富集培养条件下湖泊和沿海海域水体硫酸盐还原菌的耐氧性特征

硫酸盐还原菌（sulfate-reducing bacteria,SRB） 是一类兼性厌氧菌,在湖泊和海洋有机物矿化过程和生物源性黄铁矿 的生成过程中都扮演着重要角色。环境溶解氧浓度对硫酸盐还原过程影响较大,硫酸盐还原菌在水体中的耐氧性是目前的研 究热点。文章采集了象山港和水口水库不同溶解氧水平下的水样,并在相应的溶解氧梯度下进行富集培养,以探讨不同溶解 氧浓度下硫酸盐还原菌的耐氧性特征及硫代谢相关菌的组成。结果显示,在富集培养条件下湖泊和沿海海域中Desulfovibrio （脱硫弧菌属） 和Desulfomicrobium （脱硫微菌属） 为主要硫酸盐还原细菌,而Shewanella （希瓦氏菌属） 和Sulfurospirillum （硫小螺体属） 为其硫代谢相关菌。Desulfovibrio的相对丰度与溶解氧水平密切相关,随溶解氧浓度的减少,其相对丰度增加。 SRB 的耐氧上限为6.68 mg/L,明显高于以往纯培养或共培养的耐氧上限值。作者推测这不仅与其高氧环境的适应策略有关, 还可能得益于共存菌的贡献,后者可能通过消耗环境中的氧为Desulfovibrio提供生态位,提高其耐氧水平。     

硫酸盐还原菌分离及其脱硫性能的研究

采用单因素及静态实验方法,研究从污水厂活性污泥中富集分离的硫酸盐还原菌(sulfate-reducing bacteria,SRB)在不同初始pH值条件下对SRB脱硫率的影响。试验结果显示,该SRB菌种生长在偏弱酸-中性环境中。当pH=6~7时,最适宜SRB生长,脱硫率达到最高;当pH=3~4时SRB难以生长,脱硫率接近于零。菌液中的金属铀的存在会抑制SRB菌的生长,铀浓度越高,抑制作用越严重。     

江汉平原浅层含水层中土著硫酸盐还原菌对砷迁移释放的影响

硫酸盐还原菌是厌氧环境中参与砷形态转化的重要微生物种群,其介导的生物地球化学循环过程对铁氧化物表面吸附态砷迁移转化的影响亟待深入研究.选取江汉平原典型高砷含水层原位沉积物分离纯化出一株严格厌氧硫酸盐还原菌Desulfovibrio JH-S1,对其进行砷和铁还原能力鉴定,并通过模拟培养实验探究硫酸盐还原菌参与下的铁矿物相转化对吸附态砷迁移的影响.Desulfovibrio JH-S1具有Fe(III)还原能力,无硫和有硫体系中Fe(III)均能被还原,但在硫酸盐充足条件下铁还原量显著增加;该菌株不具备As(V)还原能力,但添加硫酸盐的培养体系中As(V)去除率可达96%以上.Desulfovibrio JH-S1能够还原硫酸盐从而促进载砷的水铁矿还原转化为纤铁矿,并导致吸附的砷释放.江汉平原高砷含水层土著硫酸盐还原菌兼具硫酸盐/铁还原功能,参与了高砷含水层系统中砷-铁-硫耦合循环,对高砷地下水的形成具有重要作用.      

硫酸盐还原菌的驯化培养及脱硫性能研究

采用单因素法及静态实验法研究驯化所得硫酸盐还原菌(sulfate-reducing bacteria，SRB)，旨在对影响菌群生长的因素及脱硫性能进行分析和研究。结果显示，该菌群属于中温菌，培养与应用均以35C为宜；生长环境酸碱度较宽，适宜的初始pH值为7．5左右；可在较高的氯化钠浓度下生存，适宜的浓度范围为10%左右。同时，脱硫实验表明菌的脱硫性能和硫酸根离子的去除率随硫酸根浓度的增加逐渐降低     

海南红树林湿地可培养硫酸盐还原菌的垂直分布特征研究

利用厌氧微生物分离技术,对深度为1.2 m 的海南红树林湿地沉积物钻孔样品进行了分离培养,共获得11 株 厌氧sulfate-reducing bacteria（SRB） 菌株。经显微观察和16S rDNA序列分析,可归纳为6个属,其中已经报道有芽孢杆菌 属（Bacillus）、弧菌属（Vibrio） 和梭状芽胞杆菌属（Clostridium）,另外3个属分别为伯克霍尔德菌属（Burkholderia）、希瓦氏菌属（Shewanella） 和海杆菌属（Marinobacterium）。不同属的细菌对硫酸盐还原的速率最低为14.71%,最高可达 56.78%,并且以上6属11株菌都能将+6价的硫还原生成-2价硫,并与培养基中的Fe2+结合生成黑色FeS沉淀,而这些无定 形FeS沉淀是生成黄铁矿的前体。红树林湿地SRB种群数量随沉积物深度的增加而降低,结合沉积物的地球化学分析测试 结果表明,表层（0 cm） 水界面的沉积物由于处于氧化-还原界面,氧气的周期性输入在一定程度上抑制了SRB的生长;随着 深度增加（10~40 cm）,充足的有机质、偏中性的pH值以及厌氧环境的增强,使得SRB种类和数量明显增加;而60 cm以下 沉积物中因TOC含量降低,减少了微生物可利用的碳源,pH值明显降低,Na+和Ca2+离子浓度明显增加,这些因素都抑制了 SRB的生长,使得深部沉积物中SRB的种类和数量显著减少。     

十红滩铀矿床中微生物种类多样性及生态分布规律的研究

利用生物学方法对新疆十红滩砂岩型铀矿各亚带矿石中参与C、N、S、Fe、O等元素循环的主要微生物菌群进行了研究.结果表明:在铀矿床中赋存着硫酸盐还原菌(SRB)、铁细菌(IB)、硫杆菌、硝化细菌、亚硝化细菌、反硝化细菌、芽孢菌等各类细菌.其中sRB种类较多,主要为脱硫弧菌属、脱硫肠状菌属、脱磺单胞菌属、脱硫杆菌性等.IB主要为鞘铁菌属、球衣菌属、瑙曼氏菌属和赭菌属.硫杆菌包括氧化硫硫杆菌、氧化亚铁硫杆菌、排硫硫杆菌、脱氮硫杆菌.硝化细菌主要为硝化杆菌属.亚硝化细菌主要为亚硝化单孢菌属.异养反硝化细菌为施氏假单胞菌和类鼻疽假单胞菌,自养反硝化细菌为脱氮硫杆菌.芽孢杆菌包括巨大芽孢杆菌、蜡状芽孢杆菌、侧孢芽孢杆菌、蜂房芽孢杆菌、球形芽孢杆菌.实验证明:各类细菌的种类和数量在铀矿床的空间分布上存在着一定的规律性对铀矿床的地球化学分带性具有一定的指示作用.     

层间氧化带砂岩型铀矿床微生物地球化学特征及与铀成矿的关系研究——以吐鲁番-哈密盆地十红滩铀矿床为例

通过样品采集、细菌培养和鉴定等，首次在中国大型的十红滩层间氧化带砂岩型铀矿床容矿层中发现不同种群的细菌。氧化带主要分布铁细菌、硫杆菌、硝化菌等好氧菌，矿石带主要分布厌氧的硫酸盐还原菌，其种类和数量上具明显的生物地球化学分带性，并与岩石的地球化学分带性一致。首次利用容矿层的硫酸盐还原菌在室内进行了硫酸盐还原试验。根据细菌的代谢特征，结合硫酸盐还原菌还原试验，分析了细菌在层间氧化带形成、铀氧化迁移和还原成矿过程中的作用。     

铁氧化物对硫酸盐还原菌分解硫酸盐矿物的协同作用

以牛肉膏为碳源,用活性污泥混合菌接种,探讨在缺氧条件下添加不同的铁氧化物对硫酸盐还原菌(SRB)分解硫酸盐矿物的影响。通过溶液pH、铁离子、硫酸根浓度以及固体产物的SEM和EDS图谱分析,揭示硫酸盐矿物分解过程和机制。实验结果表明,铁氧化物对SRB分解硫酸盐矿物起着明显的协同作用:①被铁还原菌还原的Fe2+与硫酸盐还原产生的硫化氢反应形成铁硫化物,消除硫化氢对SRB分解硫酸盐的抑制作用;②铁氧化物还原溶解,提高体系的pH和碱度,增加生化产物CO2的溶解,诱导溶解的钙离子形成方解石沉淀,促进SRB分解硫酸盐矿物的过程。     

硫酸盐还原菌粘附作用对硬石膏分解的影响

硫酸盐还原菌(SRB)分解硫酸盐矿物对C、S、Fe、Sr、Ba等元素的循环起着重要制约作用,二者相互作用机制的阐明具有重要的矿物学、地球化学及地质微生物学意义。通过设计厌氧实验,本文探讨了SRB与硬石膏的作用过程及机理。结果表明,较之无菌体系,SRB体系中氧化还原电位(EORP)显著降低;可挥发性硫(AVS)与蛋白质浓度则不断增大;硬石膏中总溶出硫含量增加;硬石膏表面SRB粘附位置出现明显溶蚀现象。分析表明SRB通过两种机制促进硬石膏分解:SRB还原代谢消耗溶解态SO42-,降低SO42-浓度,进而促使硬石膏持续溶解;SRB粘附于硬石膏表面,其自身及代谢产物通过络合硬石膏中的Ca加速矿物分解,此种机制在前人的研究中被普遍忽略,而通过此机制SRB亦可促进难溶硫酸盐(天青石、重晶石等)及铁氧化物的溶解,进而制约相关元素的地球化学行为。     

硫酸盐还原菌及其代谢物的接触作用影响石膏分解的实验研究

采用透析的方法,研究了硫酸盐还原菌(SRB)及不同分子量(0~200、200~2 000、2 000~10 000)代谢产物与石膏接触时对其还原分解的影响。实验结果表明,SRB代谢产生了更多的碱性物质,导致体系pH值明显上升;随着SRB及其代谢产物与石膏的接触作用逐渐增强,石膏的还原分解能力逐渐提高;SRB及其代谢产物可以诱导形成具有良好结晶形态的方解石和无定形态碳酸盐沉淀;代谢产物除可促进石膏还原分解外,一定条件下也可抑制石膏溶解。研究结果表明,SRB及其代谢物与石膏的接触作用对其还原分解有一定的影响,影响机制主要为络合作用和营造pH值、HCO-3浓度不同于溶液的SRB-石膏界面微环境。     

Elucidating microbial processes in nitrate- and sulfate-reducing systems using sulfur and oxygen isotope ratios: The example of oil reservoir souring control

Sulfate-reducing bacteria (SRB) are ubiquitous in anoxic environments where they couple the oxidation of organic compounds to the production of hydrogen sulfide. This can be problematic for various industries including oil production where reservoir “souring” (the generation of H₂S) requires corrective actions. Nitrate or nitrite injection into sour oil fields can promote SRB control by stimulating organotrophic nitrate- or nitrite-reducing bacteria (O-NRB) that out-compete SRB for electron donors (biocompetitive exclusion), and/or by lithotrophic nitrate- or nitrite-reducing sulfide oxidizing bacteria (NR-SOB) that remove H₂S directly. Sulfur and oxygen isotope ratios of sulfide and sulfate were monitored in batch cultures and sulfidic bioreactors to evaluate mitigation of SRB activities by nitrate or nitrite injection. Sulfate reduction in batch cultures of Desulfovibrio sp. strain Lac15 indicated typical Rayleigh-type fractionation of sulfur isotopes during bacterial sulfate reduction (BSR) with lactate, whereas oxygen isotope ratios in unreacted sulfate remained constant. Sulfur isotope fractionation in batch cultures of the NR-SOB Thiomicrospira sp. strain CVO was minimal during the oxidation of sulfide to sulfate, which had δ¹⁸O_SO4 values similar to that of the water-oxygen. Treating an up-flow bioreactor with increasing doses of nitrate to eliminate sulfide resulted in changes in sulfur isotope ratios of sulfate and sulfide but very little variation in oxygen isotope ratios of sulfate. These observations were similar to results obtained from SRB-only, but different from those of NR-SOB-only pure culture control experiments. This suggests that biocompetitive exclusion of SRB took place in the nitrate-injected bioreactor. In two replicate bioreactors treated with nitrite, less pronounced sulfur isotope fractionation and a slight decrease in δ¹⁸O_SO4 were observed. This indicated that NR-SOB played a minor role during dosing with low nitrite and that biocompetitive exclusion was the major process. The results demonstrate that stable isotope data can contribute unique information for understanding complex microbial processes in nitrate- and sulfate-reducing systems, and offer important information for the management of H₂S problems in oil reservoirs and elsewhere.     

我国砂岩型铀矿分带特征研究现状及存在问题

作为一种重要的国家战略资源,砂岩型铀矿床是当今世界上最重要的铀矿床类型之一。本文详细地介绍了砂岩型铀矿在国内外的分布特征及占比情况,并对外生地质作用矿床类型中表生流体作用形成的层间渗透砂岩型和潜水渗透砂岩型铀矿床进行了讨论,发现层间渗透砂岩型铀矿床在外表颜色、矿物组合以及地球化学等方面均具有明显的氧化-还原分带现象,此外,矿床内部还具有细菌分带现象。颜色分带在氧化带、氧化-还原过渡带以及还原带之间具有明显不同的特征;矿物组合在不同分带之间各不相同;地球化学分带表现为U、TOC含量以及Fe~(2+)/Fe~(3+)、Th/U比值在各分带之间差异较大。此外,硫酸盐还原菌、硫杆菌、铁细菌及硝化菌等细菌在不同分带之间的数量相差悬殊,而且硫酸盐还原菌数量与TOC呈明显正相关性。通过矿化带内的碳、硫同位素分析,发现硫酸盐还原菌参与了成矿过程,推测其可能是导致碳、硫同位素分馏的主要因素。总体来看,颜色分带、矿物分带、地球化学分带以及细菌分带均与氧化-还原分带呈耦合关系。本文通过总结层间渗透砂岩型和潜水渗透砂岩型铀矿床的成矿模式和当前分带研究中存在的问题,提出了由细菌、地球化学反应参与的砂岩型铀矿床成矿机理,以及未来亟需解决的若干关键科学问题。典型砂岩型铀矿床的分带现象在物、化、探、遥等领域的异常响应对寻找砂岩型铀矿床具有重要的指导意义。     

长江口盐沼硫酸盐还原菌的分布特征与环境机制

选择长江口崇明东滩两类植被区（互花米草、土著植被）纵向剖面,根据不同高程部位柱样土壤和植被根际硫酸盐还原菌（SRB）与异养菌的数量,颗粒有机碳（POC）含量与δ13C值,孔隙水Cl-/SO2-   ４摩尔比等,研究盐沼SRB的空间分布特征与机制。自高潮滩向光滩,柱样氧含量降低,SO2-   ４含量增加,导致SRB含量增加,SRB在有机质矿化中的作用增强。高潮滩柱样不同深度层位的异养菌数量明显大于中潮滩和光滩柱样的相应层位,这与由陆向海柱样氧含量及POC含量降低有关。SRB对植被根际环境的变化较敏感,芦苇根际最有利于SRB生长,藨草根际次之;互花米草根际不利于SRB生长,根系分泌物可能对SRB有抑制作用。土壤有机质含量是导致不同纵向剖面相同高程部位柱样之间微生物数量差异的主要因素。同一纵向剖面不同高程部位柱样之间在土壤氧含量与SO2-   ４含量方面的不同,导致这些柱样微生物数量存在差异。盐沼柱样孔隙水Cl-/SO2-４摩尔比总体偏低,多小于19.33,指示有非海源SO2- ４加入。非海源SO2-４的加入掩盖了硫酸盐还原作用对SO2-４的消耗,增加了利用孔隙水Cl-/SO2-４摩尔比定量研究盐沼硫酸盐还原作用强度的难度。柱样中SRB越多,Cl-/SO2-４摩尔比就越大,硫酸盐还原作用就越明显。     

Kinetic oxygen isotope effects during dissimilatory sulfate reduction: A combined theoretical and experimental approach

Kinetic isotope effects related to the breaking of chemical bonds drive sulfur isotope fractionation during dissimilatory sulfate reduction (DSR), whereas oxygen isotope fractionation during DSR is dominated by exchange between intercellular sulfur intermediates and water. We use a simplified biochemical model for DSR to explore how a kinetic oxygen isotope effect may be expressed. We then explore these relationships in light of evolving sulfur and oxygen isotope compositions (δ³⁴S_SO4 and δ¹⁸O_SO4) during batch culture growth of twelve strains of sulfate-reducing bacteria. Cultured under conditions to optimize growth and with identical δ¹⁸O_H2O and initial δ¹⁸O_SO4, all strains show ³⁴S enrichment, whereas only six strains show significant ¹⁸O enrichment. The remaining six show no (or minimal) change in δ¹⁸O_SO4 over the growth of the bacteria. We use these experimental and theoretical results to address three questions: (i) which sulfur intermediates exchange oxygen isotopes with water, (ii) what is the kinetic oxygen isotope effect related to the reduction of adenosine phosphosulfate (APS) to sulfite (SO₃²⁻), (iii) does a kinetic oxygen isotope effect impact the apparent oxygen isotope equilibrium values? We conclude that oxygen isotope exchange between water and a sulfur intermediate likely occurs downstream of APS and that our data constrain the kinetic oxygen isotope fractionation for the reduction of APS to sulfite to be smaller than 4‰. This small oxygen isotope effect impacts the apparent oxygen isotope equilibrium as controlled by the extent to which APS reduction is rate-limiting.     

阔尔真阔腊浅成低温热液金矿床成矿作用与成矿模式初探

对阔尔真阔腊金矿床的成矿温度、微量元素与稀土元素含量、流体包裹体成份、硫、氢、氧、铷、锶等同位素资料的分析对比 ,探讨了该金矿床的成矿作用及其主要过程。认为该金矿床是一产于地洼区的浅成低温热液矿床 ,具有一系列多因复成矿床的基本成矿特征 ,其成矿至少经历了海西期地槽阶段的低温热液富集及中亚期地洼阶段的活化改造这两个重要阶段 ,并建立了其新的成矿模式。     

Microbially influenced formation of Mg‐calcite and Ca‐dolomite in the presence of exopolymeric substances produced by sulphate‐reducing bacteria

Some species of sulphate‐reducing bacteria (SRB) are known to mediate the formation of dolomite and Mg‐calcite. However, their exact role in the mineralization process remains elusive. Here, we present the result of a laboratory experiment that was designed to test whether formation of carbonate minerals by SRB can occur in the absence of living cells, through passive mineralization of their exopolymeric substances (EPS). SRB capable of mediating dolomite were cultivated in the laboratory, allowing them to secrete EPS. Microbial activity within the cultures was subsequently inhibited with antibiotics. Only after this step, Ca²⁺ and Mg²⁺ were added to the solution and carbonate minerals could form. Mg‐calcite and disordered Ca‐dolomite precipitated in association with EPS. The mol.% of Mg²⁺ in the crystals increased with longer incubation times. This result demonstrates that organic compounds produced by SRB can mediate the formation of Ca‐Mg carbonates in the absence of an active metabolism.     

Stable isotope fractionation during bacterial sulfate reduction is controlled by reoxidation of intermediates

Bacterial sulfate reduction is one of the most important respiration processes in anoxic habitats and is often assessed by analyzing the results of stable isotope fractionation. However, stable isotope fractionation is supposed to be influenced by the reduction rate and other parameters, such as temperature. We studied here the mechanistic basics of observed differences in stable isotope fractionation during bacterial sulfate reduction. Batch experiments with four sulfate-reducing strains (Desulfovibrio desulfuricans, Desulfobacca acetoxidans, Desulfonatronovibrio hydrogenovorans, and strain TRM1) were performed. These microorganisms metabolize different carbon sources (lactate, acetate, formate, and toluene) and showed broad variations in their sulfur isotope enrichment factors. We performed a series of experiments on isotope exchange of ¹⁸O between residual sulfate and ambient water. Batch experiments were conducted with ¹⁸O-enriched (δ¹⁸O_water = +700‰) and depleted water (δ¹⁸O_water = −40‰), respectively, and the stable ¹⁸O isotope shift in the residual sulfate was followed. For Desulfovibrio desulfuricans and Desulfonatronovibrio hydrogenovorans, which are both characterized by low sulfur isotope fractionation (ε_S > −13.2‰), δ¹⁸O values in the remaining sulfate increased by only 50‰ during growth when ¹⁸O-enriched water was used for the growth medium. In contrast, with Desulfobacca acetoxidans and strain TRM1 (ε_S < −22.7‰) the residual sulfate showed an increase of the sulfate δ¹⁸O close to the values of the enriched water of +700‰. In the experiments with δ¹⁸O-depleted water, the oxygen isotope values in the residual sulfate stayed fairly constant for strains Desulfovibrio desulfuricans, Desulfobacca acetoxidans and Desulfonatronovibrio hydrogenovorans. However, strain TRM1, which exhibits the lowest sulfur isotope fractionation factor (ε_S < −38.7‰) showed slightly decreasing δ¹⁸O values.Our results give strong evidence that the oxygen atoms of sulfate exchange with water during sulfate reduction. However, this neither takes place in the sulfate itself nor during formation of APS (adenosine-5′-phosphosulfate), but rather in intermediates of the sulfate reduction pathway. These may in turn be partially reoxidized to form sulfate. This reoxidation leads to an incorporation of oxygen from water into the “recycled” sulfate changing the overall ¹⁸O isotopic composition of the remaining sulfate fraction. Our study shows that such incorporation of ¹⁸O is correlated with the stable isotope enrichment factor for sulfur measured during sulfate reduction. The reoxidation of intermediates of the sulfate reduction pathway does also strongly influence the sulfur stable isotope enrichment factor. This aforesaid reoxidation is probably dependent on the metabolic conversion of the substrate and therefore also influences the stable isotope fractionation factor indirectly in a rate dependent manner. However, this effect is only indirect. The sulfur isotope enrichment factors for the kinetic reactions themselves are probably not rate dependent.