长期运行厌氧氨氧化系统沿程不同形态污泥微生物群落结构及功能解析

闫冰; 聂含冰; 付嘉琦; 韩佩; 易其臻; 桂双林; 夏嵩

PDF(3821 KB)

中国环境科学 ›› 2025, Vol. 45 ›› Issue (2) : 1052-1062.

环境微生物

长期运行厌氧氨氧化系统沿程不同形态污泥微生物群落结构及功能解析

闫冰^1,2,3, 聂含冰^1,2,3, 付嘉琦^1,2,3, 韩佩^1,3, 易其臻^1,3, 桂双林^1,2,3, 夏嵩^1,2,3

作者信息 +

Deciphering the microbial community structure and function of sludge with different morphologies along the ANAMMOX reactor under long-term operation

YAN Bing^1,2,3, NIE Han-bing^1,2,3, FU Jia-qi^1,2,3, HAN Pei^1,3, YI Qi-zhen^1,3, GUI Shuang-lin^1,2,3, XIA Song^1,2,3

Author information +

文章历史 +

摘要

为深入理解厌氧氨氧化系统沿程微生物群落特征,考察了长期运行的升流式厌氧污泥床(UASB)厌氧氨氧化系统沿程方向上不同形态污泥微生物群落多样性、结构、物种网络及功能特征.结果表明,厌氧氨氧化系统沿程方向上不同形态污泥AnAOB基因丰度存在显著差异(P<0.05),底部颗粒污泥(KL)中AnAOB基因丰度为2.12x10¹⁰copies/g VSS,显著高于系统沿程其它形态污泥(P<0.05);沿程方向上不同形态污泥微生物多样性存在显著差异(P<0.05),但未呈现出明显的变化规律.不同形态污泥优势菌门(相对丰度>1%)为绿弯菌门(Chloroflexi)、浮霉菌门(Planctomycetota)、变形菌门(Proteobacteria)、拟杆菌门(Bacteroidota)、酸杆菌门(Acidobacteriota)和放线菌门(Actinobacteriota),相对丰度排名前10的菌属为norank_f__norank_o__SBR1031、Candidatus_Kuenenia、norank_f__Anaerolineaceae、Nitrosomonas、Limnobacter、norank_f__PHOS-HE36、Denitratisoma、Denitratisoma和OLB13norank_f__A4b,相对丰度在不同样品间存在显著差异(P<0.05);沿程方向不同形态污泥微生物群落结构间存在显著差异(P=0.001).网络分析发现,不同形态污泥微生物网络结构和拓扑性质存在差异,其中沉淀池内絮状泥(CD)、出水管内生物膜(XK)的平均度和中心紧密度高于沿程其他样品,微生物群落间具有更紧密的关联性.PICRUSt2分析发现,与代谢相关的通路功能基因丰度明显大于其他功能基因丰度,并且沿程不同形态污泥间代谢功能基因丰度存在显著差异(P<0.05),氮素代谢功能基因hao相对丰度远高于norB,同时narG基因丰度是nirS丰度的20.8~733.9倍,系统内部分亚硝化和部分反硝化功能活跃.厌氧氨氧化系统沿程方向上不同形态污泥微生物群落存在明显的异质性特征,但未呈现出明显的规律性.

Abstract

To understand the characteristics of microbial communities along the anaerobic ammonia oxidation (ANAMMOX) system, this study explored the diversity, structure, species network, and functional features of microbial communities in sludge with different morphologies along the flow direction in a long-term operated up-flow anaerobic sludge bed (UASB) ANAMMOX system. The results showed significant differences (P<0.05) in the abundance of AnAOB genes in sludge with different morphologies along the anaerobic ammonia oxidation system. The abundance of AnAOB genes in the bottom granular sludge (KL) was 2.12×10¹⁰copies/g VSS, which was significantly higher (P<0.05) than that in sludge with other morphologies along the system. There were significant differences (P<0.05) in the microbial diversity of sludge with different morphologies along the system, but no obvious change patterns were observed. The dominant bacterial phyla (relative abundance>1%) in sludge with different morphologies were Chloroflexi, Planctomycota, Proteobacteria, Bacteroidota, Acidobacteriota, and Actinobacteriota. The top 10genera in terms of relative abundance were norank_f_norank_o_SBR1031, Candidatus_Kuenenia, norank_f_Anaerolineaceae, Nitrosomonas, Limnobacteriota, norank_f_PHOS-HE36, Denitratisoma, Denitratisoma, and OLB13norank_f_ A4b. There were significant differences (P<0.05) in relative abundance among different samples. There were significant differences (P=0.001) in the microbial community structure of sludge with different morphology along the process. Network analysis found differences in the structure and topological properties of microbial networks among different morphologies of sludge. Specifically, the average degree and center tightness of flocculent sludge in sedimentation tank (CD) and biofilm in the effluent pipe (XK) were higher than those in other samples along the system, indicating a closer correlation among microbial communities. PICRUSt2analysis revealed that the abundance of functional genes related to metabolic pathways was significantly higher than that of other functional genes. Additionally, there were significant differences (P<0.05) in metabolic functional genes abundance among different morphologies of sludge along the process. The relative abundance of nitrogen metabolism functional gene hao was much higher than that of norB, while the abundance of the narG gene was 20.8 to 733.9times that of nirS, indicating activeness of partial nitritation and partial denitrification functions within the system. The anaerobic ammonia oxidation system exhibits significant heterogeneity in the microbial communities of sludge with different morphologies along the process, but no distinct regularity is observed.

导出引用

闫冰, 聂含冰, 付嘉琦, 韩佩, 易其臻, 桂双林, 夏嵩. 长期运行厌氧氨氧化系统沿程不同形态污泥微生物群落结构及功能解析[J]. 中国环境科学. 2025, 45(2): 1052-1062

YAN Bing, NIE Han-bing, FU Jia-qi, HAN Pei, YI Qi-zhen, GUI Shuang-lin, XIA Song. Deciphering the microbial community structure and function of sludge with different morphologies along the ANAMMOX reactor under long-term operation[J]. China Environmental Science. 2025, 45(2): 1052-1062

中图分类号： X172

参考文献

[1] Wu P, Chen J J, Garlapati V K, et al. Novel insights into Anammox-based processes: A critical review [J]. Chemical Engineering Journal, 2022,444:136534.
[2] Lackner S, Gilbert E M, Vlaeminck S E, et al. Full-scale partial nitritation/anammox experiences-An application survey [J]. Water Research, 2014,55:292-303.
[3] Tomaszewski M, Cema G, Ziembińska-Buczyńska A. Influence of temperature and pH on the anammox process: A review and meta-analysis [J]. Chemosphere, 2017,182:203-214.
[4] 李泽兵,刘常敬,赵白航,等.多基质时厌氧氨氧化菌、异养反硝化污泥活性及抑制特征 [J]. 中国环境科学, 2013,33(4):648-654.Li Z B, Liu C J, Zhao B H, et al. Activity and inhibition characteristics of anammox and heterotrophic denitrifier bacteria in a multi-substrate system [J]. China Environmental Science, 2013,33(4):648-654.
[5] 姜滢,郭萌蕾,谢军祥,等.不同培养条件厌氧氨氧化颗粒污泥性质及微生物群落结构差异 [J]. 环境科学, 2020,41(5):2358-2366.Jiang Y, Guo M L, Xie J X, et al. Characteristics of ANAMMOX granular sludge and differences in microbial community structure under different culture conditions [J]. Environmental Science, 2020, 41(5):2358-2368.
[6] 王晗,李瀚翔,陈猷鹏,等.盐度条件下ANAMMOX-EGSB反应器颗粒污泥微生物群落 [J]. 环境科学, 2019,40(4):1906-1913.Wang H, Li H X, Chen Y P, et al. Microbial community of granular sludge in an ANAMMOX-EGSB reactor under saline conditions [J]. Environmental Science, 2019,40(4):1906-1913.
[7] 闫冰,夏嵩,桂双林,等.厌氧氨氧化菌富集培养过程微生物群落结构及多样性 [J]. 环境科学, 2020,41(12):5535-5543.Yan B, Xia S, Gui S L, et al. Microbial community structure and diversity during the enrichment of anaerobic ammonium oxidation bacteria [J]. Environmental Science, 2020,41(12):5535-5543.
[8] 李亚男,姚丽,隋倩雯,等.厌氧氨氧化细菌富集培养过程微生物结构与功能解析 [J]. 环境科学学报, 2021,41(1):92-101.Li Y N, Yao L, Sui Q W, et al. Evolution of microbial structures and functions during the enrichment of anammox bacteria [J] Acta Scientiae Circumstantiae, 2021,41(1):92-101.
[9] 马雨晴,王博,李笑迪.温度与负荷对Anammox菌自富集启动的PDA的影响 [J]. 中国环境科学, 2024,44(3):1307-1313.Ma Y Q, Wang B, Li X D. Effect of temperature and load variation of pda system initiated by anammox bacteria self-enrichment [J]. China Environmental Science, 2024,44(3):1307-1313.
[10] 赵星程,贾方旭,刘晨雨,等.实际规模PN/A工艺载体挂膜性能与微生物群落 [J]. 化工进展, 2024.https://doi.org/10.16085/j.issn.1000-6613.2023-1310.Zhao X C, Jia F X, Liu C Y, et al. Biofilm immobilization effects and microbial communities of the carriers in real scale PN/A process [J]. Chemical Industry and Engineering Progress, 2024.https://doi.org/ 10.16085/j.issn.1000-6613.2023-1310.
[11] Ma Y, Wang B, Li X, et al. Enrichment of anammox biomass during mainstream wastewater treatment driven by achievement of partial denitrification through the addition of bio-carriers [J]. Journal of Environmental Sciences, 2024,137:181-194.
[12] Zhou F, Xiao W Z, Zhou K Y, et al. Performance characteristics and community analysis of a single-stage partial nitritation, anammox and denitratation (SPANADA) integrated process for treating low C/N ratio wastewater [J]. Chemical Engineering Journal, 2022,433(P1): 134452.
[13] Depeng W, Kailong H, Xiwei H, et al. Varied interspecies interactions between anammox and denitrifying bacteria enhanced nitrogen removal in a single-stage simultaneous anammox and denitrification system [J]. Science of the Total Environment, 2021,813:152519.
[14] 常尧枫,郭萌蕾,谢军祥,等.厌氧氨氧化脱氮除碳功能菌群结构及代谢途径 [J]. 中国环境科学, 2022,42(3):1138-1145.Chang Y F, Guo M L, Xie J X, et al. The structure and metabolic pathway of functional bacteria for nitrogen and carbon removal in Anammox [J]. China Environmental Science, 2022,42(3):1138-1145.
[15] 鲍林林,陈婉秋,申今生,等.折流板生物膜反应器快速启动厌氧氨氧化 [J]. 哈尔滨工业大学学报, 2016,48(2):114-118.Bao L L, Chen W Q, Shen J S, et al. Study on the fast startup of the anammox process by baffled biofilm reactor [J]. Journal of Harbin Institute of Technology, 2016,48(2):114-118.
[16] 秦榕,宋佳宇,齐碧薇,等.厌氧氨氧化反应器菌群动态演替分析 [J]. 环境科学与技术, 2020,43(S1):23-28.Qin R, Song J Y, Qi B W, et al. Dynamic succession analysis of bacteria in anaerobic ammonia oxidation reactor for refining wastewater treatment [J]. Environmental Science and Technology, 2020,43(S1):23-28.
[17] 闫冰,付嘉琦,夏嵩,等.厌氧氨氧化启动过程细菌群落多样性及PICRUSt2功能预测分析 [J]. 环境科学, 2021,42(8):3875-3885.Yan B, Fu J Q, Xia S, et al. Diversity and PICRUSt2-based predicted functional analysis of bacterial communities during the start-up of ANAMMOX [J]. Environmental Science, 2021,42(8):3875-3885.
[18] Van D G A A, De Bruijn P, Robertson L A, et al. Autotrophic growth of anaerobic ammonium-oxidizing micro-organisms in a fluidized bed reactor [J]. Microbiology, 1996,142(8):2187-2196.
[19] Caporaso J G, Lauber C L, Walters W A, et al. Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample [J]. Proceedings of the National Academy of Sciences of the United States of America, 2011,108(S11):4516-4522.
[20] Schmid M C, Maas B, Dapena A, et al. Biomarkers for in situ detection of anaerobic ammonium-oxidizing (Anammox) bacteria [J]. Applied and Environmental Microbiology, 2005,71(4):1677-1684.
[21] Bolyen E, Rideout J R, Dillon M R, et al. Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2 [J]. Nature Biotechnology, 2019,37(8):852-857.
[22] Callahan B J, McMurdie P J, Rosen M J, et al. DADA2: High-resolution sample inference from Illumina amplicon data [J]. Nature Methods, 2016,13(7):581-583.
[23] Katoh K, Misawa K, Kuma K, et al. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform [J]. Nucleic Acids Research, 2002,30(14):3059-3066.
[24] Price M N, Dehal P S, Arkin A P. FastTree 2--Approximately maximum-likelihood trees for large alignments [J]. Plos One, 2010, 5(3):e9490.
[25] 梁子阳,彭嘉敏,张泳纯,等.低进水浓度CASS工艺沿程污染物去除特征及微生物群落变化 [J]. 环境工程学报, 2023,17(10):3219-3230.Liang Z Y, Peng J M, Zhang Y C, et al. Pollutant removal characteristics and microbial community changes along the CASS process with low concentration influent [J]. Chinese Journal of Environmental Engineering, 2023,17(10):3219-3230.
[26] Feng Y, Zhao Y, Guo Y, et al. Microbial transcript and metabolome analysis uncover discrepant metabolic pathways in autotrophic and mixotrophic anammox consortia [J]. Water Research, 2018,128:402-411.
[27] Lawson C, Wu S, Bhattacharjee A S, et al. Metabolic network analysis reveals microbial community interactions in anammox granules [J]. Nature Communications, 2017,8(1):15416.
[28] 曹雁,王桐屿,秦玉洁,等.厌氧氨氧化反应器脱氮性能及细菌群落多样性分析 [J]. 环境科学, 2017,38(4):1544-1550.Cao Y, Wang T Y, Qin Y J, et al. Nitrogen removal characteristics and diversity of microbial community in ANAMMOX reactor [J]. Environmental Science, 2017,38(4):1544-1550.
[29] Kindaichi T, Yuri S, Ozaki N, et al. Ecophysiological role and function of uncultured Chloroflexi in an anammox reactor [J]. Water Science and Technology, 2012,66(12):2556-2561.
[30] Björnsson L, Hugenholtz P, Tyson G W, et al. Filamentous Chloroflexi (green non-sulfur bacteria) are abundant in wastewater treatment processes with biological nutrient removal [J]. Microbiology, 2002, 148(8):2309-2318.
[31] Zeng T, Li D, Liao W, et al. Nitrogen removal and functional bacteria distribution of ANAMMOX at ambient temperature [J]. Journal of Water Reuse and Desalination, 2016,6(4):476-483.
[32] Lu H, Chandran K, Stensel D. Microbial ecology of denitrification in biological wastewater treatment [J]. Water Research, 2014,64:237-254.
[33] Cas S B, Du R, Baikun L, et al. High-throughput profiling of microbial community structures in an ANAMMOX-UASB reactor treating high-strength wastewater [J]. Applied Microbiology and Biotechnology, 2016,100(14):6457-6467.
[34] Zhang L, Narita Y, Gao L, et al. Microbial competition among anammox bacteria in nitrite-limited bioreactors [J]. Water Research, 2017,125:249-258.
[35] Wang C, Liu S T, Xu X C, et al. Achieving mainstream nitrogen removal through simultaneous partial nitrification, anammox and denitrification process in an integrated fixed film activated sludge reactor [J]. Chemosphere, 2018,203:457-466.
[36] Xia Y, Wang Y, Wang Y, et al. Cellular adhesiveness and cellulolytic capacity in Anaerolineae revealed by omics-based genome interpretation [J]. Biotechnology for Biofuels, 2016,9(1):111.
[37] Xiang T, Gao D. Comparing two hydrazine addition strategies to stabilize mainstream deammonification: Performance and microbial community analysis[J]. Bioresource Technology, 2019,289:121710.
[38] Ma B, Wang H Z, Dsouza M, et al. Geographic patterns of co-occurrence network topological features for soil microbiota at continental scale in eastern China [J]. The ISME Journal, 2016,10(8): 1891-1901.
[39] Liu Y, Sun J, Peng L, et al. Assessment of heterotrophic growth supported by soluble microbial products in anammox biofilm using multidimensional modeling [J]. Scientific Reports, 2016,6(1):27576.
[40] Zhao Y, Liu S, Jiang B, et al. Genome-centered metagenomics analysis reveals the symbiotic organisms possessing ability to cross-feed with anammox bacteria in anammox consortia [J]. Environmental Science and Technology, 2018,52(19):11285-11296.