A New Fungal Isolate,Penidiella sp. Strain T9, Accumulates the Rare Earth Element Dysprosium

With an aim to develop a highly efficient method for the recovery of rare earth elements (REEs) by using microorganisms, we attempted to isolate dysprosium (Dy)-accumulating microorganisms that grow under acidic conditions from environmental samples containing high concentrations of heavy metals. One acidophilic strain, T9, which was isolated from an abandoned mine, decreased the concentration of Dy in medium that contained 100 mg/liter Dy to 53 mg/liter Dy after 3 days of cultivation at pH 2.5. The Dy content in the cell pellet of the T9 strain was 910 μg/mg of dry cells. The T9 strain also accumulated other REEs. Based on the results of 28S-D1/D2 rRNA gene sequencing and morphological characterization, we designated this fungal strain Penidiella sp. T9. Bioaccumulation of Dy was observed on the cell surface of the T9 strain by elemental mapping using scanning electron microscopy-energy dispersive X-ray spectroscopy. Our results indicate that Penidiella sp. T9 has the potential to recover REEs such as Dy from mine drainage and industrial liquid waste under acidic conditions.     

Prokaryotic and eukaryotic community structure affected by the presence of an acid mine drainage from an abandoned gold mine

The acid mine drainage that originates in the abandoned gold mine in San Luis, Argentina, is released into La Carolina stream. The aim of this study was to determine the influence of this mine drainage on the physicochemical parameters of the area studied and on both prokaryotic and eukaryotic community structure. In addition, specific relationships between microbial taxonomic groups and physicochemical parameters were established. The drainage that flows into La Carolina stream acidifies the stream and increases its sulfate, Zn, Cd and Te concentrations. Microbial analysis showed that prokaryotic community structure is mainly affected by pH values. Actinobacteria and Gammaproteobacteria were abundant in samples characterized by low pH values, while Nitrospirae, Chloroflexi, Deltaproteobacteria, Thaumarchaeota and Euryarchaeota were associated with high concentrations of heavy metals. Otherwise, Alphaproteobacteria was present in samples taken in sunlit areas. Regarding eukaryotic community structure, the sunlight had the greatest impact. Inside the mine, in the absence of light, fungi and protists members were the most abundant microorganisms, while those samples taken in the presence of light displayed algae (green algae and diatoms) as the most abundant ones. After receiving the mine drainage, the stream showed a decrease in the diatom abundance and green algae predominated.

     

Effect of high concentration of Co (II) on Enterobacter liquefaciens strain C-1: a bacterium highly resistant to heavy metals with an unknown genome

Heavy metals are required as nutrients for essential functions in microorganisms. However, higher concentrations of these cations are generally toxic and may produce contrasting effects on living organisms. Enterobacter liquefaciens strain C-1, a bacterium isolated from the Moa mine in Cuba, is able to survive in the presence of high concentrations of heavy metals. The proteomes of Enterobacter liquefaciens strain C-1, grown under aerobic conditions in the presence and absence of Co (II) were compared using two-dimensional gel electrophoresis analysis in the isoelectric point range of 4-7 and the mass range of 15-120 kDa. Significant changes in the expression level (> two-fold) were detected for 13 spots: seven and six were up- and down-regulated, respectively. Because the genome of this bacterium is unknown, identification by peptide mass fingerprinting only succeeded in four cases and most of the cross-species identifications were supported by de novo sequencing of tryptic peptides followed by sequence alignment using the MS BLAST program. Twelve different proteins were identified, ten are involved in cellular antioxidant defence probably induced by the presence of Co (II). This is the first step towards understanding the role of proteins participating in the mechanism of resistance to heavy metals in this bacterium.     

矿井废水灌溉对小麦生理特性及重金属积累的影响

以井水灌溉为对照 (CK),采用盆栽试验研究了矿井废水灌溉对小麦生理特性和重金属积累的影响.设置了3个矿区废水灌溉处理:洗煤废水(T₁)、经沉淀处理的洗煤废水 (T₂) 和煤矸石淋溶水 (T₃),于返青期开始进行矿区废水灌溉处理.结果表明： 矿井废水灌溉处理对小麦的生长发育和产量均有不同程度的负面影响.到开花期时,T₁、T₂和T₃处理小麦的单茎质量和叶面积、根系活力和光合速率均显著低于对照(P＜0.05),T₃处理小麦株高和叶绿素含量(SPAD值)显著降低(P＜0.05);T₁、T₂和T₃处理的籽粒产量分别比对照下降15.4%、9.8%和17.8%.此外,矿井废水灌溉处理小麦籽粒中Cr、Pb、Cu和Zn的含量均显著高于对照,表明矿井废水灌溉导致重金属在小麦籽粒中积累.
     

微生物与重金属间的相互作用及其应用研究

从多方面阐述了微生物与重金属二者间相互作用,指出微生物在生长代谢过程中能淋滤、吸收和转化重金属,对重金属有一定的抗性和解毒作用;但是,一定浓度的重金属对微生物过程及其种群具有较大的毒性。影响微生物在环境介质中的活动,矿业工程生产工艺已充分利用微生物能淋滤,吸收和转化重金属等特性来处理低品位难浸矿石,环境保护领域也积极利用微生物对重金属的抗性和解毒作用来实现工业废弃物的处理以及被重金属污染土壤的修复。利用微生物的生物量及其活性可以评价环境中不同介质的重金属污染水平。     

Role of extracellular polymeric substance (EPS) in toxicity response of soil bacteria Bacillus sp. S3 to multiple heavy metals

Heavy metal resistant bacteria are of great interest because of their potential use in bioremediation. Understanding the survival and adaptive strategies of these bacteria under heavy metal stress is important for better utilization of these bacteria in remediation. The objective of this study was to investigate the role of bacterial extracellular polymeric substance (EPS) in detoxifying against different heavy metals in Bacillus sp. S3, a new hyper antimony-oxidizing bacterium previously isolated from contaminated mine soils. The results showed that Bacillus sp. S3 is a multi-metal resistant bacterial strain, especially to Sb(III), Cu(II) and Cr(VI). Toxic Cd(II), Cr(VI) and Cu(II) could stimulate the secretion of EPS in Bacillus sp. S3, significantly enhancing the adsorption and detoxification capacity of heavy metals. Both Fourier transform infrared spectroscopy (FTIR) and three-dimensional excitation–emission matrix (3D-EEM) analysis further confirmed that proteins were the main compounds of EPS for metal binding. In contrast, the EPS production was not induced under Sb(III) stress. Furthermore, the TEM–EDX micrograph showed that Bacillus sp. S3 strain preferentially transported the Sb(III) to the inside of the cell rather than adsorbed it on the extracellular surface, indicating intracellular detoxification rather than extracellular EPS precipitation played an important role in microbial resistance towards Sb(III). Together, our study suggests that the toxicity response of EPS to heavy metals is associated with difference in EPS properties, metal types and corresponding environmental conditions, which is likely to contribute to microbial-mediated remediation.

     

Molecular Characterization of Endophytic Bacteria from Metal Hyperaccumulator Aquatic Plant (Eichhornia crassipes) and Its Role in Heavy Metal Removal

In this study, among a collection of heavy metals resistant endophytic bacterial strains isolated from aquatic hyperaccumulator plant (Eichhornia crassipes), one plant growth promoting endophytic bacteria (PGPE), SVUB4 was selected for its ability to utilize 1-aminocyclopropane-1-carboxylic acid (ACC) as the sole N source and accumulate different heavy metals. The SVUB4 strain was characterized as Enterobacter sp. on the basis of its 16S rDNA sequences. Assessment of the parameters of plant growth promotion revealed the intrinsic ability of the strain for the production of IAA, siderophore and solubilization of insoluble phosphate. Furthermore, plasmid DNA analysis of Enterobacter sp. strain SVUB4 indicated the presence of a single large plasmid element. The results of plasmid curing experiments demonstrated that the ability of this strain to grow in presence of Cd and Zn was encoded by the 98 kb plasmid, whereas the ability to grow in the presence of Pb appeared to be encoded by the chromosome. The Cd and Zn removal capacity of the respective metal sensitive strain (plasmidless) were about 36 and 45 μg/g-1 DW, respectively, while the removal capacity of the both metal by metal resistant strain (p SVUB4) showed a significantly higher Cd and Zn removal capacity of 153 and 228 μg/g^?1 DW, respectively. However, both strains exhibited a similar pattern of Pb accumulation. The present observation also showed that for wild-type strain SVUB4 (pSVUB4), the overall level of IAA production in the absence and in the presence of Cd²⁺ or Zn²⁺was approximately the same. Nevertheless, strain SVUB4M in this respect appeared to be more sensitive to heavy metals: a noticeable decrease in IAA production was observed under the effect of both metals, especially with Cd²⁺.     

Aerobic bioreduction of nickel(II) to elemental nickel with concomitant biomineralization

Although microorganisms have the potential to reduce metals, products with elementary forms are unusual. In the present study, a strain of Pseudomonas sp. MBR was tested for its ability to reduce metal ions to their elementary forms coupled to biomineralization under aerobic conditions. The Pseudomonas sp. MBR strain was able to reduce metals such as Fe(III), Mn(II), Cu(II), Ni(II), Cd(II), Co(II), Al(III), Se(IV), and Te(IV) as electron acceptors to elementary forms using citrate, lactate, pyruvate, succinate, malate, glucose, or ethanol as electron donors. Growth and reduction during biomineralization occurred within the pH range of 6.0 to 11.0 and temperature range of 4 to 40 °C, with an optimum growth temperature of 28 °C. The resistance of Ni(II) varied from 0.5 to 5 mM. Ni(II) reduction was still observed when nitrate was present in addition to oxygen as a potential electron acceptor. The Ni(II) reduction efficiency was related with the molar ratio of the electron donor to Ni(II). Unlike other dissimilatory metal-reducing bacteria, which oxidizes organic matter with Fe(III) or Mn(IV) as the sole electron acceptor coupled to energy production under facultative anaerobic conditions, this strain used oxygen as an electron acceptor combined with metal reduction. The aerobic metal reduction may relate to a co-metabolic reduction. Transmission electron microscopy images demonstrated that the cells had the ability to accumulate heavy metals, and that the detoxicity mechanism was intracellular metal reduction. These results suggested that the use of Pseudomonas sp. MBR could be promising for toxic heavy metal bioremediation and biological metallurgy.     

Contamination by uranium mine drainages affects fungal growth and interactions between fungal species and strains

The presence of aquatic hyphomycetes has been reported for several heavy metal-contaminated waters. Tolerance probably is one adaptation to coping with heavy metals. To help clarify this issue strains of two species of aquatic hyphomycetes (Tricladium splendens Ingold and Varicosporium elodeae Kegel) were isolated from a reference stream and a stream contaminated with heavy metals and grown on malt extract agar prepared with reference and contaminated water to characterize colony morphology, growth rate, growth inhibition and interaction among species and strains. In V. elodeae the morphology of colonies differed between strains. Colony diameter increased linearly over time with growth rates being lower for strains isolated from contaminated than from reference streams (mostly for V. elodeae). Strains from the contaminated stream grew faster in medium prepared with contaminated water than in medium prepared with reference water, while for strains from the reference stream there was no significant difference in growth rates on the two media. In interacting isolates radial growth toward the opposing colony was generally lower than toward the dish edge. Percentage growth inhibition was higher for isolates in intraspecific interactions (13-37%) than in interspecific interactions (3-27%). However differences in growth inhibition experienced by interacting isolates were observed only in three cases out of 16. The difference between the percentage inhibition caused and experienced by a given isolate was highest in interactions involving isolates with distinct growth rates. Our results suggest that strains from the reference stream tolerate heavy metals while strains from the contaminated stream seem to be adapted to contaminated waters. We hypothesize that in natural environments fungal species-specific limits of tolerance to metal contamination might determine an abrupt or gradual response of the original fungal community to mine pollution giving origin to a poorer fungal community dominated by adapted strains with distinct functional efficiency.     

Quantification of Heavy Metals in Mining Affected Soil and Their Bioaccumulation in Native Plant Species

Several anthropogenic and natural sources are considered as the primary sources of toxic metals in the environment. The current study investigates the level of heavy metals contamination in the flora associated with serpentine soil along the Mafic and Ultramafic rocks northern-Pakistan. Soil and wild native plant species were collected from chromites mining affected areas and analyzed for heavy metals (Cr, Ni, Fe, Mn, Co, Cu and Zn) using atomic absorption spectrometer (AAS-PEA-700). The heavy metal concentrations were significantly (p < 0.01) higher in mine affected soil as compared to reference soil, however Cr and Ni exceeded maximum allowable limit (250 and 60 mg kg^?1, respectively) set by SEPA for soil. Inter-metal correlations between soil, roots and shoots showed that the sources of contamination of heavy metals were mainly associated with chromites mining. All the plant species accumulated significantly higher concentrations of heavy metals as compared to reference plant. The open dumping of mine wastes can create serious problems (food crops and drinking water contamination with heavy metals) for local community of the study area. The native wild plant species (Nepeta cataria, Impatiens bicolor royle, Tegetis minuta) growing on mining affected sites may be used for soil reclamation contaminated with heavy metals.     

Isolation and characterization of heavy-metal-mobilizing bacteria from contaminated soils and their potential in promoting Pb, Cu, and Cd accumulation by Coprinus comatus

The enhanced effect of heavy-metal-mobilizing bacteria on the uptake of Pb, Cu, and Cd by Coprinus comatus from Pb-, Cu-, and Cd-multicontaminated soil was assessed in this study. Thirteen strains, tolerating 800?mg·L(-1) Pb, 200?mg·L(-1) Cu, and 200?mg·L(-1) Cd simultaneously were selected for heavy-metal-solubilizing experiments in soil. The mobilization of heavy metals depended on the characteristics of bacteria and heavy metals. Correlation analysis demonstrated that for Pb solubilization, the acid-producing ability was the most significant factor, while for Cu and Cd, siderophores played a leading role in this process. Four strains, based on their excellent ability to solubilize heavy metal in soil, were applied in pot experiments. The results showed that all strains can promote the growth of C. comatus and meanwhile help mushroom accumulate more heavy metals (Pb, Cd, and Cu). The maximum uptake for total Pb and Cu by C. comatus was observed in inoculations with Bacillus sp. strain JSG1 (2.02- and 2.13-fold, respectively, compared with uninoculated soil), while for Cd, it was recorded in Bacillus sp. strain PB2 treated soil (2.03-fold). Therefore, this work suggests that the mushroom-bacteria interaction can be developed into a novel bioremediation strategy.     

Screening of plant growth promoting traits in heavy metals resistant bacteria: Prospects in phytoremediation

Phytoremediation is considered as a novel environmental friendly technology, which uses plants to remove or immobilize heavy metals. The use of metal-resistant plant growth-promoting bacteria (PGPB) constitutes an important technology for enhancing biomass production as well as tolerance of the plants to heavy metals. In this study, we isolated twenty seven (NF1-NF27) chromium resistant bacteria. The bacteria were tested for heavy metals (Cr, Zn, Cu, Ni, Pb and Co) resistance, Cr(VI) reduction and PGPB characters (phosphate solubilization, production of IAA and siderophores). The results showed that the bacterial isolates resist to heavy metals and reduce Cr(VI), with varying capabilities. 37.14% of the isolates have the capacity of solubilizing phosphate, 28.57% are able to produce siderophores and all isolates have the ability to produce IAA. Isolate NF2 that showed high heavy metal resistance and plant growth promotion characteristics was identified by 16S rDNA sequence analysis as a strain of Cellulosimicrobium sp.. Pot culture experiments conducted under greenhouse conditions showed that this strain was able to promote plant growth of alfalfa in control and in heavy metals (Cr, Zn and Cu) spiked soils and increased metal uptake by the plants. Thus, the potential of Cellulosimicrobium sp. for both bioremediation and plant growth promotion has significance in the management of environmental pollution.     

Genome Sequence Analysis of the Naphthenic Acid Degrading and Metal Resistant Bacterium Cupriavidus gilardii CR3

Cupriavidus sp. are generally heavy metal tolerant bacteria with the ability to degrade a variety of aromatic hydrocarbon compounds, although the degradation pathways and substrate versatilities remain largely unknown. Here we studied the bacterium Cupriavidus gilardii strain CR3, which was isolated from a natural asphalt deposit, and which was shown to utilize naphthenic acids as a sole carbon source. Genome sequencing of C. gilardii CR3 was carried out to elucidate possible mechanisms for the naphthenic acid biodegradation. The genome of C. gilardii CR3 was composed of two circular chromosomes chr1 and chr2 of respectively 3,539,530 bp and 2,039,213 bp in size. The genome for strain CR3 encoded 4,502 putative protein-coding genes, 59 tRNA genes, and many other non-coding genes. Many genes were associated with xenobiotic biodegradation and metal resistance functions. Pathway prediction for degradation of cyclohexanecarboxylic acid, a representative naphthenic acid, suggested that naphthenic acid undergoes initial ring-cleavage, after which the ring fission products can be degraded via several plausible degradation pathways including a mechanism similar to that used for fatty acid oxidation. The final metabolic products of these pathways are unstable or volatile compounds that were not toxic to CR3. Strain CR3 was also shown to have tolerance to at least 10 heavy metals, which was mainly achieved by self-detoxification through ion efflux, metal-complexation and metal-reduction, and a powerful DNA self-repair mechanism. Our genomic analysis suggests that CR3 is well adapted to survive the harsh environment in natural asphalts containing naphthenic acids and high concentrations of heavy metals.     

Chromate removal by yeasts isolated from sediments of a tanning factory and a mine site in Argentina

Twenty-one yeast-like microorganisms were isolated from tannery effluents and from a nickel–copper mine in Argentina. They were tested for their Cu(II), Ni(II), Cd(II) and Cr(VI) tolerance in qualitative assays on solid medium. Three isolates were selected for their multiple tolerance to the different heavy metals and highest tolerance to Cr(VI). According to morphological and physiological analysis and 26S rDNA D1/D2 domain sequences the isolates were characterized as: Lecythophora sp. NGV-1, Candida sp. NGV-9 and Aureobasidium pullulans VR-8. Resistance of the three strains to high Cr(VI) concentrations and their ability to remove Cr(VI) were assessed using YNB-glucose medium supplemented with 0.5 and 1 mM Cr(VI). Chromate removal activity was estimated by measuring remaining Cr(VI) concentration in the supernatant using the colorimetric 1,5-diphenylcarbazide method and total chromium was determined by flame atomic absorption spectroscopy. The results indicate that the initial Cr(VI) concentration negatively influenced growth and the specific growth rate but stimulated the metabolic activity of the three strains; resistance to Cr(VI) by these strains was mainly due to reduction of Cr(VI) rather than chromium bioaccumulation. This study showed the potential ability of these strains as tools for bioremediation of Cr(VI) from contaminated sites.     

Modeling of phytoextraction efficiency of microbially stimulated Salix dasyclados L. in the soils with different speciation of heavy metals

Bioaugmentation of soils with selected microorganisms during phytoextraction can be the key solution for successful bioremediation and should be accurately calculated for different physicochemical soil properties and heavy metal availability to guarantee the universality of this method. Equally important is the development of an accurate prediction tool to manage phytoremediation process. The main objective of this study was to evaluate the role of three metallotolerant siderophore-producing Streptomyces sp. B1–B3 strains in the phytoremediation of heavy metals with the use of S. dasyclados L. growing in four metalliferrous soils as well as modeling the efficiency of this process based on physicochemical and microbiological properties of the soils using artificial neural network (ANN) analysis. The bacterial inoculation of plants significantly stimulated plant biomass and reduced oxidative stress. Moreover, the bacteria affected the speciation of heavy metals and finally their mobility, thereby enhancing the uptake and bioaccumulation of Zn, Cd, and Pb in the biomass. The best capacity for phytoextraction was noted for strain B1, which had the highest siderophore secretion ability. Finally, ANN model permitted to predict efficiency of phytoextraction based on both the physicochemical properties of the soils and the activity of the soil microbiota with high precision.     

Isolation and Characterization of A Metal-resistant Pseudomonas Aeruginosa Strain

Summary The use of microorganisms to remove heavy metals from industrial effluent is an area of extensive research and development. Attempts have been made to isolate and characterize metal-resistant microorganisms from treated oil mill industry effluent wastewater samples. The metal-resistant organisms that showed values of minimum inhibitory concentration towards metals (Cd, Cr, Ni and Pb) ranging from 100 to 800 ppm level were screened. A potent metal-resistant organism, isolate BC15 from the wastewater samples was tentatively identified as Pseudomonas sp. Detailed analysis of morphological, biochemical and 16S rDNA sequence of the isolate revealed that it is closely related to Pseudomonas aeruginosa (94%). Pseudomonas BC15 was capable of absorbing 93% Ni, 65% Pb, 50% Cd and 30% Cr within 48 h from the medium containing 100 mg of each heavy metal per liter. The multiple metal tolerance of this strain was also associated with resistance to antibiotics such as ampicillin, tetracycline, chloramphenicol, erythromycin, kanamycin and streptomycin.     

Biofilm establishment and heavy metal removal capacity of an indigenous mining algal-microbial consortium in a photo-rotating biological contactor

An indigenous mining algal-microbial consortium was immobilised within a laboratory-scale photo-rotating biological contactor (PRBC) that was used to investigate the potential for heavy metal removal from acid mine drainage (AMD). The microbial consortium, dominated by Ulothrix sp., was collected from the AMD at the Sar Cheshmeh copper mine in Iran. This paper discusses the parameters required to establish an algal-microbial biofilm used for heavy metal removal, including nutrient requirements and rotational speed. The PRBC was tested using synthesised AMD with the multi-ion and acidic composition of wastewater (containing 18 elements, and with a pH of 3.5?±?0.5), from which the microbial consortium was collected. The biofilm was successfully developed on the PRBC's disc consortium over 60?days of batch-mode operation. The PRBC was then run continuously with a 24?h hydraulic residence time (HRT) over a ten-week period. Water analysis, performed on a weekly basis, demonstrated the ability of the algal-microbial biofilm to remove 20-50?% of the various metals in the order Cu?>?Ni?>?Mn?>?Zn?>?Sb?>?Se?>?Co?>?Al. These results clearly indicate the significant potential for indigenous AMD microorganisms to be exploited within a PRBC for AMD treatment.     

Culturable Heavy Metal-Resistant and Plant Growth Promoting Bacteria in V-Ti Magnetite Mine Tailing Soil from Panzhihua,China

To provide a basis for using indigenous bacteria for bioremediation of heavy metal contaminated soil, the heavy metal resistance and plant growth-promoting activity of 136 isolates from V-Ti magnetite mine tailing soil were systematically analyzed. Among the 13 identified bacterial genera, the most abundant genus was Bacillus (79 isolates) out of which 32 represented B. subtilis and 14 B. pumilus, followed by Rhizobium sp. (29 isolates) and Ochrobactrum intermedium (13 isolates). Altogether 93 isolates tolerated the highest concentration (1000 mg kg⁻¹) of at least one of the six tested heavy metals. Five strains were tolerant against all the tested heavy metals, 71 strains tolerated 1,000 mg kg⁻¹ cadmium whereas only one strain tolerated 1,000 mg kg⁻¹ cobalt. Altogether 67% of the bacteria produced indoleacetic acid (IAA), a plant growth-promoting phytohormone. The concentration of IAA produced by 53 isolates was higher than 20 µg ml⁻¹. In total 21% of the bacteria produced siderophore (5.50–167.67 µg ml⁻¹) with two Bacillus sp. producing more than 100 µg ml⁻¹. Eighteen isolates produced both IAA and siderophore. The results suggested that the indigenous bacteria in the soil have beneficial characteristics for remediating the contaminated mine tailing soil.     

Removal of heavy metals by an enriched consortium

An enriched consortium obtained from lake-sediment was developed for the removal of heavy metals such as Cu, Pb, Cr, Ni, and Zn from heavy metal-contaminated water. The removal efficiency of heavy metals in a shaking condition was generally higher than that in the static state. After the fifteenth enrichment with assorted heavy metals, the removal efficiencies in the shaking and static condition at an average concentration of 100 mg/L of each heavy metal were approximately 99 approximately 100% and 95 approximately 100%, respectively, depending on the type of heavy metal. An aerobically grown, pure culture isolated from an enriched culture was analyzed by 16S rRNA sequencing and identified as Ralstonia sp. HM-1. This strain was found to remove various heavy metals with an efficiency of approximately 97 approximately 100% at an average concentration of 200 mg/L of each heavy metal.