Palladium-doped and undoped polystyrene nanoplastics in a chronic toxicity test for higher plants: Impact on soil, plants and ammonium oxidizing bacteria.

There is a lack of knowledge about the fate and impact of microplastics (MPs) and nanoplastics (NPs), as well as their potential uptake and impact on plants and microorganisms. The predicted environmental concentrations (PEC) of frequent polymers in soils are low, and therefore, difficult to detect with the available techniques, which explains the knowledge gaps. Therefore, model particles (polystyrene particles (PS-P), 343 nm) and palladium (Pd) nanoparticle-doped polystyrene particles (PS-Pd-PS-P, 442 nm) were synthesized, characterized, and subsequently applied to agricultural soils (Cambisol, Podzol, PS target contents: 25 mg kg-1, 75 mg kg-1, 225 mg kg-1). A combination of different techniques, such as inductively coupled plasma-mass spectrometry (ICP-MS), pyrolysis-gas chromatography-mass spectrometry (Pyr-GC-MS), dynamic light scattering (DLS), and scanning electron microscopy (SEM), were used to characterize the particles in the dispersions, soils and plants. The spiked soils were applied to a chronical plant toxicity test with oat (Avena sativa). The applied particle contents could be recovered from both soils by ICP-MS (Pd, 89% - 99%), and Pyr-GC-MS (PS, 73% - 120%). Moreover, non-aggregated particles in soils and on oat roots were visualized through SEM. The ratio obtained for the Pd contents in oat roots to that in the Cambisol (2.2-2.7) and the Podzol (2.3-2.6) implied that particles accumulated on the root surface or in the roots. No Pd was detected in the oat shoots, which indicated that no translocation occurred from the roots to the shoots. Despite particle accumulation at or in the roots, no clear effects on plant growth were observed. Furthermore, the soil microorganisms (Podzol) and the soil water repellency (Cambisol, Podzol) showed no clear monotone concentration-response relationship after exposure to PS-P and PS-Pd-PS-P. The findings are complex and illustrate the urgent need for further sophisticated experimental studies to elucidate the impacts of NPs on physicochemical soil function, plants, and soil organisms. The model PS-P doped with Pd nanoparticles significantly enhanced the development and validation of methods for investigating MPs and NPs in environmental matrices, highlighting their considerable potential for further studies.

Biotic and Abiotic Interactions in Freshwater Mesocosms Determine Fate and Toxicity of CuO Nanoparticles.

Transformation, dissolution, and sorption of copper oxide nanoparticles (CuO-NP) play an important role in freshwater ecosystems. We present the first mesocosm experiment on the fate of CuO-NP and the dynamics of the zooplankton community over a period of 12 months. Increasingly low (0.08-0.28 mg Cu L-1) and high (0.99-2.99 mg Cu L-1) concentrations of CuO-NP and CuSO4 (0.10-0.34 mg Cu L-1) were tested in a multiple dosing scenario. At the high applied concentration (CuO-NP_H) CuO-NP aggregated and sank onto the sediment layer, where we recovered 63% of Cu applied. For the low concentration (CuO-NP_L) only 41% of applied copper could be recovered in the sediment. In the water column, the percentage of initially applied Cu recovered was on average 3-fold higher for CuO-NP_L than for CuO-NP_H. Zooplankton abundance was substantially compromised in the treatments CuSO4 (p < 0.001) and CuO-NP_L (p < 0.001). Community analysis indicated that Cladocera were most affected (bk = -0.49), followed by Nematocera (bk = -0.32). The abundance of Cladocera over time and of Dixidae in summer was significantly reduced in the treatment CuO-NP_L (p < 0.001; p < 0.05) compared to the Control. Our results indicate a higher potential for negative impacts on the freshwater community when lower concentrations of CuO-NP (<0.1 mg Cu L-1) enter the ecosystem.

Improving Membrane Filtration for Copper Speciation: Optimal Salt Pretreatments of Polyethersulfone Membranes to Prevent Analyte Retention.

Membrane filtration has been increasingly used to separate dissolved metal ions from dispersed particles, commonly using ultrafiltration membranes, for example, polyethersulfone (PES) membranes with a molecular weight cut-off of 3 kDa. The disadvantage of this technique is an undesired retention of ions, resulting from Coulomb interactions with sulfonic acid groups of the membrane. Therefore, such a membrane acts similar to a cation exchanger column. We solved this drawback by a pretreatment of the PES membrane by other cations. Using CuSO4 as a model compound, we compared the effectiveness of five cations using their salt solutions (Ca2+, Mg2+, Fe2+, Ag+, Ba2+) as pretreatment agents and identified the most effective pretreatment component for a high recovery of copper ions. After membrane filtration without pretreatment, only 52 ± 10%, 64 ± 5%, 75 ± 8%, and 89 ± 7% of nominal Cu concentrations were obtained using initial concentrations of 0.2, 0.5, 1.0, and 4.0 mg L-1, respectively. The efficiency of the investigated cations increased in the order Fe < Ag < Mg < Ca < Ba. Furthermore, we analyzed the most efficient concentration of the pretreatment agent. The best performance was achieved using 0.1 mol L-1 CaCl2 which increased copper recovery to slightly below 100%, even at the lowest tested Cu concentration (recovery 93 ± 10% at 0.2 mg L-1). In the environmentally relevant Cu concentration range of 0.2 mg L-1, 0.1 mol L-1 BaCl2 was identified as the most efficient pretreatment (103 ± 11%).

Ecotoxicity and fate of silver nanomaterial in an outdoor lysimeter study after twofold application by sewage sludge.

The increasing use of antibacterial silver nanomaterials (AgNM) in consumer products leads to their release into sewers. High amounts of AgNM become retained in sewage sludge, which causes their accumulation in agricultural soils when sewage sludge is applied as fertilizer. This increase in AgNM arouses concerns about toxicity to soil organisms and transfer within trophic levels. Long-term field studies simulating the sewage sludge pathway to soils are sparse, and the effects of a second sewage sludge application are unknown. In this perennial field lysimeter study, a twofold application of AgNM (NM-300K, 2 + 3 mg AgNM/kg dry matter soil (DMS)) and a onefold application of silver nitrate (AgNO3, 2 mg Ag/kg DMS) by sewage sludge to the uppermost 20 cm of the soil (Cambisol) were applied. The response of microorganisms to the applications was determined by measuring the inhibition of ammonium-oxidizing bacteria (AOB). Silver concentration in soil, leachates, and crops were measured after acid digestion by inductively coupled plasma mass spectrometry (ICP-MS). Almost no vertical Ag translocation to deeper soil layers and negligible Ag release to leachates suggest that soil is a large sink for AgNM and AgNO3. For AgNM, an increase in toxicity to AOB was shown after the second sewage sludge application. The application of AgNO3 resulted in long-term toxicity comparable to the toxicity of AgNM. Low root uptake from both AgNM- and AgNO3-spiked lysimeters to crops indicates their incomplete immobilization, which is why food chain uptake cannot completely be excluded. However, the root-shoot barrier for wheat (9.8 → 0.1 mg/kg) and skin body barrier for sugar beets (1.0 → 0.2 mg/kg) will further reduce the accumulation within trophic levels. Moreover, the applied AgNM concentration was above the predicted environmental concentration, which is why the root uptake might be negligible in agricultural practice.

Potential of the Red Alga Dixoniella grisea for the Production of Additives for Lubricants.

There is an increasing interest in algae-based raw materials for medical, cosmetic or nutraceutical applications. Additionally, the high diversity of physicochemical properties of the different algal metabolites proposes these substances from microalgae as possible additives in the chemical industry. Among the wide range of natural products from red microalgae, research has mainly focused on extracellular polymers for additive use, while this study also considers the cellular components. The aim of the present study is to analytically characterize the extra- and intracellular molecular composition from the red microalga Dixoniella grisea and to evaluate its potential for being used in the tribological industry. D. grisea samples, fractionated into extracellular polymers (EPS), cells and medium, were examined for their molecular composition. This alga produces a highly viscous polymer, mainly composed of polysaccharides and proteins, being secreted into the culture medium. The EPS and biomass significantly differed in their molecular composition, indicating that they might be used for different bio-additive products. We also show that polysaccharides and proteins were the major chemical compounds in EPS, whereas the content of lipids depended on the separation protocol and the resulting product. Still, they did not represent a major group and were thus classified as a potential valuable side-product. Lyophilized algal fractions obtained from D. grisea were found to be not toxic when EPS were not included. Upon implementation of EPS as a commercial product, further assessment on the environmental toxicity to enchytraeids and other soil organisms is required. Our results provide a possible direction for developing a process to gain an environmentally friendly bio-additive for application in the tribological industry based on a biorefinery approach.

Ion compositions in artificial media control the impact of humic acid on colloidal behaviour, dissolution and speciation of CuO-NP.

The toxicity of copper oxide nanoparticles (CuO-NP) strongly depends on their interactions with the surrounding environment, impacting their dissolution and colloidal stability. This behaviour is studied quite extensively for simplified electrolytes, but information on the behaviour of CuO-NP in more complex artificial media are lacking. In our study, we analysed the colloidal behaviour and considered the speciation of CuO-NP in pure water and three artificial media of different complexity which are used in ecotoxicology. Measurements were done over 7 days in the absence and presence of humic acid (HA) as a model organic molecule. In pure water, the addition of HA lowered the zeta potential from +11 to -41 mV, while in all artificial media, it stayed constantly at about -20 mV. The hydrodynamic diameter of CuO-NP remained unaffected by HA in pure water and seawater, while in porewater and especially in freshwater, HA suppressed strong agglomeration. In pure water, HA strongly increased dissolution to the highest observed value (3% of total Cu), while HA reduced dissolution in all artificial media. Speciation calculations revealed that cations from the media competed with Cu from the NP surface for complexing sites of the HA. This competition may have caused the reduced dissolution in the presence of ions. Furthermore, speciation calculations also suggest that ion composition drove agglomeration behaviour rather than ion concentration: agglomeration was high when divalent cations where the major interaction partner and dominant in relative terms. HA may have reduced the relative dominance and thus altered the agglomeration, aligning it in all media. Summarizing, ion composition and the presence of HA strongly drive the dissolution and agglomeration of CuO-NP in artificial media, consequently, analysing complexation can help to predict environmental behaviour and toxicity.

A new test system for unraveling the effects of soil components on the uptake and toxicity of silver nanoparticles (NM-300K) in simulated pore water.

Fate, bioavailability and toxicity of silver nanoparticles (AgNP) are largely affected by soil properties. Here we focused on how these processes are connected in simulated soil pore water. OECD soil components (sand, kaolin clay, peat) were covered with NM-300K-, AgNO3- and NM-300K dispersant-contaminated water, and Folsomia candida were exposed on the water surface. After 14 days the majority of AgNP was in nano form in sand pore water where also silver uptake was highest. Multilayered cross sections from X-ray micrographs of Collembola exposed to AgNP showed that silver was located in animal areas of direct contact to the contaminated pore water and was ingested. In contrast, in simulated peat pore water only a small fraction of silver was bioavailable. AgNO3 was only bioavailable at the start of the test and not anymore at test end. AgNP and AgNO3 caused immobilization in sand and kaolin pore water while no toxicity was found with peat and OECD soil. A strong correlation (correlation coefficient = 0.901) existed between the concentration of nano silver and immobilization; for ionic silver this was not the case. The dispersant of AgNP was toxic on its own in sand and kaolin pore water. As there are analytical limitations of quantifying AgNP in complex matrices this test system enables a mechanistic view of exposure and uptake of AgNP (and other substances) by F. candida from soil pore water.

Safe-by-Design CuO Nanoparticles via Fe-Doping, Cu-O Bond Length Variation, and Biological Assessment in Cells and Zebrafish Embryos.

The safe implementation of nanotechnology requires nanomaterial hazard assessment in accordance with the material physicochemical properties that trigger the injury response at the nano/bio interface. Since CuO nanoparticles (NPs) are widely used industrially and their dissolution properties play a major role in hazard potential, we hypothesized that tighter bonding of Cu to Fe by particle doping could constitute a safer-by-design approach through decreased dissolution. Accordingly, we designed a combinatorial library in which CuO was doped with 1-10% Fe in a flame spray pyrolysis reactor. The morphology and structural properties were determined by XRD, BET, Raman spectroscopy, HRTEM, EFTEM, and EELS, which demonstrated a significant reduction in the apical Cu-O bond length while simultaneously increasing the planar bond length (Jahn-Teller distortion). Hazard screening was performed in tissue culture cell lines and zebrafish embryos to discern the change in the hazardous effects of doped vs nondoped particles. This demonstrated that with increased levels of doping there was a progressive decrease in cytotoxicity in BEAS-2B and THP-1 cells, as well as an incremental decrease in the rate of hatching interference in zebrafish embryos. The dissolution profiles were determined and the surface reactions taking place in Holtfreter's solution were validated using cyclic voltammetry measurements to demonstrate that the Cu+/Cu2+ and Fe2+/Fe3+ redox species play a major role in the dissolution process of pure and Fe-doped CuO. Altogether, a safe-by-design strategy was implemented for the toxic CuO particles via Fe doping and has been demonstrated for their safe use in the environment.

Influences of use activities and waste management on environmental releases of engineered nanomaterials.

Engineered nanomaterials (ENM) offer enhanced or new functionalities and properties that are used in various products. This also entails potential environmental risks in terms of hazard and exposure. However, hazard and exposure assessment for ENM still suffer from insufficient knowledge particularly for product-related releases and environmental fate and behavior. This study therefore analyzes the multiple impacts of the product use, the properties of the matrix material, and the related waste management system (WMS) on the predicted environmental concentration (PEC) by applying nine prospective life cycle release scenarios based on reasonable assumptions. The products studied here are clothing textiles treated with silver nanoparticles (AgNPs), since they constitute a controversial application. Surprisingly, the results show counter-intuitive increases by a factor of 2.6 in PEC values for the air compartment in minimal AgNP release scenarios. Also, air releases can shift from washing to wearing activity; their associated release points may shift accordingly, potentially altering release hot spots. Additionally, at end-of-life, the fraction of AgNP-residues contained on exported textiles can be increased by 350% when assuming short product lifespans and globalized WMS. It becomes evident that certain combinations of use activities, matrix material characteristics, and WMS can influence the regional PEC by several orders of magnitude. Thus, in the light of the findings and expected ENM market potential, future assessments should consider these aspects to derive precautionary design alternatives and to enable prospective global and regional risk assessments.

Does the exposure mode to ENPs influence their toxicity to aquatic species? A case study with TiO2 nanoparticles and Daphnia magna.

Recent studies suggest that the ecotoxicity of engineered nanoparticles (ENPs) is dependent upon the treatment of ENPs in suspensions (e.g. sonication or use of solvents) and on the mode of exposure to test organisms. We conducted several bioassays with Daphnia magna in order to determine how adverse effects of TiO2 nanoparticles (n-TiO2) are influenced by experimental set-up. Several treatments were applied, including three test media, several treatments of n-TiO2 suspensions (stirring, sonication) and different exposure modes (exposure duration and volume of test suspension). No adverse effects were observed when D. magna were exposed to 50 mL of suspension, regardless of TiO2 concentration (up to 250 mg/L) and exposure duration. Conversely, adverse effects were observed when D. magna were exposed to 2 mL of suspension for 96 h with a 50 % effect concentration EC50 values ranging from 32 mg/L to 82 mg/L. Test media had no significant influence on the outcome of all treatments. For a better mechanistic understanding of the experimental set-up at which adverse effects were observed, the particle size of n-TiO2 in the test media was characterized throughout the test duration. These measurements revealed a fast and strong agglomeration with a secondary particle size in the order of magnitude of micrometers. Our study describes how the effects of n-TiO2 on D .magna are influenced by the duration of exposure and volume of media, highlighting the need for standardization of experimental methods.

Silver nanoparticle-doped zirconia capillaries for enhanced bacterial filtration.

Membrane clogging and biofilm formation are the most serious problems during water filtration. Silver nanoparticle (Agnano) coatings on filtration membranes can prevent bacterial adhesion and the initiation of biofilm formation. In this study, Agnano are immobilized via direct reduction on porous zirconia capillary membranes to generate a nanocomposite material combining the advantages of ceramics being chemically, thermally and mechanically stable with nanosilver, an efficient broadband bactericide for water decontamination. The filtration of bacterial suspensions of the fecal contaminant Escherichia coli reveals highly efficient bacterial retention capacities of the capillaries of 8 log reduction values, fulfilling the requirements on safe drinking water according to the U.S. Environmental Protection Agency. Maximum bacterial loading capacities of the capillary membranes are determined to be 3×10(9)bacterialcells/750mm(2) capillary surface until back flushing is recommendable. The immobilized Agnano remain accessible and exhibit strong bactericidal properties by killing retained bacteria up to maximum bacterial loads of 6×10(8)bacterialcells/750mm(2) capillary surface and the regenerated membranes regain filtration efficiencies of 95-100%. Silver release is moderate as only 0.8% of the initial silver loading is leached during a three-day filtration experiment leading to average silver contaminant levels of 100µg/L.

The coating makes the difference: acute effects of iron oxide nanoparticles on Daphnia magna.

The surface of nanoparticles (NP) is often functionalized with a capping agent to increase their colloidal stability. Having a strong effect on the characteristics of NP, the coating might already determine the risk from NP to organisms and the environment. In this study identical iron oxide nanoparticles (IONP; Ø 5-6nm) were functionalized with four different coatings: ascorbate (ASC-IONP), citrate (CIT-IONP), dextran (DEX-IONP), and polyvinylpyrrolidone (PVP-IONP). Ascorbate and citrate stabilize NP via electrostatic repulsion whereas dextran and polyvinylpyrrolidone are steric stabilizers. All IONP were colloidally stable over several weeks. Their acute effects on neonates of the waterflea Daphnia magna were investigated over 96h. The highest immobilizing effect was found for ASC- and DEX-IONP. In the presence of neonates, both agglomerated or flocculated and adsorbed to the carapace and filtering apparatuses, inducing high immobilization. Lower immobilization was found for CIT-IONP. Their effect was hypothesized to partly originate from an increased release of dissolved iron and the ability to form reactive oxygen species (ROS). Furthermore, incomplete ecdysis occurred at high concentrations of ASC-, DEX-, and CIT-IONP. PVP-IONP did not induce any negative effect, although high quantities were visibly ingested by the daphnids. PVP-IONP had the highest colloidal stability without any occurring agglomeration, adsorption, or dissolution. Only strong swelling of the PVP coating was observed in medium, highly increasing the hydrodynamic diameter. Each coating caused individual effects. Toxicity cannot be correlated to hydrodynamic diameter or the kind of stabilizing forces. Effects are rather linked to decreasing colloidal stability, the release of ions from the core material or the ability to form ROS, respectively.

A miniaturized solid contact test with Arthrobacter globiformis for the assessment of the environmental impact of silver nanoparticles.

Silver nanoparticles (AgNPs) are widely applied for their antibacterial activity. Their increasing use in consumer products implies that they will find their way into the environment via wastewater-treatment plants. The aim of the present study was to compare the ecotoxicological impact of 2 differently designed AgNPs using the solid contact test for the bacterial strain Arthrobacter globiformis. In addition, a miniaturized version of this test system was established, which requires only small-sized samples because AgNPs are produced in small quantities during the design level. The results demonstrate that the solid contact test can be performed in 24-well microplates and that the miniaturized test system fulfills the validity criterion. Soils spiked with AgNPs showed a concentration-dependent reduction of Arthrobacter dehydrogenase activity for both AgNPs and Ag ions (Ag(+)). The toxic effect of the investigated AgNPs on the bacterial viability differed by 1 order of magnitude and can be related to the release of dissolved Ag(+). The release of dissolved Ag(+) can be attributed to particle size and surface area or to the fact that AgNPs are in either metallic or oxide form. Environ

Adaptation of the Daphnia sp. acute toxicity test: miniaturization and prolongation for the testing of nanomaterials.

Manufacturing of nanomaterials (NMs) is often complex and expensive, and their environmental risks are poorly understood or even unknown. An economization of testing NMs is therefore desirable, which can be achieved by miniaturizing test systems. However, the downsizing of test vessels and volumes can enlarge the surface/volume ratio (SVR) which in turn can affect the bioavailable concentration of adsorbing substances like NMs. The present study focused on the miniaturization of the acute toxicity test with Daphnia magna. The adaptations were verified with three reference substances, the non-adsorbing potassium dichromate (K2Cr2O7) and as potentially highly-adsorbing substances silver nanoparticles (AgNPs) and silver nitrate (AgNO3). The miniaturized test was conducted in 24-well microtiter plates (MT) and simultaneously compared to the OECD standard test (ST). Furthermore, the test duration was prolonged from 48 to 96 h since NMs tend to show effects only after extended exposure. The toxicity of K2Cr2O7 and AgNPs continued to increase within the prolonged test span. The test comparisons with K2Cr2O7 did not reveal any significant differences between ST and MT. AgNO3 toxicity was significantly decreased in MT compared to ST due to the enlarged SVR. The toxicity of AgNPs in MT after 24 h was equal to ST. Contrary to our expectations an exposure longer than 24 h resulted in an increase of AgNP toxicity in MT, possibly due to enhanced dissolution of silver. Microtiter plates are appropriate alternative test vessels for the Daphnia sp. acute toxicity test; thus, its miniaturization is feasible. The enlarged SVR has to be taken into account since it can affect the toxicity of potentially adsorbing substances. Furthermore, the standard test duration of 48 h might underestimate the toxicity of many substances, especially of NMs.

Intrinsically green iron oxide nanoparticles? From synthesis via (eco-)toxicology to scenario modelling.

Iron oxide nanoparticles (IONP) are currently being studied as green magnet resonance imaging (MRI) contrast agents. They are also used in huge quantities for environmental remediation and water treatment purposes, although very little is known on the consequences of such applications for organisms and ecosystems. In order to address these questions, we synthesised polyvinylpyrrolidone-coated IONP, characterised the particle dispersion in various media and investigated the consequences of an IONP exposure using an array of biochemical and biological assays. Several theoretical approaches complemented the measurements. In aqueous dispersion IONP had an average hydrodynamic diameter of 25 nm and were stable over six days in most test media, which could also be predicted by stability modelling. The particles were tested in concentrations of up to 100 mg Fe per L. The activity of the enzymes glutathione reductase and acetylcholine esterase was not affected, nor were proliferation, morphology or vitality of mammalian OLN-93 cells although exposure of the cells to 100 mg Fe per L increased the cellular iron content substantially. Only at this concentration, acute toxicity tests with the freshwater flea Daphnia magna revealed slightly, yet insignificantly increased mortality. Two fundamentally different bacterial assays, anaerobic activated sludge bacteria inhibition and a modified sediment contact test with Arthrobacter globiformis, both rendered results contrary to the other assays: at the lowest test concentration (1 mg Fe per L), IONP caused a pronounced inhibition whereas higher concentrations were not effective or even stimulating. Preliminary and prospective risk assessment was exemplified by comparing the application of IONP with gadolinium-based nanoparticles as MRI contrast agents. Predicted environmental concentrations were modelled in two different scenarios, showing that IONP could reduce the environmental exposure of toxic Gd-based particles by more than 50%. Application of the Swiss "Precautionary Matrix for Synthetic Nanomaterials" rendered a low precautionary need for using our IONP as MRI agents and a higher one when using them for remediation or water treatment. Since IONP and (considerably more reactive) zerovalent iron nanoparticles are being used in huge quantities for environmental remediation purposes, it has to be ascertained that these particles pose no risk to either human health or to the environment.