一株中度嗜热嗜酸硫氧化杆菌的分离和系统发育分析

从云南腾冲温泉酸性水样分离得到一株中度嗜热嗜酸硫氧化杆菌MTH 0 4 ,对分离菌株进行了形态、生理生化特性研究及 1 6SrDNA序列分析。该菌株为革兰氏阴性细菌 ,短杆状 ,菌体大小 (0 6～ 0 8) μm× (1～ 2 ) μm ,化能自养 ,可利用硫磺、四硫酸盐、硫代硫酸盐为能源生长 ,不能利用蛋白胨、葡萄糖、酵母粉 ,也不能进行混合型生长。最适生长温度在 4 0℃～ 4 5℃之间 ,最适生长pH 2 0～ 3 0 ,代时 8h。以 1 6SrDNA序列同源性为基础构建了包括 1 3株相关种属在内的系统发育树 ,结果表明 ,MTH 0 4与喜温硫杆菌 (Thiobacilluscaldus)处于同一进化树分支中 ,相似性达 99 5 %以上     

中度嗜热硫氧化细菌的分离及其特性

从我国酸热环境采样,在硫代硫酸钠洋菜平板上通过单菌落形成,分离到四株类似氧化硫硫杆菌（Thiobacillus thiooxidans）的中度嗜热硫氧化细菌,称作 MS（MS252,MS272,MS281,MS301）。MS菌落微小（0．5～1．0mm）半透明,延长培养时间时呈浅褐色。细胞杆状（0．5～0,8X1．0～2．0um）,极生鞭毛,运动,G（一）。所有菌株最适生长温度为45℃;最适生长初始pH为 3．0～3．5,范围为 1．0～5．5（MS252,MS301）T1．6～6．0（MS272,MS281）。MS272DNA中G＋C含量为57mol％,其它三株均为60mol％。它们与中温的氧化硫硫杆菌一样,能利用元素硫、硫代硫酸纳作能源,以二氧化碳为碳源,自养生长产生硫酸。这些茵不能利用有机营养,为专性无机化能自养菌。它们也不能氧化亚铁（FeSO₄）和硫化物如黄铁矿（FeS₂）。二种菌不同之处在于后者生长温度范围较低（10～37℃）,DNA中G＋C含量较低（50～52mol％）。因此本工作分离的MS菌应定为硫杆菌属的一个新种,称作嗜热氧化硫硫杆菌（Thiobacillus thermothiooxidans sp.nov.）。     

一株嗜酸硫氧化菌的分离

从江西德兴分离得到一株嗜酸硫氧化杆菌DX-2,采用双层固体培养基进行分离纯化,对分离菌株进行了形态、生理生化特性研究及16SrRNA序列分析.该菌株为革兰氏阴性细菌,短杆状,菌体大小（0.4～0.5）μm×（1～2）μm,化能自养,可利用硫磺和硫代硫酸盐为能源生长,不能利用亚铁进行生长.以16SrRNA序列同源性为基础构建了相关种属在内的系统发育树,结果表明,DX-2与喜温硫杆菌（Acidithiobacillus caldus）处于同一进化树分支中,相似性达99%以上.考察了不同重金属离子对DX-2的生长的影响.     

浸矿细菌的遗传工程

硫杆菌属(Thiobacillus)的一些种,氧化硫硫杆菌(T.thiooxidans)和氧化亚铁硫杆菌(T.ferrooxidans),广泛分布于硫化矿床的酸性矿水中,最适生长pH为2.0—2.5,是一类革兰氏阴性专性自养细菌。这类细菌广泛应用于有用金属的浸出,特别适合于从低品位的     

不同能源条件下中度嗜热嗜酸细菌多样性分析

采用黄铁矿、黄铜矿、硫酸亚铁和硫粉混合物作为主要能源物质在50°C条件下分别培养中度嗜热细菌混合物,研究其细菌多样性。提取细菌基因组总DNA,采用PCR结合限制性酶切片段多态性分析(RFLP)方法进行细菌16SrRNA基因的系统发育分析,比较不同能源条件下富集培养的混合细菌群落构成的差异。从3个培养物中共获得阳性克隆303个并进行RFLP分析,对29种不同酶切谱型的克隆插入序列进行测定和系统发育分析。大部分序列与已报道的浸矿微生物16SrRNA序列相似性较高(89.1%~99.7%),归属于硫化叶菌属的耐温氧化硫化杆菌(Sulfobacillus thermotolerans)和热氧化硫化杆菌(Sulfobacillus thermosulfidooxidans),嗜酸硫杆菌属的喜温硫杆菌(Acidithiobacillus caldus),钩端螺旋菌属的嗜铁钩端螺旋菌(Leptospirillumferriphilum)以及unculturedforest soil bacterium、uncultured proteobacterium。其中Acidithiobacillus caldus,Sulfobacillus thermotolerans,Leptospirillumferriphilum3种细菌为三类能源物质培养物中的优势细菌类群。L.ferriphilum在黄铁矿培养体系(53.8%)和硫酸亚铁和硫粉为能源的培养体系中(45.9%)中丰度最高;在以黄铜矿为能源物质的培养体系中,S.thermotolerans的比例大幅上升(70.1%)。关键词:中度嗜热细菌;生物多样性;生物浸矿;喜温硫杆菌(Acidithiobacillus caldus);耐温氧化硫化杆菌(Sulfobacillus thermotolerans);嗜铁钩端螺旋菌(Leptospirillumferriphilum)     

一株嗜酸兼性异养菌的分离和系统发育分析

从广东大宝山矿区硫化矿酸性矿坑水中分离获得一株嗜酸兼性异养菌DBS4-1,对该菌株进行了形态、生理生化特性研究及16S rRNA序列分析。分析结果显示:该菌株为革兰氏阴性细菌,球状,菌体直径约为4.0±0.5μm;该菌株兼性异养,具有广泛的底物利用特性,可以利用有机物进行异养生长,同时在自养环境中,还可以利用三价铁、单质硫获得能量进行生长,最适生长温度为30℃左右,最适生长pH约为3.5;该菌16S rRNA序列与Thiobacillus acidophilus(D86511)同源性高达99%。结果表明DBS4-1属于嗜酸硫杆菌属(Acidiphilium sp.),嗜酸种(acidophilum)。     

硫杆菌的分子遗传学研究

硫杆菌（丁litKucjllSS）是一类革兰氏阴性的化能无机营养细菌,也是分布最广,研究得最多,经济意义很大的硫氧化细菌。该属中的氧化硫硫杆菌（T．th。whns）和氧化亚铁流杆菌（T．fer－rtx）xlua叫是极端嗜酸性的专性自养细菌,最适因为2,0～3．5,广泛分布于流化矿床的酸性废水中。这类细菌很早就被广泛应用于贵重金属的浸出和回收,特别适合于从低品位的矿石中浸出稀有资重金属,主要是通过氧化FeSO和Fe＆产生Fe”氧化剂,氧化金属硫化物使之变成可溶性的硫酸盐形式,从而回收有用金属。这种方法具有低成本,低能耗,无污染等特点…     

一株中度嗜热嗜酸细菌的分离与分子鉴定

用稀释分离法,从云南某温泉分离得到一株中度嗜热嗜酸细菌YN06。该菌株短杆状,革兰氏阴性,菌体大小为(0．3-0．5)μm x(1-2．2)μm。16S rDNA和16S—23S间隔区序列分析表明,YN06与嗜酸硫杆菌属的喜温硫杆菌处于同一进化树分支中,相似性均高达99%以上,因此该菌株被鉴定为喜温硫杆菌。     

用于盐碱土壤处理的氧化硫硫杆菌的选育及鉴定

目的:基于氧化硫硫杆菌能够氧化硫产生硫酸的特性,在盐碱土壤治理与改良方面有良好的应用前景.方法:筛选出氧化硫硫杆菌,其代表菌株TT03是细小杆状细菌,依据依据<伯杰氏细菌鉴定手册>第八版中分类,结合16S rDNA序列测定结果.结果:鉴定为嗜酸性氧化硫硫杆菌A.T(Acidithiobacillus thiooxidans).     

污泥中重金属的生物淋滤

生物淋滤法(Bioleaching)是指利用自然界中一些微生物(硫细菌)的直接作用或其代谢产物的间接作用,产生氧化、还原、络合、吸附或溶解反应,将固相中某些不溶性成分(如重金属、硫及其他金属)分离浸提出来的技术.在生物淋滤中,嗜酸性氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans,A.f)和嗜酸性氧化硫硫杆菌(Acidithiobacillus thiooxidans,A.f)被用作有效的淋滤载体[1].这种嗜酸性的化能自养型细菌以大气中的CO2为碳源,以无机物铁或硫为能源来维持生长,不需要提供外来的碳源和电子供体.另外,由于pH值很低,抑制了其他细菌的生长,所以在实际的操作过程中不需要严格的无菌条件.氧化亚铁硫杆菌和氧化硫硫杆菌去除重金属适宜于污水处理厂的开放系统,采用土著嗜酸性氧化亚铁硫杆菌(A.f)和氧化硫硫杆菌(A.f)进行重金属去除.也就是说,处理什么地方的污泥,就在什么地方分离A.f和A.t,这样分离的微生物在生物淋滤过程中能发挥较好的作用.这也是微生物在自然界生长繁殖的特点之一. 本期介绍了王聪、宋存江等[2]从剩余活性污泥中分离得到两株土著硫杆菌,对两株菌进行了分类鉴定,确定二者分别为嗜酸性氧化亚铁硫杆菌杆(Acidithiobacillus ferrooxidans,A.f)和嗜酸性氧化硫硫杆菌(Acidithiobacillus thiooxidans,A.t),将二者的单菌和混合菌分别接种于剩余活性污泥中,进行了为期9 d的生物淋滤,对淋滤过程中的pH变化、氧化还原电位(ORP)以及重金属含量进行了检测.结果表明,生物淋滤9 d的混合菌对于As、Cr、Cu、Ni和Zn的去除效果最好,去除率分别达到了96.09%、93.47%、98.32%、97.88%和98.60%.混合菌生物淋滤对于Cd和Pb的去除率在第6天之后迅速下降,但是A.t单菌淋滤保持较高的去除率,此结果为进一步的应用打下了良好的基础.     

Isolation and characterization of a sulfur-oxidizing chemolithotroph growing on crude oil under anaerobic conditions

Molecular approaches have shown that a group of bacteria (called cluster 1 bacteria) affiliated with the epsilon subclass of the class Proteobacteria constituted major populations in underground crude-oil storage cavities. In order to unveil their physiology and ecological niche, this study isolated bacterial strains (exemplified by strain YK-1) affiliated with the cluster 1 bacteria from an oil storage cavity at Kuji in Iwate, Japan. 16S rRNA gene sequence analysis indicated that its closest relative was Thiomicrospira denitrificans (90% identity). Growth experiments under anaerobic conditions showed that strain YK-1 was a sulfur-oxidizing obligate chemolithotroph utilizing sulfide, elemental sulfur, thiosulfate, and hydrogen as electron donors and nitrate as an electron acceptor. Oxygen also supported its growth only under microaerobic conditions. Strain YK-1 could not grow on nitrite, and nitrite was the final product of nitrate reduction. Neither sugars, organic acids (including acetate), nor hydrocarbons could serve as carbon and energy sources. A typical stoichiometry of its energy metabolism followed an equation: S(2-) + 4NO(3)(-) --> SO(4)(2-) + 4NO(2)(-) (Delta G(0) = -534 kJ mol(-1)). In a difference from other anaerobic sulfur-oxidizing bacteria, this bacterium was sensitive to NaCl; growth in medium containing more than 1% NaCl was negligible. When YK-1 was grown anaerobically in a sulfur-depleted inorganic medium overlaid with crude oil, sulfate was produced, corresponding to its growth. On the contrary, YK-1 could not utilize crude oil as a carbon source. These results suggest that the cluster 1 bacteria yielded energy for growth in oil storage cavities by oxidizing petroleum sulfur compounds. Based on its physiology, ecological interactions with other members of the groundwater community are discussed.     

Isolation of a strain of <Emphasis Type="Italic">Acidithiobacillus caldus</Emphasis> and its role in bioleaching of chalcopyrite

A moderately thermophilic and acidophilic sulfur-oxidizing bacterium named S₂, was isolated from coal heap drainage. The bacterium was motile, Gram-negative, rod-shaped, measured 0.4 to 0.6 by 1 to 2 μm, and grew optimally at 42–45°C and an initial pH of 2.5. The strain S₂ grew autotrophically by using elemental sulfur, sodium thiosulfate and potassium tetrathionate as energy sources. The strain did not use organic matter and inorganic minerals including ferrous sulfate, pyrite and chalcopyrite as energy sources. The morphological, biochemical, physiological characterization and analysis based on 16S rRNA gene sequence indicated that the strain S₂ is most closely related to Acidithiobacillus caldus (>99% similarity in gene sequence). The combination of the strain S₂ with Leptospirillum ferriphilum or Acidithiobacillus ferrooxidans in chalcopyrite bioleaching improved the copper-leaching efficiency. Scanning electron microscope (SEM) analysis revealed that the chalcopyrite surface in a mixed culture of Leptospirillum ferriphilum and Acidithiobacillus caldus was heavily etched. The energy dispersive X-ray (EDX) analysis indicated that Acidithiobacillus caldus has the potential role to enhance the recovery of copper from chalcopyrite by oxidizing the sulfur formed during the bioleaching progress.     

Molecular and morphological characterization of the association between bacterial endosymbionts and the marine nematode Astomonema sp. from the Bahamas

Marine nematode worms without a mouth or functional gut are found worldwide in intertidal sandflats, deep-sea muds and methane-rich pock marks, and morphological studies show that they are associated with endosymbiotic bacteria. While it has been hypothesized that the symbionts are chemoautotrophic sulfur oxidizers, to date nothing is known about the phylogeny or function of endosymbionts from marine nematodes. In this study, we characterized the association between bacterial endosymbionts and the marine nematode Astomonema sp. from coral reef sediments in the Bahamas. Phylogenetic analysis of the host based on its 18S rRNA gene showed that Astomonema sp. is most closely related to non-symbiotic nematodes of the families Linhomoeidae and Axonolaimidae and is not closely related to marine stilbonematinid nematodes with ectosymbiotic sulfur-oxidizing bacteria. In contrast, phylogenetic analyses of the symbionts of Astomonema sp. using comparative 16S rRNA gene sequence analysis revealed that these are closely related to the stilbonematinid ectosymbionts (95-96% sequence similarity) as well as to the sulfur-oxidizing endosymbionts from gutless marine oligochaetes. The closest free-living relatives of these gammaproteobacterial symbionts are sulfur-oxidizing bacteria from the family Chromatiaceae. Transmission electron microscopy and fluorescence in situ hybridization showed that the bacterial symbionts completely fill the gut lumen of Astomonema sp., suggesting that these are their main source of nutrition. The close phylogenetic relationship of the Astomonema sp. symbionts to known sulfur-oxidizing bacteria as well as the presence of the aprA gene, typically found in sulfur-oxidizing bacteria, indicates that the Astomonema sp. symbionts use reduced sulfur compounds as an energy source to provide their hosts with nutrition.     

冲绳海槽热液区可培养硫氧化细菌多样性及其硫氧化特性

冲绳海槽热液区独特的地质环境孕育了特殊的生物群落,硫氧化细菌作为生物地球化学循环的重要参与者在热液生态系统中发挥着至关重要的作用。【目的】通过硫氧化菌株的分离培养揭示冲绳海槽热液区可培养硫氧化细菌的多样性和硫氧化活性。【方法】采用多种培养基对冲绳海槽热液区不同沉积物样品中的硫氧化细菌进行富集培养和分离纯化;利用16S rRNA基因序列确定硫氧化细菌的分类地位并进行系统发育分析;采用碘量法对典型硫氧化菌株硫氧化活性进行检测。【结果】本研究从冲绳海槽热液区样品中共分离鉴定85株硫氧化细菌,分属于α-变形菌纲、γ-变形菌纲、放线菌门和厚壁菌门,优势属为氢弧菌属(Hydrogenovibrio)、拉布伦氏菌属(Labrenzia)、深海海旋菌属(Thalassospira)和海杆状菌属(Marinobacter)。硫氧化活性检测结果表明,7株典型硫氧化菌株对硫代硫酸钠的降解活性介于31%–100%之间,其中泰坦尼克号盐单胞菌SOB56 (Halomonas titanicae SOB56)、南极海杆状菌SOB93(Marinobacter antarcticus SOB93)、印度硫氧化粗杆菌SOB107 (Thioclava indica SOB107)和嗜温氢弧菌CJG136 (Hydrogenovibrio thermophiles CJG136)可以完全降解硫代硫酸钠。【结论】冲绳海槽热液区可培养硫氧化细菌的多样性丰富,为研究该热液区的硫循环过程提供了实验材料和理论基础,多种硫氧化活性菌株的获得极大地丰富了菌种资源,为探究深海热液区硫循环的能量代谢途径和分子机制奠定基础。     

Isolation, characterization, and in situ detection of a novel chemolithoautotrophic sulfur-oxidizing bacterium in wastewater biofilms growing under microaerophilic conditions

We successfully isolated a novel aerobic chemolithotrophic sulfur-oxidizing bacterium, designated strain SO07, from wastewater biofilms growing under microaerophilic conditions. For isolation, the use of elemental sulfur (S(0)), which is the most abundant sulfur pool in the wastewater biofilms, as the electron donor was an effective measure to establish an enrichment culture of strain SO07 and further isolation. 16S rRNA gene sequence analysis revealed that newly isolated strain SO07 was affiliated with members of the genus Halothiobacillus, but it was only distantly related to previously isolated species (89% identity). Strain SO07 oxidized elemental sulfur, thiosulfate, and sulfide to sulfate under oxic conditions. Strain SO07 could not grow on nitrate. Organic carbons, including acetate, propionate, and formate, could not serve as carbon and energy sources. Unlike other aerobic sulfur-oxidizing bacteria, this bacterium was sensitive to NaCl; growth in medium containing more than 150 mM was negligible. In situ hybridization combined with confocal laser scanning microscopy revealed that a number of rod-shaped cells hybridized with a probe specific for strain SO07 were mainly present in the oxic biofilm strata (ca. 0 to 100 micro m) and that they often coexisted with sulfate-reducing bacteria in this zone. These results demonstrated that strain SO07 was one of the important sulfur-oxidizing populations involved in the sulfur cycle occurring in the wastewater biofilm and was primarily responsible for the oxidation of H(2)S and S(0) to SO(4)(2-) under oxic conditions.     

Physiological and Genomic Features of a Novel Sulfur-Oxidizing Gammaproteobacterium Belonging to a Previously Uncultivated Symbiotic Lineage Isolated from a Hydrothermal Vent

Strain Hiromi 1, a sulfur-oxidizing gammaproteobacterium was isolated from a hydrothermal vent chimney in the Okinawa Trough and represents a novel genus that may include a phylogenetic group found as endosymbionts of deep-sea gastropods. The SSU rRNA gene sequence similarity between strain Hiromi 1 and the gastropod endosymbionts was approximately 97%. The strain was shown to grow both chemolithoautotrophically and chemolithoheterotrophically with an energy metabolism of sulfur oxidation and O₂ or nitrate reduction. Under chemolithoheterotrophic growth conditions, the strain utilized organic acids and proteinaceous compounds as the carbon and/or nitrogen sources but not the energy source. Various sugars did not support growth as a sole carbon source. The observation of chemolithoheterotrophy in this strain is in line with metagenomic analyses of endosymbionts suggesting the occurrence of chemolithoheterotrophy in gammaproteobacterial symbionts. Chemolithoheterotrophy and the presence of homologous genes for virulence- and quorum sensing-related functions suggest that the sulfur-oxidizing chomolithotrophic microbes seek animal bodies and microbial biofilm formation to obtain supplemental organic carbons in hydrothermal ecosystems.     

Microbial thiocyanate utilization under highly alkaline conditions

Three kinds of alkaliphilic bacteria able to utilize thiocyanate (CNS-) at pH 10 were found in highly alkaline soda lake sediments and soda soils. The first group included obligate heterotrophs that utilized thiocyanate as a nitrogen source while growing at pH 10 with acetate as carbon and energy sources. Most of the heterotrophic strains were able to oxidize sulfide and thiosulfate to tetrathionate. The second group included obligately autotrophic sulfur-oxidizing alkaliphiles which utilized thiocyanate nitrogen during growth with thiosulfate as the energy source. Genetic analysis demonstrated that both the heterotrophic and autotrophic alkaliphiles that utilized thiocyanate as a nitrogen source were related to the previously described sulfur-oxidizing alkaliphiles belonging to the gamma subdivision of the division Proteobacteria (the Halomonas group for the heterotrophs and the genus Thioalkalivibrio for autotrophs). The third group included obligately autotrophic sulfur-oxidizing alkaliphilic bacteria able to utilize thiocyanate as a sole source of energy. These bacteria could be enriched on mineral medium with thiocyanate at pH 10. Growth with thiocyanate was usually much slower than growth with thiosulfate, although the biomass yield on thiocyanate was higher. Of the four strains isolated, the three vibrio-shaped strains were genetically closely related to the previously described sulfur-oxidizing alkaliphiles belonging to the genus Thioalkalivibrio. The rod-shaped isolate differed from the other isolates by its ability to accumulate large amounts of elemental sulfur inside its cells and by its ability to oxidize carbon disulfide. Despite its low DNA homology with and substantial phenotypic differences from the vibrio-shaped strains, this isolate also belonged to the genus Thioalkalivibrio according to a phylogenetic analysis. The heterotrophic and autotrophic alkaliphiles that grew with thiocyanate as an N source possessed a relatively high level of cyanase activity which converted cyanate (CNO-) to ammonia and CO2. On the other hand, cyanase activity either was absent or was present at very low levels in the autotrophic strains grown on thiocyanate as the sole energy and N source. As a result, large amounts of cyanate were found to accumulate in the media during utilization of thiocyanate at pH 10 in batch and thiocyanate-limited continuous cultures. This is a first direct proof of a "cyanate pathway" in pure cultures of thiocyanate-degrading bacteria. Since it is relatively stable under alkaline conditions, cyanate is likely to play a role as an N buffer that keeps the alkaliphilic bacteria safe from inhibition by free ammonia, which otherwise would reach toxic levels during dissimilatory degradation of thiocyanate.     

Phylogeny and distribution of the soxB gene among thiosulfate-oxidizing bacteria

A PCR protocol for the detection of sulfur-oxidizing bacteria based on soxB genes that are essential for thiosulfate oxidation by sulfur-oxidizing bacteria of various phylogenetic groups which use the 'Paracoccus sulfur oxidation' pathway was developed. Five degenerate primers were used to specifically amplify fragments of soxB genes from different sulfur-oxidizing bacteria previously shown to oxidize thiosulfate. The PCR yielded a soxB fragment of approximately 1000 bp from most of the bacteria. Amino acid and nucleotide sequences of soxB from reference strains as well as from new isolates and environmental DNA from a hydrothermal vent habitat in the North Fiji Basin were compared and used to infer relationships of soxB between sulfur-oxidizing bacteria belonging to various 16S rDNA-based phylogenetic groups. Major phylogenetic lines derived from 16S rDNA were confirmed by soxB phylogeny. Thiosulfate-oxidizing green sulfur bacteria formed a coherent group by their soxB sequences. Likewise, clearly separated branches demonstrated the distant relationship of representatives of alpha-, beta-, and gamma-Proteobacteria including representative species of the former genus Thiobacillus (now Halothiobacillus - gamma-Proteobacteria, Thiobacillus - beta-Proteobacteria and Starkeya - alpha-Proteobacteria). This general picture emerged although apparent evidence for lateral transfer of the soxB gene is indicated and comparison of soxB phylogeny and 16S rDNA phylogeny points to the significance of this gene transfer in hydrothermal vent bacterial communities of the North Fiji Basin.     

Isolation, Characterization, and In Situ Detection of a Novel Chemolithoautotrophic Sulfur-Oxidizing Bacterium in Wastewater Biofilms Growing under Microaerophilic Conditions

We successfully isolated a novel aerobic chemolithotrophic sulfur-oxidizing bacterium, designated strain SO07, from wastewater biofilms growing under microaerophilic conditions. For isolation, the use of elemental sulfur (S⁰), which is the most abundant sulfur pool in the wastewater biofilms, as the electron donor was an effective measure to establish an enrichment culture of strain SO07 and further isolation. 16S rRNA gene sequence analysis revealed that newly isolated strain SO07 was affiliated with members of the genus Halothiobacillus, but it was only distantly related to previously isolated species (89% identity). Strain SO07 oxidized elemental sulfur, thiosulfate, and sulfide to sulfate under oxic conditions. Strain SO07 could not grow on nitrate. Organic carbons, including acetate, propionate, and formate, could not serve as carbon and energy sources. Unlike other aerobic sulfur-oxidizing bacteria, this bacterium was sensitive to NaCl; growth in medium containing more than 150 mM was negligible. In situ hybridization combined with confocal laser scanning microscopy revealed that a number of rod-shaped cells hybridized with a probe specific for strain SO07 were mainly present in the oxic biofilm strata (ca. 0 to 100 μm) and that they often coexisted with sulfate-reducing bacteria in this zone. These results demonstrated that strain SO07 was one of the important sulfur-oxidizing populations involved in the sulfur cycle occurring in the wastewater biofilm and was primarily responsible for the oxidation of H₂S and S⁰ to SO₄²⁻ under oxic conditions.     

Isolation and Characterization of Iron and Sulfur-Oxidizing Bacteria from Maiduk Copper Mine at Shahrbabk Province in Iran

Microbial oxidation of iron and sulfur are important steps in biogeochemical cycles in mining environments. The aim of this study was the enrichment and identification of two important groups of bacteria that are involved in bioleaching of copper ores. Some soil samples were collected from the Maiduk copper mine. Iron-oxidizing bacteria were enriched in 9K medium containing ferrous sulfate, and sulfur oxidizers were enriched in 9K medium containing powdered sulfur instead of ferrous sulfate as energy source. After three subcultures, autotrophic bacteria were isolated on 9K agarose medium with appropriate energy sources. The autotrophic bacteria from the enrichments were identified by amplification of 16S rRNA gene and sequencing. Bioleaching experiments were performed in 100 ml of 9K medium containing 5 g of low-grade copper ore instead of ferrous sulfate. Twelve different iron and sulfur-oxidizing bacteria were isolated from the collected soil samples of Maiduk copper mine. Molecular identification indicated that two prevalent strains in the ore enrichments could be assigned to the Acidithiobacillus ferooxidans strain HGM and the Thiobacillus thioparus strain HGE. These two strains reached their logarithmic phase of growth after 8 days of incubation in their respective media at 30°C. Of these two cultures, strain HGM leached more copper ore (300 ppm) from the Maiduk copper ore than did strain HGE (200 ppm). Application of these two strains to the Maiduk copper ore in situ and to ore heaps should improve the leaching process.