Toxicity of zinc oxide nanoparticles in the earthworm, Eisenia fetida and subcellular fractionation of Zn

The extensive use of nanoparticles (NPs) in a variety of applications has raised great concerns about their environmental fate and biological effects. This study examined the impact of dissolved organic matter (DOM) and salts on ZnO NP dispersion/solubility and toxicity to the earthworm Eisenia fetida. To be able to better evaluate the toxicity of NPs, exposure in agar and on filter paper was proposed for enabling a comparison of the importance of different uptake routes. A dose-related increase in mortality was observed in earthworms exposed in agar with almost 100% mortality after 96 h exposure to the highest concentration (1000 mg ZnO/kg agar). Scanning electron microscopy (SEM) showed that the addition of salts enhanced the aggregation of ZnO NPs in agar and consequently affected the dissolution behavior and biological availability of the particles. On filter paper, mortality was the highest at the lowest exposure concentration (50 mg ZnO/L) and seemed to decrease with increasing exposure levels. TEM images of ZnO showed that the solubility and morphology of NPs were changed dramatically upon the addition of humic acids (HA). The subcellular distribution pattern of Zn in earthworms after 96 h exposure in agar and on filter paper showed that the Zn taken up via dietary ZnO particles (from agar) was mainly found in organelles and the cytosol while the Zn accumulated as soluble Zn from filter paper was mainly distributed in cell membranes and tissues. Antioxidant enzymatic activities (SOD, CAT, and GSH-px) were investigated in the worms surviving the toxicity tests. A slight increase of SOD activities was observed at the lowest exposure dose of ZnO (50mg/kg), followed by a decrease at 100mg/kg in the agar cubes. Activities of both CAT and GSH-Px enzymes were not significantly influenced in the worms exposed to agar, although a slight decrease at 500 and 1000 mg ZnO/kg agar was observed. A similar change trend of SOD activities was observed for the earthworms on filter paper, but a significant decrease began at a higher ZnO NP concentration of 500 mg ZnO/L. The use of soil extracts instead of deionized water (DW) to simulate a realistic exposure system significantly reduced the toxicity of the ZnO NPs on filter paper, which increases the predictive power of filter paper toxicity tests for the environmental risk assessment of NPs.     

Influence of alumina coating on characteristics and effects of SiO2 nanoparticles in algal growth inhibition assays at various pH and organic matter contents

Silica nanoparticles (NPs) belong to the industrially most important NP types. In a previous study it was shown that amorphous SiO(2) NPs of 12.5 and 27.0 nm are stable in algal growth inhibition assays and that their ecotoxic effects are related to NP surface area. Here, it was hypothesized and demonstrated that an alumina coating completely alters the particle-particle, particle-test medium and particle-algae interactions of SiO(2) NPs. Therefore, stability and surface characteristics, dissolution, nutrient adsorption and effects on algal growth rate of both alumina coated SiO(2) NPs and bare SiO(2) NPs in OECD algal test medium as a function of pH (6.0-8.6) and natural organic matter (NOM) contents (0-12 mg C/l) were investigated. Alumina coated SiO(2) NPs aggregated in all media and adsorbed phosphate depending on pH and NOM concentration. On the other hand, no aggregation or nutrient adsorption was observed for the bare SiO(2) NPs. Due to their positive surface charge, alumina coated SiO(2) NPs agglomerated with Pseudokirchneriella subcapitata. Consequently, algal cell density measurements based on cell counts were unreliable and hence fluorescent detection of extracted chlorophyll was the preferred method. Alumina coated SiO(2) NPs showed lower toxicity than bare SiO(2) NPs at concentrations ≥46 mg/l, except at pH 6.0. At low concentrations, no clear pH effect was observed for alumina coated SiO(2) NPs, while at higher concentrations phosphate deficiency could have contributed to the higher toxicity of those particles at pH 6.0-6.8 compared to higher pH values. Bare SiO(2) NPs were not toxic at pH 6.0 up to 220 mg/l. Addition of NOM decreased toxicity of both particles. For SiO(2) NPs the 48 h 20% effect concentration of 21.8 mg/l increased 2.6-21 fold and a linear relationship was observed between NOM concentration and effective concentrations. No effect was observed for alumina coated SiO(2) NPs in presence of NOM up to 1000 mg/l. All experiments point out that the alumina coating completely altered NP interactions. Due to the difference in surface composition the SiO(2) NPs, which had the smallest surface area, were more toxic to the alga than the alumina coated SiO(2) NPs. Hence, surface modification can dominate the effect of surface area on toxicity.     

Neuronal cytotoxicity and genotoxicity induced by zinc oxide nanoparticles

Zinc oxide nanoparticles (ZnO NPs) are one of the most abundantly used nanomaterials in consumer products and biomedical applications. As a result, human exposure to these NPs is highly frequent and they have become an issue of concern to public health. Although toxicity of ZnO NPs has been extensively studied and they have been shown to affect many different cell types and animal systems, there is a significant lack of toxicological data for ZnO NPs on the nervous system, especially for human neuronal cells and tissues. In this study, the cytotoxic and genotoxic effects of ZnO NPs on human SHSY5Y neuronal cells were investigated under different exposure conditions. Results obtained by flow cytometry showed that ZnO NPs do not enter the neuronal cells, but their presence in the medium induced cytotoxicity, including viability decrease, apoptosis and cell cycle alterations, and genotoxicity, including micronuclei production, H2AX phosphorylation and DNA damage, both primary and oxidative, on human neuronal cells in a dose- and time-dependent manner. Free Zn^2 + ions released from the ZnO NPs were not responsible for the viability decrease, but their role on other types of cell damage cannot be ruled out. The results obtained in this work contribute to increase the knowledge on the genotoxic and cytotoxic potential of ZnO NPs in general, and specifically on human neuronal cells, but further investigations are required to understand the action mechanism underlying the cytotoxic and genotoxic effects observed.     

Silver nanoparticles: behaviour and effects in the aquatic environment

This review summarises and evaluates the present knowledge on the behaviour, the biological effects and the routes of uptake of silver nanoparticles (Ag NPs) to organisms, with considerations on the nanoparticle physicochemistry in the ecotoxicity testing systems used. Different types of Ag NP syntheses, characterisation techniques and predicted current and future concentrations in the environment are also outlined. Rapid progress in this area has been made over the last few years, but there is still a critical lack of understanding of the need for characterisation and synthesis in environmental and ecotoxicological studies. Concentration and form of nanomaterials in the environment are difficult to quantify and methodological progress is needed, although sophisticated exposure models show that predicted environmental concentrations (PECs) for Ag NPs in different environmental compartments are at the range of ng L(-1) to mg kg(-1). The ecotoxicological literature shows that concentrations of Ag NPs below the current and future PECs, as low as just a few ng L(-1), can affect prokaryotes, invertebrates and fish indicating a significant potential, though poorly characterised, risk to the environment. Mechanisms of toxicity are still poorly understood although it seems clear that in some cases nanoscale specific properties may cause biouptake and toxicity over and above that caused by the dissolved Ag ion. This review concludes with a set of recommendations for the advancement of understanding of the role of nanoscale silver in environmental and ecotoxicological research.     

Ecotoxicological effects of an aged TiO2 nanocomposite measured as apoptosis in the anecic earthworm Lumbricus terrestris after exposure through water, food and soil

Titanium dioxide nanoparticles seem to have a low toxicity to terrestrial organisms, though few studies are published in this area. TiO(2) used in sunscreens are nanocomposites where TiO(2) has been coated with magnesium, silica or alumina, as well as amphiphilic organics like polydimethyl siloxane (PDMS), and these coatings are modified by ageing. We assessed the ecotoxicity and propensity for bioaccumulation of an aged TiO(2) nanocomposite used in sunscreen cosmetics, and its potential effect on the frequency of apoptosis in different earthworm tissues. The earthworm Lumbricus terrestris was exposed to the TiO(2) nanocomposite for 7 days in water or 2-8 weeks in soil with the nanocomposite mixed either into food or soil at concentrations ranging from 0 to 100 mg kg(-1). Apoptosis was then measured by immunohistochemistry and Ti localized by XRF microscopy. Results showed no mortality, but an enhanced apoptotic frequency which was higher in the cuticule, intestinal epithelium and chloragogenous tissue than in the longitudinal and circular musculature. TiO(2) nanoparticles did not seem to cross the intestinal epithelium/chloragogenous matrix barrier to enter the coelomic liquid, or the cuticule barrier to reach the muscular layers. No bioaccumulation of TiO(2) nanocomposites could thus be observed.     

Environmental contamination of heavy metals from zinc smelting areas in Hezhang County, western Guizhou, China

Total heavy metal (Cd, Cr, Cu, Pb and Zn) concentrations were evaluated in smelting waste, soil, crop and moss samples collected from the Hezhang artisanal zinc smelting areas, Guizhou, China. Soil samples from the cornfield near the smelting sites contained extremely high Cd (5.8-74 mg kg(-1)), Pb (60-14,000 mg kg(-1)) and Zn (260-16,000 mg kg(-1)) concentrations. Elevated heavy metal concentrations were also found in corn plants and total Pb (0.80-1.5 mg kg(-1)) and Cd (0.05-0.76 mg kg(-1)) concentrations in corn grain have totally or partially exceeded the national guidance limits for foodstuff. Thus, the soil-to-crop transfer of heavy metals might pose a potential health risk to the local residents. Similar to the high heavy metal levels in soil and corn, Cd, Cr, Cu, Pb and Zn concentrations in moss samples collected from the smelting sites ranged from 10 to 110, 10 to 55, 26 to 51, 400 to 1200 and 330 to 1100 mg kg(-1), respectively, exhibiting a local spatial pattern of metals deposition from the atmosphere. Based on examination of Zn/Cd and Pb/Cd ratios of the analyzed samples, we have distinguished between the flue gas dust derived and smelting waste derived metals in different environmental compartments.     

Nonylphenol and octylphenol in human breast milk

Human milk is the most important form of nourishment for newborn children. Its consumption is strongly recommended by health authorities also for other important advantages. Unfortunately, in the last three decades a great number of investigations have shown the occurrence of several environmental contaminants in human milk, especially those with lipophilic properties. This study investigates the presence of nonylphenol, octylphenol (OP), nonylphenol monoethoxylate (NP1EO) and two octylphenol ethoxylates (OPEOs) (namely OP1EO and OP2EO), in human breast milk of Italian women. NP was the contaminant found at the highest levels with mean concentrations of 32 ng/mL, about two orders of magnitude higher than OP (0.08 ng/mL), OP1EO (0.07 ng/mL) and OP2EO (0.16 ng/mL). In the group of study a positive correlation among fish consumption and levels of NP in the milk was observed, in accordance with the evidence that seafood represents one of the most important sources of exposure to this group of contaminants in Italy. On the basis of the concentrations found in the breast milk samples, a maximum NP daily intake of 3.94 mug/kg/day can be calculated, which is close to the Tolerable Daily Intake (TDI) of 5 mug/kg body weight (bw) proposed by the Danish Institute of Safety and Toxicology. In the cases of OP no TDI is available, but its intake is at least six orders of magnitude lower than the NOAEL of 10 mg/kg/day derived from a two generation study on rats.     

湘潭锰矿重金属环境安全及植物耐性研究

采集了湘潭锰矿红旗分矿开采区、沙圹村恢复区的代表性当季蔬菜(莴笋叶Fruticicolidae、小白菜Brassica chinensis、香葱Allium schoenoprasum、空心菜Ipomoea aquatica)、废弃区的优势植物(商陆Phytolacca acinosa、野茼蒿Crassocephalum crepidioides、苍耳Xanthium sibiricum)和3个研究区的土壤,通过原子吸收分光光度法分析了Mn、Pb、Zn含量。结果表明:开采区蔬菜Mn含量(8.3~84.5 mg/kg)明显高于恢复区(2.7~55.6 mg/kg),开采区和恢复区蔬菜都明显受到Pb污染(0.6~33mg/kg),蔬菜Zn含量范围为1.9~6.5mg/kg;3个研究区域土壤重金属均明显超标,最严重的是Pb污染(1 993.5~2 213.5mg/kg)。商陆、野茼蒿和苍耳中重金属含量差异较大,对重金属的耐性强,其中商陆表现出最好的耐性与长势。研究结论对锰矿土地合理利用以及矿区土壤重金属治理提供一定的科学依据。     

Physiological effects of nanoparticles on fish: a comparison of nanometals versus metal ions

The use of nanoscale materials is growing exponentially, but there are also concerns about the environmental hazard to aquatic biota. Metal-containing engineered nanoparticles (NPs) are an important group of these new materials, and are often made of one metal (e.g., Cu-NPs and Ag-NPs), metal oxides (e.g., ZnO and TiO(2) NPs), or composite of several metals. The physiological effects and toxicity of trace metals in the traditional dissolved form are relatively well known and the overall aim of this review was to use our existing conceptual framework of metal toxicity in fish to compare and contrast the effects of nanometals. Conceptually, there are some fundamental differences that relate to bioavailability and uptake. The chemistry and behaviour of nanometals involves dynamic aspects of aggregation theory, rather than the equilibrium models traditionally used for free metal ions. Some NPs, such as Cu-NPs, may also release free metal ions from the surface of the particle. Biological uptake of NPs is not likely via ion transporters, but endocytosis is a possible uptake mechanism. The body distribution, metabolism, and excretion of nanometals is poorly understood and hampered by a lack of methods for measuring NPs in tissues. Although data sets are still limited, emerging studies on the acute toxicity of nanometals have so far shown that these materials can be lethal to fish in the mg-μgl(-1) range, depending on the type of material. Evidence suggests that some nanometals can be more acutely toxic to some fish than dissolved forms. For example, juvenile zebrafish have a 48-h LC(50) of about 0.71 and 1.78mgl(-1) for nano- and dissolved forms of Cu respectively. The acute toxicity of metal NPs is not always explained, or only partly explained, by the presence of free metal ions; suggesting that other novel mechanisms may be involved in bioavailability. Evidence suggests that nanometals can cause a range of sublethal effects in fish including respiratory toxicity, disturbances to trace elements in tissues, inhibition of Na(+)K(+)-ATPase, and oxidative stress. Organ pathologies from nanometals can be found in a range of organs including the gill, liver, intestine, and brain. These sublethal effects suggest some common features in the sublethal responses to nanometals compared to metal salts. Effects on early life stages of fish are also emerging, with reports of nanometals crossing the chorion (e.g., Ag-NPs), and suggestions that the nano-forms of some metals (Cu-NPs and ZnO NPs) may be more toxic to embryos or juveniles, than the equivalent metal salt. It remains possible that nanometals could interfere with, and/or stimulate stress responses in fish; but data has yet to be collected on this aspect. We conclude that nanometals do have adverse physiological effects on fish, and the hazard for some metal NPs will be different to the traditional dissolved forms of metals.     

Long-term dynamics of heavy metal concentrations in the food and liver of bank voles (<Emphasis Type="Italic">Myodes glareolus</Emphasis>) in the period of reduction of emissions from a copper smelter

Long-term changes in the contents of heavy metals (Cu, Zn, Cd, Рb) in the food and liver of bank voles (Myodes glareolus) inhabiting areas exposed to pollution from the Middle Ural Copper Smelter (MUCS) in the period of reduction of its emissions (1990–2015). The results show that 50-fold reduction of emissions has not resulted in an equivalent decrease in the dietary and body concentrations of metals: in the impact zone (1–2 km from the MUCS), Cu, Zn, and Pb concentrations remain unchanged, while Cd concentrations have increased twofold by the end of the observation period; in the background zone (20 km), Cu, Zn, and Cd concentrations remain unchanged, while Pb concentrations have decreased by a factor of 1.7–2.5; and no directed changes have been revealed in moderately polluted plots (4–6 km). The accumulation of heavy metals in the animal body depends primarily on the contents of these elements in food and on the system of elementspecific homeostatic barriers providing effective protection from the toxic effect of heavy metals.     

Effects of cadmium and zinc on larval growth and survival in the ground beetle,Pterostichus oblongopunctatus

Carabid beetles, like Pterostichus oblongopunctatus, living in metal contaminated areas may be exposed to elevated levels of metals within their diets. However, when compared to other second order consumers, they have one of the lowest observed levels of metals, indicating methods of detoxification to deal with such toxicants. In this study, we investigated if chronic, multigenerational exposure to metals leads to resistance to toxic metal concentrations, and if so, what are the costs associated with them. Adult organisms were collected from two sites, a polluted and a reference site near Olkusz, in southern Poland. These adults were immediately mated, and eggs were collected twice weekly to assess the effects in the larvae of the F(1) generation. Larvae were randomly exposed to one of four artificial mediums: control, 50 mg kg(-1) Cd, 500 mg kg(-1) Zn, and a combined treatment of 50 mg kg(-1) Cd and 500 mg kg(-1) Zn to investigate possible interactions. Individuals were sacrificed at 10, 30, and 40 days. Although metals were not accumulated in larvae (p>0.05), larvae fed the Cd or the Zn treatment grew significantly slower, and had the lowest survival rate (p<0.05) in respect to control. Out of metal treated animals, those on the combined treatment of CdZn grew the quickest and had the highest observed survival (p<0.05). Although previous studies have demonstrated changes in adult population parameters under chronic, multigeneration exposure to toxic metal concentrations, our study did not reveal any changes in the larval stage.     

Metal/metalloid contamination and isotopic composition of lead in edible mushrooms and forest soils originating from a smelting area

High metal contents in edible mushrooms growing in severely contaminated industrial areas pose an important toxicological risk. In the presented study, trace element (Pb, Cd, Zn, Cu, Ag, As, Se) contents were determined in caps and stipes of three different edible mushroom species (Boletus edulis Bull. Fr., Xerocomus badius Fr. Gilb., Xerocomus chrysenteron Bull. Quél.). Additionally, information about the chemical fractionation of metals in separate soil horizons and Pb isotopic data from soils and fruiting bodies allowed a more detailed insight on the uptake mechanisms of metals by the studied mushroom species. Total metal and metalloid concentrations in the organic soil horizons reached 36234 mg Pb kg(-1); 11.9 mg Cd kg(-1); 519 mg Zn kg(-1); 488 mg Cu kg(-1); 25.1 mg Ag kg(-1); 120 mg As kg(-1) and 5.88 Se mg kg(-1). In order to evaluate the accumulation capacity of the studied species, bioconcentration factors (BCF) were calculated for separate trace elements. For selected metals (Pb, Cd, Zn, Cu), a modified BCF calculation (using EDTA-extractable concentrations of metals in soil) was proposed. High contents of Pb (up to 165 mg kg(-1)) and Cd (up to 55 mg kg(-1)) exceeded all the regulatory limits in all the studied species. This was also the case for Se (up to 57 mg kg(-1)) in B. edulis. Intensive consumption of this species grown in such polluted areas can therefore pose toxicological risks for human health. A novel finding was that X. badius can act as an Ag accumulating species when grown at polluted sites due to the high concentrations of Ag (up to 190 mg kg(-1)) in caps. Pb isotopic data showed that Pb originating from the recent air pollution control residues is present mainly in the exchangeable/acid-extractable fraction of the organic horizons and is taken up by fruiting bodies; especially in the case of B. edulis, where fast Pb accumulation occurs. Due to the high species-dependent variations of metal contents, the studied mushrooms are not suitable as bioindicators of environmental pollution.     

Effect of elemental sulphur on solubility of soil heavy metals and their uptake by maize

A pot experiment was conducted to study the influence of elemental sulphur (S) on solubility of soil Pb, Zn and Cd and uptake by maize (Zea mays L.). Two rates of elemental sulphur (S) applied at 0 (S0) and 200 (S200) mmol kg(-1) soil with three rates of each heavy metal at Pb, 0 (Pb0), 200 (Pb200), 400 (Pb400) mg kg(-1) soil, Zn, 0 (Zn0), 100 (Zn100), 200 (Zn200) mg kg(-1) soil and Cd, 0 (Cd0), 50 (Cd50), 100 (Cd100) mg kg(-1) soil, respectively. The result showed that with S application at 200 mmol S kg(-1), soil pH decreased about 0.3 unit and the solubility of the Zn and Cd was significantly increased, but the solubility of Pb had no significant influence. The concentration of Pb, Zn and Cd in maize shoots and roots were increased with increasing rates of heavy metals. However, the concentration of Zn and Cd in shoots and roots were higher with application of S rather than without S but no significant difference was found for Pb. The highest concentration of Zn in the shoots was 2.3 times higher with application of S rather than without at the same rate of Zn, 200 mg kg(-1). Plant biomass was also significantly affected by the application of S and of heavy metals. With heavy metal addition, the shoot and root biomass were decreased with the rates of those of heavy metals increased either with or without application of S. However, the shoot biomass was significantly decreased with S application at the same rate of heavy metals except that with Zn addition. The removal of Cd and Pb by maize uptake and accumulation with application of S had no significant increase compared to that without, but the removal Zn by maize uptake from the soil increased by application of S, 90.9 microg plant(-1) contrast to 25.7 microg plant(-1) at Zn200 within a growth period of only 40 days.     

Occurrence and degradation of butyltins and wastewater marker compounds in sediments from Barcelona harbor, Spain

Contamination of Barcelona harbor sediments was assessed by the quantitative determination of butyltins (TBT, DBT and MBT) and surfactant intermediates, namely linear alkylbenzenes (LABs) and nonylphenols (NPs), as markers of urban and industrial wastewater contamination, respectively. Degradation indexes of TBT and LABs were calculated. Tributyltin predominated in the whole area over its degradation products, ranging from 98 to 4702 ng Sn/g. These elevated concentrations reveal a persistent historical contamination and a moderate degradation (BT(deg)). Moreover, the high LAB concentrations (1.2-53.1 microg/g) compared to the relatively low NP levels (3.8-72.0 ng/g) suggest a predominance of urban over industrial wastewater inputs, although a significant correlation (r(2) = 0.82, N = 12, P = 0.001) between LABs and NPs was found. Stormwater runoff and combined sewer overflows (CSO) were likely the most possible sources for both surfactant intermediates. The high degradation index values obtained for LABs could indicate an improvement in the wastewater management reducing its recent discharge into the Barcelona harbor area.     

Concentrations and loadings of polybrominated diphenyl ethers in dust from low-income households in California

California residents may experience the highest polybrominated diphenyl ether (PBDE) flame retardant exposures in the United States, the nation with the highest body burdens worldwide. It is hypothesized that Californians' high exposures are due to the state's strict furniture flammability standards. Ingestion of PBDE-contaminated dust, to which children may be particularly susceptible, is a dominant exposure pathway. Low-income populations may also face disparately high exposures due to the presence of older, deteriorated or poorly manufactured furniture treated with PBDEs. We collected up to two dust samples per home (54 samples total), several days apart, from low-income California households in the urban community of Oakland (n=13 homes) and the agricultural community of Salinas (n=15 homes). We measured BDE-47, BDE-99 and BDE-100, the major constituents of the penta-PBDE flame retardant formulation commonly used in furniture. All three PBDE congeners were detected in every sample with concentrations (loadings) ranging from 185 to 126,000ng/g (621-264,000ng/m(2)), 367-220,000ng/g (1550-457,000ng/m(2)), and 84-41,100ng/g (257-85,700ng/m(2)) for BDE-47, BDE-99 and BDE-100, respectively. Median concentrations (loadings) observed in Salinas homes for BDE-47, BDE-99 and BDE-100 were 3100ng/g (10,800ng/m(2)), 5480ng/g (19,500ng/m(2)), and 1060ng/g (3810ng/m(2)), respectively, and in Oakland homes 2780ng/g (10,700ng/m(2)), 4450ng/g (19,100ng/m(2)), and 1050ng/g (4000ng/m(2)), respectively. Maximum concentrations for BDE-47 and BDE-99 are the highest reported to date. Indoor concentrations and loadings did not significantly differ between communities; concentrations and loadings were strongly correlated between collections for all three congeners (Spearman rho=0.79-0.97, p<0.002). We estimated non-dietary ingestion of each congener for one child in each home (n=28 children) and found that estimated intake for BDE-47 and BDE-99 exceeded the U.S. Environmental Protection Agency's recommended chronic reference dose for three and five children, respectively. Children's estimated intake via dust ranged from 1.0 to 599ng/kg/day, 2.0-1065ng/kg/day and 0.5-196ng/kg/day for BDE-47, BDE-99 and BDE-100, respectively. In order to mitigate these exposures, future research must address the factors that contribute to PBDE exposures in low-income homes.     

Citric and Oxalic Acids Effect on Pb and Zn Uptake by Maize and Winter Wheat

A pot experiment was conducted to investigate the influence of citric and oxalic acids effect on Pb and Zn uptake by corn and winter wheat. The experiment was employed with citric acid (CA) applied at 3 rates (0, 1.5 and 3.0 mmol kg-1 soil), oxalic acid (OA) at 3 rates (0, 1.5 and 3.0mmol kg-1 soil) and citric acid combined with oxalic acid (1.5 mmol citric acid combined with 1.5 mmol oxalic acid kg-1). Two types of soil were chose in the experiment. One was collected from the agricultural soil near a battery-recycling factory in Anhui province, China (site A) and the other was collected from a Pb-Zn mine residues in Hunan province, China (site B). The results showed that soil pH varied with the different treatment of citric and oxalic acids. However, there were no differences in all the treatments. 3.0mmol CA kg-1 soil addition significantly increased the concentrations of the CaCI2-extractable Pb and Zn and other treatments have no significantly increased. The highest shoot concentrations of Pb and Zn     

上海市崇明岛公路两侧土壤重金属污染研究

采集了上海市崇明岛陈海、北沿公路两侧土壤和灰尘样品270余个,测定了样品的Pb、Cd、Cu、 Zn和Cr重金属含量。结果表明,陈海和北沿公路两侧土壤重金属Pb、Cd、Cu、 Zn和Cr的平均含量达到277、0279、258、918和776 mg/kg,土壤Cd污染较严重。采集的路面灰尘样品Pb、Cd、Cu、 Zn和Cr的平均含量达到512、049、489、209和970 mg/kg,超过土壤背景值2~4倍,是土壤重金属的主要二次污染源。公路防护林体系较差的北沿公路路侧土壤纵向剖面（垂直于公路走向）重金属含量随距路肩距离增加呈指数下降,土壤重金属重污染区在距路肩15 m范围内。防护林体系较完善的陈海公路距路肩15 m范围内土壤重金属污染较小,土壤重金属重污染区出现在距路肩20~50 m范围内。     

Modelling chronic exposure to contaminated soil: a toxicokinetic approach with the terrestrial snail Helix aspersa

To enlarge the possibilities of using organisms of the soil fauna to assess the bioaccumulative potential of chemicals, the kinetic of soil cadmium (Cd) transfer to the terrestrial gastropod Helix aspersa was investigated under laboratory conditions during a long-term experiment (6 months). During the exposure phase (3 months), juvenile snails were subjected to three different concentrations of Cd spiked in artificial ISO soil (ISO 0, 20 and 100 mg Cd kg(-1)) and to a field soil (ME4) industrially contaminated by 20 mg Cd kg (-1). For both soils, internal steady-state Cd concentrations were reached in the viscera of the snails, the main storage organ for Cd, after 2 weeks of exposure whatever the Cd concentration in soil. The equilibrium concentrations in the viscera were 0.7 (+/-0.1), 11.3 (+/-2.4), 73.3 (+/-4.8) and 6.3 (+/-1.3) mg Cd kg(-1) dry mass for ISO 0, ISO 20, ISO 100 and ME4, respectively. During the depuration phase (3 months), from 0 to 52% of the accumulated Cd in the viscera were removed by excretion or relocation in the foot. However, the snails were not able to depurate down to initial concentrations. Data were modelled by integrating a specific growth rate constant into one-compartment toxicokinetic models. This allowed the calculation of Cd uptake rates that can be used as indicators of metal bioavailability. Since this parameter was found to be lower for snails exposed to the field soil ME4, we concluded that lower Cd bioavailability in this field soil was responsible of the lower transfer to the snails compared to the ISO 20 soil, even though they were polluted to similar extents. Internal validation showed that the toxicokinetic models could be applied for predictive purposes, promising for the development of a bioaccumulation directive for terrestrial environment.     

Phthalate metabolites in urine from China, and implications for human exposures

Phthalates are esters of phthalic acid and are mainly used as plasticizers (added to plastics to increase their flexibility, transparency, durability, and longevity). Humans are exposed to phthalates through several routes. Urinary phthalate metabolites can be used as biomarkers of human exposures to phthalates. In this study, 14 phthalate metabolites were analyzed in 183 urine samples collected in 2010 from Shanghai, Guangzhou, and Qiqihaer, China. Phthalate metabolites were found in all urine samples and their total concentrations ranged from 18.6 to 3160 ng/mL (median: 331 ng/mL). Mono-n-butyl phthalate (mBP) and mono-2-isobutyl phthalate (miBP) were the major metabolites found in urine, and their respective median concentrations were 61.2 and 51.7 ng/mL; concentrations of miBP were higher than the concentrations reported for other countries, to date. Based on the urinary concentrations of phthalate metabolites, we estimated the daily intake rates in the Chinese population. The estimated daily intakes of dibutyl phthalate (DBP), diethyl phthalate (DEP), and di-(2-ethylhexyl) phthalate (DEHP) in China were 12.2, 3.8, and ~5 μg/kg bw/day, respectively. Thirty nine percent of the samples exceeded the tolerable daily intake of 10 μg/kg bw/day, proposed for DBP, by the European Food Safety Authority, but none of the estimated daily intake values exceeded the reference dose recommended by the U.S. Environmental Protection Agency.     

The toxicity of copper contaminated soil using a gnotobiotic Soil Multi-species Test System (SMS)

A gnotobiotic multi-species study was designed to consist of a food-web of soil-dwelling animals. The food-web was exposed to five concentrations copper (Cu) spiked soil for three exposure durations i.e. 28, 56 and 84 days. Based on multivariate analysis the food-web was significantly affected by Cu exposure at and above 300 mg Cu kg(-1) soil (lowest tested concentration). The number of animals present in the 2500 mg Cu kg(-1) (highest tested concentration) was at all sampling occasions below the starting point level. Based on analysis of the individual species the lowest 10% effect concentration (EC10) observed was 50 mg Cu kg(-1) soil, for Enchytraeus crypticus. Using the EC10 for the individual species the HC5 (Hazard Concentration at the 5% level) was estimated to be between 25 and 36 mg Cu kg(-1) soil, depending on the exposure duration. A similar experiment but using a reduced design was performed employing soil contaminated with Cu in the field more than 80 years ago. The trend in the field-contaminated soil was similar to that observed for the spiked soil.