微量元素对蔬菜废弃物连续厌氧消化系统微生物群落结构的影响
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摘要
以蔬菜废弃物为原料的厌氧消化过程产甲烷能力下降时,通过添加微量元素可使其恢复稳定状态,因此研究微量元素对厌氧消化系统微生物结构的影响对优化系统性能具有重要意义.采用70 L厌氧发酵罐,有效体积59.5 L,在中温35℃条件下进行蔬菜废弃物厌氧消化的连续冲击负荷试验,根据CH_4含量变化规律,及时添加微量元素(Fe、Co、Ni)促进厌氧消化过程.样品采用16SrRNA基因扩增和MiSeq测序技术分析微生物群落的结构.结果表明,微量元素对细菌群落的影响主要作用于拟杆菌门、厚壁菌门及螺旋菌门.在属水平上,第一次微量元素的添加诱导了拟杆菌门中的VadinBC27 wastewater-sludge的增加,相对丰度从54.1%升至68%,降低了厚壁菌门中Erysipelotrichaceae UGG-004以及螺旋菌门中Sphaerochaeta.第二次微量元素的添加,主要降低了螺旋菌门中的Sphaerochaeta,相对丰度从11.4%到4.4%,以及诱导拟杆菌门中Bacteroides的产生,提高了原料利用率,降低了酸化的抑制作用.微量元素对蔬菜废弃物厌氧消化过程中产甲烷菌群落的影响主要在甲基营养型Candidatus Methanoplasma、甲烷鬃菌属为主导的乙酸营养型.当挥发性脂肪酸含量较高时,Candidatus Methanoplasma占主导地位,微量元素添加后则会诱导甲烷鬃菌为主导的乙酸营养型甲烷菌的产生,相对丰度从2.3%增至80%促进挥发性脂肪酸的消耗转化.本研究表明,微量元素的添加对于微生物群落结构的改变显著,促进厌氧消化过程水解酸化与甲烷化的平衡,从而稳定运行.
To optimize the performance of continuous anaerobic digestion of vegetable waste, the relationship between trace elements and microbial community structure was explored. In this study, the continuous anaerobic digestion(AD) of vegetable waste(VW) was carried out in the 70-L anaerobic reactors under mesophilic conditions(35 ℃), and the trace element supplementation was based on the CH_4 content in order to stabilize the process under high organic loading rate(OLR) for a long run-length. The microbial community structure was obtained through MiSeq sequencing of 16 S rRNA gene amplicons. For the bacterial communities, the addition of trace elements had significant effects on Bacteroidetes, Spirochaetae, and Firmicutes. At the genus level, the first trace elements addition induced VadinBC27 wastewater-sludge to increase from 54.1% to 68% and reduced Erysipelotrichaceae UGG-004 and Sphaerochaeta. The first trace element addition induced the abundance of Bacteroides to increase, reduced that of Sphaerochaeta from 11.4% to 4.4% to improve the utilization of raw materials, and reduced the inhibition of acidification. For the methanogen communities, the addition of trace elements had significant effects on Candidatus Methanoplasma, which is a methantrophicutilization methanogen and acetic-utilization methanogens with Methanosaeta dominated. Candidatus Methanoplasma dominated in the presence of high volatile fatty acids(VFA), and Methanosaeta dominated from 2.3% to 80% after the addition of trace elements to promote the consumption and conversion of VFA. Therefore, trace elements can promote the balance of hydrolysis and acidification and methanation in anaerobic digestion of vegetable waste, thus stabilizing the operation.
To optimize the performance of continuous anaerobic digestion of vegetable waste, the relationship between trace elements and microbial community structure was explored. In this study, the continuous anaerobic digestion(AD) of vegetable waste(VW) was carried out in the 70-L anaerobic reactors under mesophilic conditions(35 ℃), and the trace element supplementation was based on the CH_4 content in order to stabilize the process under high organic loading rate(OLR) for a long run-length. The microbial community structure was obtained through MiSeq sequencing of 16 S rRNA gene amplicons. For the bacterial communities, the addition of trace elements had significant effects on Bacteroidetes, Spirochaetae, and Firmicutes. At the genus level, the first trace elements addition induced VadinBC27 wastewater-sludge to increase from 54.1% to 68% and reduced Erysipelotrichaceae UGG-004 and Sphaerochaeta. The first trace element addition induced the abundance of Bacteroides to increase, reduced that of Sphaerochaeta from 11.4% to 4.4% to improve the utilization of raw materials, and reduced the inhibition of acidification. For the methanogen communities, the addition of trace elements had significant effects on Candidatus Methanoplasma, which is a methantrophicutilization methanogen and acetic-utilization methanogens with Methanosaeta dominated. Candidatus Methanoplasma dominated in the presence of high volatile fatty acids(VFA), and Methanosaeta dominated from 2.3% to 80% after the addition of trace elements to promote the consumption and conversion of VFA. Therefore, trace elements can promote the balance of hydrolysis and acidification and methanation in anaerobic digestion of vegetable waste, thus stabilizing the operation.
引文
1张丽颖,姜永海,邓舟,张妍.城市生物质废物厌氧消化处理可行性分析[J].环境科学与技术,2010, 33(5):143-146[Zhang LY,Jiang YH,Deng Z, Zhang Y. Feasibility analysis of treating and recycling municipal biomass waste using anaerobic digestion[J]. Environ Sci Technol, 2010,33(5):143-146]
2董永亮.果蔬废弃物两相厌氧消化特征研究[J].能源环境保护,2011,25(4):19-23, 28[Dong YL. Characteristics of two-phase anaerobic digestion of fruits and vegetables waste[J]. Energy Environ Protect.2011,25(4):19-23, 28]
3毛菁菁.蔬菜废弃物中温单相厌氧消化性能研究[D].北京:北京化工大学,2010[Mao JJ. Vegetable waste performances of onephase anaerobic digestion[D]. Beijing:Beijing University of Chemical Technology, 2010]
4刘广民,董永亮,薛建良,肖芳宇,尹莉莉.果蔬废弃物厌氧消化特征及固体减量研究[J].环境科学与技术,2009, 32(3):27-30, 49[Liu GM, Dong YL, Xue JL, Xiao FY, Yin LL. Characteristics of anaerobic digestion of fruit and vegetable waste and solid decrement[J]. Environ Sci Technol, 2009, 32(3):27-30, 49]
5贾传兴,彭绪亚,黄媛媛,李小风.有机垃圾厌氧消化系统失稳预警指标的研究进展[J].中国给水排水,2011,27(24):30-35[Jia CX,Peng XY, Huang YY, Li XF. Research progress in early-warning indicators for imbalance of anaerobic digestion system of organic wastes[J]. Chin Water Wastewater, 2011, 27(24):30-35]
6 Evranos B, Demirel B. The impact of Ni, Co and Mo supplementation on methane yield from anaerobic mono-digestion of maize silage[J].Environ Technol, 2015,36(12):1556-1562
7 Schmidt T, Nelles M, Scholwin F, Jurgen P. Trace element supplementation in the biogas production from wheat stillageoptimization of metal dosing[J]. Bioresour Technol, 2014, 168:80-85
8 Banks CJ, Zhang Y, Jiang Y, Heaven S. Trace element requirements for stable food waste digestion at elevated ammonia concentrations[J].Bioresour Technol, 2012, 104:127-135
9 Moested TJ, Nordell E, Yekta SS, Lundgren J, Marti M, Sundberg C.Effects of trace element addition on process stability during anaerobic co-digestion of OFMSW and slaughterhouse waste[J]. Waste Manage,2016, 47:11-20
10 Ferry JG. The chemical biology of methanogenesis[J]. Planet Space Sci,2010,58(14/15):1775-1783
11 Glass JB, Orphan VJ. Trace metal requirements for microbial enzymes involved in the production and consumption of methane and nitrous oxide[J]. Front Microbiol, 2012, 3(61):1-20
12 Hochheimer A, Hedderich R, Thauer RK. The formylmethanofuran dehydrogenase isoenzymes in methanobacterium wolfei and methanobacterium thermoautotrophicum:induction of the molybdenum isoenzyme by molybdate and constitutive synthesis of the tungsten isoenzyme[J]. Arch Microbiol, 1998, 170(5):389-393
13 Shima S, Warkentin E, Thauer RK, Ermler U. Structure and function of enzymes involved in the methanogenic pathway utilizing carbon dioxide and molecular hydrogen[J]. JBiosci Bioeng, 2002, 93(6):519-530
14李蕾,何琴,马垚.厌氧消化过程稳定性与微生物群落的相关惟[J].中国环境科学,2016, 36(11):3397-3404[Li L,He Q,MaY.Investigation on the relationship between process stability and microbial community in anaerobic digestion[J]. Chin Environ Sci, 2016, 36(11):3397-3404]
15 Ljungdhal LG. The autotrophic pathway of acetate synthesis in acetogenic bacteria[J]. Annu Rev Microbiol, 1986, 40:415-450
16 Jarvisa A, Nordberg A,Jarlsvik T,Mathisen B, Svensson BH.Improvement of a grass-clover silage-fed biogas process by the addition of cobalt[J]. Biomass Bioenerg, 1997, 12(6):453-460
17 Choong YY, Norli I, Abdullah AZ, Yhaya MF. Impacts of trace element supplementation on the performance of anaerobic digestion process:a critical review[J]. Bioresour Technol, 2016, 209:369-379
18 Goux X, Calusinska M, Lemaigre S, Marynowska M. Microbial community dynamics in replicate anaerobic digesters exposed sequentially to increasing organic loading rate, acidosis, and process recovery[J]. Biotechnol Biofuels, 2015, 8(1):1-18
19 RinconB, Borja R, Gonzalez JM, Portillo MC. Influence of organic loading rate and hydraulic retention time on the performance, stability and microbial communities of one-stage anaerobic digestion of twophase olive mill solid residue[J]. Biochem Eng J, 2008, 40(2):253-261
20 Guo XH, Cheng W, Sun FQ, Zhu WJ. A comparison of microbial characteristics between the thermophilic and mesophilic anaerobic digesters exposed to elevated food waste loadings[J]. Bioresour Technol, 2014, 152(152C):420-428
21 Jaenicke S, Ander C, Bekel T, Bisdorf R, Droge M. Comparative and joint analysis of two metagenomics datasets from a biogas fermenter obtained by 454-pyrosequencing[J]. PLoS ONE, 2011, 6(1):e14519
22李秋敏,吴燕,董明华.云南热带户用沼气地的原核生物群落结构研究[J].微生物学通报.2015, 42(1):54-63[Li QM,Wu Y,Dong MH.Prokaryotic community structure of household biogas digesters in the tropical climate zones of Yunnan[J]. Microbiol Chin, 2015, 42(1):54-63]
23 Rautio M, Eerola E, Vaisanentunkelrott ML, Molitoris D, Lawson P,Collins MD. Reclassification of bacteroides putredinis(Weinberg et al.1937)in a new genus Alistipes gen. nov. as Alistipes putredinis comb.nov. and description of Alistipes finegoldii sp. nov. from human sources[J]. Syst Appl Microbiol, 2003, 26(2):182-188
24 Greiner T, Backhed F. Effects of the gut microbiota on obesity anf glucose homeostasis[J]. Trends Endocrin Met, 2011, 22(4):117-123
25 Li A, Chu Y, Wang XM, Ren LF, Yu J, Liu XL. A pyrosequencing-based metagenomics study of methane-producing microbial community in solid-state biogas reactor[J]. Biotechnol Biofuels, 2013, 6(1):1-17
26 Lang K, Schuldes J, Klingl A, Klingl A, Poehlein A, Daniel R, Brune A.New mode of energy metabolism in the seventh order of methanogens as revealed by comparative genome analysis of"Candidatus Methanoplasma termitun"[J]. Appl Environ Microbiol, 2015, 81(4):1388-1352
27 Christy PM, Gopinath LR, Divya D. A review on anaerobic decomposition and enhancement of biogas production through enzymes and microorganisms[J]. Renew Sust Energ Rev, 2014, 34(6):167-173
2董永亮.果蔬废弃物两相厌氧消化特征研究[J].能源环境保护,2011,25(4):19-23, 28[Dong YL. Characteristics of two-phase anaerobic digestion of fruits and vegetables waste[J]. Energy Environ Protect.2011,25(4):19-23, 28]
3毛菁菁.蔬菜废弃物中温单相厌氧消化性能研究[D].北京:北京化工大学,2010[Mao JJ. Vegetable waste performances of onephase anaerobic digestion[D]. Beijing:Beijing University of Chemical Technology, 2010]
4刘广民,董永亮,薛建良,肖芳宇,尹莉莉.果蔬废弃物厌氧消化特征及固体减量研究[J].环境科学与技术,2009, 32(3):27-30, 49[Liu GM, Dong YL, Xue JL, Xiao FY, Yin LL. Characteristics of anaerobic digestion of fruit and vegetable waste and solid decrement[J]. Environ Sci Technol, 2009, 32(3):27-30, 49]
5贾传兴,彭绪亚,黄媛媛,李小风.有机垃圾厌氧消化系统失稳预警指标的研究进展[J].中国给水排水,2011,27(24):30-35[Jia CX,Peng XY, Huang YY, Li XF. Research progress in early-warning indicators for imbalance of anaerobic digestion system of organic wastes[J]. Chin Water Wastewater, 2011, 27(24):30-35]
6 Evranos B, Demirel B. The impact of Ni, Co and Mo supplementation on methane yield from anaerobic mono-digestion of maize silage[J].Environ Technol, 2015,36(12):1556-1562
7 Schmidt T, Nelles M, Scholwin F, Jurgen P. Trace element supplementation in the biogas production from wheat stillageoptimization of metal dosing[J]. Bioresour Technol, 2014, 168:80-85
8 Banks CJ, Zhang Y, Jiang Y, Heaven S. Trace element requirements for stable food waste digestion at elevated ammonia concentrations[J].Bioresour Technol, 2012, 104:127-135
9 Moested TJ, Nordell E, Yekta SS, Lundgren J, Marti M, Sundberg C.Effects of trace element addition on process stability during anaerobic co-digestion of OFMSW and slaughterhouse waste[J]. Waste Manage,2016, 47:11-20
10 Ferry JG. The chemical biology of methanogenesis[J]. Planet Space Sci,2010,58(14/15):1775-1783
11 Glass JB, Orphan VJ. Trace metal requirements for microbial enzymes involved in the production and consumption of methane and nitrous oxide[J]. Front Microbiol, 2012, 3(61):1-20
12 Hochheimer A, Hedderich R, Thauer RK. The formylmethanofuran dehydrogenase isoenzymes in methanobacterium wolfei and methanobacterium thermoautotrophicum:induction of the molybdenum isoenzyme by molybdate and constitutive synthesis of the tungsten isoenzyme[J]. Arch Microbiol, 1998, 170(5):389-393
13 Shima S, Warkentin E, Thauer RK, Ermler U. Structure and function of enzymes involved in the methanogenic pathway utilizing carbon dioxide and molecular hydrogen[J]. JBiosci Bioeng, 2002, 93(6):519-530
14李蕾,何琴,马垚.厌氧消化过程稳定性与微生物群落的相关惟[J].中国环境科学,2016, 36(11):3397-3404[Li L,He Q,MaY.Investigation on the relationship between process stability and microbial community in anaerobic digestion[J]. Chin Environ Sci, 2016, 36(11):3397-3404]
15 Ljungdhal LG. The autotrophic pathway of acetate synthesis in acetogenic bacteria[J]. Annu Rev Microbiol, 1986, 40:415-450
16 Jarvisa A, Nordberg A,Jarlsvik T,Mathisen B, Svensson BH.Improvement of a grass-clover silage-fed biogas process by the addition of cobalt[J]. Biomass Bioenerg, 1997, 12(6):453-460
17 Choong YY, Norli I, Abdullah AZ, Yhaya MF. Impacts of trace element supplementation on the performance of anaerobic digestion process:a critical review[J]. Bioresour Technol, 2016, 209:369-379
18 Goux X, Calusinska M, Lemaigre S, Marynowska M. Microbial community dynamics in replicate anaerobic digesters exposed sequentially to increasing organic loading rate, acidosis, and process recovery[J]. Biotechnol Biofuels, 2015, 8(1):1-18
19 RinconB, Borja R, Gonzalez JM, Portillo MC. Influence of organic loading rate and hydraulic retention time on the performance, stability and microbial communities of one-stage anaerobic digestion of twophase olive mill solid residue[J]. Biochem Eng J, 2008, 40(2):253-261
20 Guo XH, Cheng W, Sun FQ, Zhu WJ. A comparison of microbial characteristics between the thermophilic and mesophilic anaerobic digesters exposed to elevated food waste loadings[J]. Bioresour Technol, 2014, 152(152C):420-428
21 Jaenicke S, Ander C, Bekel T, Bisdorf R, Droge M. Comparative and joint analysis of two metagenomics datasets from a biogas fermenter obtained by 454-pyrosequencing[J]. PLoS ONE, 2011, 6(1):e14519
22李秋敏,吴燕,董明华.云南热带户用沼气地的原核生物群落结构研究[J].微生物学通报.2015, 42(1):54-63[Li QM,Wu Y,Dong MH.Prokaryotic community structure of household biogas digesters in the tropical climate zones of Yunnan[J]. Microbiol Chin, 2015, 42(1):54-63]
23 Rautio M, Eerola E, Vaisanentunkelrott ML, Molitoris D, Lawson P,Collins MD. Reclassification of bacteroides putredinis(Weinberg et al.1937)in a new genus Alistipes gen. nov. as Alistipes putredinis comb.nov. and description of Alistipes finegoldii sp. nov. from human sources[J]. Syst Appl Microbiol, 2003, 26(2):182-188
24 Greiner T, Backhed F. Effects of the gut microbiota on obesity anf glucose homeostasis[J]. Trends Endocrin Met, 2011, 22(4):117-123
25 Li A, Chu Y, Wang XM, Ren LF, Yu J, Liu XL. A pyrosequencing-based metagenomics study of methane-producing microbial community in solid-state biogas reactor[J]. Biotechnol Biofuels, 2013, 6(1):1-17
26 Lang K, Schuldes J, Klingl A, Klingl A, Poehlein A, Daniel R, Brune A.New mode of energy metabolism in the seventh order of methanogens as revealed by comparative genome analysis of"Candidatus Methanoplasma termitun"[J]. Appl Environ Microbiol, 2015, 81(4):1388-1352
27 Christy PM, Gopinath LR, Divya D. A review on anaerobic decomposition and enhancement of biogas production through enzymes and microorganisms[J]. Renew Sust Energ Rev, 2014, 34(6):167-173