Assessment of the mutagenic potential of arecoline in gpt delta transgenic mice

Until recently, knowledge about the genotoxicity of roxarsone in vitro or in vivo was limited. This study assessed the genotoxicity of roxarsone in an in vitro system. Roxarsone was tested for potential genotoxicity on V79 cells by a Comet assay and a micronucleus (MN) test, exposing the cells to roxarsone (1-500 μM) and to sodium arsenite (NaAsO?, 20 μM) solutions for 3-48 h. Roxarsone was found to be cytotoxic when assessed with a commercial cell counting kit (CCK-8) used to evaluate cell viability, and moderately genotoxic in the Comet assay and micronucleus test used to assess DNA damage. The Comet metrics (percentages TDNA, TL, TM) increased significantly in a time- and concentration-dependent manner in roxarsone-treated samples compared with PBS controls (P<0.05), while the data from samples treated with 20 μM NaAsO? were comparable to those from 500 μM roxarsone-treated samples. The MN frequency of V79 cells treated with roxarsone was higher than that in the negative control but lower than the frequency in cells treated with 20 μM NaAsO?. A dose- and time-dependent response in MN induction was observed at 10, 50, 100 and 500 μM doses of roxarsone after 12-48 h exposure time. The DNA damage in V79 cells treated with 500 μM roxarsone was similar to cells exposed to 20 μM NaAsO?. The uptake of cells was correlated with the DNA damage caused by roxarsone. This investigation depicts the genotoxic potentials of roxarsone to V79 cells, which could lead to further advanced studies on the genotoxicity of roxarsone.     

Micronucleus frequencies in workers exposed to lead, zinc, and cadmium

Micronuclei (MN) in blood lymphocytes were determined in 31 male workers occupationally exposed to lead (Pb), zinc (Zn), and cadmium (Cd) and 20 control workers matched for age and smoking habits. Exposed workers have higher MN mean values than control workers (p<0.01). In exposed workers, blood Pb concentrations were also significantly higher than in control workers (p<0.001), but the mean concentrations of Zn and Cd in the blood were not statistically significant compared to the controls (p>0.05). These results suggest that lead may be genotoxic and the human lymphocyte micronucleus test can be used to assess genotoxic effects that result from occupational exposures.     

Accumulation, detoxification, and genotoxicity of heavy metals in Indian mustard (Brassica juncea L.)

Plants of Indian mustard (Brassica juncea L.) were exposed to different concentrations (15, 30, 60, 120 microM) of (Cd, Cr, Cu, Pb) for 28 and 56 d for accumulation and detoxification studies. Metal accumulation in roots and shoots were analyzed and it was observed that roots accumulated a significant amount of Cd (1980 microg g(-1) dry weight), Cr (1540 microg g(-1) dry weight), Cu (1995 microg g(-1) dry weight), and Pb (2040 microg g(-1) dry weight) after 56 d of exposure, though in shoot this was 1110, 618, 795, and 409 microg g(-1) dry weight of Cd, Cr, Cu, and Pb, respectively. In order to assess detoxification mechanisms, non-protein thiols (NP-SH), glutathione (GSH) and phytochelatins (PCs) were analyzed in plants. An increase in the quantity of NP-SH (9.55), GSH (8.30), and PCs (1.25) micromol g(-1) FW were found at 15 microM of Cd, however, a gradual decline in quantity was observed from 15 microM of Cd onwards, after 56 d of exposure. For genotoxicity in plants, cytogenetic end-points such as mitotic index (MI), micronucleus formation (MN), mitotic aberrations (MA) and chromosome aberrations (CA) were examined in root meristem cells of B. juncea. Exposure of Cd revealed a significant (P < 0.05) inhibition of MI, induction of MA, CA, and MN in the root tips for 24 h. However, cells examined at 24 h post-exposure showed concentration-wise recovery in all the endpoints. The data revealed that Indian mustard could be used as a potential accumulator of Cd, Cr, Cu, and Pb due to a good tolerance mechanisms provided by combined/concerted action of NP-SH, GSH, and PCs. Also, exposure of Cd can cause genotoxic effects in B. juncea L. through chromosomal mutations, MA, and MN formation.     

Involvement of the synapse-specific zinc transporter ZnT3 in cadmium-induced hippocampal neurotoxicity

The present study examined the involvement of zinc (Zn)-transporters (ZnT3) in cadmium (Cd)-induced alterations of Zn homeostasis in rat hippocampal neurons. We treated primary rat hippocampal neurons for 24 or 48 hr with various concentrations of CdCl₂ (0, 0.5, 5, 10, 25, or 50 μM) and/or ZnCl ₂ (0, 10, 30, 50, 70, or 90 μM), using normal neuronal medium as control. By The CellTiter 96 ^® Aqueous One Solution Cell Proliferation Assay (MTS; Promega, Madison, WI) assay and immunohistochemistry for cell death markers, 10 and 25 μM of Cd were found to be noncytotoxic doses, and both 30 and 90 μM of Zn as the best concentrations for cell proliferation. We tested these selected doses. Cd, at concentrations of 10 or 25 μM (and depending on the absence or presence of Zn), decreased the percentage of surviving cells. Cd-induced neuronal death was either apoptotic or necrotic depending on dose, as indicated by 7-AAD and/or annexin V labeling. At the molecular level, Cd exposure induced a decrease in hippocampal brain-derived neurotrophic factor-tropomyosin receptor kinase B (BDNF-TrkB) and Erk1/2 signaling, a significant downregulation of the expression of learning- and memory-related receptors and synaptic proteins such as the NMDAR NR2A subunit and PSD-95, as well as the expression of the synapse-specific vesicular Zn transporter ZnT3 in cultured hippocampal neurons. Zn supplementation, especially at the 30 μM concentration, led to partial or total protection against Cd neurotoxicity both with respect to the number of apoptotic cells and the expression of several genes. Interestingly, after knockdown of ZnT3 by small interfering RNA transfection, we did not find the restoration of the expression of this gene following Zn supplementation at 30 μM concentration. These data indicate the involvement of ZnT3 in the mechanism of Cd-induced hippocampal neurotoxicity.     

Chromosomal changes: induction, detection methods and applicability in human biomonitoring

The objective of this state of the art paper is to review the mechanisms of induction, the fate, the methodology, the sensitivity/specificity and predictivity of two major cytogenetic endpoints applied for genotoxicity studies and biomonitoring purposes: chromosome aberrations and micronuclei. Chromosomal aberrations (CAs) are changes in normal chromosome structure or number that can occur spontaneously or as a result of chemical/radiation treatment. Structural CAs in peripheral blood lymphocytes (PBLs), as assessed by the chromosome aberration (CA) assay, have been used for over 30 years in occupational and environmental settings as a biomarker of early effects of genotoxic carcinogens. A high frequency of structural CAs in lymphocytes (reporter tissue) is predictive of increased cancer risk, irrespective of the cause of the initial CA increase. Micronuclei (MN) are small, extranuclear bodies that arise in dividing cells from acentric chromosome/chromatid fragments or whole chromosomes/chromatids that lag behind in anaphase and are not included in the daughter nuclei in telophase. The cytokinesis-block micronucleus (CBMN) assay is the most extensively used method for measuring MN in human lymphocytes, and can be considered as a "cytome" assay covering cell proliferation, cell death and chromosomal changes. The key advantages of the CBMN assay lie in its ability to detect both clastogenic and aneugenic events and to identify cells which divided once in culture. Evaluation of the mechanistic origin of individual MN by centromere and kinetochore identification contributes to the high sensitivity of the method. A number of findings support the hypothesis of a predictive association between the frequency of MN in cytokinesis-blocked lymphocytes and cancer development. Recent advances in fluorescence in situ hybridization (FISH) and microarray technologies are modifying the nature of cytogenetics, allowing chromosome and gene identification on metaphase as well as in interphase. Automated scoring by flow cytometry and/or image analysis will enhance their applicability.     

Genotoxic effects in M5 cells and Chinese hamster V79 cells after exposure to 7Li-beam (LET=60 keV/microm) and correlation of their survival dynamics to nuclear damages and cell death

Chinese hamster V79 cell and a cell strain M5, derived from V79 cells and reported to be relatively resistant to gamma-ray, hydrogen peroxide, and N-methyl-N-nitro-N-nitrosoguanidine (MNNG; a potent human carcinogen), were exposed to high LET (7)Li-beam (LET=60 keV/microm) at approximately 90% confluent state in the dose range of 0-1 Gy. Effects of (7)Li-beam exposure on cell survival, micronuclei induction (MN), chromosomal aberrations (CA) and apoptosis were compared in both the cell lines. A dose-dependent decline in survival for both the cell lines was noted, relatively less in M5 cells (mostly p<0.01) indicating greater radio-resistance in this strain. The MN, CA and apoptosis increased in a dose-dependent manner in both V79 and M5 cells. Significant differences in various other parameters between these two cell lines were also noted. The relative intensity of DNA ladder, which is a useful marker for the determination of the extent of apoptosis induction, was much higher in V79 cells. A good correlation between the reduction of the surviving fractions and the increase in frequencies of MN or CA or apoptosis was noted for both the cell lines.     

Genotoxic effects of occupational exposure to lead and cadmium

This study was designed to assess genotoxic damage in somatic cells of workers in a Polish battery plant after high-level occupational exposure to lead (Pb) and cadmium (Cd), by use of the following techniques: the micronucleus (MN) assay, combined with in situ fluorescence hybridization (FISH) with pan-centromeric probes, analysis of sister chromatid exchanges (SCEs), and the comet assay. Blood samples from 44 workers exposed to lead, 22 exposed to cadmium, and 52 unexposed persons were used for SCE and MN analysis with 5'-bromodeoxyuridine (BrdU) or cytokinesis block, respectively. In parallel, the comet assay was performed with blood samples from the same persons for detection of DNA damage, including single-strand breaks (SSB) and alkali-labile sites (ALS). In workers exposed mostly to lead, blood Pb concentrations ranged from 282 to 655 microg/l, while the range in the controls was from 17 to 180 microg/l. Cd concentration in lead-exposed workers fell in the same range as for the controls. In workers exposed mainly to cadmium, blood Cd levels varied from 5.4 to 30.8 microg/l, with respective values for controls within the range of 0.2-5.7 microg/l. Pb concentrations were similar as for the controls. The incidence of MN in peripheral lymphocytes from workers exposed to Pb and Cd was over twice as high as in the controls (P<0.01). Using a combination of conventional scoring of MN and FISH with pan-centromeric probes, we assessed that this increase may have been due to clastogenic as well as aneugenic effects. In Cd- and Pb-exposed workers, the frequency of SCEs as well as the incidence of leukocytes with DNA fragmentation in lymphocytes were slightly, but significantly increased ( P<0.05) as compared with controls. After a 3h incubation of the cells to allow for DNA repair, a clear decrease was found in the level of DNA damage in the controls as well as in the exposed workers. No significant influence of smoking on genotoxic damage could be detected in metal-exposed cohorts. Our findings indicate that lead and cadmium induce clastogenic as well as aneugenic effects in peripheral lymphocytes, indicating a potential health risk for working populations with significant exposures to these heavy metals.     

Pb and Zn accumulation in a Cd-hyperaccumulator (Viola baoshanensis)

Viola baoshanensis has been identified as a Cd-hyperaccumulator, however, its ability to accumulate Pb or Zn is less certain. Therefore, this study focused on determining whether or not V. baoshanensis can accumulate Pb or Zn, by means of field survey, hydroponic and pot experiments. In addition, we also tried to obtain further information on the Cd hyperaccumulating characteristics of this species. Under field conditions, V. baoshanensis accumulated on average 1090 mg Cd kg(-1), 1902 mg Pb kg(-1) and 3428 mg Zn kg(-1) in its shoots, respectively. In hydroponic and pot experiments, V. baoshanensis showed high tolerance to Cd, Pb, and Zn, as well as the ability to accumulate exceptionally high concentrations of the three elements in its shoots (> 2% Cd, > 1% Pb, and > 0.5% Zn on a dry matter basis). These results, taken together, suggested that V. baoshanensis is not only a Cd-hyperaccumulator, but also a strong accumulator of Pb and Zn.     

Failure of antimony trioxide to induce micronuclei or chromosomal aberrations in rat bone-marrow after sub-chronic oral dosing

Antimony trioxide (Sb2O3, CAS 1309-64-4) is widely used as a flame retardant synergist in a number of household products, as a fining agent in glass manufacture, and as a catalyst in the manufacture of various types of polyester plastics. It does not induce point mutations in bacteria or mammalian cells, but is able to induce chromosomal aberrations (CA) in cultured cells in vitro. Although no CA or micronuclei (MN) have been induced after acute oral dosing of mice, repeated oral dosing for 14 or 21 days resulted in increased CA in one report, but did not result in increased MN in another. In order to further investigate its in vivo genotoxicity, Sb2O3 was dosed orally to groups of rats for 21 days at 250, 500 and 1000 mg/kg day. There were no clinical signs of toxicity in the Sb2O3-exposed animals except for some reductions in body-weight gain in the top dose group. Toxicokinetic measurements in a separate study confirmed bone-marrow exposure, and at higher levels than would have been achieved by single oral dosing. Large numbers of cells were scored for CA (600 metaphases/sex group) and MN (12,000 PCE/sex group) but frequencies of CA or MN in Sb2O3-treated rats were very similar to controls, and not biologically or statistically different, at all doses. These results provide further indication that Sb2O3 is not genotoxic to the bone marrow of rodents after 21 days of oral administration at high doses close to the maximum tolerated dose.     

Genotoxicity of the cancer chemopreventive drug candidates CP-31398, SHetA2, and phospho-ibuprofen

The genotoxic activities of three cancer chemopreventive drug candidates, CP-31398 (a cell permeable styrylquinazoline p53 modulator), SHetA2 (a flexible heteroarotinoid), and phospho-ibuprofen (PI, a derivative of ibuprofen) were tested. None of the compounds were mutagenic in the Salmonella/Escherichia coli/microsome plate incorporation test. CP-31398 and SHetA2 did not induce chromosomal aberrations (CA) in Chinese hamster ovary (CHO) cells, either in the presence or absence of rat hepatic S9 (S9). PI induced CA in CHO cells, but only in the presence of S9. PI, its parent compound ibuprofen, and its moiety diethoxyphosphoryloxybutyl alcohol (DEPBA) were tested for CA and micronuclei (MN) in CHO cells in the presence of S9. PI induced CA as well as MN, both kinetochore-positive (Kin+) and -negative (Kin-), in the presence of S9 at ≤100μg/ml. Ibuprofen was negative for CA, positive for MN with Kin+ at 250μg/ml, and positive for MN with Kin- at 125 and 250μg/ml. DEPBA induced neither CA nor MN at ≤5000μg/ml. The induction of chromosomal damage in PI-treated CHO cells in the presence of S9 may be due to its metabolites. None of the compounds were genotoxic, in the presence or absence of S9, in the GADD45α-GFP Human GreenScreen assay and none induced MN in mouse bone marrow erythrocytes.     

Uptake of lead,cadmium and zinc by the fairy shrimp,Branchinecta longiantenna (Crustacea: Anostraca)

Individuals of the fairy shrimp, Branchinecta longiantenna, were subjected to 5 concentrations (0.1 to 15 mg l^–1) of Pb in diluted habitat water at 13 °C. Lead concentrations (mg kg^–1 wet weight) in the animals were determined at 2-day intervals by digestion in nitric acid followed by atomic absorption analysis. The shrimp were also subjected to 0.1 mg l^–1 media of Cd and Zn, separately.Uptake rates by the fairy shrimp for the three metal ions at 0.1 mg l^–1 were: 0.111, 0.0885, and 0.0460 mg kg^–1 day^–1 for Zn, Pb, and Cd, respectively. After 2 days in 1.0 mg l^–1 Cd or Zn, the animals expired; but they surviced for 8 days in a 10 mg l^–1 Pb medium and for 2 days in 25 mg l^–1 Pb. Lead uptake demonstrated a linear dependence on the Pb concentration in the media.Shrimp survived at much higher tissue accumulations of Pb compared to Zn and Cd. Estimated lethal doses were 20, 1.2–2.4, and 0.4–1.4 mg kg^–1 wet weight for Pb, Zn, and Cd, respectively. Pb was found to be at much lower concentration than Cd or Zn in the natural pond water but between Cd and Zn levels in the sediment. Thus Cd and Zn probably present a greater threat to B. longiantenna than Pb, although Pb may be in higher concentration in the environment.Contribution 47, Laboratory of Ecology, The Claremont Colleges, Claremont, CA 91711, USA. Send reprint requests to Clyde Eriksen.     

Comparative evaluation of the in vitro micronucleus test and the comet assay for the detection of genotoxic effects of X-ray radiation

The genotoxic effects of X-ray radiation on human lymphocytes were measured using the single cell gel electrophoresis (SCGE) assay (comet assay) and the cytokinesis-blocked micronucleus (CBMN) test; both were carried out in vitro on isolated human lymphocytes in order to compare the relationship and sensitivity of these two detecting methods. The radiation-doses were 0.00, 0.02, 0.05, 0.10, 0.25, 0.50, 1.00 and 2.00 Gy. In the comet assay, the average comet length (38.6+/-0.8 microm) of 0.05 Gy was significantly longer than that (29.4+/-1.1 microm) of 0 Gy (P<0.01), moreover, the average comet length increased with the dose of X-ray radiation. In the CBMN, both the average micronucleus rate (MN) and micronucleated cell rate (MNC) of 0.05 Gy were 11.5+/-4.5 per thousand, which showed no difference with that (7.5+/-0.5 per thousand) of 0 Gy (P>0.05). The lowest dose, which induced significant increase of average MN and MNC, was 0.25 Gy. The average MN and MNC rates increased with radiation-dose. The results showed that there was correlation between SCGE and CBMN, and the sensitivity of SCGE was significantly higher than that of CBMN.     

Evaluation of genotoxicity, cytotoxicity and cytostasis in human lymphocytes exposed to patulin by using the cytokinesis-block micronucleus cytome (CBMN cyt) assay

Patulin (PAT) is a fungal secondary metabolite commonly present in apples and apple products. In the present study, PAT was evaluated for its genotoxic, cytotoxic and cytostatic effects to human peripheral blood lymphocytes by using the cytokinesis-block micronucleus cytome (CBMN Cyt) assay. Lymphocyte cultures were treated with PAT at the following concentrations, 0.1, 0.3, 0.5, 1.0, 2.5, 5.0, and 7.5 μM, as well as 0.5 μM mitomycin c (MMC) as a positive control and dimethyl sulfoxide (DMSO) as a vehicle control. PAT was found to induce nucleoplasmic bridges (NPBs) at 5.0 and 7.5 μM concentrations (P?<?0.05), apoptotic cells at 0.1, 1.0, 5.0 μM (P?<?0.05), 7.5 μM concentrations (P?<?0.01) and necrotic cells at 0.3 and 2.5 μM (P?<?0.05), 5.0 and 7.5 μM (P?<?0.01) concentrations in human lymphocytes. The 2.5, 5.0, and 7.5 μM PAT concentrations also led to a clear decrease in the nuclear division index (NDI) (P?<?0.05). PAT caused a significant dose-dependent increase in the number cells of NPBs, in the frequency of apoptotic and necrotic cells, and a significant dose-dependent decrease in the NDI values in lymphocytes. These results indicate that PAT at high concentrations is genotoxic, cytotoxic and cytostatic in cultured human lymphocytes.     

Changes of photosynthetic parameters in cucumber leaves under Cu,Cd, and Pb stress

The effects of Cu, Cd, and Pb toxicity on photosynthesis in cucumber leaves (Cucumis sativus L.) were studied by the measurements of gas exchange characteristics, chlorophyll (Chl) fluorescence parameters, and Chl content. Concentrations of metals in sequence of 20 μM Cu, 20 and 50 μM Cd, and 1 000 μM Pb decreased the plant dry mass to 50–60 % after 10 d of treatment whereas 50 μM of Cu decreased it to 30 %. The content of Cd in leaves of plants treated with 50 μM Cd was three times higher than the contents of Cu and Pb after plant treatment with 50 μM Cu or 1 000 μM Pb. Hence Cd was transported to leaves much better than Cu and Pb. Nevertheless, the net photosynthetic rate and stomatal conductance in leaves treated with 50 μM Cu or Cd were similarly reduced. Thus, Cu was more toxic than Cd and Pb for photosynthesis in cucumber leaves. None of the investigated metals decreased internal CO₂ concentrations. Also the effect of metals on potential efficiency of photosystem 2, PS2 (F_v/F_m) was negligible. The metal dependent reduction of PS2 quantum efficiency (Φ_PS2) after plant adaptation in actinic irradiation was more noticeable. This could imply that reduced demand for ATP and NADPH in a dark phase of photosynthesis caused a down-regulation of PS2 photochemistry. Furthermore, in leaves of metal-treated plants the decrease in water percentage as well as lower contents of Chl and Fe were observed. Thus photosynthesis is not the main limiting factor for cucumber growth under Cu, Cd, or Pb stress.     

Distribution and contamination pattern of heavy metals from surface sediments in the southern part of Caspian Sea,Iran

This study concentrates on the speciation and distribution patterns of some heavy metals (Pb, Ni, Cd, Zn, and Cu) in surface sediments in the southern part of the Caspian Sea, the biggest lake in the world, to obtain an overall classification for the origins of metals in the area using a sequential extraction technique. At all sampling stations, Pb, Ni, Zn, and Cu were mostly (>50%) accumulated in the resistant fraction, which indicated that there were no significant anthropogenic inputs of Pb, Ni, Zn, and Cu into the surface sediments of the south Caspian Sea. Guilan province on the west coast of Caspian Sea accumulated higher percentages of non-resistant fractions of Pb and Zn, while Mazandaran and Golestan provinces in the middle and western parts of the Caspian Sea, in the Iranian zone, accumulated higher percentages of non-resistant fractions of Ni and Cu. The present study revealed that the coastal area of the south Caspian Sea is still not seriously contaminated. Cadmium in Guilan and Golestan provinces were dominated by non-resistant fractions (55–69%), which indicated more anthropogenic inputs of Cd on the south coast of the Caspian Sea in comparison with other metals.     

Leaching of Zinc,Cadmium, and Lead in a Sandy Soil Due to Application of Poultry Litter

Dissolved organic matter in poultry litter could contribute organic ligands to form complexes with heavy metals in soil. The soluble complexes with heavy metals can be transported downward and possibly deteriorate groundwater quality. To better understand metal mobilization by soluble organic ligands in poultry litter, soil columns were employed to investigate the movement of zinc (Zn), cadmium (Cd), and lead (Pb). Uncontaminated soil was amended with Zn, Cd, and Pb at rates of 400, 8, and 200 mg kg ^{? 1} soil, respectively. Glass tubes, 4.9-cm-diameter and 40-cm-long, were packed with either natural or metal-amended soil. The resulting 20-cm-long column of soils had bulk density of about 1.58 g cm ^{? 3} . Columns repacked with natural or amended soil were leached with distilled water, 0.01 M EDTA, 0.01 M CaCl ₂ , or poultry litter extract (PLE) solutions. Low amounts of Zn, Cd, and Pb were leached from natural soil with the solutions. Leaching of Zn, Cd, or Pb was negligible with distilled water. In the metal-amended soil, EDTA solubilized more Zn, Cd, and Pb than CaCl ₂ and PLE. The breakthrough curves of Zn and Pb in the PLE and CaCl ₂ were similar, indicating they have similar ability to displace Zn and Pb from soils. Compared with Zn and Cd the PLE had a small ability to solubilize Pb from metal-amended soil. Thus, the application of poultry litter on metal-contaminated soils might enhance the mobility of Zn and Cd.     

Ion exchange during heavy metal bio-sorption from aqueous solution by dried biomass of macrophytes

In this study, potentials of oven dried biomass of Eichhornia crassipes, Valisneria spiralis and Pistia stratiotes, were examined in terms of their heavy metal (Cd, Ni, Zn, Cu, Cr and Pb) sorption capacity, from individual-metal and multi-metal aqueous solutions at pH 6.0+/-0.1 (a popular pH of industrial effluent). V. spiralis was the most and E. crassipes was the least efficient for removal of all the metals. Cd, Pb and Zn were efficiently removed by all the three biomass. Cd was removed up to 98% by V. spiralis. Sorption data for Cr, Ni and Cd fitted better to Langmuir isotherm equation, while, the sorption data for Pb, Zn and Cu fitted better to Freundlich isotherm equation. In general, the presence of other metal ions did not influence significantly the targeted metal sorption capacity of the test plant biomasses. Ion exchange was proven the main mechanism involved in bio-sorption and there was a strong ionic balance between adsorbed (H(+) and M(2+)) to the released ions (Na(+) and K(+)) to and from the biomass. No significant difference was observed in the metal exchanged amount, by doubling of metal concentration (15-30 mg/l) in the solution and employing individual-metal and multi-metal solutions.     

Lead, zinc and cadmium accumulation from two metalliferous soils with contrasting calcium contents in heavy metal-hyperaccumulating and non-hyperaccumulating metallophytes: a comparative study

Aims and background

We previously compared metallicolous (M) and non-metallicolous (NM) populations of Noccaea (=Thlaspi) caerulescens, Silene vulgaris, and Matthiola flavida for their abilities to tolerate and (hyper)-accumulate lead (Pb) in hydroponics. In the present study we aimed 1) to check the hyperaccumulation and tolerance abilities of these populations in controlled experiments using metalliferous soils, 2) to test the M. flavida M population for Zn and Cd hypertolerance in hydroponics.

Methods

Plants were grown in hydroponics and fertilized metalliferous substrates, collected from a Zn/Pb smelter sinter deposit near Plombières, Belgium (low pH, low Ca), and a tailing of the Irankouh Zn/Pb mine, Iran (high pH, high Ca). Metal tolerance was assessed from root growth inhibition in hydroponics, or mortality, stunting or chlorosis in the experiments with soil.

Results

Metallicolous M. flavida did not show hypertolerance or hyperaccumulation of Cd or Zn in hydroponics. Only one of the N. caerulescens M populations and the native S. vulgaris M population were able to grow in Plombières soil, whereas the others stopped growing or died within 40?days. All the populations survived and maintained growth for 40?days in Irankouh soil. When grown in Irankouh soil, the M population of M. flavida hyperaccumulated Pb. N. caerulescens hyperaccumulated Zn from Plombières soil, but not from Irankouh soil.

Conclusions

The M. flavida M population is non-Pb-hypertolerant. It hyperaccumulates Pb from Irankouh soil, but not from Pb-amended nutrient solution. N. caerulescens does not hyperaccumulate Zn from the calcareous Irankouh soil.     

Induction of metallothionein-like proteins in the digestive gland of Mytilus galloprovincialis after a chronic exposure to the mixture of trace heavy metals

Abstract

The amount of metallothionein-like proteins (MTLPs) induced in the digestive gland of exposed and control specimens of Mytilus galloprovincialis is measured after the 14 days of the experiment. Three groups of mussels were kept in a flow-through seawater. The amount of induced MTLPs has been measured in the digestive gland of mussels exposed to the added Cd and, separately, to the added mixture of Cd, Cu, Pb while Zn was for all three experimental groups enhanced to 11.80 μg L^?1 in flow-through seawater, as the consequence of the contamination from the pipeline for bringing the seawater to the aquarium. The induced amount of MTLPs in exposed mussels is compared with its amount in control specimens, which were kept for 14 days in the flow-through seawater at the natural concentration level of Cd, Cu, Pb and enhanced Zn concentration, as previously explained. The metal concentrations in seawater to which mussels were exposed for 14 days were of Cu 3 times, of Pb 7 times, of Zn 20 times higher than their natural concentration level. Only the concentration of Cd was 100 times higher than the natural one. Therefore, the presented results are applicable to understanding the effect of the mixture of environmentally relevant metals on the induced amount of MTLPs as the indicator substance of seawater pollution.