回流比对厌氧氨氧化系统的影响及作用机制研究

doi:10.13560/j.cnki.biotech.bull.1985.2025-0719

摘要/Abstract

摘要：

目的探究回流比对厌氧氨氧化系统脱氮性能的影响，并揭示其作用机制。方法设定三组回流比（0%、50%和100%），测定不同回流比时厌氧氨氧化系统脱氮性能、颗粒污泥特性、微生物群落结构和氮代谢相关功能基因。结果当回流比为100%时，系统稳定阶段的平均总氮去除率和平均NRR分别提高了9.32%和36.09 mg/（L‧d），脱氮性能更稳定，∆NO₂^--N/∆NH₄⁺-N和∆NO₃^--N/∆NH₄⁺-N更接近厌氧氨氧化理论值。出水氧化还原电位更低，改善了系统中的氧环境。颗粒污泥颜色更红，颗粒化程度提高，上浮现象明显减少。颗粒污泥中紧密结合型多糖和松散结合型多糖含量分别增加了22.59和24.00 mg/g VSS，形成了更稳定的抗剪切结构。颗粒污泥表面的C-O和C=C官能团增加，加快了系统中的电子转移。高通量测序表明，Candidatus Brocadia维持较高的相对丰度（38.61%），Denitratisoma的相对丰度增加了4.13%，同时，氮代谢相关功能基因narG、napA、narZ和nirS出现了上调。结论适当的回流比可有效提高厌氧氨氧化系统脱氮性能和运行稳定性。

关键词: 回流比, 厌氧氨氧化, 颗粒污泥, 菌群结构, 功能基因, 作用机制

Abstract:

Objective To explore the effect of reflux ratio on denitrification performance of anaerobic ammonia oxidation system and reveal its mechanism of action. Method Three reflux ratios (0%, 50%, and 100%) were set to determine the denitrification performance, granular sludge characteristics, microbial community structure, and nitrogen metabolism related functional genes of the anaerobic ammonia oxidation system at different reflux ratios. Result When the reflux ratio was 100%, the average total nitrogen removal efficiency and the average NRR (nitrogen removal rate) during the stable phase of the system increased by 9.32% and 36.09 mg/(L‧d), respectively, with more stable nitrogen removal performance. The ratios of ∆NO₂^--N/∆NH₄⁺-N and ∆NO₃^--N/∆NH₄⁺-N were closer to the theoretical values of anammox. The effluent had a lower oxidation-reduction potential, which improved the oxygen environment in the system. The color of granular sludge was redder, the degree of granulation increased, and the phenomenon of upward floating significantly reduced. The contents of tightly bound polysaccharides and loosely bound polysaccharides within the granules increased by 22.59 and 24.00 mg/g VSS, respectively, forming a more stable shear-resistant structure. The increase of C-O and C=C functional groups on the surface of granular sludge accelerated the electron transfer in the system. High-throughput sequencing showed that Candidatus Brocadia maintained a high relative abundance (38.61%), while the relative abundance of Denitratisoma increased by 4.13%. Meanwhile,nitrogen metabolism-related functional genes narG, napA, narZ and nirS were upregulated Conclusion An appropriate reflux ratio can effectively improve the denitrification performance and operational stability of anaerobic ammonia oxidation systems.

Key words: reflux ratio, anammox, granule sludge, microbial community structure, functional gene, mechanism of action

陈仔龙, 张超, 郝宛婷, 宋贤威, 陈欣怡, 卢璐, 李鹏飞, 朱易春. 回流比对厌氧氨氧化系统的影响及作用机制研究[J]. 生物技术通报, 2026, 42(2): 102-112.

CHEN Zi-long, ZHANG Chao, HAO Wan-ting, SONG Xian-wei, CHEN Xin-yi, LU Lu, LI Peng-fei, ZHU Yi-chun. Study on the Influence and Mechanism of Reflux Ratio on Anaerobic Ammonia Oxidation System[J]. Biotechnology Bulletin, 2026, 42(2): 102-112.

图/表 9

图1 UASB装置示意图

Fig. 1 Schematic diagram of UASB device

图2 不同回流比下氮的转化和去除A：氨氮的变化；B：亚硝态氮的变化；C：硝态氮和总氮的变化；D：NLR和NRR的变化

Fig. 2 Nitrogen transformation and removal under different reflux ratiosA: Changes in ammonia nitrogen; B: changes in nitrite nitrogen; C: changes in nitrate nitrogen and total nitrogen; D: changes in NLR and NRR

图3 不同回流比下化学计量比和氧化还原电位的变化A：化学计量比的变化；B：氧化还原电位的变化

Fig. 3 Variations in stoichiometric ratios and oxidation-reduction potential under different reflux ratiosA: Changes in stoichiometric ratio; B: changes in redox potential

图5 不同回流比下污泥EPS组分及含量的变化R1、R2、R3分别代表回流比为0%、50%、100%，下同

Fig. 5 Variations in EPS composition and content of granule sludge under different reflux ratiosR1, R2, R3 indicate the reflux ratio of 0%, 50%, and 100%, respectively. The same below

图6 不同回流比下污泥表面官能团的变化

Fig. 6 Variations in surface functional groups of granule sludge under different reflux ratios

表1 不同回流比下Alpha多样性指数的变化

Table 1 Variations in alpha diversity indices under different reflux ratios

	OTUs	Shannon	Chao1	Ace	Simpson	Shannoneven
R1	852	3.328	866	887	0.163	0.493
R2	836	3.447	850	869	0.145	0.512
R3	808	3.273	828	854	0.158	0.489

图7 不同回流比下微生物群落结构的变化A：门水平的相对丰度；B：属水平的相对丰度

Fig. 7 Variations in microbial community structure under different reflux ratiosA: Relative abundance at the phylum level; B: relative abundance at the genus level

图8 氮代谢相关功能基因的变化

Fig. 8 Variations in nitrogen metabolism-related functional gene

图9 出水回流对厌氧氨氧化系统的作用机制

Fig. 9 Underlying mechanism of effluent reflux in anammox systems

参考文献 31

[1]	Fu JR, Liao H, Wang ZL, et al. Effect of simultaneously addition of Fe³⁺ and reed straw biochar on nitrogen removal efficiency and GWP for Anammox [J]. Chem Eng J, 2025, 506: 160090.
[2]	雷雪艳, 朱易春, 张超, 等. 零价铁耦合厌氧氨氧化系统高效同步脱氮除磷 [J]. 化工进展, 2025, 44(6): 3132-3143.
	Lei XY, Zhu YC, Zhang C, et al. Efficient synchronous nitrogen and phosphorus removal in zero valent iron coupled anaerobic ammonia oxidation system [J]. Chem Ind Eng Prog, 2025, 44(6): 3132-3143.
[3]	Chen XY, Liu LJ, Bi YM, et al. A review of anammox metabolic response to environmental factors: Characteristics and mechanisms [J]. Environ Res, 2023, 223: 115464.
[4]	Paritosh K, Kesharwani N. Biochar mediated high-rate anaerobic bioreactors: a critical review on high-strength wastewater treatment and management [J]. J Environ Manag, 2024, 355: 120348.
[5]	Cema G, Gutwiński P, Ziembińska-Buczyńska A, et al. Evaluation of the long-term effect of Cr(Ⅲ), Zn(Ⅱ), Cd(Ⅱ), Cu(Ⅱ), Ni(Ⅱ) and Pb(Ⅱ) on the anammox process in a continuous-flow anaerobic membrane-assisted bioreactor (AnMBR) [J]. Water Air Soil Pollut, 2025, 236(2): 86.
[6]	Yang JH, Huang DQ, Wu GG, et al. Quorum sensing-mediated microecological homeostasis in anammox consortia [J]. J Hazard Mater, 2025, 492: 138285.
[7]	傅慧敏, 冷济轩, 翁勋, 等. 出水回流对Anammox颗粒特性及其微生物群落的影响 [J]. 中国环境科学, 2022, 42(4): 1663-1671.
	Fu HM, Leng JX, Weng X, et al. The effects of effluent reflux on characteristics and microbial community of anammox granular sludge [J]. China Environ Sci, 2022, 42(4): 1663-1671.
[8]	Zhang ZX, Xing W, Lu J, et al. Nitrogen removal and nitrous oxide emission in the partial nitritation/anammox process at different reflux ratios [J]. Sci Total Environ, 2024, 906: 167520.
[9]	陆明羽, 殷记强, 姚凤根, 等. 回流比对DN-PN-ANAMMOX工艺处理中晚期垃圾渗滤液的影响 [J]. 环境科学研究, 2021, 34(4): 996-1005.
	Lu MY, Yin JQ, Yao FG, et al. Effect of recycle ratios on partial nitrification-anammox with pre-denitrification of mature leachate treatment [J]. Res Environ Sci, 2021, 34(4): 996-1005.
[10]	Yan ZC, Zeng R, Yang H. Sustainable treatment of high-ammonia-nitrogen organic wastewater via anaerobic ammonium oxidation (anammox) combined with effluent recirculation/micro-aeration [J]. Sustainability, 2025, 17(13): 5926.
[11]	Qi PQ, Li J, Dong HY, et al. Performance of anammox process treating nitrogen-rich saline wastewater: Kinetics and nitrite inhibition [J]. J Clean Prod, 2018, 199: 493-502.
[12]	Du R, Cao SB, Zhang HY, et al. Flexible nitrite supply alternative for mainstream anammox: advances in enhancing process stability [J]. Environ Sci Technol, 2020, 54(10): 6353-6364.
[13]	雷雪艳, 晏雯雯, 张超, 等. 湿干比对不同人工快速渗滤系统处理厌氧折流板反应器出水的影响 [J]. 江西冶金, 2023, 43(3): 259-264.
	Lei XY, Yan WW, Zhang C, et al. Effect of wet/dry ratio on anaerobic baffled reactor effluent treatment with different constructed rapid infiltration systems [J]. Jiangxi Metall, 2023, 43(3): 259-264.
[14]	Xu DD, Fan JH, Pan C, et al. Dimension effect of anammox granule: Potential vs performance [J]. Sci Total Environ, 2021, 795: 148681.
[15]	Tang X, Guo YZ, Jiang B, et al. Metagenomic approaches to understanding bacterial communication during the anammox reactor start-up [J]. Water Res, 2018, 136: 95-103.
[16]	Chang BZ, Huang XL, Chen DZ, et al. How biofilm and granular sludge cope with dissolved oxygen exposure in anammox process: Performance, bioaccumulation characteristics and bacterial evolution [J]. J Environ Manag, 2025, 373: 123986.
[17]	Feng CJ, Lotti T, Lin YM, et al. Extracellular polymeric substances extraction and recovery from anammox granules: Evaluation of methods and protocol development [J]. Chem Eng J, 2019, 374: 112-122.
[18]	高雪健, 张杰, 李冬, 等. 不同基质浓度对ANAMMOX菌短期储存的影响 [J]. 环境科学, 2018, 39(12): 5587-5595.
	Gao XJ, Zhang J, Li D, et al. Effects of different substrate concentrations on the short-term storage of ANAMMOX bacteria [J]. Environ Sci, 2018, 39(12): 5587-5595.
[19]	Wang S, Huang XX, Liu LJ, et al. Understanding the mechanism in aggregation ability between aerobic and anammox granular sludge from the perspective of exopolysaccharides [J]. J Water Process Eng, 2020, 38: 101629.
[20]	Zhang LH, Zhang J, Hu X. Analyzing the nitrogen removal performance and cold adaptation mechanism of immobilized cold-acclimation ANAMMOX granules at low temperatures [J]. Water Environ Res, 2024, 96(2): e10985.
[21]	Zhen JY, Zheng M, Wei W, et al. Extracellular electron transfer (EET) enhanced anammox process for progressive nitrogen removal: a review [J]. Chem Eng J, 2024, 482: 148886.
[22]	Witkabel P, Abendroth C. A systematic literature review of microbial anammox consortia in UASB/EGSB-reactors [J]. Chemosphere, 2024, 367: 143630.
[23]	Wang JX, Liang JD, Sun L, et al. Achieving reliable partial nitrification and anammox process using polyvinyl alcohol gel beads to treat low-strength ammonia wastewater [J]. Bioresour Technol, 2021, 324: 124669.
[24]	Lawson CE, Wu S, Bhattacharjee AS, et al. Metabolic network analysis reveals microbial community interactions in anammox granules [J]. Nat Commun, 2017, 8: 15416.
[25]	Pushpakumara BLDU, Tandon K, Willis A, et al. Unravelling microalgal-bacterial interactions in aquatic ecosystems through 16S rRNA gene-based co-occurrence networks [J]. Sci Rep, 2023, 13: 2743.
[26]	Choi D, Yun W, Choi Y, et al. Insights into thiosulfate-driven partial denitrification synergistically mediated by anaerobic ammonium oxidation: Biosynthesized signaling molecules and enzymatic collaboration [J]. Chem Eng J, 2025, 506: 160069.
[27]	Li JP, Zuo XT, Chen QQ, et al. Genome-resolved metagenomic analysis reveals a novel denitrifier with truncated nitrite reduction pathway from the genus SC-I-84 [J]. Water Res, 2025, 282: 123598.
[28]	雷雪艳, 朱易春, 张超, 等. 常温有氧低基质下厌氧氨氧化二次启动及脱氮效能 [J]. 中国给水排水, 2025, 41(1): 16-24.
	Lei XY, Zhu YC, Zhang C, et al. Secondary start-up and nitrogen removal performance of anammox under room temperature, aerobic, and low concentration substrate conditions [J]. China Water Wastewater, 2025, 41(1): 16-24.
[29]	Zhen JY, Wang ZB, Ni BJ, et al. Synergistic integration of anammox and endogenous denitrification processes for the simultaneous carbon, nitrogen, and phosphorus removal [J]. Environ Sci Technol, 2024, 58(24): 10632-10643.
[30]	Lin BC, Hu TQ, Xu ZH, et al. Stratified biofilm structure of MABR enabling efficient ammonia removal from aquaculture medicated bath wastewater [J]. Water Res, 2025, 277: 123326.
[31]	Xiao Y, Yin XJ, Chen LJ, et al. Effects of illumination on nirS denitrifying and anammox bacteria in the rhizosphere of submerged macrophytes [J]. Sci Total Environ, 2021, 760: 143420.