硫自养高氯酸盐还原菌和反硝化菌的培养和驯化

为了提高硫自养高氯酸盐和硝酸盐的还原效率,进行了以单质硫(S0)和好氧污泥分别作为电子供体和接种污泥培养驯化硫自养高氯酸盐还原菌和反硝化菌的过程研究.结果显示,在厌氧条件下采用间歇周期运行的培养方式,高氯酸盐和硝酸盐的初始浓度分别从80 mg/L和40 mg/L逐渐增加到240 mg/L和120 mg/L时,去除率达到99%以上的周期随培养驯化时间的增加而逐渐缩短,从最初的7 d稳定在4 d,高氯酸盐和硝酸盐最终分别被还原为Cl-和N2.由于S0的歧化反应,SO42-的实际产生量高于理论产生量.培养驯化过程中pH在6.5-7.3范围内变化,适合硫自养高氯酸盐还原菌和反硝化菌生长、发育和繁殖的条件.PCR-DGGE图谱分析表明随着培养和驯化时间的增加微生物种群数量逐渐减少,污泥微生物群落能够迅速进行优胜劣汰,达到适应环境的目的.培养驯化末期的污泥中β变形杆菌纲处于优势地位,是硫自养高氯酸盐还原菌和反硝化菌的主要组成部分.因此,经培养驯化的微生物菌群具有较高的高氯酸盐和硝酸盐还原能力,本研究可为水中高氯酸盐和硝酸盐净化技术提供一定的理论依据和技术支持.     

生物还原-化学沉淀去除烟气SO2中Na2SO3和硫化物的研究

利用硫酸盐还原菌(SRB)和金属离子沉淀剂对烟气SO2进行生物还原-化学沉淀去除,考察了碳源类型、乙酸(碳源)浓度、沉淀剂投加量和反应时间对SO2-3去除的影响,研究表明,沉淀剂的加入导致SRB转化产生的S2-能很快生成硫化物沉淀,其它副反应被抑制.乙酸C原子/SO2-3的浓度比为1时,SO2-3去除最大.随着沉淀剂投加量的增加,SO2-3去除率增大.ZnCl2生化反应培养14d后,SO2-3去除趋于完全.FeCl2生化反应培养7d后,SO2-3去除趋于完全.     

还原态硫介导沉积物-水系统硝酸盐还原过程研究

沉积物-水系统中存在着硫元素与反硝化(DN)、硝酸盐异化还原成铵(DNRA)过程的耦合。通过向构建的沉积物-水微宇宙系统依次投加0(N组)、50(L组)和75 mg·L^-1(H组)还原态硫,解析各形态氮在上覆水、间隙水和沉积物中的浓度变化与微生物群落结构,以揭示还原态硫对反硝化、硝酸盐异化还原成铵过程的影响。结果显示,还原态硫投加量分别为50和75 mg·L^-1时,系统分别有38.02%和33.27%还原态硫被转化为SO₄^2-。硫氧化作用促进了反硝化的发生,导致L和H组总氮分别比N组减少11%和8%;但是对DNRA过程没有显著影响。此外,具有硫自养反硝化功能的Thiobacillus和Dechloromonas相对丰度之和随着还原态硫浓度增加而上升,N、L和H组两者相对丰度之和分别为5.69%、6.52%和8.70%;而N、L和H组异养硝酸盐还原菌(HNRB)相对丰度分别为0.98%、0.91%和1.05%,其中,DNRA细菌相对丰度均较低(不超过1%),且不受还原态硫浓度的影响。此结果进一步印证了...     

生物还原-化学沉淀去除烟气SO2中Na2 SO3和硫化物的研究

利用硫酸盐还原菌（SRB）和金属离子沉淀剂对烟气SO2进行生物还原-化学沉淀去除,考察了碳源类型、乙酸（碳源）浓度、沉淀剂投加量和反应时间对SO3^2-去除的影响,研究表明,沉淀剂的加入导致SRB转化产生的S^2-能很快生成硫化物沉淀,其它副反应被抑制．乙酸C原子／SO3^2-的浓度比为1时,SO3^2-去除最大．随着沉淀剂投加量的增加,SO3^2-去除率增大．ZnCl2生化反应培养14d后,SO3^2-去除趋于完全．FeCl2生化反应培养7d后,SO3^2-去除趋于完全．     

磁性壳聚糖凝胶微球固定反硝化菌去除地下水中硝酸盐氮

本研究从活性污泥中分离出氢自养反硝化细菌,在厌氧条件下利用氢气作为电子受体,将硝酸盐氮污染物彻底还原为氮气.通过原位共沉淀/柠檬酸钠交联法制备了一种磁性壳聚糖微球,将氢自养反硝化菌固定于磁性壳聚糖微球上组成固定化微生物反硝化体系.利用16SrDNA菌种鉴定、扫描电镜(SEM)、傅里叶红外光谱(FTIR)对固定化前后的材料进行了表征,并与游离的氢自养反硝化菌进行对比,同时进行静态批实验考察了在不同影响因素下硝酸盐去除效果.结果表明,分离出的氢自养反硝化菌属于陶厄氏菌属(MK928401),且被成功固定在磁性壳聚糖微球上;相同时间内,固定化氢自养反硝化菌对硝酸盐氮去除率高出游离细菌59%,说明固定化菌克服了由于游离菌易团聚而限制反硝化速率的缺点;磁性壳聚糖微球的加入,在一定程度上拓宽了氢自养反硝化菌对硝酸盐氮浓度的适应范围,同时拓宽了氢自养反硝化菌对pH的耐受范围;固定化氢自养反硝化菌经5次重复利用后,仍能高效还原硝酸盐氮,相比于游离细菌具有可回收和循环利用性.以上结果得出,以磁性壳聚糖微球固定氢自养反硝化菌,为高效去除地下水中的硝酸盐氮提供了一种更有效的途径.     

1株贫营养好氧反硝化菌的分离鉴定及其脱氮特性

从水库底泥样品中,以硝酸盐为唯一氮源进行驯化、分离筛选出1株能在贫营养及好氧条件下进行高效反硝化的菌株PY8,经过电镜形态学观察、生理生化和16S rDNA序列分析,并基于16SrDNA序列结果,构建了该菌株的系统发育树,最终确定菌株PY8为根瘤菌Rhizobiumsp.。考察了初始pH值、温度、C/N、初始硝酸钠质量浓度、投菌量对菌株PY8硝酸盐还原活性的影响,以及该菌株的异养硝化性能。结果表明,在pH6.0～10.0,温度25～30℃,C/N1.0～9.0,初始硝酸钠质量浓度0.01～0.50g·L-1,投菌量1%～15%时,菌株PY8培养72h后的硝氮去除率可达到95%以上。另外,该菌株具有同时硝化-反硝化作用,在培养过程中氨氮去除率可达到58%左右。实验结果表明,菌株PY8在微污染水体生物脱氮领域中具有很大的应用潜力。     

聚糖菌颗粒污泥的反硝化特性与微生物群落分布

为了解聚糖菌在污泥颗粒化中的脱氮能力及其微生物生态特性,采用反应器工艺、批式试验、显微技术和荧光原位杂交技术来评估其反硝化能力,揭示其微生物群落的微观结构,探索聚糖菌和聚磷菌在不同粒径污泥中的分布特征.结果表明,污泥对有机物的吸收率稳定在90%以上.颗粒污泥的沉降指数(SVI10)稳定在30-50 mL g-1,远低于接种污泥的108.2 mL g-1.聚醣菌颗粒污泥对硝酸盐氮和亚硝酸盐氮的反硝化去除率分别达到了65%和70%,氮气为主要气态产物.聚糖菌颗粒污泥被大量的四联球菌结构所充斥,丝状菌在其中起到了重要的构架和搭桥的作用.荧光原位杂交结果表明聚糖菌可生存于各尺寸颗粒化污泥内;而聚磷菌受到严重抑制只能较少地分布在颗粒污泥的外围空间.上述结果表明,在SBR反应器中采用厌氧搅拌—排水—好氧曝气的处理模式成功培育出具有聚糖特性的颗粒化微生物聚集体,培育成功的颗粒污泥对硝酸盐氮和亚硝酸盐氮均具有良好的反硝化特性.在整体颗粒污泥微生物群落中聚糖菌为优势种群.图8表1参19     

不同C/N值下亚硝酸盐氧化菌和异养菌混合体系的微生物多样性

采用PCR-RFLP技术研究了不同C/N比下亚硝酸盐氧化菌及异养菌混合体系的微牛物多样性,并探讨了微生物菌群结构与其功能(硝化件能)的关系.C/N=0时,混合体系主要由自养菌和寡营养菌(85.1%)组成,包括亚硝酸盐氧化菌(NOB)、拟杆菌门、α-变形菌纲、浮霉菌门和绿色非硫细菌中的一些菌株.C/N=0.44时,混合体系中的自养菌减少,异养菌(主要是γ-变形菌纲的成员)大量出现.C/N=8.82时,γ-变形菌纲的菌株尤其是反硝化菌Pseudomonas sp.占主导(93.8%),与此同时,随着C/N升高,该混合体系的硝化性能也由专一的亚硝酸盐氧化过程转变为同时硝化反硝化过程.微生物菌群结构的转变较好地解释了其硝化性能的改变.本研究揭示了微生物菌群结构与其功能的内在联系,同时表明PCR-RFLP技术与化学分析相结合是研究微生物菌群结构与功能的有力工具.图3表2参13     

滨海含水层咸-淡水过渡带反硝化性能与控制因素研究

滨海含水层受沿海人口规模、工农养殖业发展以及污染物排放等原因的影响,来自陆地的地下水常向沿海输入硝酸盐造成严重的硝酸盐污染问题,反硝化作为去除地下水中硝酸盐污染物的典型反应,探究其对滨海硝酸盐污染防治、生态系统稳定和生物多样性具有重要科学意义。以地下水和含水介质为原料,设计微生物培养试验,利用硝态氮(NO₃^--N)、溶解氧(DO)、酸碱度(p H)和溶解性有机碳(DOC)的变化综合评价研究区反硝化性能,并通过高通量测序揭示微生物培养试验过程中细菌群落结构组成和多样性变化,最后利用响应曲面法探究碳氮比(C/N)、电导率(EC)和DO的交互作用对硝酸盐反硝化性能的影响。结果表明：在微生物培养试验中,硝态氮去除率可达80%以上,p H值维持在7.0左右,DO值维持在6.0-6.5,DOC最大利用率可达65%;高通量测序发现,随着试验的运行,微生物丰富度下降、多样性降低,与反硝化有关的微生物以好氧反硝化菌(假单胞菌属,Pseudomonas;芽胞杆菌属,Bacillus)和有机碳分解菌(福格斯氏菌属,Vogesella)为主;根据响应曲面分析可得,硝...     

铁还原菌介导的氧化铁还原与硝酸盐还原的竞争效应研究

中性厌氧条件下,脱色希瓦氏菌Shewanella decolorationis(S12)能够使用多种电子受体进行厌氧呼吸,包括溶解态的硝酸盐以及难溶态的氧化铁基质。因此,本文通过构建"脱色希瓦氏菌/铁氧化物/硝态氮"的交互反应体系,研究这一体系中铁还原与硝态氮还原的相互作用过程,并对相互作用机制进行初步探讨。结果表明,交互反应体系中氧化铁还原与硝酸盐还原存在明显的竞争关系;随着氧化铁比表面积、可利用态铁含量的增大,硝酸受抑制作用逐渐增强;通过不同温度煅烧而得到的不同结晶度的赤铁矿对硝态氮的抑制作用也被研究。结果还表明,氧化铁结晶度越高,S12对硝态氮的还原抑制作用越小。循环伏安电化学测试结果发现,加入硝酸条件下,铁氧化峰几乎完全消失,表明硝态氮存在条件下,铁还原受到明显抑制。针对以上结果,初步探讨了铁还原与硝态氮还原竞争的原因,即:(1)Fe(Ⅲ)与NO3–同时作为电子受体,因竞争电子而受到抑制;(2)吸附态Fe(Ⅱ)物种还原NO3–,导致Fe(Ⅱ)累积减少,NO3–还原因产生的Fe(Ⅱ)的低速率而被抑制。     

生物硫铁生成菌的选育及其在重金属废水处理中的应用

为开发高效经济、环境友好的重金属废水处理技术,采用亨盖特(Hungate)厌氧技术分离出一株产生物硫铁复合材料的硫酸盐还原菌(Sulfate reducing bacteria,SRB),命名为SRB2,并对该菌株进行了生理特性、培养工艺和废水处理研究.菌体杆状稍有弯曲,革兰氏阴性菌,最佳碳源为乳酸钠.pH范围5.0～9.0,最适pH 7.0,最适温度为35℃,培养3 d生物硫铁量和硫化物含量能达最大值,分别为2.85 g L-1、358.048 mg L-1.正交试验结果表明,乳酸钠9.0 g L-1+组合氮源1.5 g L-1+硫酸亚铁10.0 g L-1时生物硫铁产量最高.16S rDNA序列分析表明,菌株与脱硫弧菌属Desulfovibriodesulfuricans strain 734同源性为99%.采用生物硫铁处理重金属Cu2+、Pb2+、Cd2+废水,结果表明生物硫铁能快速处理含Cu2+、Pb2+、Cd2+废水,2 min去除率达99.8%以上.因此,生物硫铁在各类重金属废水处理与重金属废水污染事故应急处理中具有较好的应用前景.     

养殖水体复合功能菌的分离及其性能

针对养殖水体中因氨态氮、硫化氢和小分子有机酸富营养化引起的污染问题,分离筛选出硝化细菌、反硝化细菌、光合细菌、硫化细菌和生物絮凝菌等具有不同生理功能的污染物治理菌株,经优化配伍制备出性能优良的复合功能菌,结果表明:硝化细菌对氨态氮的去除率达97.8%,亚硝态氮的去除率达95.7%,反硝化细菌对硝态氮的去除率为96.4%,光合细菌和硫化细菌对硫化氢的去除率为55%,微生物絮凝菌的絮凝效率为83%;复合功能菌对CODCr、NH4+-N,总氮、硫化物的去除率分别可达94.3%,89.6%,88.7%和71.3%。     

Metallic wastewater treatment by sulfate reduction using anaerobic rotating biological contactor reactor under high metal loading conditions

An-RBC reactor is highly suited to treat metallic wastewater. Metal removal is due to sulfide precipitation via sulfate reduction by SRB. Cu(II) removal was the best among the different heavy metals. Maximum metal removal is achieved at low metal loading condition. Metal removal matched well with the solubility product values of respective metal sulfide salts. This study was aimed at investigating the performance of anaerobic rotating biological contactor reactor treating synthetic wastewater containing a mixture of heavy metals under sulfate reducing condition. Statistically valid factorial design of experiments was carried out to understand the dynamics of metal removal using this bioreactor system. Copper removal was maximum (>98%), followed by other heavy metals at their respective low inlet concentrations. Metal loading rates less than 3.7 mg/L?h in case of Cu(II); less than 1.69 mg/L?h for Ni(II), Pb(II), Zn(II), Fe(III) and Cd(II) are favorable to the performance of the An-RBC reactor. Removal efficiency of the heavy metals from mixture depended on the metal species and their inlet loading concentrations. Analysis of metal precipitates formed in the sulfidogenic bioreactor by field emission scanning electron microscopy along with energy dispersive X-ray spectroscopy (FESEM-EDX) confirmed metal sulfide precipitation by SRB. All these results clearly revealed that the attached growth biofilm bioreactor is well suited for heavy metal removal from complex mixture.     

一种分离培养硫酸盐还原菌的改进方法

On the base of the characteristics of sulfate-reducing bacteria (SRB) growth and dish cultivated method, a new method for isolation and cultivation of SRB is established. It is dilution spread and repeat dish sandwish cultivated method with the exellence of dilution spread and embeded growth, and it can help separation and identification of SRB. The method has simple operation and less equipment. And using this method, the natural SRB drop was successfully gained, and small agar pieces containing SRB drop were acquired in easiness. The brief SRB transfering operation in strict anaerobic condition was in-deed realized. Fig 1, Ref 7.     

The bacterial symbiont from the hydrothermal vent tubewormRiftia pachyptila is a sulfide specialist

We have investigated the metabolic adaptations of the chemolithotrophic bacterial symbionts ofRiftia pachyptila. Specimens of the tubeworm were collected by submersible from depths of 2600 m at 13°N on the East Pacific Rise in 1987, and 2450 m at the Galápagos Rift in 1988. Isolated bacteria utilize sulfide, but not thiosulfate or sulfite, as their sole reduced-sulfur energy source. The bacteria rapidly oxidize a wide range of sulfide concentrations (5µM to 2 mM), with maximal respiration rates at concentrations >1 mM, and unlike many sulfur-oxidizing bacteria, show no inhibition in oxygen consumption at sulfide concentrations up to 2 mM. Incubations of freshly homogenized trophosome tissue or isolated bacteria with sodium [³⁵S] sulfide and subsequent analysis of sulfur products by high-performance liquid chromatography and flow-through scintillation counting showed that sulfide disappeared almost completely within 1 min. Both soluble and insoluble products of sulfide oxidation were produced. The soluble fraction contained sulfate and polysulfides, with no thiosulfate produced. However, the majority of the radioactivity was in the water-insoluble fraction, mostly as elemental sulfur. Whole-worm experiments under pressure showed a rapid removal of³⁵S-sulfide from the incubation water, with sulfide, sulfate, and polysulfides appearing in the blood within 4 h. There was no utilization of thiosulfate by the whole worms, freshly homogenized trophosome tissue, or isolated bacteria.     

硫酸盐还原菌的生态特性及其应用[综述]

硫酸盐还原菌（ＳＲＢ）是一类形态各异、营养类型多样、能利用硫酸盐或者其他氧化态硫化物作为电子受体来异化有机物质的严格厌养菌．常见属有脱硫弧菌属（Ｄｅｓｕｌｆｏｖｉｂｒｉｏ）,脱硫肠状菌属（Ｄｅｓｕｌｆｏｔｏｍａｃｕｌｕｍ）．因其参与自然界中的多种反应,所以愈来愈得到人们的关注．１　ＳＲＢ的生活环境和条件１．１　ＳＲＢ在环境中的分布［１］自然界中最常见的ＳＲＢ是嗜温的革兰氏阴性、不产芽孢的类型．在淡水及其他含盐量较低的环境中,易分离到革兰氏阳性、产芽孢的菌株．此外,在自然界中存在的还有革兰氏阴性嗜…     

Effect of nitrobenzene on the performance and bacterial community in an expanded granular sludge bed reactor treating high-sulfate organic wastewater

Less than 50 mg/L nitrobenzene brought little effect on anaerobic sulfate reduction. Kinetics of sulfate reduction under different nitrobenzene contents was studied. Increased nitrobenzene contents greatly changed the bacterial community structure. Genus Desulfovibrio played the key role in anaerobic sulfate reduction process. Nitrobenzene (NB) is frequently found in wastewaters containing sulfate and may affect biological sulfate reduction process, but information is limited on the responses of sulfate reduction efficiency and microbial community to the increased NB contents. In this study, a laboratory-scale expanded granular sludge bed reactor was operated continuously to treat high-sulfate organic wastewater with increased NB contents. Results successfully demonstrated that the presence of more than 50 mg/L NB depressed sulfate reduction and such inhibition was partly reversible. Bath experiments showed that the maximum specific desulfuration activity (SDA) decreased from 135.80 mg SO₄^2?/gVSS/d to 30.78 mg SO₄^2?/gVSS/d when the NB contents increased from none to 400 mg/L. High-throughput sequencing showed that NB also greatly affected bacterial community structure. Bacteroidetes dominated in the bioreactor. The abundance of Proteobacteria increased with NB addition while Firmicutes presented an opposite trend. Proteobacteria gradually replaced Firmicutes for the dominance in response to the increase of influent NB concentrations. The genus Desulfovibrio was the dominant sulfate-reducing bacteria (SRB) with absence or presence of NB, but was inhibited under high content of NB. The results provided better understanding for the biological sulfate reduction under NB stress.     

一株好氧反硝化菌的分离鉴定及其混合应用特性研究

采用溴百里酚(BTB)鉴定培养基和稀释平板法从南京市某市政污水处理厂曝气池污水样本中分离筛选得到1株好氧反硝化细菌,经16S rDNA序列同源性比较和系统发育分析初步鉴定为反硝化产碱杆菌(Alcaligenes denitrificans),并将其命名为菌株BMB-N6.研究了菌株BMB-N6在不同浓度亚硝态氮条件下的反硝化能力,运用正交试验设计探讨了该菌株最适的好氧反硝化条件,并且在实验室和大田条件下分别考察了菌株BMB-N6与蛋白质降解菌BMB-LA和氨氮脱除菌BMB-HKF复配形成的混合菌制剂的反硝化能力.结果表明,菌株BMB-N6在8 h内对亚硝态氮的去除率可达94%,其最适亚硝态氮去除条件为摇床转速50 r·min-1,C/N比值4,pH 6,温度35 ℃.在实验室条件下以菌株BMB-N6为基础制成的混合菌制剂在12 h内可去除90%的亚硝态氮,在大田应用中7 d内可去除80%的亚硝态氮.     

Autotrophic denitrification using hydrogen oxidizing bacteria in continuous flow biofilm reactor

Autotrophic denitrification was investigated in five bench‐scale upflow attached growth reactors using hydrogen oxidizing bacteria under anoxic conditions. The performance of sand, granular activated carbon (GAC), crushed pumice, crushed volcanic rock, and plastic media were evaluated as the support material. The reactors were inoculated with acclimated cultures obtained from domestic sewage treatment plant. A synthetic solution containing nitrate was used as the influent. The reactor performance was evaluated by measuring influent and effluent nitrate concentration. The design parameters demonstrated that the effectiveness of autotrophic denitrification is comparable to that of the heterotrophic process and may be utilized economically for drinking water treatment either as the main process or as a supplemental process for ion exchange regenerant treatment.     

Competition for electrons between reductive dechlorination and denitrification

It is common that 2,4,6-trichlorophenol (TCP) coexists with nitrate or nitrite in industrial wastewaters. In this work, simultaneous reductive dechlorination of TCP and denitrification of nitrate or nitrite competed for electron donor, which led to their mutual inhibition. All inhibitions could be relieved to a certain degree by augmenting an organic electron donor, but the impact of the added electron donor was strongest for TCP. For simultaneous reduction of TCP together with nitrate, TCP’s removal rate value increased 75% and 150%, respectively, when added glucose was increased from 0.4 mmol?L^–1 to 0.5 mmol?L^–1 and to 0.76 mmol?L^–1. For comparison, the removal rate for nitrate increased by only 25% and 114% for the same added glucose. The relationship between their initial biodegradation rates versus their initial concentrations could be represented well with the Monod model, which quantified their half-maximum-rate concentration (K _S value), and K _S values for TCP, nitrate, and nitrite were larger with simultaneous reduction than independent reduction. The increases in K _S are further evidence that competition for the electron donor led to mutual inhibition. For bioremediation of wastewater containing TCP and oxidized nitrogen, both reduction reactions should proceed more rapidly if the oxidized nitrogen is nitrite instead of nitrate and if readily biodegradable electron acceptor is augmented.

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