pH值对亚硝酸盐氧化菌动力学及功能基因的影响

为探究pH值对亚硝酸盐氧化菌（NOB）活性动力学影响,本试验采用序批式活性污泥（SBR）反应器,以富含NOB的活性污泥为对象,基于Monod模型考察不同pH值对NOB活性动力学的影响并进行统计学分析.结果表明,Monod方程可较好地反映不同pH值条件下基质底物浓度对NOB比亚硝态氮氧化速率（SNiOR）的影响,且pH=7.0时动力学参数Ks为（6.167mg/L）,r_max为[1.134g/（g·d）],此时NOB活性最好.利用钟形经验模型进行非线性回归拟合,最大比降解速率（r_max）随pH值的增大呈钟形变化,本试验NOB的最佳pH值为（6.9±0.1）,其中r_max维持在r_opt一半以上的pH值范围（ω）为（3.26±0.4）.以亚硝酸盐氧化还原酶类基因（nxrA、nxrB）为引物,基于荧光定量PCR技术分析结果显示,在不同pH值条件下nxrA基因和nxrB基因拷贝数的变化趋势均与动力学参数（Ks、r_max）的规律一致,且nxrA和nxrB基因在系统的降解过程中起协同作用.     

侧流FNA抑制的CANON颗粒-絮体复合系统参数优化

为探究侧流游离亚硝酸(FNA)处理抑制亚硝酸盐氧化菌(NOB)策略相关工艺参数的最佳组合,在颗粒-絮体复合系统的全程自养脱氮(CANON)工艺中,采用批次试验探讨污泥沉降时间、FNA处理浓度及处理时间对相关功能菌活性的影响,针对性抑制NOB活性,降低FNA对功能菌活性影响。结果显示,沉降时间影响处理污泥中功能菌群活性,随着沉降时间的延长,排出污泥中厌氧氨氧化菌(AnAOB)活性逐渐减弱。沉降1min后进行排泥排出了大部分NOB并且反应器内保留了尽可能多的AnAOB,此时排出的污泥中AnAOB、氨氧化菌(AOB)和NOB的相对活性分别为15.79%、54.55%和68.63%。综合FNA对NOB和AOB活性的影响,采用0.6mg/L的FNA抑制12h后AOB活性为38.71%,而NOB活性仅为12.5%。响应曲面分析结果显示,FNA处理时间、处理浓度是影响NOB、AOB活性的关键因素。     

基于不同污泥量间歇饥饿的CANON工艺启动

为了探究不同污泥量间歇饥饿对于全程自养脱氮（CANON）工艺启动的影响,R1、R2和R3反应器分别采用水力筛分间歇饥饿、部分污泥间歇饥饿和全部污泥间歇饥饿3种不同的方式启动CANON工艺.结果显示,R1、R2和R3反应器分别在第44,66和58d顺利启动,第70d时,其总氮去除率分别达到73.63%、71.56%和67.40%左右,粒径分别达到了404,359和306μm.分析表明,水力筛分间歇饥饿方式可以选择性地针对絮状污泥进行饥饿,避免间歇饥饿对于污泥EPS及粒径的负面影响,在有效抑制NOB菌活性的同时易于保持相关功能菌的活性;部分污泥间歇饥饿的方式在一定程度上避免了AOB和ANAMMOX活性的衰减,但同时NOB的抑制效果也较差;整体间歇饥饿策略对于NOB的抑制效果最好,但同时也不利于CANON工艺相关功能菌的活性保持.     

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     

Differences in nitrite-oxidizing communities and kinetics in a brackish environment after enrichment at low and high nitrite concentrations

Nitrite accumulation in shrimp ponds can pose serious adverse effects to shrimp production and the environment.This study aims to develop an effective process for the enrichment of ready-to-use nitrite-oxidizing bacteria(NOB)inocula that would be appropriate for nitrite removal in brackish shrimp ponds.To achieve this objective,the effects of nitrite concentrations on NOB communities and nitrite oxidation kinetics in a brackish environment were investigated.Moving-bed biofilm sequencing batch reactors and continuous moving-bed biofilm reactors were used for the enrichment of NOB at various nitrite concentrations,using sediment from brackish shrimp ponds as seed inoculum.The results from NOB population analysis with quantitative polymerase chain reaction(q PCR)show that only Nitrospira were detected in the sediment from the shrimp ponds.After the enrichment,both Nitrospira and Nitrobacter coexisted in the reactors controlling effluent nitrite at 0.1 and 0.5 mg-NO_2~--N/L.On the other hand,in the reactors controlling effluent nitrite at 3,20,and 100 mg-NO_2~--N/L,Nitrobacter outcompeted Nitrospira in many orders of magnitude.The half saturation coefficients(Ks)for nitrite oxidation of the enrichments at low nitrite concentrations(0.1 and 0.5 mg-NO_2~--N/L)were in the range of 0.71–0.98 mg-NO_2~--N/L.In contrast,the Ksvalues of NOB enriched at high nitrite concentrations(3,20,and 100 mg-NO_2~--N/L)were much higher(8.36–12.20 mg-NO_2~--N/L).The results suggest that the selection of nitrite concentrations for the enrichment of NOB inocula can significantly influence NOB populations and kinetics,which could affect the effectiveness of their applications in brackish shrimp ponds.     

Achieving nitritation at low temperatures using free ammonia inhibition on Nitrobacter and real-time control in an SBR treating landfill leachate

Free ammonia(FA) inhibition on nitrite-oxidized bacteria(NOB) and real-time control are used to achieve nitrogen removal from landfill leachate via nitrite pathway at low temperatures in sequencing batch reactor. The inhibition of FA on NOB activity during the aerobic period was prolonged using real-time control. The degree of nitrite accumulation was monitored along with variations of the ammonia-oxidizing bacteria and NOB population using fluorescence in situ hybridization techniques. It is demonstrated that the end-point of ammonia oxidization is detected from the on-line measured dissolved oxygen,oxidization–reduction potential, and p H signals, which could avoid the loss the FA inhibition on NOB caused by excess aeration. At low temperature(13.0–17.6°C), the level of nitrite pathway rapidly increased from 19.8% to 90%, suggesting that nitritation was successfully started up at low temperature by applying syntrophic association of the FA inhibition and real-time control, and then this high level of nitrite pathway was stably maintained for as long as 233 days. Mechanism analysis shows that the establishment of nitritation was primarily the result of predominant ammonia-oxidizing bacteria developed in the nitrifying bacteria population compared to NOB. This was mainly due to a gradual reduction of nitrite amount that is available to provide energy for the growth of NOB,eventually leading to the elimination of NOB from the bacterial clusters in sequencing batch reactor sludge system.     

基于间歇饥饿的SNAD工艺运行

在室温下(22℃±3℃)用SBR反应器运行SNAD工艺,通过定期延长系统水力停留时间,营造间歇饥饿环境,探讨间歇饥饿策略下SNAD工艺的运行情况.结果表明,系统经过间歇饥饿运行后,好氧阶段末的NO_3~--N浓度降至8. 72 mg·L~(-1),亚硝酸盐积累率达到83. 18%,表明NOB活性得到了有效抑制,实现了亚硝化性能的提高;系统经过间歇饥饿运行后,好氧阶段末的亚氮与氨氮基质的比例得到调整,为后续厌氧氨氧化过程提供了合适底物,使出水氨氮浓度降至1. 0 mg·L~(-1)以下,同时由于出水硝氮浓度降低,总氮去除率达到了92. 07%左右,系统处理性能提高;通过测定功能菌活性,发现饥饿后亚硝化性能提高的主要原因是饥饿期AOB活性衰减速率低于NOB及恢复期前期AOB活性恢复速率显著高于NOB.     

不同曝气量对SBBR短程硝化微生物特性及氮转化的影响

在实验室规模的序批式生物膜反应器(SBBR)中研究了不同曝气量(7.2、12.0、15.6L·h-1,对应反应器中平均溶解氧浓度分别为0.5、0.8、1.2mg·L-1)下生物膜的生物特性变化及短程硝化过程规律.结果表明:减小曝气量使反应器内溶解氧浓度降低,将导致生物膜的总生物量下降,生物膜中氨氧化菌逐渐成为优势菌,无论数量还是生物活性均高于亚硝酸氧化菌,利于亚硝酸盐积累;在一个反应周期中,生物膜对溶解氧需求的分配是不同的,曝气初期溶解氧主要用于异氧菌对COD的降解,其后用于氨氮转化.根据上述规律,提出在短程硝化过程中采用"梯级递减式曝气"供氧新策略,即在反应初期保持一种较大的曝气量,提高反应器溶解氧浓度,促进COD快速降解,随后保持一种小曝气量使反应器中溶解氧维持较低的浓度,从而促进亚硝酸盐积累及优化供氧效率.     

实际垃圾渗滤液短程生物脱氮的常温实现及低温维持

采用单级UASB-SBR生化系统处理实际垃圾渗滤液,主要考察了常、低温条件下,该生化系统短程生物脱氮的长期稳定性,同时研究了SBR内短程硝化的实现机理及微生物种群特性.598d试验结果表明:单级UASB-SBR生化系统对渗滤液内COD,NH4+-N和TN的去除率分别为92.0%,99.2%和98.0%以上,实现渗滤液内有机物和氮的深度去除.经过116d运行,SBR系统实现了短程硝化,亚硝积累率(NAR)达到90%以上,此后稳定运行,成功跨越2个冬季,15℃以下共计171d,最低温度为10.2℃.游离氨(FA)和过程控制的协同作用是实现与维持SBR 内短程硝化的决定因素.荧光原位杂交(FISH)技术检测表明:氨氧化菌(AOB)已经成为SBR硝化菌群中的优势菌属.扫描电子显微镜(SEM)检测表明:AOB菌属以活椭球状亚硝化球菌属(Nitrosococcus)和杆状亚硝化单胞菌属(Nitrosomonas)为主.     

污水处理系统中硝化菌的菌群结构和动态变化

研究分析了4种不同工艺类型的城市污水处理厂中氨氧化细菌(AOB)和亚硝酸盐氧化细菌(NOB)的丰度及菌群结构.实时定量PCR结果表明4种工艺中AOB菌群的丰度范围为8.56×106~4.46×107cells/gMLSS;NOB菌群的丰度为3.37×108~1.53×109cells/gMLSS.每个工艺中Nitrospira都是优势NOB,占NOB菌群的88% 以上. A2O工艺冬季AOB和Nitrospira丰度比夏季均有所降低,这是导致冬季生物脱氮效果变差的主要原因.基于 amoA基因的系统发育分析结果显示所有的序列属于Nitrosomonas,其中Nitrosomonas oligotropha cluster 占克隆文库的60.1%,是AOB 种群中的优势菌属,Nitrosomonas-like cluster和 Nitrosomonas europaea cluster次之,分别占克隆文库的29.6%和9.1%.N. europaea cluster只在A2O工艺中出现,且在A2O工艺夏季污泥样品克隆文库中达到44.7%.低DO运行使N. europaea cluster成为优势AOB是A2O工艺夏季出现较高亚硝酸盐积累率的主要原因.研究结果证实了城市污水处理厂中优势AOB和NOB分别为Nitrosomonas和Nitrospira,硝化菌群占总菌群的1%~7%,其丰度、相对含量和菌群结构是影响硝化效果的主要因素.