present study evoke the possibility to assess and extrapolate the toxicity of nanoparticles across species with similar attributes., (© 2015 SETAC.)

Metallic nanoparticles can end up in aquatic ecosystems due to their widespread application. Even though the toxicological effects of metallic nanoparticles to a diversity of species have been reported extensively, the toxicological data achieved in different studies are not always comparable and little is known regarding the comparative toxicity of nanoparticles across species, as different test strategies and endpoints were applied. To attempt to fill this knowledge gap, Spirodela polyrhiza, Lemna minor and Wolffia arrhiza were exposed to 25 nm spherical copper nanoparticles to investigate the inhibiting effect of copper nanoparticle suspensions across species at three endpoints: total frond area, frond number and dry weight based relative growth rate. The total frond area based relative growth rate was found to be the most sensitive endpoint, with an EC50 of 1.15±0.09 mg/L for S. polyrhiza, 0.84±0.12 mg/L for L. minor and 0.64±0.05 mg/L for W. arrhiza. Both the particles and the copper ions contributed to the inhibiting effects of copper nanoparticle suspensions at all endpoints studied. Dose-response related inhibiting effects caused by the copper ions were found at all endpoints studied, whereas the particles only showed dose-response related inhibiting effects on the total frond area based relative growth rate. This suggests that different physiological processes are involved in case of exposure to particles and copper ions. W. arrhiza was found to be the most sensitive species tested and S. polyrhiza was the least sensitive species tested, when the inhibiting effect was evaluated based on the relative growth rate calculated from total frond area. These findings exemplify the importance of identifying the suitable endpoints of toxicity assessment and considering the intrinsic differences between species when evaluating the toxicological profile of metallic nanoparticles across species., (Copyright © 2015 Elsevier B.V. All rights reserved.)

There is increasing recognition that the wide use of nanoparticles, such as Cu (CuNPs) and ZnO nanoparticles (ZnONPs), may pose risks to the environment. Currently there is insufficient insight in the contribution of metal-based nanoparticles and their dissolved ions to the overall toxicity and accumulation. To fill in this gap, we combined the fate assessment of CuNPs and ZnONPs in aquatic test media with the assessment of toxicity and accumulation of ions and particles present in the suspensions. It was found that at the LC50 level of Daphnia magna exposed to the nanoparticle suspensions, the relative contributions of ions released from CuNPs and ZnONPs to toxicity were around 26% and 31%, respectively, indicating that particles rather than the dissolved ions were the major source of toxicity. It was additionally found that at the low exposure concentrations of CuNPs and ZnONPs (below 0.05 and 0.5 mg/L, respectively) the dissolved ions were predominantly accumulated, whereas at the high exposure concentrations (above 0.1 mg/L and 1 mg/L, respectively), particles rather than the released ions played a dominant role in the accumulation process. Our results thus suggest that consideration on the contribution of dissolved ions to nanoparticle toxicity needs to be interpreted with care.

A general approach is proposed that allows for quantifying the relative toxic contribution of ions released from metallic nanoparticles and of the particles themselves, as exemplified for the case of differently shaped zinc oxide (ZnO) nanoparticles (NPs) exposed to zebrafish embryos. First of all, the toxicity of suspensions of ZnO nanoparticles (NP(total))--nanospheres, nanosticks, cuboidal submicron particles (SMPs), and Zn(NO3)2--to the embryos was assessed. The observed toxicity of ZnO NP(total) is assumed to result from the combined effect of the particles present in the suspensions (NP(particle)) and of the dissolved Zn(2+) ions released from the particles (NP(ion)). Different addition models were used to explicitly account for the toxicity of NP(particle). The median lethal concentrations (LC50) of NP(particle) of nanospheres, nanosticks, and SMPs were found to range between 7.1 mg Zn/L and 11.9 mg Zn/L (i.e., to differ by a factor of 1.7). Behavioral performance showed no significant differences among all types of the NP(particle). The median effective concentrations (EC50) of the particles were found to range between 1.0 mg Zn/L and 2.2 mg Zn/L. At the LC50 of each particle suspension, the main contribution to lethality to zebrafish embryos was from the NP(particle) (52%-72%). For hatching inhibition, the NP(particle) was responsible for 38% to 83% of the adverse effects observed. The ZnO nanosticks were more toxic than any of the other NPs with regard to the endpoints mortality and hatching inhibition. The main contribution to toxicity to zebrafish embryos was from the NP(particle) at the LC50 and EC50 of each particle suspension., (© 2014 SETAC.)

Three sizes of copper nanoparticles (Cu NPs; 25 nm, 50 nm, and 100 nm), 1 submicron-sized particle, and Cu(NO3 )2 were added to the culture buffer of zebrafish embryos from 24 h postfertilization to 120 h postfertilization. In suspensions of Cu NPs and the Cu submicron-sized particle, the main contribution to the toxicity to zebrafish embryos was from the particle form of Cu particles (Cu NPparticle , >71%) rather than from dissolved Cu from the Cu particles (Cu NPion ). All particles tested as well as copper nitrate inhibited hatching, altered behavioral responses, and increased the incidence of malformations. Different kinds of abnormalities were observed in the morphology and behavior of the zebrafish embryos, depending on the particle size of the Cu suspensions tested. The median lethal concentrations of Cu NPparticle (25 nm, 50 nm, and 100 nm), the submicron-sized particle, and copper nitrate were 0.58 mg/L, 1.65 mg/L, 1.90 mg/L, 0.35 mg/L, and 0.70 mg/L, respectively. Submicron-sized particles and copper nitrate were more toxic than Cu NPs, and smaller Cu NPs were more toxic than larger Cu NPs. Dissolution of Cu NPs and the subsequent ion toxicity was not the primary mechanism of Cu NP toxicity in zebrafish embryos., (© 2014 SETAC.)

The four copper nanoparticles (CuNPs) with the size of 25, 50, 78 and 100 nm and one type of micron-sized particles (MPs) (~500 nm) were exposed to two mammalian (H4IIE and HepG2) and two piscine (PLHC-1 and RTH-149) cell lines to test the species-specific toxicities of CuNPs. The results showed that the morphologies, ion release and size of the particles all played an important role when investigating the toxicity. Furthermore, the authors found that the particle forms of CuNPs in suspensions highly contribute to the toxicity in all exposed cell lines whereas copper ions (Cu(2+)) only caused significant responses in mammalian cell lines, indicating the species-specific toxicity of CuNPs. This study revealed that the morphologies, ion release rate of NPs as well as the species-specific vulnerabilities of cells should all be considered when explaining and extrapolating toxicity test results among particles and among species.