II 摘要 厌氧氨氧化(anammox)技术具有脱氮负荷高和运行成本低的优势，已成功应用 于高氨氮废水处理。重金属镍和无机盐在诸多高氨氮废水（如城市生活污水，工 业废水，垃圾渗滤液等）中共存，anammox 应用于该类废水脱氮处理时面临可能 的抑制问题。然而，很少报道 Ni(II)和无机盐对 anammox 工艺性能和微生物多样 性的复合效应。本学位报告即针对此开展了相关研究，以期为其实际应用提供参 考。主要研究内容和结论为： 探究了 Ni(II)对 anammox –升流式厌氧污泥床（UASB）系统的短期和长期脱 氮性能的影响。短期暴露试验结果表明，Ni(II) 的半抑制浓度(IC 50 )为 14.6 mg·L -1 ； 连续流试验（150 天）表明，经过缓慢增加 Ni(II) 浓度至 10 mg·Ni 2+ ·L -1 驯化, UASB 平均氮去除效率（NRE）达到 93%。在颗粒污泥特性方面，沉降速率、 比厌氧氨 氧化活性（SAA）和胞外聚合物（EPS）的含量随着镍浓度的升高呈降低之趋势； 而细胞色素 c 的含量则持续升高。在 0.3 mg·Ni 2+ ·L -1 短期暴露时即出现了活性抑制 效应；但是长期连续流运行和驯化 150 天后，anammox 体系可以耐受高达 10 mg·Ni 2+ ·L -1 的镍胁迫。16S rRNA 高通量测序结果显示 Ni(II)对于 anammox 反应 器中微生物种群的多样性有负面作用。在 0-10 mg·L -1 所试 Ni(II) 浓度范围内， Candidatus Kuenenia 的相对丰度从 36.23%下降至 28.46%。 研究了Ni(II) 和无机盐对 anammox-UASB 连续流运行条件下的脱氮能力的影 响及其受抑制后的恢复效果（包括工艺性能、颗粒特性、微生物种群变化）。结果 表明，在 0.2 mg·Ni 2+ ·L -1 和 20 g·NaCl·L -1 联合作用下，系统的脱氮性能严重失稳， 但在运行 170 天后系统性能逐渐恢复， NRE 和 NRR 分别达到 77.1%和 1.18 kg·N杭州师范大学硕士学位报告 摘要 III m 3 ·d -1 。三维荧光和红外光谱分析表明，无机盐和 Ni(II) 改变了 anammox 颗粒污 泥胞外聚合物（EPS）的分泌量和成分。用 Stover–Kincannon 模型、一阶模型和 二阶模型等数学模型模拟了反应器在不同时期的脱氮行为，取得了较好的拟合效 果。16S rRNA 高通量测序技术发现 Ca._Kuenenia 为所有试验阶段的优势菌属；当 SAA 降低了 98%，Ca._Kuenenia 的相对丰度降低了 18.1%。 关键词：厌氧氨氧化；镍；盐度；颗粒特性；菌群结构杭州师范大学硕士学位报告 ABSTRACT IV ABSTRACT Anaerobic ammonium oxidation (anammox) technology has some advantages of high nitrogen removal rates, low operational costs, and small footprints, and has been successfully implied to treat high-strength ammonium wastewater. However, very little is known about the synthetic toxicity of anammox technogy performances and microorganism diversity from divalent nickel and salinity. 1、In the present study, we revealed the short- and long-term influences of divalent nickel on anammox in an upflow anaerobic sludge blanket (UASB) reactor. The results showed that the half maximal inhibitory concentration (IC 50 ) of Ni(II) on anammox biomass was 14.64 mg L -1 in short-term exposure. The continuous-flow experiment was observed during the 150 days, and illustrated that after domestication, nitrogen removal efficiency was up to 93±0.03% under the addition of 10 mg·Ni 2+ ·L -1 . The settling velocity, SAA and content of EPS were decreased as the increase of the concentration of Ni(II) roughly, nevertheless, the content of heme c increased when the concentration of Ni(II) raised. It was implied that short-term exposure to Ni(II) had an adverse impact on anammox biomass, yet the anammox system could tolerate Ni(II) stress to 10 mg·L -1 after acclimation during the continuous-flow operation for 150 days. High-throughput sequencing results indicated that the presence of Ni(II) had an impact on the microbial community composition in the anammox reactor, especially Candidatus Kuenenia. At Ni(II) concentrations of 0-10 mg·L -1 , the relative abundance of Candidatus Kuenenia decreased from 36.23% to 28.46%. 2、Effects of the continuous addition of a high-concentration of salinity and Ni(II) on the characteristics of anammox as well as on the variety and recovery of their ability杭州师范大学硕士学位报告 ABSTRACT V removing nitrogen in an upflow anaerobic sludge blanket reactor (UASB). This is the first study to examine the changes in quantity and composition of the extracellular polymeric substances (EPS) in an anammox-UASB system with different treatment.The operation performance and the sludge characteristics was also investigated. The results showed that after a 170-day acclimation to the addition of 0.2 mg·Ni 2+ ·L -1 and 20 g ·NaCl·L -1 , the ability to remove nitrogen was mainly recovered, and NRE and NRR were reached to 77.08% and 1.18 kg·N·m 3 ·d -1 . Through 3D-EEM and FT-IR spectroscopy analysis, the introduced salinity and Ni(II) made a great change in the quantity and composition of the EPS in anammox granule. We used the mathematic model to simulate the behavior of anammox system in different phases, and investigated 16S rRNA gene of bacteria using the high throughput sequencing technology in a laboratory-scale up-flow anaerobic sludge blanket (UASB) bioreactor. Stover– Kincannon model, the First order model, the Second order model used to simulate the seven stages. The outcomes were revealed, (1) Planctomycetes Ca.Kuenenia is the dominant microorganism in all samples; (2) the abundance of Ca.Kuenenia only decreased by 18.08% as anammox activity reduced by 98%. Keywords：Anammox; Nickel;Salinity;Granul properties; Microbial diversity杭州师范大学硕士学位报告 目录 VI 目录 致谢...................................................................................................................................I 摘要..................................................................................................................................II ABSTRACT ................................................................................................................... IV 目录................................................................................................................................VI 1 引论 ............................................................................................................................... 1 1.1 含镍含盐含氨废水 .................................................................................................... 1 1.1.1 含镍含盐含氨废水的来源和特征 ................................................................. 1 1.1.2 含镍含盐含氨废水处理研究现状 ................................................................. 2 1.2 厌氧氨氧化（anammox）菌 .................................................................................... 3 1.2.1 Anammox 菌种 ............................................................................................... 3 1.2.2 Anammox 菌与生长基质 ............................................................................... 4 1.2.3 Anammox 菌与基质抑制 ............................................................................... 5 1.2.4 Anammox 菌与温度 ....................................................................................... 5 1.2.5 Anammox 菌与 pH 值..................................................................................... 7 1.2.6 Anammox 菌与溶解氧浓度及光照 ............................................................... 7 1.2.7 Anammox 菌与生长速率 ............................................................................... 8 1.3 Anammox 工艺应用现状 .......................................................................................... 8 1.4 UASB 反应器 ............................................................................................................ 8 1.5 研究目的和意义、内容、目标及方法 ...................................................................11 1.5.1 研究目的和意义 ............................................................................................11 1.5.2 研究内容 .......................................................................................................11 1.5.3 研究目标 ...................................................................................................... 12 1.5.4 研究方法 ...................................................................................................... 12 2. Ni(II)对 anammox-UASB 系统脱氮性能和微生物种群的影响 ............................. 14 2.1 概述 .......................................