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Article(id=1234106389128344355, tenantId=1146029695717560320, journalId=1234093305789726721, issueId=1234106384963400440, articleNumber=null, orderNo=null, doi=null, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1730995200000, receivedDateStr=2024-11-08, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1772163491756, onlineDateStr=2026-02-27, pubDate=1750348800000, pubDateStr=2025-06-20, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772163491756, onlineIssueDateStr=2026-02-27, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772163491756, creator=13701087609, updateTime=1772163491756, updator=13701087609, issue=Issue{id=1234106384963400440, tenantId=1146029695717560320, journalId=1234093305789726721, year='2025', volume='45', issue='6', pageStart='2961', pageEnd='3552', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1772163490763, creator=13701087609, updateTime=1772163969484, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1234108392948682946, tenantId=1146029695717560320, journalId=1234093305789726721, issueId=1234106384963400440, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1234108392948682947, tenantId=1146029695717560320, journalId=1234093305789726721, issueId=1234106384963400440, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=3001, endPage=3009, ext={EN=ArticleExt(id=1234106389979788093, articleId=1234106389128344355, tenantId=1146029695717560320, journalId=1234093305789726721, language=EN, title=Highly efficient nitrogen and phosphorus removal by a novel denitrifying phosphorus removal system operated in an anaerobic-anoxic-aerobic-anoxic mode, columnId=1234106386360103680, journalTitle=China Environmental Science, columnName=Water Pollution Control, runingTitle=null, highlight=null, articleAbstract=

Cooperative strategies that mitigate competition among dominant functional microorganisms are crucial for efficient nitrogen and phosphorus removal in wastewater treatment. This study investigated a novel single-stage sequencing batch reactor operating under an anaerobic/anaerobic/oxic/anaerobic (A/A1/O/A2) mode for 100days to regulate the dynamic balance of phosphorus-accumulating organisms (PAOs), denitrifying PAOs (DPAOs), and denitrifying glycogen-accumulating organisms (DGAOs). The optimized system, featuring a recycle loop and reduced aerobic phase duration, achieved nitrogen and phosphorus removal efficiencies of (95.13%±0.35%) and (94.70%±0.96%), respectively. Mechanistic analysis suggested that the A1 phase created an anoxic environment conducive to DPAO-mediated denitrifying phosphorus removal, while the A2 phase supported DGAO-driven denitrifying nitrogen removal using polyhydroxyalkanoates (PHAs) and glycogen (Gly). Extracellular polymeric substance (EPS) analysis revealed increases of 35.38mg/gVSS in protein (PN) and 12.39mg/gVSS in polysaccharide (PS) content, enhancing sludge aggregation. Microbial community analysis demonstrated significant enrichment of Dechloromonas and Ca. Competibacter, with their abundances increasing from 2.24% and 1.53% in R1to 7.61% and 7.94% in R2, respectively. The A/A1/O/A2 mode effectively created a synergistic environment for key DPAOs and DGAOs, achieving superior nitrogen and phosphorus removal performance compared to conventional modes.

, correspAuthors=Dong LI, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Dong LI, Sai-yue QI, Si-bo FU, Yan-jun ZHU, Qin-yuan WANG, Jie ZHANG), CN=ArticleExt(id=1234106393079378851, articleId=1234106389128344355, tenantId=1146029695717560320, journalId=1234093305789726721, language=CN, title=厌氧/缺氧/好氧/缺氧模式DPR系统脱氮除磷性能, columnId=1234106386565624579, journalTitle=中国环境科学, columnName=水污染与控制, runingTitle=null, highlight=null, articleAbstract=

在废水的脱氮除磷过程中，通过合作避免优势功能微生物的共生竞争是深度脱氮除磷的有效策略.本研究采用单级序批式反应器以厌氧/缺氧/好氧/缺氧(A/A1/O/A2)的模式运行100d，调节聚磷菌(PAOs)、反硝化聚磷菌(DPAOs)和反硝化聚糖菌(DGAOs)的动态平衡，以期实现高效脱氮除磷.结果表明，采用A/A1/O/A2模式的新型反硝化除磷(DPR)系统为不同功能微生物创造了协同环境，通过增设回流装置和缩短有氧段时间的优化，氮去除效率(NRE)和磷去除效率(PRE)最终分别达到了(95.13%±0.35%)和(94.70%±0.96%)，实现了良好的脱氮除磷效果.机理分析表明，新型DPR系统中，好氧段前的A1段可能为DPAOs创造了良好的缺氧环境，DPAOs在此进行反硝化除磷，而在A2段，DGAOs可能在聚羟基烷链酸(PHAs)和糖原(Gly)驱动下实现内源反硝化脱氮.胞外聚合物(EPS)结果分析表明，相较于以A/O/A模式运行的R1，以A/A1/O/A2模式运行的R2中蛋白质(PN)和多糖(PS)含量分别提高了35.38和12.39mg/gVSS，污泥聚集性得到增强.此外，对微生物群落结构的分析表明，Dechloromonas和Ca.Competibacter的丰度分别从R1的2.24%、1.53%上升到R2的7.61%、7.94%和R3的4.62%、5.28%，A/A1/O/A2模式显著提高了主要DPAOs菌属和主要DGAOs菌属的丰度.

, correspAuthors=李冬, authorNote=null, correspAuthorsNote=
* 责任作者，教授，
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李冬(1976-)，女，辽宁丹东人，教授，博士，研究方向为水环境恢复理论及关键技术.发表论文200余篇..

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李冬(1976-)，女，辽宁丹东人，教授，博士，研究方向为水环境恢复理论及关键技术.发表论文200余篇..

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李冬(1976-)，女，辽宁丹东人，教授，博士，研究方向为水环境恢复理论及关键技术.发表论文200余篇..

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Operational conditions of the reactor

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反应器阶段阶段1(1~50d)阶段2(51~60d)阶段3(61~70d)阶段4(71~100d)
A(min)180150120120
R1A1(min)0404040
O(min)12012012060
A2(min)120808080
R2A1(min)40404040
O(min)12012012090
A2(min)80808080
R3A1(min)80404040
O(min)120120120120
A2(min)40808080
), ArticleFig(id=1234106402495591113, tenantId=1146029695717560320, journalId=1234093305789726721, articleId=1234106389128344355, language=CN, label=表1, caption=

反应器运行工况

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反应器阶段阶段1(1~50d)阶段2(51~60d)阶段3(61~70d)阶段4(71~100d)
A(min)180150120120
R1A1(min)0404040
O(min)12012012060
A2(min)120808080
R2A1(min)40404040
O(min)12012012090
A2(min)80808080
R3A1(min)80404040
O(min)120120120120
A2(min)40808080
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厌氧/缺氧/好氧/缺氧模式DPR系统脱氮除磷性能
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李冬 1, * , 齐赛月 1 , 傅思博 1 , 祝彦均 1 , 王沁源 1 , 张杰 1, 2
中国环境科学 | 水污染与控制 2025,45(6): 3001-3009
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中国环境科学 | 水污染与控制 2025, 45(6): 3001-3009
厌氧/缺氧/好氧/缺氧模式DPR系统脱氮除磷性能
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李冬1, * , 齐赛月1, 傅思博1, 祝彦均1, 王沁源1, 张杰1, 2
作者信息
  • 1.北京工业大学，水质科学与水环境恢复工程北京市重点实验室，北京 100124
  • 2.哈尔滨工业大学，城市水资源与水环境国家重点实验室，黑龙江 哈尔滨 150090

    • 李冬(1976-)，女，辽宁丹东人，教授，博士，研究方向为水环境恢复理论及关键技术.发表论文200余篇..

通讯作者:

* 责任作者，教授，
Highly efficient nitrogen and phosphorus removal by a novel denitrifying phosphorus removal system operated in an anaerobic-anoxic-aerobic-anoxic mode
Dong LI1, * , Sai-yue QI1, Si-bo FU1, Yan-jun ZHU1, Qin-yuan WANG1, Jie ZHANG1, 2
Affiliations
  • 1.Beijing Key Laboratory of Water Quality Science and Water Environment Restoration Project, Beijing University of Technology, Beijing 100124, China
  • 2.State Key Laboratory of Urban Water Resources and Water Environment, Harbin University of Technology, Harbin 150090, China
出版时间: 2025-06-20
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摘要
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在废水的脱氮除磷过程中，通过合作避免优势功能微生物的共生竞争是深度脱氮除磷的有效策略.本研究采用单级序批式反应器以厌氧/缺氧/好氧/缺氧(A/A1/O/A2)的模式运行100d，调节聚磷菌(PAOs)、反硝化聚磷菌(DPAOs)和反硝化聚糖菌(DGAOs)的动态平衡，以期实现高效脱氮除磷.结果表明，采用A/A1/O/A2模式的新型反硝化除磷(DPR)系统为不同功能微生物创造了协同环境，通过增设回流装置和缩短有氧段时间的优化，氮去除效率(NRE)和磷去除效率(PRE)最终分别达到了(95.13%±0.35%)和(94.70%±0.96%)，实现了良好的脱氮除磷效果.机理分析表明，新型DPR系统中，好氧段前的A1段可能为DPAOs创造了良好的缺氧环境，DPAOs在此进行反硝化除磷，而在A2段，DGAOs可能在聚羟基烷链酸(PHAs)和糖原(Gly)驱动下实现内源反硝化脱氮.胞外聚合物(EPS)结果分析表明，相较于以A/O/A模式运行的R1，以A/A1/O/A2模式运行的R2中蛋白质(PN)和多糖(PS)含量分别提高了35.38和12.39mg/gVSS，污泥聚集性得到增强.此外，对微生物群落结构的分析表明，Dechloromonas和Ca.Competibacter的丰度分别从R1的2.24%、1.53%上升到R2的7.61%、7.94%和R3的4.62%、5.28%，A/A1/O/A2模式显著提高了主要DPAOs菌属和主要DGAOs菌属的丰度.

脱氮除磷  /  反硝化除磷  /  内源反硝化  /  EPS

Cooperative strategies that mitigate competition among dominant functional microorganisms are crucial for efficient nitrogen and phosphorus removal in wastewater treatment. This study investigated a novel single-stage sequencing batch reactor operating under an anaerobic/anaerobic/oxic/anaerobic (A/A1/O/A2) mode for 100days to regulate the dynamic balance of phosphorus-accumulating organisms (PAOs), denitrifying PAOs (DPAOs), and denitrifying glycogen-accumulating organisms (DGAOs). The optimized system, featuring a recycle loop and reduced aerobic phase duration, achieved nitrogen and phosphorus removal efficiencies of (95.13%±0.35%) and (94.70%±0.96%), respectively. Mechanistic analysis suggested that the A1 phase created an anoxic environment conducive to DPAO-mediated denitrifying phosphorus removal, while the A2 phase supported DGAO-driven denitrifying nitrogen removal using polyhydroxyalkanoates (PHAs) and glycogen (Gly). Extracellular polymeric substance (EPS) analysis revealed increases of 35.38mg/gVSS in protein (PN) and 12.39mg/gVSS in polysaccharide (PS) content, enhancing sludge aggregation. Microbial community analysis demonstrated significant enrichment of Dechloromonas and Ca. Competibacter, with their abundances increasing from 2.24% and 1.53% in R1to 7.61% and 7.94% in R2, respectively. The A/A1/O/A2 mode effectively created a synergistic environment for key DPAOs and DGAOs, achieving superior nitrogen and phosphorus removal performance compared to conventional modes.

nitrogen and phosphorus removal  /  denitrifying phosphorus removal  /  endogenous denitrification  /  EPS
李冬, 齐赛月, 傅思博, 祝彦均, 王沁源, 张杰. 厌氧/缺氧/好氧/缺氧模式DPR系统脱氮除磷性能. 中国环境科学, 2025 , 45 (6) : 3001 -3009 .
Dong LI, Sai-yue QI, Si-bo FU, Yan-jun ZHU, Qin-yuan WANG, Jie ZHANG. Highly efficient nitrogen and phosphorus removal by a novel denitrifying phosphorus removal system operated in an anaerobic-anoxic-aerobic-anoxic mode[J]. China Environmental Science, 2025 , 45 (6) : 3001 -3009 .
正文
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作为人为控制废水中氮磷排放的关键一环，污水处理厂(WWTP)对市政污水中氮和磷的有效去除仍然面临缺乏碳源的挑战.低C/N比的情况与实现城市污水中氮磷的同步去除存在矛盾[1].因此反硝化除磷(DPR)工艺作为一种高效且低碳需求的生物技术，近年来受到了广泛的关注[2].
在传统DPR系统中，通常通过厌氧、缺氧和好氧3个阶段的交替作用来实现氮磷的同步去除.在厌氧阶段，主要功能菌反硝化聚磷菌(DPAOs)利用聚磷酸盐(poly-P)和糖原(Gly)分解产生的能量，从水中吸收挥发性脂肪酸，并将其转化为聚羟基烷链酸(PHAs)等胞内碳源，同时向废水中释放大量磷酸盐(PO43－-P)[3].在缺氧阶段，DPAOs以硝酸盐(NO3－-N)或亚硝酸盐(NO2－-N)代替氧气(O2)作为电子受体氧化PHAs并获得能量，从而从水中过量吸收PO43－-P，达到脱氮除磷的目的[4].然而，DPAOs使用的底物NO3－-N或NO2－-N不能直接从原废水中获得.它们需要由氨氧化菌(AOB)提供.AOB在原废水中转化氨氮(NH4+-N)，这需要一定的氧气.因此，在DPR系统中不可避免地存在好氧段以去除氮和磷.聚磷菌(PAOs)在厌氧条件下将外部碳源转化为PHAs进行储存，并在好氧段将氧气作为电子受体吸收过量的磷.由于DPR系统中氧气的存在，PAOs会大量增殖，这些PAOs不仅会增加能量消耗，还会与DPAOs争夺碳源造成竞争性抑制.
以往的研究经常在厌氧段和缺氧段之间添加一个好氧阶段(即A/O/A模式)[5].然而，这种先有氧再缺氧的方法往往会增强PAOs的优势.但如果将好氧段后移到较晚的阶段，会出现两个问题：第一，它不能为缺氧段的DPAOs提供底物，这可以通过建立再循环系统来解决.其次，未经进一步处理的好氧废水将不符合NO3－-N和NO2－-N的排放标准.作为脱氮除磷系统中重要的功能菌，反硝化聚糖菌(DGAOs)总是随着DPAOs的富集而增殖，并在系统中与其他脱氮除磷的功能菌相互影响.在厌氧和好氧/缺氧阶段，DGAOs利用与PAOs/DPAOs相似的代谢来促进内源反硝化，而不会积累聚磷酸盐[6].因此，如果在后移好氧段后，再增加一个缺氧段，为DGAOs提供合适环境，就可以进一步处理好氧废水中的NO3－-N和NO2－-N，同时还可以充分利用DPR系统中共存的碳源，最大化的平衡功能微生物.
本研究在后移好氧段后，再增加一个缺氧段，提出一种新的操作模式——厌氧-缺氧-好氧-缺氧(A/A1/O/A2)模式，旨在通过调节PAOs、DPAOs与DGAOs之间的相互作用，优化微生物群落的平衡，提升反硝化除磷系统的脱氮除磷性能.通过长期运行，考察系统在不同模式下的脱氮除磷性能，并进行相应优化;探究不同的操作模式下的脱氮除磷路径，明确各阶段对脱氮除磷的作用;分析不同操作模式下EPS的变化情况，评估新模式对系统稳定性的影响;分析不同操作模式对主要PAOs、DPAOs和DGAOs菌群的调控作用，以期为反硝化除磷系统性能提升提供参考.
1　材料与方法
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1.1　接种污泥与实验用水
接种污泥为课题组中试A/O沉淀池中的污泥与北京高碑店污水处理厂二沉池混合污泥的混合物.反应器厌氧进水为3L的合成生活污水.其组成包括CH3CH2COONa、NH4Cl、KH2PO4、无水CaCl2、MgSO4·7H2O和1mL/L的微量元素[7].采用NaHCO3模拟生活污水的碱度以稳定pH值.具体水质指标为COD:210~290mg/L，NH4+-N:40~60mg/L，PO43--P:6~8mg/L，NO2--N:<1mg/L，NO3－-N:0~2mg/L，pH值为7.5~8.0.此外，在启动期采用3L初始浓度约为30mg/L NO3—-N的NaNO3溶液在A1段开始前进水.
1.2　实验装置与运行方式
为了对比不同的操作模式对系统性能的影响，设3套由有机玻璃制成的单级序批式反应器(SBR).编号R1、R2、R3，每个反应器高径比相同(高度/内径=3)，有效容积为8L，换水比为3/4.3个反应器均设置4个阶段的运行来逐步优化系统，提升系统的脱氮除磷性能，运行周期如图1所示.
为了分析A1/O/A2的最佳比例，在100d的试验中对每个运行周期共480min的周期时长进行了分配.其中进水10min，厌氧120~180min，缺氧进水10min，A1段0~80min(R1的A1段为0min，缺氧进水在O段前)，O段60~120min，好氧沉淀25min，好氧出水10min，A2段40~120min，沉淀25min，排水10min，空闲时间5~90min，不同反应器的具体时间分配如表1所示.在第1阶段(1~50d)，厌氧和缺氧阶段(A1)开始前分别加入3L合成生活废水和3L的NaNO3溶液.将3个SBR反应器的A1/A2时间比分别设置为0、1:2和2:1.第2阶段(51~60d)和第3阶段(61~70d)为过渡期.第2阶段将3个反应器的A1/A2时间比改为上阶段实验得出的最佳A1/A2时间比.在第3阶段在3个反应器的A1段停止加入NaNO3溶液，而是将好氧阶段结束后的出水作为回流储存，然后在下一个循环的A1段开始时流入反应器.在第4阶段(71~100d)，缩短有氧时间以进一步抑制PAOs.
1.3　分析项目与检测方法
本实验包括长期的水质监测和批次试验，批次试验在第48d时进行，除了单个周期的氮、磷和COD的含量外，还对R2的内碳源进行了测定.NH4+-N的测定采用纳氏试剂光度法；NO2－-N的测定采用N-(1-萘基)-乙二胺光度法；NO3－-N的测定采用麝香草酚紫外分光光度法；COD和总磷(TP)的测定采用5B-3B型COD多参数快速测定仪.溶解氧(DO)和pH值的测定采用WTW-Multi3430分析仪.此外，混合液悬浮固体浓度(MLSS)和混合液挥发性悬浮固体浓度(MLVSS)等指标均采用国家规定的标准测量方法[8].
1.4　胞外聚合物(EPS)的分析
EPS测定主要分为3步：EPS提取、蛋白质(PN)测定和多糖(PS)测定.EPS采用改良的热提取法[9]：取30mL泥水混合物，4000g离心5min，收集上清液为溶解性EPS(S-EPS);用70℃的0.05%NaCl溶液恢复体积后，4000g离心10min，收集上清液为松散结合EPS(LB-EPS);再用常温的0.05%NaCl溶液恢复体积后，60℃水浴加热30min,4000g离心15min，收集上清液为紧密结合EPS(TB-EPS).
分别测定S-EPS、LB-EPS及TB-EPS中PN和PS含量，PS测定采用Lowry法，PN测定采用蒽酮硫酸法[10].每个阶段的污泥样品被冷冻干燥，采用Carvalho等[11]和Oehmen等[12]的方法测定PHAs和Gly，其中PHAs的测量基于聚β-羟基丁酸酯(PHB)和聚β-羟基戊酸酯(PHV)的总量.
采用三维荧光光谱法(F97Pro，冷光，中国)对分离后EPS内的有机物组成进一步分析，扫描激发和发射光谱范围均为200~700nm.此外，采用MatLab R2018a(MathWorks Inc.，USA)平台中DOMFluor工具箱进行平行因子(PARAFAC)分析[13]，得到EPS组分的变化.
1.5　微生物组成分析
为了探寻不同操作模式下的微生物群落动态变化规律，在3个反应器运行状态相对稳定的第48d进行取样并根据测序结果进行了微生物群落分析.特别是针对16S区域，由Magigene生物技术有限公司(广州，中国)进行，对细菌结构域进行扩增，重点扩增V3-V4片段，根据Mag-Bind土壤DNA试剂盒说明书进行DNA提取.细菌聚合酶链反应(PCR)扩增采用引物341F(CCTACGGGGNGGCWGCAG)和805R(GACTACHVGGGGTATCTAATCC)[14]，得到的结果使用Illumina MiSeq系统(Illumina MiSeq 2x300bp，USA)进行高通量测序，这些非重复序列以97%的相似阈值进行OTU聚类.
2　结果与讨论
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2.1　不同运行阶段下系统的脱氮除磷性能
根据表1所述，3个反应器在不同操作模式下运行了100d，如图2所示，第1阶段(1~50d)，为了快速恢复污泥活性和增强细胞内碳源储量，反应器设置较长的厌氧段(180min)和好氧段(120min).在R1反应器中，好氧末的PO43－-P浓度由第1d的3.79mg/L降至第22d的1.61mg/L，这可能是因为PAOs活性的增强.此时厌氧末的PO43－-P浓度稳定在(23.61±1.70)mg/L.在第46d，好氧末PO43－-P的浓度稳定在3.49mg/L左右，总出水PO43－-P浓度为(1.09±0.08)mg/L.磷的去除主要发生在好氧段.而NO3－-N的出水浓度下降至稳定在(22.64±0.58)mg/L，这可能是由于DPAOs的缓慢富集.以A/O/A模式运行的R1在46d内初步富集到起除磷作用的PAOs和DPAOs，其中PAOs占主导地位.在R2和R3中，厌氧末磷的含量分别稳定在了(23.85±1.89)mg/L和(23.73±3.16)mg/L.而A1末PO43－-P稳定较早，分别在38d和32d时稳定在(3.12±0.17)mg/L和(2.17±0.10)mg/L.这表明在A/A1/O/A2模式下，R2和R3中磷的去除主要发生在A1段，DPAOs可能为主要功能菌，且R2和R3比R1更早的富集到DPAOs.稳定后的R2反应器的出水NO3－-N浓度(12.69±0.78)mg/L高于R3(9.22±0.35)mg/L，但低于R1(22.64±0.58)mg/L.内源性反硝化可能发生在R2和R3中，DGAOs可能与DPAOs一起富集.此外，第一阶段稳定后R1的氮去除率(NRE)为(67.70%±0.93%)，显著低于R2和R3的(80.31%±1.28%)和(85.06%±0.76%)，这表明采用A/A1/O/A2模式的R2和R3在系统的脱氮性能上相对于R1更有优势.3个反应器的磷去除率(PRE)分别为(85.49%%±0.49%)、(91.52%±0.94%)和(90.56%±1.08%)，R1虽初步富集到了起除磷作用的PAOs和DPAOs两种菌，但在除磷性能上仍稍逊于R2和R3，这可能是由于A1阶段对PAOs的抑制作用.
第2阶段(51~60d)调整R1和R3的A1/A2比值，第3阶段(61~70d)停止加入NaNO3溶液，增设回流装置，主要目的是使NO3－-N和NO2－-N作为底物回流到A1段，供给DPAOs，抑制PAOs，实现整个系统氮的良性循环.加入回流后，R2中NO3－-N浓度显著下降.但A1末的PO43－-P浓度略有上升，达到(3.79±0.05)mg/L，这可能是由于回流中的NO2－-N和NO3－-N浓度(分别为(2.56±0.13)mg/L和(13.61±0.58)mg/L)低于添加的NaNO3溶液浓度(约30mg/L)，一定程度上减弱了DPAOs活性.在第4阶段缩短好氧时间，提高DPAOs的活性.在第4阶段(71~100d)，缩短R1的好氧时间至60min，回流水中含有(5.35±0.39)mg/L的NH4+-N，说明NH4+-N未被完全氧化，好氧时间过短导致出水NH4+-N浓度较高(5.25±0.38)mg/L.A1结束时，PO43--P增加到(5.21±0.17)mg/L，表明反硝化除磷能力也在减弱.R3反应器的好氧段时间保持不变，A1末PO43－-P浓度维持在(3.86±0.13)mg/L，而PRE下降至(86.51%±1.51%)，为增设回流装置减弱除磷菌群的活性提供了支持.在R2中，A1末PO43－-P浓度接近出水，PRE和NRE分别从(91.52%±0.94%)和(80.31%±1.28%)提高至(94.70%±0.96%)和(95.13%±0.35%)，结合R1缩短好氧时间至60min导致NH4+-N未被完全氧化和R3增设回流装置后PRE下降来看，缩短好氧时间至90min既可使NH4+-N反应完全，又有助于进一步提高PRE，抵消回流对除磷造成的负面影响.
与R1相比，采用A/A1/O/A2操作模式的R2和R3在缺氧段除磷效率更高，这可能富集更多的DPAOs.缩短有氧时间至90min有助于进一步抑制PAOs，提高DPAOs活性，最终NRE和PRE分别达到了(95.13%±0.35%)和(94.70%±0.96%).这一去除效率同王文琪等[15]采用间歇曝气A/O/A工艺后期的NRE、PRE(分别为91.15%和95.66%)相比提高了氮的去除效率.与Mandel等[16]在以A/A/O模式运行下通过延长缺氧时间和缩短好氧时间刺激DPAOs的生长使PRE最高达到90.4%的方法相比，对磷的去除效果也更具有优势.
2.2　不同模式下典型周期内污染物去除机理
在第48d的典型周期中监测了3个反应器的物质转化，如图3所示.第48d时R1反应器中大部分的NH4+-N在好氧阶段转化为NO3－-N，少量的NH4+-N被转化为NO2－-N，出水中含有高浓度的NO3－-N，这可能是由于有氧段时间较长导致.在好氧阶段结束时PO43－-P从11.35mg/L降至3.47mg/L，表明R1中磷的去除主要依赖于好氧除磷.在A2段PO43－-P进一步减少，出水浓度仅为1.06mg/L.NO3－-N的浓度从31.03mg/L下降到27.56mg/L，表明体系中存在一定程度的缺氧除磷，但比例较低.缺氧段前的好氧段提供了丰富的氧电子受体，这可能导致了较高的PAOs活性和较低的DPAOs活性.系统主要依靠好氧除磷，反硝化除磷性能不佳.
在R2和R3反应器中，A1段PO43－-P的浓度显著下降，分别从12.84和12.34mg/L降至3.03和2.14mg/L.而NO3－-N水平分别从15.14和15.23mg/L下降到6.35和0.32mg/L.表明在R2和R3中除磷主要发生在稳定后的A1段，先缺氧后好氧的A/A1/O/A2操作模式使得反硝化除磷性能显著加强，从而一定程度上抑制了好氧除磷，DPAOs活性可能得到加强.过度延长A1段时间对除磷的额外影响微乎其微.在好氧段，随着NH4+-N被氧化，R2和R3的NO2－-N和NO3－-N在90min后均趋于稳定(为后续进一步调整O段时间提供了参考)，PO43－-P出水浓度进一步降低，表明A/A1/O/A2操作模式下也存在好氧除磷，但占比较低.在A2段中，NO2－-N和NO3－-N水平进一步降低，猜测可能是由于经常与DPAOs一同出现的DGAOs的内源反硝化导致的.由于A2段进水中含有的高水平NO3－-N在R2和R3中无法完全转化，出水的硝氮仍然较高(12.43和9.12mg/L).COD和PO43－-P的变化在厌氧段的前1.5h最为显著，特别是在前0.5h，这与以往的研究一致[17]，为A2段DGAOs通过内源性代谢去除氮提供了支持.
为进一步了解A/A1/O/A2模式下系统硝化、内源反硝化和除磷的内在机理，在第48d对R2中的胞内碳源(能量来源)进行分析，如图4所示.在A段，PHAs的浓度上升到15.9mmolC/L，Gly浓度下降到10.6mmolC/L，∆PHA/∆COD比值为1.60molC/molC，高于PAOs的1.33molC/molC[18]的理论代谢模型，但低于GAOs的1.86molC/molC[19]模型.这些结果表明PAOs和GAOs都参与了细胞内有机物的转化.如图3，A1段PO43－-P的浓度从12.34mg/L降低到3.53mg/L.PHAs浓度从7.9mmolC/L下降到5.9mmolC/L，Gly浓度从5.3mmolC/L下降到4.5mmolC/L.吸磷量与∆PHA之比(PUA/∆PHA)为0.17molP/molC，接近0.15molP/molC的DPAOs代谢模型[20].∆Gly/∆PHA的比值为0.40molC/molC，介于DPAOs模型的0.30molC/molC和DGAOs模型的0.79molC/molC之间[21].上述结果表明，A1段主要是DPAOs进行反硝化除磷，DGAOs的内源性反硝化作用较小.在A2段Gly浓度从6.9mmolC/L下降到5.7mmolC/L，PHAs浓度从5.0mmolC/L下降到3.2mmolC/L.∆Gly/∆PHA的比值为0.67molC/molC，介于硝酸盐型DGAOs(0.62molC/molC)和亚硝酸盐型DGAOs(1.04molC/molC)的代谢模型之间[22].推测A2段该系统在PHAs和Gly共同驱动下实现了内源性反硝化脱氮.观察到R2中∆PHA/∆COD的比值处于PAOs模型和GAOs模型中间，表明存在混合微生物群落，这些生物的共存可以提供代谢灵活性，允许系统适应不同的环境条件和底物的可用性.在A1段中，磷的吸收以及PHAs和Gly浓度的相应变化为DPAOs和DGAOs的代谢途径提供了能量供给.而接近DPAOs模型的PUA/∆PHA比值又表明了DPAOs是该部分除磷的主要驱动因素.在A2段推测是由于内源反硝化作用，Gly浓度和PHAs浓度下降.该系统可能依靠内部储存的碳源(PHAs和Gly)进行反硝化，这在外部底物有限的情况下至关重要.硝酸盐型和亚硝酸盐型DGAOs的可能存在结果表明，该系统可以通过多种途径高效脱氮，进一步增强了其稳定性和适应性.
综上所述，R1主要依靠好氧除磷，而在以A/A1/O/A2模式运行下的R2与R3中，缺氧除磷是磷去除的主要途径.氮的去除可能依赖于硝化和内源反硝化.功能菌主要在厌氧段转化外部碳源为内碳源储存在细胞内，而好氧段前的A1段为DPAOs创造了良好的缺氧环境，DPAOs利用提供的NO2－-N或NO3－-N底物从原始废水中去除了几乎所有的磷和部分氮，实现缺氧除磷的同时抑制了PAOs.在好氧段AOB将原废水中的氨氮氧化，同时被抑制的PAOs进行一定程度的好氧吸磷.氮的去除主要发生在A2段，此时外部底物有限，系统可能依赖于内部储存的碳源来去除残留的氮，DGAOs在PHAs和Gly的共同驱动下实现了内源性反硝化脱氮.
2.3　不同模式下EPS变化分析
细胞外聚合物(EPS)是一种由细菌产生并附着在细胞表面的粘性物质，是活性污泥的主要成分.EPS在微生物聚集、营养保护、细胞保护、代谢和污染物去除等方面发挥着重要作用[23].图5(a)总结了第48d(第1阶段)和第96d(第4阶段)中EPS蛋白质和多糖的变化.作为EPS的主要成分，PN和PS的总含量与EPS组分的含量成正比，其中TB-EPS含量高于LB-EPS和S-EPS，这有利于污泥的稳定性.第一阶段中R1的PN和PS含量分别为71.62和38.26mg/gVSS，R2中PN和PS含量分别为103.42和58.16mg/gVSS，R3中PN和PS含量分别为126.29和58.02mg/gVSS.大多数PN由疏水性氨基酸组成，疏水性的增加可以增强细胞亲和力，促进细胞聚集[24].PS对于维持污泥的结构完整性至关重要，并能适应微生物的粘附，DGAOs的增加会促进PS的过量分泌[25].与R1相比，R2和R3中PN和PS含量的增加表明稳定性增强，这可能是由于操作模式有利于DGAOs和DPAOs的生长.这表明，与R1相比，以A/A1/O/A2模式运行的R2和R3的EPS含量发生了显著变化，污泥聚集性增强.
利用三维荧光结合PARAFAC模型进行组分拆分和残差分析，将所有荧光光谱分解为3个组分.图5(b)显示了3种组分的光谱位置，图5(c)显示了3种组分的光谱强度变化.在这些物质中，组分1代表了酪氨酸，组分2代表可溶性微生物副产物，组分3与腐植酸样物质内的疏水酸有关[26].拆分后的R2和R3中酪氨酸的荧光强度分别为6172.80和6178.13.而R1的荧光强度仅为2754.27，明显低于R2和R3.腐植酸作为EPS的组分之一，主要是一种高负电官能团的高聚物，不利于污泥的絮凝造粒，导致污泥沉降和活性变差.R2和R3中与腐植酸样物质内的疏水酸有关的荧光强度分别为1434.03和1741.29，低于R1中的2663.61.这可能是由于腐植酸对微生物代谢有抑制作用，腐植酸荧光强度的降低反而说明其生物活性较高.造成这些差异的原因可能是A/A1/O/A2模式的变化以及R1与R2和R3的除磷路径的差异.
2.4　微生物群落结构变化分析
利用MiSeq高通量测序平台对3个反应器第48d的优势微生物群落进行鉴定，如图6所示.维恩图(图6(a))说明了3个反应器之间共享的和唯一的OTUs数量.微生物共有的OTU数为949个，说明反应器内微生物群落存在一定的相似性.而R1、R2以及R3中分别有的81,207和496个独特OTU导致污泥内微生物群落发生实质性变化，这说明由于操作模式的变化生物群落结构也发生了切实的变化.
进一步对样品中的微生物群落由门到属进行分类.在门水平上，3个反应器内微生物组成相似，但丰度不同(图6(b)).在不同的反应器中优势菌门包括变形菌门(Proteobacteria)、拟杆菌门(Bacteroidota)、放线杆菌门(Actinobacteriota)和氯弯菌门(Chloroflexi)，占总菌群的83.69%~90.13%.其中，Proteobacteria和Bacteroidota的比例最高，它们与系统中的反硝化除磷菌密切相关.Proteobacteria包括多种去除有机物、氮和磷的功能微生物，是增强型生物除磷系统中的重要物种[27].Bacteroidota和Chloroflexi参与有机物分解成短链脂肪酸，而Actinobacteriota可以合成促进多糖或酚类化合物分解代谢的酶[28].
在属水平上，优势功能菌属的演替与不同操作设置的变化相对应，如图6(c)所示.典型DPAOs属Dechloromonas在R2和R3中的丰度较R1显著增加，分别从2.24%上升到7.61%和7.94%.Dechloromonas在A1阶段对NO2－-N、NO3－-N和PO43－-P的去除起主要作用，显著提高了R2和R3的反硝化除磷能力.Ca.Accumulibacter是大型生物污水处理厂和实验室规模的除磷反应器中的主要PAOs和DPAOs.它可以使用氧、亚硝酸盐和硝酸盐作为除磷的电子受体[29]，它在3个反应器中的相对丰度分别为8.12%、2.73%和2.22%，这表明采用的A/A1/O/A2模式抑制了以O2为电子受体的PAOs的相关菌群.除此之外，Tetrasphaera已被鉴定为PAOs菌属[30]，虽然与Ca.Accumulibacter相比，Tetrasphaera不储存PHAs，但它可以吸收复杂的有机分子，如氨基酸和葡萄糖[31]，在3个反应器中相对丰度分别为1.41%、0.63%和0.75%. Ca.Accumulibacter与Tetrasphaera的趋势同2.1节R1中磷的去除主要发生在O段，R2和R3中磷的去除主要发生在A1段相吻合，进一步验证了A/A1/O/A2模式对PAOs菌群有抑制作用.Ca.compebacter、Defluviicoccus和Propionivibrio属于DGAOs[5].在操作过程中，Ca.Competibacter为优势DGAOs，其丰度分别为1.53%、4.62%和5.28%，与DPAOs的变化趋势一致.系统中的A1段提供了一个低碳、低氧、低磷的环境，有利于DGAOs的内源反硝化，进一步提高了脱氮效率.
总体而言，微生物群落分析表明A/A1/O/A2操作模式成功地为不同功能微生物创造了协同环境，显著提高了Dechloromonas、Ca.Accumulibacter等DPAOs菌属和Ca.compebacter、Defluviicoccus、Propionivibrio等DGAOs菌属的丰度，在新型DPR系统中获得了良好的氮和磷去除效果.该方法不仅可以最大限度地提高DPAOs和DGAOs的平衡，还可以保证新型DPR系统的稳定性和效率.
3　结论
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3.1　与A/O/A模式相比，A/A1/O/A2模式为不同功能微生物创造了协同环境，通过对系统增设回流装置和缩短有氧段时间的优化，最终NRE和PRE分别达到(95.13%±0.35%)和(94.70%±0.96%)，实现了良好的脱氮除磷效果.
3.2　根据结果推测DPAOs主要在A1段进行反硝化除磷.DGAOs可能在A2段依靠内部储存的碳源(PHAs和Gly)进行内源性反硝化去除残余的氮.
3.3　EPS结果分析表明，相较于以A/O/A模式运行的R1，以A/A1/O/A2模式运行的R2中PN和PS含量分别提高了35.38和12.39mg/gVSS，污泥聚集性得到增强.
3.4　微生物群落结构分析表明，Dechloromonas的丰度从R1的2.24%上升到R2的7.61%和R3的4.62%，Ca.Competibacter的丰度从R1的1.53%上升到R2的7.94%和R3的5.28%，采用A/A1/O/A2模式显著提高了主要DPAOs菌属和主要DGAOs菌属的丰度.
基金
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  • 北京高校卓越青年科学家计划项目(BJJWZYJH01201910005019)
文献
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参考文献 引证文献
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2025年第45卷第6期
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  • 接收时间：2024-11-08
  • 首发时间：2026-02-27
  • 出版时间：2025-06-20
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  • 收稿日期：2024-11-08
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北京高校卓越青年科学家计划项目(BJJWZYJH01201910005019)
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    1.北京工业大学，水质科学与水环境恢复工程北京市重点实验室，北京 100124
    2.哈尔滨工业大学，城市水资源与水环境国家重点实验室，黑龙江 哈尔滨 150090

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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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