Ecofriendly Heavy Metal Stabilization: Microbial Induced Mineral Precipitation (MIMP) and Biomineralization for Heavy Metals within the Contaminated Soil by Indigenous Bacteria

Heavy metals (HMs) pollution is a serious problem, worldwide. The present study focuses on HMs removal (HMs mobility restricted within soil) by microbial induced mineral precipitation (MIMP) using indigenous isolates. Indigenous isolates were identified as Bacillus subtilis C(225)(MK334652), Oceanobacillus indicireducens 5(225)(MK334653), and Bacillus pumilus 7(225)(MK334654) by 16S-rRNA from HMs polluted industrial-soil (soil-225) that is used for bioremediation. The studied soil contained high level (mg/kg) of Cr(307.41?±?1.02), Cu(576.35?±?1.02), and Zn(708.81?±?1.50) as per Taiwan permissible limit (TPL) and/or Canadian Soil Quality Guidelines (CSQG). Minimum inhibitory concentration (MIC) of Bacillus species were higher compare to Oceanobacillus. Bioavailable-fraction (F₁) of HMs (Cr/Cu/Zn) in untreated 225-soil was noticed to be higher compared to F₂–F₅ fraction. Bacillus subtilis C(225) are more efficient for HMs-mineralization, compared to other isolates where Cr/Cu/Zn immobilized within soil, transforming from bioavailable-fraction (F₁) (high-mobility) (60–90% reduction) to residual-fraction F₅ (low-mobility) (50–80% increment). Organic-bound-fraction (F₄) (medium-mobility) of Cr increased (42.66?±?2.1%), compared to Cu/Zn, by Bacillus subtilis C(225). Urease (40.8%) and dehydrogenase (8.3%) activity increased in bio-remediated soil, compared to control. MIMP of HMs by ingenious bacteria reflects the transformation-pathway from F₁ to F₅ of HMs to fix strongly within soils with inadequate mobility. Thus, MIMP by indigenous microbe can be applicable for HMs stabilization/removal in contaminated soil for ecofriendly environmental cleanup.     

Effects of arbuscular mycorrhizal fungi on the growth and heavy metal accumulation of bermudagrass [Cynodon dactylon (L.) Pers.] grown in a lead–zinc mine wasteland

Abstract

A pot experiment was conducted to investigate the potential influence of arbuscular mycorrhizal fungi (AMF), Funneliformis mosseae and Diversispora spurcum, on the growth and nutrient (P and S) and heavy metal (HMs) (Pb, Zn, and Cd) content of bermudagrass [Cynodon dactylon (L.) Pers.] in a lead-zinc mine wasteland. The D. spurcum inoculation significantly increased the bermudagrass growth, whereas the F. mosseae inoculation did not. The AMF inoculation significantly increased the soil pH and uptake of P, S, and HMs by bermudagrass, decreased the contents of available Pb and Zn in soils and Pb in shoots, reduced the translocation factor (TF) and translocation capacity factor (TF') of Pb and Cd in bermudagrass and increased the TF and TF' of Zn in bermudagrass. A significant negative correlation was found between pH and available HMs in soil, whereas a significant positive correlation was noted between the HMs content and nutrient content in bermudagrass shoots. Experiment results provide evidence of the potential role of AMF in improving bermudagrass performance for the vegetation restoration of metalliferous mine wastelands.     

Geochemical speciation and ecological risk assessment of heavy metals in surface soils collected from the Yellow River Delta National Nature Reserve,China

In this study, a comprehensive assessment of soil heavy metal (HMs) pollution in the Yellow River Delta National Nature Reserve (YRDNNR) was conducted. Spatial distributions, chemical fractions, and sources of eight HMs (Cu, Zn, Pb, Cr, Cd, Fe, Mn, and Ni) in 46 soil samples in the studied region were analyzed. In addition, the potential risks of the HMs were evaluated. The results showed that the mean concentrations of Cu, Zn, Pb, Cr, Cd, Fe, Mn, and Ni were 19.4, 65.2, 38.4, 55.9, 0.078, 41546.5, 510.3, and 27.5 mg kg^?1, respectively. It indicates that the concentrations of most HMs, with exception of Pb and Fe, in samples were similar to the background value of soil in China. Principal component analysis results showed that the HMs originated mainly from natural sources, but Pb pollution in the studied area was significantly caused by anthropogenic activities. In addition, Ecological risk assessment statistical analysis indicates that the HM contamination level in YRDNNR ranged from low to moderately polluted, however, the environmental risk due to Mn and Pb contamination was high.     

Assessment of Heavy Metals Contamination and Potential Ecological Risk in Soils Affected by a Former Mn Mining Activity,Drama District,Northern Greece

Manganese mining activities in the Drama district, northern Greece, have resulted in a legacy of abandoned mine wastes at the “25 km Mn-mine” site. Current research was focused on the western Drama plain (WDP), constituting the recipient of the effluents from Xiropotamos stream, which passes through the “25 km Mn-mine” place. A total of 148 top soil samples were collected and their heavy metals (HMs) concentrations (Mn, Pb, Zn, Cu, Cd, and As) were determined using inductively coupled plasma mass spectrometry. Enrichment factor (EF), geoaccumulation index (Igeo), and pollution load index (PLI) were calculated as an effort to assess metal accumulation, distribution, and pollution status of the soils due to the former mining activity. The overall potential ecological risk of HMs to the environment was also evaluated using the potential toxicity response index (RI). Results showed that peak values of the elements (13 wt% for Mn, 0.2 wt% for Pb, 0.2 wt% for Zn, 0.1 wt% for As, 153 mg/kg for Cu, and 27.5 mg/kg for Cd) were found in soils from sites close to and along both sides of the Xiropotamos stream. In this sector of WDP, values of EF, Igeo, and PLI classify the soils as moderately to highly polluted with Mn, Pb, Zn, Cd, and As. Based on RI values, soils in this part of WDP display considerable to very great potential ecological risk and, therefore, a remediation has to be applied. The main cause of soil contamination is considered the Xiropotamos downstream transfer and dispersion of Mn mine wastes via flooding episodes.     

Heavy metal contamination of soil and tree-ring in urban forest around highway in Shanghai,China

The heavy metal (HM) pollution of forest soil has been known as one of the most challenging pollution issues due to their characteristics. In order to know the HM pollution condition in urban forest, identify the possible source of HM, 102 sub-samples of soil in 34 sites and 39 tree rings sub-samples in 7 sites were collected in the outer-ring greenbelt (ORG) in Shanghai, China. Concentrations of Cu, Zn, Pb, and Cd in soil and tree rings were analyzed, and the soil properties pH, total nitrogen, total phosphorous, and organic matter were analyzed too. Geo-accumulation index and potential ecological risk index were used for assessing the contamination level of HMs. Nonparametric tests, one-way analysis of variance, correlation analysis, and principal component analysis were applied. The results showed that: (1) concentrations of Cu, Zn, and Cd in soil were significantly higher than their corresponding background values of Shanghai (BVs); concentrations of Cu, Zn, and Pb in tree rings increased gradually in the past 10 years; (2) Zn and Cd were in unpolluted to moderately polluted level, Cd has moderate degree potential ecological risk; (3) vehicle exhausts and abrasion of vehicle parts of tires and historical agricultural activities were the main sources of HM contamination; (4) Cinnamomum camphora (L.) Presl. has the potential to reconstruct the change of Cu, Zn, and Pb as a bioindicator. In conclusion, Cd should be considered as a priority control component. The relationship between plant and soil should take further focus and more studies of the behavior of HMs in soil and plants are required.     

Early Toxic Effects of Zinc, Cobalt, and Cadmium on Photosynthetic Activity of the Green AlgaChlorella pyrenoidosaChick S-39

Early toxic effects of heavy metals (HMs) Zn, Co, and Cd at concentrations from 0.01 to 100 mM on photosynthetic activity of the green algaChlorella pyrenoidosaChick S-39 were studied. The early effect of HMs was manifested as a rapid (within 0.5–2 h) reduction of photoinduced oxygen release by the algal cells. The suppressed relative yield of variable chlorophyll fluorescence (F _V /F _m ) by HMs as well as its dynamics inC. pyrenoidosaprovided evidence for rapid inactivation of photosystem II (PS II). Analysis of the induction curve of delayed chlorophyll fluorescence inChlorellacells suggested that the early toxic effects of Zn, Co, and Cd at the above concentrations manifested itself not only in inhibited electron transport in PS II, but also in reduced energization of photosynthetic membranes. Hence, the early toxic effect of Zn, Co, and Cd was primarily related to the decreased efficiency of the light reactions of photosynthesis, which subsequently resulted in reduced productivity of the alga.     

Integrative Application of Soil Conditioners and Bio-augmentation for Enhanced Heavy Metal Stabilization from Wastewater and Improved Growth of Nicotiana alata L. and Petunia hydrida L.

Wastewater generated from industries contains numerous contaminants, among which heavy metals (HMs) are non-degradable. This research work highlights the use of commonly used ornamental plants, Nicotiana alata L. and Petunia hydrida L., with compost (C) and peat moss (M), and rhizospheric bacterial augmentation using Pseudomonas japonica, for the phytostabilization of HMs from synthetic wastewater. After plant–soil acclimatization, plants were exposed for 6 weeks to synthetic wastewater, containing cadmium, chromium, copper, lead, nickel, and zinc concentrations (based on the HMs level of wastewaters collected from textile and pharmaceutical industry). Physiological response, biochemical status, and enzymatic fluctuations of plants and the distribution of HMs in plant parts and soil, were quantified. With the combined use (5% each v/v conditioner/soil) of C and M, in bio-augmented soil, physiological response and enzymatic status of both plants improved, with decreased stress injury due to HMs. Further, the plant HMs uptake was reduced, with better stabilization of HM in soil. For better phytostabilization of HMs in wastewater, the use of compost, peat moss, and bacterial augmentation is recommended with Nicotiana alata L. and Petunia hydrida L.

     

Health Risk Assessment of Consumption of Heavy Metals in Market Food Crops from Sialkot and Gujranwala Districts,Pakistan

This study was performed to investigate the potential health risk of heavy metals (HMs) through consumption of market food crops (MFCs) in the Sialkot and Gujranwala districts, Pakistan. Both study areas are located in industrialized regions of the country, where atmospheric pollution is a problem and irrigation of food crops is mostly practiced on the use of wastewater/contaminated water. For the purpose of this study, MFCs samples were collected and assessed for HMs (Cr, Ni, Cd, Pb, Mn, Cu, Zn, and Fe) by using flame atomic absorption spectrophotometry. Concentration of HMs such as Pb and Cd exceeded the Food and Agriculture/World Health Organization's recommended limits in all MFCs, while Cr in most of the vegetables of the Sialkot and Gujranwala districts also exceeded that limit. The health risk index was >1 in Triticum aestivum for Pb and Cd intake in the Sialkot district and only Pb in the Gujranwala district. Therefore, this study suggests pretreatment of wastewater and its utilization for lawns and green belts irrigation, rather than for food crops. This study also suggests a regular monitoring of HMs in the irrigation water, subsequent soil, air, and MFCs in order to prevent or reduce health hazards.     

Effects of Long-Term exposure to Heavy Metals upon Rhizosphere Bacteria from Baia Mare Area (Maramureş County,Romania)

Abstract

The aim of this study was to investigate the extent of heavy metal (HM) pollution and its effect on microorganisms from rhizosphere soil in Baia Mare area (Maramure? County, Romania). Two sites with different contamination degrees were included in the study: one with a long history of mining activities and one within a drinking water safeguard zone. Rhizosphere soil samples were characterized with respect to physico-chemical parameters and the Cd, Cu, Pb and Zn contents. Native bacteria were investigated for HM tolerance and biofilm formation under toxic exposure by the microdilution assay. The most resistant strains were identified and the minimum inhibitory concentrations for HMs were determined. Cd, Cu, Pb and Zn exceeded the intervention threshold in Bozânta tailings site, while Pb content exceeded the intervention level within the area of the drinking water treatment plant. Cd showed a very high potential ecological risk in Bozânta area. The long-term exposure to HMs contributed to the selection of HM-tolerant and weakly adherent strains. Biofouling was significantly reduced under the influence of copper ions. Arthrobacter, Rhodococcus and Acidovorax strains with exceptional resistant profiles were isolated from the tailings site, indicating the important role of native microorganisms in rhizosphere ecosystems of contaminated sites.     

Thermal Characteristics of Hyperaccumulator and Fate of Heavy Metals during Thermal Treatment of Sedum plumbizincicola

Thermal treatment is one of the most promising disposal techniques for heavy metal- (HM)-enriched hyperaccumulators. However, the thermal characteristics and fate of HMs during thermal treatment of hyperaccumulator biomass need to be known in detail. A horizontal tube furnace was used to analyze the disposal process of hyperaccumulator biomass derived from a phyto-extracted field in which the soil was moderately contaminated with heavy metals. Different operational conditions regarding temperature and gas composition were tested. A thermo-dynamic analysis by advanced system for process engineering was performed to predict HM speciation during thermal disposal and SEM-EDS, XRD and sequential chemical extraction were used to characterize the heavy metals. The recovery of Zn, Pb and Cd in bottom ash decreased with increasing temperature but recovery increased in the fly ash. Recovery of Zn, Pb and Cd fluctuated with increasing air flow rate and the metal recovery rates were higher in the fly ash than the bottom ash. Most Cl, S, Fe, Al and SiO₂ were found as alkali oxides, SO₂, Fe₂(SO₄)₃, iron oxide, Ca₃Al₂O₆, K₂SiO₃ and SiO₂ instead of reacting with HMs. Thus, the HMs were found to occur as the pure metals and their oxides during the combustion process and as the sulfides during the reducing process.     

Factors affecting immobilization of heavy metals by purple nonsulfur bacteria isolated from contaminated shrimp ponds

In order to remove heavy metals (HMs) from contaminated shrimp pond at the highest concentrations found of; 0.75 mg/l Cd²⁺, 62.63 mg/l Pb²⁺, 34.60 mg/l Cu²⁺ and 58.50 mg/l Zn²⁺, two strains of purple nonsulfur bacteria isolated from shrimp ponds (NW16 and KMS24) were investigated for their ability to immobilize HMs in 3% NaCl in both microaerobic-light and aerobic-dark conditions. Based on metabolic inhibition and metabolic-dependent studies, it was concluded that both strains removed HMs using biosorption and also bioaccumulation. The efficiency of removal by both strains with both incubating conditions tested was in the order of lead (Pb) > copper (Cu) > zinc (Zn) > cadmium (Cd). Optimal conditions for removal of HMs by strain NW16 were; cells in the log phase at 4.5 mg DCW/ml, pH 6.0, and 30°C for 30 min. With microaerobic-light conditions, the relative percent removal of HMs was: Pb, 83; Cu, 59; Zn, 39; Cd, 23 and slightly more with the aerobic-dark conditions (Pb, 90; Cu, 69; Zn, 46; Cd, 28). Cells in the log phase at 5.0 mg DCW/ml, pH 5.5, and 35°C for 45 min were optimal conditions for strain KMS24 and there were no significant differences for the removal percentages of HMs with either incubating conditions (averages: Pb, 96; Cu, 75; Zn, 46; Cd, 30). The presence of Ca²⁺ and Mg²⁺ significantly decreased the removal capacity of HMs for both strains.     

Enhancement of toxic Cr (VI), Fe,and other heavy metals phytoremediation by the synergistic combination of native Bacillus cereus strain and Vetiveria zizanioides L

Bioremediation of Cr (VI), Fe, and other heavy metals (HMs) through plant–microbes interaction is one of the efficient strategies due to its high efficiency, low cost, and ecofriendly nature. The aim of the study was to isolate, characterize, and assess the potential of rhizospheric bacteria to enhance growth and metal accumulation by the chromium hyperaccumulator Vetiveria zizanoides. The bacterial strain isolated from mine tailings was identified to be Bacillus cereus (T1B3) strain exhibited plant growth-promoting traits including, 1-aminocyclopropane-1-carboxylate deaminase, indole acetic acid, and siderophores production, nitrogen fixation, and P solubilization. Removal capacity (mg L^?1) of T1B3 strain was 82% for Cr⁺⁶ (100), 92% for Fe (100), 67% for Mn(50), 36% for Zn (50), 31% for Cd (30), 25% for Cu (30), and 43% for Ni (50) during the active growth cycle in HM-amended, extract medium. Results indicate that inoculating the native V. zizanioides with T1B3 strain improves its phytoremediation efficiency of HMs. The mineralogical characteristics of chromite ore tailings and soil were also confirmed by X-ray diffraction, Fourier Transform Infrared, scanning electron microscope–energy dispersive spectroscopy analysis.     

Spatial distribution and human health risk assessment of heavy metals in campus dust: A case study of the university town of Huaxi

Abstract

A detailed investigation was conducted to understand the contamination characteristics of a selected set of heavy metals (HMs) in 34 campus dust samples from Huaxi University Town. The HMs spatial distribution analysis based on ArcGIS software, the geo-accumulation index (I_geo) and health risk model were employed for evaluation, and multivariate statistical methods were used to identify possible sources. Results showed that the mean concentrations of Cu, Zn, Pb, Cr, Ni, and Cd were 68.18, 123.81, 45.26, 140.36, 47.26, and 0.47?mg/kg, respectively. The spatial distribution characteristics displayed that the relatively large concentrations for the analyzed HMs were mainly located at both teaching areas and students’ dormitory areas. The average values of I_geo indicated that HMs contamination level followed the decreasing trend of Cd?>?Cu?>?Cr?>?Pb?>?Ni?>?Zn. The health risk assessment results indicated that HMs in campus dust generally do not pose any immediate health risk for both adult males and adult females but the cumulative effect is a matter of concern. The sources analyses demonstrated that Cu, Zn, Cr, and Cd, primarily from motor vehicle emission and waste incineration, Pb predominantly originated from construction source, while Ni had the mixed sources of nature and traffic.     

Effects of Cadmium,Copper, Lead,and Zinc Contamination on Metal Accumulation by Safflower and Wheat

Accumulation of heavy metals (HMs) in cultivated soils is a continuing environmental problem in many parts of the world. An increase in HM concentration can enhance uptake of toxic metals by crops and enter the human food chain. In this study, the uptake behavior of wheat and safflower was evaluated in a calcareous soil by using 12 undisturbed columns in which half were artificially contaminated. Heavy metals in the form of CdCl₂ (15 mg Cd kg^{? 1}), CuSO₄ (585 mg Cu kg^{? 1}), Pb(NO₃)₂ (117 mg Pb kg^{? 1}), and ZnCl₂ (1094 mg Zn kg^{? 1}) were sprayed on the soil surface and completely mixed in the top 10 cm. The background total concentrations of Cd, Cu, Pb and Zn were 1.6, 29.5, 17.5 and 61.2 mg kg^{? 1}, respectively. After metal application, half of the columns (3 contaminated and 3 uncontaminated) were sown with wheat (Triticum aestivum) and the other half with safflower (Carthamus tinctorious) and grown for 74 days until maturity. After harvesting, soil columns were cut into 10-cm sections and analyzed for HNO₃- and DTPA-extractable metal concentrations. Metal concentrations were also measured in different plant tissues. The results showed that artificial contamination of topsoil decreased the transpiration rate of wheat by 12% and that of safflower by 6%. In contaminated columns, Cd, Cu, Pb, and Zn accumulation in wheat shoot was greater by 8.0-, 1.9-, 3.0-, and 2.1-fold than the control, respectively. Accordingly, these numbers were 46.0-, 1.3-, 1.7-, and 1.6-fold in safflower shoot. Soil contamination with HMs resulted in a 55% decrease in shoot dry matter yield of wheat while it had no significant effect on shoot dry matter of safflower. The normalized water consumption for safflower was therefore not affected by metal contamination (≈ 13 mm H₂O g^{? 1} of dry weight for all safflower and uncontaminated wheat treatments), while contaminated wheat was much less water efficient at about 27 mm H₂O g^{? 1} dry weight. It was concluded that although artificial contamination had a negative effect on wheat growth, it did not affect safflower's normal growth and water efficiency.     

Long-term field phytoextraction of zinc/cadmium contaminated soil by Sedum plumbizincicola under different agronomic strategies

In two long-term field experiments the zinc (Zn)/cadmium (Cd) hyperaccumulator Sedum plumbizincicola (S. plumbizincicola) was examined to optimize the phytoextraction of metal contaminated soil by two agronomic strategies of intercropping with maize (Zea mays) and plant densities. Soil total Zn and Cd concentrations decreased markedly after long-term phytoextraction. But shoot biomass and Cd and Zn concentrations showed no significant difference with increasing remediation time. In the intercropping experiment the phytoremediation efficiency in the treatment “S. plumbizincicola intercropped with maize” was higher than in S. plumbizincicola monocropping, and Cd concentrations of corn were below the maximum national limit. In the plant density experiment the phytoremediation efficiency increased with increasing plant density and 440,000 plants ha^?1 gave the maximum rate. These results indicated that S. plumbizincicola at an appropriate planting density and intercropped with maize can achieve high remediation efficiency to contaminated soil without affecting the cereal crop productivity. This cropping system combines adequate agricultural production with soil heavy metal phytoextraction.     

Leaching and microbial treatment of a soil contaminated by sulphide ore ashes and aromatic hydrocarbons

Contaminated soil from a historical industrial site and containing sulfide ore ashes and aromatic hydrocarbons underwent sequential leaching by 0.5 M citrate and microbial treatments. Heavy metals leaching was with the following efficiency scale: Cu (58.7%) > Pb (55.1%) > Zn (44.5%) > Cd (42.9%) > Cr (26.4%) > Ni (17.7%) > Co (14.0%) > As (12.4%) > Fe (5.3%) > Hg (1.1%) and was accompanied by concomitant removal of organic contaminants (about 13%). Leached metals were concentrated into an iron gel, produced during ferric citrate fermentation by the metal-resistant strain BAS-10 of Klebsiella oxytoca. Concomitantly, the acidic leached soil was bioaugmented with Allescheriella sp. DABAC 1, Stachybotrys sp. DABAC 3, Phlebia sp. DABAC 9, Pleurotus pulmonarius CBS 664.97, and Botryosphaeria rhodina DABAC P82. B. rhodina was most effective, leading to a significant depletion of the most abundant contaminants, including 7-H-benz[DE]anthracene-7-one, 9,10-anthracene dione and dichloroaniline isomers, and to a marked detoxification as assessed by the mortality test with the Collembola Folsomia candida Willem. The overall degradation activities of B. rhodina and P. pulmonarius appeared to be significantly enhanced by the preliminary metal removal.     

Bioaccumulation and translocation of nine heavy metals by Eichhornia crassipes in Nile Delta,Egypt: perspectives for phytoremediation

Abstract

The current research was carried out to estimate the potential of water hyacinth (WH) for removal of nine heavy metals (HMs) from three irrigation canals in Nile Delta. Sampling was achieved in monospecific and homogeneous WH stands at three irrigation canals in the study area, and WH biomass was sampled at monthly intervals from April 2014 to November 2014 using five randomly distributed quadrats (each 0.5?×?0.5?m) at each canal. All HM concentrations were significantly higher in the roots compared with the other WH organs. The WH was recognized by a bioaccumulation factor >1.0 for all HMs. The WH was recognized by translocation factor <1.0 for all HMs (except Pb). In many cases, the concentrations of the HMs in the different organs of WH were correlated with the same HMs in the water. Such correlations indicate that WH reflects the cumulative influences of environmental pollution from the water, and thereby suggesting its potential use in the bio-monitoring of most examined HMs. In conclusion, WH is a promising macrophyte for remediation of irrigation canals polluted with Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, and Zn.     

Heavy metal uptake by arbuscular mycorrhizas of Elsholtzia splendens and the potential for phytoremediation of contaminated soil

A pot culture experiment and a field experiment were carried out separately to study heavy metal (HM) uptake from soil contaminated with Cu, Zn, Pb and Cd by Elsholtzia splendens Nakai ex F. Maekawa inoculated with arbuscular mycorrhizal (AM) fungi and the potential for phytoremediation. The HM-contaminated soil in the pot experiment was collected from the field experiment site. Two AM fungal inocula, MI containing only one AM fungal strain, Glomus caledonium 90036, and M II consisting of Gigaspora margarita ZJ37, Gigaspora decipens ZJ38, Scutellospora gilmori ZJ39, Acaulospora spp. andGlomus spp., were applied to the soil under unsterilized conditions. In the pot experiment, the plants were harvested after 24 weeks of growth. Mycorrhizal colonization rate, plant dry weight (DW) and P, Cu, Zn, Pb, Cd concentrations were determined. MI-treated plants had higher mycorrhizal colonization rates than MII-treated plants. Both MI and MII increased shoot and root DW, and MII was more effective than MI. In shoots, the highest P, Cu, Zn and Pb concentrations were all observed in the plants treated with MII, while MI decreased Zn and Pb concentrations and increased P but did not alter Cu, and Cd concentrations were not affected by either of two inocula. In roots, MII increased P, Zn, Pb concentrations but did not alter Cu and Cd, and MI did not affect P, Cu, Zn, Pb, Cd concentrations. Cu, Zn, Pb, Cd uptake into shoots and roots all increased in MII-treated plants, while in MI-treated plants, Cu and Zn uptake into shoots and Cu, Zn, Pb, Cd into roots increased but Pb and Cd uptake into shoots decreased. In general, MII was more effective than MI in promoting plant growth and HM uptake. The field experiment following the pot experiment was carried out to investigate the effects of MII under field conditions. The 45-day-old nonmycorrhizal and MII-colonized seedlings of E. splendens were transplanted to HM-contaminated plots and harvested after 5 months. MII-inoculation increased shoot DW and shoot P, Cu, Zn, Pb concentrations significantly but did not alter shoot Cd concentrations, which led to higher uptake of Cu, Zn, Pb, Cd by E. splendens shoots. These results indicate that the AM fungal consortium represented by MII can benefit phytoextraction of HMs and therefore play a role in phytoremediation of HM-contaminated soils.     

Distribution of Zn in functionally different leaf epidermal cells of the hyperaccumulator Thlaspi caerulescens

The aim of this study was to show the potential of Thlaspi caerulescens in the cleaning‐up of a moderately Zn ‐contaminated soil and to elucidate tolerance mechanisms at the cellular and subcellular level for the detoxification of the accumulated metal within the leaf. Measured Zn concentrations in shoots were high and reached a maximum value of 83 mmol kg ^{? 1} dry mass, whereas total concentrations of Zn in the roots were lower (up to 13 mmol kg ^{? 1}). In order to visualize and quantify Zn at the subcellular level in roots and leaves, ultrathin cryosections were analysed using energy‐dispersive X‐ray micro‐analysis. Elemental maps of ultrathin cryosections showed that T. caerulescens mainly accumulated Zn in the vacuoles of epidermal leaf cells and Zn was almost absent from the vacuoles of the cells from the stomatal complex, thereby protecting the guard and subsidiary cells from high Zn concentrations. Observed patterns of Zn distribution between the functionally different epidermal cells were the same in both the upper and lower epidermis, and were independent of the total Zn content of the plant. Zinc stored in vacuoles was evenly distributed and no Zn‐containing crystals or deposits were observed. From the elemental maps there was no indication that P, S or Cl was associated with the high Zn concentrations in the vacuoles. In addition, Zn also accumulated in high concentrations in both the cell walls of epidermal cells and in the mesophyll cells, indicating that apoplastic compartmentation is another important mechanism involved in zinc tolerance in the leaves of T. caerulescens.     

Combining chemical flocculation and bacterial co-culture of Cupriavidus taiwanensis and Ureibacillus thermosphaericus to detoxify a hardwood hemicelluloses hydrolysate and enable acetone–butanol–ethanol fermentation leading to butanol

Butanol, a fuel with better characteristics than ethanol, can be produced via acetone–butanol–ethanol (ABE) fermentation using lignocellulosic biomass as a carbon source. However, many inhibitors present in the hydrolysate limit the yield of the fermentation process. In this work, a detoxification technology combining flocculation and biodetoxification within a bacterial co-culture composed of Ureibacillus thermosphaericus and Cupriavidus taiwanensis is presented for the first time. Co-culture-based strategies to detoxify filtered and unfiltered hydrolysates have been investigated. The best results of detoxification were obtained for a two-step approach combining flocculation to biodetoxification. This sequential process led to a final phenolic compounds concentration of 1.4 g/L, a value close to the minimum inhibitory level observed for flocculated hydrolysate (1.1 g/L). The generated hydrolysate was then fermented with Clostridium acetobutylicum ATCC 824 for 120 h. A final butanol production of 8 g/L was obtained, although the detoxified hydrolysate was diluted to reach 0.3 g/L of phenolics to ensure noninhibitory conditions. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2753, 2019.