Changes in microbial structure in the anammox population at high-temperature

Öz

Anammox bacteria are crucial in converting ammonia nitrogen into N2 gas in anoxic conditions, offering sustainable alternatives to biological wastewater treatment. However, temperature fluctuations in wastewater treatment plants significantly impact the activity of microorganisms, including anammox bacteria. This underscores the importance of understanding the influence of temperature on microbial ecology. Although extensive research has been conducted on anammox activity across various temperatures, the effects of high temperatures on microbial diversity within anammox consortia remain largely unexplored. This study aims to investigate the effects of high temperatures on microbial performance and shifts in microbial diversity in anammox bioreactors. Two separate bioreactors were employed to validate the findings. After enriching anammox cultures under mesophilic conditions, the bioreactors were exposed to 65°C for 9 hours, followed by a recovery phase at 35°C. Thermal stress led to a sharp decline in nitrogen removal efficiency, with NH4+-N and NO2--N removal rates decreasing by up to 83.59% and 91.35%, respectively. Even after reducing influent concentrations, nitrogen removal performance remained impaired. Metagenomic analyses revealed substantial shifts in microbial composition at elevated temperatures. While phyla such as Chloroflexi and Bacteroidota diminished, Proteobacteria and Firmicutes proliferated. Notably, Candidatus Kuenenia (anammox genus) declined, indicating its sensitivity to thermal stress. These findings provide valuable insights into how high temperatures influence microbial diversity in anammox systems. Future research should focus on improving microbial resilience and stability to optimize anammox system performance in harsh environmental conditions.

Anahtar Kelimeler

• Anammox
• Wastewater treatment
• High temperature
• Metagenomics
• Microbial composition

Yüksek sıcaklıkta anammox popülasyonunda mikrobiyal yapıdaki değişiklikler

Öz

Anammox bakterileri, anoksik koşullar altında amonyak azotunu N2 gazına dönüştürmede kritik öneme sahiptir ve biyolojik atık su arıtımına sürdürülebilir alternatifler sunar. Ancak, atık su arıtma tesislerindeki sıcaklık dalgalanmaları, anammox bakterileri de dahil olmak üzere mikroorganizmaların aktivitelerini önemli ölçüde etkilemektedir. Bu durum, sıcaklığın mikrobiyal ekoloji üzerindeki etkisini anlamanın önemini ortaya koymaktadır. Çeşitli sıcaklıklarda anammox aktivitesi üzerine kapsamlı araştırmalar yürütülmüş olmasına rağmen, yüksek sıcaklıkların anammox konsorsiyumlarındaki mikrobiyal çeşitlilik üzerindeki etkileri büyük ölçüde keşfedilmemiştir. Bu çalışma, anammox biyoreaktörlerinde yüksek sıcaklıkların mikrobiyal performans üzerindeki etkilerini ve mikrobiyal çeşitlilikteki değişimleri araştırmayı amaçlamaktadır. Bulguları doğrulamak için iki ayrı biyoreaktör kullanılmıştır. Anammox kültürleri mezofilik koşullar altında zenginleştirildikten sonra, biyoreaktörler 9 saat boyunca 65 °C’ye maruz bırakılmış ve ardından 35 °C’de bir geri kazanım sürecine tabi tutulmuştur. Termal stres, azot giderim verimliliğinde keskin bir düşüşe neden olmuş; NH4+-N ve NO2--N giderim oranları sırasıyla %83,59 ve %91,35'e kadar azalmıştır. Giriş konsantrasyonlarının azaltılmasına rağmen, azot giderim performansı iyileşme göstermemiştir. Metagenomik analizler, yüksek sıcaklıklarda mikrobiyal bileşimde önemli değişiklikler olduğunu ortaya koymuştur. Chloroflexi ve Bacteroidota gibi filumlar azalırken, Proteobakteriler ve Firmicutes artmıştır. Özellikle, Candidatus Kuenenia (anammox türü) bir düşüş göstermiştir ve bu durum bu türün termal strese duyarlılığını ortaya koymaktadır. Bu bulgular, yüksek sıcaklıkların anammox sistemlerindeki mikrobiyal çeşitlilik üzerindeki etkilerine ilişkin değerli bilgiler sunmaktadır. Gelecekteki araştırmalar, zorlu çevre koşullarında anammox sistemlerinin performansını optimize etmek için mikrobiyal dayanıklılığı ve stabiliteyi artırmaya odaklanmalıdır.

Anahtar Kelimeler

• Anammox
• Atık su arıtımı
• Yüksek sıcaklık
• Metagenomik
• Mikrobiyal kompozisyon

Kaynakça

1. [1] Jetten MSM, Strous M, van de Pas-Schoonen KT, Schalk J, van Dongen UGJM, van de Graaf AA, Logemann S, Muyzer G, van Loosdrecht MCM, Kuenen JG. "The anaerobic oxidation of ammonium". FEMS Microbiology Reviews, 22(5), 421-437, 1998.
2. [2] Sonthiphand P, Hall MW, Neufeld JD. "Biogeography of anaerobic ammonia-oxidizing (anammox) bacteria". Frontiers in Microbiology, 5, 2014.
3. [3] Wang P, Lu B, Liu X, Chai X. "Accelerating the granulation of anammox sludge in wastewater treatment with the drive of “micro-nuclei”: A review". Science of the Total Environment, 860, 160238, 2023.
4. [4] Adams M, Issaka E, Chen C. "Anammox-based technologies: A review of recent advances, mechanism, and bottlenecks". Journal of Environmental Sciences, 148, 151-173, 2025.
5. [5] Trinh HP, Lee S-H, Jeong G, Yoon H, Park H-D. "Recent developments of the mainstream anammox processes: Challenges and opportunities". Journal of Environmental Chemical Engineering, 9(4), 105583, 2021.
6. [6] Ren Z-Q, Wang H, Zhang L-G, Du X-N, Huang B-C, Jin RC. "A review of anammox-based nitrogen removal technology: From microbial diversity to engineering applications". Bioresource Technology, 363, 127896, 2022.
7. [7] Wu P, Chen J, Garlapati VK, Zhang X, Wani Victor Jenario F, Li X, Liu W, Chen C, Aminabhavi TM, Zhang X. "Novel insights into anammox-based processes: A critical review". Chemical Engineering Journal, 444, 136534, 2022.
8. [8] Wang Q, Tan G-YA, Azari M, Huang X, Denecke M, Men Y, Jung J-Y, Okabe S, Ali M, Huang Y-T, Wu Z, Lo W-h, Gu JD, Lin J-G, Lee P-H. "Insights into the roles of anammox bacteria in post-treatment of anaerobically-treated sewage". Critical Reviews in Environmental Science and Technology, 48(6), 655-684, 2018.

9. [9] Lackner S, Gilbert EM, Vlaeminck SE, Joss A, Horn H, van Loosdrecht MCM. "Full-scale partial nitritation/anammox experiences – an application survey". Water Research, 55, 292-303, 2014.
10. [10] Zuo F, Yue W, Gui S, Sui Q, Wei Y. "Resilience of anammox application from sidestream to mainstream: A combined system coupling denitrification, partial nitritation and partial denitrification with anammox". Bioresource Technology, 374, 128783, 2023.
11. [11] Huang X, Mi W, Chan YH, Singh S, Zhuang H, Leu S-Y, Li X-z, Li X, Lee P-H. "C-n-s synergy in a pilot-scale mainstream anammox fluidized-bed membrane bioreactor for treating chemically enhanced primary treatment saline sewage". Water Research, 229, 119475, 2023.
12. [12] Cao S, Koch K, Du R, Wells GF, Ye L, Drewes JE. "Toward mainstream anammox by integrating sidestream treatment". Environmental Science & Technology, 56(15), 10553-10556, 2022.
13. [13] Shaw DR, Tobon Gonzalez J, Bibiano Guadarrama C, Saikaly PE. "Emerging biotechnological applications of anaerobic ammonium oxidation". Trends in Biotechnology, 42(9), 1128-1143, 2024.
14. [14] Gonzalez JM, Aranda B. "Microbial growth under limiting conditions-future perspectives". Microorganisms, 11(7), 2023.
15. [15] Byrne N, Strous M, Crépeau V, Kartal B, Birrien J-L, Schmid M, Lesongeur F, Schouten S, Jaeschke A, Jetten M, Prieur D, Godfroy A. "Presence and activity of anaerobic ammonium-oxidizing bacteria at deep-sea hydrothermal vents". The ISME Journal, 3(1), 117-123, 2008.
16. [16] Kouba V, Bachmannová C, Podzimek T, Lipovová P, van Loosdrecht MCM. "Physiology of anammox adaptation to low temperatures and promising biomarkers: A review". Bioresource Technology, 349, 126847, 2022.
17. [17] Tomaszewski M, Cema G, Ziembinska-Buczynska A. "Influence of temperature and ph on the anammox process: A review and meta-analysis". Chemosphere, 182, 203-214, 2017.
18. [18] Zhang L, Okabe S. "Ecological niche differentiation among anammox bacteria". Water Research, 171, 115468, 2020.
19. [19] Strous M, Kuenen JG, Jetten MSM. "Key physiology of anaerobic ammonium oxidation". Applied and Environmental Microbiology, 65(7), 3248-3250, 1999.
20. [20] Le L-T, Jeon J, Dang B-T, Bui X-T, Jahng D. "Influence of temperature on anammox reaction and microbial diversity in a bio-carriers reactor under mainstream conditions". Environmental Technology & Innovation, 25, 102178, 2022.
21. [21] Chen X, Liu L, Bi Y, Meng F, Wang D, Qiu C, Yu J, Wang S. "A review of anammox metabolic response to environmental factors: Characteristics and mechanisms". Environmental Research, 223, 115464, 2023.
22. [22] Lotti T, Kleerebezem R, van Loosdrecht MCM. "Effect of temperature change on anammox activity". Biotechnology and Bioengineering, 112(1), 98-103, 2015.
23. [23] Dosta J, Fernández I, Vázquez-Padín JR, MosqueraCorral A, Campos JL, Mata-Álvarez J, Méndez R. "Short- and long-term effects of temperature on the anammox process". Journal of Hazardous Materials, 154(1), 688- 693, 2008.
24. [24] Han YM. "Study on temperature influence on nitrogen removal properties in anammox". Advanced Materials Research, 955-959, 484-487, 2014.
25. [25] Toh SK, Webb RI, Ashbolt NJ. "Enrichment of autotrophic anaerobic ammonium-oxidizing consortia from various wastewaters". Microbial Ecology, 43(1), 154-167, 2002.
26. [26] Isanta E, Bezerra T, Fernández I, Suárez-Ojeda ME, Pérez J, Carrera J. "Microbial community shifts on an anammox reactor after a temperature shock using 454- pyrosequencing analysis". Bioresource Technology, 181, 207-213, 2015.
27. [27] Shu W-S, Huang L-N. "Microbial diversity in extreme environments". Nature Reviews Microbiology, 20(4), 219- 235, 2022.
28. [28] Li S-J, Hua Z-S, Huang L-N, Li J, Shi S-H, Chen L-X, Kuang J-L, Liu J, Hu M, Shu W-S. "Microbial communities evolve faster in extreme environments". Scientific Reports, 4(1), 6205, 2014.
29. [29] Güner ED. "Environmental risk assessment for biological wastewater treatment plant". Pamukkale Universitesi Muhendislik Bilimleri Dergisi, 24(3), 476-480, 2018.
30. [30] Newhart KB, Holloway RW, Hering AS, Cath TY. "Datadriven performance analyses of wastewater treatment plants: A review". Water Research, 157, 498-513, 2019.
31. [31] Alisawi HAO. "Performance of wastewater treatment during variable temperature". Applied Water Science, 10(4), 89, 2020.
32. [32] Gude VG. "Energy and water autarky of wastewater treatment and power generation systems". Renewable and Sustainable Energy Reviews, 45, 52-68, 2015.
33. [33] Cui D, Li A, Zhang S, Pang C, Yang J, Guo J, Ma F, Wang J, Ren N. "Microbial community analysis of three municipal wastewater treatment plants in winter and spring using culture-dependent and culture-independent methods". World Journal of Microbiology and Biotechnology, 28(6), 2341-2353, 2012.
34. [34] Ai C, Yan Z, Zhou H, Hou S, Chai L, Qiu G, Zeng W. "Metagenomic insights into the effects of seasonal temperature variation on the activities of activated sludge". Microorganisms, 7(12), 713, 2019.
35. [35] Sobotka D, Zhai J, Makinia J. "Generalized temperature dependence model for anammox process kinetics". Science of the Total Environment, 775, 145760, 2021.
36. [36] O’Flaherty E, Gray NF. "A comparative analysis of the characteristics of a range of real and synthetic wastewaters". Environmental Science and Pollution Research, 20(12), 8813-8830, 2013.
37. [37] Madeira CL, de Araújo JC. "Inhibition of anammox activity by municipal and industrial wastewater pollutants: A review". Science of the Total Environment, 799, 149449, 2021.
38. [38] Can S, Sari T, Akgul D. "Recovery profile of anaerobic ammonium oxidation (anammox) bacteria inhibited by 11 zno nanoparticles". Water Science and Technology, 85(1), 342-353, 2021.
39. [39] Sari T, Akgul D, Mertoglu B. "Long-term response of anammox process to hydrazine under different exposure strategies". Journal of Environmental Chemical Engineering, 12(5), 113600, 2024.
40. [40] Kaewyai J, Noophan P, Wantawin C, Munakata-Marr J. "Recovery of enriched anammox biofilm cultures after storage at cold and room temperatures for 164 days". International Biodeterioration & Biodegradation, 137, 1-7, 2019.
41. [41] Wan K, Yu Y, Hu J, Liu X, Deng X, Yu J, Chi R, Xiao C. "Recovery of anammox process performance after substrate inhibition: Reactor performance, sludge morphology, and microbial community". Bioresource Technology, 357, 127351, 2022.
42. [42] Chen X, Liu L, Bi Y, Meng F, Wang D, Qiu C, Yu J, Wang S. "The recovery of a sequencing biofilm batch reactor— anammox system: Performance, metabolic characteristics, and microbial community analysis". Sustainability, 15(13), 10454, 2023.
43. [43] Mehrani M-J, Sobotka D, Kowal P, Ciesielski S, Makinia J. "The occurrence and role of nitrospira in nitrogen removal systems". Bioresource Technology, 303, 122936, 2020.
44. [44] Braker G, Conrad R. Chapter 2 - diversity, structure, and size of n2o-producing microbial communities in soils— what matters for their functioning? Editors: Laskin AI, Sariaslani S, Gadd GM. Advances in applied microbiology, 33-70, Academic Press, 2011.
45. [45] Cevallos MA, Degli Esposti M. "New alphaproteobacteria thrive in the depths of the ocean with oxygen gradient". Microorganisms, 10(2), 2022.
46. [46] Kersters K, De Vos P, Gillis M, Swings J, Vandamme P, Stackebrandt E. Introduction to the proteobacteria. Editors: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E. The prokaryotes: Volume 5: Proteobacteria: Alpha and beta subclasses, 3-37, New York, Springer New York, 2006.
47. [47] Willis AD. "Rarefaction, alpha diversity, and statistics". Frontiers in Microbiology, 10, 2019.
48. [48] Feranchuk S, Belkova N, Potapova U, Kuzmin D, Belikov S. "Evaluating the use of diversity indices to distinguish between microbial communities with different traits". Research in Microbiology, 169(4), 254-261, 2018.
49. [49] Diserud OH, Odegaard F. "A multiple-site similarity measure". Biology Letters, 3(1), 20-22, 2007.
50. [50] Gülsoy S, Özkan K. "Tür çeşitliliğinin ekolojik açıdan önemi ve kullanılan bazı indisler". Turkish Journal of Forestry, 9(1), 168-178, 2009.
51. [51] Stegenta-Dąbrowska S, Sobieraj K, Rosik J, Sidełko R, Valentin M, Białowiec A. "The development of anammox and chloroflexi bacteria during the composting of sewage sludge". Sustainability, 14(16), 10248, 2022.
52. [52] Jetten MS, Op den Camp HJ, Kuenen JG, Strous M. "“Candidatus brocadiaceae” fam. Nov". Bergey's manual of systematics of Archaea and Bacteria, 1-10, 2015.
53. [53] Kindaichi T, Yuri S, Ozaki N, Ohashi A. "Ecophysiological role and function of uncultured chloroflexi in an anammox reactor". Water Science and Technology, 66(12), 2556-2561, 2012.
54. [54] Bovio-Winkler P, Guerrero LD, Erijman L, Oyarzúa P, Suárez-Ojeda ME, Cabezas A, Etchebehere C. "Genomecentric metagenomic insights into the role of chloroflexi in anammox, activated sludge and methanogenic reactors". BMC Microbiology, 23(1), 45, 2023.
55. [55] Wong LL, Lu Y, Ho JCS, Mugunthan S, Law Y, Conway P, Kjelleberg S, Seviour T. "Surface-layer protein is a public-good matrix exopolymer for microbial community organisation in environmental anammox biofilms". Isme j, 17(6), 803-812, 2023.
56. [56] Spieck E, Spohn M, Wendt K, Bock E, Shively J, Frank J, Indenbirken D, Alawi M, Lücker S, Hüpeden J. "Extremophilic nitrite-oxidizing chloroflexi from yellowstone hot springs". The ISME Journal, 14(2), 364- 379, 2020.
57. [57] Oshiki M, Takaki Y, Hirai M, Nunoura T, Kamigaito A, Okabe S. "Metagenomic analysis of five phylogenetically distant anammox bacterial enrichment cultures". Microbes and Environments, 37(3), 2022.
58. [58] Su Z, Zhao J, Lu Z, Wang M, Guo C, Song X, Guo X, Cai M, Wu Z. "The effects of different temperature conditions on sludge characteristics and microbial communities of nitritation denitrification". Journal of Water Process Engineering, 50, 103283, 2022.
59. [59] Thiel V, Fukushima S-I, Kanno N, Hanada S. Chloroflexi. Editor: Schmidt TM. Encyclopedia of microbiology. 4th ed, 651-662, Oxford, Academic Press, 2019.
60. [60] Zhang Q, Zhang J, Zhao L, Liu W, Chen L, Cai T, Ji X-M. "Microbial dynamics reveal the adaptation strategies of ecological niche in distinct anammox consortia under mainstream conditions". Environmental Research, 215, 114318, 2022.
61. [61] Ya T, Huang Y, Wang K, Wang J, Liu J, Hai R, Zhang T, Wang X. "Functional stability correlates with dynamic microbial networks in anammox process". Bioresource Technology, 370, 128557, 2023.
62. [62] Zhang Q, Zheng J, Zhao L, Liu W, Chen L, Cai T, Ji X-M. "Succession of microbial communities reveals the inevitability of anammox core in the development of anammox processes". Bioresource Technology, 371, 128645, 2023.
63. [63] Pekyavas G, Yangin-Gomec C. "Response of anammox bacteria to elevated nitrogen and organic matter in predigested chicken waste at a long-term operated uasb reactor initially seeded by methanogenic granules". Bioresource Technology Reports, 7, 100222, 2019.
64. [64] Li H, Yang Q, Li J, Gao H, Li P, Zhou H. "The impact of temperature on microbial diversity and aoa activity in the tengchong geothermal field, china". Scientific Reports, 5(1), 17056, 2015.
65. [65] Pitombo LM, do Carmo JB, de Hollander M, Rossetto R, López MV, Cantarella H, Kuramae EE. "Exploring soil microbial 16s rrna sequence data to increase carbon yield and nitrogen efficiency of a bioenergy crop". GCB Bioenergy, 8(5), 867-879, 2016.
66. [66] Hosokawa S, Kuroda K, Narihiro T, Aoi Y, Ozaki N, Ohashi A, Kindaichi T. "Cometabolism of the superphylum patescibacteria with anammox bacteria in a long-term freshwater anammox column reactor". Water, 13(2), 208, 2021.
67. [67] Zhu W, Zhang X, Yuan S, Sang W, Tang X, Zhang S. "Effects of different aeration strategies on removal of organics, nitrogen and phosphorus in sequencing batch biofilm reactor (sbbr): Performance, microbial community and nitrogen cycling pathways". Journal of Water Process Engineering, 49, 103118, 2022.
68. [68] Ya T, Liu J, Zhang M, Wang Y, Huang Y, Hai R, Zhang T, Wang X. "Metagenomic insights into the symbiotic relationship in anammox consortia at reduced temperature". Water Research, 225, 119184, 2022.
69. [69] Zhang H, Du B, Li D, Peng Z, An Z. "Enhancing pollutant removal and desalination in microbial desalination cells using a rotating algal biofilm cathode". Desalination, 586, 117832, 2024.
70. [70] Oba K, Suenaga T, Yasuda S, Kuroiwa M, Hori T, Lackner S, Terada A. "Quest for nitrous oxide-reducing bacteria present in an anammox biofilm fed with nitrous oxide". Microbes Environ, 39(1), 2024.
71. [71] Pimenov NV, Nikolaev YA, Dorofeev AG, Grachev VA, Kallistova AY, Mironov VV, Vanteeva AV, Grigor’eva NV, Berestovskaya YY, Gruzdev EV, Begmatov SA, Ravin NV, Mardanov AV. "Bioaugmentation of anammox activated sludge with a nitrifying bacterial community as a way to increase the nitrogen removal efficiency". Microbiology, 91(2), 133-142, 2022.
72. [72] Peeters SH, van Niftrik L. "Trending topics and open questions in anaerobic ammonium oxidation". Current Opinion in Chemical Biology, 49, 45-52, 2019.
73. [73] Quan ZX, Rhee SK, Zuo JE, Yang Y, Bae JW, Park JR, Lee ST, Park YH. "Diversity of ammonium-oxidizing bacteria in a granular sludge anaerobic ammonium-oxidizing (anammox) reactor". Environmental Microbiology, 10(11), 3130-3139, 2008.
74. [74] Kartal B, Rattray J, van Niftrik LA, van de Vossenberg J, Schmid MC, Webb RI, Schouten S, Fuerst JA, Damsté JS, Jetten MSM, Strous M. "Candidatus “anammoxoglobus propionicus” a new propionate oxidizing species of anaerobic ammonium oxidizing bacteria". Systematic and Applied Microbiology, 30(1), 39-49, 2007.
75. [75] Schmid M, Walsh K, Webb R, Rijpstra WI, van de PasSchoonen K, Verbruggen MJ, Hill T, Moffett B, Fuerst J, Schouten S, Sinninghe Damsté JS, Harris J, Shaw P, Jetten M, Strous M. "Candidatus “scalindua brodae”, sp. Nov., candidatus “scalindua wagneri”, sp. Nov., two new species of anaerobic ammonium oxidizing bacteria". Systematic and Applied Microbiology, 26(4), 529-538, 2003.
76. [76] Schmid M, Twachtmann U, Klein M, Strous M, Juretschko S, Jetten M, Metzger JW, Schleifer K-H, Wagner M. "Molecular evidence for genus level diversity of bacteria capable of catalyzing anaerobic ammonium oxidation". Systematic and Applied Microbiology, 23(1), 93-106, 2000.
77. [77] Kartal B, van Niftrik L, Rattray J, van de Vossenberg JLCM, Schmid MC, Sinninghe Damsté J, Jetten MSM, Strous M. "Candidatus ‘brocadia fulgida’: An autofluorescent anaerobic ammonium oxidizing bacterium". FEMS Microbiology Ecology, 63(1), 46-55, 2008.
78. [78] Yang Y, Lu Z, Azari M, Kartal B, Du H, Cai M, Herbold CW, Ding X, Denecke M, Li X, Li M, Gu J-D. "Discovery of a new genus of anaerobic ammonium oxidizing bacteria with a mechanism for oxygen tolerance". Water Research, 226, 119165, 2022.
79. [79] Oshiki M, Satoh H, Okabe S. "Ecology and physiology of anaerobic ammonium oxidizing bacteria". Environmental Microbiology, 18(9), 2784-2796, 2016.
80. [80] Fu J-J, Wang Y, Yang J-H, Huang D-Q, Zhang Q, Huang Y, Chen J-R, Fan N-S, Jin R-C. "Mitigating the detrimental effects of salt stress on anammox process: A comparison between glycine betaine and mannitol". Science of the Total Environment, 851, 158221, 2022.
81. [81] Xu L-Z-J, Zhang Q, Fu J-J, Zhang J-T, Zhao Y-H, Jin L-Y, Fan N-S, Huang B-C, Jin R-C. "Deciphering the microbial community and functional genes response of anammox sludge to sulfide stress". Bioresource Technology, 302, 122885, 2020.
82. [82] Wang Y, Wang X, Niu J. "Implemented impediment of extracellular electron transfer–dependent anammox process :Unstable nitrogen removal efficiency and decreased abundance of anammox bacteria". Chemosphere, 337, 139415, 2023.
83. [83] Tian S, Tian Z, Yang H, Yang M, Zhang Y. "Detection of viable bacteria during sludge ozonation by the combination of atp assay with pma-miseq sequencing". Water, 9(3), 2017.
84. [84] Yang R, Mao W, Wang X, Zhang Z, Wu J, Chen S. "Response and adaptation of microbial community in a canon reactor exposed to an extreme alkaline shock". Archaea, 2020, 8888615, 2020.
85. [85] Sari T, Akgul D, Mertoglu B. "Enhancement of hydrazine accumulation in anammox bioreactors". Chemosphere, 359, 142293, 2024.
86. [86] Ya T, Du S, Li Z, Liu S, Zhu M, Liu X, Jing Z, Hai R, Wang X. "Successional dynamics of molecular ecological network of anammox microbial communities under elevated salinity". Water Research, 188, 116540, 2021.
87. [87] Allegue T, Arias A, Fernandez-Gonzalez N, Omil F, Garrido JM. "Enrichment of nitrite-dependent anaerobic methane oxidizing bacteria in a membrane bioreactor". Chemical Engineering Journal, 347, 721-730, 2018.
88. [88] Hu L, Cheng X, Qi G, Zheng M, Dang Y, Li J, Xu K. "Achieving ammonium removal through anammoxderived feammox with low demand of fe(iii)". Frontiers in Microbiology, 13, 918634, 2022.
89. [89] Yang N, Zhan G, Li D, He X, Zhang Y, Jiang Q, Liu H, Wang C. "Performance and microbial community of a novel non-aeration-based up-flow bioelectrochemical filter (ubef) treating real domestic wastewater". Chemical Engineering Journal, 348, 271-280, 2018.
90. [90] Zhang S, Zhang Z, Xia S, Ding N, Liao X, Yang R, Chen M, Chen S. "The potential contributions to organic carbon utilization in a stable acetate-fed anammox process under low nitrogen-loading rates". Science of the Total Environment, 784, 147150, 2021.
91. [91] Ahmad HA, Guo B, Zhuang X, Zhao Y, Ahmad S, Lee T, Zhu J, Dong Y, Ni S-Q. "A twilight for the complete nitrogen removal via synergistic partial-denitrification, anammox, and dnra process". npj Clean Water, 4(1), 31, 2021.
92. [92] Ribeiro H, Wijaya IMW, Soares-Santos V, Soedjono ES, Slamet A, Teixeira C, Bordalo AA. "Microbial community composition, dynamics, and biogeochemistry during the start-up of a partial nitritation-anammox pathway in an upflow reactor". Sustainable Environment Research, 32(1), 18, 2022.
93. [93] Zhang J-F, Lai C-Y, Cao X-X, Hartmann EM, Zhao H-P. "High ammonia loading rate and biofilm reattachment initiated partial nitrification and anammox in a membrane aerated biofilm reactor". Journal of Water Process Engineering, 58, 104829, 2024.
94. [94] Ding J, Qin F, Li C, Tang M. "Long-term effect of acetate and biochar addition on enrichment and activity of denitrifying anaerobic methane oxidation microbes". Chemosphere, 338, 139642, 2023.
95. [95] Sun L, Toyonaga M, Ohashi A, Matsuura N, Tourlousse DM, Meng XY, Tamaki H, Hanada S, Cruz R, Yamaguchi T, Sekiguchi Y. "Isolation and characterization of flexilinea 13 flocculi gen. Nov., sp. Nov., a filamentous, anaerobic bacterium belonging to the class anaerolineae in the phylum chloroflexi". International Journal of Systematic and Evolutionary Microbiology, 66(2), 988-996, 2016.
96. [96] Lawson CE, Wu S, Bhattacharjee AS, Hamilton JJ, McMahon KD, Goel R, Noguera DR. "Metabolic network analysis reveals microbial community interactions in anammox granules". Nature Communications, 8(1), 15416, 2017.
97. [97] Speth DR, in ’t Zandt MH, Guerrero-Cruz S, Dutilh BE, Jetten MSM. "Genome-based microbial ecology of anammox granules in a full-scale wastewater treatment system". Nature Communications, 7(1), 11172, 2016.
98. [98] Qiu J, Li T, Lü F, Huang Y, Li C, Zhang H, Shao L, He P. "Molecular behavior and interactions with microbes during anaerobic degradation of bio-derived dom in waste leachate". Journal of Environmental Sciences, 126, 174- 183, 2023.
99. [99] Cheng L, Yang W, Liang H, Nabi M, Li Y, Wang H, Hu J, Chen T, Gao D. "Nitrogen removal from mature landfill leachate through enhanced partial nitrification-anammox process in an innovative multi-stage fixed biofilm reactor". Science of the Total Environment, 877, 162959, 2023.
100. [100] Luo Y, Yi K, Zhang X, Li B, Cao R, Pang Y, Li M, Hou C, Lv J, Li X, Li D. "Simultaneous partial nitrification, denitrification, and phosphorus removal in sequencing batch reactors via controlled reduced aeration and shortterm sludge retention time decrease". Journal of Environmental Management, 344, 118598, 2023.
101. [101] Li J, Chen X, Yang Z, Liu Z, Chen Y, Wang Y-e, Xie H. "Denitrification performance and mechanism of sequencing batch reactor with a novel iron-polyurethane foam composite carrier". Biochemical Engineering Journal, 176, 108209, 2021.
102. [102] Li S, Li J, Yang S, Zhang Q, Li X, Zhang L, Peng Y. "Rapid achieving partial nitrification in domestic wastewater: Controlling aeration time to selectively enrich ammonium oxidizing bacteria (aob) after simultaneously eliminating aob and nitrite oxidizing bacteria (nob)". Bioresource Technology, 328, 124810, 2021.
103. [103] Li Y, Wang J, Hua M, Yao X, Zhao Y, Hu J, Xi C, Hu B. "Strategy for denitrifying anaerobic methane-oxidizing bacteria growing under the oxygen-present condition". Science of the Total Environment, 742, 140476, 2020.
104. [104] Toumi M, Abbaszade G, Sbaoui Y, Farkas R, Ács É, Jurecska L, Tóth E. "Cultivation and molecular studies to reveal the microbial communities of groundwaters discharge located in hungary". Water, 13(11), 1533, 2021.
105. [105] Wang C, Yu G, Yang F, Wang J. "Formation of anaerobic granules and microbial community structure analysis in anaerobic hydrolysis denitrification reactor". Science of the Total Environment, 737, 139734, 2020.