Incubation media modify silver nanoparticle toxicity for whitefish (Coregonus lavaretus) and roach (Rutilus rutilus) embryos.

Toxicological studies were performed to examine silver nanoparticle (AgNP, size: 14.4 ± 2.5 nm) transformation within three different test media and consequent effects on embryos of whitefish (Coregonus lavaretus) and roach (Rutilus rutilus). The test media, namely ASTM very hard water, ISO standard dilution medium, and natural lake water differed predominantly in ionic strength. Total silver was determined using inductively coupled plasma mass spectrometry (ICP-MS), while AgNPs were characterized by transmission electron microscopy and single particle ICP-MS. Silver species distributions were estimated via thermodynamic speciation calculations. Data demonstrated that increased AgNP dissolution accompanied by decreasing ionic strength of the test medium did not occur as noted in other studies. Further, other physicochemical parameters including AgNP size and metallic species distribution did not markedly affect AgNP-induced toxicity. Irrespective of the test medium, C. lavaretus were more sensitive to AgNP exposure (median lethal concentration after 8 weeks: 0.51-0.73 mg/L) compared to R. rutilus, where adverse effects were only observed at 5 mg/L in natural lake water. In addition, AgNP-induced toxicity was lower in the two standard test media compared to natural lake water. Currently, there are no apparent studies assessing simultaneously the sensitivity of C. lavaretus and R. rutilus to AgNP exposure. Therefore, the aim of this study was to (1) investigate AgNP-induced toxicity in C. lavaretus and R. rutilus cohabiting in the same aquatic environment and (2) the role played by test media in the observed effects of AgNPs on these aquatic species.

Effects of copper in Daphnia are modulated by nanosized titanium dioxide and natural organic matter: what is the impact of aging duration?

During its aquatic life cycle, nanosized titanium dioxide (nTiO2) may interact with natural organic matter (NOM) ultimately altering the ecotoxicity of co-occurring chemical stressors such as heavy metals (e.g. copper (Cu)). In this context, the following hypotheses were tested: (1) aging of nTiO2 along with Cu reduces Cu toxicity, (2) nTiO2 agglomerates have a lower potential to reduce Cu toxicity and (3) aging of nTiO2 in presence of NOM reduces Cu toxicity further. A multifactorial test design crossing three nTiO2 levels (0.0, 0.6 and 3.0 mg/L) with two levels of NOM (0 versus 8 mg total organic carbon (TOC)/L) and seven nominal Cu concentrations (ranging from 0 to 1536 µg/L) aged in ASTM medium for 0, 1, 3 and 6 days was realised, while two aging scenarios were applied (type 1: nTiO2 jointly aged with Cu; type 2: Cu added after nTiO2 aging). Subsequently, Cu toxicity was assessed using the immobility of Daphnia magna after 48 h of exposure as response variable. The experiments revealed that neither aging duration nor the extent of agglomeration (type 1 vs. type 2 aging) has a substantial impact on Cu induced toxicity. Moreover, it was confirmed that the presence of NOM substantially reduced Cu induced toxicity, independent of the aging scenario and duration. More importantly, the data suggest the ingestion of Cu loaded nTiO2 as additional exposure pathway contributing to Cu toxicity. In conclusion, it seems unlikely that nTiO2 concentrations currently detected in or predicted for aquatic ecosystems, which are at least one order of magnitude below the concentration tested here, influence Cu toxicity meaningfully.

Morphology, structure, and composition of sulfidized silver nanoparticles and their aggregation dynamics in river water.

The sulfidized form represents an environmentally relevant transformation state of silver nanoparticles (Ag-NPs) released into natural systems via wastewater route. However, the detailed characterization of sulfidized silver nanoparticles (S-Ag-NPs) is missing and their colloidal stability in aquatic systems is only insufficiently studied. The aim of this study was to systematically evaluate the surface properties, morphology, structure, composition, as well as aggregation dynamics of S-Ag-NPs in synthetic and natural river water. The S-Ag-NPs were prepared by sulfidation of citrate-coated silver nanoparticles (Cit-Ag-NPs). The sulfidation of Ag-NPs was accompanied by the formation of fiber-like Ag2S nano-bridges, Ag0-Ag2S core-shell structures, and hollow regions. In contrast to the published literature, the nano-bridges were thinner (2-9 nm) and longer (up to 60 nm), they formed at higher S2-/Ag molar ratio (2.041), and the formation of the core-shell structures was observed even in the absence of natural organic matter (NOM). Furthermore, we observed selective sulfidation of nanoparticles which can induce the hot spots for the release of toxic Ag+ ions. The critical coagulation concentration (CCC) of Ca2+ determined for S-Ag-NPs in reconstituted river water was 2.47 ± 0.23 mmol/L and thus higher than the CCC obtained for Cit-Ag-NPs in our earlier study revealing higher colloidal stability of S-Ag-NPs. In natural river water, S-Ag-NPs were also colloidally more stable compared to the Cit-Ag-NPs. Furthermore, the stabilizing effect of NOM was much higher for S-Ag-NPs than for Cit-Ag-NPs. For S-Ag-NPs stabilized by a low amount of citrate, we expect longer residence times in the water phase of rivers and thus higher risk for aquatic organisms. In contrast to this, the pristine Cit-Ag-NPs are expected to be accumulated faster in the sediments representing higher risk for benthic organisms. This study contributes to better understanding of environmental fate and effects of Ag-NPs released via wastewater route.

The fate of silver nanoparticles in riverbank filtration systems - The role of biological components and flow velocity.

Riverbank filtration is a natural process that may ensure the cleaning of surface water for producing drinking water. For silver nanoparticles (AgNP), physico-chemical interaction with sediment surfaces is one major retention mechanism. However, the effect of flow velocity and the importance of biological retention, such as AgNP attachment to biomass, are not well understood, yet. We investigated AgNP (c = 0.6 mg L-1) transport at different spatial and temporal scales in pristine and previously pond water-aged sediment columns. Transport of AgNP under near-natural conditions was studied in a long-term riverbank filtration experiment over the course of one month with changing flow scenarios (i.e. transport at 0.7 m d-1, stagnation, and remobilization at 1.7 m d-1). To elucidate retention processes, we conducted small-scale lab column experiments at low (0.2 m d-1) and high (0.7 m d-1) flow rate using pristine and aged sediments. Overall, AgNP accumulated in the upper centimeters of the sediment both in lab and outdoor experiments. In the lab study, retention of AgNP by attachment to biological components was very effective under high and low flow rate with nearly complete NP accumulation in the upper 2 mm. When organic material was absent, abiotic filtration mechanisms led to NP retention in the upper 5 to 7 cm of the column. In the long-term study, AgNP were transported up to a depth of 25 cm. For the pristine sediment in the lab study and the outdoor experiments only erratic particle breakthrough was detected in a depth of 15 cm. We conclude that physico-chemical interactions of AgNP with sediment surfaces are efficient in retaining AgNP. The presence of organic material provides additional retention sites which increase the filtration capacity of the system. Nevertheless, erratic breakthrough events might transport NP into deeper sediment layers.

Exposure pathway dependent effects of titanium dioxide and silver nanoparticles on the benthic amphipod Gammarus fossarum.

The increasing production of engineered inorganic nanoparticles (EINPs) elevates their release into aquatic ecosystems raising concerns about associated environmental risks. Numerous investigations indicate sediments as the final sink, facilitating the exposure of benthic species to EINPs. Although reports of sub-lethal EINP effects on benthic species are increasing, the importance of exposure pathways (either waterborne or dietary) is poorly understood. This study investigates the influence of two EINPs, namely titanium dioxide (nTiO2) and silver (nAg), on the benthic model organism Gammarus fossarum specifically addressing the relative relevance of these pathways. For each type of EINP an individual 30-day long bioassay was conducted, applying a two-factorial test design. The factors include the presence or absence of the EINPs (nTiO2: ∼80 nm, 4 mg/L or nAg: ∼30 nm, 0.125 mg/L; n = 30) in the water phase (waterborne), combined with a preceding 6-day long aging of their diet (black alder leaves) also in presence or absence of the EINPs (dietary). Response variables were mortality, food consumption, feces production and energy assimilation. Additionally, the physiological fitness was examined using lipid content and dry weight of the organisms as measures. Results revealed a significantly reduced energy assimilation (up to ∼30%) in G. fossarum induced by waterborne exposure towards nTiO2. In contrast, the dietary exposure towards nAg significantly increased the organisms' energy assimilation (up to ∼50%). Hence, exposure pathway dependent effects of EINPs cannot be generalized and remain particle specific resting upon their intrinsic properties affecting their potential to interact with the surrounding environment. As a result of the different properties of the EINPs used in this study, we clearly demonstrated variations in type and direction of observed effects in G. fossarum. The results of the present study are thus supporting current approaches for nano-specific grouping that might enable an enhanced accuracy in predicting EINP effects facilitating their environmental risk assessment.

Retention and remobilization mechanisms of environmentally aged silver nanoparticles in an artificial riverbank filtration system.

Riverbank filtration systems are important structures that ensure the cleaning of infiltrating surface water for drinking water production. In our study, we investigated the potential risk for a breakthrough of environmentally aged silver nanoparticles (Ag NP) through these systems. Additionally, we identified factors leading to the remobilization of Ag NP accumulated in surficial sediment layers in order to gain insights into remobilization mechanisms. We conducted column experiments with Ag NP in an outdoor pilot plant consisting of water-saturated sediment columns mimicking a riverbank filtration system. The NP had previously been aged in river water, soil extract, and ultrapure water, respectively. We investigated the depth-dependent breakthrough and retention of NP. In subsequent batch experiments, we studied the processes responsible for a remobilization of Ag NP retained in the upper 10 cm of the sediments, induced by ionic strength reduction, natural organic matter (NOM), and mechanical forces. We determined the amount of remobilized Ag by ICP-MS and differentiated between particulate and ionic Ag after remobilization using GFAAS. The presence of Ag-containing heteroaggregates was investigated by combining filtration with single-particle ICP-MS. Single and erratic Ag breakthrough events were mainly found in 30 cm depth and Ag NP were accumulated in the upper 20 cm of the columns. Soil-aged Ag NP showed the lowest retention of only 54%. Remobilization was induced by the reduction of ionic strength and the presence of NOM in combination with mechanical forces. The presence of calcium in the aging- as well as the remobilizing media reduced the remobilization potential. Silver NP were mainly remobilized as heteroaggregates with natural colloids, while dissolution played a minor role. Our study indicates that the breakthrough potential of Ag NP in riverbank filtration systems is generally low, but the aging in soil increases their mobility. Remobilization processes are associated to co-mobilization with natural colloids.

Effects of low dose silver nanoparticle treatment on the structure and community composition of bacterial freshwater biofilms.

The application of engineered silver nanoparticles (AgNPs) in a considerable amount of registered commercial products inevitably will result in the continuous release of AgNPs into the natural aquatic environment. Therefore, native biofilms, as the prominent life form of microorganisms in almost all known ecosystems, will be subjected to AgNP exposure. Despite the exponentially growing research activities worldwide, it is still difficult to assess nanoparticle-mediated toxicity in natural environments. In order to obtain an ecotoxicologically relevant exposure scenario, we performed experiments with artificial stream mesocosm systems approaching low dose AgNP concentrations close to predicted environmental concentrations. Pregrown freshwater biofilms were exposed for 14 days to citrate-stabilized AgNPs at a concentration of 600 µg l-1 in two commonly used sizes (30 and 70 nm). Sublethal effects of AgNP treatment were assessed with regard to biofilm structure by gravimetric measurements (biofilm thickness and density) and by two biomass parameters, chlorophyll a and protein content. The composition of bacterial biofilm communities was characterized by t-RFLP fingerprinting combined with phylogenetic studies based on the 16S gene. After 14 days of treatment, the structural parameters of the biofilm such as thickness, density, and chlorophyll a and protein content were not statistically significantly changed by AgNP exposure. Furthermore, t-RFLP fingerprint analysis showed that the bacterial diversity was not diminished by AgNPs, as calculated by Shannon Wiener and evenness indices. Nevertheless, t-RFLP analysis also indicated that AgNPs led to an altered biofilm community composition as was shown by cluster analysis and multidimensional scaling (MDS) based on the Bray Curtis index. Sequence analysis of cloned 16S rRNA genes further revealed that changes in community composition were related with the displacement of putatively AgNP-sensitive bacterial taxa Actinobacteria, Chloroflexi, and Cyanobacteria by taxa known for their enhanced adaptability towards metal stress, such as Acidobacteria, Sphingomonadales, and Comamonadaceae. This measurable community shift, even after low dose AgNP treatment, causes serious concerns with respect to the broad application of AgNPs and their potentially adverse impact on the ecological function of lotic biofilms, such as biodegradation or biostabilization.

Nanoparticles in the environment: where do we come from, where do we go to?

Nanoparticles serve various industrial and domestic purposes which is reflected in their steadily increasing production volume. This economic success comes along with their presence in the environment and the risk of potentially adverse effects in natural systems. Over the last decade, substantial progress regarding the understanding of sources, fate, and effects of nanoparticles has been made. Predictions of environmental concentrations based on modelling approaches could recently be confirmed by measured concentrations in the field. Nonetheless, analytical techniques are, as covered elsewhere, still under development to more efficiently and reliably characterize and quantify nanoparticles, as well as to detect them in complex environmental matrixes. Simultaneously, the effects of nanoparticles on aquatic and terrestrial systems have received increasing attention. While the debate on the relevance of nanoparticle-released metal ions for their toxicity is still ongoing, it is a re-occurring phenomenon that inert nanoparticles are able to interact with biota through physical pathways such as biological surface coating. This among others interferes with the growth and behaviour of exposed organisms. Moreover, co-occurring contaminants interact with nanoparticles. There is multiple evidence suggesting nanoparticles as a sink for organic and inorganic co-contaminants. On the other hand, in the presence of nanoparticles, repeatedly an elevated effect on the test species induced by the co-contaminants has been reported. In this paper, we highlight recent achievements in the field of nano-ecotoxicology in both aquatic and terrestrial systems but also refer to substantial gaps that require further attention in the future.

Implications of Pony Lake Fulvic Acid for the Aggregation and Dissolution of Oppositely Charged Surface-Coated Silver Nanoparticles and Their Ecotoxicological Effects on Daphnia magna.

Citrate (Cit) and polyethylenimine (BPEI)-coated silver nanoparticles (AgNPs) were used to understand how the type of capping agents and surface charge affect their colloidal stability, dissolution, and ecotoxicity in the absence/presence of Pony Lake Fulvic Acid (PLFA). In the presence of PLFA, Cit-AgNPs were stabilized, while BPEI-AgNPs were aggregated. The aggregation of BPEI-AgNPs decreased with the time, and their stabilizing effect increased at high PLFA concentration. The dissolution also differed between both AgNPs and was influenced by the PLFA concentration. Generally, BPEI-AgNPs showed a lower amount of dissolved Ag than Cit-AgNPs. The dissolved Ag concentration decreased for both AgNPs at low PLFA concentration (5 mg/L). In contrast, the extent of nanoparticle dissolution increased at high PLFA concentration (30 mg/L) but only for BPEI-AgNPs. In the absence of PLFA, the ecotoxicity of Cit-AgNPs to Daphnia magna was higher than that of BPEI-AgNPs. However, the ecotoxicity of AgNPs in the presence of PLFA was up to 70% lower than in their absence. We demonstrated that the differences in colloidal stability, dissolution, and ecotoxicity may be attributed to the different capping agents, surface charge, and concentration of natural organic matter (NOM) as well as to the formation of dissolved Ag complexes with NOM.

Transport of soil-aged silver nanoparticles in unsaturated sand.

Engineered nanoparticles released into soils may be coated with humic substances, potentially modifying their surface properties. Due to their amphiphilic nature, humic coating is expected to affect interaction of nanoparticle at the air-water interface. In this study, we explored the roles of the air-water interface and solid-water interface as potential sites for nanoparticle attachment and the importance of hydrophobic interactions for nanoparticle attachment at the air-water interface. By exposing Ag nanoparticles to soil solution extracted from the upper soil horizon of a floodplain soil, the mobility of the resulting "soil-aged" Ag nanoparticles was investigated and compared with the mobility of citrate-coated Ag nanoparticles as investigated in an earlier study. The mobility was determined as a function of hydrologic conditions and solution chemistry using column breakthrough curves and numerical modeling. Specifically, we compared the mobility of both types of nanoparticles for different unsaturated flow conditions and for pH=5 and pH=9. The soil-aged Ag NP were less mobile at pH=5 than at pH=9 due to lower electrostatic repulsion at pH=5 for both types of interfaces. Moreover, the physical flow field at different water contents modified the impact of chemical forces at the solid-water interface. An extended Derjaguin-Landau-Verwey-Overbeek (eDLVO) model did not provide satisfactory explanation of the observed transport phenomena unlike for the citrate-coated case. For instance, the eDLVO model assuming sphere-plate geometry predicts a high energy barrier (>90 kT) for the solid-water interface, indicating that nanoparticle attachment is less likely. Furthermore, retardation through reversible sorption at the air-water interface was probably less relevant for soil-aged nanoparticles than for citrate-coated nanoparticles. An additional cation bridging mechanism and straining within the flow field may have enhanced nanoparticle retention at the solid-water interface. The results indicate that the mobility of engineered Ag nanoparticles is sensitive to solution chemistry, especially pH and the concentration of multivalent cations, and to the unsaturated flow conditions influencing particle interaction at biogeochemical interfaces.

Sublethal concentrations of silver nanoparticles affect the mechanical stability of biofilms.

Bacterial biofilms are most likely confronted with silver nanoparticles (Ag NPs) as a pollutant stressor in aquatic systems. In this study, biofilms of Aquabacterium citratiphilum were exposed for 20 h to 30 and 70 nm citrate stabilized Ag NPs in low-dose concentrations ranging from 600 to 2400 µg l-1, and the Ag NP-mediated effects on descriptive, structural, and functional biofilm characteristics, including viability, protein content, architecture, and mechanical stability, were investigated. Viability, based on the bacterial cell membrane integrity of A. citratiphilum, as determined by epifluorescence microscopy, remained unaffected after Ag NP exposure. Moreover, in contrast to information in the current literature, protein contents of cells and extracellular polymeric substances (EPS) and biofilm architecture, including dry mass, thickness, and density, were not significantly impacted by exposure to Ag NPs. However, the biofilms themselves served as effective sinks for Ag NPs, exhibiting enrichment factors from 5 to 8. Biofilms showed a greater capacity to accumulate 30 nm sized Ag NPs than 70 nm Ag NPs. Furthermore, Ag NPs significantly threatened the mechanical stability of biofilms, as determined by a newly developed assay. For 30 nm Ag NPs, the mechanical stability of biofilms decreased as the Ag NP concentrations applied to them increased. In contrast, 70 nm Ag NPs produced a similar decrease in mechanical stability for each applied concentration. Overall, this finding demonstrates that exposure to Ag NPs triggers remarkable changes in biofilm adhesion and/or cohesiveness. Because of biofilm-mediated ecological services, this response raises environmental concerns regarding Ag NP release into freshwater systems, even in sublethal concentrations.

Transport of citrate-coated silver nanoparticles in unsaturated sand.

Chemical factors and physical constraints lead to coupled effects during particle transport in unsaturated porous media. Studies on unsaturated transport as typical for soils are currently scarce. In unsaturated porous media, particle mobility is determined by the existence of an air-water interface in addition to a solid-water interface. To this end, we measured breakthrough curves and retention profiles of citrate-coated Ag nanoparticles in unsaturated sand at two pH values (5 and 9) and three different flow rates corresponding to different water contents with 1 mM KNO3 as background electrolyte. The classical DLVO theory suggests unfavorable deposition conditions at the air-water and solid-water interfaces. The breakthrough curves indicate modification in curve shapes and retardation of nanoparticles compared to inert solute. Retention profiles show sensitivity to flow rate and pH and this ranged from almost no retention for the highest flow rate at pH=9 to almost complete retention for the lowest flow rate at pH=5. Modeling of the breakthrough curves, thus, required coupling two parallel processes: a kinetically controlled attachment process far from equilibrium, responsible for the shape modification, and an equilibrium sorption, responsible for particle retardation. The non-equilibrium process and equilibrium sorption are suggested to relate to the solid-water and air-water interfaces, respectively. This is supported by the DLVO model extended for hydrophobic interactions which suggests reversible attachment, characterized by a secondary minimum (depth 3-5 kT) and a repulsive barrier at the air-water interface. In contrast, the solid-water interface is characterized by a significant repulsive barrier and the absence of a secondary minimum suggesting kinetically controlled and non-equilibrium interaction. This study provides new insights into particle transport in unsaturated porous media and offers a model concept representing the relevant processes.

Understanding the fate and biological effects of Ag- and TiO2-nanoparticles in the environment: The quest for advanced analytics and interdisciplinary concepts.

Engineered inorganic nanoparticles (EINP) from consumers' products and industrial applications, especially silver and titanium dioxide nanoparticles (NP), are emitted into the aquatic and terrestrial environments in increasing amounts. However, the current knowledge on their environmental fate and biological effects is diverse and renders reliable predictions complicated. This review critically evaluates existing knowledge on colloidal aging mechanisms, biological functioning and transport of Ag NP and TiO2 NP in water and soil and it discusses challenges for concepts, experimental approaches and analytical methods in order to obtain a comprehensive understanding of the processes linking NP fate and effects. Ag NP undergo dissolution and oxidation with Ag2S as a thermodynamically determined endpoint. Nonetheless, Ag NP also undergo colloidal transformations in the nanoparticulate state and may act as carriers for other substances. Ag NP and TiO2 NP can have adverse biological effects on organisms. Whereas Ag NP reveal higher colloidal stability and mobility, the efficiency of NOM as a stabilizing agent is greater towards TiO2 NP than towards Ag NP, and multivalent cations can dominate the colloidal behavior over NOM. Many of the past analytical obstacles have been overcome just recently. Single particle ICP-MS based methods in combination with field flow fractionation techniques and hydrodynamic chromatography have the potential to fill the gaps currently hampering a comprehensive understanding of fate and effects also at a low field relevant concentrations. These analytical developments will allow for mechanistically orientated research and transfer to a larger set of EINP. This includes separating processes driven by NP specific properties and bulk chemical properties, categorization of effect-triggering pathways directing the EINP effects towards specific recipients, and identification of dominant environmental parameters triggering fate and effect of EINP in specific ecosystems (e.g. soil, lake, or riverine systems).

Effects of silver nanoparticle properties, media pH and dissolved organic matter on toxicity to Daphnia magna.

Studies assessing the acute and chronic toxicity of silver nanoparticle (nAg) materials rarely consider potential implications of environmental variables. In order to increase our understanding in this respect, we investigated the acute and chronic effects of various nAg materials on Daphnia magna. Thereby, different nanoparticle size classes with a citrate coating (20-, ~30-, 60- as well as 100-nm nAg) and one size class without any coating (140 nm) were tested, considering at the same time two pH levels (6.5 and 8.0) as well as the absence or presence of dissolved organic matter (DOM; <0.1 or 8.0 mg total organic carbon/L). Results display a reduced toxicity of nAg in media with higher pH and the presence of DOM as well as increasing initial particle size, if similarly coated. This suggests that the associated fraction of Ag species <2 nm (including Ag(+)) is driving the nAg toxicity. This hypothesis is supported by normalizing the 48-h EC50-values to Ag species <2 nm, which displays comparable toxicity estimates for the majority of the nAg materials assessed. It may therefore be concluded that a combination of both the particle characteristics, i.e. its initial size and surface coating, and environmental factors trigger the toxicity of ion-releasing nanoparticles.

The fate of silver nanoparticles in soil solution--Sorption of solutes and aggregation.

Nanoparticles enter soils through various pathways. In the soil, they undergo various interactions with the solution and the solid phase. We tested the following hypotheses using batch experiments: i) the colloidal stability of Ag NP increases through sorption of soil-borne dissolved organic matter (DOM) and thus inhibits aggregation; ii) the presence of DOM suppresses Ag oxidation; iii) the surface charge of Ag NP governs sorption onto soil particles. Citrate-stabilized and bare Ag NPs were equilibrated with (colloid-free) soil solution extracted from a floodplain soil for 24h. Nanoparticles were removed through centrifugation. Concentrations of free Ag ions and DOC, the specific UV absorbance at a wavelength of 254 nm, and the absorption ratio α254/α410 were determined in the supernatant. Nanoparticle aggregation was studied using time-resolved dynamic light scattering (DLS) measurement following the addition of soil solution and 1.5mM Ca(2+) solution. To study the effect of surface charge on the adsorption of Ag NP onto soil particles, bare and citrate-stabilized Ag NP, differing in the zeta potential, were equilibrated with silt at a solid-to-solution ratio of 1:10 and an initial Ag concentration range of 30 to 320 µg/L. Results showed that bare Ag NPs sorb organic matter, with short-chained organic matter being preferentially adsorbed over long-chained, aromatic organic matter. Stabilizing effects of organic matter only come into play at higher Ag NP concentrations. Soil solution inhibits the release of Ag(+) ions, presumably due to organic matter coatings. Sorption to silt particles was very similar for the two particle types, suggesting that the surface charge does not control Ag NP sorption. Besides, sorption was much lower than in comparable studies with sand and glass surfaces.

Disaggregation of silver nanoparticle homoaggregates in a river water matrix.

Silver nanoparticles (Ag NPs) could be found in aquatic systems in the near future. Although the interplay between aggregate formation and disaggregation is an important factor for mobility, bioavailability and toxicity of Ag NPs in surface waters, the factors controlling disaggregation of Ag NP homoaggregates are still unknown. In this study, we investigated the reversibility of homoaggregation of citrate coated Ag NPs in a Rhine River water matrix. We characterized the disaggregation of Ag NP homoaggregates by ionic strength reduction and addition of Suwannee River humic acid (SRHA) in the presence of strong and weak shear forces. In order to understand the disaggregation processes, we also studied the nature of homoaggregates and their formation dynamics under the influence of SRHA, Ca(2+) concentration and nanoparticle concentration. Even in the presence of SRHA and at low particle concentrations (10 µg L(-1)), aggregates formed rapidly in filtered Rhine water. The critical coagulation concentration (CCC) of Ca(2+) in reconstituted Rhine water was 1.5 mmol L(-1) and was shifted towards higher values in the presence of SRHA. Analysis of the attachment efficiency as a function of Ca(2+) concentration showed that SRHA induces electrosteric stabilization at low Ca(2+) concentrations and cation-bridging flocculation at high Ca(2+) concentrations. Shear forces in the form of mechanical shaking or ultrasound were necessary for breaking the aggregates. Without ultrasound, SRHA also induced disaggregation, but it required several days to reach a stable size of dense aggregates still larger than the primary particles. Citrate stabilized Ag NPs may be in the form of reaction limited aggregates in aquatic systems similar to the Rhine River. The size and the structure of these aggregates will be dynamic and be determined by the solution conditions. Seasonal variations in the chemical composition of natural waters can result in a sedimentation-release cycle of engineered nanoparticles.

Zeta potential measurement as a diagnostic tool in enzyme immobilisation.

The efficiency of binding during enzyme immobilisation does not only depend on the chemical properties of the enzyme and the matrix particle, but also on their surface potential. Zeta potential quantifies the electrostatic interactions between enzyme and matrix particles, and can therefore, be used as an indicator of the binding efficiency in the enzyme immobilisation studies. In order to establish a correlation between the zeta potential and the binding efficiency, we used CALA (Candida antarctica A-type lipase) as a model protein for immobilisation on non-porous magnetic microparticles with epoxy (M-PVA E02), carboxy (M-PVA C12) and amine (M-PVA N12) terminations. We observed maximal binding of CALA onto the M-PVA N12 beads, due to the electrostatic attraction between negatively charged protein and carrier particles with slightly positive zeta potential. The binding of CALA was lower when M-PVA E02 beads were used, followed by M-PVA C12 beads. The decreasing binding efficiency was obviously the result of increasing electrostatic repulsion between the interaction partners. This could be correlated to the increasing negative zeta potential of the magnetic particles. Moreover, the medium of suspension of the particles also makes a significant difference. We found highest specific activity of the lipase immobilised on M-PVA E02 beads in a medium concentrated buffer (0.3M). The results demonstrate a clear correlation between zeta potential and binding efficiency but no correlation between the bead related specific activity and the zeta potential. These findings are advocating the possibility of using the zeta potential as a diagnostic tool in enzyme immobilisation.