Chronic Engineered Nanoparticle Additions Alter Insect Emergence and Result in Metal Flux from Aquatic Ecosystems into Riparian Food Webs.

Freshwater ecosystems are exposed to engineered nanoparticles (NPs) through discharge from wastewater and agricultural runoff. We conducted a 9-month mesocosm experiment to examine the combined effects of chronic NP additions on insect emergence and insect-mediated contaminant flux to riparian spiders. Two NPs (copper, gold, plus controls) were crossed by two levels of nutrients in 18 outdoor mesocosms open to natural insect and spider colonization. We collected adult insects and two riparian spider genera, Tetragnatha and Dolomedes, for 1 week on a monthly basis. We estimated a significant decrease in cumulative insect emergence of 19% and 24% after exposure to copper and gold NPs, irrespective of nutrient level. NP treatments led to elevated copper and gold tissue concentrations in adult insects, which resulted in terrestrial fluxes of metals. These metal fluxes were associated with increased gold and copper tissue concentrations for both spider genera. We also observed about 25% fewer spiders in the NP mesocosms, likely due to reduced insect emergence and/or NP toxicity. These results demonstrate the transfer of NPs from aquatic to terrestrial ecosystems via emergence of aquatic insects and predation by riparian spiders, as well as significant reductions in insect and spider abundance in response to NP additions.

Copper and Gold Nanoparticles Increase Nutrient Excretion Rates of Primary Consumers.

Freshwater ecosystems are exposed to engineered nanoparticles through municipal and industrial wastewater-effluent discharges and agricultural nonpoint source runoff. Because previous work has shown that engineered nanoparticles from these sources can accumulate in freshwater algal assemblages, we hypothesized that nanoparticles may affect the biology of primary consumers by altering the processing of two critical nutrients associated with growth and survivorship, nitrogen and phosphorus. We tested this hypothesis by measuring the excretion rates of nitrogen and phosphorus of Physella acuta, a ubiquitous pulmonate snail that grazes heavily on periphyton, exposed to either copper or gold engineered nanoparticles for 6 months in an outdoor wetland mesocosm experiment. Chronic nanoparticle exposure doubled nutrient excretion when compared to the control. Gold nanoparticles increased nitrogen and phosphorus excretion rates more than copper nanoparticles, but overall, both nanoparticles led to higher consumer excretion, despite contrasting particle stability and physiochemical properties. Snails in mesocosms enriched with nitrogen and phosphorus had overall higher excretion rates than ones in ambient (no nutrients added) mesocosms. Stimulation patterns were different between nitrogen and phosphorus excretion, which could have implications for the resulting nutrient ratio in the water column. These results suggest that low concentrations of engineered nanoparticles could alter the metabolism of consumers and increase consumer-mediated nutrient recycling rates, potentially intensifying eutrophication in aquatic systems, for example, the increased persistence of algal blooms as observed in our mesocosm experiment.

Differential Reactivity of Copper- and Gold-Based Nanomaterials Controls Their Seasonal Biogeochemical Cycling and Fate in a Freshwater Wetland Mesocosm.

Reliable predictions of the environmental fate and risk of engineered nanomaterials (ENMs) require a better understanding of ENM reactivity in complex, biologically active systems for chronic low-concentration exposure scenarios. Here, simulated freshwater wetland mesocosms were dosed with ENMs to assess how their reactivity and seasonal changes in environmental parameters influence ENM fate in aquatic systems. Copper-based ENMs (Kocide), known to dissolve in water, and gold nanoparticles (AuNPs), stable against dissolution in the absence of specific ligands, were added weekly to mesocosm waters for 9 months. Metal accumulation and speciation changes in the different environmental compartments were assessed over time. Copper from Kocide rapidly dissolved likely associating with organic matter in the water column, transported to terrestrial soils and deeper sediment where it became associated with organic or sulfide phases. In contrast, Au accumulated on/in the macrophytes where it oxidized and transferred over time to surficial sediment. A dynamic seasonal accumulation and metal redox cycling were found between the macrophyte and the surficial sediment for AuNPs. These results demonstrate the need for experimental quantification of how the biological and chemical complexity of the environment, combined with their seasonal variations, drive the fate of metastable ENMs.

Conserved Microbial Toxicity Responses for Acute and Chronic Silver Nanoparticle Treatments in Wetland Mesocosms.

Most studies of bacterial exposure to environmental contaminants focus on acute treatments; however, the impacts of single, high-dose exposures on microbial communities may not readily be extended to the more likely scenario of chronic, low-dose contaminant exposures. Here, in a year-long, wetland mesocosm experiment, we compared microbial community responses to pulse (single 450 mg dose of silver) and chronic (weekly 8.7 mg doses of silver for 1 year) silver nanoparticle (Ag0 NP) treatments, as well as a chronic treatment of "aged" sulfidized silver nanoparticles (Ag2S NPs). While mesocosms exposed to Ag2S NPs never differed significantly from the controls, both Ag0 NP treatments exhibited reduced microbial diversity and altered community composition; however, the effects differed in timing, duration, and magnitude. Microbial community-level impacts in the acute Ag0 NP treatment were apparent only within the first weeks and then converged on the control mesocosm composition, while chronic exposure effects were observed several months after exposures began, likely due to interactive effects of nanoparticle toxicity and winter environmental conditions. Notably, there was a high level of overlap in the taxa which exhibited significant declines (>10×) in both treatments, suggesting a conserved toxicity response for both pulse and chronic exposures. Thus, this research suggests that complex, but short-term, acute toxicological studies may provide critical, cost-effective insights into identifying microbial taxa sensitive to long-term chronic exposures to Ag NPs.

Engineered nanoparticles interact with nutrients to intensify eutrophication in a wetland ecosystem experiment.

Despite the rapid rise in diversity and quantities of engineered nanomaterials produced, the impacts of these emerging contaminants on the structure and function of ecosystems have received little attention from ecologists. Moreover, little is known about how manufactured nanomaterials may interact with nutrient pollution in altering ecosystem productivity, despite the recognition that eutrophication is the primary water quality issue in freshwater ecosystems worldwide. In this study, we asked two main questions: (1) To what extent do manufactured nanoparticles affect the biomass and productivity of primary producers in wetland ecosystems? (2) How are these impacts mediated by nutrient pollution? To address these questions, we examined the impacts of a citrate-coated gold nanoparticle (AuNPs) and of a commercial pesticide containing Cu(OH)2 nanoparticles (CuNPs) on aquatic primary producers under both ambient and enriched nutrient conditions. Wetland mesocosms were exposed repeatedly with low concentrations of nanoparticles and nutrients over the course of a 9-month experiment in an effort to replicate realistic field exposure scenarios. In the absence of nutrient enrichment, there were no persistent effects of AuNPs or CuNPs on primary producers or ecosystem productivity. However, when combined with nutrient enrichment, both NPs intensified eutrophication. When either of these NPs were added in combination with nutrients, algal blooms persisted for >50 d longer than in the nutrient-only treatment. In the AuNP treatment, this shift from clear waters to turbid waters led to large declines in both macrophyte growth and rates of ecosystem gross primary productivity (average reduction of 52% ± 6% and 92% ± 5%, respectively) during the summer. Our results suggest that nutrient status greatly influences the ecosystem-scale impact of two emerging contaminants and that synthetic chemicals may be playing an under-appreciated role in the global trends of increasing eutrophication. We provide evidence here that chronic exposure to Au and Cu(OH)2 nanoparticles at low concentrations can intensify eutrophication of wetlands and promote the occurrence of algal blooms.

Stress Responses of Aquatic Plants to Silver Nanoparticles.

Silver nanoparticles (AgNPs) are increasingly used in consumer products, biotechnology, and medicine, and are released into aquatic ecosystems through wastewater discharge. This study investigated the phytotoxicity of AgNPs to aquatic plants, Egeria densa and Juncus effusus by measuring physiologic and enzymatic responses to AgNP exposure under three release scenarios: two chronic (8.7 mg, weekly) exposures to either zerovalent AgNPs or sulfidized silver nanoparticles; and a pulsed (450 mg, one-time) exposure to zerovalent AgNPs. Plant enzymatic and biochemical stress responses were assessed using superoxide dismutase (SOD) and peroxidase (POD) activity, malondialdehyde (MDA) concentrations and chlorophyll content as markers of defense and phytotoxicity, respectively. The high initial pulse treatment resulted in rapid changes in physiological characteristics and silver concentration in plant tissue at the beginning of each AgNPs exposure (6 h, 36 h, and 9 days), while continuous AgNP and sulfidized AgNP chronic treatments gave delayed responses. Both E. densa and J. effusus enhanced their tolerance to AgNPs toxicity by increasing POD and SOD activities to scavenge free radicals but at different growth phases. Chlorophyll did not change. After AgNPs exposure, MDA, an index of membrane damage, was higher in submerged E. densa than emergent J. effusus, which suggested that engineered nanoparticles exerted more stress to submerged macrophytes.

Dosing, Not the Dose: Comparing Chronic and Pulsed Silver Nanoparticle Exposures.

The environmental impacts of manufactured nanoparticles are often studied using high-concentration pulse-additions of freshly synthesized nanoparticles, while predicted releases are characterized by chronic low-concentration additions of weathered particles. To test the effects in wetlands of addition rate and nanoparticle speciation on water column silver concentrations, ecosystem impacts, and silver accumulation by biota, we conducted a year-long mesocosm experiment. We compared a pulse addition of Ag0-NPs to chronic weekly additions of either Ag0-NPs or sulfidized silver nanoparticles. The initially high water column silver concentrations in the pulse treatment declined such that after 4 weeks it was lower on average than in the two chronic treatments. While the pulse caused a marked increase in dissolved methane in the first week of the experiment, the chronic treatments had smaller increases in methane concentration that were more prolonged between weeks 28-45. Much like water column silver, most organisms in chronic treatments had comparable silver concentrations to the pulse treatment after only 4 weeks, and all but one organism had similar or higher concentrations than the pulse treatment after one year. Pulse exposures thus both overestimate the intensity of short-term exposures and effects and underestimate the more realistic long-term exposure, ecosystem effects, and accumulation seen in chronic exposures.

Uptake and Distribution of Silver in the Aquatic Plant Landoltia punctata (Duckweed) Exposed to Silver and Silver Sulfide Nanoparticles.

Aquatic ecosystems are expected to receive Ag0 and Ag2S nanoparticles (NPs) through anthropogenic waste streams. The speciation of silver in Ag-NPs affects their fate in ecosystems, but its influence on interactions with aquatic plants is still unclear. Here, the Ag speciation and distribution was measured in an aquatic plant, duckweed (Landoltia punctata), exposed to Ag0 or Ag2S NPs, or to AgNO3. The silver distribution in duckweed roots was visualized using synchrotron-based micro X-ray fluorescence (XRF) mapping and Ag speciation was determined using extended X-ray absorption fine structure (EXAFS) spectroscopy. Duckweed exposed to Ag2S-NPs or Ag0-NPs accumulated similar Ag concentrations despite an order of magnitude smaller dissolved Ag fraction measured in the exposure medium for Ag2S-NPs compared to Ag0-NPs. By 24 h after exposure, all three forms of silver had accumulated on and partially in the roots regardless of the form of Ag exposed to the plants. Once associated with duckweed tissue, Ag0-NPs had transformed primarily into silver sulfide and silver thiol species. This suggests that plant defenses were active within or at the root surface. The Ag2S-NPs remained as Ag2S, while AgNO3 exposure led to Ag0 and sulfur-associated Ag species in plant tissue. Thus, regardless of initial speciation, Ag was readily available to duckweed.

Importance of a nanoscience approach in the understanding of major aqueous contamination scenarios: case study from a recent coal ash spill.

A coal ash spill that occurred from an ash impoundment pond into the Dan River, North Carolina, provided a unique opportunity to study the significance and role of naturally occurring and incidental nanomaterials associated with contaminant distribution from a large-scale, acute aquatic contamination event. Besides traditional measurements of bulk watercolumn and sediment metal concentrations, the nanoparticle (NP) analyses are based on cross-flow ultrafiltration (CFUF) and advanced transmission electron microscopy (TEM) techniques. A drain pipe fed by coal ash impoundment seepage showed a high level of arsenic, with concentrations many times over the EPA limit. The majority of the arsenic was found sorbed to large aggregates dominated by incidental iron oxyhydroxide (ferrihydrite) NPs, while the remainder of the arsenic was truly dissolved. These ferrihydrites were probably formed in situ where Fe(II) was leached through subsurface flowpaths into an aerobic environment, and further act as a significant contributor to the elevated As concentrations in downstream sediments after the spill. In addition, we discovered and describe a photocatalytic nano-TiO2 phase (anatase) present in the coal ash impacted river water that was also carrying/transporting transition metals (Cu, Fe), which may also have environmental consequences.

Reducing Environmental Toxicity of Silver Nanoparticles through Shape Control.

The use of antibacterial silver nanomaterials in consumer products ranging from textiles to toys has given rise to concerns over their environmental toxicity. These materials, primarily nanoparticles, have been shown to be toxic to a wide range of organisms; thus methods and materials that reduce their environmental toxicity while retaining their useful antibacterial properties can potentially solve this problem. Here we demonstrate that silver nanocubes display a lower toxicity toward the model plant species Lolium multiflorum while showing similar toxicity toward other environmentally relevant and model organisms (Danio rerio and Caenorhabditis elegans) and bacterial species (Esherichia coli, Bacillus cereus, and Pseudomonas aeruginosa) compared to quasi-spherical silver nanoparticles and silver nanowires. More specifically, in the L. multiflorum experiments, the roots of silver nanocube treated plants were 5.3% shorter than the control, while silver nanoparticle treated plant roots were 39.6% shorter than the control. The findings here could assist in the future development of new antibacterial products that cause less environmental toxicity after their intended use.

Speciation Matters: Bioavailability of Silver and Silver Sulfide Nanoparticles to Alfalfa (Medicago sativa).

Terrestrial crops are directly exposed to silver nanoparticles (Ag-NPs) and their environmentally transformed analog silver sulfide nanoparticles (Ag2S-NPs) when wastewater treatment biosolids are applied as fertilizer to agricultural soils. This leads to a need to understand their bioavailability to plants. In the present study, the mechanisms of uptake and distribution of silver in alfalfa (Medicago sativa) were quantified and visualized upon hydroponic exposure to Ag-NPs, Ag2S-NPs, and AgNO3 at 3 mg total Ag/L. Total silver uptake was measured in dried roots and shoots, and the spatial distribution of elements was investigated using transmission electron microscopy (TEM) and synchrotron-based X-ray imaging techniques. Despite large differences in release of Ag(+) ions from the particles, Ag-NPs, Ag2S-NPs, and Ag(+) became associated with plant roots to a similar degree, and exhibited similarly limited (<1%) amounts of translocation of silver into the shoot system. X-ray fluorescence (XRF) mapping revealed differences in the distribution of Ag into roots for each treatment. Silver nanoparticles mainly accumulated in the (columella) border cells and elongation zone, whereas Ag(+) accumulated more uniformly throughout the root. In contrast, Ag2S-NPs remained largely adhered to the root exterior, and the presence of cytoplasmic nano-SixOy aggregates was observed. Exclusively in roots exposed to particulate silver, NPs smaller than the originally dosed NPs were identified by TEM in the cell walls. The apparent accumulation of Ag in the root apoplast determined by XRF, and the presence of small NPs in root cell walls suggests uptake of partially dissolved NPs and translocation along the apoplast.

Emerging contaminant or an old toxin in disguise? Silver nanoparticle impacts on ecosystems.

The use of antimicrobial silver nanoparticles (AgNPs) in consumer-products is rising. Much of these AgNPs are expected to enter the wastewater stream, with up to 10% of that eventually released as effluent into aquatic ecosystems with unknown ecological consequences. We examined AgNP impacts on aquatic ecosystems by comparing the effects of two AgNP sizes (12 and 49 nm) to ionic silver (Ag(+); added as AgNO3), a historically problematic contaminant with known impacts. Using 19 wetland mesocosms, we added Ag to the 360 L aquatic compartment to reach 2.5 mg Ag L(-1). Silver treatments and two coating controls were done in triplicate, and compared to four replicate controls. All three silver treatments were toxic to aquatic plants, leading to a significant release of dissolved organic carbon and chloride following exposure. Simultaneously, dissolved methane concentrations increased forty-fold relative to controls in all three Ag treatments. Despite dramatic toxicity differences observed in lab studies for these three forms of Ag, our results show surprising convergence in the direction, magnitude, and duration of ecosystem-scale impacts for all Ag treatments. Our results suggest that all forms of Ag changed solute chemistry driving transformations of Ag which then altered Ag impacts.

Drought-induced saltwater incursion leads to increased wetland nitrogen export.

Coastal wetlands have the capacity to retain and denitrify large quantities of reactive nitrogen (N), making them important in attenuating increased anthropogenic N flux to coastal ecosystems. The ability of coastal wetlands to retain and transform N is being reduced by wetland losses resulting from land development. Nitrogen retention in coastal wetlands is further threatened by the increasing frequency and spatial extent of saltwater inundation in historically freshwater ecosystems, due to the combined effects of dredging, declining river discharge to coastal areas due to human water use, increased drought frequency, and accelerating sea-level rise. Because saltwater incursion may affect N cycling through multiple mechanisms, the impacts of salinization on coastal freshwater wetland N retention and transformation are not well understood. Here, we show that repeated annual saltwater incursion during late summer droughts in the coastal plain of North Carolina changed N export from organic to inorganic forms and led to a doubling of annual NH(4)(+) export from a 440 hectare former agricultural field undergoing wetland restoration. Soil solution NH(4)(+) concentrations in two mature wetlands also increased with salinization, but the magnitude of increase was smaller than that in the former agricultural field. Long-term saltwater exposure experiments with intact soil columns demonstrated that much of the increase in reactive N released could be explained by exchange of salt cations with sediment NH(4)(+). Using these findings together with the predicted flooding of 1661 km(2) of wetlands along the NC coast by 2100, we estimate that saltwater incursion into these coastal areas could release up to 18 077 Mg N, or approximately half the annual NH(4)(+) flux of the Mississippi River. Our results suggest that saltwater incursion into coastal freshwater wetlands globally could lead to increased N loading to sensitive coastal waters.

Sulfidation of silver nanoparticles: natural antidote to their toxicity.

Nanomaterials are highly dynamic in biological and environmental media. A critical need for advancing environmental health and safety research for nanomaterials is to identify physical and chemical transformations that affect the nanomaterial properties and their toxicity. Silver nanoparticles, one of the most toxic and well-studied nanomaterials, readily react with sulfide to form Ag(0)/Ag2S core-shell particles. Here, we show that sulfidation decreased silver nanoparticle toxicity to four diverse types of aquatic and terrestrial eukaryotic organisms (Danio rerio (zebrafish), Fundulus heteroclitus (killifish), Caenorhabditis elegans (nematode worm), and the aquatic plant Lemna minuta (least duckweed)). Toxicity reduction, which was dramatic in killifish and duckweed even for low extents of sulfidation (about 2 mol % S), is primarily associated with a decrease in Ag(+) concentration after sulfidation due to the lower solubility of Ag2S relative to elemental Ag (Ag(0)). These results suggest that even partial sulfidation of AgNP will decrease the toxicity of AgNPs relative to their pristine counterparts. We also show that, for a given organism, the presence of chloride in the exposure media strongly affects the toxicity results by affecting Ag speciation. These results highlight the need to consider environmental transformations of NPs in assessing their toxicity to accurately portray their potential environmental risks.

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.

Biotic and abiotic interactions in aquatic microcosms determine fate and toxicity of Ag nanoparticles: part 2-toxicity and Ag speciation.

To study the effects of complex environmental media on silver nanoparticle (AgNP) toxicity, AgNPs were added to microcosms with freshwater sediments and two species of aquatic plants (Potamogeton diversifolius and Egeria densa), followed by toxicity testing with microcosm surface water. Microcosms were designed with four environmental matrices in order to determine the contribution of each environmental compartment to changes in toxicity: water only (W), water + sediment (WS), water + plants (WP), and water + plants + sediment (WPS). Silver treatments included AgNPs with two different coatings, gum arabic (GA-AgNPs) or polyvinylpyrollidone (PVP-AgNPs), as well as AgNO(3). Water samples taken from the microcosms at 24 h postdosing were used in acute toxicity tests with two standard model organisms, early life stage zebrafish (Danio rerio) and Daphnia magna. Speciation of Ag in these samples was analyzed using Ag L3-edge X-ray absorption near edge spectroscopy (XANES). Silver speciation patterns for the nanoparticle treatments varied significantly by coating type. While PVP-AgNPs were quite stable and resisted transformation across all matrices (>92.4% Ag(0)), GA-AgNP speciation patterns suggest significantly higher transformation rates, especially in treatments with plants (<69.2% and <58.8% Ag(0) in WP and WPS, respectively) and moderately increased transformation with sediments (<85.6% Ag(0)). Additionally, the presence of plants in the microcosms (with and without sediments) reduced both the concentration of Ag in the water column and toxicity for all Ag treatments. Reductions in toxicity may have been related to decreased water column concentrations as well as changes in the surface chemistry of the particles induced by organic substances released from the plants.

Long-term transformation and fate of manufactured ag nanoparticles in a simulated large scale freshwater emergent wetland.

Transformations and long-term fate of engineered nanomaterials must be measured in realistic complex natural systems to accurately assess the risks that they may pose. Here, we determine the long-term behavior of poly(vinylpyrrolidone)-coated silver nanoparticles (AgNPs) in freshwater mesocosms simulating an emergent wetland environment. AgNPs were either applied to the water column or to the terrestrial soils. The distribution of silver among water, solids, and biota, and Ag speciation in soils and sediment was determined 18 months after dosing. Most (70 wt %) of the added Ag resided in the soils and sediments, and largely remained in the compartment in which they were dosed. However, some movement between soil and sediment was observed. Movement of AgNPs from terrestrial soils to sediments was more facile than from sediments to soils, suggesting that erosion and runoff is a potential pathway for AgNPs to enter waterways. The AgNPs in terrestrial soils were transformed to Ag(2)S (~52%), whereas AgNPs in the subaquatic sediment were present as Ag(2)S (55%) and Ag-sulfhydryl compounds (27%). Despite significant sulfidation of the AgNPs, a fraction of the added Ag resided in the terrestrial plant biomass (~3 wt % for the terrestrially dosed mesocosm), and relatively high body burdens of Ag (0.5-3.3 µg Ag/g wet weight) were found in mosquito fish and chironomids in both mesocosms. Thus, Ag from the NPs remained bioavailable even after partial sulfidation and when water column total Ag concentrations are low (<0.002 mg/L).

Antimicrobial effects of commercial silver nanoparticles are attenuated in natural streamwater and sediment.

Given the demonstrated antimicrobial properties of silver nanoparticles (AgNPs), and the key role that microorganisms play in performing critical ecosystem functions such as decomposition and nutrient cycling, there is growing concern that AgNP pollution may negatively impact ecosystems. We examined the response of streamwater and sediment microorganisms to commercially available 21 ± 17 nm AgNPs, and compared AgNP impacts to those of dissolved-Ag added as AgNO(3). We show that in streamwater, AgNPs and AgNO(3) decreased respiration in proportion to dissolved-Ag concentrations at the end of the incubation (r(2) = 0.78), while in sediment the only measurable effect of AgNPs was a 14 % decrease in sulfate concentration. This contrasts with the stronger effects of dissolved-Ag additions in both streamwater and sediment. In streamwater, addition of dissolved-Ag at a level equivalent to the lowest AgNP dose led to respiration below detection, a 55 % drop in phosphatase enzyme activity, and a 10-fold increase in phosphate concentration. In sediment, AgNO(3) addition at a level equivalent to the highest AgNP addition led to a 34 % decrease in respiration, a 55 % increase in microbial biomass, and a shift in bacterial community composition. The results of this study suggest that, in similar freshwater environments, the short-term biological impacts of AgNPs on microbes are attenuated by the physical and chemical properties of streamwater and sediment.

Characterization and environmental implications of nano- and larger TiO(2) particles in sewage sludge, and soils amended with sewage sludge.

Titanium dioxide (TiO(2)) is the most extensively used engineered nanoparticle to date, yet its fate in the soil environment has been investigated only rarely and is poorly understood. In the present study, we conducted two field-scale investigations to better describe TiO(2) nano- and larger particles in their most likely route of entry into the environment, i.e., the application of biosolids to soils. We particularly concentrated on the particles in the nano-size regime due to their novel and commercially useful properties. First, we analyzed three sewage sludge products from the US EPA TNSSS sampling inventory for the occurrence, qualitative abundance, and nature of TiO(2) nano- and larger particles by using analytical scanning electron microscopy and analytical (scanning) transmission electron microscopy. Nano- and larger particles of TiO(2) were repeatedly identified across the sewage sludge types tested, providing strong evidence of their likely concentration in sewage sludge products. The TiO(2) particles identified were as small as 40 nm, and as large as 300 nm, having faceted shapes with the rutile crystal structure, and they typically formed small, loosely packed aggregates. Second, we examined surface soils in mesocosms that had been amended with Ag nanoparticle-spiked biosolids for the occurrence of TiO(2) particles. An aggregate of TiO(2) nanoparticles with the rutile structure was again identified, but this time TiO(2) nanoparticles were found to contain Ag on their surfaces. This suggests that TiO(2) nanoparticles from biosolids can interact with toxic trace metals that would then enter the environment as a soil amendment. Therefore, the long-term behavior of TiO(2) nano- and larger particles in sewage sludge materials as well as their impacts in the soil environment need to be carefully considered.

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.