Bacterial leaching of manganese ores

Leaching of various types of ores, containing 12-30% manganese, by the thiobacterium Acidithiobacillus ferrooxidans was studied. Leaching of reduced ores (manganocalcite and manganiferous limestone) was mediated mainly by degradation of manganiferous minerals (by sulfuric acid produced in the course of bacterial oxidation of pyrite or sulfur). Bacterial treatment of the ores for 144 and 192 h allowed solubilization of 96-98% of manganese. Inoculation of bacteria into pulp with pyrite increased the rate of leaching of oxide ore (psilomelane) by 37%, and the degree of its extraction within 180 h increased from 80 to 97%.     

Importance of Extracellular Polymeric Substances from Thiobacillus ferrooxidans for Bioleaching

Leaching bacteria such as Thiobacillus ferrooxidans attach to pyrite or sulfur by means of extracellular polymeric substances (EPS) (lipopolysaccharides). The primary attachment to pyrite at pH 2 is mediated by exopolymer-complexed iron(III) ions in an electrochemical interaction with the negatively charged pyrite surface. EPS from sulfur cells possess increased hydrophobic properties and do not attach to pyrite, indicating adaptability to the substrate or substratum.     

Physiology and kinetics of manganese‐reducing bacillus polymyxa strain D1 isolated from manganiferous silver ore

Manganese‐reducing bacteria were isolated from a manganiferous silver ore mining site using enrichment procedures. The most rapid Mn(IV) reducer was identified as Bacillus polymyxa and was designated as strain D1. Isolate D1 has no growth‐factor requirements and is mesophilic and neutrophilic. D1 respires glucose aerobically, under which conditions cyanide is bactericidal. Nonfermentable substrates such as lactate, acetate, citrate, and succinate cannot serve as sole carbon sources. D1 ferments glucose anaerobically, producing acetic acid, ethanol, and butanediol as major metabolic end products. Both anaerobic conditions and direct physical contact with pyrolusite (MnO₂) particles were necessary for manganese reduction. Strain D1 is unique in that manganese serves as an ancillary electron acceptor during anaerobic fermentation. Kinetic experiments showed that D1 reduced manganese three to five times as rapidly as the widely studied Mn(IV)/Fe(III)‐reducing microorganisms Shewanella putrefaciens MR‐1 and Shewanella putrefa‐ciens sp. 200. Strain D1 is capable of liberating silver via the reductive dissolution of refractory manganiferous ores.     

功能菌群耦合黄铁矿浸出软锰矿的研究

【目的】将3种不同来源的环境样品混合后接种至含1%黄铁矿和1%软锰矿的培养基中进行富集培养,初步得到有一定浸矿功能的混合微生物菌群。【方法】菌群继续用于黄铁矿和低品位软锰矿共同浸出,设置未接种的体系作为对照。【结果】对浸出过程中菌群结构的变化、pH、锰浸出率和浸出残渣的成分进行分析,结果发现接种过微生物菌群的浸出体系在反应15 d后,锰浸出率达到92.48%,远高于未接菌对照组的40.34%;菌群中Thiomonas sp.所占比例从最初的2%上升到浸出结束时的93%。实验组的pH从最初的4.0下降到2.5;X射线衍射(XRD)分析发现,通过生物作用浸出的残渣中含有黄钾铁矾,说明生物代谢产生了大量的硫酸。【结论】证明微生物在两矿浸出过程中通过促进黄铁矿解离,维持体系低pH等作用加速反应的进行。结果为进一步研究微生物浸矿的作用机制和开发低品位锰矿的生物浸出工艺打下了基础。     

锰氧化细菌的分离鉴定及其锰氧化特性的分析

利用选择性培养基对锰矿样品进行分离、筛选, 得到一株高效锰氧化细菌(MN1405)。经形态特征、生理生化特征以及16S rRNA 基因序列分析, 将菌株MN1405 鉴定为Arthrobacter echigonensis。在培养条件下, MN1405 对培养基中的锰离子去除率可达93.38%, 且其培养所获得的培养液也具有良好的除锰效果。     

Manipulation of pyrite colonization and leaching by iron-oxidizing <Emphasis Type="Italic">Acidithiobacillus</Emphasis> species

In this study, the process of pyrite colonization and leaching by three iron-oxidizing Acidithiobacillus species was investigated by fluorescence microscopy, bacterial attachment, and leaching assays. Within the first 4–5 days, only the biofilm subpopulation was responsible for pyrite dissolution. Pyrite-grown cells, in contrast to iron-grown cells, were able to oxidize iron(II) ions or pyrite after 24 h iron starvation and incubation with 1 mM H₂O₂, indicating that these cells were adapted to the presence of enhanced levels of reactive oxygen species (ROS), which are generated on metal sulfide surfaces. Acidithiobacillus ferrivorans SS3 and Acidithiobacillus ferrooxidans R1 showed enhanced pyrite colonization and biofilm formation compared to A. ferrooxidans ^T. A broad range of factors influencing the biofilm formation on pyrite were also identified, some of them were strain-specific. Cultivation at non-optimum growth temperatures or increased ionic strength led to a decreased colonization of pyrite. The presence of iron(III) ions increased pyrite colonization, especially when pyrite-grown cells were used, while the addition of 20 mM copper(II) ions resulted in reduced biofilm formation on pyrite. This observation correlated with a different extracellular polymeric substance (EPS) composition of copper-exposed cells. Interestingly, the addition of 1 mM sodium glucuronate in combination with iron(III) ions led to a 5-fold and 7-fold increased cell attachment after 1 and 8 days of incubation, respectively, in A. ferrooxidans ^T. In addition, sodium glucuronate addition enhanced pyrite dissolution by 25 %.     

Dependence of the Phenotypic Characteristics of <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis> on the Physical,Chemical, and Electrophysical Properties of Pyrites

Comparison of Acidithiobacillus ferrooxidans strains TFV-1 and TFBk with respect to their capacity to oxidize pyrite 1, with an electron-type (n-type) conductivity, or pyrite 2, with hole-type (p-type) conductivity, showed that, at a pulp density of 1%, both before and after its adaptation to the pyrites, strain TFBk, isolated from a substrate with a more complex mineral composition, grew faster and oxidized the pyrites of both conductivity types more efficiently than strain TFV-1, which was isolated from a mineralogically simple ore. At a pulp density of 3–5%, the oxidation of pyrite 2 by strain TFV-1 and both of the pyrites by strain TFBk began only after an artificial increase in Eh to 600 mV. If the pulp density was increased gradually, strain TFBk could oxidize the pyrites at its higher values than strain TFV-1, with the rate of pyrite 2 oxidation being higher than that of pyrite 1. During chemical oxidation of both of the pyrites, an increase was observed in the absolute values of the coefficients of thermoelectromotive force (K_TEMF); during bacterial-chemical oxidation, the K_TEMF of pyrite 1 changed insignificantly, whereas the K_TEMF of pyrite 2 decreased.     

Effect of anions on selective solubilization of zinc and copper in bacterial leaching of sulfide ores

Bacterial leaching of sulfide ores using Thiobacillus ferrooxidans, Thiobacillus thiooxidans, or a combination of the two was studied at various concentrations of specific anions. Selective zinc and copper solubilization was obtained by inhibiting iron oxidation without affecting sulfur/sulfide oxidation. Phosphate reduced iron solubilization from a pyrite (FeS(2))-sphalerite (ZnS) mixture without significantly affecting zinc solubilization. Copper leaching from a chalcopyrite (CuFeS(2))-sphalerite mixture was stimulated by phosphate, whereas chloride accelerated zinc extraction. In a complex sulfide ore containing pyrite, chalcopyrite, and sphalerite, both phosphate and chloride reduced iron solubilization and increased copper extraction, whereas only chloride stimulated zinc extraction. Maximum leaching obtained was 100% zinc and 50% copper. Time-course studies of copper and zinc solubilization suggest the possibility of selective metal recovery following treatment with specific anions.     

Rate of Pyrite Bioleaching by Thiobacillus ferrooxidans: Results of an Interlaboratory Comparison

Ten laboratories participated in an interlaboratory comparison of determination of bioleaching rates of a pyrite reference material. A standardized procedure and a single strain of Thiobacillus ferrooxidans were used in this study. The mean rate of bioleaching of the pyrite reference material was 12.4 mg of Fe per liter per h, with a coefficient of variation (percent relative standard deviation) of 32% as determined by eight laboratories. These results show the precision among laboratories of the determination of rates of pyrite bioleaching when a standard test procedure and reference material are used.     

Selective Inhibition of the Oxidation of Ferrous Iron or Sulfur in Thiobacillus ferrooxidans

The oxidation of either ferrous iron or sulfur by Thiobacillus ferrooxidans was selectively inhibited or controlled by various anions, inhibitors, and osmotic pressure. Iron oxidation was more sensitive than sulfur oxidation to inhibition by chloride, phosphate, and nitrate at low concentrations (below 0.1 M) and also to inhibition by azide and cyanide. Sulfur oxidation was more sensitive than iron oxidation to the inhibitory effect of high osmotic pressure. These differences were evident not only between iron oxidation by iron-grown cells and sulfur oxidation by sulfur-grown cells but also between the iron and sulfur oxidation activities of the same iron-grown cells. Growth experiments with ferrous iron or sulfur as an oxidizable substrate confirmed the higher sensitivity of iron oxidation to inhibition by phosphate, chloride, azide, and cyanide. Sulfur oxidation was actually stimulated by 50 mM phosphate or chloride. Leaching of Fe and Zn from pyrite (FeS₂) and sphalerite (ZnS) by T. ferrooxidans was differentially affected by phosphate and chloride, which inhibited the solubilization of Fe without significantly affecting the solubilization of Zn.     

Dependence of the Genotypic Characteristics of <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis> on the Physical,Chemical, and Electrophysical Properties of Pyrites

This study focused on the effect of physical, chemical, and electrophysical properties of two pyrites, pyrite 1, which had electron-type (n-type) conductivity, and pyrite 2, with hole-type (p-type) conductivity, on the genotypic characteristics of Acidithiobacillus ferrooxidans strains TFV-1 and TFBk, which were isolated from different substrates. After the adaptation of the strains to the pyrites at a pulp density of 1%, pulsed-field electrophoresis revealed changes in the chromosomal DNA of strain TFV-1 adapted to pyrite 1, and strain TFBk adapted to either of the pyrite types. In pyrite-adapted strain TFBk, the plasmid composition was the same as after growth on a medium containing ferrous iron, whereas, in strain TFV-1, changes in plasmid sizes or both in plasmid sizes and plasmid number occurred. After an increase in the density of the pyrite 2 pulp from 1 to 10%, the plasmid number increased from three to four, and, after an increase in the density of the pyrite 1 pulp from 1 to 7%, the plasmid number increased from two to six.     

Bacterial Oxidation of Refractory Sulfide Ores for Gold Recovery

Abstract

The microbiological leaching of refractory sulfide ores (pyrite, arsenopyrite) for recovery of gold is reviewed in this article. The underlying physiological, biochemical, and genetic fundamentals of the bacteria involved (Thiobacillus and Sulfolobus spp.) are complex and have yet to be elucidated in depth. The chemistry of acid and biological leaching of pyrite and arsenopyrite minerals is also complex, and many of the individual reactions are not known in detail. Bacterial leaching is discussed in relation to chemical speciation at acid pH values. Attempts to develop models for a better understanding of bioleaching processes are summarized. The importance of pH, redox potential, temperature, sulfur balance, and toxic metals is evaluated for optimizing conditions for bacterial activity. Gold is finely disseminated in refractory sulfide ores, thereby decreasing Au recoveries upon conventional cyanidation for gold dissolution. In the bioleaching process, bacteria remove the sulfide minerals by oxidative dissolution and thus expose Au to extraction with cyanide solution. Stirred tank reactors appear most suited for this biological leaching process. The overall oxidation of the sulfides is an important variable for gold recovery. Pilot- and commercial-scale bioleaching processes for gold-containing pyrite and arsenopyrite ores are reviewed. This application of mineral biotechnology competes favorably with pressure leaching and roasting processes, both of which are problematic and energy-intensive alternatives for pretreatment of auriferous pyrite/arsenopyrite ores.     

Soil retention of 15N in a simulated N deposition study: effects of live plant and soil organic matter content

Background and aims

The impacts of atmospheric nitrogen (N) deposition on terrestrial ecosystem processes remain controversial, mostly because of the uncertainty regarding the fates of deposited N. We conducted a 16-week simulated deposition study to experimentally trace N in a greenhouse plant-soil system.

Methods

Using a two-way factorial design, we added (¹⁵NH₄)₂SO₄ solution twice a week to pots containing different soil organic matter (SOM) content and with or without a live plant (Salix dasyclados). The recoveries of ¹⁵N in soil, plant biomass, and leaching solution were quantified.

Results

We found most ¹⁵N was retained in soil (18.0–59.2%), with significantly more ¹⁵N recovered from high-SOM soils than from low-SOM soils. Plant presence significantly increased ¹⁵N retention in soil. Plant biomass accounted for 10–20% of the ¹⁵N input, with proportionally more ¹⁵N assimilated when plants were grown in low-SOM soils. Leaching loss of ¹⁵N was relatively low (10–17%).

Conclusion

Our study suggests that SOM content and plant presence significantly affect the fates of deposited N. Indeed, N would be preferentially retained in soils with high SOM content and live plant, while plants would assimilate more deposited N when grown in low SOM soils. Global biogeochemical models thus need to incorporate such soil-specific N retention and plant N assimilation.     

Loci underlying resistance to manganese toxicity mapped in a soybean recombinant inbred line population of ‘Essex2019; x ‘Forrest’

Resistance to manganese toxicity is associated with some soybean (Glycine max) cultivars grown on acidic soils or in hydroponics. Previously random amplified polymorphic DNA (RAPD) markers had seemed to identify 4 quantitative trait loci (QTL), regions that might underlie resistance to manganese toxicity in a recombinant inbred line (RIL) population derived from ‘Essex’ x ‘Forrest’. Our objective was to identify microsatellite markers linked to these, or additional, QTL for resistance to manganese toxicity in a separate assay. Two hundred and forty microsatellite markers and 100 RILs were used to construct a map. The response of five plants per genotype to manganese was measured by leaf chlorosis (scored from 0–5) and root necrosis (scored from 0–5) from 7–28 days after treatment with 125 μM of manganese in hydroponics. The experiment was repeated. ANOVA and MapMaker/QTL were used to identify regions underlying the responses. Three genomic regions on different linkage groups were found to contain QTL for resistance to necrosis during manganese toxicity. The regions located on linkage groups C2 (BARC__S att291),I(BARC__S att239)andG(OP__O EO2)wereeachsignificantlyassociated(P<0.005, R ²=20%) with root necrosis at 7 days after treatment. The regions all derived the beneficial allele from Essex. One of the previously identified RAPD associated root necrosis QTL was identified in this new study. However, no QTL for leaf chlorosis were detected (P<0.005) and none of the RAPD identified leaf chlorosis QTL could be identified. We conclude that root and leaf resistance to manganese toxicity are environmentally sensitive quantitative traits determined by separate loci of different number and magnitude of effect.     

Reactive oxygen species generated in the presence of fine pyrite particles and its implication in thermophilic mineral bioleaching

In the tank bioleaching process, maximising solid loading and mineral availability, the latter through decreasing particle size, are key to maximising metal extraction. In this study, the effect of particle size distribution on bioleaching performance and microbial growth was studied through applying knowledge based on medical geology research to understand the adverse effects of suspended fine pyrite particles. Small-scale leaching studies, using pyrite concentrate fractions (106–75, 75–25, ?25 μm fines), were used to confirm decreasing performance with decreasing particle size (D ₅₀ <40 μm). Under equivalent experimental conditions, the generation of the reactive oxygen species (ROS), hydrogen peroxide and hydroxyl radicals from pyrite was illustrated. ROS generation measured from the different pyrite fractions was found to increase with increasing pyrite surface area loading (1.79–74.01 m² L^?1) and Fe²⁺ concentration (0.1–2.8 g?L^?1) in solution. The highest concentration of ROS was measured from the finest fraction of pyrite (0.85 mM) and from the largest concentration of Fe²⁺ (0.78 mM). No ROS was detected from solutions containing only Fe³⁺ under the same conditions tested. The potential of ROS to inhibit microbial performance under bioleaching conditions was demonstrated. Pyrite-free Sulfolobus metallicus cultures challenged with hydrogen peroxide (0.5–2.5 mM) showed significant decrease in both cell growth and Fe²⁺ oxidation rates within the concentration range 1.5–2.5 mM. In combination, the results from this study suggest that conditions of large pyrite surface area loading, coupled with high concentrations of dissolved Fe²⁺, can lead to the generation of ROS, resulting in oxidative stress of the microorganisms.     

Manganese‐Rich layers in calcareous deposits along the Western shore of the Dead Sea May have a bacterial origin

Spore formers were detected in samples of calcareous crust with manganiferous laminations and in water from the Dead Sea in Israel. They were able to grow in media made with fresh water but not with synthetic Dead Sea water. Some of these spore formers were also able to oxidize Mn (II) in fresh water. No other bacteria capable of both growth and Mn(II) oxidation in hypersaline media prepared with synthetic Dead Sea water were found in the samples. However, bacteria capable of growth and Mn(II) oxidation in fresh water media were detected in water and sediment from fresh water springs at Ein Feshkha and in lake water and sediment from the beach near Wadi Kidron. Both sites are located on the western shore of the lake. These findings suggest that the manganiferous laminations in calcareous crusts and concretions in the Dead Sea along its western shore may have originated, at least in part, from manganese oxide formed by bacteria in fresh water environments on shore and washed into the lake in runoff, with subsequent incorporation into the crusts and concretions.     

九州虫草菌丝体对Mn的耐性及富集

重金属耐性真菌的研究是生物修复的重要研究内容。本文研究了九州虫草（Cordyceps kyusyuensis）对于Mn的耐性及富集。在液体培养基中添加不同浓度（0—60 g/L）的Mn离子,测定其菌丝生物量、菌丝Mn含量、菌丝抗氧化酶活性和过氧化水平以及菌体细胞离子交换量、Mn在细胞中的分布的变化情况。实验结果表明九州虫草菌丝生物量与Mn浓度呈显著负相关,Mn浓度60 g/L为九州虫草菌丝生长极限浓度。菌丝中Mn含量随培养基中Mn浓度的增大而显著升高,10 g/L Mn时,菌丝细胞中Mn积累量达到细胞干重的1.0013%。九州虫草菌丝中过氧化产物丙二醛（MDA）、可溶性蛋白（SP）含量、可溶性糖浓度与培养基中Mn浓度呈负相关,实验组与对照组差异显著。抗氧化酶（过氧化氢酶（CAT）、过氧化物酶（POD）、超氧化物歧化酶（SOD））活性随着培养基中Mn浓度增大而显著升高,但变化趋势不同。九州虫草菌丝细胞不可溶性组分中Mn的量（91.51%—98.6%）显著高于可溶部分（1.40%—8.49%）。九州虫草菌丝细胞壁离子交换量（CEC）随着培养基中Mn浓度的升高变化不明显。说明在九州虫草菌丝对Mn的富集过程中,其细胞壁、细胞膜和细胞器对于Mn结合发挥了主要作用,细胞质中可溶性成分对Mn的结合发挥次要作用。在Mn的胁迫下,增强抗氧化酶系统的协同作用以清除大量自由基是细胞对锰耐性的重要机制。     

Manganese-induced apoptosis in rat myocytes

Manganese can be toxic to the heart, causing dysfunction following long exposure. In our experiments, we examined the cytotoxicity of manganese in neonatal rat ventricular myocytes (NRVM) by MTT assays in vitro. Results showed that after incubation in the different concentrations of manganese for 24 h, apparent cytotoxicity was observed. At 500, 1000, and 1500 2 microM of manganese, the percentage of cell viability dropped to 82% +/- 6.13, 78% +/- 5.28, and 66% +/- 4.22, respectively. When cells were treated for 48 h, all concentrations tested exerted toxic effect; especially from 500 to 1500 microM the cell viability dropped from 67% +/- 4.84 to 37% +/- 3.25. Apoptosis in NRVM was then examined by flow cytometry. Results showed that the percentage of apoptotic cells treated with 500 microM of manganese for 24 h increased from 4% +/- 0.84 to 7% +/- 1.16. After 48 h of incubation, this percentage increased to 11% +/- 0.91. There was no significant difference between control groups (0 microM manganese) after 24 and 48 h incubation. The morphological changes of NRVM nuclei were visualized with the fluorescent DNA-binding dye Hoechst33342 after incubation in 500 microM of manganese for 48 h. Compared with normal nuclei, apoptotic nuclei showed the typical features of fragmentation and condensation. To investigate whether there are any apoptotic gene expression changes during apoptosis, we examined the expression level of Bcl-2, Bax, and P53 mRNAs after treatment with 500 microM of manganese for 48 h. The Bcl-2 mRNA expression decreased while the expression of Bax as well as P53 mRNAs increased. These results suggested that manganese cytotoxicity on NRVM could induce apoptosis in NRVM cells. The apoptosis process might involve, and be promoted by, the changes of the expression levels of P53, Bcl-2, and Bax proteins.