Biotic and abiotic interactions in aquatic microcosms determine fate and toxicity of Ag nanoparticles. Part 1. Aggregation and dissolution

To better understand their fate and toxicity in aquatic environments, we compared the aggregation and dissolution behavior of gum arabic (GA) and polyvinylpyrrolidone (PVP) coated Ag nanoparticles (NPs) in aquatic microcosms. There were four microcosm types: surface water; water and sediment; water and aquatic plants; or water, sediment, and aquatic plants. Dissolution and aggregation behavior of AgNPs were examined using ultracentrifugation, ultrafiltration, and asymmetrical flow field flow fractionation coupled to ultraviolet-visible spectroscopy, dynamic and static laser light scattering, and inductively coupled plasma mass spectrometry. Plants released dissolved organic matter (DOM) into the water column either through active or passive processes in response to Ag exposure. This organic matter fraction readily bound Ag ions. The plant-derived DOM had the effect of stabilizing PVP-AgNPs as primary particles, but caused GA-AgNPs to be removed from the water column, likely by dissolution and binding of released Ag ions on sediment and plant surfaces. The destabilization of the GA-AgNPs also corresponded with X-ray absorption near edge spectroscopy results which suggest that 22-28% of the particulate Ag was associated with thiols and 5-14% was present as oxides. The results highlight the potential complexities of nanomaterial behavior in response to biotic and abiotic modifications in ecosystems, and may help to explain differences in toxicity of Ag observed in realistic exposure media compared to simplified laboratory exposures.     

Differential effect of common ligands and molecular oxygen on antimicrobial activity of silver nanoparticles versus silver ions

The antibacterial activity of silver nanoparticles (AgNPs) is partially due to the release of Ag(+), although discerning the contribution of AgNPs vs Ag(+) is challenging due to their common co-occurrence. We discerned the toxicity of Ag(+) versus a commercially available AgNP (35.4 ± 5.1 nm, coated with amorphous carbon) by conducting antibacterial assays under anaerobic conditions that preclude Ag? oxidation, which is a prerequisite for Ag(+) release. These AgNPs were 20× less toxic to E. coli than Ag(+) (EC??: 2.04 ± 0.07 vs 0.10 ± 0.01 mg/L), and their toxicity increased 2.3-fold after exposure to air for 0.5 h (EC??: 0.87 ± 0.03 mg/L) which promoted Ag(+) release. No significant difference in Ag(+) toxicity was observed between anaerobic and aerobic conditions, which rules out oxidative stress by ROS as an important antibacterial mechanism for Ag(+). The toxicity of Ag(+) (2.94 μmol/L) was eliminated by equivalent cysteine or sulfide; the latter exceeded the solubility product equilibrium constant (K(sp)), which is conducive to silver precipitation. Equivalent chloride and phosphate concentrations also reduced Ag(+) toxicity without exceeding K(sp). Thus, some common ligands can hinder the bioavailability and mitigate the toxicity of Ag(+) at relatively low concentrations that do not induce silver precipitation. Furthermore, low concentrations of chloride (0.1 mg/L) mitigated the toxicity of Ag(+) but not that of AgNPs, suggesting that previous reports of higher AgNPs toxicity than their equivalent Ag(+) concentration might be due to the presence of common ligands that preferentially decrease the bioavailability and toxicity of Ag(+). Overall, these results show that the presence of O? or common ligands can differentially affect the toxicity of AgNPs vs Ag(+), and underscore the importance of water chemistry in the mode of action of AgNPs.     

Aggregation and dissolution of silver nanoparticles in natural surface water

This study investigated aggregation and silver release of silver nanoparticles suspended in natural water in the absence and presence of artificial sun light. The influence of the capping layer was investigated using uncoated particles and particles coated with citrate or Tween 80. The experiments were conducted over 15 days in batch mode using a river water matrix. Silver release was monitored over this time while the aggregation state and morphological changes of the silver nanoparticles were tracked using dynamic light scattering and transmission electron microscopy. Results indicate sterically dispersed particles coated with Tween released silver quicker than did bare- and citrate-coated particles, which rapidly aggregated. A dissolved silver concentration of 40 μg/L was reached after just 6 h in the Tween-coated particle systems, accounting for ca. 3% of the total silver. Similar levels of dissolved silver were reached in the uncoated and citrate-coated systems at the end of the 15 days. Silver release was not significantly impacted by the artificial sun light; however, the light (and citrate) imparted significant morphological changes to the particles. Their impact was masked by aggregation, which seemed to be the controlling process in this study.     

Bioaccumulation dynamics and modeling in an estuarine invertebrate following aqueous exposure to nanosized and dissolved silver

Predicting the environmental impact of engineered nanomaterials (ENMs) is increasingly important owing to the prevalence of emerging nanotechnologies. We derived waterborne uptake and efflux rate constants for the estuarine snail, Peringia ulvae, exposed to dissolved Ag (AgNO(3)) and silver nanoparticles (Ag NPs), using biodynamic modeling. Uptake rates demonstrated that dissolved Ag is twice as bioavailable as Ag in nanoparticle form. Biphasic loss dynamics revealed the faster elimination of Ag from Ag NPs at the start of depuration, but similar slow efflux rate constants. The integration of biodynamic parameters into our model accurately predicted Ag tissue burdens during chronic exposure with 85% of predicted values within a factor of 2 of observed values. Zeta potentials for the Ag NPs were lower in estuarine waters than in waters of less salinity; and uptake rates in P. ulvae were slower than reported for the freshwater snail Lymnaea stagnalis in similar experiments. This suggests aggregation of Ag NPs occurs in estuarine waters and reduces, but does not eliminate, bioavailability of Ag from the Ag NPs. Biodynamic modeling provides an effective methodology to determine bioavailable metal concentrations (originating from metal and metal-oxide nanoparticles) in the environment and may aid future ENM risk assessment.     

Intracellular silver accumulation in Chlamydomonas reinhardtii upon exposure to carbonate coated silver nanoparticles and silver nitrate

The intracellular silver accumulation ({Ag}(in)) upon exposure to carbonate coated silver nanoparticles (AgNP, 0.5-10 μM, average diameter 29 nm) and silver nitrate (20-500 nM) was examined in the wild type and in the cell wall free mutant of the green alga Chlamydomonas reinhardtii at pH 7.5. The {Ag}(in) was measured over time up to 1 h after a wash procedure to remove silver ions (Ag(+)) and AgNP from the algal cell surface. The {Ag}(in) increased with increasing exposure time and with increasing AgNP and AgNO(3) concentrations in the exposure media, reaching steady-state concentrations between 10(-5) and 10(-3) mol L(cell)(-1). According to estimated kinetic parameters, high Ag(+) bioconcentration factors were calculated (>10(3) L L(cell)(-1)). Higher accumulation rate constants were assessed in the cell wall free mutant, indicating a protective role of the cell wall in limiting Ag(+) uptake. The bioavailability of AgNP was calculated to be low in both strains relative to Ag(+), suggesting that AgNP internalization across the cell membrane was limited.     

Mechanism of silver nanoparticle toxicity is dependent on dissolved silver and surface coating in Caenorhabditis elegans

The rapidly increasing use of silver nanoparticles (Ag NPs) in consumer products and medical applications has raised ecological and human health concerns. A key question for addressing these concerns is whether Ag NP toxicity is mechanistically unique to nanoparticulate silver, or if it is a result of the release of silver ions. Furthermore, since Ag NPs are produced in a large variety of monomer sizes and coatings, and since their physicochemical behavior depends on the media composition, it is important to understand how these variables modulate toxicity. We found that a lower ionic strength medium resulted in greater toxicity (measured as growth inhibition) of all tested Ag NPs to Caenorhabditis elegans and that both dissolved silver and coating influenced Ag NP toxicity. We found a linear correlation between Ag NP toxicity and dissolved silver, but no correlation between size and toxicity. We used three independent and complementary approaches to investigate the mechanisms of toxicity of differentially coated and sized Ag NPs: pharmacological (rescue with trolox and N-acetylcysteine), genetic (analysis of metal-sensitive and oxidative stress-sensitive mutants), and physicochemical (including analysis of dissolution of Ag NPs). Oxidative dissolution was limited in our experimental conditions (maximally 15% in 24 h) yet was key to the toxicity of most Ag NPs, highlighting a critical role for dissolved silver complexed with thiols in the toxicity of all tested Ag NPs. Some Ag NPs (typically less soluble due to size or coating) also acted via oxidative stress, an effect specific to nanoparticulate silver. However, in no case studied here was the toxicity of a Ag NP greater than would be predicted by complete dissolution of the same mass of silver as silver ions.     

Precipitation and growth of zinc sulfide nanoparticles in the presence of thiol-containing natural organic ligands

In sulfidic aquatic systems, metal sulfides can control the mobility and bioavailability of trace metal pollutants such as zinc, mercury, and silver. Nanoparticles of ZnS and other metal sulfides are known to exist in oxic and anoxic waters. However, the processes that lead to their persistence in the aquatic environment are relatively unknown. The objective of this study was to evaluate the importance of dissolved natural organics in stabilizing nanoparticulate ZnS that precipitates under environmentally relevant conditions. Precipitation and growth of ZnS particles were investigated in the presence of dissolved humic acid and low-molecular weight organic acids that are prevalent in sediment porewater. Dynamic light scattering was used to monitor the hydrodynamic diameter of particles precipitating in laboratory solutions. Zn speciation was also measured by filtering the ZnS solutions (< 0.2 microm) and using anodic stripping voltammetry to confirm that Zn was coordinated to sulfide during the precipitation experiments and not to the dissolved organic ligands. X-ray photoelectron spectroscopy and electron microscopy were used to confirm that amorphous particles containing Zn and S were precipitating in the suspensions. Observed growth rates of ZnS particles varied by orders of magnitude, depending on the type and concentration of organic ligand in solution. In the presence of humic acid and thiol-containing ligands (cysteine, glutathione, and thioglycolate), observed growth rates decreased by 1-3 orders of magnitude relative to controls without the ligands. In contrast, growth rates of the particles were consistently within 1 order of magnitude of the ligand-free control when oxygen- and amine-containing ligands (oxalate, serine, and glycolate) were present Furthermore, particle growth rates decreased with an increase in thiol concentration and increased with NaNO3 electrolyte concentration. These studies suggest that specific surface interactions with thiol-containing organics may be one factor that contributes to the persistence of naturally occurring and anthropogenic nanoparticles of ZnS and other metal sulfides in the aquatic environment.     

More than the ions: the effects of silver nanoparticles on Lolium multiflorum

Silver nanoparticles (AgNPs) are increasingly used as antimicrobial additives in consumer products and may have adverse impacts on organisms when they inadvertently enter ecosystems. This study investigated the uptake and toxicity of AgNPs to the common grass, Lolium multiflorum. We found that root and shoot Ag content increased with increasing AgNP exposures. AgNPs inhibited seedling growth. While exposed to 40 mg L(-1) GA-coated AgNPs, seedlings failed to develop root hairs, had highly vacuolated and collapsed cortical cells and broken epidermis and rootcap. In contrast, seedlings exposed to identical concentrations of AgNO(3) or supernatants of ultracentrifuged AgNP solutions showed no such abnormalities. AgNP toxicity was influenced by total NP surface area with smaller AgNPs (6 nm) more strongly affecting growth than did similar concentrations of larger (25 nm) NPs for a given mass. Cysteine (which binds Ag(+)) mitigated the effects of AgNO(3) but did not reduce the toxicity of AgNP treatments. X-ray spectro-microscopy documented silver speciation within exposed roots and suggested that silver is oxidized within plant tissues. Collectively, this study suggests that growth inhibition and cell damage can be directly attributed either to the nanoparticles themselves or to the ability of AgNPs to deliver dissolved Ag to critical biotic receptors.     

Sulfidation of silver nanoparticles decreases Escherichia coli growth inhibition

Sulfidation of metallic nanoparticles such as silver nanoparticles (AgNPs) released to the environment may be an important detoxification mechanism. Two types of AgNPs-an engineered polydisperse and aggregated AgNP powder, and a laboratory-synthesized, relatively monodisperse AgNP aqueous dispersion-were studied. The particles were sulfidized to varying degrees and characterized to determine the effect of initial AgNP polydispersity and aggregation state on AgNP sulfidation, and then exposed to Escherichia coli to determine if the degree of sulfidation of pristine AgNPs affects growth inhibition of bacteria. The extent of sulfidation was found to depend on the HS(-)/Ag ratio. However, for the same reaction times, the more monodisperse particles were fully transformed to Ag(2)S, and the polydisperse, aggregated particles were not fully sulfidized, thus preserving the toxic potential of Ag(0) in the aggregates. A higher Ag(2)S:Ag(0) ratio in the sulfidized nanoparticles resulted in less growth inhibition of E. coli over 6 h of exposure. These results suggest that the initial properties of AgNPs can affect sulfidation products, which in turn affect microbial growth inhibition, and that these properties should be considered in assessing the environmental impact of AgNPs.     

Fractionating nanosilver: importance for determining toxicity to aquatic test organisms

This investigation applied novel techniques for characterizing and fractionating nanosilver particles and aggregates and relating these measurements to toxicological endpoints. The acute toxicity of eight nanosilver suspensions of varying primary particle sizes (10-80 nm) and coatings (citrate, polyvinylpyrrolidone, EDTA, proprietary) was assessed using three aquatic test organisms (Daphnia magna, Pimephales promelas, Pseudokirchneriella subcapitata). When 48-h lethal median concentrations (LC50) were expressed as total silver, both D. magna and P. promelas were significantly more sensitive to ionic silver (Ag(+)) as AgNO(3) (mean LC50 = 1.2 and 6.3 μg/L, respectively) relative to a wide range in LC50 values determined for the nanosilver suspensions (2 -126 μg/L). However, when LC50 values for nanosilver suspensions were expressed as fractionated nanosilver (Ag(+) and/or <4 nm particles), determined by ultracentrifugation of particles and confirmed field-flow-fractograms, the LC50 values (0.3-5.6 μg/L) were comparable to the values obtained for ionic Ag(+) as AgNO(3). These results suggest that dissolved Ag(+) plays a critical role in acute toxicity and underscores the importance of characterizing dissolved fractions in nanometal suspensions.     

Environmental transformations of silver nanoparticles: impact on stability and toxicity

Silver nanoparticles (Ag-NPs) readily transform in the environment, which modifies their properties and alters their transport, fate, and toxicity. It is essential to consider such transformations when assessing the potential environmental impact of Ag-NPs. This review discusses the major transformation processes of Ag-NPs in various aqueous environments, particularly transformations of the metallic Ag cores caused by reactions with (in)organic ligands, and the effects of such transformations on physical and chemical stability and toxicity. Thermodynamic arguments are used to predict what forms of oxidized silver will predominate in various environmental scenarios. Silver binds strongly to sulfur (both organic and inorganic) in natural systems (fresh and sea waters) as well as in wastewater treatment plants, where most Ag-NPs are expected to be concentrated and then released. Sulfidation of Ag-NPs results in a significant decrease in their toxicity due to the lower solubility of silver sulfide, potentially limiting their short-term environmental impact. This review also discusses some of the major unanswered questions about Ag-NPs, which, when answered, will improve predictions about their potential environmental impacts. Research needed to address these questions includes fundamental molecular-level studies of Ag-NPs and their transformation products, particularly Ag(2)S-NPs, in simplified model systems containing common (in)organic ligands, as well as under more realistic environmental conditions using microcosm/mesocosm-type experiments. Toxicology studies of Ag-NP transformation products, including different states of aggregation and sulfidation, are also required. In addition, there is the need to characterize the surface structures, compositions, and morphologies of Ag-NPs and Ag(2)S-NPs to the extent possible because they control properties such as solubility and reactivity.     

Sulfidation processes of PVP-coated silver nanoparticles in aqueous solution: impact on dissolution rate

Despite the increasing use of silver nanoparticles (Ag-NPs) in nanotechnology and their toxicity to invertebrates, the transformations and fate of Ag-NPs in the environment are poorly understood. This work focuses on the sulfidation processes of PVP-coated Ag-NPs, one of the most likely corrosion phenomena that may happen in the environment. The sulfur to Ag-NPs ratio was varied in order to control the extent of Ag-NPs transformation to silver sulfide (Ag?S). A combination of synchrotron-based X-ray Diffraction (XRD) and Extended X-ray Absorption Fine Structure spectroscopy shows the increasing formation of Ag?S with an increasing sulfur to Ag-NPs ratio. TEM observations show that Ag?S forms nanobridges between the Ag-NPs leading to chain-like structures. In addition, sulfidation strongly affects surface properties of the Ag-NPs in terms of surface charge and dissolution rate. Both may affect the reactivity, transport, and toxicity of Ag-NPs in soils. In particular, the decrease of dissolution rate as a function of sulfide exposure may strongly limit Ag-NPs toxicity since released Ag? ions are known to be a major factor in the toxicity of Ag-NPs.     

Cellular internalization of silver nanoparticles in gut epithelia of the estuarine polychaete Nereis diversicolor

Silver nanoparticles (AgNPs) are widely used which may result in environmental impacts, notably within aquatic ecosystems. As estuarine sediments are sinks for numerous pollutants, but also habitat and food for deposit feeders such as Nereis diversicolor, ingested sediments must be investigated as an important route of uptake for NPs. N. diversicolor were fed sediment spiked with either citrate capped AgNPs (30 ± 5 nm) or aqueous Ag for 10 days. Postexposure AgNPs were observed in the lumen of exposed animals, and three lines of evidence indicated direct internalization of AgNPs into the gut epithelium. With TEM, electron-dense particles resembling AgNPs were observed associated with the apical plasma membrane, in endocytotic pits and in endosomes. Energy dispersive X-ray analysis (EDX) confirmed the presence of Ag in these particles, which were absent in controls. Subcellular fractionation revealed that Ag accumulated from AgNPs was predominantly associated with inorganic granules, organelles, and the heat denatured proteins; whereas dissolved Ag was localized to the metallothionein fraction. Collectively, these results indicate separate routes of cellular internalization and differing in vivo fates of Ag delivered in dissolved and NP form. For AgNPs an endocytotic pathway appears to be a key route of cellular uptake.     

Exposure to copper nanoparticles causes gill injury and acute lethality in zebrafish (Danio rerio)

Increasing use of metallic nanomaterials is likely to result in release of these particles into aqueous environments; however, it is unclear if these materials present a hazard to aquatic organisms. Because some dissolution of metal particles will occur, it is important to distinguish effects of nanoparticulates from dissolved metals. To address this issue, acute toxicity of soluble copper and 80 nm copper nanoparticle suspensions were examined in zebrafish. The results demonstrate that nanocopper is acutely toxic to zebrafish, with a 48 h LC50 concentration of 1.5 mg/L. Rapid aggregation of copper nanoparticles occurred after suspension in water, resulting in 50-60% of added mass leaving the water column. While dissolution of particulate copper occurred, it was insufficient to explain the mortality in nanocopper exposures. Histological and biochemical analysis revealed that the gill was the primary target organ for nanocopper. To further investigate the effects of nanocopper on the gill, zebrafish were exposed to 100 microg/L of nanocopper or to the concentration of soluble copper matching that present due to dissolution of the particles. Under these conditions, nanocopper produced different morphological effects and global gene expression patterns in the gill than soluble copper, clearly demonstrating that the effects of nanocopper on gill are not mediated solely by dissolution.     

Comparative toxicity of nanoparticulate ZnO, bulk ZnO, and ZnCl2 to a freshwater microalga (Pseudokirchneriella subcapitata): the importance of particle solubility

Metal oxide nanoparticles are finding increasing application in various commercial products, leading to concerns for their environmental fate and potential toxicity. It is generally assumed that nanoparticles will persist as small particles in aquatic systems and that their bioavailability could be significantly greater than that of larger particles. The current study using nanoparticulate ZnO (ca. 30 nm) has shown that this is not always so. Particle characterization using transmission electron microscopy and dynamic light scattering techniques showed that particle aggregation is significant in a freshwater system, resulting in flocs ranging from several hundred nanometers to several microns. Chemical investigations using equilibrium dialysis demonstrated rapid dissolution of ZnO nanoparticles in a freshwater medium (pH 7.6), with a saturation solubility in the milligram per liter range, similar to that of bulk ZnO. Toxicity experiments using the freshwater alga Pseudokirchneriella subcapitata revealed comparable toxicity for nanoparticulate ZnO, bulk ZnO, and ZnCl2, with a 72-h IC50 value near 60 microg Zn/ L, attributable solely to dissolved zinc. Care therefore needs to be taken in toxicity testing in ascribing toxicity to nanoparticles per se when the effects may be related, at least in part, to simple solubility.     

Effects of humic substances on precipitation and aggregation of zinc sulfide nanoparticles

Nanoparticulate metal sulfides such as ZnS can influence the transport and bioavailability of pollutant metals in anaerobic environments. The aim of this work was to investigate how the composition of dissolved natural organic matter (NOM) influences the stability of zinc sulfide nanoparticles as they nucleate and aggregate in water with dissolved NOM. We compared NOM fractions that were isolated from several surface waters and represented a range of characteristics including molecular weight, type of carbon, and ligand density. Dynamic light scattering was employed to monitor the growth and aggregation of Zn-S-NOM nanoparticles in supersaturated solutions containing dissolved aquatic humic substances. The NOM was observed to reduce particle growth rates, depending on solution variables such as type and concentration of NOM, monovalent electrolyte concentration, and pH. The rates of growth increased with increasing ionic strength, indicating that observed growth rates primarily represented aggregation of charged Zn-S-NOM particles. Furthermore, the observed rates decreased with increasing molecular weight and aromatic content of the NOM fractions, while carboxylate and reduced sulfur content had little effect. Differences between NOM were likely due to properties that increased electrosteric hindrances for aggregation. Overall, results of this study suggest that the composition and source of NOM are key factors that contribute to the stabilization and persistence of zinc sulfide nanoparticles in the aquatic environment.     

Stability, bioavailability, and bacterial toxicity of ZnO and iron-doped ZnO nanoparticles in aquatic media

The stability and bioavailability of nanoparticles is governed by the interfacial properties that nanoparticles acquire when immersed in a particular aquatic media as well as the type of organism or cell under consideration. Herein, high-throughput screening (HTS) was used to elucidate ZnO nanoparticle stability, bioavailability, and antibacterial mechanisms as a function of iron doping level (in the ZnO nanoparticles), aquatic chemistry, and bacterial cell type. ζ-Potential and aggregation state of dispersed ZnO nanoparticles was strongly influenced by iron doping in addition to electrolyte composition and dissolved organic matter; however, bacterial inactivation by ZnO nanoparticles was most significantly influenced by Zn(2+) ions dissolution, cell type, and organic matter. Nanoparticle IC(50) values determined for Bacillus subtilis and Escherichia coli were on the order of 0.3-0.5 and 15-43 mg/L (as Zn(2+)), while the IC(50) for Zn(2+) tolerant Pseudomonas putida was always >500 mg/L. Tannic acid decreased toxicity of ZnO nanoparticles more than humic, fulvic, and alginic acid, because it complexed the most free Zn(2+) ions, thereby reducing their bioavailability. These results underscore the complexities and challenges regulators face in assessing potential environmental impacts of nanotechnology; however, the high-throughput and combinatorial methods employed promise to rapidly expand the knowledge base needed to develop an appropriate risk assessment framework.     

Behavior of metallic silver nanoparticles in a pilot wastewater treatment plant

We investigated the behavior of metallic silver nanoparticles (Ag-NP) in a pilot wastewater treatment plant (WWTP) fed with municipal wastewater. The treatment plant consisted of a nonaerated and an aerated tank and a secondary clarifier. The average hydraulic retention time including the secondary clarifier was 1 day and the sludge age was 14 days. Ag-NP were spiked into the nonaerated tank and samples were collected from the aerated tank and from the effluent. Ag concentrations determined by inductively coupled plasma-mass spectrometry (ICP-MS) were in good agreement with predictions based on mass balance considerations. Transmission electron microscopy (TEM) analyses confirmed that nanoscale Ag particles were sorbed to wastewater biosolids, both in the sludge and in the effluent. Freely dispersed nanoscale Ag particles were only observed in the effluent during the initial pulse spike. X-ray absorption spectroscopy (XAS) measurements indicated that most Ag in the sludge and in the effluent was present as Ag(2)S. Results from batch experiments suggested that Ag-NP transformation to Ag(2)S occured in the nonaerated tank within less than 2 h. Physical and chemical transformations of Ag-NP in WWTPs control the fate, the transport and also the toxicity and the bioavailability of Ag-NP and therefore must be considered in future risk assessments.     

Methylation of mercury by bacteria exposed to dissolved, nanoparticulate, and microparticulate mercuric sulfides

The production of the neurotoxic methylmercury in the environment is partly controlled by the bioavailability of inorganic divalent mercury (Hg(II)) to anaerobic bacteria that methylate Hg(II). In sediment porewater, Hg(II) associates with sulfides and natural organic matter to form chemical species that include organic-coated mercury sulfide nanoparticles as reaction intermediates of heterogeneous mineral precipitation. Here, we exposed two strains of sulfate-reducing bacteria to three forms of inorganic mercury: dissolved Hg and sulfide, nanoparticulate HgS, and microparticulate HgS. The bacteria cultures exposed to HgS nanoparticles methylated mercury at a rate slower than cultures exposed to dissolved forms of mercury. However, net methylmercury production in cultures exposed to nanoparticles was 6 times greater than in cultures treated with microscale particles, even when normalized to specific surface area. Furthermore, the methylation potential of HgS nanoparticles decreased with storage time of the nanoparticles in their original stock solution. In bacteria cultures amended with nano-HgS from a 16 h-old nanoparticle stock, 6-10% of total mercury was converted to methylmercury after one day. In contrast, 2-4% was methylated in cultures amended with nano-HgS that was aged for 3 days or 1 week. The methylation of mercury derived from nanoparticles (in contrast to the larger particles) would not be predicted by equilibrium speciation of mercury in the aqueous phase (<0.2 μm) and was possibly caused by the disordered structure of nanoparticles that facilitated release of chemically labile mercury species immediately adjacent to cell surfaces. Our results add new dimensions to the mechanistic understanding of mercury methylation potential by demonstrating that bioavailability is related to the geochemical intermediates of rate-limited mercury sulfide precipitation reactions. These findings could help explain observations that the "aging" of mercury in sediments reduces its methylation potential and provide a basis for assessing and remediating methylmercury hotspots in the environment.     

Silver nanoparticle-algae interactions: oxidative dissolution, reactive oxygen species generation and synergistic toxic effects

The short-term toxicity of citrate-stabilized silver nanoparticles (AgNPs) and ionic silver Ag(I) to the ichthyotoxic marine raphidophyte Chattonella marina has been examined using the fluorometric indicator alamarBlue. Aggregation and dissolution of AgNPs occurred after addition to GSe medium while uptake of dissolved Ag(I) occurred in the presence of C. marina. Based on total silver mass, toxicity was much higher for Ag(I) than for AgNPs. Cysteine, a strong Ag(I) ligand, completely removed the inhibitory effects of Ag(I) and AgNPs on the metabolic activity of C. marina, suggesting that the toxicity of AgNPs was due to the release of Ag(I). Synergistic toxic effects of AgNPs/Ag(I) and C. marina to fish gill cells were observed with these effects possibly attributable to enhancement in the generation of reactive oxygen species by C. marina on exposure of the organism to silver.