Effect of external iron and arsenic species on chelant-enhanced iron bioavailability and arsenic uptake in rice (Oryza sativa L.)

This study was conducted to investigate the effect of external iron status and arsenic species on chelant-enhanced iron bioavailability and arsenic uptake. Rice seedlings (Oryza sativa L.) were used as model plant, and were grown in artificially contaminated sandy soils irrigated with Murashige and Skoog (MS) culture solution. Arsenate uptake in roots and shoots of rice seedlings were affected significantly (p > 0.05) while dimethylarsinic acid (DMAA) was not by the additional iron and chelating ligand treatments. Regardless of iron concentrations in the soil solution, HIDS increased arsenic uptake for roots more than EDTA and EDDS. Chelating ligands and arsenic species also influenced iron uptake in rice roots. Irrespective of arsenic species, HIDS was found to be more effective in the increase of iron bioavailability and uptake in rice roots compared to other chelants. There was a significant positive correlation (r = 0.78, p < 0.05) between arsenate and iron concentrations in the roots of rice seedlings grown with or without additional iron indicating that arsenate inhibit iron uptake. In contrast, there was no correlation between iron and DMAA uptake in roots. Poor correlation between iron and arsenic in shoots indicated that iron uptake in shoots was neither affected by additional iron nor by arsenic species. Compared to the control, chelating ligands increased iron uptake in shoots of rice seedlings significantly (p < 0.05). Regardless of additional iron and arsenic species, iron uptake in rice shoots did not differed among EDTA, EDDS, and HIDS treatments.     

Arsenite transporters expression in rice (Oryza sativa L.) associated with arbuscular mycorrhizal fungi (AMF) colonization under different levels of arsenite stress

As a silicon hyperaccumulator, lowland rice takes up higher levels of As than many other plants due to silicic acid and arsenite sharing the same transporters (Lsi1 and Lsi2). Glomus intraradices (AH01) was inoculated to rice under different arsenite concentrations (0, 2 and 8 μM) in order to investigate the interactions between arbuscular mycorrhizal fungus and rice on the accumulation of arsenite. The relative mRNA expressions of Lsi1 and Lsi2 resulted in a down-regulating trend in mycorrhizal plants. Under 2 μM arsenite treatments, Lsi1 and Lsi2 were significantly decreased, by 0.7-fold (P < 0.05) and 0.5-fold (P < 0.01), respectively, in mycorrhizal plants when compared with non-mycorrhizal plants. This led to the decrease of arsenite uptake per unit of root dry mass. No organic As species were detected in both roots and shoots. The As(III)/As(V) ratios indicated that mycorrhizal plants immobilized most of the arsenite proportion in the roots and prevented its translocation from the roots to the shoots.     

Salicylic acid involved in the regulation of nutrient elements uptake and oxidative stress in Vallisneria natans (Lour.) Hara under Pb stress

In this study, the alterations in nutrient elements content, reactive oxygen species level and antioxidant response were studied in leaves of Vallisneria natans (Lour.) Hara exposed to salicylic acid (SA, 10 or 100 μM), or Pb (50 μM) or their combinations for 4 d. No significant alterations in Mn and Ca content were observed but content of Cu, Zn, Fe and P decreased in plants exposed to SA alone. SA application inhibited the uptake of Pb and partially reversed Pb-induced the alterations in Mn, Ca and Fe content in leaves of V. natans exposed to 50 μM Pb. The decreased chlorophyll (a + b) and increased malondialdehyde and O²⁻ and H₂O₂ content were detected in plants exposed to 100 μM SA, 50 μM Pb, 10 μM SA + 50 μM Pb or 100 μM SA + 50 μM Pb. Application SA partially inhibited Pb-induced the increase of malondialdehyde, O²⁻ and H₂O₂ content. 100 μM SA decreased the activity of NADH oxidase and the content of non-protein thiols, carotenoids and ascorbic acid and increased the content of dehydroascorbate in plants treated with or without Pb. SA alone decreased the ascorbate peroxidase activity and increased the catalase and peroxidase activity, while SA application increased catalase activity but had no significant effect on ascorbate peroxidase and peroxidase activity in V. natans exposed to Pb. The results indicate that SA involves in the regulation of Pb uptake, nutrient balance and oxidative stress.     

Arsenic accumulation and translocation in the submerged macrophyte Hydrilla verticillata (L.f.) Royle

Worldwide contamination of arsenic in aquatic systems requires the development of a cost-effective, in situ phytoremediation technology. Hydrilla verticillata (L.f.) Royle, a submerged macrophyte widely distributed throughout the world, has the potential to effectively remove heavy metals from water. In order to understand the potential of H. verticillata for As phytofiltration and its impacts on As cycling in the water system, we investigated As accumulation, speciation and translocation in H. verticillata plants. Plant shoots showed a significant accumulation of As, with a maximum of >700 μg g⁻¹ dry weight (DW) after exposure to 20 μM arsenate [As(V)] or arsenite [As(III)] for 4 d, with no significant differences between the As(V) and As(III) treatments (P > 0.05). In addition, results of an in planta transport experiment showed that, after exposure of root and shoot to 2 μM As(V) and As(III) for 4 d, the bioconcentration factor (BCF) in roots for As(V) was almost twofold than that of As(III). Higher As BCFs in roots compared to shoots was also observed. Arsenic accumulated primarily in the cell walls of root cells (>73% of the total As in roots) and in the soluble parts of leaves (>60% of the total As in leaves). Regardless of the form of As supplied [As(III) or As(V)], As(V) was the dominant form in roots and As(III) was the dominant form in leaves. Further, basipetal translocation of As in this plant (?17%) was markedly higher than acropetal translocation (?3%). Because of accumulation of As in the shoot and immobilization of As below ground in roots, H. verticillata is a potential As phytofiltrator for bioremediation.     

Enhanced phytoextraction of Pb and other metals from artificially contaminated soils through the combined application of EDTA and EDDS

Chemically enhanced phytoextraction is achieved by the application of chelates to soils. Using pot experiments, the effect of the combined application of EDTA and EDDS on the uptake of Cu, Pb, Zn and Cd by Zea mays L. was studied. Among the tested application ratios of 1:1, 1:2, and 2:1 (EDTA/EDDS), 2:1 of EDTA:EDDS was the most efficient ratio for increasing the concentrations of Cu, Pb, Zn and Cd in the shoots. The combined application of 3.33 mmol kg(-1) soil of EDTA+1.67 mmol kg(-1) soil of EDDS produced 650 mg kg(-1) of Pb in the shoots, which was 2.4 and 5.9 times the concentration of Pb in the shoots treated with 5 mmol kg(-1) of EDTA and EDDS alone, respectively. The total phytoextraction of Pb reached 1710 microg kg(-1) soil, which was 2.1 and 6.1 times the total Pb from 5 mmol kg(-1) EDTA and EDDS alone, respectively. The combined application of EDTA and EDDS also significantly increased the translocation of Pb from the roots to the shoots. The mechanism of enhancing the phytoextraction of Pb by the combined application of EDTA+EDDS did not involve a change in the pH of the soil. The increase in the phytoextraction of Pb by the shoots of Z. mays L. was more pronounced than the increase of Pb in the soil solution with the combined application of EDTA and EDDS. It was thought that the major role of EDDS might be to increase the uptake and translocation of Pb from the roots to the shoots of plants.     

Can time-weighted average concentrations be used to assess the risks of metsulfuron-methyl to Myriophyllum spicatum under different time-variable exposure regimes?

We tested the effects of the herbicide metsulfuron-methyl on growth of the submerged macrophyte Myriophyllum spicatum under laboratory conditions using different exposure scenarios. The exposures of each scenario were comparable in the concentration × time factor, viz., the same 21-d time-weighted average (TWA) concentrations but variable in peak exposure concentrations (ranging from 0.1 to 21 000 ng ai L⁻¹) and exposure periods (1, 3, 7, 14 or 21 d). To study recovery potential of the exposed M. spicatum plants we continued the observation on shoot and root growth for another 21 d in herbicide-free medium so that the total observation period was 42 d. Non-destructive endpoints, length and number of new shoots and roots, were determined weekly from day 14 onwards. Destructive endpoints, dry-weight (DW) of main shoots, new shoots and new roots, were measured at the end of the experiment (t = 42 d).Metsulfuron-methyl exposure in particular inhibited new tissue formation but was not lethal to main shoots. On days 21 and 42 after start exposure, EC₁₀/EC₅₀ values for new tissues expressed in terms of peak concentration (=measured concentration during exposure periods of different length) showed large differences between exposure scenarios in contrast to EC₁₀/EC₅₀ values for days 21 and 42 expressed in terms of 21-d and 42-d TWA concentrations, respectively. At the end of the experiment (day 42), 42-d TWA EC_x values were remarkably similar between exposure scenarios, while a similar trend could already be observed on day 21 for 21-d TWA EC_x values. For the macrophyte M. spicatum and exposure to the herbicide metsulfuron-methyl the TWA approach seems to be appropriate to use in the risk assessment. However, the data from the toxicity experiment suggest that on day 21 also the absolute height of the pulse exposure played a (minor) role in the exposure - response relationships observed.     

Investigation of arsenic accumulation and tolerance potential of Sesuvium portulacastrum (L.) L

Sesuvium portulacastrum (L.) L., a facultative halophyte, is considered a suitable candidate for the phytoremediation of metals. An investigation of As accumulation and tolerance was conducted in Sesuvium plants upon exposure to As(V) (100-1000 μM) for 30 d. Plants demonstrated a good growth even after prolonged exposure (30 d) to high As(V) concentrations (1000 μM) and a significant As accumulation (155 μg g⁻¹ dry weight) with a bioaccumulation factor of more than ten at each concentration. The results of shoot and root dry weight, malondialdehyde accumulation, photosynthetic pigments, and total soluble proteins demonstrated that plants did not experience significant toxicity even at 1000 μM As(V) after 30 d. However, metabolites (total non-protein thiols and cysteine) and enzymes (serine acetyltransferase, cysteine synthase and γ-glutamylcysteine synthetase) of thiol metabolism, in general, remained either unaffected or showed slight decline. Hence, plants tolerated high As(V) concentrations without an involvement of thiol metabolism as a major component. Taken together, the results indicate that plants are potential As accumulator and may find application in the re-vegetation of As contaminated sites.     

Immunotoxicity in ascidians: antifouling compounds alternative to organotins: III--the case of copper(I) and Irgarol 1051

After the widespread ban of TBT, due to its severe impact on coastal biocoenoses, mainly related to its immunosuppressive effects on both invertebrates and vertebrates, alternative biocides such as Cu(I) salts and the triazine Irgarol 1051, the latter previously used in agriculture as a herbicide, have been massively introduced in combined formulations for antifouling paints against a wide spectrum of fouling organisms. Using short-term (60 min) haemocyte cultures of the colonial ascidian Botryllus schlosseri exposed to various sublethal concentrations of copper(I) chloride (LC₅₀ = 281 μM, i.e., 17.8 mg Cu L⁻¹) and Irgarol 1051 (LC₅₀ > 500 μM, i.e., >127 mg L⁻¹), we evaluated their immunotoxic effects through a series of cytochemical assays previously used for organotin compounds. Both compounds can induce dose-dependent immunosuppression, acting on different cellular targets and altering many activities of immunocytes but, unlike TBT, did not have significant effects on cell morphology. Generally, Cu(I) appeared to be more toxic than Irgarol 1051: it significantly (p < 0.05) inhibited yeast phagocytosis at 0.1 μM (∼10 μg L⁻¹), and affected calcium homeostasis and mitochondrial cytochrome-c oxidase activity at 0.01 μM (∼1 μg L⁻¹). Both substances were able to change membrane permeability, induce apoptosis from concentrations of 0.1 μM (∼10 μg L⁻¹) and 200 μM (∼50 mg L⁻¹) for Cu(I) and Irgarol 1051, respectively, and alter the activity of hydrolases. Both Cu(I) and Irgarol 1051 inhibited the activity of phenoloxidase, but did not show any interactive effect when co-present in the exposure medium, suggesting different mechanisms of action.     

Distribution of Se and its species in Myriophyllum spicatum and Ceratophyllum demersum growing in water containing Se (VI)

The uptake of Se (VI) by two aquatic plants, Myriophyllum spicatum L. and Ceratophyllum demersum L., and its effects on their physiological characteristics have been studied. Plants were cultivated outdoors under semi-controlled conditions and in two concentrations of Na selenate solution (20 μg Se L⁻¹ and 10 mg Se L⁻¹). The higher dose of Se reduced the photochemical efficiency of PSII in both species, while the lower dose had no effect on PSII. Addition of Se had no effect on the amounts of chlorophyll a and b. The concentration of Se in plants grown in 10 mg Se L⁻¹, averaged 212 ± 12 μg Se g⁻¹ DM in M. spicatum (grown from 8-13 d), and 492 ± 85 μg Se g⁻¹ DM in C. demersum (grown for 31 d). Both species could take up a large amount of Se. The amount of soluble Se compounds in enzyme extracts ranged from 16% to 26% in control, and in high Se solution from 48% to 36% in M. spicatum and C. demersum, respectively. Se-species were determined using HPLC-ICP-MS. The main soluble species in both plants was selenate (∼37%), while SeMet and SeMeSeCys were detected at trace levels.     

Implication of organic acids in the long-distance transport and the accumulation of lead in Sesuvium portulacastrum and Brassica juncea

The implication of organic acids in Pb translocation was studied in two species varying in shoot lead accumulation, Sesuvium portulacastrum and Brassica juncea. Citric, fumaric, malic and α-cetoglutaric acids were separated and determined by HPLC technique in shoots, roots and xylem saps of the both species grown in nutrient solutions added with 200 and 400 μM of Pb(II). The lead content of the xylem saps was determined by ICP–MS. Results showed that S. portulacastrum is more tolerant to Pb than B. juncea. Lead concentration in xylem sap of the S. portulacastrum was significantly greater than in that of B. juncea. For both species, a positive correlation was established between lead and citrate concentrations in xylem sap. However minor relationship was observed for fumaric, malic and α-cetoglutaric acids. In the shoots lead treatment also induced a significant increase in citric acid concentration. Both observations suggest the implication of citric acid in lead translocation and shoot accumulation in S. portulacastrum and B. juncea. The relatively high accumulation of citric acid in xylem sap and shoot of S. portulacastrum could explain its high potential to translocate and accumulate this metal in shoot suggesting their possible use to remediate Pb polluted soils.     

Investigation of trace metal binding properties of lignin by diffusive gradients in thin films

The binding behavior of lignin for Pb, Cu, Co, Mn, Cd and Ni was studied using the diffusive gradients in thin films technique (DGT). Samplers with different structures of diffusive gel were used in the well-stirred systems containing known concentrations of metals along with (a) 10, 20 and 40 μM lignin and; (b) 0.64 and 6.47 μM Suwannee river fulvic acid + 40 μM lignin at an ionic strength of 0.01 M (NaNO₃) and pH = 7. Diffusion coefficients of lignin complexes in acrylamide gels were estimated and found to be less than 5% of the equivalent coefficients for the uncomplexed metal ions. These values were used to calculate concentrations of labile metals from DGT measurements in solutions, where lignin could discriminate metals in the order of Pb⁺² > Cu⁺² > Cd⁺² > Ni⁺² > Co⁺² > Mn⁺². Stability constants (Log K) were calculated using Visual MINTEQ II and WHAM V software. The K values were compared with the stability constants from titration of Pb and Cd with 10 μM lignin aqueous samples and with those of humic substances in natural waters. The constants obtained from measurement of complexing capacities might bias the real corresponding values unless two line regression analyses on titration data are considered. The DGT study of fractionation of metal species at varying ratios indicated that the proportion of organic complexes decreased with increasing ratios and gradually more metals were exchanged with inorganic phases. Speciation of Pb and Cd is affected by the concentrations of FA, Cd is dominantly bound with FA while Pb is evenly partitioned between the ligands. The comprehensive knowledge of metal-lignin complexes sheds some light on in situ operational speciation information that can be achieved by DGT.     

Biphasic effects of lanthanum on Vicia faba L. seedlings under cadmium stress, implicating finite antioxidation and potential ecological risk

In the present study, lanthanum (La) as a representative REE was used to explore the mechanisms for alleviation of Cd-induced oxidative damage by extraneous La at appropriate concentrations, and to assess ecological risk of combination of Cd and La at higher concentrations in roots of Vicia faba L. seedlings. The seedlings were hydroponically cultured for 15 d under nutrient solution, 6 μmol L⁻¹ CdCl₂, and combination of 6 μmol L⁻¹ CdCl₂ and increasing concentrations of La, respectively. The results showed that the supplementation with low concentrations of exogenous La (<120 μmol L⁻¹) led to reduced contents of Cd, Ca, Cu, Zn, Mn or Fe element and increased activities of superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (GPX) and ascorbate peroxidase (APX) isozymes as well as heat shock protein 70 (HSP 70) production in the roots. However, the supplementation with higher La (>120 μmol L⁻¹) showed the adverse effects. The contents of Cd elevated above the single Cd treatment in the roots, accompanying with the decline of antioxidant isozyme’s activities and HSP 70, and increment of carbonylated proteins and endoprotease isozyme’s activities. The results also showed that the root growth was not only related to carbonylated proteins, but also to indole acetic acid oxidase activities. Therefore, the supplemented extraneous La contributed to biphasic effects: stimulated antioxidation at lower concentrations and pro-oxidation at higher concentrations against Cd-induced oxidative stress in the roots.     

Comparison of EDTA-enhanced phytoextraction and phytostabilisation strategies with Lolium perenne on a heavy metal contaminated soil

Phytoremediation is a promising and cost-effective strategy to manage heavy metal polluted sites. In this experiment, we compared simultaneously phytoextraction and phytostabilisation techniques on a Cd and Zn contaminated soil, through monitoring of plant accumulation and leaching. Lolium perenne plants were cultivated for 2 months under controlled environmental conditions in a 27.6 dm³-pot experiment allowing the collect of leachates. The heavy metal phytoextraction was promoted by adding Na-EDTA (0.5 g kg⁻¹ of soil) in watering solution. Phytostabilisation was assessed by mixing soil with steel shots (1%) before L. perenne sowing. Presence of plants exacerbated heavy metal leaching, by improving soil hydraulic conductivity. Use of EDTA for phytoextraction led to higher concentration of heavy metal in shoots. However, this higher heavy metal extraction was insufficient to satisfactory reduce the heavy metal content in soil, and led to important heavy metal leaching induced by EDTA. On the other hand, addition of steel shots efficiently decreased both Cd and Zn mobility, according to 0.01 M CaCl₂ extraction, and leaching. However, improvement of growth conditions by steel shots led to higher heavy metal mass in shoot tissues. Therefore, soil heavy metal mobility and plant metal uptake are not systematically positively correlated.     

Chelant-aided enhancement of lead mobilization in residential soils

Chelation of metals is an important factor in enhancing solubility and hence, availability to plants to promote phytoremediation. We compared the effects of two chelants, namely, ethylenediaminetetraacetic acid (EDTA) and ethylenediaminedisuccinic acid (EDDS) in enhancing mobilized lead (Pb) in Pb-based paint contaminated residential soils collected from San Antonio, Texas and Baltimore, Maryland. Batch incubation studies were performed to investigate the effectiveness of the two chelants in enhancing mobilized Pb, at various concentrations and treatment durations. Over a period of 1 month, the mobilized Pb pool in the San Antonio study soils increased from 52 mg kg⁻¹ to 287 and 114 mg kg⁻¹ in the presence of 15 mM kg⁻¹ EDTA and EDDS, respectively. Stepwise linear regression analysis demonstrated that pH and organic matter content significantly affected the mobilized Pb fraction. The regression models explained a large percentage, from 83 to 99%, of the total variation in mobilized Pb concentrations.     

Hydrolysis of di-n-butyl phthalate, butylbenzyl phthalate and di(2-ethylhexyl) phthalate in human liver microsomes

Diester phthalates are industrial chemicals used primarily as plasticizers to import flexibility to polyvinylchloride plastics. In this study, we examined the hydrolysis of di-n-butyl phthalate (DBP), butylbenzyl phthalate (BBzP) and di(2-ethylhexyl) phthalate (DEHP) in human liver microsomes. These diester phthalates were hydrolyzed to monoester phthalates (mono-n-butyl phthalate (MBP) from DBP, mono-n-butyl phthalate (MBP) and monobenzyl phthalate (MBzP) from BBzP, and mono(2-ethylhexyl) phthalate (MEHP)) by human liver microsomes. DBP, BBzP and DEHP hydrolysis showed sigmoidal kinetics in V-[S] plots, and the Hill coefficient (n) ranged 1.37-1.96. The S₅₀, V_max and CL_max values for DBP hydrolysis to MBP were 99.7 μM, 17.2 nmol min⁻¹ mg⁻¹ protein and 85.6 μL min⁻¹ mg⁻¹ protein, respectively. In BBzP hydrolysis, the values of S₅₀ (71.7 μM), V_max (13.0 nmol min⁻¹ mg⁻¹ protein) and CL_max (91.3 μL min⁻¹ mg⁻¹ protein) for MBzP formation were comparable to those of DBP hydrolysis. Although the S₅₀ value for MBP formation was comparable to that of MBzP formation, the V_max and CL_max values were markedly lower (<3%) than those for MBzP formation. The S₅₀, V_max and CL_max values for DEHP hydrolysis were 8.40 μM, 0.43 nmol min⁻¹ mg⁻¹ protein and 27.5 μL min⁻¹ mg⁻¹ protein, respectively. The S₅₀ value was about 10% of DBP and BBzP hydrolysis, and the V_max value was also markedly lower (<3%) than those for DBP hydrolysis and MBzP formation for BBzP hydrolysis. The ranking order of CL_max values for monoester phthalate formation in DBP, BBzP and DEHP hydrolysis was BBzP to MBzP ? DBP to MBP > DEHP to MEHP > BBzP to MBP. These findings suggest that the hydrolysis activities of diester phthalates by human liver microsomes depend on the chemical structure, and that the metabolism profile may relate to diester phthalate toxicities, such as hormone disruption and reproductive effects.     

Bioconcentration of triclosan, methyl-triclosan, and triclocarban in the plants and sediments of a constructed wetland

Constructed wetlands are a potential method for the removal of two pharmaceutical and personal care products from wastewater effluent. Triclosan (TCS; 5-chloro-2-[2,4-dichlorophenoxy]phenol) and triclocarban (TCC; 3,4,4′-trichlorocarbanillide) are antimicrobial agents added to a variety of consumer products whose accumulation patterns in constructed wetlands are poorly understood. Here, we report the accumulation of TCS, its metabolite methyl-triclosan (MTCS; 5-chloro-2-[2,4-dichlorophenoxy]), and TCC in wetland plant tissues and sediments. Three wetland macrophytes: Typha latifolia, Pontederia cordata, and Sagittaria graminea were sampled from a constructed wetland in Denton, Texas, USA. MTCS concentrations were below the method detection limit (MDL) for all species. TCS root tissue concentrations in T. latifolia were significantly greater than root concentrations in P. cordata (mean ± SE in ng g⁻¹: 40.3 ± 11.3 vs. 15.0 ± 1.9, respectively), while for TCC, shoot tissue concentrations in S. graminea were significantly greater than in T. latifolia (22.8 ± 9.3 vs. 9.0 (MDL), respectively). For both TCS and TCC, T. latifolia root tissue concentrations were significantly greater than shoot concentrations (TCS: 40.3 ± 11.3 vs. 17.2 ± 0.2, TCC: 26.0 ± 3.6 vs. 9.0, (MDL)). TCC concentrations in P. cordata roots were significantly greater than in shoots (34.4 ± 5.3 vs. 15.4 ± 2.8, respectively). TCS concentrations in T. latifolia roots and sediments and TCC concentrations in sediments generally decreased from wetland inflow to outflow. To our knowledge, this is the first study documenting species and tissue specific differences in the accumulation of TCS and TCC in plants from an operational constructed wetland. The species specific differences in bioaccumulation suggest TCS and TCC removal from constructed wetlands could be enhanced through targeted plantings.     

Estimation of daily intake of potentially toxic elements from urban street dust and the role of oral bioaccessibility testing

The pseudo-total and oral bioaccessible concentration of six potentially toxic elements (PTEs) in urban street dust was investigated. Typical pseudo-total concentrations across the sampling sites ranged from 4.4 to 8.6 mg kg⁻¹ for As, 0.2-3.6 mg kg⁻¹ for Cd, 25-217 mg kg⁻¹ for Cu, 14-46 mg kg⁻¹ for Ni, 70-4261 mg kg⁻¹ for Pb, and, 111-652 mg kg⁻¹ for Zn. This data compared favourably with other urban street dust samples collected and analysed in a variety of cities globally; the exception was the high level of Pb determined in a specific sample in this study. The oral bioaccessibility of PTEs in street dust is also assessed using in vitro gastrointestinal extraction (Unified Bioaccessibility Method, UBM). Based on a worst case scenario the oral bioaccessibility data estimated that Cd and Zn had the highest % bioaccessible fractions (median >45%) while the other PTEs i.e. As, Cu, Ni and Pb had lower % bioaccessible fractions (median <35%). The pseudo-total and bioaccessible concentrations of PTEs in the samples has been compared to estimated tolerable daily intake values based on unintentional soil/dust consumption. Cadmium, Cu and Ni are well within the oral tolerable daily intake rates. With respect to As and Pb, only the latter exceeds the TDI_oral if we model ingestion rate based on atmospheric ‘dustiness’ rather than the US EPA (2008) unintentional soil/dust consumption rate of 100 mg d⁻¹. We consider it unlikely that even a child with pica tendencies would ingest as much as 100 mg soil/dust during a daily visit to the city centre, and in particular to the sites with elevated Pb concentrations observed in this study.     

Analysis of phytochelatin complexes in the lead tolerant vetiver grass [Vetiveria zizanioides (L.)] using liquid chromatography and mass spectrometry

Ethylenediamene tetraacetic acid (EDTA) has been used to mobilize soil lead (Pb) and enhance plant uptake for phytoremediation. Chelant bound Pb is considered less toxic compared to free Pb ions and hence might induce less stress on plants. Characterization of possible Pb complexes with phytochelatins (PC_n, metal-binding peptides) and EDTA in plant tissues will enhance our understanding of Pb tolerance mechanisms. In a previous study, we showed that vetiver grass (Vetiveria zizanioides L.) can accumulate up to 19,800 and 3350 mg Pb kg⁻¹ dry weight in root and shoot tissues, respectively; in a hydroponics set-up. Following the basic incubation study, a greenhouse experiment was conducted to elucidate the efficiency of vetiver grass (with or without EDTA) in remediating Pb-contaminated soils from actual residential sites where Pb-based paints were used. The levels of total thiols, PC_n, and catalase (an antioxidant enzyme) were measured in vetiver root and shoot following chelant-assisted phytostabilization. In the presence of 15 mM kg ⁻¹ EDTA, vetiver accumulated 4460 and 480 mg Pb kg⁻¹ dry root and shoot tissue, respectively; that are 15- and 24-fold higher compared to those in untreated controls. Despite higher Pb concentrations in the plant tissues, the amount of total thiols and catalase activity in EDTA treated vetiver tissues was comparable to chelant unamended controls, indicating lowered Pb toxicity by chelation with EDTA. The identification of glutathione (referred as PC₁) (m/z 308.2), along with chelated complexes like Pb-EDTA (m/z 498.8) and PC₁-Pb-EDTA (m/z 805.3) in vetiver root tissue using electrospray tandem mass spectrometry (ES-MS) highlights the possible role of such species towards Pb tolerance in vetiver grass.     

Arsenite toxicity and uptake rate of rice (Oryza sativa L.) in vivo

Toxicity threshold of arsenite on intact rice seedlings was determined and arsenite uptake characteristics were investigated using non-toxic concentrations of arsenite.The arsenite toxicity threshold was 2.4 μM arsenite which reduced growth by 10% (EC₁₀). The two highest arsenite levels induced wilting of seedlings and reduced both, transpiration rate and net photosynthetic rate. Arsenic content in plant tissue increased up to 10.7 μM arsenite and then declined with increasing arsenite concentration in the treatment solution. The contents of Si, P, K, and of micronutrients Cu, Fe, Mn and Zn in shoot d.m. were reduced by arsenite levels ≥ 5.3 μM. In the non-toxic range, arsenite uptake rate was linearly related to arsenite concentration.High arsenite levels reduced growth without being taken up which might be due to increasing binding of arsenite to proteins at the outer side of the plasmalemma.