Response of Artemisia herba alba to hexavalent chromium pollution under arid and semi-arid conditions.

The aim of this work is to study and evaluate in situ, the Artemisia herba alba responses to hexavalent chromium stress in the arid and semi-arid steppe Algerian soil. This metallic pollutant was selected to its high toxicity and to its great release from several industrial and agricultural activities emissions in the area of the study region. Artemisia herba alba is a medicinal plant but also a forage species widely used in pasture. It has dominated then adapted to the arid and semi-arid climate of Algerian steppe region, due principally to their morphological and physiological characteristics. To establish this work, A. herba alba species were selected in the Algerian steppe region, and their soils were treated weekly with K2CrO4 solution for about three months. Chromium concentrations were determined in the soil and in the different plants' parts to verify its absorption and translocation with and without pollution simulation. This study demonstrated that A. herba alba has a strong resistance to high concentrations of hexavalent chromium. An increase in chromium concentrations at the different parts of plant's has been noticed, without affecting its growth and vegetative development. The results of physiological and elementary analysis indicated that chromium was absorbed by A. herba alba. Cr (VI) accumulation in plant increase when they are increasing in soil. Translocation factor results indicated that Cr (VI) was mainly accumulated by A. herba alba roots. Particle size analysis showed that the soil is poor in organic substances and mostly sandy loams with 79.77% of sand and 9.76% of clay minerals.

Effects of dissolved organic matter derived from forest leaf litter on biodegradation of phenanthrene in aqueous phase.

Dissolved organic matter (DOM) released from forest leaf litter is potentially effective for the degradation of polycyclic aromatic hydrocarbons (PAHs), yet the inherent mechanism remains insufficiently elucidated. In this study, we investigated the effects of DOM derived from Pinus elliottii and Schima superba leaf litter on the degradation of phenanthrene by the phenanthrene degrading bacterium Sphingobium sp. Phe-1. DOM from different origins and at a large range of concentrations enhanced the degradation rate of phenanthrene. DOM derived from P. elliottii leaf litter decomposed for 12 months used at a concentration of 100mg/L yielded the highest degradation rate (16.9% in 36h) and shortened the degradation time from 48h to 24h. Changes in the composition of DOM during degradation as measured by EEMs-FRI showed that proteins and tyrosine in the DOM supplied readily available nutrients that stimulated biological activity of Phe-1, increasing its growth rate and catechol 2,3-dioxygenase activity. Simultaneously, fulvic acid and humic acid in the DOM enhanced phenanthrene bioavailability by increasing the solubility and mass transfer of phenanthrene, enhancing the uptake kinetics of Phe-1, and increasing the bacteria's direct access to DOM-associated phenanthrene. Humic acid was co-metabolized by Phe-1, resulting in further stimulation of phenanthrene degradation.

Physico-chemical characteristics of topsoil for contrasted forest, agricultural, urban and industrial land uses in France.

Soil quality is related to soil characteristics such as fertility and contamination. The aim of this study is to assess the effect of land use on these soil characteristics and to confirm the following anthropisation gradient: (i) forest, (ii) grassland, (iii) cultivated, (iv) orchard and vineyard, (v) urban vegetable garden, and (vi) SUITMA (urban, industrial, traffic, mining and military areas). A database comprising the characteristics of 2451 soils has been constituted. In order to compare the topsoils from six contrasting land uses, a principal components analysis (PCA) was performed on nine geochemical variables (C, N, pH, POlsen, total Cd, Cu, Ni, Pb, Zn). The first axis of the PCA is interpreted as a global increase of topsoil metallic elements along the anthropisation gradient. Axis 2 reflects the variability of fertility levels. Human activity increases the pressure on soils along the proposed gradient according to six different distribution patterns. This better knowledge of topsoil quality and its dependence on current land use should therefore help to manage and preserve the soil mantle.

Factors influencing the Zn and Mn extraction from pyrometallurgical sludge in the steel manufacturing industry.

In this laboratory study, a process has been developed for selectively leaching zinc and manganese from pyrometallurgical sludge produced in the steel manufacturing industry. In the first part, the yield of Zn extraction was studied using four factors and four levels of the Box-Behnken response surface design. The optimum conditions for the step of Zn leaching were determined to be a sulfuric acid concentration of 0.25 mol/L, a pulp density of 10%, an extraction temperature of 20 °C, and three stages of leaching. Under such conditions, 75% of the Zn should be leached. For Mn leaching, the optimum conditions were determined to be a sulfuric acid concentration of 0.25 mol/L, a Na2S2O5/Mn stoichiometry of 1, a leaching time of 120 min and two leaching steps. In this case, 100% of the Mn should be leached.

PAH phytoremediation: rhizodegradation or rhizoattenuation?

Dealing with soil contaminated with persistent organic pollutants (POP) is an increasing concern amplified by both regulatory constraints and the dramatic impact of human activities on the soil resource. The most used management options are treatments which totally eradicate the toxic compounds targeted. When possible, environmental-friendly processes should be used, and recent years have seen the emergence of green technologies using biological energies involving microorganisms (bioremediation) and plants (phytoremediation). Research has focused on phytoremediation and many have presented this technology as the process ideally combining efficiency, low cost and environmental acceptance. However, the applicability of phytoremediation on soils contaminated by bio-recalcitrant organic compounds, such as polycyclic aromatic hydrocarbons (PAH), has not yet proved as successful as expected. We propose here a review and discussion of the overall question of PAH status in soil and their potential for treatment. The limits and applicability of bioremediation technologies are discussed, and the specific beneficial effect of plants is objectively evaluated with a special interest to processes which lead to rhizoattenuation. Given the PAH high affinity to soil organic matter, availability is the main limitation to phytoremediation. In this context, bioavailability quantification remains an issue as well as the characterization of the recalcitrant fraction.

A new process for nickel ammonium disulfate production from ash of the hyperaccumulating plant Alyssum murale.

The extraction of nickel (Ni) from ultramafic soils by phytomining can be achieved using Alyssum murale cultures. This study presents a new process for the valorization of Ni accumulated by this plant through the production of a Ni ammonium disulfate salt (Ni(NH(4))(2)(SO(4))(2).6H(2)O). The process comprises an initial leaching of the ashes of A. murale with a sulphuric acid solution (1.9 M H(2)SO(4), T=95 °C, t=240 min, TS=150 g ash L(-1)), producing a leachate rich in Ni (10.2 g Ni L(-1); 96% Ni solubilisation), Mg, P, K, Fe, Ca and Al. The pH of the acid leachate is increased to 5.0 with NaOH (5M), followed by an evaporation step which produced a purified solution rich in Ni (21.3 g NiL(-1)) and an iron hydroxide precipitate. The cold crystallization (T=2 °C, t=6h) of this solution by the stoichiometric addition (× 1.2) of ammonium sulfate generates a Ni ammonium disulfate salt, containing 13.2% Ni, that is potentially valuable to industry.

In situ assessment of phytotechnologies for multicontaminated soil management.

Due to human activities, large volumes of soils are contaminated with organic pollutants such as polycyclic aromatic hydrocarbons, and very often by metallic pollutants as well. Multipolluted soils are therefore a key concern for remediation. This work presents a long-term evaluation of the fate and environmental impact of the organic and metallic contaminants of an industrially polluted soil under natural and plant-assisted conditions. A field trial was followed for four years according to six treatments in four replicates: unplanted, planted with alfalfa with or without mycorrhizal inoculation, planted with Noccaea caerulescens, naturally colonized by indigenous plants, and thermally treated soil planted with alfalfa. Leaching water volumes and composition, PAH concentrations in soil and solutions, soil fauna and microbial diversity, soil and solution toxicity using standardized bioassays, plant biomass, mycorrhizal colonization, were monitored. Results showed that plant cover alone did not affect total contaminant concentrations in soil. However, it was most efficient in improving the contamination impact on the environment and in increasing the biological diversity. Leaching water quality remained an issue because of its high toxicity shown by micro-algae testing. In this matter, prior treatment of the soil by thermal desorption proved to be the only effective treatment.

The influence of thermal desorption on genotoxicity of multipolluted soil.

A multipolluted soil sampled from a former coking plant in Lorraine (France) was evaluated for its genotoxic effects on coelomocytes of the Eisenia fetida earthworm using the comet assay. The biological efficiency of thermal desorption of the contaminated soil was also investigated. The untreated polluted soil was shown to be genotoxic to earthworms. Although thermal desorption reduced the concentration of PAHs by 94% (Sigma(16 PAHs)=1846 and 101 mg/kg before and after thermal desorption, respectively), the treatment did not eliminate the genotoxicity of soil pollutants to earthworms but increased it. The concentration of non-volatile metals did not change after thermal desorption. Among metals found in the treated soil, cadmium, chromium and nickel could explain the genotoxicity of the contaminated soil after thermal desorption. The treatment could increase the bioavailability and genotoxicity of heavy metals, through a modification of the soil's organic matter, the speciation of heavy metals and their binding to organic matter. This study underlines the importance of measuring biological effects, in order to evaluate the risk associated with formerly contaminated soils and the efficiency of remediation.

Bioavailability of chemical pollutants in contaminated soils and pitfalls of chemical analyses in hazard assessment.

Decision-making for remediation of industrial wastelands are still based on the concentrations of pollutants of concern measured in soils. In this work, two soils polluted by polycyclic aromatic hydrocarbons (PAHs) and metals were investigated for their toxicity on earthworms (Eisenia fetida), collembolae (Folsomia candida), and higher plants (Brassica chinensis, Lactucca sativa and Avena sativa) in order to study the relationships between chemical contamination and biological effects. Although the level of contamination by PAHs was elevated and commensurate in the two soils, their toxicity profile was quite different. Soil A affected survival and reproduction of invertebrates and growth of higher plants. Surprisingly, soil B, heavily contaminated by metals in addition to PAHs, was devoid of toxicity. Our results indicate that toxicity cannot simply be extrapolated from pollutant concentrations in a complex matrix in which bioavailability of pollutants may be reduced by ageing. Moreover, the use of toxicity data obtained from spiked soils characterized by readily bioavailable pollutants can also be called into question for such extrapolations. Predicting biological effects therefore requires biological tools to avoid any erroneous conclusions that can be drawn from sole extrapolation of analytical results.

Soil microbial diversity as affected by the rhizosphere of the hyperaccumulator Thlaspi caerulescens under natural conditions.

It is hypothesized that metal hyperaccumulator plants have specific rhizosphere conditions, potentially modifying the bioavailability of soil metals. This article aims to further the knowledge about the rhizosphere of the hyperaccumulator Thlaspi caerulescens, focusing on its microflora isolated from metalliferous soils collected in situ where the plants grow naturally. We characterized the cultivable microbial communities isolated from the rhizosphere of one population of this Ni hyperaccumulator species grown on a serpentine soil. The rhizosphere soil harbored a wide variety of microorganisms, predominantly bacteria, confirming the stimulatory effect of the T. caerulescens rhizosphere on microbial growth and proliferation. We tested the hypothesis that the rhizosphere of T. caerulescens influences (1) the metabolic diversity of the bacterial community and (2) the bacterial resistance to metals. The principal component analysis of the Biolog plate's data confirmed a structural effect of the rhizosphere of T. caerulescens on bacterial communities. The percentage of Ni-resistant bacteria was higher in the rhizosphere than in the bulk soil, suggesting a direct effect of the rhizosphere on Ni tolerance, reflecting a greater bacterial tolerance to Ni in the rhizosphere.

Retention and phytoavailability of radioniobium in soils.

Radioniobium is present in long-lived nuclear waste as a result of the activation of zirconium pellets associated with the nuclear fuel. The behaviour of niobium (Nb) in the environment and especially its fate in the soil-plant system has not been thoroughly investigated so far. In safety assessment of French long-lived nuclear waste disposal, data concerning the mobility and the bioavailability of Nb in soils are needed as well as general trends of its fate in the specific environment around the site of French underground research laboratory. Therefore, we investigated the mobility of 95Nb in three different soils typical of the area of north-eastern France and its uptake by two plants, rye grass (Lolium perenne L.) and winter wheat (Triticum aestivum L.). Soil:solution distribution of 95Nb was observed in 1:10 batch experiments with deionized water for a 3-day period. Results showed that K(d) values were high (in the order of 10(3) L kg(-1)) and were still significantly increasing after 3 days. A mathematical model, fitted to describe the decrease of the radioactivity after 3 days, is proposed to calculate sorption ratios--SR--(rather than K(d) values as equilibrium was not reached) over longer periods. Soil-to-plant concentration ratios (CR) were measured in shoots and roots of the two plants after cultivation on two soils spiked with (95)Nb (406kBqkg(-1)). Soil-to-root dry weight CR were high (0.30-1.52) and could probably be due to efficient uptake into the roots. However, no transfer of Nb to plant shoots was detected in any of the soils. Nb is thus a rather immobile element in soils and its transfer to plants seems limited to underground parts. It would therefore tend to accumulate in surface horizons of soils in case of long-term continuous surface release.

Assessment of plant uptake of radioactive nickel from soils.

As a result of isotopic dilution, the availability to plants of radioisotopes introduced into the soil solution should be directly related to the size of the isotopically exchangeable pool (E(t))-value). This work was undertaken to test this hypothesis for the radionuclide 63Ni. The demonstration was based on pot experiments conducted with seven soils representing a large range of Ni content (from 9.9 mg kg(-1) to 862.6 mg kg(-1)) which were mixed with a 63NiCl2 solution (100 kBq kg(-1)). Three plant species varying in Ni uptake, Triticum aestivum (wheat), Trifolium pratense (clover), and the Ni-hyperaccumulator Alyssum murale, were grown for 90 d, and their total Ni and 63Ni content determined at harvest. In parallel, the isotopically exchangeable kinetics method (IEK) was run on each soil sample to measure the E(t)-value. Results showed that plant uptake of radioactive nickel was negatively correlated with the E(t)-value with wheat and clover as a result of the dilution of 63Ni added in the isotopically exchangeable pool of soil Ni (alpha=5%); correlation was positive with the A. murale (alpha=10%). Hence, this provides a new approach for the assessment of soil-to-plant transfer of 63Ni at larger scale avoiding the carrying out of time consuming experiments.

Identification and function of ryanodine receptor subtype 3 in non-pregnant mouse myometrial cells.

Subtype 3 of the ryanodine receptor (RYR3) is a ubiquitous Ca2+ release channel which is predominantly expressed in smooth muscle tissues and certain regions of the brain. We show by reverse transcription-polymerase chain reaction (RT-PCR) that non-pregnant mouse myometrial cells expressed only RYR3 and therefore could be a good model for studying the role of endogenous RYR3. Expression of RYR3 was confirmed by Western blotting and immunostaining. Confocal Ca2+ measurements revealed that in 1.7 mM extracellular Ca2+, neither caffeine nor photolysis of caged Ca2+ were able to trigger any Ca2+ responses, whereas in the same cells oxytocin activated propagated Ca2+ waves. However, under conditions of increased sarcoplasmic reticulum (SR) Ca2+ loading, brought about by superfusing myometrial cells in 10 mM extracellular Ca2+, all the myometrial cells responded to caffeine and photolysis of caged Ca2+, indicating that it was possible to activate RYR3. The caffeine-induced Ca2+ responses were inhibited by intracellular application of an anti-RYR3-specific antibody. Immunodetection of RYR3 with the same antibody revealed a rather homogeneous distribution of fluorescence in confocal cell sections. In agreement with these observations, spontaneous or triggered Ca2+ sparks were not detected. In conclusion, our results suggest that under conditions of increased SR Ca2+ loading, endogenous RYR3 may contribute to the Ca2+ responses of myometrial cells.

Measurement of in situ phytoextraction of zinc by spontaneous metallophytes growing on a former smelter site.

This work was undertaken to measure the in situ phytoextraction of zinc using a former zinc-smelter site where metallophyte plants have been growing for 30 years. The site exhibited a gradient in the total metal concentration in the upper horizon (from 3230 to 8530 mg Zn kg(-1)). Soils were sampled from four different sectors (I-IV), and plant shoots were harvested, identified, their biomass weighed and analysed for zinc. The results showed that three plant species were dominant on the site, including Arabidopsis halleri (cress), Armeria maritima (seathrift), and Arrhenatherum elatius (fromental). A. maritima was the predominant species according to the biomass production on the most polluted sector 1. As the concentration of metals in soils decreased. A. maritima disappeared and A. halleri increased. The biomass of A. elatius was the highest on the less polluted soils. Concentrations in zinc in the aerial parts of plants varied from 73 (sector IV) to 6269 mg kg(-1) DM (sector 1). The concentration of Zn in A. halleri decreased with the decrease in concentration of zinc in soil. Phytoextraction was calculated from the biomass and its concentration of metal. It was at a maximum in sector III with a high contribution of A. halleri and A. elatius and reached 10 kg Zn ha(-1), a promising amount for phytoextraction considering the absence of any agricultural practices. In sector 1, phytoextraction was four times lower despite a 2.6 times higher concentration of Zn in the upper horizon. In conclusion, phytoextraction was strongly dependent on the concentration of the available metal in soils which may limit the growth of plants, and favour tolerant but low biomass plant species such as A. maritima.

Calcium signalling through nucleotide receptor P2X1 in rat portal vein myocytes.

1. ATP-mediated Ca2+ signalling was studied in freshly isolated rat portal vein myocytes by means of a laser confocal microscope and the patch-clamp technique. 2. In vascular myocytes held at -60 mV, ATP induced a large inward current that was supported mainly by activation of P2X1 receptors, although other P2X receptor subtypes (P2X3, P2X4 and P2X5) were revealed by reverse transcription-polymerase chain reaction. 3. Confocal Ca2+ measurements revealed that ATP-mediated Ca2+ responses started at initiation sites where spontaneous or triggered Ca2+ sparks were not detected, whereas membrane depolarizations triggered Ca2+ waves by repetitive activation of Ca2+ sparks from a single initiation site. 4. ATP-mediated Ca2+ responses depended on Ca2+ influx through non-selective cation channels that activated, in turn, Ca2+ release from the intracellular store via ryanodine receptors (RYRs). Using specific antibodies directed against the RYR subtypes, we show that ATP-mediated Ca2+ release requires, at least, RYR2, but not RYR3. 5. Our results suggest that, in vascular myocytes, Ca2+ influx through P2X1 receptors may trigger Ca2+-induced Ca2+ release at intracellular sites where RYRs are not clustered.

Effect of sludge-amendment or nutrient addition on the biodegradation of the herbicide isoproturon in soil.

Adding sludge to agricultural soil results in added organic matter, nutrients and metallic and/or organic pollutants. These components may modify the behaviour of pesticides in the soil. We monitored possible changes in the degradation of the herbicide isoproturon (production of CO2 and degradation products) in soil amended with sludge, heavy metals or nitrogen and phosphorus. The treated and control soils were incubated under controlled conditions for 60 days. The nitrogen and phosphorus had the greatest effect on isoproturon degradation, independent of the presence of pollutants. Mineralisation of the herbicide to CO2 was slow and seemed to be linked to a fast degradation and to the accumulation of a complex degradation product that was neither catabolized nor adsorbed, 4,4'-diisopropylazobenzene. This degradation pathway also produced smaller amounts of non-extractable residues. Sewage sludge had no significant effect on isoproturon degradation, despite a large increase of organic matter mineralisation (factor 2).

Ca(2+) signals mediated by Ins(1,4,5)P(3)-gated channels in rat ureteric myocytes.

Localized Ca(2+)-release signals (puffs) and propagated Ca(2+) waves were characterized in rat ureteric myocytes by confocal microscopy. Ca(2+) puffs were evoked by photorelease of low concentrations of Ins(1,4,5)P(3) from a caged precursor and by low concentrations of acetylcholine; they were also observed spontaneously in Ca(2+)-overloaded myocytes. Ca(2+) puffs showed some variability in amplitude, time course and spatial spread, suggesting that Ins(1,4,5)P(3)-gated channels exist in clusters containing variable numbers of channels and that within these clusters a variable number of channels can be recruited. Immunodetection of Ins(1,4,5)P(3) receptors revealed the existence of several spots of fluorescence in the confocal cell sections, supporting the existence of clusters of Ins(1,4,5)P(3) receptors. Strong Ins(1,4,5)P(3) photorelease and high concentrations of acetylcholine induced Ca(2+) waves that originated from an initiation site and propagated in the whole cell by spatial recruitment of neighbouring Ca(2+)-release sites. Both Ca(2+) puffs and Ca(2+) waves were blocked selectively by intracellular applications of heparin and an anti-Ins(1,4,5)P(3)-receptor antibody, but were unaffected by ryanodine and intracellular application of an anti-ryanodine receptor antibody. mRNAs encoding for the three subtypes of Ins(1,4,5)P(3) receptor and subtype 3 of ryanodine receptor were detected in these myocytes, and the maximal binding capacity of [(3)H]Ins(1,4,5)P(3) was 10- to 12-fold higher than that of [(3)H]ryanodine. These results suggest that Ins(1,4,5)P(3)-gated channels mediate a continuum of Ca(2+) signalling in smooth-muscle cells expressing a high level of Ins(1,4,5)P(3) receptors and no subtypes 1 and 2 of ryanodine receptors.

Beta-3 adrenergic stimulation of L-type Ca(2+) channels in rat portal vein myocytes.

1. The effects of beta(3)-adrenergic stimulation were studied on the L-type Ca(2+) channel in single myocytes from rat portal vein using the whole-cell mode of the patch-clamp technique. 2. Reverse transcription-polymerase chain reaction showed that beta(1)-, beta(2)- and beta(3)-adrenoceptor subtypes were expressed in rat portal vein myocytes. Application of both propranolol (a non-selective beta(1)- and beta(2)-adrenoceptor antagonist) and SR59230A (a beta(3)-adrenoceptor antagonist) were needed to inhibit the isoprenaline-induced increase in L-type Ca(2+) channel current. 3. L-type Ca(2+) channels were stimulated by CGP12177A (a beta(3)-adrenoceptor agonist with potent beta(1)- and beta(2)-adrenoceptor antagonist property) in a manner similar to that of isoprenaline. The CGP12177A-induced stimulation of Ca(2+) channel current was blocked by SR59230A, cyclic AMP-dependent protein kinase inhibitors, H-89 and Rp 8-Br-cyclic AMPs, but was unaffected by protein kinase C inhibitors, GF109203X and 19-31 peptide. This stimulation was mimicked by forskolin and 8-Br-cyclic AMP. In the presence of okadaic acid (a phosphatase inhibitor), the beta(3)-adrenoceptor-induced stimulation was maintained after withdrawal of the agonist. 4. The beta(3)-adrenoceptor stimulation of L-type Ca(2+) channels was blocked by a pretreatment with cholera toxin and by the intracellular application of an anti-Galpha(s) antibody. This stimulation was unaffected by intracellular infusion of an anti-Gbeta(com) antibody and a betaARK(1) peptide. 5. These results show that activation of beta(3)-adrenoceptors stimulates L-type Ca(2+) channels in vascular myocytes through a Galpha(s)-induced stimulation of the cyclic AMP/protein kinase A pathway and the subsequent phosphorylation of the channels.

Requirement of ryanodine receptor subtypes 1 and 2 for Ca(2+)-induced Ca(2+) release in vascular myocytes.

While the roles of subtypes 1 and 2 of the ryanodine receptors (RYRs) have been studied in cellular systems expressing specifically one or the other of these subtypes (i.e. skeletal and cardiac muscle), the function of these receptors has not been evaluated in smooth muscles. We have previously reported RYR-mediated elementary (Ca(2+) sparks) and global Ca(2+) responses in rat portal vein myocytes. Here, we investigated the respective roles of all three RYR subtypes expressed in these cells as revealed by reverse transcriptase-polymerase chain reaction. Antisense oligonucleotides targeting each one of the three RYR subtypes were shown to specifically inhibit the expression of the corresponding mRNA and protein without affecting the other RYR subtypes. Confocal Ca(2+) measurements revealed that depolarization-induced Ca(2+) sparks and global Ca(2+) responses were blocked when either RYR1 or RYR2 expression was suppressed. Caffeine-induced Ca(2+) responses were partly inhibited by the same antisense oligonucleotides. Neither the corresponding scrambled oligonucleotides nor the antisense oligonucleotides targeting RYR3 affected depolarization- or caffeine-induced Ca(2+) responses. Our results show that, in vascular myocytes, the two RYR1 and RYR2 subtypes are required for Ca(2+) release during Ca(2+) sparks and global Ca(2+) responses, evoked by activation of voltage-gated Ca(2+) channels.

Assessment of metal content and toxicity of leachates from teapots.

Metallic teapots traditionally used in Morocco were investigated for release of toxic metals from the teapots and their toxicity, as determined by MetPAD, a bacterial toxicity test that is specific for heavy metal toxicity. Our data show that some teapots were non-toxic while a few others were highly toxic, as shown by MetPAD. Tea addition reduced somewhat heavy metal toxicity due possibly to the complexing ability of tea. Chemical analysis of teapot leachates showed that some contained zinc and copper. Teapot No. 5, which showed the highest toxicity, also displayed the highest Zn concentration (7.39 mg/l), confirming the toxicity data. Based on estimates of tea consumption in Morocco, we showed that the extra daily burden of Zn ranged from 1.75 to 4.2 mg/day, assuming the maximum zinc concentration of 7.4 mg/l, as found in our study. This represents 3.5-8% of the LOAEL for zinc of 50 mg/day and would not be important as compared to other sources zinc intake.