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Pteris vittata is the first-reported arsenic (As) hyperaccumulator, which has been applied to phytoremediation of As-contaminated soil. PvACR3, a key arsenite (AsIII) antiporter, plays an important role in As hyperaccumulation in P. vittata . However, its functions in plants are not fully understood. In this study, the PvACR3 gene was heterologously expressed in tobacco, driven by its native promoter ( Pro PvACR3 ). After growing at 5 μM AsIII or 10 μM AsV in hydroponics for 1-5 days, PvACR3 -expression enhanced the As levels in leaves by 66.4-113 and 51.8-101%, without impacting the As contents in the roots or stems. When cultivated in As-contaminated soil, PvACR3 -expressed transgenic plants accumulated 47.9-85.5% greater As in the leaves than wild-type plants. In addition, PvACR3 -expression increased the As resistance in transgenic tobacco, showing that enhanced leaf As levels are not detrimental to its overall As tolerance. PvACR3 was mainly expressed in tobacco leaf veins and was likely to unload AsIII from the vein xylem vessels to the mesophyll cells, thus elevating the leaf As levels. This work demonstrates that heterologously expressing PvACR3 under its native promoter specifically enhances leaf As accumulation in tobacco, which helps to reveal the As-hyperaccumulation mechanism in P. vittata and to enhance the As accumulation in plant leaves for phytoremediation.

Sparingly-soluble phosphate rock (PR), a raw material for P-fertilizer production, can be effectively utilized by the As-hyperaccumulator Pteris vittata but not most plants. In this study, we investigated the associated mechanisms by measuring dissolved organic carbon (DOC) and acid phosphatase in the rhizosphere, and nutrient uptake and gene expression related to the As metabolism in P. vittata . The plants were grown in a soil containing 200 mg kg -1 As and/or 1.5% PR for 30 days. Compared to the As treatment, the P. vittata biomass was increased by 33% to 4.6 g plant -1 in the As+PR treatment, corresponding to 27% decrease in its frond oxidative stress as measured by malondialdehyde. Due to PR-enhanced DOC production in the rhizosphere, the Ca, P, and As contents in P. vittata fronds were increased by 17% to 9.7 g kg -1 , 29% to 5.0 g kg -1 , and 57% to 1045 mg kg -1 in the As+PR treatment, thereby supporting its better growth. Besides, PR-induced rhizosphere pH increase from 5.0 to 6.9 promoted greater P uptake by P. vittata probably via upregulating low-affinity P transporters PvPTB1;1/1;2 by 3.7-4.1 folds. Consequently, 29% lower available-P induced the 3.3-fold upregulation of high-affinity P transporter PvPht1;3 in the As+PR treatment, which was probably responsible for the 58% decrease in available-As content in the rhizosphere. Consistent with the enhanced As translocation and sequestration, arsenite antiporters PvACR3/3;3 were upregulated by 1.8-4.4 folds in the As+PR than As treatment. In short, sparingly-soluble PR enhanced the Ca, P, and As availability in P. vittata rhizosphere and improved their uptake via upregulating genes related to As metabolism, suggesting its potential application for improving phytoremediation in As-contaminated soils.

Arsenic (As) contamination in soil poses a potential threat to human health via crop uptake. As-hyperaccumulator Pteris vittata serves as a model plant to study As uptake and associated mechanisms. This study focuses on a novel P/AsV transport system mediated by low-affinity phosphate transporter-B 1 family (PTB1) in P. vittata . Here, we identified two plasma-membrane-localized PTB1 genes, PvPTB1;1/1;2 , in vascular plants for the first time, which were 4.4-40-fold greater in expression in P. vittata than in other Pteris ferns. Functional complementation of a yeast P-uptake mutant and enhanced P accumulation in transgenic Arabidopsis thaliana confirmed their role in P uptake. Moreover, the expression of PvPTB1;1/1;2 facilitated the transport and accumulation of As in both yeast and A. thaliana shoots, demonstrating a comparable AsV uptake capacity. Microdissection-qPCR analysis and single-cell transcriptome analysis collectively suggest that PvPTB1;1/1;2 are specifically expressed in the epidermal cells of P. vittata roots. PTB1 may play a pivotal role in efficient P recycling during phytate secretion and hydrolysis in P. vittata roots. In summary, the dual P transport mechanisms consisting of high-affinity Pht1 and low-affinity PTB1 may have contributed to the efficient P/As uptake in P. vittata , thereby contributing to efficient phytoremediation for As-contaminated soils.

Phytate, the principal P storage in plant seeds, is also an important organic P in soils, but it is unavailable for plant uptake. However, the As-hyperaccumulator Pteris vittata can effectively utilize soluble Na-phytate, while its ability to utilize insoluble Ca/Fe-phytate is unclear. Here, we investigated phytate uptake and the underlying mechanisms based on the phytase activity, nutrient uptake, and expression of genes involved in As metabolisms. P. vittata plants were cultivated hydroponically in 0.2-strength Hoagland nutrient solution containing 50 μM As and 0.2 mM Na/Ca/Fe-phytate, with 0.2 mM soluble-P as the control. As the sole P source, all three phytates supported P. vittata growth, with its biomass being 3.2-4.1 g plant -1 and Ca/Fe-phytate being 19-29% more effective than Na-phytate. Phytate supplied soluble P to P. vittata probably via phytase hydrolysis, which was supported by 0.4-0.7 nmol P min -1 g -1 root fresh weight day -1 phytase activity in its root exudates, with 29-545 μM phytate-P being released into the growth media. Besides, compared to Na-phytate, Ca/Fe-phytate enhanced the As contents by 102-140% to 657-781 mg kg -1 in P. vittata roots and by 43-86% to 1109-1447 mg kg -1 in the fronds, which was accompanied by 21-108% increase in Ca and Fe uptake. The increased plant As is probably attributed to 1.3-2.6 fold upregulation of P transporters PvPht1;3/4 for root As uptake, and 1.8-4.3 fold upregulation of arsenite antiporters PvACR3/3;1/3;3 for As translocation to and As sequestration into the fronds. This is the first report to show that, besides soluble Na-phytate, P. vittata can also effectively utilize insoluble Ca/Fe-phytate as the sole P source, which sheds light onto improving its application in phytoremediation of As-contaminated sites.

Prebiotics may stimulate beneficial gut microorganisms. However, it remains unclear whether they can lower the oral bioavailability of early life arsenic (As) exposure via regulating gut microbiota and altering As biotransformation along the gastrointestinal (GI) tract. In this study, weanling mice were exposed to arsenate (iAs V ) via diet (7.5 μg As g -1 ) amended with fructooligosaccharides (FOS), galactooligosaccharides (GOS), and inulin individually at 1% and 5% (w/w). Compared to As exposure control mice, As concentrations in mouse blood, liver, and kidneys and As urinary excretion factor (UEF) were reduced by 43.7%-74.1% when treated with 5% GOS. The decrease corresponded to a significant proliferation of Akkermansia and Psychrobacter , reduced percentage of inorganic arsenite (iAs III ) and iAs V by 47.4% and 65.4%, and increased proportion of DMA V in intestinal contents by 101% in the guts of mice treated with 5% GOS compared to the As control group. In contrast, FOS and inulin either at l% or 5% did not reduce As concentration in mouse blood, liver, and kidneys or As UEF. These results suggest that GOS supplementation may be a gut microbiota-regulating approach to lower early life As exposure via stimulating the growth of Akkermansia and Psychrobacter and enhancing As methylation in the GI tract.

Few studies have investigated the long-term effect of exposure to arsenic (As), lead (Pb), and cadmium (Cd) via drinking water at the provisional guideline values on gut microflora. In this study, male and female mice were exposed to water As, Pb, or Cd at 10, 10, or 5 μg L -1 for 6 months. At the end of the exposure, the net weight gain of male mice exposed to As and Pb (9.91 ± 1.35 and 11.2 ± 1.50 g) was significantly ( p < 0.05) lower compared to unexposed control mice (14.1 ± 3.24 g), while this was not observed for female mice. Relative abundance of Akkermansia , a protective gut bacterium against intestinal inflammation, was reduced from 29.7% to 3.20%, 4.83%, and 17.0% after As, Pb, and Cd exposure in male mice, which likely caused chronic intestinal inflammation, as suggested by 2.81- to 9.60-fold higher mRNA levels of pro-inflammatory factors in ileal enterocytes of male mice. These results indicate that long-term exposure to drinking water As, Pb, and Cd at concentrations equivalent to the China provisional guideline values can cause loss of protective bacteria and lead to chronic intestinal inflammation, thereby affecting body weight gain in male mice.

Rare earth elements (REEs) are critical for numerous modern technologies, and demand is increasing globally; however, production steps are resource-intensive and environmentally damaging. Some plant species are able to hyperaccumulate REEs, and understanding the biology behind this phenomenon could play a pivotal role in developing more environmentally friendly REE recovery technologies. Here, we identified a REE transporter NRAMP REE Transporter 1 (NREET1) from the REE hyperaccumulator fern Dicranopteris linearis . Although NREET1 belongs to the natural resistance-associated macrophage protein (NRAMP) family, it shares a low similarity with other NRAMP members. When expressed in yeast, NREET1 exhibited REE transport capacity, but it could not transport divalent metals, such as zinc, nickel, manganese, or iron. NREET1 is mainly expressed in D. linearis roots and predominantly localized in the plasma membrane. Expression studies in Arabidopsis thaliana revealed that NREET1 functions as a transporter mediating REE uptake and transfer from root cell walls into the cytoplasm. Moreover, NREET1 has a higher affinity for transporting light REEs compared to heavy REEs, which is consistent to the preferential enrichment of light REEs in field-grown D. linearis . We therefore conclude that NREET1 may play an important role in the uptake and consequently hyperaccumulation of REEs in D. linearis . These findings lay the foundation for the use of synthetic biology techniques to design and produce sustainable, plant-based REE recovery systems.

Early-life arsenic (As) exposure is a particular health concern. However, it is unknown if As ingested early in life is more readily absorbed from the gastrointestinal (GI) tract, i.e., higher in oral bioavailability. Here, weanling (3-week) and adult (6-week-old) female mice were exposed to arsenate in the diet (10 μg g -1 ) over a 3-week period with As oral bioavailability estimated using As urinary excretion as the bioavailability endpoint. The As urinary excretion factor was 1.54-fold higher in weanling mice compared to adult mice (82.2 ± 7.29 versus 53.1 ± 3.73%), while weanling mice also showed 2.28-, 1.50-, 1.48-, and 1.89-fold higher As concentration in small intestine tissue, blood, liver, and kidneys, demonstrating significantly higher As oral bioavailability of early-life exposure. Compared to adult mice, weanling mice significantly differed in gut microbiota, but the difference did not lead to remarkable differences in As biotransformation in the GI tract or tissue and in overall gut metabolite composition. Although the expression of several metabolites (e.g., atrolactic acid, hydroxyphenyllactic acid, and xanthine) was up-regulated in weanling mice, they had limited ability to elevate As solubility in the intestinal tract. Compared to adult mice, the intestinal barrier function and intestinal expression of phosphate transporters responsible for arsenate absorption were similar in weanling mice. However, the small intestine of weanling mice was characterized by more defined intestinal villi with greater length and smaller width, providing a greater surface area for As to be absorbed across the GI barrier. The results highlight that early-life As exposure can be more readily absorbed, advancing the understanding of its health risk.

Phytate as a root exudate is rare in plants as it mainly serves as a P storage in the seeds; however, As-hyperaccumulator Pteris vittata effectively secretes phytate and utilizes phytate-P, especially under As exposure. This study investigated the effects of As on its phytate and phytase exudation and the impacts of As and/or phytate on each other's uptake in P. vittata through two hydroponic experiments. Under 10-100 μM arsenate (AsV), the exudation of phytate and phytase by P. vittata was increased by 50-72% to 20.4-23.4 μmol h -1 g -1 and by 28-104% to 18.6-29.5 nmol h -1 plant -1 , but they were undetected in non-hyperaccumulator Pteris ensiformis at 10 μM AsV. Furthermore, compared to 500 μM phytate, the phytate concentration in the growth media was reduced by 69% to 155 μM, whereas the P and As contents in P. vittata fronds and roots were enhanced by 68-134% and 44-81% to 2423-2954 and 82-407 mg kg -1 under 500 μM phytate plus 50 μM AsV. The increased P/As uptake in P. vittata was probably attributed to 3.0-4.5-fold increase in expressions of P transporters PvPht1;3-1;4 . Besides, under As exposure, plant P may be converted to phytate in P. vittata roots, thereby increasing phytate's contents by 84% to 840 mg kg -1 . Overall, our results suggest that As-induced phytate/phytase exudation and phytate-P uptake stimulate its growth and As hyperaccumulation by P. vittata .

Selenate enhances arsenic (As) accumulation in As-hyperaccumulator Pteris vittata , but the associated molecular mechanisms are unclear. Here, we investigated the mechanisms of selenate-induced arsenic accumulation by exposing P. vittata to 50 μM arsenate (AsV 50 ) and 1.25 (Se 1.25 ) or 5 μM (Se 5 ) selenate in hydroponics. After 2 weeks, plant biomass, plant As and Se contents, As speciation in plant and growth media, and important genes related to As detoxification in P. vittata were determined. These genes included P transporters PvPht1;3 and PvPht1;4 (AsV uptake), arsenate reductases PvHAC1 and PvHAC2 (AsV reduction), and arsenite (AsIII) antiporters PvACR3 and PvACR3;2 (AsIII translocation) in the roots, and AsIII antiporters PvACR3;1 and PvACR3;3 (AsIII sequestration) in the fronds. The results show that Se 1.25 was more effective than Se 5 in increasing As accumulation in both P. vittata roots and fronds, which increased by 27 and 153% to 353 and 506 mg kg -1 . The As speciation analyses show that selenate increased the AsIII levels in P. vittata , with 124-282% more AsIII being translocated into the fronds. The qPCR analyses indicate that Se 1.25 upregulated the gene expression of PvHAC1 by 1.2-fold, and PvACR3 and PvACR3;2 by 1.0- to 2.5-fold in the roots, and PvACR3;1 and PvACR3;3 by 0.6- to 1.1-fold in the fronds under AsV 50 treatment. Though arsenate enhanced gene expression of P transporters PvPht1;3 and PvPht1;4 , selenate had little effect. Our results indicate that selenate effectively increased As accumulation in P. vittata , mostly by increasing reduction of AsV to AsIII in the roots, AsIII translocation from the roots to fronds, and AsIII sequestration into the vacuoles in the fronds. The results suggest that selenate may be used to enhance phytoremediation of As-contaminated soils using P. vittata .

Arsenic (As) is toxic and ubiquitous in the environment, posing a growing threat to human health. As-hyperaccumulator Pteris vittata has been used for phytoremediation of As-contaminated soil. Symbiosis with arbuscular mycorrhizal fungi (AMF) enhances As accumulation by P. vittata , which is different from As inhibition in typical plants. In this study, P. vittata seedlings inoculated with or without AMF were cultivated in As-contaminated soils for 2 months. AMF-root symbiosis enhanced plant growth, with 64.5% greater As contents in the fronds. After exposure to AsV for 2 h, the arsenate (AsV) and arsenite (AsIII) contents in AMF-roots increased by 1.8- and 3.6-fold, suggesting more efficient As uptake by P. vittata with AMF-roots. Plants take up and transport AsV via phosphate transporters (Phts). Here, for the first time, we identified a novel mycorrhiza-specific Pht transporter, PvPht1;6, from P. vittata . The transcripts of PvPht1;6 were strongly induced in AMF-roots, which were localized to the plasma membrane of arbuscule-containing cells. By complementing a yeast mutant lacking 5-Phts, we confirmed PvPht1;6's transport activity for both P and AsV. In contrast to typical AMF-inducible phosphate transporter LePT4 from tomato, PvPht1;6 showed greater AsV transport capacity. The results suggest that PvPht1;6 is probably critical for AsV transport at the periarbuscular membrane of P. vittata root cells, revealing the underlying mechanism of efficient As accumulation in P. vittata with AMF-roots.

Phytate ( myo -inositol hexakisphosphate salts) can constitute a large fraction of the organic P in soils. As a more recalcitrant form of soil organic P, up to 51 million metric tons of phytate accumulate in soils annually, corresponding to ∼65% of the P fertilizer application. However, the availability of phytate is limited due to its strong binding to soils via its highly-phosphorylated inositol structure, with sorption capacity being ∼4 times that of orthophosphate in soils. Phosphorus (P) is one of the most limiting macronutrients for agricultural productivity. Given that phosphate rock is a finite resource, coupled with the increasing difficulty in its extraction and geopolitical fragility in supply, it is anticipated that both economic and environmental costs of P fertilizer will greatly increase. Therefore, optimizing the use of soil phytate-P can potentially enhance the economic and environmental sustainability of agriculture production. To increase phytate-P availability in the rhizosphere, plants and microbes have developed strategies to improve phytate solubility and mineralization by secreting mobilizing agents including organic acids and hydrolyzing enzymes including various phytases. Though we have some understanding of phytate availability and phytase activity in soils, the limiting steps for phytate-P acquisition by plants proposed two decades ago remain elusive. Besides, the relative contribution of plant- and microbe-derived phytases, including those from mycorrhizas, in improving phytate-P utilization is poorly understood. Hence, it is important to understand the processes that influence phytate-P acquisition by plants, thereby developing effective molecular biotechnologies to enhance the dynamics of phytate in soil. However, from a practical view, phytate-P acquisition by plants competes with soil P fixation, so the ability of plants to access stable phytate must be evaluated from both a plant and soil perspective. Here, we summarize information on phytate availability in soils and phytate-P acquisition by plants. In addition, agronomic approaches and biotechnological strategies to improve soil phytate-P utilization by plants are discussed, and questions that need further investigation are raised. The information helps to better improve phytate-P utilization by plants, thereby reducing P resource inputs and pollution risks to the wider environment.

Arsenic (As) contamination in soils is of great concerns due to its toxicity to plants. As an analogue, phosphorus plays an important role in protecting plants from As toxicity. In this study, we identified a new phosphate transporter 2 (PHT2), PvPht2;1, from As-hyperaccumulator Pteris vittata and analyzed its functions in As and P transport in a yeast mutant, and model plant Arabidopsis thalian . PvPht2;1 contained 12 transmembrane domains, sharing high identity with PHT2 genes in diverse plants. Further, independent of external P or As levels, PvPht2;1 was mainly expressed in P. vittata fronds with the expression being 3-4 folds higher than that in the roots and rhizomes. Localized to the chloroplasts based on GFP-fused PvPht2;1 in model plant tobacco, PvPht2;1 functioned as a low-affinity P transporter. Under As exposure, PvPht2;1 yeast transformants showed comparable growth with the control while high-affinity P transporter PvPht1;3 transformants showed better growth, suggesting that PvPht2;1 transported P but slower than PvPht1;3 transporter. Expressing PvPht2;1 in A. thaliana increased its shoot P concentration without influencing its As accumulation. Further, the chloroplasts' P content in transgenic A. thaliana increased by 37-59% than wild-type (WT) plants. Under As exposure, the photosynthesis of PvPht2;1 -expressing A. thaliana remained stable but that of WT plants decreased. The data indicate that, under As stress, expressing PvPht2;1 in A. thaliana enhanced its P transport to the chloroplasts and protected its photosynthesis. In short, highly expressed in the fronds and not impacted by As exposure, chloroplast-located PvPht2;1 may have protected As-hyperaccumulator P. vittata from As toxicity by efficiently transporting only P to its chloroplasts.

Bioavailability and speciation of arsenic (As) are impacted by fertilization and bacteria in the rice rhizosphere. In this study, we investigated the effects of long-term manure application on As bioavailability, microbial community structure, and functional genes in a rice paddy field. The results showed that manure application did not affect total As in the soil but increased soluble As forms by 19%, increasing arsenite (As(III)) accumulation in rice grains and roots by 34 and 64% compared to a control. A real-time quantitative polymerase chain reaction (qPCR) and high-throughput sequencing analysis demonstrated that manure application increased the relative abundance of Rhizobium , Burkholderia , Sphingobium , and Sphingomonas containing arsenate reductase genes ( arsC ) in the rhizosphere soil, consistent with the 529% increase in arsC , which may have promoted arsenate (As(V)) reduction and increased As availability in pore water. In addition, manure application significantly altered the iron (Fe)-plaque microbial community structure and diversity. The microbes, particularly, Bradyrhizobium , Burkholderia , and Ralstonia , were mostly associated with As, Fe, and sulfur (S) cycles. This result was consistent with changes in the functional genes related to As, Fe, and S transformation. Although manure application promoted As(V) reduction ( arsC) in Fe-plaque by 682%, it inhibited Fe and S reduction by decreasing Fe III reduction bacteria (Geobacteraceae) and the sulfate-reducing gene ( dsrA ) abundance. Further, manure application changed the composition of the microbial community that contained the arsC gene. In short, caution needs to be excised even in the soil with a low As concentration as manure application increased As(III) accumulation in rice grains.

Incidental soil ingestion is considered to be an important route of exposure to hydrophobic organic contaminants (HOCs), such as dichlorodiphenyl-trichloroethane (DDT). Contaminant ingestion often occurs during food consumption; however, knowledge on the influence of food on DDT bioavailability remains limited. In this study, the relative bioavailability (RBA) of soil DDTr (i.e., DDT and metabolites) was determined using an in vivo mouse model in the presence of eight kinds of food including rice, egg, pork, pear, soybean, bread, spinach, and milk powder. The values of DDTr-RBA ranged from 19.8 ± 10.9 to 114 ± 25.1%. DDTr-RBA was positively correlated with fat ( r = 0.71) and negatively correlated with fiber ( r = 0.63) content in food. A mechanistic study showed that fat enhanced micellarization and promoted the formation of chylomicron, which facilitated the dissolution and transport of DDTr in the intestinal tract. Bioaccessibility of DDTr was determined using a physiologically based in vitro method. The addition of lipase significantly improved the ability of the method to predict DDTr-RBA, indicating that the "fasted state" in vitro method required optimization for food scenarios. To the best of our knowledge, this is the first study to explore the mechanistic influence of food on DDTr-RBA and provide important knowledge on dietary approaches for reducing exposure to HOCs.

Behavior of trace elements in flooded/lowland rice soils is controlled by root-zone iron oxidation. Insoluble iron species bind/capture toxic elements, i.e., arsenic. However, it was recently observed that within this territory of arsenic immobilization lies a zone of prolific iron release, accompanied by a significant flux of arsenic in close proximity to rice root apices. Questions still remain on how common this phenomenon is and whether the chemical imaging approaches or soils/cultivars used influence this event. Here, three types of ultrathin/high-resolution diffusive gradient in thin films (DGT) substrates were integrated with oxygen planar optodes in a multilayer system, providing two-dimensional mapping of solute fluxes. The three DGT approaches revealed a consistent/overlapping spatial distribution with localized flux maxima for arsenic, which occurred in all experiments, concomitant with iron mobilization. Soil/porewater microsampling within the rhizosphere revealed no significant elevation in the solid phase's total iron and arsenic concentrations between aerobic and anaerobic zones. Contrary to arsenic, phosphorus bioavailability was shown to decrease in the arsenic/iron flux maxima. Rice roots, in addition to their role in nutrient acquisition, also perform a key sensory function. Flux maxima represent a significant departure from the chemical conditions of the bulk/field environment, but our observations of a complete rhizosphere reveal a mixed mode of root-soil interactions.

Arsenic-hyperaccumulator Pteris vittata is efficient in As uptake, probably through phosphate transporters (Pht). Here, for the first time, we cloned a new PvPht1;4 gene from P. vittata and investigated its role in arsenate (AsV) uptake and transport in yeast and transgenic tobacco plants. On the basis of quantitative real-time polymerase chain reaction (qRT-PCR), PvPht1;4 was abundantly expressed in P. vittata fronds and roots, with its transcripts in the roots being induced by both P deficiency and As exposure. PvPht1;4 was localized to the plasma membrane, which complemented a yeast-mutant defective in P uptake and showed higher P transport affinity than PvPht1;3. Under AsV exposure, PvPht1;4 yeast transformants showed comparable tolerance as PvPht1;3, but higher As accumulation than PvPht1;2 transformants, indicating that PvPht1;4 had considerable AsV and P transport activity. However, in soil and hydroponic experiments, PvPht1;4 expressing tobacco lines accumulated 26-44 and 37-55% lower As in the shoots than wild type plants, with lower root-to-shoot As translocation. In the roots of PvPht1;4 lines, higher glutathione (GSH) contents and expression levels of GSH synthetase gene NtGSH2 were observed. In addition, the transcripts of AsIII-GSH transporter NtABCC1 in PvPht1;4 lines were upregulated. The data suggested that PvPht1;4 lines probably detoxified As by reducing AsV to AsIII, which was then complexed with GSH and stored in the root vacuoles, thereby reducing As translocation in transgenic tobacco. Given its strong AsV transport capacity, expression of PvPht1;4 provides a new molecular approach to reduce As accumulation in plant shoots.

Soils are often co-contaminated with As, Pb, and Cd. To what extent ingested metal(loid)s interact with each other in the gastrointestinal tract and influence their RBA (relative bioavailability) is largely unknown. Three soils predominantly contaminated with As (MS, mining/smelting impacted), Pb (WR, wire rope production impacted), and Cd (EP, enamel pottery production impacted) were administered to mice individually or in binary and tertiary combinations with sodium arsenate, Cd chloride, and/or Pb acetate. In binary combinations, ∼10-fold higher Pb addition decreased As-RBA in MS (26.0 ± 6.28% to 17.1 ± 1.08%), while ∼10-fold higher As addition decreased Pb-RBA in WR (61.3 ± 2.41% to 28.8 ± 5.45%). This was possibly due to the formation of insoluble Pb arsenate in mouse intestinal tract, as indicated by the formation of precipitates when As and Pb co-occurred in water or simulated human gastrointestinal fluids. Due to competition for shared absorption transporters, ∼10- and 100-fold higher Pb addition decreased Cd-RBA in EP (95.8 ± 12.9% to 67.8 ± 12.8% and 62.8 ± 8.24%). Tertiary combinations showed that interactions between two metal(loid)s were affected by the presence of the third metal(loid). Our study suggests that As oxyanion could interact with Pb or Cd ions in the mouse gastrointestinal tract, and the interactions vary depending on concentration and solution characteristics.

Psychiatric pharmaceuticals are widely distributed in the aquatic environment and have attracted recent attention due to their potential for environmental effects. A robust and reliable in situ passive sampling approach, the diffusive gradients in thin-films (DGT) technique, is developed here to measure 14 psychiatric pharmaceuticals. A new binding material, mixed-mode cation exchange resin (Poly-Sery MCX, 40 μm, CNW, Germany), was used for the first time in DGT and compared to XAD and HLB. Reliable elution efficiencies of the pharmaceuticals from the binding gels were obtained in methanol/ammonia, and diffusion coefficients for all the compounds were determined. The influence of diffusive layer thickness (0.515-2.015 mm), deployment time (3-168 h), and important environmental conditions-pH (3.02-9.45), ionic strength (0.0001-0.5 M), and dissolved organic matter (0-20 mg L -1 )-were evaluated. The capacity of XAD, HLB, and MCX gels for binding all the test pharmaceuticals was ∼335 μg per disc, meaning that DGT could theoretically be deployed for over 30 months if there are no competitive effects or confounding factors. The uptake kinetics of psychiatric pharmaceuticals onto MCX gel were much faster than those onto XAD and HLB gels in the first hour. DGT measured concentrations of test pharmaceuticals at two sample points in a river (over 6 days) were comparable to those obtained by grab sampling. This study demonstrates the accuracy and reliability of DGT for measuring psychiatric pharmaceuticals across a wide range of freshwater conditions found in the natural environment.

Metal contamination in soil, dust, and food matrices impacts the health of millions of people worldwide. During the past decades, various animal bioassays have been developed to determine the relative bioavailability (RBA) of As, Pb, and Cd in contaminated soils, dust, and foods, which vary in operational approaches. This review discusses the strengths and weaknesses of different animal models (swine and mice), dosing schemes (single gavage dose, repeated gavage dose, daily repeated feeding, and free access to diet), and end points (blood, urine, and tissue) in metal-RBA measurement; compares metal-RBA obtained using mouse and swine bioassays, different dosing schemes, and different end points; and summarizes key findings on As-, Pb-, and Cd-RBA values in contaminated soils, dust, and foods. Future directions related to metal-RBA research are highlighted, including (1) comparison of metal-RBA determinations between different bioassays and different laboratories to ensure robust bioavailability data, (2) enhancing the metal-RBA database for contaminated dust and foods, (3) identification of physiological and physicochemical mechanisms responsible for variability in metal-RBA values, (4) formulation of strategies to decrease metal-RBA values in contaminated soils, dust, and foods, and (5) assessing the impacts of cocontaminants on metal-RBA measurement.

Arsenic-hyperaccumulator Pteris vittata is efficient in As accumulation and has been used in phytoremediation of As-contaminated soils. Arsenate (AsV) is the predominant As species in aerobic soils and is taken up by plants via phosphate transporters (Pht) including P. vittata . In this work, we cloned the PvPht1;3 full length coding sequence from P. vittata and investigated its role in As accumulation by yeast and plants. PvPht1;3 complemented a yeast P uptake mutant strain and showed a stronger affinity and transport capacity to AsV than PvPht1;2. In transgenic tobacco, PvPht1;3 enhanced AsV absorption and translocation, increasing As accumulation in the shoots under both hydroponic and soil experiments. On the basis of the expression patterns via qRT-PCR, PvPht1;3 was strongly induced by P deficiency but not As exposure. To further understand its expression pattern, transgenic Arabidopsis thaliana and soybean expressing the GUS reporter gene, driven by PvPht1;3 promoter, were produced. The GUS staining showed that the reporter gene was mainly expressed in the stele cells, indicating that PvPht1;3 was expressed in stele cells and was likely involved in P/As translocation. Taken together, the data suggested that PvPht1;3 was a high-affinity AsV transporter and was probably responsible for efficient As translocation in P. vittata . Our results suggest that expressing PvPht1;3 enhances As translocation and accumulation in plants, thereby improving phytoremediation of As-contaminated soils.

Rice ( Oryza sativa ) is a major food crop in the world, feeding half of the world's population. However, rice is efficient in taking up toxic metalloid arsenic (As), adversely impacting human health. Among different As species, inorganic As is more toxic than organic As. Thus, it is important to decrease inorganic As in rice to reduce human exposure from the food chain. The arsenite (AsIII) antiporter gene PvACR 3;1 from As-hyperaccumulator Pteris vittata decreases shoot As accumulation when heterologously expressed in plants. In this study, three homozygous transgenic lines (L2, L4, and L7) of T3 generation were obtained after transforming PvACR 3;1 into rice. At 5 μM of AsIII, PvACR 3;1 transgenic rice accumulated 127%-205% higher As in the roots, with lower As translocation than wild type (WT) plants. In addition, at 20 μM of AsV, the transgenic rice showed similar results, indicating that expressing PvACR3;1 increased As retention in the roots from both AsIII and AsV. Furthermore, PvACR 3;1 transgenic rice plants were grown in As-contaminated soils under flooded conditions. PvACR3;1 decreased As accumulations in transgenic rice shoots by 72%-83% without impacting nutrient minerals (Mn, Zn, and Cu). In addition, not only total As in unhusked rice grain of PvACR 3;1 transgenic lines were decreased by 28%-39%, but also inorganic As was 26%-46% lower. Taken together, the results showed that expressing PvACR3;1 effectively decreased both total As and inorganic As in rice grain, which is of significance to breed low-As rice for food safety and human health.

Although strong in vivo-in vitro correlations (IVIVCs) between relative bioavailability (RBA) and bioaccessibility of soil Pb were well reported, knowledge on the fractions of bioaccessible Pb in simulated gastrointestinal (GI) fluids that are available for absorption into the systemic circulation is limited. Here, Pb-RBA in 14 Pb-contaminated soils were assessed using an in vivo mouse bioassay and compared to Pb bioaccessibility by the gastrointestinal phase of the UBM (Unified Bioaccessibility research group of Europe (BARGE) Method) in vitro assay with and without 0.45 μm filtration of GI fluid. Results showed good IVIVC between Pb-RBA and Pb bioaccessibility without filtration ( r 2 = 0.62), while Pb bioaccessibility with filtration provided a poor correlation with Pb-RBA ( r 2 = 0.16). This suggested that besides dissolved Pb ions, Pb-complexes formed in the UBM gastrointestinal fluid might also contribute to bioavailable Pb. To ascertain this, DGT (diffusive gradients in thin-films) devices which can measure both Pb 2+ ions and labile inorganic and organic Pb-complexes were introduced to the UBM fluids to measure Pb DGT-bioaccessibility, which showed strong correlation to Pb-RBA ( r 2 = 0.71). With increasing diffusive gel thickness which could enhance release of Pb ions from Pb-complexes, Pb DGT-bioaccessibility increased by 3.4-5.7 times, while inclusion of dialysis membrane within DGT devices significantly decreased Pb DGT-bioaccessibility by inhibiting diffusion of Pb complexes to binding gel. These results confirmed the contribution of Pb-complexes to Pb bioavailability, providing new insights to Pb bioavailability.

Mushrooms accumulate arsenic (As), yet As concentrations, speciation, and localization in cultivated mushrooms across a large geographic distribution are unknown. We characterized 141 samples of nine species from markets in nine capital cities in China, with samples of Lentinula edodes, Pleurotus ostreatus, and Agaricus bisporus being analyzed for As speciation and localization. Total As concentrations ranged from 0.01 to 8.31 mg kg -1 dw, with A. bisporus (0.27-2.79 mg kg -1 ) containing the most As followed by P. ostreatus and L. edodes (0.04-8.31 and 0.12-2.58 mg kg -1 ). However, As in A. bisporus was mostly organic including nontoxic arsenobetaine, while P. ostreatus and L. edodes contained mainly inorganic As (iAs). On the basis of in situ imaging using LA-ICP-MS, As in L. edodes was localized to the surface coat of the cap, while As in P. ostreatus was localized to the junction of the pileus and stipe. When As speciation and daily mushroom consumption (1.37 g d -1 dw) are considered, daily mushroom consumption may result in elevated iAs exposure, with increased bladder and lung cancer rates up to 387 cases per 100000. Our study showed that market mushrooms could be a health risk to the general public so its production should be monitored.

The speciation of selenium (Se) controls its fate and behavior, determining both its biological and environmental activities. However, in situ monitoring of Se IV presents a significant challenge due to its sensitivity to redox change. A novel diffusive gradients in thin films (DGT) technique containing mercapto-, amino-bifunctionalized SBA15 mesoporous silica nanoparticles was developed and evaluated in a series of laboratory and field deployment tests. The SBA-DGT exhibited a linear accumulation of Se IV ( r 2 > 0.997) over a 72 h deployment, with negligible accumulation of Se VI (<5%). Consistent prediction of Se IV occurred within ionic strength and pH ranges of 0.1-200 mmol L -1 and 3.6-8, respectively. Limits of detection of the SBA-DGT were 0.03 μg Se IV L -1 , which is suitable for natural waters. Moreover, the properties of the bifunctionalized SBA15 enable it to be fabricated within ultrathin (0.05 mm) gel layers for use in conjunction with O 2 planar optode imaging. This new sandwich sensor technology with SBA-DGT was validated by mapping the two-dimensional distribution of Se IV and oxygen simultaneously in rice rhizospheres. This study shows that SBA-DGT provides a selective measurement of Se IV in situ, demonstrating its potential for both environmental monitoring and as a research tool for improving our understanding of Se biogeochemical processes.

A method using miniaturized arrayed DGT-probes (PADDI) for high-frequency in situ sampling with LA-ICPMS and CID analysis was developed to measure the field-scale heterogeneity of trace-element bioavailability. Robust calibrations (R 2 > 0.99) combined with high-sensitivity (LOD = 0.35 ng cm -2 ), multielemental detection, and short measurement times were achieved using a new LA-ICPMS microDGT analysis. In the studied paddy-site (size: ∼2500 m 2 ), total element concentrations across the field were approximately uniform (R.S.D. < 10%), but bioavailability was shown to vary significantly as determined from 864 microgel measurements housed within 72 PADDI arrays. Porewater As measurements were unable to differentiate the top/rhizosphere and bulk/deeper-soil layers. However, dynamic sampling with DGT revealed significant differences. Heterogeneity behaviors varied greatly between the different elements. Arsenic bioavailability was stable laterally across the field, but varied with depth, which was in contrast to the trends for Pb. Fe/S (-II) change was bidirectional, differing horizontally and vertically throughout the field. The heterogeneity in Pb bioavailability, due to the high frequency of hotspot maxima that were discretely dispersed across the paddy, proved the most difficult to simulate requiring the greatest number of probe deployments to determine a reliable field-average. The DGT-PADDI system provides a new characterization of infield trends for improved trace-inorganics' management in agricultural wetlands.

The interactions of gaseous 2-chlorophenol with Fe 3+ -saturated montmorillonite particles in a gas-solid system were investigated to simulate the reactions of mineral dusts with volatile organic pollutants in the atmosphere. Results suggested that Fe 3+ -saturated montmorillonite mediated the dimerization of gaseous 2-chlorophenol to form hydroxylated polychlorinated biphenyl, hydroxylated polychlorinated diphenyl ether, and hydroxylated polychlorinated dibenzofuran. The toxicity of Fe 3+ -montmorillonite particles to A549 human lung epithelial cells before and after interaction with 2-chlorophenol was examined to explore their adverse impact on human health. Based on cell morphological analysis, cytotoxicity tests, and Fourier-transform infrared imaging spectra, surface-catalyzed reactions of Fe 3+ -montmorillonite with 2-chlorophenol increased the toxicity of montmorillonite particle on A549 cells. This was supported by increased cellular membrane permeability, the release of extracellular lactate dehydrogenase, and cell damages on cellular DNA, proteins, and lipids. Since mineral dusts are important components of particulate matter, our results help to understand the interactions of volatile organic pollutants with particulate matter in the atmosphere and their adverse impacts on human health.

Phosphorus is an important macronutrient for plant growth and is acquired by plants mainly as phosphate (P). Phosphate transporters (Phts) are responsible for P and arsenate (AsV) uptake in plants including arsenic-hyperaccumulator Pteris vittata. P. vittata is efficient in AsV uptake and P utilization, but the molecular mechanism of its P uptake is largely unknown. In this study, a P. vittata Pht, PvPht1;2, was cloned and transformed into tobacco ( Nicotiana tabacum). In hydroponic experiments, all transgenic lines displayed markedly higher P content and better growth than wild type, suggesting that PvPht1;2 mediated P uptake in plants. In addition, expressing PvPht1;2 also increased the shoot/root 32 P ratio by 69-92% and enhanced xylem sap P by 46-62%, indicating that PvPht1;2 also mediated P translocation in plants. Unlike many Phts permeable to AsV, PvPht1;2 showed little ability to transport AsV. In soil experiments, PvPht1;2 also significantly increased shoot biomass without elevating As accumulation in PvPht1;2 transgenic tobacco. Taken together, our results demonstrated that PvPht1;2 is a specific P transporter responsible for P acquisition and translocation in plants. We envisioned that PvPht1;2 can enhance crop P acquisition without impacting AsV uptake, thereby increasing crop production without compromising food safety.

Arsenic (As) accumulation in rice grains poses health risk to humans. Plants including rice take up arsenate (AsV) by phosphate transporters. In this study, rice phosphate transporter OsPT4 (OsPht1;4) was investigated based on two independent T-DNA insertion mutants of OsPT4 (M1 and M2), which displayed stronger AsV resistance than wild types WT1 and WT2. When cultivated in medium (+P or -P) with AsV, ospt4 mutants accumulated 16-32% lower As in plants, suggesting that OsPT4 mediates AsV uptake. Analysis of the xylem sap showed that AsV concentrations in ospt4 mutants was 20-40% lower than WT controls under -P condition, indicating OsPT4 may also mediate AsV translocation. Moreover, kinetics analysis showed that ospt4 mutants had lower AsV uptake rates than the WT controls, further proving that OsPT4 functions as an AsV transporter in rice. When grown in flooded soils with As, AsV concentrations in rice grains of ospt4 mutants decreased by 50-55%. More importantly, knocking out OsPT4 in M1 and M2 reduced inorganic As accumulation in rice grains by 20-44%, significant for controlling As exposure risk from rice. Taken together, our findings revealed a critical role of OsPT4 in AsV uptake and translocation in rice. Knocking out OsPT4 effectively decreased inorganic As accumulation in rice grains, shedding light on engineering low-As rice to enhance food safety.

To determine the effectiveness of mineral dietary supplements to modulate cadmium (Cd) exposure, an in vivo mouse bioassay was conducted to determine Cd relative bioavailability (Cd-RBA) in Cd-contaminated rice (0.80 mg Cd kg -1 ) with and without Zn, Fe, or Ca supplements as nitrate or chloride salts. Without mineral supplements, Cd-RBA was 43 ± 5.3% based on average Cd accumulation in the liver plus kidneys as the end point. Among Ca(NO 3 ) 2 , Zn(NO 3 ) 2 , and Fe(NO 3 ) 2 supplements, 150-5000 mg kg -1 Ca was the most effective in reducing rice Cd-RBA by 31-80% to 8.5-29%, while 30-200 mg kg -1 Zn supplements was ineffective, with Cd-RBA being 33-57%. Low Fe at <40 mg kg -1 had little impact on rice Cd-RBA (39-47%), while high Fe at 80-200 mg kg -1 decreased Cd-RBA by 37% to 26-27%. The ineffectiveness of Zn supplements in reducing Cd-RBA was probably due to coinciding 8.3- and 3.1-fold increases in Zn accumulation in mouse kidneys and liver with Zn supplements, while Ca and Fe supplements led to much-smaller increases in Ca and Fe accumulation in mouse tissues (1.3-1.6 fold). In addition, compared to Ca(NO 3 ) 2 supplements, Cd-RBA values determined with CaCl 2 supplements were significantly higher (25-67% versus 8.5-29%), suggesting that chloride enhanced Cd-RBA. Results of this study have important implications for developing effective dietary strategies to reduce dietary Cd exposure and the associated health risks in humans.

The stability of perfluorooctanoate (PFOA) coupled with its wide use cause serious concerns regarding its potential risk to human health. The molecular mechanisms of PFOA-induced hepatotoxicity relevant to human health was investigated using both in vivo (mouse model) and in vitro (human hepatocyte cells, HL-7702) techniques. Both male and female Balb/c mice were administered PFOA at 0.05, 0.5, or 2.5 mg/kg-d for 28-d, with serum PFOA concentrations after exposure being found at environmentally relevant levels. Liver samples were examined for histology and proteomic change using iTRAQ and Western Blotting, showing dose-dependent hepatocyte apoptosis and peroxisome proliferation. At high doses, genotoxicity resulting from ROS hypergeneration was due to suppression of Complex I subunits in the electron transport chain and activation of PPARα in both genders. However, at 0.05 mg/kg-d, Complex I suppression occurred only in females, making them more sensitive to PFOA-induced apoptosis. In vitro assays using HL-7702 cells confirmed that apoptosis was also induced through a similar mechanism. The dose/gender-dependent toxicity mechanisms help to explain some epidemiological phenomena, i.e., liver cancer is not often associated with PFOA exposure in professional workers. Our results demonstrated that a proteomic approach is a robust tool to explore molecular mechanisms of toxic chemicals at environmentally relevant levels.

Arsenic (As) is a toxic carcinogen so it is crucial to decrease As accumulation in crops to reduce its risk to human health. Arsenite (AsIII) antiporter ACR3 protein is critical for As metabolism in organisms, but it is lost in flowering plants. Here, a novel ACR3 gene from As-hyperaccumulator Pteris vittata, PvACR3;1, was cloned and expressed in Saccharomyces cerevisiae (yeast), Arabidopsis thaliana (model plant), and Nicotiana tabacum (tobacco). Yeast experiments showed that PvACR3;1 functioned as an AsIII-antiporter to mediate AsIII efflux to an external medium. At 5 μM AsIII, PvACR3;1 transgenic Arabidopsis accumulated 14-29% higher As in the roots and 55-61% lower As in the shoots compared to WT control, showing lower As translocation. Besides, transgenic tobacco under 5 μM AsIII or AsV also showed similar results, indicating that expressing PvACR3;1 gene increased As retention in plant roots. Moreover, observation of PvACR3;1-green fluorescent protein fusions in transgenic Arabidopsis showed that PvACR3;1 protein localized to the vacuolar membrane, indicating that PvACR3;1 mediated AsIII sequestration into vacuoles, consistent with increased root As. In addition, soil experiments showed ∼22% lower As in the shoots of transgenic tobacco than control. Thus, our study provides a potential strategy to limit As accumulation in plant shoots, representing the first report to decrease As translocation by sequestrating AsIII into vacuoles, shedding light on engineering low-As crops to improve food safety.

Dietary Cd intake is often estimated without considering Cd bioavailability. Measured urinary Cd for a cohort of 119 nonsmokers with rice as a staple was compared to predicted values from rice-Cd intake with and without considering Cd relative bioavailability (RBA) in rice based on a steady state mouse kidney bioassay and toxicokinetic model. The geometric mean (GM) of urinary Cd and β 2 -microglobulin was 1.08 and 234 μg g -1 creatinine. Applying Cd-RBA in foods to aggregate Cd intake (41.5 ± 12.4, 48.0 ± 9.3, 48.8 ± 21.3% for rice, wheat, and vegetables), rice was the largest contributor (71%). For 63 participants providing paired urine and rice samples, the predicted GM of urinary Cd at 4.14 μg g -1 based on total Cd in rice was 3.5 times that of measured value at 1.20 μg g -1 , while incorporating Cd-RBA to assess rice-Cd intake made the two closer with GM at 1.07 μg g -1 . The cohort findings were extended to a national scale, with urinary Cd for nonsmokers from rice Cd intake was mapped at province/city levels after considering rice Cd-RBA. Therefore, incorporating Cd bioavailability to assess dietary Cd intake is a valuable tool to accurately estimate human Cd exposure and associated health risk.

A steady-state mouse model was developed to determine arsenic (As) relative bioavailability (RBA) in rice to refine As exposure in humans. Fifty-five rice samples from 15 provinces of China were analyzed for total As, with 11 cooked for As speciation and bioavailability assessment. Arsenic concentrations were 38-335 μg kg -1 , averaging 133 μg kg -1 , with As III being dominant (36-79%), followed by DMA V (18-58%) and As V (0.5-16%). Following oral doses of individual As species to mice at low As exposure (2.5-15 μg As per mouse) over a 7-d period, strong linear correlations (R 2 = 0.99) were observed between As urinary excretion and cumulative As intake, suggesting the suitability and sensitivity of the mouse bioassay to measure As-RBA in rice. Urinary excretion factor for DMA V (0.46) was less than inorganic As (0.63-0.69). As-RBA in cooked rice ranged from 13.2 ± 2.2% to 53.6 ± 11.1% (averaging 27.0 ± 12.2%) for DMA V and 26.2 ± 7.0% to 49.5 ± 4.7% (averaging 39.9 ± 8.3%) for inorganic As. Calculation of inorganic As intake based on total inorganic As in rice overestimated As exposure by 2.0-3.7 fold compared to that based on bioavailable inorganic As. For accurate assessment of the health risk associated with rice consumption, it is important to consider As bioavailability especially inorganic As in rice.

Arsenic hyperaccumulator Pteris vittata (PV) is efficient in taking up As and nutrients from As-contaminated soils. We evaluated the mechanisms used by PV to mobilize As and Fe by examining the impacts of As and root exudates on FeAsO4 solubilization, and As and Fe uptake in four plants: As-hyperaccumulators PV and Pteris multifida (PM), nonhyperaccumulator Pteris ensiformis (PE), and angiosperm plant tomato (Solanum lycopersicum). Phytate and oxalate were dominant in fern plants (>93%), which were 50-83, 15-42, and 0-32 mg kg(-1) phytate and 10-15, 7-26, and 4-12 mg kg(-1) oxalate for PV, PM, and PE respectively, with higher As inducing greater phytate exudation and no phytate being detected in tomato exudates. PV treated with phytate+FeAsO4 had higher As and Fe contents and larger biomass than phytate or FeAsO4 treatment, which were 340 vs 20 and 130 mg kg(-1) As in the fronds and 7900 vs 1600 and 4100 mg kg(-1) Fe in the roots. We hypothesized that As-induced phytate exudation helped PV to take up Fe and As from insoluble FeAsO4 and promoted PV growth. Our study suggests that phytate exudation may be special to fern plants, which may play an important role in enhancing As and nutrient uptake by plants, thereby increasing their efficiency in phytoremediation of As-contaminated soils.

Recent studies have investigated lead (Pb) concentrations in lip products but little is known about its oral bioavailability. In this study, 75 lipsticks and 18 lip glosses were assessed for Pb concentration, while 15 samples were assessed for Pb relative bioavailability (RBA, relative to Pb acetate absorption) using a mouse femur assay. Lead concentrations were 0.2-10 185 mg kg(-1), with 21 samples exceeding the Chinese limit of 40 mg kg(-1). Samples with orange and pink colors and/or low cost contained higher Pb concentrations. For samples with Pb > 7500 mg kg(-1), Pb was present due to the addition of lead chromate (PbCrO4) as a colorant, which was confirmed by X-ray absorption near-edge structure analysis. Lead-RBA in 15 samples (87-10 185 mg kg(-1)) ranged from 23% to 95%, being significantly higher in moderate Pb (56-95%; 87-300 mg kg(-1)) than high Pb samples (23-48%; >300 mg kg(-1)). The calculation of Pb intake based on Pb-RBA showed that lip product ingestion contributed 5.4-68% of the aggregate Pb exposure for women depending on Pb concentration. The high Pb concentration in some lip products together with their moderate Pb-RBA suggests that lip product ingestion is a potential health concern to women.

Using in vitro bioaccessibility assays to predict Pb relative bioavailability (RBA) in contaminated soils has been demonstrated, however, limited research was performed on urban soils having lower Pb levels. In this study, 162 soils from urban parks in 27 capital cities in China were measured for Pb bioaccessibility using the SBRC assay, with Pb-RBA in 38 subsamples being measured using a mouse-kidney assay. Total Pb concentrations in soils were 9.3-1198 mg kg(-1), with 92% of the soils having Pb concentrations <100 mg kg(-1). Lead bioaccessibility in soils was 20-94%, increasing with Pb concentration up to 100 mg kg(-1) (r = 0.44), however, limited variability in Pb bioaccessibility (60-80%) was observed for soils with Pb > 100 mg kg(-1). On the basis of a stable isotope fingerprinting technique, coal combustion ash was identified as the major Pb source, contributing to the increased Pb bioaccessibility with increasing soil Pb concentration. Lead-RBA in soils was 17-87%, showing a strong linear correlation with Pb bioaccessibility (r(2) = 0.61), with cross validation of the correlation based on random subsampling and leave-one-out approaches yielding low prediction errors. On the basis of the large sample size of 38 soils, this study demonstrated that the Pb-RBA predictive capability of the SBRC assay can be extended from mining/smelting impacted soils to urban soils with lower Pb levels.

Due to their static nature, physiologically based in vitro assays often fail to provide sufficient sorption capacity for hydrophobic organic contaminants (HOCs). The addition of a sorption sink to in vitro intestinal solution has the potential to mimic dynamic intestinal uptake for HOCs, thereby increasing their desorption from soil. However, the effectiveness of sorption sinks for improving in vitro assays needs to be compared with in vivo data. In this study, Tenax was added as a sorption sink into the physiologically based extraction test (PBET), while DDT and its metabolites (DDTr) were investigated as typical HOCs. Tenax added at 0.01-0.2 g to the PBET intestinal solution sorbed ∼100% of DDTr in 6.3-19 min, indicating its ability as an effective sorption sink. DDTr bioaccessibility in six contaminated soils using Tenax-improved PBET (TI-PBET; 27-56%) was 3.4-22 fold greater than results using the PBET (1.2-15%). In vivo DDTr relative bioavailability (RBA) was measured using a mouse adipose model with values of 17.9-65.4%. The inclusion of Tenax into PBET improved the in vivo-in vitro correlation from r(2) = 0.36 (slope = 2.1 for PBET) to r(2) = 0.62 (slope = 1.2 for TI-PBET), illustrating that the inclusion of Tenax as a sorption sink improved the in vitro prediction of DDTr RBA in contaminated soils.

Different animals and biomarkers have been used to measure the relative bioavailability of arsenic (As-RBA) in contaminated soils. However, there is a lack of As-RBA comparison based on different animals (i.e., swine and mouse) and biomarkers [area under blood As concentration curve (AUC) after a single gavaged dose vs steady-state As urinary excretion (SSUE) and As accumulation in liver or kidney after multiple doses via diet]. In this study, As-RBA in 12 As-contaminated soils with known As-RBA via swine blood AUC model were measured by mouse blood AUC, SSUE, and liver and kidney analyses. As-RBA ranges for the four mouse assays were 2.8-61%, 3.6-64%, 3.9-74%, and 3.4-61%. Compared to swine blood AUC assay (7.0-81%), though well correlated (R(2) = 0.83), the mouse blood AUC assay yielded lower values (2.8-61%). Similarly, strong correlations of As-RBA were observed between mouse blood AUC and mouse SSUE (R(2) = 0.86) and between urine, liver, and kidney (R(2) = 0.75-0.89), suggesting As-RBA was congruent among different animals and end points. Different animals and biomarkers had little impact on the outcome of in vivo assays to validate in vitro assays. On the basis of its simplicity, mouse liver or kidney assay following repeated doses of soil-amended diet is recommended for future As-RBA studies.

Hexavalent chromium (Cr(VI)) is much more toxic and mobile than the trivalent species (Cr(III)) and consequently, in situ monitoring of Cr(VI) can improve the understanding of Cr biogeochemistry and toxicity in ecosystems. The passive diffusive gradients in thin-films (DGT) technique is a powerful tool for determining metal(loid) speciation, but a binding phase that absorbs only one specific species of Cr is needed. N-Methyl-d-glucamine (NMDG) functional resin was incorporated into the DGT binding phase for selective measurement of Cr(VI). This NMDG-DGT sampler exhibited a theoretically linear accumulation of Cr(VI), with negligible accumulation (<5%) of Cr(III), even after 72 h deployment. The good prediction of Cr(VI) concentration in synthetic freshwater with NMDG-DGT, even in the presence of 10-time more Cr(III), further indicated the sampler's reliability in selective detection of Cr(VI). Moreover, its high capacity for Cr(VI), which exceeded 230 μg cm(-2), facilitates measurement of Cr(VI) in both uncontaminated natural waters and in slightly and heavily contaminated (ppm level) waters. Field deployment of the NMDG-DGT sampler in such waters allowed accurate measurement of time-averaged Cr(VI) concentration, indicating its robustness for in situ measurements of Cr speciation and its potential for further application in the risk assessment of Cr.

Fingerprinting based on stable isotopes of lead (Pb) in blood and environmental media helps to identify Pb exposure pathways in children. However, previous studies used stable isotopes of total Pb in media. In this study, a wire rope production town in China (Zhuhang) was selected for investigating the effectiveness of using isotope ratios in bioaccessible Pb to identify childhood Pb exposure pathways. Blood Pb levels of 115 children in Zhuhang were 1.7-20.4 μg dL(-1), averaging 6.1 ± 3.2 μg dL(-1) (mean ± standard deviation), and were ∼1.6 times the national average in China (3.9 ± 1.8 μg dL(-1)). Among different environmental media (housedust, soil, PM10, vegetables, rice, and drinking water), housedust (695 ± 495 mg kg(-1)) and vegetables [0.36 ± 0.40 mg (kg of fresh weight)(-1)] contained elevated Pb concentrations. The isotope ratios ((207)Pb/(206)Pb and (208)Pb/(206)Pb) of total Pb were the highest in housedust (0.8587 ± 0.0039 and 2.1049 ± 0.0087) but lower than blood Pb ratios (0.8634 ± 0.0027 and 2.1244 ± 0.0061). When using bioaccessible Pb in housedust (0.8639 ± 0.0018 and 2.1171 ± 0.0036), the isotope ratios overlapped with blood Pb ratios, suggesting that incidental ingestion of housedust was the predominant contributor to children's blood Pb. Coupling the stable isotope technique with bioaccessible Pb is more reliable for identifying Pb exposure pathways than total Pb determinations.

Water-sediment exchange is a fundamental component of oxyanion cycling in the environment. Yet, many of the (im)mobilization processes overlay complex spatial and temporal redox regimes that occur within millimeters of the interface. Only a few methods exist that can reliably capture these porewater fluxes, with the most popular being high-resolution diffusive gradients in thin films (HR-DGT). However, functionality of HR-DGT is restricted by the availability of suitable analyte binding agents within the sampler, which must be simple to cast and homogeneously distributed in the binding layer, exhibit adequate sorption capacities, be resistive to chemical change, and possess a very fine particle size (≤10 μm). A novel binding layer was synthesized to meet these requirements by in situ precipitation of zirconia into a precast hydrogel. The particle diameter≤0.2 μm of zirconia in this precipitated gel was uniform and at least 50-times smaller than the conventional molding approach. Further, this gel had superior binding and stability characteristics compared with the commonly used ferrihydrite HR-DGT technique and could be easily fabricated as an ultrathin gel (60 μm) for simultaneous oxygen imaging in conjunction with planar-optodes. Chemical imaging of anion and oxygen fluxes using the new sampler were evaluated on Lake Taihu sediments.

Food is a major source of human exposure to perfluorooctanoic acid (PFOA), however, PFOA bioavailability in food has not been studied. An in vivo mouse model and three in vitro methods (unified BARGE method, UBM; physiologically based extraction test, PBET; and in vitro digestion method, IVD) were used to determine the relative bioavailability and bioaccessibility of PFOA in the presence of 17 foods. PFOA was mixed with foods of different nutritional compositions and fed to mice over a 7-d period. PFOA relative bioavailability was determined by comparing PFOA accumulation in the liver following PFOA exposure via food to that in water. PFOA bioavailability relative to water ranged from 4.30 ± 0.80 to 69.0 ± 11.9% and was negatively correlated with lipid content (r = 0.76). This was possibly due to competitive sorption of free fatty acids with PFOA onto transporters on intestine epithelial cells. Besides, cations in the gastrointestinal tract, such as Ca(2+) and Mg(2+), are capable of complexing PFOA and partitioning to the lipid phase. On the other hand, when assessed using in vitro assays, PFOA bioaccessibility varied with methods, being 8.7-73% (UBM), 9.8-99% (PBET), and 21-114% (IVD). PFOA bioaccessibility was negatively correlated with lipid content when assessed using UBM (r = 0.82); however, a poor correlation with food composition was observed for PBET and IVD (r = 0.01-0.50). When in vivo and in vitro data were compared, a strong correlation was observed for UBM (r = 0.79), but poor relationships were observed for PBET and IVD (r = 0.11-0.22). This was probably because the higher lipolysis ability and presence of Ca(2+) and Mg(2+) in the gastrointestinal fluid of UBM resulted in a lower potential to form stable micelles compared to PBET and IVD. These results indicated that PFOA relative bioavailability was mainly affected by lipid content in foods, and UBM has the potential to determine PFOA bioaccessibility in food samples.

Incidental ingestion of household dust is an important arsenic (As) exposure pathway for children. However, compared to soils, assessment of As relative bioavailability (RBA) in household dust is limited. In this study, As-RBA in 12 household dust samples (7–38 mg kg(–1)) was measured using an in vivo mouse model and compared to As bioaccessibility determined using 4 assays [Solubility Bioaccessibility Research Consortium method (SBRC), in vitro gastrointestinal method (IVG), Deutsches Institut für Normunge.V. method (DIN), and physiologically based extraction test (PBET)]. Arsenic RBA ranged from 21.8 ± 1.6 to 85.6 ± 7.2% with samples containing low Fe and high total organic carbon content having higher As-RBA. Strong in vivo–in vitro correlations (IVIVC) were found between As-RBA and As bioaccessibility for SBRC and DIN (r2 = 0.63–0.85), but weaker ones were obtained for IVG and PBET (r2 = 0.29–0.55). The developed IVIVC for SBRC and DIN were used to calculate As-RBA based on As bioaccessibility for an additional 12 household dust samples. Although As bioaccessibility differed significantly (up to 7.7-fold) based on in vitro methods, predicted As-RBA was less variable (up to 3.0-fold) when calculated using As bioaccessibility data and the corresponding IVIVC. Our data suggested that both SBRC and DIN had potential to assess As bioavailability in household dust samples; however, additional research is needed.

House dust samples containing 25-738 mg of Pb kg(-1) from 15 cities in China were assessed for in vitro Pb bioaccessibility and in vivo Pb relative bioavailability. On the basis of stable Pb isotope ratios, the Pb in dust samples mainly originated from coal combustion. Lead bioaccessibility was determined using gastric (GP) and intestinal phase (IP) of solubility bioaccessibility research consortium (SBRC), in vitro gastrointestinal (IVG), Deutsches Institut für Normunge.V. (DIN), and physiologically based extraction test methods (PBET), while Pb relative bioavailability (RBA) was determined using a mouse blood model. Lead bioaccessibility in 24 house dust samples varied significantly (23-99%) depending on the methods. Values from the IP were considerably lower than those from the GP because of the co-precipitation of Pb with iron and re-adsorption onto the dust matrix. The SBRC assay with lower GP pH produced higher Pb bioaccessibility because of enhanced Pb dissolution. When compared to mouse blood data using 12 dust samples (29-60%), SBRC-GP and DIN-GP data were correlated with Pb RBA with r(2) values of 0.68 and 0.85 and intercepts 3.15 and 17.4, respectively. Overall, SBRC-GP had potential to predict Pb RBA in dust samples. However, our data suggested that more research is needed to develop a valid in vitro method for predicting Pb RBA in house dust.

We evaluated the ability of As-hyperaccumulator Pteris vittata (PV) to remove As from As-contaminated soils over five harvests in 2.5 years in raised beds (162 kg soil/bed). We tested the hypothesis that a P-limiting environment would enhance PV growth and As uptake owing its unique ability to uptake P under As-rich environment. In Dec. 2009, PV was transplanted to three As-contaminated soils (pH of 5.5-7.2) containing 25-129 mg kg(-1) As, which was amended with sparingly-soluble phosphate rock (PR-soil) or soluble P fertilizer (P-soil). During the 2.5-year, PV obtained sufficient P (1882 vs 2225 mg kg(-1)) from PR-soils, with increased root biomass (33%) and root exudation (53%) compared to P-soils. In addition, its frond biomass increased by 20% consecutively with each harvest (six month interval) from 18 to 36 g plant(-1). Its frond biomass in PR-soils (52.2 g plant(-1) year(-1) or ∼12 mt ha(-1) year(-1)) averaged 39% more than that in P-soils. To our knowledge, this represented the largest PV frond biomass reported, demonstrating the unique ability of PV in using insoluble P from PR in alkaline soils. In addition to biomass increase, PV from PR-soils had ∼1.5 times more As in fronds (2540, 780, and 920 mg kg(-1)) than those from P-soils (1740, 570, and 400 mg kg(-1)), with soils containing 129, 25, and 30 mg kg(-1) As, respectively. The low available P in PR-soils induced substantial plant growth and As uptake by PV. This translated into significantly more As removal from soil, averaging 48% reduction in PR-soils and 36% in P-soils in 2.5 years. With multiple harvests and PR amendments, our results showed As removal by PV from contaminated soils was ∼7 times faster than published studies.

Arsenate interferes with enzymatic processes and inhibits inorganic phosphorus (Pi) uptake in many plants. This study examined the role of phytase and phosphatase in arsenate tolerance and phosphorus (P) acquisition in the arsenic hyperaccumulator Pteris vittata . Enzyme-mediated hydrolysis of phytate in P. vittata extracts was not inhibited by arsenate at 5 mM or by heating at 100 °C for 10 min. Root exudates of P. vittata exhibited the highest phytase activity (18 nmol Pi mg(-1) protein min(-1)) when available P was low, allowing its growth on media amended with phytate as the sole source of P. Phosphorus concentration in P. vittata gametophyte tissue grown on phytate was equivalent to plants grown with inorganic phosphate at 2208 mg kg(-1), and arsenic was increased from 1777 to 2630 mg kg(-1). After 2 h of mixing with three soils, P. vittata phytase retained more activity, decreasing from ∼ 26 to ∼ 25 nmol Pi mg(-1) protein min(-1), whereas those from Pteris ensiformis and wheat decreased from ∼ 18 to ∼ 1 nmol Pi mg(-1) protein min(-1). These results suggest P. vittata has a uniquely stable phytase enabling its P acquisition in P-limiting soil environments. Furthermore, the P. vittata phytase has potential use as a soil amendment, a transgenic tool, or as a feed additive supplement, reducing the need for nonrenewable, polluting P fertilizers.

We investigated the mechanisms of Hg sorption onto biochars produced from Brazilian pepper (BP; Schinus terebinthifolius) at 300, 450, and 600 °C using different analytical techniques. The Hg sorption capacity of BP300, BP450, and BP600 was 24.2, 18.8, and 15.1 mg g(-1) based on Langmuir isotherm. FTIR data suggested the participation of phenolic hydroxyl and carboxylic groups in Hg sorption by biochars. XPS analysis showed that 23-31% and 77-69% of sorbed Hg was associated with carboxylic and phenolic hydroxyl groups in biochars BP300-450, whereas 91% of sorbed Hg was associated with a graphite-like domain on an aromatic structure in BP600 biochar, which were consistent with flow calorimetry data. Based on flow calorimetry, sorption of K and Ca onto biochar was exchangeable with the molar heat of sorption of 3.1 kJ mol(-1). By comparison, Hg sorption was via complexation with functional groups as it was not exchangeable by K or Ca with molar heat of sorption of -19.7, -18.3, and -25.4 kJ mol(-1) for BP300, BP450, and BP600. Our research suggested that Hg was irreversibly sorbed via complexation with phenolic hydroxyl and carboxylic groups in low temperature biochars (BP300 and BP450) and graphite-like structure in high temperature biochar (BP600).

Microbes play an important role in arsenic transformation and cycling in the environment. Microbial arsenic oxidation and reduction were demonstrated in the growth media of arsenic hyperaccumulator Pteris vittata L. All arsenite (AsIII) at 0.1 mM in the media was oxidized after 48 h incubation. Oxidation was largely inhibited by antibiotics, indicating that bacteria played a dominant role. To identify AsIII oxidizing bacteria, degenerate primers were used to amplify ∼500 bp of the AsIII oxidase gene aioA (aroA) using DNA extracted from the media. One aioA (aroA)-like sequence (MG-1, tentatively identified as Acinetobacter sp.) was amplified, exhibiting 82% and 91% identity in terms of gene and deduced protein sequence to those from Acinetobacter sp. 33. In addition, four bacterial strains with different arsenic tolerance were isolated and identified as Comamonas sp.C-1, Flavobacterium sp. C-2, Staphylococcus sp. C-3, and Pseudomonas sp. C-4 using carbon utilization, fatty acid profiles, and/or sequencing 16s rRNA gene. These isolates exhibited dual capacity for both AsV reduction and AsIII oxidation under ambient conditions. Arsenic-resistant bacteria with strong AsIII oxidizing ability may have potential to improve bioremediation of AsIII-contaminated water using P. vittata and/or other biochemical strategies.