高效浸矿混合菌种的筛选及其在浸矿过程中的微生物群落结构演变

为了筛选可高效浸出低品位硫化矿的混合菌株,从10个典型的硫化矿矿区的酸性矿坑水中分离富集到混合菌株。以浸出率作为筛选的主要标准,筛选后的高效菌株,利用群落基因组芯片分析混合菌群的组成和过程变化。浸矿持续进行24d后,采集自江西德兴银山铅锌矿的混合菌浸出率最高,为68.89%。群落基因组芯片结果表明银山菌群包含9种菌,可被分为6类,在浸出过程中群落始终在变化。该研究为加速浸出和了解浸出过程中群落演替提供了一个较好的方法。     

Comparative study on bioleaching of two different types of low-grade copper tailings by mixed moderate thermophiles

The bioleaching of two different types of low-grade copper tailings, acid-leaching tailings (ALT) and copper flotation tailings (CFT) by mixed moderate thermophiles, and the variation of mineralogical and microbiological characteristics during their dissolution processes were comparatively investigated. Results showed that bioleaching behaviors of the two types of tailings were significantly different. In ALT bioleaching, lower redox potential, higher [Fe³⁺]/[Fe²⁺] ratio and higher cell density in solution were obtained. These resulted in higher total copper, primary copper sulfide and secondary copper sulfide extractions, compared with CFT bioleaching. X-ray diffraction analysis suggested that gypsum and some metal organic complexes were detected in CFT bioleaching, which could cause the sluggish oxidation of sulphide minerals. The shifts of microbial community in the leachates and leaching residues varied greatly between ALT and CFT bioleaching. The percentage of iron-oxidizing bacteria in ALT bioleaching was higher than that of CFT, but the sulfur-oxidizing bacteria percentage was the opposite. The archaeon F. thermophilum L1 was detected in ALT but not in CFT.     

Dominance of Acidithiobacillus at ore surface of Zijinshan commercial low-grade copper bioleaching heap

The microbial community structure in the ore surface of Zijinshan commercial low-grade copper bioleaching heap was investigated by 16S rRNA gene clone library.For both bacteria and Archaea,105 clones were sequenced.The dominant bacteria species present in the ore surface were Acidithiobacillus and Leptospirillum,accounting for 51.42% and 48.57%,respectively. However,for the Archaea,only one operational taxonomic unit(OUT)belonged to Ferroplasma acidiphilum.These results indicate that function of genus Ac...     

微生物浸出Pb-Zn-Sn黄铜矿及其生物群落的演替分析

研究了Pb-Zn-Sn黄铜矿精矿在混合中度嗜热微生物槽浸过程中的细菌群落结构变化,并监测浸出体系中金属离子浓度、溶液电位、溶液pH值变化,通过聚合酶链式反应-限制性片段长度多态性（PCR-RFLP）技术分析微生物群落的结构变化。结果表明,最终铜浸出率高达85.6%,在浸出前期,Acidithiobacillus caldus为优势群落,从第18天开始到浸出结束,Sulfobacillus thermosulfidooxidans为优势群落,但Leptospirillum ferriphilum丰度变化较小。试验结果表明,适当较高的溶液电位和合适的铁离子浓度对黄铜矿精矿的生物浸出作用很关键。     

Dominance of Acidithiobacillus at ore surface of Zijinshan commercial low-grade copper bioleaching heap

The microbial community structure in the ore surface of Zijinshan commercial low-grade copper bioleaching heap was investigated by 16S rRNA gene clone library. For both bacteria and Archaea, 105 clones were sequenced. The dominant bacteria species present in the ore surface were Acidithiobacillus and Leptospirillum, accounting for 51,42% and 48.57%, respectively. However, for the Archaea, only one operational taxonomic unit （OUT） belonged to Ferroplasma acidiphilum. These results indicate that function of genus Acidithiobacillus in the commercial low-grade copper bioleaching heap may be underestimated. More detailed and quantitative information on microbial community structure over time are now under investigation.     

元素硫对黄铜矿生物浸出行为及群落结构的影响(英文)

研究3种典型铁/硫代谢菌—Acidithiobacillus ferrooxidans,Leptospirillum ferriphilum及Acidithiobacillus thiooxidans混合浸出黄铜矿过程中铁/硫氧化活性、群落结构(PCR-RFLP)的变化,以及不同浓度的元素硫对其影响。结果发现,加入3.193g/L元素硫能促进细菌的表观硫氧化活性,改变浸矿体系的群落结构,并进一步影响钝化层的形成、金属离子的溶出,其浸出率(71%)较未添加硫的(67%)有一定程度的提高。而过量的元素硫会抑制铜的浸出(浸出率44%)。     

Research progress in biohydrometallurgy of rare metals and heavy nonferrous metals with an emphasis on China

In the past decade, progress in the field of biohydrometallurgy had been significant. A total of 17 novel biomining microorganisms were discovered, and eight copper heap bioleaching plants and 11 gold biooxidation plants were established or expanded. In this review, it was summarized the physiological properties of the newly isolated biomining microorganisms and three novel microbial ecological methods for studying microbial community dynamics and structure. In addition, biohydrometallurgy research on rare metals such as uranium,molybdenum, tellurium, germanium, indium, and secondary rare metal resources, as well as heavy nonferrous metals such as copper, nickel, cobalt, and gold has been reviewed, with an emphasis on China. In future, further studies on bioleaching of chalcopyrite, rare metals, secondary resources from waste, and environmental pollution caused by resource utilization are necessary.     

细粒铜尾矿生物柱浸体系下细菌群落对潜在的铺矿形式的响应

比较研究分层铺筑(LM)、聚团铺筑(AM)和烧结球团铺筑(PM)三种铺矿形式对铜浮选尾矿生物柱浸的影响.解析尾矿浸出后期三组浸矿体系下游离、吸附、弱吸附和强吸附微生物的群落结构.在AM浸出组,用于尾矿聚团的块状硫化矿的添加使浸出溶液pH值降低,三价铁离子浓度升高和微生物快速吸附.相比于LM(54.6％)和PM(43.9...     

混合高温菌浸出黄铜矿及浸出过程中微生物群落的演替

研究3株极端嗜热古菌(金属硫叶菌,Sulfolobus metallicus JCM 9184;瑟杜生金属球菌,Metallosphaera sedula JCM 9185和万座酸菌,Acidianus manzaensis YN25)在不同起始pH值和不同温度条件下对黄铜矿的混合浸出,并对浸矿过程中混合菌群落的动态演替进行分析.结果表明:在起始pH 1.5时的铜浸出率明显高于在起始pH 2.5时的铜浸出率,而65 ℃条件下的铜浸出率高于75 ℃时的铜浸出率.利用限制性长度多态性(RFLP)分析65 ℃、起始pH 1.5条件下的微生物群落演替,结果显示:在黄铜矿的浸出前期Sulfolobus metallicus是占据优势的菌种,而到后期Acidianus manzaensis的比例则会上升,并最后取代Sulfolobus metallicus成为优势种.     

Biodiversity and interactions of acidophiles:Key to understanding and optimizing microbial processing of ores and concentrates

Mining companies have become increasingly aware of the potential of microbiological approaches for recovering base and precious metals from low-grade ores, and for remediating acidic, metal-rich wastewaters that drain from both operating and abandoned mine sites. Biological systems offer a number of environmental and （sometimes） economical advantages over conventional approaches, such as pyrometallurgy, though their application is not appropriate in every situation. Mineral processing using micro-organisms has been exploited for extracting gold, copper, uranium and cobalt, and current developments are targeting other base metals. Recently, there has been a great increase in our knowledge and understanding of both the diversity of the microbiology of biomining environments, and of how the microorganisms interact with each other. The results from laboratory experiments which have simulated both stirred tank and heap bioreactor systems have shown that microbial consortia are more robust than pure cultures of mineral-oxidizing acidophiles, and also tend to be more effective at bioleaching and bio-oxidizing ores and concentrates. The paper presented a concise review of the nature and interactions of microbial consortia that are involved in the oxidation of sulfide minerals, and how these might be adapted to meet future challenges in biomining operations.     

生物因素对次生硫化铜矿堆浸过程动力学的影响

假定生物浸出过程细菌的作用是间接作用,以实验室柱浸模拟次生硫化铜矿生物堆浸,基于细菌生长Monod方程及收缩核模型建立细菌生长动力学因子影响硫化矿浸出速率的动力学模型,研究铜浸出速率、溶液总铁、溶液中细菌浓度与时间的关系、细菌产出率和细菌饱和系数对浸出速率影响的动力学规律。动力学研究表明,在浸出早期,氧化浸出速率、溶液中总铁浓度以及溶液中的细菌数量增长较快,而在浸出后期则增长较慢。计算与实际结果表明,细菌最大生长比速率、细菌产出率、细菌饱和常数及溶液中Fe离子的浓度均对硫化矿的氧化浸出速率有明显影响,尤其在浸出早期影响较大。应用动力学模型仿真结果与实际基本符合,可分析生物因素对浸出的影响趋势。     

Diversity of microbial community at acid mine drainages from Dachang metals-rich mine, China

Two acid mine drainage (AMD) samples TS and WK, which were from the Dachang metals-rich mine in Guangxi province, China, were studied using PCR-based cloning approach. A total of 44 operational taxonomic units (OTUs) were obtained from the two AMD samples. However, only three OTUs (GXDC-9, GXDC-19 and GXDC-50) detected in sample TS can also be observed in sample WK. Phylogenetic analysis revealed that the bacteria in the two samples fell into four putative divisions, which were Nitrospira, Alphaproteobacteria, Gamaproteobacteria, and Acidobacteria. Organisms of genuses Acidithiobacillus and Leptospirillum, which were in gamaproteobacteria class and Nitrospira family, were dominant in two samples, respectively. In sample TS, which was characterized by low pH, high sulfate, high iron, and high arsenide, two species (Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans) constituted 98.22% of the entire microbial community. Compared with sample TS, the microbial community in sample WK was more diversified according to the observation. Interestedly, the Legionella species, which was rarely observed in the low-pH environment, was detected in sample WK. This work helps us to further understand the diversity of microbial community living in extreme acid mine drainages with unique geochemistry and the tolerance capability of acidophiles to heavy metal.     

Biofilm forming and leaching mechanism during bioleaching chalcopyrite by Thiobacillus ferrooxidans

The mechanism of attachment and leaching of thiobacillus ferrooxidans (T. f. ) on chalcopyrite were studied. The shaking flasks with bacteria were observed by SEM. The process of T. f attached to the surface of the mineral sample and the biofilm forming were described. The promoting role of the biofilm for bioleaching was discussed. The existence of Fe^2 in the exopolysaccharide layer of T. f was demonstrated by EM(electronic microscope)cell-chemistry analysis. These results show that under the proper growth condition of bacteria, bioleaching of chalcopyrite results in the formation of complete biofilm after 2 - 3 weeks. There are iron ions in the outer layer polymer of T. f. , which provides the micro-environment for themselves, and can guaruntee the energy needed for the bacteria growth in the blofilm. At the same time, Fe^3 ions produced oxidize sulfide which brings about the increase of both growth rate of the bacterial and leaching rate of sulfide minerals.     

Microcalorimetric studies of interaction between extracellular polymeric substance and sulfide minerals

Extracellular polymeric substance（EPS） seem to be important for bioleaching. For better control of the processes, the function of EPS of leaching bacteria is of crucial importance. Microcalorimetric measurement was used to study the three simulative components of EPS. The attachment of L-cysteine increases with its increasing concentration, which is Langmuir monomolecular layer adsorption. The attachment of glucose on pyrite is similar to mannose, which is not Langmuir monomolecular layer but multilayer adsorption. The results prove that the EPS mediates attachment and reacts with the sulphide surface and results in changed chemical properties of the mineral surface. It provides a new method to study the interaction between EPS and sulfide minerals.     

海洋环境中油气管道的微生物腐蚀研究进展

海上油气集输管道的腐蚀能够导致严重的环境风险和经济损失，其中微生物腐蚀一直以来被认为是造成该问题的主要因素之一。针对海洋环境油气管网中腐蚀性微生物的来源进行了分类，包括油藏内源性微生物、外注海水以及微生物采油（MEOR）引入的外源性微生物。分析了海底油藏储层中流体化学物质特性，确认其富含甲烷、硫化物、挥发性脂肪酸等，并依据内源微生物代谢及产物特征进行了分类，包括硫酸盐还原菌（SRB）、产甲烷菌、发酵菌以及铁还原菌（IRB）。同时，通过举例分析某油田采出水中微生物群落丰度特征，阐明了外源微生物长期受到油田开采环境胁迫后微生物群落的变化规律。在此基础上，进一步针对海上油气集输管网内涉及的微生物代谢产物理论、电活性微生物腐蚀理论以及腐蚀性微生物之间的协同与拮抗作用进行了全面的归纳总结。最后，对目前以纯培养或模式菌株混合培养为主要方式的微生物腐蚀研究中存在的问题进行了讨论，并对基于生物技术的新型防腐手段进行了展望。     

Elective culture of bacteria used in bioleaching on pyrrhotite

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Development and application of functional gene arrays for microbial community analysis

Functional gene markers can provide important information about fimctional gene diversity and potential activity of microbial communities. Although microarray technology has been successfully applied to study gene expression for pure cultures, simple, and artificial microbial communities, adapting such a technology to analyze complex microbial communities still presents a lot of challenges in terms of design, sample preparation, and data analysis. This work is focused on the development and application of functional gene arrays （FGAs） to larget key functional gene markers for microbial community studies. A few key issues specifically related to FGAs, such as oligonucleotide probe design, nucleic acid extraction and purification, data analysis, specificity, sensitivity, and quantitative capability are discussed in detail. Recent studies have demonstrated that FGAs can provide specific, sensitive, and potentially quantitative information about microbial communities from a variety of natural environments and controlled ecosystems. This technology is expected to revolutionize the analysis of microbial communities, and link microbial structure to ecosystem functioning.     

Single and cooperative bioleaching of sphalerite by two kinds of bacteria —— Acidithiobacillus ferriooxidans and Acidithiobacillus thiooxidans

A cooperative bioleaching （Acidithiobacillus ferriooxidans and Acidithiobacillus thiooxidans） and single bioleaching （Acidithiobacillus ferriooxidans or Acidithiobacillus thiooxidans） of sphalerite were investigated by X-ray diffractometry, energy dispersive spectrography and scanning electron microscopy. The experimental results show that the leaching rate of zinc in the mixed culture is higher than that in pure culture and the sterile control. In these processes, two kinds of bacteria perform different functions and play a cooperative role during leaching of sphalerite. The bioleaching action carded out by Acidithiobacillus ferriooxidans （.4. ferriooxidans） is not directly performed through Fe^2＋ but Fe^3＋, and its role is to oxidize Fe^2＋ to Fe ^3＋ and maintain a high redox potential. Moreover, the addition of an appropriate concentration of ferric iron to the leaching systems is beneficial to zinc dissolution. In the leaching systems without Acidithiobacillus thiooxidans （.A. thiooxidans）, elemental sulfur layers are formed on mineral surface during the dissolution of zinc and block continuous leaching. Acidithiobacillus thiooxidans, however, eliminate the passivation and cause the bioleaching process to continue in the leaching systems. At the same time, protons from the bacterial oxidization of the elemental sulfur layers also accelerate the leaching of zinc.     

重金属污染碱性底泥的自养与异养细菌协同搅拌浸出

探讨了生物浸出法处理湘江河床多种重金属污染的碱性底泥。底泥中的多种重金属对水生生物与人类有较大的毒性以及显著抑制细菌的浸出,利用自养细菌与异养细菌协作的浸出方法可以解决这个问题。结果表明,底泥中锌、锰、铜和镉的生物浸出率分别达95.2%、94.2%、90.1%和84.4%。利用连续提取法分析了浸出前、后底泥中不同形态重金属的含量变化,发现浸出后底泥中残余重金属主要以铁锰氧化态、有机结合态和残渣态的形式存在,这些存在形态的生物毒性较低。本研究说明自养菌结合异养菌的浸出法可有效提高碱性底泥重金属的浸出率和降低生物毒性。     

采用中温菌和常温菌浸出含砷金精矿

考察了不同矿浆浓度和接种量对细菌浸出含砷金精矿的影响。结果表明：中度嗜热嗜酸铁氧化菌MLY和嗜酸氧化亚铁硫杆菌（Acidthiobacillus ferrooxidans,At.f）的浸矿效果不同。MLY和At.f对含砷金精矿的浸出机理也不同;接种MLY的矿浆中,可能主要是溶液中的细菌将Fe^2＋氧化为Fe^3＋,再由Fe^3＋氧化溶解矿物,即以间接浸出作用为主;而接种At.f可能首先是At.f被吸附到矿物表面,通过细菌与矿物之间的某种界面作用引起矿物溶解,矿物溶解产生的Fe^2＋被游离菌氧化成为Fe^3＋,从而溶解矿物,即At.f浸出可能包括直接作用和间接作用的共同效应。