硫酸盐还原菌与油田金属管道腐蚀

硫酸盐还原菌（sulfate-reducing bacteria,SRB）是一类利用硫酸盐和其他氧化态硫化物,或利用元素硫作为电子受体,并将其还原成S2-的原核生物。在长时间开采原油过程中,地层需要通过金属管道注水来维持油气输出压力,而输水管壁上很容易形成生物膜垢。该生物膜垢的形成是微生物腐蚀发生的必要条件,也是微生物对金属腐蚀问题难以解决的主要原因。     

土壤腐蚀网站硫酸盐还原菌的研究

从我国东北、西北、西南和华北的的10多种土壤的苗蚀试验站的钢件周围及腐蚀产物中,分离、纯化了13株硫酸盐还原菌(SRB),测定了它们的形态、生理生化特性及氢化酶活性,据以确定我国广大地区土壤中分布的钢铁晦蚀厌氧腐蚀萄主要为普通脱硫弧菌(Desulfovibriavulgaris)和脱硫脱硫弧菌(D.desulfuricans)。他们对钢的腐蚀速率和其氧化酶活性存在着很好的相关性。     

油田硫酸盐还原菌酸化腐蚀机制及防治研究进展

硫酸盐还原菌(Sulfate reducing bacteria,SRB)是一些厌氧产硫化氢的细菌的统称,是以有机物为养料的厌氧菌。它们广泛分布于pH值6-9的土壤、海水、河水、淤泥、地下管道、油气井、港湾及锈层中,它们生存于好气性硫细菌产生的沉积物下,其最适宜的生长温度是20-30℃,可以在高达50-60℃的温度下生存,与腐蚀相关的最主要的是脱硫脱硫弧菌(Desulfovibrio desulfuricans)。 它们是许多腐蚀问题的主因,例如油田系统金属管路的腐蚀等。在海上油田生产中,海水常被注入油井用于进行2次采油。富含硫酸盐的海水能加速油藏中SRB的生长,随之H₂S大量产生,引起油田水的酸化,H₂S具有毒性和腐蚀性,增加石油和天然气中的硫含量,并可能引起油田堵塞。SRB引起的腐蚀问题是拭待解决的最主要问题。国内外治理该问题的途径主要有物理杀灭、添加化学杀菌剂等方法,但是这些方法成本高,持续效果不显著。近几年来国外学者开始重点关注利用生物竞争排斥技术(Bio-competitive inhibition technology,BCX)控制硫酸盐还原菌的生长代谢的方法,该方法的原理为通过加入特定的药剂,激活油藏中的本源微生物或加入外源微生物,使其与SRB竞争营养源或产生代谢物抑制SRB的生长代谢,进而抑制H₂S的产生。GMT-LATA的科学家对在厌氧油气储层和开采系统中硝酸盐还原菌的作用进行了最早的研究,认为该细菌可以抑制硫酸盐还原菌的代谢活动。随后BCX技术已经在国外部分油田得到了应用,国内还没有在海油生产中应用的报道,但是也有学者对该方法进行了研究。     

腐蚀生物膜垢中硫酸盐还原菌的系统进化分析

从油田回注水系统加压泵后的管道管壁生物膜垢里分离得到的一株嗜热、嗜酸、异养型硫酸盐还原菌, 编号为CW-04。该菌株兼性厌氧、革兰氏阳性、有极生鞭毛、直或弯曲的短杆状,能产孢子, 最佳生长温度47℃, 在20℃~63℃下培养基中均能使测试瓶有黑色沉淀产生。从腐蚀防治的角度研究该菌株的生长曲线。主要生理生化特性包括: 生长pH范围为3.7~10, 最佳生长pH为6~6.5, 最佳生长盐度为0.7%。能够在H2+醋酸盐、甲酸盐、乙酸盐、丙酸盐、乳酸盐、丙酮酸盐和乙醇等电子供体中生长, 也能够利用葡萄糖、果糖和简单的氨基酸作为唯一碳源生长; 能够在硫酸钠、硫代硫酸钠、亚硫酸钠等电子受体中生长, 但是不能利用硝酸盐和硫磺。通过16S rDNA测序以及系列比对分析: 该菌属于细菌界(Bacteria)、厚壁菌门(Firmicutes)、梭菌纲(Clostridia)、梭菌目(Clostridiales)、蛋白胨链球菌科(Peptococcaceae)、脱硫肠状菌属(Desulfotomaculum), 与该属中的种Desulfotomaculum aeronauticumd 相似性达到99%。     

一种肠道硫酸盐还原菌的增菌培养基

目的针对已经分离、纯化的肠道硫酸盐还原菌,建立一种能快速、高效地培养菌体的培养基。方法比较营养丰富的GAM肉汤与常用于培养硫酸盐还原菌的选择性培养基Postgate的培养效果,摸索在GAM肉汤中添加不同浓度的硫酸盐对两种肠道硫酸盐还原菌-Desulfovibrio desulfuricans和Desulfovibrio intestina—zis的培养效果。确定效果最佳的改良GAM培养基配方,并测定在该培养基中D．desulful＇icans的生长曲线。结果与Postgate培养基相比,GAM肉汤能在2d内快速培养D．desulfugicans,但培养至6d时细菌数量大幅降低。在GAM肉汤中添加Na2SO4与FeSO4,在实验浓度范围内,均显著地促进硫酸盐还原菌的生长。在此基础上改良GAM肉汤培养基,培养得到的细菌数量较GAM肉汤显著提高。D．desulfuricans的生长曲线显示,2d时细菌生长达到最高峰,数量可达3．5×10^7 CFU／mL;培养6d,细菌数量为7．3×10^6 CFU／mL。结论基于GAM肉汤改良而得到的增菌培养基,能快速、高效地培养肠道硫酸盐还原菌,为后续进一步研究肠道硫酸盐还原菌的生理功能提供了支持。     

腐蚀生物膜垢中硫酸盐还原菌的系统进化分析

从油田回注水系统加压泵后的管道管壁生物膜垢里分离得到的一株嗜热、嗜酸、异养型硫酸盐还原菌,编号为CW-04.该菌株兼性厌氧、革兰氏阳性、有极生鞭毛、直或弯曲的短杆状,能产孢子,最佳生长温度47℃,在20℃～63℃下培养基中均能使测试瓶有黑色沉淀产生,从腐蚀防治的角度研究该菌株的生长曲线.主要生理生化特性包括:生长pH范围为3.7～10,最佳生长pH为6～6.5,最佳生长盐度为0.7%.能够在H2 醋酸盐、甲酸盐、乙酸盐、丙酸盐、乳酸盐、丙酮酸盐和乙醇等电子供体中生长,也能够利用葡萄糖、果糖和简单的氨基酸作为唯一碳源生长;能够在硫酸钠、硫代硫酸钠、亚硫酸钠等电子受体中生长,但是不能利用硝酸盐和硫磺.通过16S rDNA测序以及系列比对分析:该菌属于细菌界(Bacteria)、厚壁菌门(Firmicutes)、梭菌纲(Clostridia)、梭菌目(Clostridiales)、蛋白胨链球菌科(Peptococcaceae)、脱硫肠状菌属(Desulfotomaculum),与该属中的种Desulfotomaculum aeronauticumd相似性达到99%.     

不同pH值条件下硫酸盐还原菌组成及硫酸盐还原机制分析

【目的】从海洋沉积物中富集获得硫酸盐还原菌群,改变pH值进行培养,分析pH值对硫酸盐还原性质的影响,明确菌群组成和进行硫酸盐还原功能基因预测,探究硫酸盐还原机制。【方法】分析硫酸盐还原菌群在不同pH值条件下的硫酸盐还原率,在此基础上,利用高通量测序技术和PICRUSt软件分析硫酸盐还原菌群优势菌组成及硫酸盐还原相关基因相对丰度。【结果】硫酸盐还原菌群在不同pH值培养条件下的生长和硫酸盐还原率出现显著变化(P<0.01),在pH 5.0时达到峰值,分别为0.34±0.01和96.52%±0.44%。高通量测序数据显示,pH 5.0时菌群丰富度和多样性最高,优势菌属为假单胞菌(Pseudomonas)和芽孢杆菌(Bacillus),相对丰度较高的基因为同化性硫酸盐还原相关基因。【结论】硫酸盐还原菌富集生长的最适pH 5.0,在此条件下的高硫酸盐还原率由同化性硫酸盐还原途径主导,为揭示硫酸盐还原机制提供了实验支持,并拓宽了硫酸盐还原菌实践应用方面的种质资源。     

用硫酸盐还原菌处理重金属废水的研究

介绍了用硫酸盐还原菌处理重金属废水的几种主要方法和原理。硫酸盐还原菌处理含重金属废水主要是通过将可溶性的重金属离子转化成不溶性的金属硫化物、氢氧化物、碳酸盐的方式 ,或直接通过以菌体对重金属离子的吸附完成的。目前研究用硫酸盐还原菌处理重金属废水的主要方法有分批沉淀工艺、吸附处理工艺、化学法和硫酸盐还原菌的混合工艺、全混合处理工艺及硫酸盐还原菌的厌氧上流式污泥床和流化床工艺 ,并对其主要的工艺指标进行了比较。     

硫酸盐还原菌及其还原解毒Cr(Ⅵ)的研究进展

硫酸盐还原菌是一类分布广泛, 能进行硫酸盐异化还原反应的严格厌氧菌。利用硫酸盐还原菌可去除环境中的许多污染物, 因而该类细菌在环境污染治理中具有广阔的应用前景。本文介绍了硫酸盐还原菌的生物学特性和代谢特征及其在环境污染治理中的应用, 并对硫酸盐还原菌还原解毒Cr(Ⅵ)及应用于含Cr(Ⅵ)废水处理的研究进展作了综述, 分析了其未来的研究方向。     

Action of glutaraldehyde and nitrite against sulfate-reducing bacterial biofilms

A continuous flow reactor system was developed to evaluate the efficacy of antimicrobial treatments against sulfate-reducing bacterial biofilms. An annular reactor operating at a nominal dilution rate of 0.5 h⁻¹ was fed one-tenth strength Postgate C medium diluted in 1.5% NaCl and was inoculated with a mixed culture enriched from oilfield-produced water on the same medium. Thin biofilms developed in this reactor after 2 days of operation. The activity of these biofilms resulted in approximately 50 mg S l⁻¹ of sulfide at steady state prior to biocide treatment. Biocide efficacy was quantified by recording the time required for sulfide production to recover following an antimicrobial treatment. In a control experiment in which pure water was applied, the time required to reach 10 mg S l⁻¹ sulfide after the treatment was 1.7±1.2 h, whereas the time to reach this level of sulfide after a pulse dose of 500 mg l⁻¹ glutaraldehyde was delayed to 61±11 h. Nitrite treatment suppressed sulfide production as long as the nitrite concentration remained above 15 mg N l⁻¹. Sulfide production recovered more rapidly after nitrite treatment than it did after glutaraldehyde treatment. Received 01 February 2002/ Accepted in revised form 13 June 2002     

Inhibiting sulfate-reducing bacteria in biofilms by expressing the antimicrobial peptides indolicidin and bactenecin

To identify novel, less-toxic compounds capable of inhibiting sulfate-reducing bacteria (SRB), Desulfovibrio vulgaris and Desulfovibrio gigas in suspension cultures were exposed to several antimicrobial peptides. The bacterial peptide antimicrobials gramicidin S, gramicidin D, and polymyxin B as well as the cationic peptides indolicidin and bactenecin from bovine neutrophils decreased the viability of both SRB by 90% after a 1-h exposure at concentrations of 25–100 μg ml⁻¹. To reduce corrosion by inhibiting SRB in biofilms, the genes for indolicidin and bactenecin were expressed in Bacillus subtilisBE1500 and B. subtilis WB600 under the control of the constitutive alkaline protease (apr) promoter, and the antimicrobials were secreted into the culture medium using the apr signal sequence. Bactenecin was also synthesized and expressed as a fusion to the pro-region of barnase from Bacillus amyloliquefaciens. Concentrated culture supernatants of B. subtilis BE1500 expressing bactenecin at 3 μg ml⁻¹ decreased the viability of Escherichia coli BK6 by 90% and the reference SRB D. vulgaris by 83% in suspension cultures. B. subtilis BE1500 and B. subtilis WB600 expressing bactenecin in biofilms also inhibited the SRB-induced corrosion of 304 stainless steel six to 12-fold in continuous reactors as evidenced by the lack of change in the impedance spectra (resistance polarization) upon addition of SRB and by the reduction in hydrogen sulfide and iron sulfide in batch fermentations with mild steel. A 36-fold decrease in the population of D. vulgaris in a B. subtilis BE1500 biofilm expressing bactenecin was also observed. This is the first report of an antimicrobial produced in a biofilm for in vivo applications and represents the first application of a beneficial, genetically-engineered biofilm for combating corrosion. Received 27 October 1998/ Accepted in revised form 21 February 1999     

硫酸盐还原细菌生物被膜的形成与调控研究进展

硫酸盐还原细菌(sulfate-reducing bacteria,SRB)形成的生物被膜是微生物导致金属锈蚀行为的主要原因,同时也是重金属污水微生物修复技术的关键因子。生物被膜形成及调控机制研究对SRB的防治和利用均十分重要。本文综述了近年来SRB生物被膜的研究进展,包括SRB生物被膜的胞外多聚物组成和控制因子,并着重阐述了目前已知的调控因子对SRB生物被膜形成的影响。     

Planktonic nitrate-reducing bacteria and sulfate-reducing bacteria in some western Canadian oil field waters

Oil fields that use water flooding to enhance oil recovery may become sour because of the production of H₂S from the reduction of sulfate by sulfate-reducing bacteria (SRB). The addition of nitrate to produced waters can stimulate the activities of nitrate-reducing bacteria (NRB) and control sulfide production. Many previous studies have focused on chemolithotrophic bacteria that can use thiosulfate or sulfide as energy sources while reducing nitrate. Little attention has been given to heterotrophic NRB in oil field waters. Three different media were used in this study to enumerate various types of planktonic NRB present in waters from five oil fields in western Canada. The numbers of planktonic SRB and bacteria capable of growth under aerobic conditions were also determined. In general, microbial numbers in the produced waters were very low (<10 ml⁻¹) in samples taken near or at wellheads. However, the numbers increased in the aboveground facilities. No thiosulfate-oxidizing NRB were detected in the oil field waters, but other types of NRB were detected in 16 of 18 produced water samples. The numbers of heterotrophic NRB were equal to or greater than the number of sulfide-oxidizing, chemolithotrophic NRB in 12 of 15 samples. These results showed that each of the oil fields contained NRB, which might be stimulated by nitrate amendment to control H₂S production by SRB. Journal of Industrial Microbiology & Biotechnology (2002) 29, 83–92 doi:10.1038/sj.jim.7000274 Received 20 February 2002/ Accepted in revised form 14 May 2002     

海水养殖生境中硫酸盐还原菌活性的抑制机制

【目的】海水养殖生境中的硫化物(H₂S)严重损害养殖生物健康,控制该条件下硫酸盐还原菌(sulfate-reducing bacteria,SRB)的代谢活性是有效抑制H₂S产生的重要途径。【方法】本研究利用稀释涂布-叠皿夹法对海水养殖生境底泥中SRB进行富集筛选,获得SRB菌株,通过投加硝酸盐对菌株产H₂S的活性进行抑制。【结果】获得的2株SRB Desulfovibrio sp.NY-1和Clostridium sp.NH-1,能够在35℃、pH为7.0及盐度为20–30 mg/L条件下,分别积累高达435和150 mg/L H₂S。硝酸盐不能有效抑制NY-1产H₂S的活性,基因调控作用以及缺乏将硝酸盐作为电子受体的酶体系是其不能被抑制的主要原因。硝酸盐对NH-1 H₂S产生活性有可逆性抑制,其具有硝酸盐异化还原成铵(dissimilatory nitrate reduction to ammonium,DNRA)的能力,优先利用硝酸盐作为电子受体。DNRA作用下的中间代谢产物亚硝酸盐是有效抑制菌株NH-1产H₂S活性的主要原因,其抑制机理主要为抑制菌株的生长繁殖。【结论】硝酸盐对不同SRB菌株具有不同的抑制机制和效果,在进行硫化物污染控制前需要对产生硫化物的SRB菌群进行分析判别。     

Anaerobic oxidation of dimethylsulfide and methanethiol in mangrove sediments is dominated by sulfate-reducing bacteria

The oxidation of dimethylsulfide and methanethiol by sulfate-reducing bacteria (SRB) was investigated in Tanzanian mangrove sediments. The rate of dimethylsulfide and methanethiol accumulation in nonamended sediment slurry (control) incubations was very low while in the presence of the inhibitors tungstate and bromoethanesulfonic acid (BES), the accumulation rates ranged from 0.02–0.34 to 0.2–0.4 nmol g FW sediment⁻¹ h⁻¹, respectively. Degradation rates of methanethiol and dimethylsulfide added were 2–10-fold higher. These results point to a balance of production and degradation. Degradation was inhibited much stronger by tungstate than by BES, which implied that SRB were more important. In addition, a new species of SRB, designated strain SD1, was isolated. The isolate was a short rod able to utilize a narrow range of substrates including dimethylsulfide, methanethiol, pyruvate and butyrate. Strain SD1 oxidized dimethylsulfide and methanethiol to carbon dioxide and hydrogen sulfide with sulfate as the electron acceptor and exhibited a low specific growth rate of 0.010 ± 0.002 h⁻¹, but a high affinity for its substrates. The isolated microorganism could be placed in the genus Desulfosarcina (the most closely related cultured species was Desulfosarcina variabilis , 97% identity). Strain SD1 represents a member of the dimethylsulfide/methanethiol-consuming SRB population in mangrove sediments.     

象山港海域硫酸盐还原菌的时空分布及其影响因素

分别于2007年7月(夏季)、11月(秋季)与2008年1月(冬季)、4月(春季),调查了象山港海域的水样(表层海水和上覆水)及沉积物中的硫酸盐还原菌丰度的时空分布特征及主要影响因素.结果表明,水样及沉积物中硫酸盐还原菌实测值的变化范围为30～2300 cells·ml-1,沉积物中的硫酸盐还原菌数量高于上覆水及表层海水;硫酸盐还原菌丰度的平面分布均呈现不均匀状态,人类开发活动较多的地区明显高于其他站点.根据硫酸盐还原菌数量,对底质进行腐蚀性评价结果表明,象山港大部分范围底质具有中等强度腐蚀性.有机质污染、水温、pH是表层海水中硫酸盐还原菌分布的重要影响因素(P＜0.01);上覆水中的硫酸盐还原菌含量与上覆水的营养盐(NO2--N、NH4+-N)呈显著正相关;沉积物中的硫酸盐还原菌含量与沉积物中的石油类呈显著正相关(P＜0.05).     

硫酸盐还原菌与肠道疾病相关性的研究进展

硫酸盐还原菌(sulfate-reducing bacteria,SRB)是肠道菌群的重要组成成员之一。SRB在其增殖与新陈代谢过程中会产生硫化氢。现有研究显示,SRB的过度繁殖与炎症性肠病(inflammatory bowel disease,IBD)、肠易激综合征(irritable bowel syndrome,IBS)、乳糜泻(celiac disease,CLD)和结直肠癌(colorectal cancer,CRC)等密切相关,但目前尚无相关文献对SRB在肠道疾病中扮演的角色、致病机理和肠道微生态等研究进行系统性的综述。SRB在肠道疾病中的分布特征及其致病机制值得进一步总结与探讨。本文收集过去10年来发表的关于SRB与肠道疾病的文献并进行详细分析与归纳,对SRB在宿主肠道内的分布与生理特征、SRB在不同肠道疾病的相关分布特点,以及SRB致病机制的研究进展等方面进行详细论述,以期增加本领域研究人员对SRB的重视。同时,本文对未来如何深化SRB在肠道相关疾病的研究方向进行探讨,以期为SRB相关肠道疾病的预防与治疗提供一定参考。