Comparative phototoxicity of nanoparticulate and bulk ZnO to a free-living nematode Caenorhabditis elegans: the importance of illumination mode and primary particle size.

The present study evaluated phototoxicity of nanoparticulate ZnO and bulk-ZnO under natural sunlight (NSL) versus ambient artificial laboratory light (AALL) illumination to a free-living nematode Caenorhabditis elegans. Phototoxicity of nano-ZnO and bulk-ZnO was largely dependent on illumination method as 2-h exposure under NSL caused significantly greater mortality in C. elegans than under AALL. This phototoxicity was closely related to photocatalytic reactive oxygen species (ROS) generation by the ZnO particles as indicated by concomitant methylene blue photodegradation. Both materials caused mortality in C. elegans under AALL during 24-h exposure although neither degraded methylene blue, suggesting mechanisms of toxicity other than photocatalytic ROS generation were involved. Particle dissolution of ZnO did not appear to play an important role in the toxicity observed in this study. Nano-ZnO showed greater phototoxicity than bulk-ZnO despite their similar size of aggregates, suggesting primary particle size is more important than aggregate size in determining phototoxicity.

Fate of arsenic compounds in poultry litter upon land application.

The use of the organic As compound, roxarsone, as an antibiotic additive to poultry feed continues to raise concern over potential negative environmental impacts. Total As concentration in poultry litter can reach >40 mg kg(-1) and both roxarsone and its mineralization product As(V) have been identified in poultry litters (PL). To investigate the fate of these As species upon land application of PL we conducted two studies. In the first, an Orangeburg soil (Ultisol from the Atlantic Coastal Plain) was spiked with either 20 mg kg(-1) As(V) or roxarsone and incubated at 10% moisture content for 4 months. Exchangeable As was determined periodically by extraction with 0.1M PO(4). Both As(V) and roxarsone displayed similar desorption; initially, approximately 70% of added As was ligand exchangeable and this decreased to 35% after 4 months incubation, presumably due to either slow sorption reactions or a change in solid phase speciation of As to less exchangeable forms. In the second study, various manipulations of two PL samples were applied to the Orangeburg soil at realistic field application rates. The treatments were wet to 10% moisture content and water soluble As, Cu and organic carbon (DOC) was measured over 30 days. Arsenic and Cu solubility were highest from the dried litter samples. Ashing of the PLs decreased soluble As and Cu, presumably because of the loss of organic matter from the ashed litter and subsequent decrease in DOC. Application of leachates from either PL resulted in higher concentrations of soluble As and Cu than when the soil was amended with equivalent concentrations of soluble As and Cu dissolved in DI H(2)O. We hypothesize that the increased levels of DOC from the PL treatments enhance As and Cu solubility through competitive sorption and complexation, respectively. In fact, As and Cu solubility was correlated to DOC levels in the amended soil extracts. Even though land application of PL introduced relatively low concentrations of As and Cu to soil it appeared that other soluble constituents of PL significantly enhanced As and Cu solubility.

Evidence for biogenic pyromorphite formation by the nematode Caenorhabditis elegans.

The determination of chemical speciation and spatial distribution is a prerequisite for a mechanistic understanding of contaminant bioavailability and toxicity to an organism. We have employed synchrotron X-ray techniques to study Cu and Pb speciation and spatial distribution in the soil nematode Caenorhabditis elegans. Nematodes were exposed to each metal ion singly or simultaneously in solution for 24 h and were then rinsed thoroughly and preserved in formalin for transportation to the National Synchrotron Light Source. Experiments were conducted at the microprobe beamline X26A employing a focused beam of approximately 10 microm in diameter. Nematodes were mounted in agar gel on Kapton tape. Two-dimensional elemental maps for Cu- and Pb-exposed nematodes were collected in fluorescence mode. Copper was homogeneously distributed throughout the body of the nematode, but Pb exhibited a high degree of localization in the nematode, exclusively in the anterior pharynx region. Detectable localized concentrations of Pb in C. elegans occurred at aqueous exposure concentrations of 2.4 microM. Micro X-ray diffraction of these Pb hotspots gave a diffraction pattern indicating a crystalline Pb solid that was consistent with the Pb phosphate, pyromorphite. Biogenic inorganic phosphate granule formation is relatively common in soil invertebrates; however, these phosphates are typically amorphous, and we believe that this is the first report of crystalline pyromorphite formed internally in an organism.

Trace element speciation in poultry litter.

Trace elements are added to poultry feed for disease prevention and enhanced feed efficiency. High concentrations are found in poultry litter (PL), which raises concerns regarding trace element loading of soils. Trace metal cation solubility from PL may be enhanced by complexation with dissolved organic carbon (DOC). Mineralization of organo-As compounds may result in more toxic species such as As(III) and As(V). Speciation of these elements in PL leachates should assist in predicting their fate in soil. Elemental concentrations of 40 PL samples from the southeastern USA were determined. Water-soluble extractions (WSE) were fractionated into hydrophobic, anionic, and cationic species with solid-phase extraction columns. Arsenic speciation of seven As species, including the main As poultry feed additives, roxarsone (ROX; 3-nitro-4-hydroxyphenylarsonic acid) and p-arsanilic acid (p-ASA; 4-aminophenylarsonic acid), was performed by ion chromatography-inductively coupled plasma-mass spectrometry (IC-ICP-MS). Total As concentrations in the litter varied from 1 to 39 mg kg(-1), averaging 16 mg kg(-1). Mean total Cu, Ni, and Zn concentrations were 479, 11, and 373 mg kg(-1), respectively. Copper and Ni were relatively soluble (49 and 41% respectively) while only 6% of Zn was soluble. Arsenic was highly soluble with an average of 71% WSE. Roxarsone was the major As species in 50% of PL samples. However, the presence of As(V) as the major species in 50% of the PL samples indicates that mineralization of ROX had occurred. The high solubility of As from litter and its apparent ready mineralization to inorganic forms coupled with the large quantity of litter that is annually land-applied in the USA suggests a potential detrimental effect on soil and water quality in the long term.

Trace element speciation in largemouth bass ( Micropterus salmoides) from a fly ash settling basin by liquid chromatography-ICP-MS.

Analytical techniques used to examine the chemical speciation of multiple trace elements are important for the investigation of biological systems. Size exclusion chromatography (SEC) coupled to ICP-MS was used to investigate the speciation of Se, As, Cu, Cd and Zn in tissue extracts from a largemouth bass (Micropterus salmoides) collected from a coal fly ash basin and results were compared to a largemouth bass collected at a reference site. Using a Biosil SEC column, with an effective separation range of 100-7 KDa, Cu, Zn and Cd were shown to be bound to metallothionein (MT) in the liver, gill and, to a lesser extent, gonad tissue extract. In liver, muscle and gill of the ash basin bass, Se was predominantly present as low molecular weight species. Only in the gonad extract was the major fraction of Se associated with high molecular weight species. For the liver and gill extracts, further SEC-ICP-MS on a column with an effective separation range of 7000-500 Da was performed, but Se species still eluted near the total volume of the column suggesting a low molecular weight organic or inorganic species. Ion chromatography (IC)-ICP-MS using an AS7 column and HNO(3) gradient elution indicated that the Se and As species in the liver and gill extracts had similar retention times but these retention times did not correspond to retention times for As(III), As(V), dimethylarsenate, arsenobetaine, Se(IV), Se(VI), seleno-methionine, or seleno-cystine.

Immobilization of cesium-137 and uranium in contaminated sediments using soil amendments.

Batch and dynamic leaching methods were used to evaluate the effectiveness of hydroxyapatite (HA), illite, and zeolite, alone and in combination, as soil additives for reducing the migration of cesium-137 (137Cs+) and uranium (U) from contaminated sediments. Amendment treatments ranging from 0 to 50 g kg(-1) were added to the sediment and equilibrated in 0.001 M CaCl2. After equilibration, the treatment supernatants were analyzed for 137Cs+, U, PO4, and other metals. The residual sediments were then extracted overnight using one of the following: 1.0 M NH4Cl, 0.5 M CaCl2, or the Toxicity Characteristic Leaching Procedure (TCLP) extractant. Cesium was strongly sorbed to the contaminated sediments, presumably due to interlayer fixation within native illitic clays. In fact, 137Cs+ was below detection limits in the initial equilibration solutions, the CaCl2 extract, and the TCLP solution, regardless of amendment. Extractants selective for interlayer cations (1.0 M NH4Cl) were necessary to extract measurable levels of 137Cs+. Addition of illitic clays further reduced Cs+ extractability, even when subjected to the aggressive extractants. Zeolite, however, was ineffective in reducing Cs+ mobility when subjected to the aggressive extractants. Hydroxyapatite was less effective than illite at reducing NH4+-extractable Cs+. Hydroxyapatite, and mixtures of HA with illite or zeolite, were highly effective in reducing U extractability in both batch and leaching tests. Uranium immobilization by HA was rapid with similar final U concentrations observed for equilibration times ranging from 1 h to 30 d. The current results demonstrate the effectiveness of soil amendments in reducing the mobility of U and Cs+, which makes in-place immobilization an effective remediation alternative.

Comparison of two micro-analytical methods for detecting the spatial distribution of sorbed Pu on geologic materials.

Subsurface transport of groundwater contaminants is greatly influenced by chemical speciation, precipitation and sorption processes at the mineral-water interface. The retardation of contaminants is often greatest at boundaries between minerals and in fractures and pore spaces. The investigation of the spatial distribution of sorbed contaminants along these boundaries requires micro-analytical techniques. The sorption of dissolved Pu(V) on a natural zeolitic tuff from Yucca Mountain (NV, USA) was examined using microautoradiography (MAR), X-ray diffraction (XRD), electron microprobe (EM) techniques, and synchrotron-based micro-X-ray fluorescence (micro-SXRF). The tuff contained a heterogeneous distribution of zeolites and trace quantities of smectites, Fe oxides (hematite), and Mn oxides (rancieite), which are present as fracture fill and pore space materials. Micro-SXRF studies showed that Pu is mostly associated with bodies of smectite plus Mn oxides, which were typically elevated in Ce, Ga, Nb, Pb, Y, Ca, Ti, and Zn. Sorbed Pu was not associated with Fe-rich bodies, which were enriched in Cl and Rb. Results of the MAR studies were complementary to that of the micro-SXRF studies in that Pu was associated with similar elements in the tuff. Indirect detection of Pu by EM or micro-SXRF (by analyzing Ag developed on the MAR photoemulsion) was a more sensitive method for detecting lower levels of sorbed Pu than the direct detection of sorbed Pu via micro-SXRF in the absence of the photoemulsion.

Immobilization of nickel and other metals in contaminated sediments by hydroxyapatite addition.

Batch experiments were conducted to evaluate the ability of hydroxyapatte (HA) to reduce the solubility of metals, including the primary contaminants of concern, Ni and U, from contaminated sediments located on the Department of Energy's Savannah River Site, near Aiken, SC. Hydroxyapatitie was added to the sediments at application rates of 0, 5, 15.8, and 50 g kg-1. After equilibrating in either 0.02 M KCl or 0.01 M CaCl2, the samples were centrifuged and the supernatants filtered prior to metal, dissolved organic C, and PO4 analyses. The treated soils were then air-dried and changes in solid-phase metal distribution were evaluated using sequential extractions and electron-based microanalysis techniques. Hydroxyapatite was effective at reducing the solubility of U and, to a lesser degree, Ni. Hydroxyapatite was also effective in reducing the solubility of Al, Ba, Cd, Co, Mn, and Pb. Sequential extractions indicate that HA transfers such metals from more chemically labile forms, such as the water-soluble and exchangeable fractions, by altering solid-phase speciation in favor of secondary phosphate precipitates. Hydroxyapatite effectiveness was somewhat reduced in the presence of soluble organics that likely increased contaminant metal solubility through complexation. Arsenic and Cr solubility increased with HA addition, suggesting that the increase in pH and competition from PO4 reduced sorption of oxyanion contaminants. Energy dispersive x-ray (EDXA) analysis conducted in the transmission electron microscope (TEM) confirmed that HA amendment sequesters U, Ni, Pb, and possibly other contaminant metals in association with secondary Al-phosphates.

Determination of arsenic speciation in poultry wastes by IC-ICP-MS.

The aromatic organoarsenic compounds 4-hydroxy 3-nitrobenzenearsenic acid (Roxarsone, ROX) or 4-aminoben-zenearsenic acid (p-arsanilic acid, p-ASA) are used as feed additives in the poultry industry for disease control and enhanced feed efficiency. While federal regulations govern acceptable As concentrations in edible tissue, elevated As concentrations occur in poultry litter, which raises concerns over repeated land application of poultry litter in agriculture. As a precursor to studying the fate of these organoarsenic compounds in soils, three speciation methodologies were developed to separate ROX and p-ASA from the more common and more toxic As species arsenate, arsenite, dimethyl arsenic acid (DMA), and monomethyl arsenic acid (MMA). The six arsenic species were separated on a Dionex AS14 column using a PO4 eluant, an AS16 column using a OH- eluant, and an AS7 column using a HNO3 eluant. While all three methods provided detection limits below 0.5 microg L(-1) for all species, detection limits were lowest for the AS16 and AS7 columns, where all detection limits were generally < 0.05 microg L(-1). The major arsenic species in a water extract of a poultry litter sample was identified as ROX by all three methods with trace concentrations of DMA and As(V) also detected. The AS14 and AS16 separations also revealed a number of unidentified As species present at low concentrations, presumably metabolites of ROX. This methodology should prove useful in identifying organoarsenic compounds and the more toxic inorganic species in soils subject to poultry litter application.

Direct measurement of aluminum uptake and distribution in single cells of Chara corallina.

Quantitative information on the uptake and distribution of Al at the cellular level is required to understand mechanisms of Al toxicity, but direct measurement of uptake across the plasma membrane has remained elusive. We measured rates of Al transport across membranes in single cells of Chara corallina using the rare (26)Al isotope, an emerging technology (accelerator mass spectrometry), and a surgical technique for isolating subcellular compartments. Accumulation of Al in the cell wall dominated total uptake (71-318 microgram m(-2) min(-1)), although transport across the plasma membrane was detectable (71-540 ng m(-2) min(-1)) within 30 min of exposure. Transport across the tonoplast was initially negligible, but accelerated to rates approximating uptake across the plasma membrane. The avacuolate protoplasm showed signs of saturation after 60 min, but continued movement across the plasma membrane was supported by sequestration in the vacuole. Saturation of all compartments was observed after 12 to 24 h. Accumulation of Al in the cell wall reflected variation in [Al(3+)] induced by changes in Al supply or complexing ligands, but was unaffected by pH. In contrast, transport across the plasma membrane peaked at pH 4.3 and increased when [Al(3+)] was reduced by complexing ligands. Cold temperature (4 degrees C) reduced accumulation in the cell wall and protoplasm, whereas 2,4-dinitrophenol and m-chlorocarbonylcyanidephenyl hydrazone increased membrane transport by 12- to 13-fold. Our data suggest that the cell wall is the major site of Al accumulation. Nonetheless, membrane transport occurs within minutes of exposure and is supported by subsequent sequestration in the vacuole. The rapid delivery of Al to the protoplasm suggests that intracellular lesions may be possible.

Characterization of complex mineral assemblages: implications for contaminant transport and environmental remediation.

Surface reactive phases of soils and aquifers, comprised of phyllosilicate and metal oxohydroxide minerals along with humic substances, play a critical role in the regulation of contaminant fate and transport. Much of our knowledge concerning contaminant-mineral interactions at the molecular level, however, is derived from extensive experimentation on model mineral systems. Although these investigations have provided a foundation for understanding reactive surface functional groups on individual mineral phases, the information cannot be readily extrapolated to complex mineral assemblages in natural systems. Recent studies have elucidated the role of less abundant mineral and organic substrates as important surface chemical modifiers and have demonstrated complex coupling of reactivity between permanent-charge phyllosilicates and variable-charge Fe-oxohydroxide phases. Surface chemical modifiers were observed to control colloid generation and transport processes in surface and subsurface environments as well as the transport of solutes and ionic tracers. The surface charging mechanisms operative in the complex mineral assemblages cannot be predicted based on bulk mineralogy or by considering surface reactivity of less abundant mineral phases based on results from model systems. The fragile nature of mineral assemblages isolated from natural systems requires novel techniques and experimental approaches for investigating their surface chemistry and reactivity free of artifacts. A complete understanding of the surface chemistry of complex mineral assemblages is prerequisite to accurately assessing environmental and human health risks of contaminants or in designing environmentally sound, cost-effective chemical and biological remediation strategies.

Immobilization of Uranium in Contaminated Sediments by Hydroxyapatite Addition.

Batch equilibrations were performed to investigate the ability of hydroxyapatite (Ca5(PO4)3OH) to chemically immobilize U in two contaminated sediment samples having different organic carbon contents (123 and 49 g kg(-1), respectively). Apatite additions lowered aqueous U to near proposed drinking water standards in batch equilibrations of two distinct sediment strata having total U concentrations of 1703 and 2100 mg kg(-1), respectively. Apatite addition of 50 g kg(-1) reduced the solubility of U to values less than would be expected if autunite (Ca(UO2)2(PO4)2·10H2O) was the controlling solid phase. A comparison of the two sediment types suggests that aqueous phase U may be controlled by both the DOC content through complexation and the equilibrium pH for a given apatite application rate. Sequential chemical extractions demonstrated that apatite amendment transfers U from more chemically labile fractions, including water-soluble, exchangeable, and acid-soluble (pH ≈ 2.55) fractions, to the Mn-occluded fraction (pH ≈ 1.26). This suggests that apatite amendment redirects solid-phase speciation with secondary U phosphates being solubilized due to the lower pH of the Mn-occluded extractant, despite the lack of significant quantities of Mn oxides within these sediments. Energy dispersive X-ray (EDX) analysis conducted in a transmission electron microscope (TEM) confirmed that apatite amendment sequesters some U in secondary Al/Fe phosphate phases.

The hydrolytic products of aluminum and their biological significance.

The relative distribution of Al between its various organic and inorganic complexes dictates its mobility in the environment, bioavailability, and toxicity. In recent years, there has been significant progress made in understanding the differential bioavailability and toxicity of various chemical species of Al to plants and certain aquatic organisms. Far less information concerning chemical speciation and differential uptake and transport of Al in humans is available. Among the important inorganic complexes of interest are the hydrolyzed-Al species, particularly the nonequilibrium, metastable polynuclear complexes, which form readily, have a fairly wide stability range, and have been demonstrated toxic to plants and fish. In recent years(27)Al NMR spectroscopy has provided significant direct information on the polynuclear complexes existing in a wide range of aqueous solutions. The [Al12O4(OH)24+n(H2O)12-n]((7-n)+) polynuclear complex is often found to be the predominant species in partially neutralized Al solutions and has recently been demonstrated to be more toxic to certain plants than the hexaaqua Al cation. It is also the principal component of Al-chlorohydrate, a highly soluble antiperspirant, present in many hydrolyzed Al solutions utilized in water and waste water treatment, and, as hypothesized herein, a primary constituent of many hydroxide gels utilized as antacids. This polynuclear has a wide pH stability range, reportedly forms copolynuclears with Si, and contains tetrahedrally coordinated Al within its structure, all features that may be relevant to the recently reported properties of Al associated with neuritic plaque cores.