Toxicity of three halogenated flame retardants to nitrifying bacteria, red clover (Trifolium pratense), and a soil invertebrate (Enchytraeus crypticus)

Halogenated flame retardants have a high sorption affinity to particles, making soils and sediments important sinks. Here, three of the most commonly used flame retardants have been tested for sub-lethal toxicity towards soil nitrifying bacteria, a terrestrial plant (seed emergence and growth of the red clover, Trifolium pratense), and a soil invertebrate (survival and reproduction of Enchytraeus crypticus). Tetrabromobisphenol A (TBBPA) was quite toxic to enchytraeids, with significant effects on reproduction detected already at the 10 mgkg(-1) exposure level (EC(10)=2.7 mgkg(-1)). In contrast, decabromodiphenyl ether (DeBDE) was not toxic at all, and short-chain chloroparaffins (CP(10-13)) only affected soil nitrifying bacteria at the highest test concentration (EC(10)=570 mgkg(-1)). Exposure concentrations were verified by chemical analysis for TBBPA and DeBDE, but not for CP(10-13), as a reliable method was not available. Based on the generated data, a PNEC for soil organisms can be estimated at 0.3 mgkg(-1) for TBBPA and 57 mgkg(-1) for short-chain chloroparaffins. No PNEC could be estimated for DeBDE. Measurements of TBBPA in soil are not available, but measured concentrations in Swedish sludge are all lower than the estimated threshold value for biological effects in soil.     

Fate of tetrabromobisphenol A and hexabromocyclododecane brominated flame retardants in soil and uptake by plants

The fate of tetrabromobisphenol A (TBBPA) and hexabromocyclododecane diastereomers (α-, β-, and γ-HBCD) and uptake by plants (cabbage and radish) was investigated. In a short-term (8 weeks) experiment, sorption to soil matrix resulted in 90% decline in recovery of these compounds in the experimental soil. However, nearly 50% of initial HBCDs recovered in mixed cabbage-radish treatments, which suggested that interspecific plant interactions might enhance the bioavailability of HBCDs. Although both plant species could uptake TBBPA and HBCDs, cabbage showed greater accumulating ability. Up to 3.5-10.0-fold higher HBCD concentrations were observed than TBBPA concentrations in all plant tissues, and the distribution of HBCDs in plant tissues was diastereomer-specific. The predominance of α-HBCD in shoot tissues for both species might be attributed to diastereomer-specific translocation of HBCDs, shift in diastereomer pattern and/or selective metabolization of γ-HBCD within plants. The results showed that strong sorption to soil particles reduced the potential of human exposure to BFRs in the soil. However, plants increased the exposure risk by uptaking these compounds and by enhancing their bioavailability. The results also provide insight into transport mechanisms of TBBPA and HBCD diastereomers in soil-plant systems.     

Effects of tetrabromobisphenol A as an emerging pollutant on wheat (Triticum aestivum) at biochemical levels

Biochemical responses of wheat (Triticum aestivum) to the stress of tetrabromobisphenol A (TBBPA) as an emerging pollutant were examined. The results indicated that reduction of the chlorophyll (CHL) content in wheat leaves could be observed. However, the changes in the CHL content with the increasing TBBPA concentration from 50 to 5000 mg kg(-1) were insignificant (p>0.05). Increased malondialdehyde levels were detected in wheat leaves after both 7-d and 12-d exposures. The changes in the activity of superoxide dismutases (SOD), peroxidases (POD) and catalases (CAT) in wheat leaves irregularly fluctuated with time as the TBBPA concentration increased. However, significant (p<0.05) decrease in the activity of POD and CAT treated with 500 and 5000 mg kg(-1) TBBPA could be observed. Our data also showed that the plant has the capacity to tolerate the oxidative stress, but the capacity would be lost with prolonged exposure and increasing TBBPA concentration. There were no dose-response effects in the changes between the activity of antioxidant enzymes (SOD, POD and CAT) and the concentration of TBBPA. The decrease in the activity of POD and CAT could be considered as good biomarkers of serious stress by TBBPA in soil environment.     

Translocation of metals and its effects in the tomato plants grown on various amendments of tannery waste: evidence for involvement of antioxidants

A field experiment was conducted to study the effect of different amendments of tannery sludge on physiological and biochemical parameters of tomato plant (Lycopersicon esculentum L. Mill). The accumulation of metals (Cr, Fe) in different parts of tomato plants grown on tannery sludge amended soil increased in a concentration and duration-dependent manner. The accumulation of both the metals was found lowest in the fruits of the plant. The statistical analysis of the results showed an increase in chlorophyll and protein contents in lower sludge amendment ratio at all exposures followed by a decrease at highest (100%) sludge amendment ratio. Lipid peroxidation enhanced in both root and leaves of sludge grown plants of tomato at all the sludge amendments and exposure periods, which is evidenced by increased malondialdehyde content, however the maximum increase was found in the roots (43.63%) and leaves (56.66%) of the plant grown on 100% tannery sludge at 60 d, over respective controls. The level of antioxidants, cysteine, non-protein thiol and ascorbic acid increased in the sludge grown plants of tomato to cope up with stress induced by the excess amount of the heavy metals present in the tannery sludge. The maximum increase was found in cysteine content (75.53% in the leaves), non-protein thiol content (92.68% in the roots) and ascorbic acid content (29.66% in the roots) of the plant at 75% tannery sludge after 30 d. The tomato plants were found well adopted for minimizing damage induced by reactive oxygen species, when grown on tannery sludge amendments in the present study.     

Mercury in salt marshes ecosystems: Halimione portulacoides as biomonitor

Mercury concentrations were quantified in Halimione portulacoides (roots, stems and leaves) as well as in sediments from eight Portuguese estuarine systems, covering seventeen salt marshes with distinct degrees of mercury contamination. The concentration of mercury in the sediments ranged from 0.03 to 17.0 microg g(-1). The results show that the accumulation of mercury differed according to the organ of the plant examined and the concentration of mercury in the sediments. Higher mercury concentrations were found in the roots (up to 12.9 microg g(-1)) followed by the leaves (up to 0.12 microg g(-1)), while the stems had the lowest concentrations (up to 0.056 microg g(-1)). A linear model explained the relation between the concentrations of mercury in the different plant organs: roots and stems (R(adj)(2)=0.75), stems and leaves (R(adj)(2)=0.85) and roots and leaves (R(adj)(2)=0.78). However, the results show that the variation of mercury concentration in the roots versus mercury concentration in the sediments was best fitted by a sigmoidal model (R(adj)(2)=0.89). Mercury accumulation in the roots can be described in three steps: at a low range of mercury concentrations in the sediments (from 0.03 up to 2 microg g(-1)), the accumulation of mercury in roots is also low reaching a maximum concentration of 1.3 microg g(-1); the highest rates of mercury accumulation in the roots occur in a second step, until the concentrations of mercury in the sediments reach approximately 4.5 microg g(-1); after reaching this maximum value, the rate of mercury accumulation in the roots slows down leading to a plateau in the concentration of mercury in the roots of about 9.4 microg g(-1), which corresponds to a mercury concentration in the sediments of about 11 microg g(-1). A linear model explained also the accumulation of mercury in leaves versus the mercury concentration in the sediments (R(adj)(2)=0.88). Differences in responses of roots and leaves are explained by the dynamics of the plant organs: old roots are mineralised in situ close to new roots, while leaves are renewed. Previous studies have already shown that H. portulacoides is a bioindicator for mercury and the results from this work sustain that H. portulacoides may also be used as a biomonitor for mercury contamination in salt marshes. Nevertheless, caution should be taken in the application of the models, concerning the life cycle of the species and the spatial variability of the systems.     

Lead detoxification by coontail (Ceratophyllum demersum L.) involves induction of phytochelatins and antioxidant system in response to its accumulation

Coontail (Ceratophyllum demersum L.) plants when exposed to various concentrations of Pb (1-100microM) for 1-7days, exhibited both phytotoxic and tolerance responses. The specific responses were function of concentration and duration. Plants accumulated 1748mugPbg(-1) dw after 7d which reflected its metal accumulation ability, however most of the metal (1222microgg(-1) dw, 70%) was accumulated after 1d exposure only. The toxic effect and oxidative stress caused by Pb were evident by the reduction in biomass and photosynthetic pigments and increase in malondialddehyde (MDA) content and electrical conductivity with increase in metal concentration and exposure duration. Morphological symptoms of senescence phenomena such as chlorosis and fragmentation of leaves were observed after 7d. The metal tolerance and detoxification strategy adopted by the plant was investigated with reference to antioxidant system and synthesis of phytochelatins. Protein and antioxidant enzymes viz., superoxide dismutase (SOD, EC 1.15.1.1), guaiacol peroxidase (GPX, EC 1.11.1.7) ascorbate peroxidase (APX, EC 1.11.1.11), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) showed induction at lower concentration and duration followed by decline. All enzymes except GPX showed maximum activity after 1d. An increase in cysteine, non-protein thiols (NP-SH) and glutathione (GSH) content was observed at moderate exposure conditions followed by decline. Phytochelatins (PC(2) and PC(3)) were synthesized to significant levels at 10 and 50microM Pb with concomitant decrease in GSH levels. Thus production of PCs seems important for the detoxification of metal, however it may lead to depletion of GSH and consequently oxidative stress. Results suggest that plants responded positively to moderate Pb concentrations and accumulated high amount of metal. Due to metal accumulation coupled with detoxification potential, the plant appears to have potential for its use as phytoremediator species in aquatic environments having moderate pollution of Pb.     

Translocation of metals from fly ash amended soil in the plant of Sesbania cannabina L. Ritz: effect on antioxidants

The plants of Sesbania cannabina Ritz grown on different amendments of fly ash (FA), have shown a high accumulation of metals (Fe, Mn, Zn, Cu, Pb and Ni). The highest accumulation of Fe the and lowest level of Ni were recorded in these plants. The different amendments of fly ash with garden soil (GS) were extracted with DTPA and the levels of metals were found to be decreased with an increase in fly ash application ratio from 10% to 50% FA. The analysis of the results showed an increase in the level of malondialdehyde (MDA) content of the roots for all the exposure periods. The maximum increases of 136% (roots) and 120% (leaves) were observed in MDA content at 100% FA after 30 d of growth of the plant, compared to GS. The level of antioxidants was found to increase for all the exposure periods in the roots of the plants to combat metal stress. At 30 d, the maximum increase of 57% (ascorbic acid) and 78% (free proline) was observed in the roots of the plants grown on 100% and 10% FA, respectively, as compared to their respective GS. At 90 d, a maximum increase of 42% (cysteine) and 117% (NPSH) was recorded in the roots of the plants grown on 25% and 100% FA, respectively, as compared to their respective GS. In leaves, a significant increase in antioxidants i.e. cysteine, NPSH and free proline content was recorded after 30 d, whereas no such trend was observed for the rest of the exposure periods. The chlorophyll and carotenoid contents increased with an increase in the FA amendment ratio from 10% to 50% FA for all the exposure periods as compared to GS. In both roots and leaves, the level of protein content increased in all the amendments and 100% FA at 30 d as compared to GS. Thus, there is a balance in the level of MDA content and level of antioxidants in the plants at 90 d. In view of its tolerance, the plants may be used for phytoremediation of metals from fly ash contaminated sites and suitable species for plantation on fly ash land fills.     

Chromium induced lipid peroxidation in the plants of Pistia stratiotes L.: role of antioxidants and antioxidant enzymes

In the plant, Pistia stratiotes L., the effect of different concentrations of chromium (0, 10, 40, 80 and 160 microM) applied for 48, 96 and 144 h was assessed by measuring changes in the chlorophyll, protein, malondialdehyde (MDA), cysteine, non-protein thiol, ascorbic acid contents and superoxide dismutase (SOD), ascorbate peroxidase (APX) and guiacol peroxidase (GPX) activity of the plants. Both in roots and leaves, an increase in MDA content was observed with increase in metal concentration and exposure periods. In roots, the activity of antioxidant enzymes viz. SOD and APX increased at all the concentrations of Cr at 144 h than their controls. The GPX activity of the treated roots increased with increase in Cr concentration at 48 and 96 h of exposures, however, at 144 h its activity was found declined beyond 10 microM Cr. The level of antioxidants in the roots of the treated plant viz. cysteine and ascorbic acid was also found increased at all the concentrations of Cr at 144 h than their respective controls, however, an increase in the non-protein thiol content was recorded up to 40 microM Cr followed by decrease. The chlorophyll content decreased with increase in Cr concentrations and exposure periods. However, the protein content of both roots and leaves were found decreased with increase in Cr concentrations at all the exposure periods except an increase was recorded at 10 microM Cr at 48 h. In Cr treated plants, the no observed effect concentration (NOEC) and lowest observed effect concentration (LOEC) for leaves chlorophyll and protein contents were 40 and 80 microM Cr, respectively after 48 h exposure while NOEC and LOEC for root protein content were 10 and 40 microM, respectively after 48 h. The analysis of correlation coefficient data revealed that the metal accumulation in the roots of the plant was found positively correlated with antioxidant parameters except SOD after 48 h of exposure, however, negatively correlated with most of all the parameters studied at 144 h in both part of the plant. It may be suggested from the present study that toxic concentrations of Cr cause oxidative damage as evidenced by increased lipid peroxidation and decreased chlorophyll and protein contents. However, the higher levels of enzymatic and non-enzymatic antioxidants suggest the reason for tolerating higher levels of metals.     

Response of antioxidants in sunflower (Helianthus annuus L.) grown on different amendments of tannery sludge: its metal accumulation potential

The interaction of metals present in tannery waste and their tolerance in the plants of sunflower (Helianthus annuus L.) was studied in the present paper under field conditions. Effects of 100% tannery sludge and various amendments of tannery sludge (10%, 25%, 35%, 50%, 75%) along with one set of control were studied on the physiological and biochemical parameters of the plant along with their metal accumulation potential after 30, 60 and 90 d after sowing. The plants of H. annuus were found effective in the accumulation of metals (Cr, Fe, Zn and Mn) in roots, shoots and leaves, however, the level of toxic metal, Cr was found below detection limit in the seeds of the plant. The oil was extracted from the seeds of the plant and the level of oil content was increased up to 35% tannery sludge as compared to control followed by decrease at higher tannery sludge ratio. An increase in the chlorophyll, protein, cysteine, non-protein thiol and sugar contents was observed at the lower amendment of tannery sludge at initial exposure periods followed by decrease than their respective controls. Malondialdehyde content in the roots and leaves was increased beyond 50% sludge amendments at all the exposure periods as compared to control. However, proline and ascorbic acid contents of the roots and leaves of the plant increased at all the exposure periods and sludge amendments, compared to their respective controls.     

Sequestration of maize crop straw C in different soils: role of oxyhydrates in chemical binding and stabilization as recalcitrance

While biophysical controls on the sequestration capacity of soils have been well addressed with physical protection, chemical binding and stabilization processes as well as microbial community changes, the role of chemical binding and stabilization has not yet well characterized for soil organic carbon (SOC) sequestration in rice paddies. In this study, a 6-month laboratory incubation with and without maize straw amendment (MSA) was conducted using topsoil samples from soils with different clay mineralogy and free oxy-hydrate contents collected across Southern China. The increase in SOC under MSA was found coincident with that in Fe- and Al-bound OC (Fe/Al-OC) after incubation for 30 d (R(2)=0.90, P=0.05), and with sodium dithionate-citrate-bicarbonate (DCB) extractable Fe after incubation for 180 d (R(2)=0.99, P<0.01). The increase in SOC under MSA was found higher in soils rich in DCB extractable Fe than those poor in DCB extractable Fe. The greater SOC sequestration in soils rich in DCB extractable Fe was further supported by the higher abundance of (13)C which was a natural signature of MSA. Moreover, a weak positive correlation of the increased SOC under MSA with the increased humin (R(2)=0.87, P=0.06) observed after incubation for 180 d may indicate a chemical stabilization of sequestered SOC as humin in the long run. These results improved our understanding of SOC sequestration in China's rice paddies that involves an initial chemical binding of amended C and a final stabilization as recalcitrant C of humin.     

Tetrabromobisphenol A: tissue distribution in fish, and seasonal variation in water and sediment of Lake Chaohu, China

Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardants (BFRs). It has been detected in various environment media and has been approved to be high toxic to aquatic organisms. However, the exposure levels of TBBPA in the main watersheds in China have not been investigated sufficiently. In this study TBBPA concentrations in water, sediment and tissues of four fish species from Lake Chaohu were determined. Spatial and temporal distribution patterns of TBBPA in water and sediment, and the relationship among TBBPA concentrations in fish tissues and fish size were analyzed. The results showed that the maximum TBBPA concentration in sediment was 518 ng/g in March 2008. It is almost the highest value than those ever reported anywhere in the world. In lake water, TBBPA level reached the maximum value of 4.87???g/L in July. Tissue distributions of TBBPA in four fish species were similar, and the mean concentrations in the fish were in the range of 28.5?C39.4 ng/g, much higher than those reported in Japan, Europe, and the United States. The maximum concentrations of TBBPA were found in kidneys where TBBPA concentration was positively correlated with fish size of Cyprinus carpio. Results demonstrated a widespread distribution of TBBPA in Lake Chaohu. The source was mainly inflow rivers near the cities. The distribution coefficient among water, fish, and sediment was 1:28:117. This indicated that sediment was the main repository for TBBPA within Lake Chaohu. Furthermore, TBBPA may pose a potential ecological risk in the lake during summer.     

Field assessment of oxytetracycline exposure to the freshwater macrophytes Egeria densa Planch. and Ceratophyllum demersum L

In a microcosm study, two aquatic macrophytes, Egeria densa and Ceratophyllum demersum were exposed to nominal concentrations of 0, 5, 20, 50, and 250 microg/L oxytetracycline (n=3), plus 20 microg/L oxytetracycline amended with additional nitrogen (N) and phosphorus (P). Responses were monitored bi-weekly over a six-week exposure period. Both plant species exhibited a significant decline in growth in the 250 microg/L oxytetracycline and the N- and P-amended units. Decreased light penetration resulting from accumulating oxytetracycline by-products appears to be the primary modifier in the growth of these plants. Increased susceptibility to oxytetracycline exposure was noted in some paired plantings (e.g., E. densa root development), relative to individual plants in these treatments, however, no clear explanation for this response is available. Based on the toxicity data generated in this study, we estimate that current concentrations of oxytetracycline in freshwater environments do not pose a direct risk to E. densa and C. demersum.     

Airborne foliar transfer of particular metals in Lactuca sativa L.: translocation,phytotoxicity, and bioaccessibility

The uptake, translocation, and human bioaccessibility of metals originating from atmospheric fine particulate matters (PM) after foliar exposure is not well understood. Lettuce (Lactuca sativa L.) plants were exposed to micronic PbO, CuO, and CdO particulate matters (PMs) by the foliar pathway and mature plants (6 weeks old) were analyzed in terms of: (1) metal accumulation and localization on plant leaf surface, and metal translocation factor (TF) and global enrichment factor (GEF) in the plants; (2) shoot growth, plant dry weight (DW), net photosynthesis (Pn), stomatal conductance (Gs), and fatty acid ratio; (3) metal bioaccessibility in the plants and soil; and (4) the hazard quotient (HQ) associated with consumption of contaminated plants. Substantial levels of metals were observed in the directly exposed edible leaves and newly formed leaves of lettuce, highlighting both the possible metal transfers throughout the plant and the potential for human exposure after plant ingestion. No significant changes were observed in plant biomass after exposure to PbO, CuO, and CdO-PMs. The Gs and fatty acid ratio were increased in leaves after metal exposure. A dilution effect after foliar uptake was suggested which could alleviate metal phytotoxicity to some degree. However, plant shoot growth and Pn were inhibited when the plants are exposed to PbO, and necrosis enriched with Cd was observed on the leaf surface. Gastric bioaccessibility of plant leaves is ranked: Cd?>?Cu?>?Pb. Our results highlight a serious health risk of PbO, CuO, and CdO-PMs associated with consumption of vegetables exposed to these metals, even in newly formed leaves in the case of PbO and CdO exposure. Finally, the study highlights the fate and toxicity of metal rich-PMs, especially in the highly populated urban areas which are increasingly cultivated to promote local food.

     

Cadmium accumulation and its influence on lipid peroxidation and antioxidative system in an aquatic plant, Bacopa monnieri L

Bacopa monnieri L. plants exposed to 10, 50, 100 and 200 microM cadmium (Cd) for 48, 96 and 144 h were analysed with reference to the accumulation of metal and its influence on various enzymatic and non-enzymatic antioxidants, thiobarbituric acid reactive substances (TBARS), photosynthetic pigments and protein content. The accumulation of Cd was found to be increased in a concentration and duration dependent manner with more Cd being accumulated in the root. TBARS content of the treated roots and leaves increased with increase in Cd concentration and exposure periods, indicating the occurrence of oxidative stress. Induction in the activities of superoxide dismutase (SOD), ascorbate peroxidase (APX) and guiacol peroxidase (GPX) was recorded in metal treated roots and leaves of B. monnieri. In contrast, a significant reduction in catalase activity in Cd treated B. monnieri was observed. An increase was also noted in the levels of cysteine and non-protein thiol contents of the roots of B. monnieri followed by a decline. However, in leaves, cysteine and non-protein thiol contents were found to be enhanced at all the Cd concentrations and exposure periods. A significant reduction in the level of ascorbic acid was observed in a concentration and duration dependent manner. The total chlorophyll and protein content of B. monnieri decreased with increase in Cd concentration at all the exposure periods. Results suggest that toxic concentrations of Cd caused oxidative damage as evidenced by increased lipid peroxidation and decreased chlorophyll and protein contents. However, B. monnieri is able to combat metal induced oxidative injury involving a mechanism of activation of various enzymatic and non-enzymatic antioxidants.     

Treatment of phenolic wastewater by horseradish peroxidase immobilized by bioaffinity layering

Horseradish peroxidase was immobilized by bioaffinity layering and used for the treatment of wastewater containing p-chlorophenol. For this purpose, lectin Concanavalin A was bound to Sephadex beads. The glycoenzyme peroxidase was layered upon this Con A layer. Subsequently, alternate layers of the enzyme and Con A were applied. The most efficient design consisted of three layers of Con A and peroxidase each. This immobilized enzyme preparation retained 80% of the activity of the free peroxidase used for immobilization. PEG at the concentration of 0.1 mg ml(-1) was found to prevent enzyme inactivation by the products, although it increased the process time. Thus 60 U ml(-1) of enzyme completely converted the p-chlorophenol (into products) in 4 min in the absence of PEG. On the other hand, only 0.05 U ml(-1) of enzyme was required for this purpose in the presence of PEG but the process required 60 min. Peroxidase converts phenol molecules into free radicals. These free radicals then polymerize and get precipitated. As a further means of minimizing exposure of the enzyme to free radicals and enhancing the reusability, it was decided to remove the enzyme from reaction medium after 10 min. With this strategy, the bioaffinity layered peroxidase preparation could be reused five times without any loss of activity.     

Effect of cadmium on free amino acid, glutathione and ascorbic acid concentrations in two barley genotypes (Hordeum vulgare L.) differing in cadmium tolerance

Hydroponic experiment was carried out to study the effect of three Cd levels on glutathione (GSH), free amino acids (FAA), and ascorbic acid (ASA) concentration in the different tissues of 2 barley cultivars with different Cd tolerance. Cadmium concentration in both roots and shoots increased with external Cd level, while biomass and ASA concentration declined, and Wumaoliuling, a Cd-sensitive genotype was more affected than ZAU 3, a Cd-tolerant genotype. The effect of Cd on GSH concentration was dose- and time-dependent. In the 5 d exposure, root GSH concentration increased in 0.5 microM Cd treatment compared with control, but decreased significantly in 5 microM Cd treatment, irrespective of genotypes. However, in the 10 d exposure, GSH concentration in all plant tissues decreased with increasing Cd levels in the culture medium, and Wumaoliuling was much more affected than ZAU 3. Cadmium treatment greatly altered FAA concentration and composition in plants. The effect of Cd on glutathione (Glu) concentration in roots varied with genotypes. ZAU 3 showed a steady increase in root Glu concentration in both 0.5 and 5 microM Cd treatments, while Wumaoliuling was decreased by 38.0% in 5 microM Cd treatment, compared with the control. The results indicate that GSH and ASA are attributed to Cd tolerance in barley plants, and the relative less reduction in GSH concentration in ZAU 3 under Cd stress relative to the control may account for its higher Cd tolerance.     

EDTA and urease effects on Hg accumulation by Lepidium sativum

The phytoextraction process was conducted under laboratory conditions with the use of garden cress plants (Lepidium sativum). The experiment was carried out in a model soil, which was characterized before conducting the process. Inorganic forms of mercury (HgCl(2), HgSO(4), Hg(NO(3))(2)) were used for contamination of the soil. The phytoextraction process was conducted after EDTA application to the soil and after urease application. Also the influence of simultaneous addition of ethylenediaminetetraacetic acid (EDTA) and urease into the soil on phytoextraction process was measured. In all variants of phytoextraction process the total mercury concentrations in roots, stems and leaves of garden cress were determined. The result showed that garden cress accumulated mercury from soil. The overall maximum concentration of mercury in its compounds was found in roots of the plant. In all cases, before addition of urease and EDTA, the translocation process and distribution of mercury in the plant tissues were limited. The addition of urease caused an increase of enzyme activity in the soil and at the same time caused an increase of mercury concentration in plant tissues. Application of EDTA increased solubility of mercury and caused an increase of metal accumulation by plants. After simultaneous addition of EDTA and urease into the soil garden cress accumulated about 20% of total mercury concentration in the soil. Most of mercury compounds were accumulated in leaves and stems of the plants (46.0-56.9% of total mercury concentration in the plant tissues).     

Timing of phosphate application affects arsenic phytoextraction by Pteris vittata L. of different ages

The effects of timing in phosphate application on plant growth and arsenic removal by arsenic hyperaccumulator Pteris vittata L. of different ages were evaluated. The hydroponic experiment consisted of three plant ages (A45d, A90d and A180d) and three P feeding regimens (P200+0, P134+66 and P66+134) growing for 45 d in 0.2-strength Hoagland-Arnon solution containing 145 microg L(-1) As. While all plants received 200 microM P, P was added in two phases: during acclimation and after arsenic exposure. High initial P-supply (P200+0) favored frond biomass production and plant P uptake, while split-P application (P134+66 and P66+134) favored plant root production. Single P addition favored arsenic accumulation in the roots while split-P addition increased frond arsenic accumulation. Young ferns (A45d) in treatment P134+66 were the most efficient in arsenic removal, reducing arsenic concentration to below 10 microg L(-1) in 35 d. The results indicated that the use of young ferns, coupled with feeding of low initial P or split-P application, increased the efficiency of arsenic removal by P. vittata.     

Cadmium accumulation and elimination in tissues of juvenile olive flounder, Paralichthys olivaceus after sub-chronic cadmium exposure

Experiments were carried out to investigate the accumulation and elimination of cadmium (Cd) in tissues (gill, intestine, kidney, liver and muscle) of juvenile olive flounder, Paralichthys olivaceus, exposed to sub-chronic concentrations (0, 10, 50, 100 microg l(-1)) of Cd. Cd exposure resulted in an increased Cd accumulation in tissues of flounder with exposure periods and concentration, and Cd accumulation in gill and liver increased linearly with the exposure time. At 20 days of Cd exposure, the order of Cd accumulation in organs was gill > intestine > liver > kidney > muscle and after 30 days of exposure, those were intestine > gill > liver > kidney > muscle. An inverse relationship was observed between the accumulation factor (AF) and the exposure level, but AF showed an increase with exposure time. During the depuration periods, Cd concentration in the gill, intestine and liver decreased immediately following the end of the exposure periods. No significant difference was found Cd in concentration in the kidney and muscle during depuration periods. The order of Cd elimination rate in organs were decreased intestine > liver > gill during depuration periods.     

The relationship between foliar injury, nitrogen metabolism, and growth parameters in ozonated soybeans

A single 12 h ozone exposure peaking at 0.20 ppm proved phytotoxic to greenhouse-grown 'Cutler 71' soybeans at each growth stage tested from V5 to R6. Visible injury occurred within 40 h on the unifoliodate leaves and middle-aged and older trifoliolates while the younger leaves were free from toxicity symptoms. In some instances visible injury was accompanied by a decrease in chlorophyll and an increase in leaf diffusive resistance. Although nitrogen fixation was not significantly altered except at early pod formation (R3), and nitrate reductase activity was significantly reduced only if the ozone exposure occurred at the time of maximal enzyme activity (V5), nitrogen content of the leaves was reduced by ozone treatment. Shoot dry weight was not affected 40 h after ozone treatment, but root dry weight was significantly reduced. Plants grown with supplemental NO(3)(-) were more sensitive to ozone than those dependent on fixed nitrogen. At plant maturity, there was no evidence of an ozone effect on shoot, root, or seed dry weight, NO(3)(-) -grown plants showed a significant increase in growth and yield over N(2)(-) plants; but no ozone effect was observed, despite the increased foliar sensitivity. Multiple ozone exposures at growth stages V3, R1 and R3 exacerbated the effects noted with a single episode and also reduced nitrogenase activity (reflected in specific and total nodule activity) and shoot and root dry weight. At plant maturity, there was again no evidence of a significant effect of multiple ozone treatment on shoot dry weight or seed yield although root weight remained low. The results would tend to support the hypothesis that older leaves of soybean do not make a significant contribution to seed yield. Although they may be injured by ozone during the reproductive phases of growth, seed yield may not be affected if the younger O(3)-tolerant leaves remain functional.