Progress of complete ammonia oxidization throughout the wastewater treatment system
HE Zi-xuan1, CHEN Shi1, XU Yi-feng1, LIANG Chuan-zhou1, PENG Lai1,2
1. Hubei Key Laboratory of Mineral Resources Processing and Environment, Wuhan University of Technology, Wuhan 430070, China; 2. Shenzhen Research Institute of Wuhan University of Technology, Shenzhen 518000, China
Abstract:The discovery of complete ammonia-oxidizing bacteria (comammox) provided a novel direction for improving nitrification efficiency in wastewater treatment systems. The potential of their metabolic pathways and functional genes for efficient nitrogen an d carbon removal from wastewater was demonstrated. However, strategies to achieve robust comammox enrichment remained controversial. Further investigations were required to characterize the specific contributions to ammonia removal and nitrous oxide (N2O) production during the nitrification process. The technical strategies for efficient comammox enrichment and the impacts of key factors including environmental substrate concentration, dissolved oxygen, operational processes, and temperature on the selective enrichment of comammox were summarized. Chlorate was employed as a specific inhibitor targeting comammox in combination with 1-octyne to construct a dual-inhibitor experimental system, which was capable of clarifying the nitrification contribution and N2O emission potential of comammox in wastewater treatment systems.
[1] Massara T M, Malamis S, Guisasola A, et al. A review on nitrous oxide (N2O) emissions during biological nutrient removal from municipal wastewater and sludge reject water [J]. Science of the Total Environment, 2017,596-597:106-123. [2] 徐建宇,毛艳萍.从典型硝化细菌到全程氨氧化微生物:发现及研究进展[J]. 微生物学通报, 2019,46(4):879-890. Xu J, Mao Y. From canonical nitrite oxidizing bacteria to complete ammoniaoxidizer: discovery and advances [J]. Microbiology China, 2019,46(4):879-890. [3] Costa E, Pérez J, Kreft J U. Why is metabolic labour divided in nitrification? [J]. Trends in Microbiology, 2006,14(5):213-219. [4] Daims H, Lebedeva E V, Pjevac P, et al. Complete nitrification by Nitrospira bacteria [J]. Nature, 2015,(7583):504-509. [5] Van Kessel M A H J V, Speth D R, Albertsen M, et al. Complete nitrification by a single microorganism [J]. Nature, 2015,528(7583): 555-559. [6] Kits K D, Sedlacek C J, Han P, et al. Kinetic analysis of a complete nitrifier reveals an oligotrophic lifestyle [J]. Nature, 2017,549(7671): 269-272. [7] Xu Y, Liu G, Hua Y, et al. The diversity of comammox bacteria and the effect of sewage discharge on their abundance in eutrophic lake sediments [J]. Journal of Soils and Sediments, 2020,20(5):2495-2503. [8] Ye J, Zhao S, Ren J, et al. Higher contribution by comammox bacteria than AOA and AOB to nitrification in the sediments of lake Taihu [J]. International Biodeterioration & Biodegradation, 2024,187:105709. [9] Sun D, Tang X, Li J, et al. Chlorate as a comammox Nitrospira specific inhibitor reveals nitrification and N2O production activity in coastal wetland [J]. Soil Biology and Biochemistry, 2022,173:108782. [10] Zhang A, Zhu M, Zheng Y, et al. The significant contribution of comammox bacteria to nitrification in a constructed wetland revealed by DNA-based stable isotope probing [J]. Bioresource Technology, 2024,399:130637. [11] Li C, Hu H, Chen Q, et al. Comammox nitrospira play an active role in nitrification of agricultural soils amended with nitrogen fertilizers [J]. Soil Biology & Biochemistry, 2019,138:107609. [12] Wagner F B, Diwan V, Dechesne A, et al. Copper-induced stimulation of nitrification in biological rapid sand filters for drinking water production by proliferation of nitrosomonas spp [J]. Environmental Science & Technology, 2019,53(21):12433-12441. [13] Wang Z, Cao Y, Zhu-Barker X, et al. Comammox nitrospira clade B contributes to nitrification in soil [J]. Soil Biology & Biochemistry, 2019,135:392-395. [14] Wang X, Wang S, Jiang Y, et al. Comammox bacterial abundance, activity, and contribution in agricultural rhizosphere soils [J]. Science of the Total Environment, 2020,727:138563. [15] Tan C, Yin C, Zhang L, et al. Deciphering the functional importance of comammox vs. canonical ammonia oxidisers in nitrification and N2O emissions in acidic agricultural soils [J]. Soil Biology and Biochemistry, 2024,193:109415. [16] Zhang K, Sun H, Kang Y, et al. Comammox plays a functionally important role in the nitrification of rice paddy soil with different nitrogen fertilization levels [J]. Applied Soil Ecology, 2024,193: 105120. [17] How S W, Nittami T, Ngoh G C, et al. An efficient oxic-anoxic process for treating low COD/N tropical wastewater: Startup, optimization and nitrifying community structure [J]. Chemosphere, 2020,259:127444. [18] Zheng M, Wang M, Zhao Z, et al. Transcriptional activity and diversity of comammox bacteria as a previously overlooked ammonia oxidizing prokaryote in full-scale wastewater treatment plants [J]. Science of the Total Environment, 2019,656(1):717-722. [19] He S, Zhao Z, Tian Z, et al. Comammox bacteria predominate among ammonia-oxidizing microorganisms in municipal but not in refinery wastewater treatment plants [J]. Journal of Environmental Management, 2022,316:115271. [20] Wang M, Huang G, Zhao Z, et al. Newly designed primer pair revealed dominant and diverse comammox amoA gene in full_scale wastewater treatment plants [J]. Bioresource Technology, 2018,270(1):580-587. [21] Pinto A J, Marcus D N, Ijaz U Z, et al. Metagenomic evidence for the presence of comammox nitrospira-like bacteria in a drinking water system [J]. mSphere, 2016,1(1):e00054-00015. [22] Hu J, Zhao Y, Yang W, et al. Surface ammonium loading rate shifts ammonia-oxidizing communities in surface water-fed rapid sand filters [J]. Fems Microbiology Ecology, 2020,96(10):9. [23] Wang Y, Ma L, Mao Y, et al. Comammox in drinking water systems [J]. Water Research, 2017,116:332-341. [24] Ben M, Timothy M L, Taegyu K, et al. Multi-scale investigation of ammonia-oxidizing microorganisms in biofilters used for drinking water treatment [J]. Environmental Science & Technology, 2023,57(9): 3833-3842. [25] Tatari K, Uuml, Lay A, et al. Challenges in using allylthiourea and chlorate as specific nitrification inhibitors [J]. Chemosphere, 2017,182: 301-305. [26] Liu S, Cai H, Wang J, et al. In-situ expressions of comammox Nitrospira along the Yangtze River [J]. Water Research, 2021,200: 117241. [27] Wang D, Zhou C, Nie M, et al. Abundance and niche specificity of different types of complete ammonia oxidizers (comammox) in salt marshes covered by different plants [J]. Science of the Total Environment, 2021,768:144993. [28] Xia F, Gao J, Wang J, et al. Ubiquity and diversity of complete ammonia oxidizers (comammox) [J]. Applied and Environemental Microbiology, 2018,84(24):e01390-01318. [29] Zhao Z, Huang G, He S, et al. Abundance and community composition of comammox bacteria in different ecosystems by a universal primer set [J]. Science of the Total Environment, 2019,691:146-155. [30] Sakoula D, Koch H, Frank J, et al. Enrichment and physiological characterization of a novel comammox Nitrospira indicates ammonium inhibition of complete nitrification [J]. ISME Journal, 2021,15(4):1010-1024. [31] 孙东耀.河口潮滩湿地全程硝化菌群落特征及其生态功能研究[D]. 上海:华东师范大学, 2022. Sun D. Distribution pattern and ecological function of Comammox Nitrospira bacteria in estuarine tidal flat wetlands [D]. Shanghai: East China Normal University, 2022. [32] Wang Y, Zhao R, Liu L, et al. Selective enrichment of comammox from activated sludge using antibiotics [J]. Water Research, 2021, 197:117087. [33] Gonzalez-Martinez A, Rodriguez-Sanchez A, Loosdrecht M, et al. Detection of comammox bacteria in full-scale wastewater treatment bioreactors using tag-454-pyrosequencing [J]. Environmental Science and Pollution Research, 2016,23(24):25501-25511. [34] Pjevac P, Schauberger C, Poghosyan L, et al. AmoA-targeted polymerase chain reaction primers for the specific detection and quantification of comammox nitrospira in the environment [J]. Frontiers in Microbiology, 2017,8:1508. [35] Wang M, Huang G, Zhao Z, et al. Newly designed primer pair revealed dominant and diverse comammox amoA gene in full-scale wastewater treatment plants [J]. Bioresource Technology, 2018,270:580-587. [36] Yang Y, Daims H, Liu Y, et al. Activity and metabolic versatility of complete ammonia oxidizers in full-scale wastewater treatment systems [J]. Mbio, 2020,11(2):e03175-19. [37] Spasov E, Tsuji J M, Hug L A, et al. High functional diversity among Nitrospira populations that dominate rotating biological contactor microbial communities in a municipal wastewater treatment plant [J]. ISME Journal, 2020,14(7):1857-1872. [38] Tsuchiya Y, Nakagawa T, Takahashi R. Quantification and phylogenetic analysis of ammonia oxidizers on biofilm carriers in a full-scale wastewater treatment plant [J]. Microbes and Environments, 2020,35(2):ME19140. [39] Roots P, Wang Y, Rosenthal A F, et al. Comammox nitrospira are the dominant ammonia oxidizers in a mainstream low dissolved oxygen nitrification reactor [J]. Water Research, 2019,157:396-405. [40] Li J, Hua Z, Liu T, et al. Selective enrichment and metagenomic analysis of three novel comammox Nitrospira in a urine-fed membrane bioreactor [J]. ISME Communications, 2021,1(1):7-7. [41] Fujitani H, Nomachi M, Takahashi Y, et al. Successful enrichment of low-abundant comammox Nitrospira from nitrifying granules under ammonia-limited conditions [J]. Fems Microbiology Letters, 2020, 367(1). [42] Cui H, Zhang L, Zhang Q, et al. Enrichment of comammox bacteria in anammox-dominated low-strength wastewater treatment system within microaerobic conditions: Cooperative effect driving enhanced nitrogen removal [J]. Chemical Engineering Journal, 2023,453: 139851. [43] Shao Y, Wu J, Chen H. Comammox nitrospira cooperate with anammox bacteria in a partial nitritation - anammox membrane bioreactor treating low-strength ammonium wastewater at high loadings [J]. Water Research, 2024,257:121698. [44] Huang T, Xia J, Liu T, et al. Comammox nitrospira bacteria are dominant ammonia oxidizers in mainstream nitrification bioreactors emended with sponge carriers [J]. Environmental Science & Technology, 2022,56(17):12584-12591. [45] Zhao Y, Wang J, Liu Z, et al. Biofilm: A strategy for the dominance of comammox Nitrospira [J]. Journal of Cleaner Production, 2022, 363:132361. [46] Guo Z, Ma X, El-Baz A, et al. Revolutionizing comammox enrichment: Novel approaches for comammox enrichment through reactor configuration and ammonia concentration mediation [J]. Journal of Water Process Engineering, 2024,60:105172. [47] Hou J, Zhu Y, Liu J, et al. Competitive enrichment of comammox Nitrospira in floccular sludge [J]. Water Research, 2024,251:121151. [48] Camejo P Y, Domingo J S, Mcmahon K D, et al. Genome-enabled insights into the ecophysiology of the comammox bacterium "Candidatus Nitrospira nitrosa" [J]. Msystems, 2017,2(5):e00059-17. [49] Zhao Y, Hu J, Yang W, et al. The long-term effects of using nitrite and urea on the enrichment of comammox bacteria [J]. Science of the Total Environment, 2021,755(2):142580. [50] Zhao J, Zheng M, Su Z, et al. Selective enrichment of comammox nitrospira in a moving bed biofilm reactor with sufficient oxygen supply [J]. Environmental Science & Technology, 2022,56(18): 13338-13346. [51] Koch H, Van-Kessel M, Lücker S, et al. Complete nitrification: insights into the ecophysiology of comammox nitrospira [J]. Applied Microbiology & Biotechnology, 2019,103(1):177-189. [52] Zhang H, Cheng F, Sun S, et al. Diversity distribution and characteristics of comammox in different ecosystems [J]. Environmental Research, 2022,214:113900. [53] 胡万超,赵琛,王巧娟,等.饮用水快速砂滤池优势微生物群落的代谢功能解析[J]. 环境科学, 2019,40(8):3604-3611. Hu W, Zhao C, Wang Q, et al. Metabolic functional analysis of dominant microbial communities in the rapidSand filters for drinking water [J]. Environmental Science, 2019,40(8):3604-3611. [54] Emilie S, Jackson M T, Laura A H, et al. High functional diversity among Nitrospira populations that dominate rotating biological contactor microbial communities in a municipal wastewater treatment plant [J]. The ISME Journal, 2020,14(7):1857-1872. [55] Guo Z, Ma X, El-Baz A, et al. Revolutionizing comammox enrichment: Novel approaches for comammox enrichment through reactor configuration and ammonia concentration mediation [J]. Journal of Water Process Engineering, 2024,60:105172. [56] Li J, Hua Z, Liu T, et al. Selective enrichment and metagenomic analysis of three novel comammox Nitrospira in a urine-fed membrane bioreactor [J]. ISME Communications, 2021,1(1):7. [57] Zhao Y, Hu J, Yang W, et al. The long-term effects of using nitrite and urea on the enrichment of comammox bacteria [J]. Science of the Total Environment, 2021,755:142580. [58] 李旭,周鑫,王共磊,等.全程氨氧化菌(Comammox)在污水生物脱氮中的应用进展[J]. 中国给水排水, 2023,39(22):46-54. Li X, Zhou X, Wang G, et al. Advances in the application of comammox in biological nitrogen removalfrom wastewater [J]. China Water & Wastewater, 2023,39(22):46-54. [59] 张亮,于静仪,李朝阳,等.污水生物处理系统中全程氨氧化菌的研究进展[J]. 北京工业大学学报, 2020,46(4):402-411. Zhang L, Yu J, Li C, et al. Progress and prospects of the research of complete ammonia oxidizers in biological wastewater treatment [J]. Journal of Beijing University of Technology, 2020,46(4):402-411. [60] Zhang S, Wang J, Wang D, et al. Abundance and niche differentiation of comammox in the sludges of wastewater treatment plants that use the anaerobic-anoxic-aerobic process [J]. Life-Basel, 2022,12(7):954. [61] Dimitra S, Hanna K, Jeroen F, et al. Enrichment and physiological characterization of a novel comammox Nitrospira indicates ammonium inhibition of complete nitrification [J]. The ISME Journal, 2021,15(4):1010-1024. [62] Hou J, Zhu Y, Liu J, et al. Competitive enrichment of comammox Nitrospira in floccular sludge [J]. Water Research, 2024,251:121151. [63] Cui H, Zhang L, Zhang Q, et al. Enrichment of comammox bacteria in anammox-dominated low-strength wastewater treatment system within microaerobic conditions: Cooperative effect driving enhanced nitrogen removal [J]. Chemical Engineering Journal, 2023,453: 139851. [64] Zheng M, Mu G, Zhang A, et al. Predominance of comammox bacteria among ammonia oxidizers under low dissolved oxygen condition [J]. Chemosphere, 2022,308:136436. [65] Zhao J, Zheng M, Su Z, et al. Selective enrichment of comammox nitrospira in a moving bed biofilm reactor with sufficient oxygen supply [J]. Environmental Science & Technology, 2022,56(18): 13338-13346. [66] 池玉蕾,石烜,任童,等.溶解氧对低碳源城市污水处理系统脱氮性能与微生物群落的影响[J]. 环境科学, 2021,42(9):4374-4382. Chi Y, Shi X, Ren T, et al. Effects of dissolved oxygen o [67] 赖永恒,刘敏.A/O-MBBR工艺处理生活污水性能及菌群结构[J]. 中国环境科学, 2022,42(5):2120-2128. Lai Y, Liu M. Performance and microbial community of anoxic/ oxicmoving bed biofilm reactor(A/O-MBBR) process treating domesticsewage. [J]. China Environmental Science, 2022,42(5):2120-2128. [68] Zhu Y, Hou J, Liu J, et al. Model-based development of strategies enabling effective enrichment and application of comammox bacteria in floccular sludge under mainstream conditions [J]. Science of the Total Environment, 2023,895:165051. [69] Irmarie C, Zihan D, Linxuan H, et al. Long solids retention times and attached growth phase favor prevalence of comammox bacteria in nitrogen removal systems [J]. Water Research, 2020,169:115268. [70] Wang D, Wang Y, Liu L, et al. Niche differentiation and symbiotic association among ammonia/nitrite oxidizers in a full-scale rotating biological contactor [J]. Water Research, 2022,225:119137. [71] Zhou X, Liu H, Fan X, et al. Comparative analysis of bacterial information of biofilms and activated sludge in full-scale MBBR-IFAS systems [J]. Microorganisms, 2024,12(6):1121. [72] Huang T, Xia J, Liu T, et al. Comammox nitrospira bacteria are dominant ammonia oxidizers in mainstream nitrification bioreactors emended with sponge carriers [J]. Environmental science & technology, 2022,56(17):12584-12591. [73] Sotto R D, Bae S. Nutrient removal performance and microbiome of an energy-efficient reciprocation MLE-MBR operated under hypoxic conditions [J]. Water Research, 2020,182:115991. [74] How S W, Nittami T, Ngoh G C, et al. An efficient oxic-anoxic process for treating low COD/N tropical wastewater: Startup, optimization and nitrifying community structure [J]. Chemosphere, 2020,259:127444. [75] Li H, Zhang Y, Yang M, et al. Effects of hydraulic retention time on nitrification activities and population dynamics of a conventional activated sludge system [J]. Frontiers of Environmental Science & Engineering, 2013,7(1):43-48. [76] Zhao Y, Wang J, Liu Z, et al. Biofilm: A strategy for the dominance of comammox Nitrospira [J]. Journal of Cleaner Production, 2022,363: 132361. [77] Fowler S J, Palomo A, Dechesne A, et al. Comammox nitrospira are abundant ammonia oxidizers in diverse groundwater-fed rapid sand filter communities [J]. Environmental Microbiology, 2018,20(3):1002-1015. [78] Zhou X, Li B L, Wei J, et al. Temperature influenced the comammox community composition in drinking water and wastewater treatment plants [J]. Microbial Ecology, 2021,82(4):870-884. [79] Pan K, Gao J, Fan X, et al. The more important role of archaea than bacteria in nitrification of wastewater treatment plants in cold season despite their numerical relationships [J]. Water Research, 2018,145:552-561. [80] Li B, Godfrey B J, Redcorn R, et al. Mainstream nitrogen removal from low temperature and low ammonium strength municipal wastewater using hydrogel-encapsulated comammox and anammox [J]. Water Research, 2023,242:120303. [81] Massara T M, Malamis S, Guisasola A, et al. A review on nitrous oxide (N2O) emissions during biological nutrient removal from municipal wastewater and sludge reject water [J]. Science of the Total Environment, 2017,596:106-123. [82] Ni B, Yuan Z. Recent advances in mathematical modeling of nitrous oxides emissions from wastewater treatment processes [J]. Water Research, 2015,87:336-346. [83] Mehrani M J, Lu X, Kowal P, et al. Incorporation of the complete ammonia oxidation (comammox) process for modeling nitrification in suspended growth wastewater treatment systems [J]. Journal of Environmental Management, 2021,297:113223. [84] Yang X, Yu X, He Q, et al. Niche differentiation among comammox (Nitrospira inopinata) and other metabolically distinct nitrifiers [J]. Frontiers in Microbiology, 2022,13:956860. [85] Mehrani M J, Kowal P, Sobotka D, et al. The coexistence and competition of canonical and comammox nitrite oxidizing bacteria in a nitrifying activated sludge system-Experimental observations and simulation studies [J]. Science of the Total Environment, 2023,864:161084. [86] Mei P, Wang Z, Guo W, et al. The ASM2d model with two-step nitrification can better simulate biological nutrient removal systems enriched with complete ammonia oxidizing bacteria (comammox Nitrospira) [J]. Chemosphere, 2023,335:139169. [87] Al-Ajeel S, Spasov E, Sauder L A, et al. Ammonia-oxidizing archaea and complete ammonia-oxidizing Nitrospira in water treatment systems [J]. Water Research X, 2022,15:100131. [88] Chao Y, Mao Y, Yu K, et al. Novel nitrifiers and comammox in a full-scale hybrid biofilm and activated sludge reactor revealed by metagenomic approach [J]. Applied Microbiology and Biotechnology, 2016,100(18):8225-8237. [89] 余杰.全程氨氧化菌的分布、活性及N2O释放的影响研究——以典型白洋淀岸边带为例[D]. 无锡:江南大学, 2023. Yu J. Distribution, activity and N2O production of Comammox: take a typical Baiyangdian riparian zone as an example [D]. Wuxi: Jiangnan University, 2023. [90] Metch J W, Wang H, Ma Y J, et al. Insights gained into activated sludge nitrification through structural and functional profiling of microbial community response to starvation stress [J]. Environmental Science-Water Research & Technology, 2019,5(5):884-896. [91] Orellana L H, Hatt J K, Iyer R, et al. Comparing DNA, RNA and protein levels for measuring microbial dynamics in soil microcosms amended with nitrogen fertilizer [J]. Scientific Reports, 2019,9(1):17630. [92] Gülay A, Fowler S J, Tatari K, et al. DNA- and RNA-SIP Reveal Nitrospira spp. as Key Drivers of Nitrification in Groundwater-Fed Biofilters [J]. Mbio, 2019,10(6):e01870-19. [93] Ding X, Lan W, Li Y, et al. An internal recycling mechanism between ammonia/ammonium and nitrate driven by ammonia-oxidizing archaea and bacteria (AOA, AOB, and Comammox) and DNRA on Angkor sandstone monuments [J]. International Biodeterioration & Biodegradation, 2021,165:105328. [94] Maria M, Katharina K, Arno S, et al. Flow-through stable isotope probing (Flow-SIP) minimizes cross-feeding in complex microbial communities [J]. The ISME Journal, 2021,15(1):348-353. [95] 张奇春,巩龙达,牛天新.氨氧化微生物对土壤硝化贡献的测量方法, 202210561194.7[P]. 2022-05-23. Zhang C, Gong L, Niu T. Measurement of the contribution of ammonia-oxidizing microorganisms to soil nitrification, 202210561194.7[P]. 2022-05-23. [96] Zheng M, Tian Z, Chai Z, et al. Ubiquitous occurrence and functional dominance of comammox Nitrospira in full-scale wastewater treatment plants [J]. Water Research, 2023,236:119931. [97] Neufeld J D, Dumont M G, Vohra J, et al. Methodological considerations for the use of stable isotope probing in microbial ecology [J]. Microbial Ecology, 2007,53(3):435-442. [98] Liu S, Han P, Hink L, et al. Abiotic conversion of extracellular NH2OH contributes to N2O emission during ammonia oxidation [J]. Environmental Science & Technology, 2017,51(22):13122-13132. [99] Kits K D, Sedlacek C J, Lebedeva E V, et al. Kinetic analysis of a complete nitrifier reveals an oligotrophic lifestyle [J]. Nature, 2017,549(7671):269-272. [100] Li D, Fang F, Liu G. Efficient nitrification and low-level N2O emission in a weakly acidic bioreactor at low dissolved-oxygen levels are due to comammox [J]. Applied and Environemental Microbiology, 2021,87(11):1-13. [101] Ren Z, Li D, Zhang Z, et al. Enhancing the relative abundance of comammox nitrospira in ammonia oxidizer community decreases N2O emission in nitrification exponentially [J]. Chemosphere, 2024,356:141883. [102] Kits K D, Jung M, Vierheilig J, et al. Low yield and abiotic origin of N2O formed by the complete nitrifier Nitrospira inopinata [J]. Nature Communications, 2019,10(1):1-12. [103] Wan X, Yang X, Zhang Y, et al. Lithology-driven soil properties control of N2O production by ammonia oxidizers in subtropical forest soils [J]. Geoderma, 2024,446:116910. [104] Tan C, Yin C, Li W, et al. Comammox nitrospira play a minor role in N2O emissions from an alkaline arable soil [J]. Soil Biology and Biochemistry, 2022,171:108720. [105] Vasilaki V, Massara T M, Stanchev P, et al. A decade of nitrous oxide (N2O) monitoring in full-scale wastewater treatment processes: A critical review [J]. Water Research, 2019,161:392-412. [106] Chen W, Yang J, Yuan C, et al. Toward better understanding and feasibility of controlling greenhouse gas emissions from treatment of industrial wastewater with activated sludge [J]. Environmental Science and Pollution Research, 2016,23(20):20449-20461. [107] Pan Y, Van Den Akker B, Ye L, et al. Unravelling the spatial variation of nitrous oxide emissions from a step-feed plug-flow full scale wastewater treatment plant [J]. Scientific Reports, 2016,6:20792. [108] Mampaey K E, De Kreuk M K, Van Dongen U, et al. Identifying N2O formation and emissions from a full-scale partial nitritation reactor [J]. Water Research, 2016,88:575-585. [109] Zhu Y, Hou J, Meng F, et al. Role of comammox bacteria in granular bioreactor for nitrogen removal via partial nitritation/anammox [J]. Bioresource Technology, 2024,406:131070. [110] Shao Y, Wu J. Comammox nitrospira species dominate in an efficient partial nitrification-anammox bioreactor for treating ammonium at low loadings [J]. Environmental Science & Technology, 2021,55(3): 2087-2098. [111] Vilardi K, Cotto I, Bachmann M, et al. Co-occurrence and cooperation between comammox and anammox bacteria in a full-scale attached growth municipal wastewater treatment process [J]. Environmental Science & Technology, 2023,57(12):5013-5023. [112] Gottshall E Y, Bryson S J, Cogert K I, et al. Sustained nitrogen loss in a symbiotic association of comammox nitrospira and anammox bacteria [J]. Water Research, 2021,202:117426.n nutrient removal performance and microbial community in low carbon/ nitrogen municipal wastewater treatment process [J]. Environmental Science, 2021,42(9):4374-4382.