Estimation of Cd,Pb, and Zn Bioavailability in Smelter-Contaminated Soils by a Sequential Extraction Procedure

Chemical fractionation methods may be capable of providing an inexpensive estimate of contaminant bioavailability and risk in smelter-contaminated soil. In this study, the relationship between metal fractionation and methods used to estimate bioavailability of these metal contaminants in soil was evaluated. The Potentially BioAvailable Sequential Extraction (PBASE) was used for Cd, Pb, and Zn fractionation in 12 soils contaminated from Pb and Zn mining and smelting activities. The PBASE procedure is a four-step sequential extraction: extraction 1 (E1) is 0.5 M Ca(NO₃)₂, E2 is 1.0 M NaOAc, E3 is 0.1 M Na₂EDTA, and E4 is 4 M HNO₃. Metal bioavailability for two human exposure pathways, plant uptake (phytoavailability) and incidental ingestion (gastrointestinal, Gl, availability), was estimated using a lettuce (Lactuca sativa L.) bioassay and the in vitro-Gl Physiologically Based Extraction Test(PBET). Metal in the PBASE E1 fraction was correlated with lettuce Cd (P < 0.001) and Zn (P < 0.05) and was the best predictor of Cd and Zn phytoavailability. Only total metal content or the sum of all PBASE fractions, ΣE_1–4, were correlated (P < 0.001) with PBET gastric phase for Pb. The sum of the first two PBASE fractions, ΣE_1–2, was strongly correlated (P < 0.001) with Pb extracted by the PBET intestinal phase. The PBASE extraction method can provide information on Cd and Zn phytoavailability and Gl availability of Pb in smelter-contaminated soils.     

Oxalate Extraction of Pb and Zn from Polluted Soils: Solubility Limitations

Oxalate (Ox) was used to extract Pb and Zn from industrially contaminated soils. Although Ox effectively releases metals bound by hydrous oxide soil components, it forms insoluble salts with some heavy metals unlike conventional extractants (e.g., EDTA). The insolubility of PbOx(s) (K_sp=2.74 × 10^?11) precluded the use of Ox as a single-step extractant even for soils mildly contaminated with Pb. The usefulness of Ox as a Zn extractant, however, depends on the level of soil contamination. A Zn solubility model, based on published equilibrium constants, was developed to assess Ox suitability as a function of system conditions. Precipitation of ZnOx(s) hindered Zn recovery under acidic conditions where formation of soluble oxalato complexes was small. For pH<3, the presence of 1?M Ox actually reduced Zn release compared to simple acid washing. Although Ox displaces oxide-bound metals and thus is potentially useful in soil washing, solubility limitations must be defined for effective remediation of metal-laden soils.     

Distribution patterns of Cd,Cu, Mn,Pb and Zn in wood fly ash emitted from domestic boilers

Abstract

Wood fuels being a renewable source of primary energy have been considered environmentally friendly. However, wood combustion in domestic boilers is a source of air pollution. The lack of a dust collection device is the reason why flue gases emit a significant load of particulate pollutants into the air, including heavy metals. The aim of this research was to assess the environmental hazard caused by both emissions of heavy metals during wood combustion in domestic boilers and their chemical forms present in fly ash.

From the various wood fuels burnt in domestic boilers, the fly ash selected for this study came from the combustion of briquettes of softwood from non-polluted areas, and from burning hardwood fuel from trees exposed to pollutants from heavy traffic. The wood fuels satisfy the quality demands determined in the EN 14961 Solid Biofuels - Fuel quality assurances. However, the concentrations of Cd, Cu, Pb and Zn in the fly ash are considerably higher than the appropriate limit values determined for soil improvers. Sequential extraction shows that Cd and Zn are associated mainly with the water leaching and carbonate fractions, regarded as mobile and bioavailable, and pose the potentially greatest hazard to the environment and human health. Cu, Mn and Pb associated with less mobile fractions may not pose a direct air quality hazard but, due to their high concentrations, medium-term and long-term effects on soils and surface and subsurface waters should be considered.     

Distribution of Toxic Trace Metals Zn,Cd, Pb,and Cu in Tirupati Soils,India

The concentrations of toxic trace metals in the soil samples collected from Tirupati, India, have been determined using differential pulse anodic stripping voltammetry (DPASV). The total metal concentrations of the soils in the study area were in the following ranges: 19.5 to 23.6 mg of Zn, 0.032 to 0.036 mg of Cd, 15.8 to 18.9 mg of Pb, and 19.0 to 23.4 mg of Cu per kg soil. Analysis of standard reference material IAEA-SOIL-5 indicates good accuracy. Recoveries were nearly quantitative for all elements studied. Comparison of the average metal concentration levels with world averages indicates an elevated value for Pb. The applicability of this method was crosschecked with AAS and the results were in good agreement.     

Epidermal uptake of Pb,Cd, and Zn in tubificid worms

Summary The epidermal uptake of Pb, Cd, and the essential element Zn by tubificid worms was investigated. The animals were starved and contaminated via the water column at 4°C and 25°C. Atomic absorption spectroscopy (AAS), energy-dispersive X-ray microanalysis (EDX), laser induced mass analysis (LAMMA), electron microscopy, and the sulfide-silver method at the electron microscopical level were used. The sulfidesilver method revealed many reaction products indicating heavy metals in membrane-limited lysosomal structures in epidermal cells of contaminated Limnodrilus udekemianus. In these lysosomes, which were particularly abundant in the epidermis of the hind end, Pb, Cd, and Zn were detected by LAMMA and EDX analysis. Beside this distinct occurrence of the three elements a uniform pattern of reaction products was found in epidermal cells after contamination with Cd and Zn, but not after treatment with Pb. As shown by atomic absorption spectroscopy, Cd was the most enriched element at 25°C, followed by Pb and Zn. Simultaneous application of Zn reduced the uptake of Cd, whereas the uptake of Pb was increased. At 4° C uptake of Pb and Cd was slower than at 25° C, while Zn uptake was blocked completely. This shows that heavy metal uptake can be strongly dependent on environmental conditions, which has to be taken into account if animals are used as monitor organisms for heavy metal pollution in the environment.     

Investigation of Vetiver Grass Capability in Phytoremediation of Contaminated Soils with Heavy Metals (Pb,Cd, Mn,and Ni)

ABSTRACT

Soil contamination with heavy metals has become a worldwide concern. A sustainable technology to mitigate heavy metal contamination is extremely important. Phytoremediation is a cost-effective method, environmentally friendly, and esthetically pleasing. The aim of this study was to investigate the potential of Vetiver phytoremediation of soils contaminated with heavy metals. This research was conducted as a factorial design with four different heavy metals (lead, cadmium, manganese, and nickel) with three varying levels and also three replications for each treatment. Statistical analysis of data was performed using SPSS19 software and analysis of variance, Duncan and Pearson correlation tests. The results showed that, the highest uptake rate was related to lead metal with 282.45 mg/kg of dry soil and 83.4% uptake percentage. Then, the mean and percentage of adsorption for cadmium, nickel and manganese were 248.3 mg/kg (53.2%), 69.4 mg/kg (65.5%), and 63.29 mg/kg (61%), respectively. Lead was found to be the main component of uptake by Vetiver plant. It was found that the roots of the plant have absorbed more heavy metals than the shoots. And at the roots in total 1089.05 and on average 363.01 mg/kg and at the shoots 901.19 and on average 300.39 mg/kg, the metals used were adsorbed on three levels and four treatments. The results of analysis of variance, Duncan test and Pearson correlation showed that the effect of applied treatments on lead uptake in roots and shoots increased significantly (P ≤ 0.05) with increasing levels of treatments. The biological concentration factor was more than one, and the transfer factor was close to one. Therefore, it can be used as a phytostablization plant. The results showed that Vetiver can be considered as a refining plant due to its vegetative characteristics, cost-effectiveness and high adaptation to environmental conditions.     

Phytoremediation of Cd, Cr, Cu, Mn, Fe, Ni, Pb and Zn from aqueous solution using Phragmites cummunis, Typha angustifolia and Cyperus esculentus

A comparative bioaccumulation pattern and ultra structural changes were studied in Phragmites cummunis, Typha angustifolia and Cyperus esculentus in mixed metals solution of cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), lead (Pb) and zinc (Zn). P. cummunis was observed to be a shoot accumulator for Cr, Fe, Mn, Ni, Pb, and Zn. However, T. angustifolia was found to be a root accumulator for Cd, Cr, Cu, Fe, Ni and Pb. In addition, C. esculentus also accumulated most of the tested heavy metals in the roots, while Mn and Fe were translocated up to leaves. Further, the long term metal treatment showed maximum accumulation of all heavy metals in P. cummunis followed by T. angustifolia and C. esculentus. Among heavy metals, Fe was accumulated maximum, i.e., >1000 microg g(-1) by all three plants. Simultaneously, the adverse effects on biochemical parameters were noted earlier in C. esculentus than T. angustifolia and P. cummunis. Ultra structural observation showed the cellular changes in wetland plants after longer exposure. Results revealed that P. cummunis and T. angustifolia had more potential for tested metals than C. esculentus. This study established that these wetland plants could be used for heavy metals phytoremediation from metal containing industrial wastewater.     

Leaching of Cd,Zn, Pb,and Cu in Packed and Undisturbed Columns of Soils Affected by the Spill from a Pyrite Mine in the South of Spain

A study has been made of the leaching of Cd, Zn, Pb, and Cu in three representative soils within the zone affected by the spill from a pyrite mine in Aznalcollar (Sevilla, Spain) employing packed soil columns. According to the breakthrough and cumulative leaching curves, the relative mobilities of the different toxic elements in the columns are as follows: Cd> Zn> Cu> Pb. The effect of leaching on the distribution of metals as a function of depth using intact soil cores was also studied. The results showed that the soils themselves have a good capacity for immobilizing the soluble fraction of the elements from the spilled mud. This capacity varied as follows: clayey soil with a high carbonate content > clayey soil with a moderate carbonate content > sandy-clay loam soil with a low carbonate content. However, sandy soils with a low carbonate content could pose a risk to groundwater if initial contamination was high. These results could be considered during the evaluation of remedial technologies for the immobilization of soil metals.     

Interspecific differences in Zn,Cd and Pb accumulation by freshwater algae and bryophytes

The relationships between the concentrations of zinc, cadmium and lead in aquatic plants and the concentrations of these metals in the ambient water have been compared for three algae (Lemanea fluviatilis, Cladophora glomerata, Stigeoclonium tenue), one liverwort (Scapania undulata) and three mosses (Amblystegium riparium, Fontinalis antipyretica, Rhynchostegium riparioides). The data to establish these relationships are all based on our own studies, some published already, some here for the first time. They come from a wide range of streams and rivers in Belgium, France, Germany, Ireland, Italy and the U.K. There were significant bivariate positive relationships between concentrations of Zn, Cd and Pb in water and plant for all species except Cd and Pb in Stigeoclonium tenue. When relationships were compared using datasets with total or filtrable metals in water, most differences were slight. However there were marked differences both between species and between metals. Comparison for the seven species of Zn in the plant when aqueous Zn is 0.01 mg l^–1, a concentration at which all seven were found, shows that the four bryophytes had the highest concentrations; however the two green algae had steeper slopes (representing change in concentration in plant in response to change in aqueous concentration). Lemanea fluviatilis had a slope closer to that of the bryophytes, but the concentration was about one order of magnitude lower. All seven species were found at a concentration of 0.01 mg l^–1 Pb, and at this concentration there were almost two orders of magnitude difference between the species which accumulated the most (Scapania undulata) and the one which accumulated the least (Cladophora glomerata). The steepest slope was however shown by C. glomerata.When multiple stepwise regression was applied, the aqueous metal under consideration was the first variable extracted in only nine of the 21 regressions. However one of the other heavy metals (aqueous or accumulated) was extracted first in all but one of the other regressions, presumably because the occurrences of Zn, Cd and Pb were strongly cross-correlated. The principal non-heavy metal factor extracted for Zn and Cd, but not Pb, was aqueous Ca. The relevance of these results to the use of aquatic plants for monitoring heavy metals is discussed.     

重金属Cd、Zn、Cu、Pb对土壤微生物和酶活性的影响

采用室内培养实验(25℃),研究了不同培养时间下重金属Cd、Zn、Cu、Pb(浓度分别为50,800,400,800mg.kg-1)污染对土壤微生物和酶活性的影响。结果表明,土壤蔗糖酶、过氧化氢酶和脱氢酶活性随着培养时间的增加而显著下降,在培养20d的时候达到最小值,然后酶活性缓慢升高。Cu对脲酶活性以及Cd对酸性磷酸酶和脲酶活性的抑制作用随时间增加而增加。土壤微生物生物量碳、细菌、真菌和放线菌数量随培养时间的增加均表现出先降低后升高的变化趋势。Cd和Cu对微生物生物量氮的抑制作用则随着培养时间的增加而增强,在培养30d时微生物生物量氮到达最低值,分别较培养10天减少了12.6%和16.5%。     

Extraction of High Molecular Weight Genomic DNA from Soils and Sediments

The soil microbiome is a vast and relatively unexplored reservoir of genomic diversity and metabolic innovation that is intimately associated with nutrient and energy flow within terrestrial ecosystems. Cultivation-independent environmental genomic, also known as metagenomic, approaches promise unprecedented access to this genetic information with respect to pathway reconstruction and functional screening for high value therapeutic and biomass conversion processes. However, the soil microbiome still remains a challenge largely due to the difficulty in obtaining high molecular weight DNA of sufficient quality for large insert library production. Here we introduce a protocol for extracting high molecular weight, microbial community genomic DNA from soils and sediments. The quality of isolated genomic DNA is ideal for constructing large insert environmental genomic libraries for downstream sequencing and screening applications. The procedure starts with cell lysis. Cell walls and membranes of microbes are lysed by both mechanical (grinding) and chemical forces (β-mercaptoethanol). Genomic DNA is then isolated using extraction buffer, chloroform-isoamyl alcohol and isopropyl alcohol. The buffers employed for the lysis and extraction steps include guanidine isothiocyanate and hexadecyltrimethylammonium bromide (CTAB) to preserve the integrity of the high molecular weight genomic DNA. Depending on your downstream application, the isolated genomic DNA can be further purified using cesium chloride (CsCl) gradient ultracentrifugation, which reduces impurities including humic acids. The first procedure, extraction, takes approximately 8 hours, excluding DNA quantification step. The CsCl gradient ultracentrifugation, is a two days process. During the entire procedure, genomic DNA should be treated gently to prevent shearing, avoid severe vortexing, and repetitive harsh pipetting.Download video file.^{(129M, mp4)}     

Structural study of few p-phenylenediacetate complexes of Mn, Cu, Zn and Cd

Different metal carboxylate complexes are prepared by reactions of manganese (+2) acetate, copper (+2) acetate, zinc (+2) acetate and cadmium (+2) acetate with p-phenylenediacetic acid under ambient condition with or without a nitrogen donor ligands and each of them are characterized by conventional spectroscopic techniques along with crystallography.     

Risk Element (As,Cd, Cu,Pb, and Zn) Contamination of Soils and Edible Vegetables in the Vicinity of Guixi Smelter,South China

Risk element (As, Cd, Cu, Pb, and Zn) contamination in soils and in two edible vegetables (Solanum melongena L. and Capsicum annum L.) was investigated in the vicinity of Guixi Smelter, South China. Soil As concentrations averaged 23.9 mg/kg. Sites near the smelter tailings recorded the highest levels of As and heavy metals in soils. The concentration order of heavy metals in soils was Cd < Pb < Zn < Cu. Cu and Cd in soils were abundant in the exchangeable and bound to carbonate fraction, while Pb and Zn were in the residual fraction, limiting their potential toxicity as pollutants. The proportions of the metals in the mobile fraction followed the order Pb < Zn < Cu < Cd. In Solanum melongena L. and Capsicum annum L., Zn concentration was the highest, followed by Cu, Cd, and Pb, different from that in soils and in the mobile fraction. Concentrations of heavy metals in the labile fractions in soils and in vegetables presented significant correlation (p < 0.05). Both of the two vegetables are not the Cu and Zn accumulators. As for Cd and As, Capsicum annum L. poses a higher risk to animal and human health than Solanum melongena L., with soil-plant transfer coefficients more than three. Root-stem is the main barrier for most of the heavy metals and As in the two vegetables, resulting in higher metal concentrations in roots relative to other plant tissues. The low stem-fruit transfer coefficients for Zn in Solanum melongena L. and for Pb in Capsicum annum L. suggested that very few of them could reach the fruits.     

Effect of Cd,Zn, and Pb Compound Pollution on Celery in a Ferric Acrisol

A pot experiment with an orthogonal experimental design L₉(3⁴) was conducted to study the combined effects of Cd, Zn, and Pb on the growth and metal content of celery grown in a ferric acrisol. The uptake of Cd, Zn, and Pb by celery was not only affected by the individual elements, but also by combinations of the elements. The effect of coexisting elements on plant uptake of the heavy metals depended on the concentration ratios of the elements. There is a given ratio where a maximum antagonism or synergism effect occurs. The combinations of elements clearly affected the dry weight of celery and the heavy metal concentration in celery. The removal rate (the ratio of plant total uptake to the total metal content in soil) was in the order of Cd > Zn > Pb, with no obvious difference between the removal rate under single pollution and that under compound pollution.     

Estimation of Dietary Pb and Cd Intake from Pb and Cd in Blood or Urine

Successful trials were made to estimate the dietary daily intake of lead (Pb) and cadmium (Cd) via foods from the levels of the metals in blood or urine. In practice, 14 and 15 reports were available for Pb and Cd in blood (Pb-B and Cd-B), urine (Pb-U and Cd-U) and 24-h diet duplicates (Pb-D and Cd-D), respectively, from which 68 pairs each of Pb or Cd in blood and food duplicates [each being geometric mean (GM) values for the survey sites] were obtained. Regression analysis revealed that there was a significant correlation between Pb-B and Pb-D, and also between Cd-B and Cd-D, suggesting that it should be possible to estimate both Pb-D and Cd-D from Pb-B and Cd-B, respectively. For Cd-U, the number of available cases was limited (20 pairs), but a significant correlation was detected between Cd-U (as Cd-U_cr, or Cd levels in urine as corrected for creatinine concentration) and Cd-D. Care should be taken in estimating Pb-D from Pb-B, as the ratio of Pb-D over Pb-B may decrease as a function of increasing Pb-B levels. The Pb-D (μg/day) for typical Japanese women with Pb-B of 15 μg/l was best estimated to be 13.5 μg/day. No Cd-B- or Cd-U_cr-dependent change was detected in case of Cd. The best estimate of Cd-D for Cd-B at 1.5 μg/l should be about 19.4 μg/day.     

Geochemical controls for Al, Fe, Mn, Cd, Cu, Pb, and Zn during experimental acidification and recovery of Little Rock Lake, WI, USA

Concentrations of Al, Fe, Mn, Cd, Cu, Pb, and Zn were measured in thereference and treatment basins of Little Rock Lake (Vilas County, Wisconsin), alow-alkalinity, seepage system (pH 6.1, alkalinity25eq/L) during six years of a whole-basinacidificationand the first four years of the lake's recovery. The treatment basin wasacidified with H₂SO₄ in three two-year steps to pH5.6, 5.1, and 4.7. By the end of year 4 of recovery, treatmentbasin pH increased to 5.3 as a result of internal alkalinity generation.During acidification, dissolved Mn and Fe (0.4mpore-size filters) increased at pH 5.6; dissolved Al, Cd, and Zn becameelevated at pH 5.1; and dissolved Pb at pH 4.7. Dissolved Cu remainedsimilar in both basins to pH 4.7. Al, Fe and Mn levels declinedsignificantly during the recovery period, approaching values at pH 5.3intermediate between the concentrations at pH 5.6 and 5.1 during acidification.Dissolved Al and Fe in the reference basin were near the equilibrium levels forsolubility of gibbsite (Al(OH)₃) and amorphousFe(OH)₃(s).The acidified basin was undersaturated relative to gibbsite, and dissolved Alwas limited by pH disequilibrium between the water column and sediments andpossibly by Al-DOC precipitation. Dissolved Fe apparently was controlled bysolubility of amorphous Fe(OH)₃(s) and Fe-DOC precipitation.Dissolved Mn levels in both basins were consistent with manganite[-MnOOH(s)] solubility. Elevated levels of Cd, Pb, and Zn in thetreatment basin during acidification probably resulted from less efficientscavenging of atmospherically-deposited Cd, Pb, and Zn by settling particles.     

Accumulation and Regulation Effects from the Metal Mixture of Zn, Pb, and Cd in the Tropical Shrimp Penaeus vannamei

Environmental metal pollution is one of the major problems faced by humankind. This type of pollution affects aquatic systems (estuaries, coastal lagoons, etc.), which are very dynamic systems, therefore making the study of the effects on the organisms that inhabit them an essential issue. In this study, the capacity of metal regulation by decapod crustacean Penaeus vannamei juveniles was determined. The effects of zinc, lead, and cadmium were tested individually and as a metal mixture exposure to determine possible synergism. The results showed that juvenile shrimps were capable of regulating zinc and lead, whereas cadmium was accumulated without any excretion, at least within the concentrations studied. It was also proved that under the estuarine conditions tested here, P. vannamei juveniles showed capacity to act as a bioindicator for cadmium.     

A comparison of Zn, Mn, Cd, and Ca transport mechanisms in oat root tonoplast vesicles

Oat root tonoplast vesicles were used to determine if tonoplast transport of the divalent cations Zn and Mn occurs via an antiport mechanism, like that described for Ca and Cd. Also, inhibitors reported to affect Ca transport were tested for their effects on Cd versus Ca transport and tonoplast ATPase activity. The ability of Ca, Cd, Zn, and Mn to alter the proton gradient was monitored using both the fluorescent probe acridine orange and C-methylamine accumulation. After the proton gradient was established in MgATP-energized vesicles, addition of Ca, Cd, and Zn to the reaction restored the fluorescence of acridine orange, indicating dissipation of the proton gradient. Fluorescence recovery was linearly correlated with metal concentration and followed the order Ca>Cd≧Zn. Addition of Mn did not restore the fluorescence of acridine orange. All four ions released C-methylamine from MgATP-energized vesicles in an ion-concentration-dependent manner, and with relative initial rates in the order of Ca>Cd>Zn>Mn. The observed ion-concentration-dependent release of protons from sealed vesicles suggests that Zn and Mn, like Ca and Cd, can be antiported into the plant vacuole. In an effort to assess whether Ca and Cd use the same carrier, we tested the effects of verapamil, Do-Tea-Br, nifedipine, ruthenium red, and LaCl₂ on Ca versus Cd transport, and also on MgATPase activity. These compounds are shown to alter Ca transport in plants. Although some of the inhibitors had a negative effect on MgMAPase activity, the decrease in this activity did not account for the decrease in Ca or Cd transport observed in any case. Particularly verapamil had a much greater effect on Ca transport than Cd transport activity while not inhibiting ATPase substantially. Data presented provide evidence for Zn and Mn antiport activity in oat root tonoplast and show differences in responses of Ca and Cd antiport activities to several transport inhibitors.     

Remediation of Pb and Cd Polluted Soils Using In Situ Immobilization and Phytoextraction Techniques

In situ immobilization and phytoextraction techniques have been used for remediation of Pb and Cd polluted soils. Three rates (0.25, 0.5 and 1.0%) of seven immobilizing agents (cement, slag, phosphate rock, bitumen, Fe- and Al-gels, and δ-MnO₂) were tested on three soils containing various levels of Pb (48–192.0 ug/g) and Cd (0.75–3.45 ug/g). All immobilizing agents reduced the plant available Pb and Cd as extracted by DTPA (diethylenetriaminepentaacetic acid). The effectiveness of the various agents in immobilizing Pb and Cd followed the descending order: bitumen > cement > slag > Fe-gel > Al-gel > phosphate rock > δ -MnO₂. Cement and phosphate rock fixed Pb and Cd mainly in the carbonate form, whereas the slag, bitumen, Fe-gel, Al-gel and δ -MnO₂ fixed the metals mainly in the oxide form.

The results of pot experiment proved the high ability of barnyard grass (Echinnochloa stagninum) to accumulate elevated amounts of Pb and Cd (ranging from 291–2421 and 6.1–45.9 ug metal/g dry matter, respectively). These amounts are higher than those reported for hyperaccumulators, particularly for Pb. The amounts of Pb and Cd removed by barnyard grass represent, on average, 46 and 72% of their initial total contents in the soils, respectively. These results proved that, without any other soil treatments, barnyard grass is highly efficient in removing considerable amounts of Pb and Cd from polluted soil within a reasonably short period of time. Therefore, use of barnyard grass for the phytoremediation of Pb and Cd polluted soils is feasible and recommended as an environmentally safe and cheap method. The most significant finding of this study is to name the barnyard grass as an efficient lead accumulator plant.     

Brevibacillus brevis Isolated from Cadmium- or Zinc-Contaminated Soils Improves in Vitro Spore Germination and Growth of Glomus mosseae under High Cd or Zn Concentrations

In this study we investigated the saprophyte growth of two arbuscular–mycorrhizal fungi (Glomus mosseae isolate) under increasing Cd or Zn levels and the influence of a selected bacterial strain of Brevibacillus brevis. Microorganisms here assayed were isolated from Cd or Zn polluted soils. B. brevis increased the presymbiotic growth (germination rate growth and mycelial development) of Glomus mosseae. Spore germination and mycelial development of both G. mosseae isolate were reduced as much as the amount of Cd or Zn increased in the growth medium. In medium supplemented with 20 μg Cd mL⁻¹, the spore germination was only 12% after 20 days of incubation, but the coinoculation with B. brevis increased this value to 40% after only 15 days. The addition of 20 μg Cd mL⁻¹ to the growth medium drastically inhibited hyphal development, but the presence of the bacterium increased hyphal growth of G. mosseae from 195% (without Cd) until 254% (with 20 μg Cd mL⁻¹). The corresponding bacterial effect increasing micelial growth ranged from 125% (without Zn) to 232% (200 μg Zn mL⁻¹) in the case of G. mosseae isolated from Zn-polluted soil. Mycelial growth under 5 μg Cd mL⁻¹ (without bacterium) was similarly reduced from that produced at 15 μg Cd mL⁻¹ in the presence of the bacteria. As well, 50 μg Zn mL⁻¹ (without bacterium) reduced hyphal growth as much as 200 μg Zn mL⁻¹ did in the presence of B. brevis. The bacterial effect on the saprophytic growth of G. mosseae in absence of metal may be due to the involvement of indole acetic acid (IAA) produced by these bacteria. The Cd bioaccumulation ability exhibited (76%) by Cd-adapted B. brevis reduced the Cd damage on G. mosseae in Cd-contaminated medium. These capabilities of B. brevis isolates partially alleviate the inhibitory effects of Cd or Zn on the axenic growth of G. mosseae.