基于代谢组的厌氧氨氧化菌群对温度的响应机制

【背景】在环境工程领域中,大多数耐冷菌从活性污泥中分离得到。了解活性污泥菌群对低温的响应有助于耐冷菌的驯化培养。【目的】以厌氧氨氧化污泥菌群作为研究对象,研究温度对厌氧氨氧化菌群代谢通路与代谢产物的影响,以期初步阐释厌氧氨氧化菌群低温响应机理。【方法】在25°C与35°C条件下驯化培养厌氧氨氧化污泥,研究温度对反应器脱氮效能、菌群活性与生长以及群落结构的影响,通过代谢组学比较两个温度下厌氧氨氧化菌群代谢物丰度以及代谢通路活性。【结果】虽然低温导致厌氧氨氧化菌群CO_2固定、TCA循环与丙酮酸代谢的下调,进而导致氮去除活性以及增长速率显著下降,但菌群RNA合成水平、腐胺与信号分子合成上调,从而通过转录调控、调控膜脂组成与改变膜结构的方式,调控菌群代谢以适应低温环境。【结论】从分子机理的角度探究了厌氧氨氧化污泥菌群适应低温环境的生理机制,首次阐释了腐胺与信号分子在污泥菌群适应低温过程中的重要作用。     

海洋浮霉状菌多样性与生态学功能研究进展

浮霉状菌为一个独立的细菌门,具独特的细胞结构与基因组组成,是一类重要的环境微生物。有趣的是,浮霉状菌在海洋环境中具多样的生态学功能,如参与海洋碳循环、海洋厌氧氨氧化过程、矿物质富聚作用、及藻赤潮等,现已成为国际海洋环境微生物研究最热门的领域之一。然而,目前国内对该细菌门的研究进展尚无综合报道,由此,本文综合最新和最重要的文献综述了海洋浮霉状菌多样性及生态学功能的最新进展,并结合自己的工作对该领域未来的研究进行了展望。     

烟气脱硝尾液厌氧氨氧化处理中微生物群落结构分析

采用厌氧序批式反应器处理火电厂烟气脱硝尾液并研究厌氧氨氧化微生物群落结构。通过扫描电镜观察发现,优势菌群为椭圆形的球菌且具有下凹的火山口状,直径约为0.7μm。通过污泥总DNA提取、PCR扩增、阳性克隆验证和测序等分子生物学手段得到反应器内菌群16S r RNA基因序列,建立系统发育树和克隆文库。结果表明,在细菌通用引物27F-1 492R PCR扩增的体系中,得到85个克隆子,分为21个OTU,各菌群所占的比例为:变形菌门Proteobacteria:61.18%;酸杆菌门Acidobacteria:17.65%;绿菌门Chlorobi:8.24%;绿弯菌门Chlorofexi:5.88%;芽单胞菌门Gemmatimonadetes:3.53%;硝化螺旋菌门Nitrospirae:2.35%;浮酶状菌门Planctomycetes:1.18%。而在厌氧氨氧化菌特异引物pla46rc-630r;AMX368-AMX820 PCR扩增的体系中有45个克隆子,分为3个OTU,其中Candidatus brocadia sp.占有95.6%,未知菌种4.4%。     

有机碳源下废水厌氧氨氧化同步脱氮除碳

为明确有机碳源胁迫下,厌氧氨氧化反应器的同步脱氮除碳规律及功能微生物群落结构的动态变化,采用成功启动的厌氧氨氧化UASB反应器,通过逐步提升进水有机负荷,探究有机碳源下废水厌氧氨氧化同步脱氮除碳。研究表明,当进水化学需氧量(Chemical oxygen demand,COD)浓度从172 mg/L升至620 mg/L,反应器维持较高的脱氮效率,氨氮和总氮去除率均在85%以上,并对COD具有平均56.6%的去除率,高浓度COD未对Anammox菌活性构成显著抑制作用。聚合酶链式反应和变性梯度凝胶电泳(PCR-DGGE)图谱和割胶测序结果表明,变形菌门Proteobacteria、浮霉菌门Planctomycetes、绿曲挠菌门Chloroflexi以及绿菌门Chlorobi等微生物共存于同一反应体系中,推测反应器内存在复杂的脱氮除碳途径。而且,代表厌氧氨氧化的部分浮霉菌门微生物能耐受高浓度有机碳源,在高有机负荷下依旧发挥着高效的脱氮作用,为反应器高效脱氮提供了保障。     

Anammox反应器启动过程中颗粒污泥性状变化特性

以厌氧颗粒污泥作为接种物,通过185 d的运行,成功启动了上流式厌氧氨氧化污泥床(Upflow anaerobic sludge blanket,UASB)反应器。反应器的进水氨氮与亚硝氮浓度分别提升至224 mg/L和255 mg/L,容积氮去除速率提升至3.76 kg/(m3·d)。采用红外光谱、扫描电镜和透射电镜等对厌氧氨氧化颗粒污泥的性状进行观察,发现颗粒污泥在启动过程中经历了污泥颗粒裂解到污泥颗粒重组的过程,且厌氧氨氧化颗粒污泥表面含有丰富的官能团,说明厌氧氨氧化颗粒污泥可能具有良好的吸附性能。采用宏基因组测序的方法对启动前后颗粒污泥的生态结构进行分析,发现原接种污泥优势菌群(变形菌门、厚壁菌门、拟杆菌门)丰度大幅减少,厌氧氨氧化菌所属的浮霉状菌门丰度则由1.59%提升到23.24%。     

Ladderane lipid distribution in four genera of anammox bacteria

Intact ladderane phospholipids and core lipids were studied in four species of anaerobic ammonium oxidizing (anammox) bacteria, each representing one of the four known genera. Each species of anammox bacteria contained C(18) and C(20) ladderane fatty acids with either 3 or 5 linearly condensed cyclobutane rings and a ladderane monoether containing a C(20) alkyl moiety with 3 cyclobutane rings. The presence of ladderane lipids in all four anammox species is consistent with their putative physiological role to provide a dense membrane around the anammoxosome, the postulated site of anammox catabolism. In contrast to the core lipids, large variations were observed in the distribution of ladderane phospholipids, i.e. different combinations of hydrophobic tail (ladderane, straight chain and methyl branched fatty acid) types attached to the glycerol backbone sn-1 position, in combination with different types of polar headgroup (phosphocholine, phosphoethanolamine or phosphoglycerol) attached to the sn-3 position. Intact ladderane lipids made up a high percentage of the lipid content in the cells of "Candidatus Kuenenia stuttgartiensis", suggesting that ladderane lipids are also present in membranes other than the anammoxosome. Finally, all four investigated species contained a C(27) hopanoid ketone and bacteriohopanetetrol, which, indicates that hopanoids are anaerobically synthesised by anammox bacteria.     

氮形态转化途径研究的新进展—厌气铵氧化及其应用前景

20世纪90年代初在污泥处理系统中发现了氮素形态转化的新途径－厌气铵氧化过程,厌气铵氧化过程是铵以亚硝酸根为电子受体在自养细菌参数下氧化成氮气的过程,但目前尚无土壤,河,湖,海底泥等自然系统中是否存在厌氧铵氧化过程的报道,由于该过程无需外加有机碳,耗氧和处理产生污泥少,用于污泥脱氮成本较低,具有很大潜力。     

Development of a simultaneous partial nitrification and anaerobic ammonia oxidation process in a single reactor

Up-flow oxygen-controlled biofilm reactors equipped with a non-woven fabric support were used as a single reactor system for autotrophic nitrogen removal based on a combined partial nitrification and anaerobic ammonium oxidation (anammox) reaction. The up-flow biofilm reactors were initiated as either a partial nitrifying reactor or an anammox reactor, respectively, and simultaneous partial nitrification and anammox was established by careful control of the aeration rate. The combined partial nitrification and anammox reaction was successfully developed in both biofilm reactors without additional biomass inoculation. The reactor initiated as the anammox reactor gave a slightly higher and more stable mean nitrogen removal rate of 0.35 (± 0.19) kg-N m⁻³ d⁻¹ than the reactor initiated as the partial nitrifying reactor (0.23 (± 0.16) kg-N m⁻³ d⁻¹). FISH analysis revealed that the biofilm in the reactor started as the anammox reactor were composed of anammox bacteria located in inner anoxic layers that were surrounded by surface aerobic AOB layers, whereas AOB and anammox bacteria were mixed without a distinguishable niche in the biofilm in the reactor started as the partial nitrifying reactor. However, it was difficult to efficiently maintain the stable partial nitrification owing to inefficient aeration in the reactor, which is a key to development of the combined partial nitrification and anammox reaction in a single biofilm reactor.     

Partial nitritation and anammox of a livestock manure digester liquor and analysis of its microbial community

A swim-bed reactor for partial nitritation with polymeric coagulant treatment and an UASB reactor for anammox were applied to the treatment of livestock manure digester liquor. The partial nitritation was maintained for 32 days under a 1.6 kg N/m³/d nitrogen loading rate (NLR) with an average conversion efficiency of 51%, and achieved 1.65 kg N/m³/d of the maximum nitrite production rate under 2.58 kg N/m³/d of NLR. Although 200 mg/L of TOC remained in the effluent of the partial nitritation reactor, the anammox nitrogen removal rate was not significantly decreased and a relatively high rate of 2.0 kg N/m³/d was obtained under a NLR of 2.2 kg N/m³/d. 16S rRNA gene analysis showed that Nitrosomonas and KSU-1 were dominant in the partial nitritation and anammox reactor, respectively. The results of this study demonstrated that the partial nitritation-anammox process has possibility of applying to the nitrogen removal of livestock manure digester liquor.     

厌氧氨氧化菌的代谢途径及其关键酶的研究进展

作为新型生物脱氮工艺的代表,厌氧氨氧化反应的代谢功能菌——厌氧氨氧化(Anammox)菌也逐渐成为研究的热点。本文介绍了10种Anammox菌的发现过程,阐述了Anammox菌的关键酶的研究进展。通过对Candidatus"Brocadia anammoxidans"和Candidatus"Kuenenia stuttgartiensis"菌的代谢途径的分析和推理,综述了化学计量模型、生化反应模型和能量代谢模型,并提出了一种可能存在的基于关键酶的厌氧氨氧化菌微界面代谢新途径。