Abstract:A series of batch experiments were conducted to investigate the responses of anaerobic digestion (AD) microbial community exposed to ammonia (TAN) stress. In combination with Metagenomic sequencing analysis, this paper investigated the responses of process parameters, community structure/composition and functional gene/metabolic pathways of AD to the process disturbance induced by different TAN concentrations. The overall performance of AD system was gradually optimized at the concentration of 3000mg/L TAN while when TAN reached 6000mg/L, an accumulation of propionate and butyrate was observed with a concomitant decrease of methane yield. At both concentration levels, the resistance and redundancy of hydrolytic and acidogenic bacteria can ensure their metabolic performance for substrate degradation. However, the total relative abundance of acetogens and methanogens at the low concentration level continued to increase, whereas a low increase rate and even a decrease in the total relative abundance was observed at the high level, suggesting the potential process inhibition. By further analyzing the functional gene changes in the two process stages, The methanogenesis and syntrophic acetate oxidation pathways showed resistance and redundancy under the ammonia stress, but the high concentration of TAN (6000mg/L) inhibited the expression of key genes ACADS and PCCA in the syntrophic degradation of butyrate and propionate. Therefore, the key contributor to the ammonia inhibition turned out to be acetogenesis.
彭韵, 李蕾, 伍迪, 杨屏锦, 彭绪亚, 王小铭. 微生物群落对氨胁迫响应的宏基因组学研究[J]. 中国环境科学, 2022, 42(2): 777-786.
PENG Yun, LI Lei, WU Di, YANG Ping-jin, PENG Xu-ya, WANG Xiao-ming. Metagenomic analysis on the responses of microbial community to ammonia stress. CHINA ENVIRONMENTAL SCIENCECE, 2022, 42(2): 777-786.
Wu D, Li L, Zhao X F, et al. Anaerobic digestion:A review on process monitoring[J]. Renewable and Sustainable Energy Reviews, 2019,103:1-12.
[2]
Li L, Peng X Y, Wang X M, et al. Anaerobic digestion of food waste:A review focusing on process stability[J]. Bioresource Technology, 2018,248:20-28.
[3]
Chen H, Wang W, Xue L N, et al. Effects of ammonia on anaerobic digestion of food waste:Process performance and microbial community[J]. Energy & Fuels, 2016,30(7):5749-5757.
[4]
Bi S J, Mw D, Westerholm M, et al. The metabolic performance and microbial communities of anaerobic digestion of chicken manure under stressed ammonia condition:A case study of a 10-year successful biogas plant[J]. Renewable Energy, 2021,167:644-651.
[5]
Wijesinghe D T N, Suter H C, Scales P J, et al. Lignite addition during anaerobic digestion of ammonium rich swine manure enhances biogas production[J]. Journal of Environmental Chemical Engineering, 2021, 9(1):104669.
[6]
Meng X S, Sui Q W, Liu J B, et al. Relieving ammonia inhibition by zero-valent iron (ZVI) dosing to enhance methanogenesis in the high solid anaerobic digestion of swine manure[J]. Waste Management, 2020,118:452-462.
[7]
Niu Q G, Qiao W, Qiang H, et al. Microbial community shifts and biogas conversion computation during steady, inhibited and recovered stages of thermophilic methane fermentation on chicken manure with a wide variation of ammonia[J]. Bioresource Technology, 2013,146:223-233.
[8]
Yang Z Y, Wang W, He Y F, et al. Effect of ammonia on methane production, methanogenesis pathway, microbial community and reactor performance under mesophilic and thermophilic conditions[J]. Renewable Energy, 2018,125:915-925.
[9]
Bi S J, Qiao W, Xiong L P, et al. Improved high solid anaerobic digestion of chicken manure by moderate in situ ammonia stripping and its relation to metabolic pathway[J]. Renewable Energy, 2020, 146:2380-2389.
[10]
Buhlmann C H, Mickan B S, Jenkins S N, et al. Ammonia stress on a resilient mesophilic anaerobic inoculum:Methane production, microbial community, and putative metabolic pathways[J]. Bioresource Technology, 2019,275:70-77.
[11]
Zhang N, Peng H J, Li Y, et al. Ammonia determines transcriptional profile of microorganisms in anaerobic digestion[J]. Brazilian Journal of Microbiology, 2018,49(4):770-776.
[12]
Peng X Y, Zhang S Y, Li L, et al. Long-term high-solids anaerobic digestion of food waste:Effects of ammonia on process performance and microbial community[J]. Bioresource Technology, 2018,262:148-158.
[13]
Fotidis I A, Karakashev D, Angelidaki I. Bioaugmentation with an acetate-oxidising consortium as a tool to tackle ammonia inhibition of anaerobic digestion[J]. Bioresource Technology, 2013,146:57-62.
[14]
Li Y, Zhang Y, Kong X Y, et al. Effects of ammonia on propionate degradation and microbial community in digesters using propionate as a sole carbon source[J]. Journal of Chemical Technology and Biotechnology, 2017,92(10):2538-2545.
[15]
Bonk F, Popp D, Weinrich S, et al. Ammonia inhibition of anaerobic volatile fatty acid degrading microbial communities[J]. Frontiers in Microbiology, 2018,9:2921.
[16]
Wang H Z, Yan Y C, Gou M, et al. Response of propionate-degrading methanogenic microbial communities to inhibitory conditions[J]. Applied Biochemistry and Biotechnology, 2019,189(1):233-248.
[17]
Zhang H, Peng Y, Yang P J, et al. Response of process performance and microbial community to ammonia stress in series batch experiments[J]. Bioresource Technology, 2020,314:123768.
[18]
Ruiz-Sánchez J, Campanaro S, Guivernau M, et al. Effect of ammonia on the active microbiome and metagenome from stable full-scale digesters[J]. Bioresource Technology, 2018,250:513-522.
[19]
何琴,李蕾,瞿莉,等.餐厨垃圾干式厌氧消化污泥膨胀微生态特征[J]. 中国环境科学. 2018,38(3):1010-1017. He Q, Li L, Qu L, et al. Microbial characteristics of bulking sludge in high-solids anaerobic digestion of kitchen waste[J]. Chinese Environmental Science, 2018,38(3):1010-1017.
[20]
国家环境保护总局水与废水监测分析方法编委会.水与废水监测分析方法[M]. 北京:中国环境科学出版社, 2002. State Environmental Protection Administration. Determination methods for examination of water and wastewater[M]. Beijing:China Environmental Science Press, 2002.
[21]
Chen S S, He J, Wang H Y, et al. Microbial responses and metabolic pathways reveal the recovery mechanism of an anaerobic digestion system subjected to progressive inhibition by ammonia[J]. Chemical Engineering Journal, 2018,350:312-323.
[22]
Ma X X, Yu M, Song N, et al. Effect of ethanol pre-fermentation on organic load rate and stability of semi-continuous anaerobic digestion of food waste[J]. Bioresource Technology, 2020,299:122587.
[23]
Li D, Liu S C, Mi L, et al. Effects of feedstock ratio and organic loading rate on the anaerobic mesophilic co-digestion of rice straw and cow manure[J]. Bioresource Technology, 2015,189:319-326.
[24]
Gomes C S, Strangfeld M, Meyer M. Diauxie studies in biogas production from gelatin and adaptation of the modified Gompertz model:Two-phase Gompertz model[J]. Applied Sciences-basel, 2021, 11(3):1067.
[25]
Gu J, Liu R, Cheng Y, et al. Anaerobic co-digestion of food waste and sewage sludge under mesophilic and thermophilic conditions:Focusing on synergistic effects on methane production[J]. Bioresource Technology, 2020,301:122765.
[26]
Li L, He Q, Zhao X F, et al. Anaerobic digestion of food waste:Correlation of kinetic parameters with operational conditions and process performance[J]. Biochemical Engineering Journal, 2018,130:1-9.
[27]
Yu D W, Zhang J Y, Chulu B, et al. Ammonia stress decreased biomarker genes of acetoclastic methanogenesis and second peak of production rates during anaerobic digestion of swine manure[J]. Bioresource Technology, 2020,317:124012.
[28]
Kurade M B, Saha S, Kim J R, et al. Microbial community acclimatization for enhancement in the methane productivity of anaerobic co-digestion of fats, oil, and grease[J]. Bioresource Technology, 2020,296:122294.
[29]
Tian H L, Fotidis I A, Mancini E, et al. Acclimation to extremely high ammonia levels in continuous biomethanation process and the associated microbial community dynamics[J]. Bioresource Technology, 2018,247:616-623.
[30]
Pan X F, Zhao L, Li C X, et al. Deep insights into the network of acetate metabolism in anaerobic digestion:focusing on syntrophic acetate oxidation and homoacetogenesis[J]. Water Research, 2021, 190:116774.
[31]
Li Y, Sun Y M, Yang G X, et al. Vertical distribution of microbial community and metabolic pathway in a methanogenic propionate degradation bioreactor[J]. Bioresource Technology, 2017,245:1022-1029.
[32]
邓玉营.共培养菌群强化秸秆厌氧消化及微生物学机制研究[D]. 无锡:江南大学, 2017. Deng Y Y. Enhanced anaerobic digestion of straw via co-cultivated consortia and its underlying microbial mechanisms[D]. Wuxi:Jiangnan University, 2017.
[33]
张虹,李蕾,彭韵,等.氨氮对餐厨垃圾厌氧消化性能及微生物群落的影响[J]. 中国环境科学, 2020,40(8):3465-3474. Zhang H, Li L, Peng Y, et al. Effects of ammonia on anaerobic digestion of food waste:Process performance and microbial community[J]. China Environmental Science, 2020,40(8):3465-3474.
[34]
Yin D M, Westerholm M, Qiao W, et al. An explanation of the methanogenic pathway for methane production in anaerobic digestion of nitrogen-rich materials under mesophilic and thermophilic conditions[J]. Bioresource Technology, 2018,264:42-50.
[35]
Du J, Yin Q D, Gu M Q, et al. New insights into the effect of ethanol and volatile fatty acids proportions on methanogenic activities and pathways[J]. Environmental Research, 2021,194:110644.
[36]
Saha S, Jeon B H, Kurade M B, et al. Interspecies microbial nexus facilitated methanation of polysaccharidic wastes[J]. Bioresource Technology, 2019,289:121638.
[37]
Xia X X, Zhang J C, Song T Z, et al. Stimulation of Smithella -dominating propionate oxidation in a sediment enrichment by magnetite and carbon nanotubes:Propionate syntrophy in the presence of conductive materials[J]. Environmental Microbiology Reports, 2019,11(2):236-248.
[38]
孙航宇,杨紫怡,李潇男,等. ADM1模型对生物强化厌氧产甲烷体系的模拟[J]. 中国环境科学, 2020,40(3):1049-1058. Sun H Y, Yang Z Y, Li X N, et al. Simulation of anaerobic digestion based on bioaugmentation by ADM1[J]. Chinese Environmental Science, 2020,40(3):1049-1058.
[39]
Westerholm M, Dolfing J, Schnurer A. Growth characteristics and thermodynamics of syntrophic acetate oxidizers[J]. Environmental Science & Technology, 2019,53(9):5512-5520.
[40]
Westerholm M, Moestedt J, Schnurer A. Biogas production through syntrophic acetate oxidation and deliberate operating strategies for improved digester performance[J]. Applied Energy, 2016,179:124-135.
[41]
Ruiz-Sánchez J, Guivernau M, Fernandez B, et al. Functional biodiversity and plasticity of methanogenic biomass from a full-scale mesophilic anaerobic digester treating nitrogen-rich agricultural wastes[J]. Science of the Total Environment, 2019,649:760-769.
[42]
方晓瑜,李家宝,芮俊鹏,等.产甲烷生化代谢途径研究进展[J]. 应用与环境生物学报, 2015,21(1):1-9. Fang X Y, Li J B, Rui J P, et al. Research progress in biochemical pathways of methanogenesis[J]. Chinese Journal of Applied and Environmental Biology, 2015,21(1):1-9.
[43]
Yan M, Treu L, Campanaro S, et al. Effect of ammonia on anaerobic digestion of municipal solid waste:inhibitory performance, bioaugmentation and microbiome functional reconstruction[J]. Chemical Engineering Journal, 2020,401:126159.
[44]
Leng L, Yang P X, Singh S, et al. A review on the bioenergetics of anaerobic microbial metabolism close to the thermodynamic limits and its implications for digestion applications[J]. Bioresource Technology, 2018,247:1095-1106.