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Heavy metal induced protein synthesis in fish cell lines.

The regulation of heavy metal induced gene expression was investigated in two fish cell lines: the rainbow trout hepatoma (RTH) and Chinook salmon embryo (CHSE) cells. The induction of metallothionein (MT) synthesis occurred in RTH cells exposed to zinc, and to a lesser extent, following exposure to cadmium. The time courses of MT synthesis were different for the different metal inducers, suggesting that MT may be differentially regulated in these cells. CHSE cells, unlike RTH cells, did not synthesize MT in response to metal treatment. Treatment of the cells with 5-azacytidine prior to heavy metal exposure resulted in the induction of MT synthesis. Since these cells were embryonic in origin, these findings raised the possibility that MT gene expression may be developmentally regulated in fish. Analysis of the entire spectrum of cellular proteins has revealed the synthesis of an unknown, 14,000 dalton, metal-inducible protein (MIP) and various stress proteins following exposure of fish cells to heavy metals.

Animals↗

Potential of weed species applied to remediation of soils contaminated with heavy metals.

To screen out a series of ideal plants that can effectively remedy contaminated soils by heavy metals is the main groundwork of phytoremediation engineering and the first step of its commercial application on a large scale. In this study, accumulation and endurance of 45 weed species in 16 families from an agricultural site were in situ examined by using the pot-culture field experiment, and the remediation potential of some weed species with high accumulation of heavy metals was assayed. The results showed that Solanum nigrum and Conyza canadensis can not only accumulate high concentration of Cd, but also strongly endure to single Cd and Cd-Pb-Cu-Zn combined pollution. Thus 2 weed species can be regarded as good hyperaccumulators for the remediation of Cd-contaminated soils. Although there were high Cd-accumulation in Artemigia selengensis, Znula britannica and Cephalanoplos setosum, their biomass was adversely affected due to action of heavy metals in the soils. If the problem of low endurance to heavy metals can be solved by a reinforcer, 3 weed species can be perhaps applied commercially.

Biodegradation, Environmental↗

In vitro and in vivo effects of heavy metals on mussel digestive gland hexokinase activity: the role of glutathione.

Hexokinase (E.C. 2.7.1.1), the enzyme responsible for glucose phosphorylation to G-6P, is inactivated by SH reagents and oxyradicals, and its inhibition has been involved in heavy metal toxicity in mammalian systems. In this work, the possibility that hexokinase activity could be affected by both heavy metal binding and oxidative stress conditions also in mussel tissues (Mytilus galloprovincialis Lam.) was investigated. The results obtained in vitro demonstrate that heavy metals inhibited digestive gland hexokinase (with Cd2+ > Cu2+ > Hg2+ > Zn2+ > Pb2+) and suggest a role for GSH in the protection against the heavy metal effects. Hexokinase activity was also reduced by addition of iron/ascorbate, indicating a susceptibility of the enzyme to metal-mediated oxyradical production. The effects of Cu2+ treatment (3 days, 40 micrograms l-1 per animal) on hexokinase activity and on the GSH/GSSG status were then evaluated in mussels exposed to a cycle of air exposure/reimmersion. In Cu-exposed mussels, a significant decrease in hexokinase activity and a parallel reduction in tissue GSH levels were observed, suggesting that the two effects of metal treatment could be related; however, hexokinase activity progressively recovered during air exposure and reimmersion, whereas the level of GSH showed a further decrease during air exposure followed by recovery after reimmersion. The in vitro results therefore indicate that mussel digestive gland hexokinase is susceptible to inactivation by heavy metal binding and suggest a role for GSH in the protection against the effects of heavy metals. The effects of copper were confirmed by the results obtained in vivo. The possible relationship between hexokinase activity and the level of GSH in the digestive gland of control and Cu-exposed mussels during air exposure and reimmersion are discussed, taking into account the balance between pro-oxidant and antioxidant processes at different stages of exposure.

Animals↗

Process evaluation for optimization of EDTA use and recovery for heavy metal removal from a contaminated soil.

This study aimed to establish an optimized, closed loop application of ethylenediaminetetraacetic acid (EDTA) in heavy metal removals from a contaminated soil through integrating EDTA recovery/regeneration and metal precipitation processes in the treatment train. Three divalent heavy metals were investigated, namely, Pb, Cd, and Ni. The extractability of the metals by EDTA followed the decreasing order of CdPb>>Ni. The first part of this study was to search for the optimal use of the fresh EDTA in removing these heavy metals from the contaminated soil. The second part of this study was devoted to the recovery/regeneration of the spent EDTA which followed the sequential processes involving (1) complex destabilization by adding ferric ion (Fe(III)) to liberate Pb, Cd, and Ni, (2) precipitation of the liberated Pb, Cd, and Ni in phosphate (PO4(3-)) forms, and (3) precipitation of the excess Fe(III) which eventually produced free EDTA for reuse. The process variables were dosages of Fe(III) and PO4(3-), pH and reaction times. Laborious trial experiments would be needed in searching for the optimum conditions for the above processes. To expedite this exercise, a geochemical equilibrium model, MINTEQA2, was used to find the thermodynamically favorable conditions for recoveries of both EDTA and heavy metals. This was then followed by experimental examination of the process kinetics to observe for the optimal reaction time for each thermodynamically favorable process. This study revealed that 2 h of reaction time each for the complex destabilization reaction and the metal phosphate precipitation reaction was sufficient to achieve equilibrium. With the optimized process condition identified in this study, a total of 95%, 89% and 90% of the extracted Pb, Cd and Ni, respectively, could be precipitated from the spent EDTA solution, with 84% EDTA recovery. The reused EDTA maintained more than 90% of its preceding extraction power in each cycle of reuse.

Chelating Agents↗

Individual metal responsive elements of the human metallothionein-IIA gene independently mediate responses to various heavy metal signals.

Metallothioneins (MTs) are small metal-binding proteins that have a role in the defense against heavy metals. Mammalian MT genes are transcriptionally activated by metals such as Cd and Zn through multiple copies of the metal responsive element (MRE) present in the 5'-flanking region. To examine whether each MRE in a single promoter has a distinct role, we characterized seven MREs located upstream of the human MT-IIA gene. By transient transfection experiments using MRE-driven reporter gene constructs, individual MREs were assayed for the activity to mediate transcription in response to several heavy metal species. Four MREs including MREs a, b, e and g independently mediated reporter gene expression in response to Zn, Cd and Hg, while other MREs were not responsive to any of these metals. These results suggest that the multiplicity of MRE contributes to enhancing its activity, rather than providing functional diversity.

DNA-Binding Proteins↗

Effects of heavy metals on the Ca(2+)-ATPase activity present in gill cell plasma-membrane of mussels (Mytilus galloprovincialis Lam.).

1. Heavy metals (Hg2+, Cu2+, Cd2+, Zn2+, Pb2+) at micromolar concentrations strongly inhibit the Ca(2+)-ATPase activity present in the plasma-membrane obtained from the gill cells of Mytilus galloprovincialis Lam. Heavy metals act through inhibition of the formation of the phosphorylated intermediate. 2. All the heavy metals tested inhibit the Ca(2+)-ATPase activity, the effect following the order: Hg2+ > Pb2+ > Cu2+ > Cd2+ > Zn2+; the simultaneous addition of different heavy metals causes a summatory inhibition of the enzyme activity; addition to the reaction mixture of GSH at a final concentration of 0.5 mM, reverses inhibitory effects of heavy metals. 3. The inhibitory effects of Cu2+ on Ca(2+)-ATPase are highly enhanced by addition of ascorbate to the reaction mixture. In the presence of ascorbate (100 microM), copper strongly stimulates the lipid peroxidation damage of the gill plasma-membranes, a result that may explain the high copper cytotoxicity.

Animals↗

Speciation of heavy metals in marine sediments vs their bioaccumulation by mussels.

Total contents and speciation of selected heavy metals, including Al, Fe, Co, Ni, Pb, Zn, Cu, Cr, were measured in sediment samples and mussels Mya arenaria and Astarte borealis collected in the Horsund Fjord off Spitsbergen (Norwegian Sea) and the Bay of Gdansk (Baltic Sea). The investigation aimed at revealing differences in the accumulation pattern of heavy metals in mussels inhabiting sediments characterized by varying metal bioavailability. The contents of metals adsorbed to sediments and associated with iron and manganese hydroxides, which were obtained by sequential extraction, were utilized as a measure of metal bioavailability. The contents of Cd, Pb, Zn, Cu and Cr in mussels collected off Spitsbergen were generally lower than those in mussels from the Baltic Sea. In sediments collected off Spitsbergen the bioavailable fraction represented a small proportion (0-3.7% adsorbed metals and 0-11% associated with metals hydroxides) of total heavy metal contents. In sediments from the Baltic Sea the percentages of metals adsorbed and bound to hydroxides were 1-46% and 1-13%, respectively. The differences in bioavailable metal contents measured in sediments were utilized to explain the different contents of metals in mussels collected in the corresponding sites.

Adsorption↗

Engineering plant-microbe symbiosis for rhizoremediation of heavy metals.

The use of plants for rehabilitation of heavy-metal-contaminated environments is an emerging area of interest because it provides an ecologically sound and safe method for restoration and remediation. Although a number of plant species are capable of hyperaccumulation of heavy metals, the technology is not applicable for remediating sites with multiple contaminants. A clever solution is to combine the advantages of microbe-plant symbiosis within the plant rhizosphere into an effective cleanup technology. We demonstrated that expression of a metal-binding peptide (EC20) in a rhizobacterium, Pseudomonas putida 06909, not only improved cadmium binding but also alleviated the cellular toxicity of cadmium. More importantly, inoculation of sunflower roots with the engineered rhizobacterium resulted in a marked decrease in cadmium phytotoxicity and a 40% increase in cadmium accumulation in the plant root. Owing to the significantly improved growth characteristics of both the rhizobacterium and plant, the use of EC20-expressing P. putida endowed with organic-degrading capabilities may be a promising strategy to remediate mixed organic-metal-contaminated sites.

Bacterial Proteins↗

[Distribution of heavy metals in human ejaculate--a histochemical study].

With the help of a modified dithizone method, heavy metals could be demonstrated in the sperms, the ejaculate fluid, and the spermatogenic cells of men. The heavy metals were localized in the sperms in form of small red granules particularly in the mid piece with a distribution like a "string of pearls". The sperms with a lower motility index often did not show any heavy metal granules in the mid pieces. The absence of essential heavy metals like zinc, iron or manganese may lead to the inhibition of enzyme systems in the mid pieces of the sperms, which are essential for the motility.

Dithizone↗

Heavy metals in sediments from the Paraguay River: a preliminary study.

The first results ever obtained on heavy metal concentrations (Fe, Mn, Cr, Cu, Zn and Pb) in the Paraguay River surface sediments are presented. Samples collected at 11 locations, along a distance of 570 km, between the cities of Bahia Negra and Alberdi in Paraguay, for six different periods between November 1991 and 1993, were analyzed. The Paraguay sediments appeared to have features of an unpolluted river even though significant amounts of domestic and industrial effluents are discharged near the river channel. The relative heavy metal enrichment in sediments between Bella Vista and Asunción, caused by local domestic sewage and industrial outfalls, is less than for the shale standard values. The heavy metal content of the sediments exhibited seasonal variations. Enhanced organic matter content and biochemical oxygen demand of the river load in winter caused most likely a retainment of the heavy metals in a dissolved state. Consequently, the sediments deposited in the winter were relatively depleted in these elements.

Environmental Monitoring↗

Heavy metal fates in laboratory bioretention systems.

Key to managing heavy metals in bioretention is to understand their fates in bioretention facilities. In this study, pot prototypes filled with bioretention media were built to simulate the conditions of natural growth of plants. Synthetic runoff with different heavy metal loadings (copper, cadmium, lead, and zinc) was periodically applied. Metal accumulations in tissues of grasses -Panicum virgatum, Kentucky-31, and Bromus ciliatus, were investigated after 230d of growth and multiple runoff treatment events. After 183d of periodic runoff application, the concentrations of Zn, Cu, Pb and Cd with low and high loadings had the same trends in the plant tissues, Zn>Cu>Pb>Cd, following the trend of the input metal concentrations. The fates of input metals were 88-97% captured in soil media, 2.0-11.6% not captured by bioretention media, and 0.5-3.3% accumulated in plants. Compared to the metals retained by the soil, the percentages of input metals taken up by plants were relatively low due to the low plant biomass produced in this study. Greater biomass density would be required for the vegetation to have a valuable impact in prolonging the lifetime of a bioretention cell.

Metals, Heavy↗

Partitioning characteristics of targeted heavy metals in IZAYDAS hazardous waste incinerator.

Partitioning of eight targeted heavy metals (Cr, Mn, Cu, Pb, Sn, Co, Ni and Zn) was carried out during five trial burns in Izmit hazardous and clinical waste incinerator (IZAYDAS). Metal contents of the original wastes and their concentration in the bottom ash (BA), fly ash (FA), filter cake (FC) and flue gas were determined. Partitioning behavior of metals during the two-stage incineration was evaluated with respect to physico-chemical properties of feed waste and metals, and the operational conditions. Results suggest that combustion temperatures and retention times are the dominant parameters determining the volatility of metals in the first combustion chamber. Targeted metals were generally partitioned in the rank of bottom ash, filter cake, fly ash and flue gas. High filter cake/fly ash ratios showed that high temperatures in the second stage increase both the formation of gaseous metallic compounds and the enrichment of metals in fine particles. Since ESP could not be effective in removing fine particles and volatilized metallic compounds, the necessity of an additional system that would remove heavy metals efficiently was emphasized for the modern incinerators.

Gases↗

Impact of industrial waste water effluents on mycoflora of the shore sediments of the 3rd oxidation pond, with reference to biosorption of heavy metals.

The third oxidation pond at 10th of Ramadan desert receives a number of industrial waste water effluents contaminated with the heavy metal ions Zn, Cd, Cu and Ni. The species diversity and fungal community structure of seven different sites at the onshore sediments and offshore were studied. Mycological analysis resulted in isolation of 3912 fungal colonies, 11.7% of this count were recovered from the onshore sediment sites (4 sites) whereas 88.3% were from the offshore sites (3 sites), in the desert. Fungal counts and species diversity at the onshore sites tend to increase with increasing distance far from the waste water input. A complete accordance was observed among the total fungal counts and species variabilities with organic matter content at each sampling site. This relationship was reversed in case of heavy metal contents with both counts and diversity. Seventeen fungal species belonging to seven genera were isolated from all sites under study. Aspergillus spp. constituted the majority of the isolates (51.7% of the total isolates), followed by Curvularia, Cephalosporium, and Humicola. Of nine isolated Aspergillus spp., A. humicola was the most dominant (37.4% of the total catch) and appeared at all polluted sites. Therefore, A. humicola was chosen to investigate its potential for heavy metals sorption from the contaminated waste water effluent. Four days old biomass pellets could sorb a large amount of heavy metals according to the following sequence: Zn>Cd>Cu>Ni ions. Agitation significantly increased Zn and Cd sorption, but not Cu and Ni. Heavy metals sorption took place at a wide pH range and particularly increased at alkaline pH (8-9).

Aspergillus↗

[The resistance of soil microorganisms to soil pollution by heavy metals].

Microbial communities of grey podzolized soil of Left-Bank Ukraine are characterized by high potential resistance to pollution with heavy metals: above 40% of organotrophic microorganisms are capable to resist to pollution with a mixture of heavy metals (Cu2+, Cd2+, Pb2+, Zn2+) in a dose of 20 MPC of each metal. Relative amount of resistant microorganisms in polluted soil increases up to 93%. White zeolites and biohumus are promising for reclamation of soils polluted with heavy metals. They decrease toxic effect of heavy metals on soil microflora and promote development of microflora resistant to pollution.

Colony Count, Microbial↗

[Study on heavy metals in growing area soils and young fruit of Citrus grandis 'Tomentosa'].

OBJECTIVE: To determine the status of heavy metals contamination in growing area soils and young fruits of Citrus grandis. METHOD: Application of Nemerow's index for heavy metals assessment of soil and young fruit of Citrus grandis, as well as according to national environmental quality standard for soils and green trade standards of importing & exporting medicinal plants & preparations. RESULT: The heavy metals content of most of the growing area soils of Citrus grandis rates as first, and of the rest ones rates as second. The heavy metals content in young fruits of Citrus grandis is within the limitation of quantity of criteria. CONCLUSION: The environmental quality of growing area soils of Citrus grandis in Huazhou City is totally in good.

Arsenic↗

Evaluation of heavy metal inhibition of activated sludge by TTC and INT-electron transport system activity tests.

TTC and INT-electron transport system activity tests were compared for assessing heavy metal inhibition of activated sludge. The median inhibitory concentrations (IC50s) of Cu2+, Zn2+, Cd2+, Hg2+, Ni2+, Pb2+ and Ag+ measured via TTC test were lower than those measured via INT test, which indicates that the INT-electron transport system activity test was less sensitive to heavy metal toxicity than the TTC test. Tested heavy metals brought about similar decrease in TTC-electron transport system activity and COD removal rate, but less decrease in INT-electron transport system activity than COD removal rate, which suggests that the TTC-electron transport system activity is a better parameter for reflecting heavy metal inhibition of activated sludge than INT-electron transport system activity. The ranking of tested heavy metals in order of decreasing toxicity based on TTC test was Hg2+, Cd2+, Cu2+, Ag+, Zn2+, Ni2+ and Pb2+, and the ranking based on INT test was Hg2+, Ag+, Cu2+, Cd2+, Zn2+, Ni2+ and Pb2+.

Bacteria↗

Heavy metal stress reduces the deposition of calcium oxalate crystals in leaves of Phaseolus vulgaris.

Calcium oxalate (CaOx) crystals in plants may serve as a sink for the absorption of excess calcium, and they could play an important role in heavy metal detoxification. In this study, the effect of heavy metals and different calcium concentrations on the growth of calcium oxalate crystals in leaves of Phaseolus vulgaris was investigated. Different analytical techniques were used to determine the influence of exogenous lead and zinc on CaOx deposition and to detect a presence of these metals in CaOx crystals. We found a positive correlation between the calcium concentration in the nutrient medium and the production of calcium oxalate crystals in leaves of hydroponically grown plants. On the other hand, addition of the heavy metals to the nutrient medium decreased the number of crystals. Energy dispersive X-ray spectrometry did not detect the inclusion of heavy metals inside the CaOx crystals. Our investigation suggests that CaOx crystals do not play a major role in heavy metal detoxification in P. vulgaris but do play an important role in bulk calcium regulation.

Calcium Oxalate↗

[Advances in the research of genetic engineering of heavy metal resistance and accumulation in plants].

Using plants to remove or inactivate heavy metal pollutants from soils and surface waters provide a cheap and sustainable approach of Phytoremediation. However, field trials suggested that the efficiency of contaminant removal using natural hyperaccumulators is insufficient, due to that many of these species are slow growing and produce little shoot biomass. These factors severely constrain their potential for large-scale decontamination of polluted soils. Moreover, both the micronutrient and toxic metal content accumulated in crops determine the quality and safety of our food-chain. By a transgenic approach, the introduction of novel genes responsible for hyperaccumulating phenotype into high biomass plants and/or stable crops uptaking minerals as food is a promising strategy for the development of effective techniques of phytoremediation and improvement of nutritional value of stable food through a viable commercialization. Recently, the progress at molecular level for heavy metal uptaking, detoxification and hyperaccumulation in plants, and also the clarification of some functional genes in bacteria, yeasts, plants and animals, have advanced the research on genetic engineering plants of heavy metal resistance and accumulation, and on the functional genes (e . g. gsh1, MerA and ArsC) and their genetic transformated plants. These studies demonstrated commercialization potentials of phytoremediation. In this paper, the molecular approach, effects and problems in gene transformation were discussed in details, and also the strategy and emphases were probed into the future research.

Biodegradation, Environmental↗