Exposure of humic acid-coated goethite colloids to groundwater does not affect their adsorption of metal(loid)s and their impact on Daphnid mobility.

Engineered humic acid-coated goethite (HA-Goe) colloids find increasing application in in situ remediation of metal(loid)-polluted groundwater. Once introduced into the subsurface, the colloids interact with groundwater altering their physicochemical properties. In comparison to freshly synthesized, unreacted HA-Goe colloids, such alterations could reduce the adsorption affinity towards metal(loid)s and also result in altered ecotoxicological effects. In our study, HA-Goe colloids were exposed to two groundwaters (low vs. high concentrations of metal(loid)s) from two metal(loid)-contaminated sites for 87 days. We investigated (i) the course of HA-Goe ecotoxicity (Daphnia magna immobilization tests), (ii) HA-Goe adsorption properties (multi-element solutions containing As, Cu, Zn, Ni and Co), and (iii) changes in the chemical composition as well as in the mineral and aggregate properties of HA-Goe. The adsorption affinity of HA-Goe decreased in the order As ≈ Cu ≫ Zn > Ni ≈ Co. The metal(loid) adsorption occurred rapidly after mixing prior to the first sampling, while the duration of ongoing exposition to groundwater had no effect on the adsorption of these metal(loid)s. We neither observed a desorption of humic acids from the goethite surface nor alterations in the mineralogy, crystallinity, and surface properties of HA-Goe. Standardized Daphnia magna immobilization tests showed an increased number of mobile organisms with increasing exposure time of HA-Goe to both groundwaters. The decrease in HA-Goe-mediated immobilization of D. magna was strongest within the first 30 d. We attribute this to a shift to smaller sizes due to the breakdown of large HA-Goe aggregates, particularly within the first 30 d. The breakdown of these µm-sized aggregates may result mainly from the repeated shaking of the HA-Goe suspensions. Our study confirms within this particular setting that the tested HA-Goe colloids are suitable for the long-term immobilization of metal(loid)s, while lethal effects on D. magna were negligible.

Field-scale demonstration of in situ immobilization of heavy metals by injecting iron oxide nanoparticle adsorption barriers in groundwater.

Remediation of heavy metal-contaminated aquifers is a challenging process because they cannot be degraded by microorganisms. Together with the usually limited effectiveness of technologies applied today for treatment of heavy metal contaminated groundwater, this creates a need for new remediation technologies. We therefore developed a new treatment, in which permeable adsorption barriers are established in situ in aquifers by the injection of colloidal iron oxides. These adsorption barriers aim at the immobilization of heavy metals in aquifers groundwater, which was assessed in a large-scale field study in a brownfield site. Colloidal iron oxide (goethite) nanoparticles were used to install an in situ adsorption barrier in a very heterogeneous, contaminated aquifer of a brownfield in Asturias, Spain. The groundwater contained high concentrations of heavy metals with up to 25 mg/L zinc, 1.3 mg/L lead, 40 mg/L copper, 0.1 mg/L nickel and other minor heavy metal pollutants below 1 mg/L. High amounts of zinc (>900 mg/kg), lead (>2000 mg/kg), nickel (>190 mg/kg) were also present in the sediment. Ca. 1500 kg of goethite nanoparticles of 461 ± 266 nm diameter were injected at low pressure (< 0.6 bar) into the aquifer through nine screened injection wells. For each injection well, a radius of influence of at least 2.5 m was achieved within 8 h, creating an in situ barrier of 22 × 3 × 9 m. Despite the extremely high heavy metal contamination and the strong heterogeneity of the aquifer, successful immobilization of contaminants was observed in the tested area. The contaminant concentrations were strongly reduced immediately after the injection and the abatement of the heavy metals continued for a total post-injection monitoring period of 189 days. The iron oxide particles were found to adsorb heavy metals even at pH-values between 4 and 6, where low adsorption would have been expected. The study demonstrated the applicability of iron oxide nanoparticles for installing adsorption barriers for containment of heavy metals in contaminated groundwater under real conditions.

Earthworm Uptake Routes and Rates of Ionic Zn and ZnO Nanoparticles at Realistic Concentrations, Traced Using Stable Isotope Labeling.

The environmental behavior of ZnO nanoparticles (NPs), their availability to, uptake pathways by, and biokinetics in the earthworm Lumbricus rubellus were investigated using stable isotope labeling. Zinc isotopically enriched to 99.5% in (68)Zn ((68)Zn-E) was used to prepare (68)ZnO NPs and a dissolved phase of (68)Zn for comparison. These materials enabled tracing of environmentally relevant (below background) NP additions to soil of only 5 mg (68)Zn-E kg(-1). Uptake routes were isolated by introducing earthworms with sealed and unsealed mouthparts into test soils for up to 72 h. The Zn isotope compositions of the soils, pore waters and earthworms were then determined using multiple collector inductively coupled plasma mass spectrometry. Detection and quantification of (68)Zn-E in earthworm tissue was possible after only 4 h of dermal exposure, when the uptake of (68)Zn-E had increased the total Zn tissue concentration by 0.03‰. The results demonstrate that at these realistic exposure concentrations there is no distinguishable difference between the uptake of the two forms of Zn by the earthworm L. rubellus, with the dietary pathway accounting for ∼95% of total uptake. This stands in contrast to comparable studies where high dosing levels were used and dermal uptake is dominant.

Different routes, same pathways: Molecular mechanisms under silver ion and nanoparticle exposures in the soil sentinel Eisenia fetida.

Use of nanotechnology products is increasing; with silver (Ag) nanoparticles particularly widely used. A key uncertainty surrounding the risk assessment of AgNPs is whether their effects are driven through the same mechanism of action that underlies the toxic effects of Ag ions. We present the first full transcriptome study of the effects of Ag ions and NPs in an ecotoxicological model soil invertebrate, the earthworm Eisenia fetida. Gene expression analyses indicated similar mechanisms for both silver forms with toxicity being exerted through pathways related to ribosome function, sugar and protein metabolism, molecular stress, disruption of energy production and histones. The main difference seen between Ag ions and NPs was associated with potential toxicokinetic effects related to cellular internalisation and communication, with pathways related to endocytosis and cilia being significantly enriched. These results point to a common final toxicodynamic response, but initial internalisation driven by different exposure routes and toxicokinetic mechanisms.

Uptake routes and toxicokinetics of silver nanoparticles and silver ions in the earthworm Lumbricus rubellus.

Current bioavailability models, such as the free ion activity model and biotic ligand model, explicitly consider that metal exposure will be mainly to the dissolved metal in ionic form. With the rise of nanotechnology products and the increasing release of metal-based nanoparticles (NPs) to the environment, such models may increasingly be applied to support risk assessment. It is not immediately clear, however, whether the assumption of metal ion exposure will be relevant for NPs. Using an established approach of oral gluing, a toxicokinetics study was conducted to investigate the routes of silver nanoparticles (AgNPs) and Ag(+) ion uptake in the soil-dwelling earthworm Lumbricus rubellus. The results indicated that a significant part of the Ag uptake in the earthworms is through oral/gut uptake for both Ag(+) ions and NPs. Thus, sealing the mouth reduced Ag uptake by between 40% and 75%. An X-ray analysis of the internal distribution of Ag in transverse sections confirmed the presence of increased Ag concentrations in exposed earthworm tissues. For the AgNPs but not the Ag(+) ions, high concentrations were associated with the gut wall, liver-like chloragogenous tissue, and nephridia, which suggest a pathway for AgNP uptake, detoxification, and excretion via these organs. Overall, the results indicate that Ag in the ionic and NP forms is assimilated and internally distributed in earthworms and that this uptake occurs predominantly via the gut epithelium and less so via the body wall. The importance of oral exposure questions the application of current metal bioavailability models, which implicitly consider that the dominant route of exposure is via the soil solution, for bioavailability assessment and modeling of metal-based NPs.

Short-term soil bioassays may not reveal the full toxicity potential for nanomaterials; bioavailability and toxicity of silver ions (AgNO3) and silver nanoparticles to earthworm Eisenia fetida in long-term aged soils.

This study investigated if standard risk assessment hazard tests are long enough to adequately provide the worst case exposure for nanomaterials. This study therefore determined the comparative effects of the aging on the bioavailability and toxicity to earthworms of soils dosed with silver ions and silver nanoparticles (Ag NP) for 1, 9, 30 & 52 weeks, and related this to the total Ag in the soil, Ag in soil pore water and earthworm tissue Ag concentrations. For ionic Ag, a classical pattern of reduced bioavailability and toxicity with time aged in the soil was observed. For the Ag NP, toxicity increased with time apparently driven by Ag ion dissolution from the added Ag NPs. Internal Ag in the earthworms did not always explain toxicity and suggested the presence of an internalised, low-toxicity Ag fraction (as intact or transformed NPs) after shorter aging times. Our results indicate that short-term exposures, without long-term soil aging, are not able to properly assess the environmental risk of Ag NPs and that ultimately, with aging time, Ag ion and Ag NP effect will merge to a common value.

Soil pH effects on the comparative toxicity of dissolved zinc, non-nano and nano ZnO to the earthworm Eisenia fetida.

To determine how soil properties influence nanoparticle (NP) fate, bioavailability and toxicity, this study compared the toxicity of nano zinc oxide (ZnO NPs), non-nano ZnO and ionic ZnCl2 to the earthworm Eisenia fetida in a natural soil at three pH levels. NP characterisation indicated that reaction with the soil media greatly controls ZnO properties. Three main conclusions were drawn. First that Zn toxicity, especially for reproduction, was influenced by pH for all Zn forms. This can be linked to the influence of pH on Zn dissolution. Secondly, that ZnO fate, toxicity and bioaccumulation were similar (including relationships with pH) for both ZnO forms, indicating the absence of NP-specific effects. Finally, earthworm Zn concentrations were higher in worms exposed to ZnO compared to ZnCl2, despite the greater toxicity of the ionic form. This observation suggests the importance of considering the relationship between uptake and toxicity in nanotoxicology studies.

Assessing the impact of organic and inorganic amendments on the toxicity and bioavailability of a metal-contaminated soil to the earthworm Eisenia andrei.

Metal-contaminated soil, from the El Arteal mining district (SE Spain), was remediated with organic (6% compost) and inorganic amendments (8% marble sludge) to reduce the mobility of metals and to modify its potential environmental impact. Different measures of metal bioavailability (chemical analysis; survival, growth, reproduction and bioaccumulation in the earthworm Eisenia andrei), were tested in order to evaluate the efficacy of organic and inorganic amendments as immobilizing agents in reducing metal (bio)availability in the contaminated soil. The inorganic amendment reduced water and CaCl2-extractable concentrations of Cd, Pb, and Zn, while the organic amendment increased these concentrations compared to the untreated soil. The inorganic treatment did not significantly reduce toxicity for the earthworm E. andrei after 28 days exposure. The organic amendment however, made the metal-contaminated soil more toxic to the earthworms, with all earthworms dying in undiluted soil and completely inhibiting reproduction at concentrations higher than 25%. This may be due to increased available metal concentrations and higher electrical conductivity in the compost-amended soil. No effects of organic and inorganic treatments on metal bioaccumulation in the earthworms were found and metal concentrations in the earthworms increased with increasing total soil concentrations.

Sorption, dissolution and pH determine the long-term equilibration and toxicity of coated and uncoated ZnO nanoparticles in soil.

To assess the effect of long-term dissolution on bioavailability and toxicity, triethoxyoctylsilane coated and uncoated zinc oxide nanoparticles (ZnO-NP), non-nano ZnO and ZnCl2 were equilibrated in natural soil for up to twelve months. Zn concentrations in pore water increased with time for all ZnO forms but peaked at intermediate concentrations of ZnO-NP and non-nano ZnO, while for coated ZnO-NP such a clear peak only was seen after 12 months. Dose-related increases in soil pH may explain decreased soluble Zn levels due to fixation of Zn released from ZnO at higher soil concentrations. At T = 0 uncoated ZnO-NP and non-nano ZnO were equally toxic to the springtail Folsomia candida, but not as toxic as coated ZnO-NP, and ZnCl2 being most toxic. After three months equilibration toxicity to F. candida was already reduced for all Zn forms, except for coated ZnO-NP which showed reduced toxicity only after 12 months equilibration.

Effect of different spiking procedures on the distribution and toxicity of ZnO nanoparticles in soil.

Due to the difficulty in dispersing some engineered nanomaterials in exposure media, realizing homogeneous distributions of nanoparticles (NP) in soil may pose major challenges. The present study investigated the distribution of zinc oxide (ZnO) NP (30 nm) and non-nano ZnO (200 nm) in natural soil using two different spiking procedures, i.e. as dry powder and as suspension in soil extract. Both spiking procedures showed a good recovery (>85 %) of zinc and based on total zinc concentrations no difference was found between the two spiking methods. Both spiking procedures resulted in a fairly homogeneous distribution of the ZnO particles in soil, as evidenced by the low variation in total zinc concentration between replicate samples (<12 % in most cases). Survival of Folsomia candida in soil spiked at concentrations up to 6,400 mg Zn kg(-1) d.w. was not affected for both compounds. Reproduction was reduced in a concentration-dependent manner with EC50 values of 3,159 and 2,914 mg Zn kg(-1) d.w. for 30 and 200 nm ZnO spiked as dry powder and 3,593 and 5,633 mg Zn kg(-1) d.w. introduced as suspension. Toxicity of ZnO at 30 and 200 nm did not differ. We conclude that the ZnO particle toxicity is not size related and that the spiking of the soil with ZnO as dry powder or as a suspension in soil extract does not affect its toxicity to F. candida.

Influence of soil properties on molybdenum uptake and elimination kinetics in the earthworm Eisenia andrei.

This study aimed at determining the uptake and elimination kinetics of molybdenum in the earthworm Eisenia andrei, and the influence of soil properties on molybdenum bioaccumulation. Three natural and four artificial soils were spiked at concentrations of 10 and 100 microg Mo g(-1) dry soil. Earthworms were exposed individually to spiked soils and sampled at different time intervals for 21 d. Remaining earthworms were transferred to non-spiked soil to determine elimination, also for 21 d. Uptake and elimination rate constants and bioaccumulation factors (BAFs) were calculated using a one-compartment model, and related to total, water and CaCl2 extractable molybdenum concentrations in soil. Molybdenum was rapidly accumulated by the earthworms with uptake rate constants between 0.05 and 1.70 g dry soil g(-1) dry earthworm d(-1), and equilibrium generally was reached within 10 d. Molybdenum was not strongly bioaccumulated (BAF