亚硝酸盐型厌氧甲烷氧化过程强化新视角:排泥及其微生物机制研究
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摘要
亚硝酸盐型厌氧甲烷氧化(Nitrite-dependent anaerobic methane oxidation, n-DAMO)是微生物在厌氧条件下利用甲烷还原亚硝酸盐的过程.本研究通过排泥的策略对n-DAMO过程进行强化,并比较分析了反应器中微生物的群落结构及功能微生物数量.结果发现,与对照组相比,排泥后实验组反应器的脱氮速率从17.00 mg·L~(-1)·d~(-1)提高到73.10 mg·L~(-1)·d~(-1).排泥后反应器中n-DAMO细菌的相对丰度从38.3%上升到67.7%,功能微生物的基因拷贝数由1.404×10~8 copies·g~(-1)增长到4.854×10~8 copies·g~(-1),污泥比活性提高了2.95倍.与之相反,初始反应器中其余优势微生物Unclassified_GCA004、Unclassified_Rhodocyclaceae、Unclassified_Fimbriimonadaceae与Methylosinu相对丰度分别下降为原来的27.66%、32.65%、4.35%、20.27%.结果表明,排泥可以有效地强化n-DAMO过程,同时促进功能微生物的生长,主要原因在于排泥排出了非目标微生物,使得目标微生物大量生长.本研究为强化n-DAMO过程及加快n-DAMO微生物的富集提供了一条新思路,并为进一步推动n-DAMO过程的工程应用提供了理论基础.
Specific microorganisms involving in nitrite-dependent anaerobic methane oxidation(n-DAMO) process could utilize methane to reduce nitrite under anaerobic condition. In this study, the effect of withdrawing sludge on n-DAMO process was investigated and its microorganism community as well as functional microorganism abundance were analyzed. Compared with control group, the nitrogen removal rate of experimental group increased from 17.00 mg·L~(-1)·d~(-1) to 73.10 mg·L~(-1)·d~(-1) significantly after withdrawing sludge. Microorganism community analysis showed that the relative abundance of n-DAMO bacteria increased from 38.3% to 67.7% after withdrawing sludge. Functional microorganism abundance increased from 1.404×10~8 copies·g~(-1) to 4.854×10~8 copies·g~(-1) and sludge specific activity increased by 295%. In contrast, the relative abundances of unclassified_GCA004, unclassified_Rhodocyclacea,unclassified_Fimbriimonadaceae and Methylosinu decreased to 27.66%, 32.65%, 4.35% and 20.27%, respectively, after withdrawing sludge. The results indicate that withdrawing sludge can strengthen n-DAMO process and promote functional microorganism growth, which might be due to the increase of n-DAMO functional microorganisms abundance in the system. This study gives a new insight to strengthen n-DAMO process and accelerate n-DAMO microorganism enrichment, which provides theoretical basis for the application of n-DAMO process.
Specific microorganisms involving in nitrite-dependent anaerobic methane oxidation(n-DAMO) process could utilize methane to reduce nitrite under anaerobic condition. In this study, the effect of withdrawing sludge on n-DAMO process was investigated and its microorganism community as well as functional microorganism abundance were analyzed. Compared with control group, the nitrogen removal rate of experimental group increased from 17.00 mg·L~(-1)·d~(-1) to 73.10 mg·L~(-1)·d~(-1) significantly after withdrawing sludge. Microorganism community analysis showed that the relative abundance of n-DAMO bacteria increased from 38.3% to 67.7% after withdrawing sludge. Functional microorganism abundance increased from 1.404×10~8 copies·g~(-1) to 4.854×10~8 copies·g~(-1) and sludge specific activity increased by 295%. In contrast, the relative abundances of unclassified_GCA004, unclassified_Rhodocyclacea,unclassified_Fimbriimonadaceae and Methylosinu decreased to 27.66%, 32.65%, 4.35% and 20.27%, respectively, after withdrawing sludge. The results indicate that withdrawing sludge can strengthen n-DAMO process and promote functional microorganism growth, which might be due to the increase of n-DAMO functional microorganisms abundance in the system. This study gives a new insight to strengthen n-DAMO process and accelerate n-DAMO microorganism enrichment, which provides theoretical basis for the application of n-DAMO process.
引文
Allegue T,Arias A,Fernandez-Gonzalez N,et al.2018.Enrichment of nitrite-dependent anaerobic methane oxidizing bacteria in a membrane bioreactor[J].Chemical Engineering Journal,347:721-730
柴风光,卢培利,李微薇,等.2018.利用硝酸盐和亚硝酸盐同步富集厌氧甲烷氧化微生物的比较实验[J].微生物学通报,45(4):762-770
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Ettwig K F,Shima S,Pas-Schoonen K T V D,et al.2008.Denitrifying bacteria anaerobically oxidize methane in the absence of Archaea[J].Environmental Microbiology,10(11):3164-3173
Ettwig K F,Van A T,Kt P S,et al.2009.Enrichment and molecular detection of denitrifying methanotrophic bacteria of the NC10 phylum[J].Applied and Environmental Microbiology,75(11):3656-3662
Fernandes S O,Lokabharathi P A,Bonin P C,et al.2010.Denitrification:an important pathway for nitrous oxide production in tropical mangrove sediments (Goa,India)[J].Journal of Environmental Quality,39(4):1507-1516
Fu L,Ding J,Lu Y Z,et al.2017.Nitrogen source effects on the denitrifying anaerobic methane oxidation culture and anaerobic ammonium oxidation bacteria enrichment process[J].Applied Microbiology and Biotechnology,101(9):3895-3906
Han P,Gu J D.2013.A newly designed degenerate PCR primer based on pmoA,gene for detection of nitrite-dependent anaerobic methane-oxidizing bacteria from different ecological niches[J].Applied Microbiology and Biotechnology,97(23):10155-10162
He Z,Geng S,Shen L,et al.2015a.The short- and long-term effects of environmental conditions on anaerobic methane oxidation coupled to nitrite reduction[J].Water Research,68(47):554-562
He Z,Cai C,Shen L,et al.2015b.Effect of inoculum sources on the enrichment of nitrite-dependent anaerobic methane-oxidizing bacteria[J].Applied Microbiology and Biotechnology,99(2):939-946
He Z,Geng S,Wang L,et al.2016.Improvement of mineral nutrient concentrations and pH control for the nitrite-dependent anaerobic methane oxidation process[J].Separation and Purification Technology,162(3):148-153
Hu B L,Shen L D,Huang Q,et al.2014a.Evidence for nitrite-dependent anaerobic methane oxidation as a previously overlooked microbial methane sink in wetlands[J].Proceedings of the National Academy of Sciences of the United States of America,111(12):4495-4500
Hu B,He Z,Geng S,et al.2014b.Cultivation of nitrite-dependent anaerobic methane-oxidizing bacteria:impact of reactor configuration[J].Applied Microbiology and Biotechnology,98(18):7983-7991
Hu S,Zeng R J,Burow L C,et al.2009.Enrichment of denitrifying anaerobic methane oxidizing microorganisms[J].Environmental Microbiology Reports,1(5):377-384
Kampman C,Hendrickx T L G,Luesken F A,et al.2012.Enrichment of denitrifying methanotrophic bacteria for application after direct low-temperature anaerobic sewage treatment[J].Journal of Hazardous Materials,227:164-171
Kampman C,Temmink H,Hendrickx T L G,et al.2014.Enrichment of denitrifying methanotrophic bacteria from municipal wastewater sludge in a membrane bioreactor at 20℃[J].Journal of Hazardous Materials,274:428-435
Li W,Lu P,Chai F,et al.2018.Long-term nitrate removal through methane-dependent denitrification microorganisms in sequencing batch reactors fed with only nitrate and methane[J].Amb Express,8(1):108
Lieberman R L,Rosenzweig A C.2004.Biological methane oxidation:Regulation,biochemistry,and active site structure of particulate methane monooxygenase[J].Critical Reviews in Biochemistry and Molecular Biology,39(3):147-164
Liu J,Sun F,Wang L,et al.2013.Molecular characterization of a microbial consortium involved in methane oxidation coupled to denitrification under micro-aerobic conditions[J].Microbial Biotechnology,7(1):64-76
Lu P,Liu T,Ni B,et al.2019.Growth kinetics of Candidatus ‘Methanoperedens nitroreducens' enriched in a laboratory reactor[J].Science of the Total Environment,695:442-450
Lu Y Z,Fu L,Ding J,et al.2016.Cr(VI) reduction coupled with anaerobic oxidation of methane in a laboratory reactor[J].Water Research,102:445-452
Luesken F A,Jaime Sánchez,Alen T A,et al.2011.Simultaneous nitrite-dependent anaerobic methane and ammonium oxidation processes[J].Applied and Environmental Microbiology,77(19):6802-6807
Ordaz A,López J C,Figueroa-González I,et al.2014.Assessment of methane biodegradation kinetics in two-phase partitioning bioreactors by pulse respirometry[J].Water Research,67:46-54
Raghoebarsing A A,Pol A,Pas-Schoonen K T V D,et al.2006.A microbial consortium couples anaerobic methane oxidation to denitrification[J].Nature,440(7086):918-921
沈李东,胡宝兰,郑平.2011.甲烷厌氧氧化微生物的研究进展[J].土壤学报,48(3):619-628
Solomon S,Qin M,Manning Z,et al.2007.Climate Change 2007:The Physical Science Basis.Working Group I Contribution to the Fourth Assessment Report of the IPCC[M].Cambridge University Press.1-21
Wan Y,Ruan X,Zhang Y,et al.2017.Illumina sequencing-based analysis of sediment bacteria community in different trophic status freshwater lakes[J].Microbiology Open,6:e450.DOI:10.1002/mbo3.450
Wang R,Han X,Lu P,et al.2016.Co-occurrence and diversity of nitrite-dependent anaerobic methane oxidizing and anaerobic ammonium oxidizing bacteria in the water-level fluctuation zone of the three gorges reservoir,China[J].Fresenius Environmental Bulletin,25(12):5456-5466
Wu Q,Chang J,Yan X,et al.2016a.Electrical stimulation enhanced denitrification of nitrite-dependent anaerobic methane-oxidizing bacteria[J].Biochemical Engineering Journal,106:125-128
Wu P,Xiong X,Xu Z,et al.2016b.Bacterial communities in the rhizospheres of three mangrove tree species from Beilun Estuary,China[J].Plos One,11(10):e0164082.DOI:10.1371/journal.pone.0164082
Yoshida M,Ishii S,Fujii D,et al.2012.Identification of active denitrifiers in rice paddy soil by DNA- and RNA-based analyses[J].Microbes & Environments,27(4):456-461
Yue X,Yu G,Liu Z,et al.2018.Start-up of the completely autotrophic nitrogen removal over nitrite process with a submerged aerated biological filter and the effect of inorganic carbon on nitrogen removal and microbial activity[J].Bioresource Technology,254:347-352
赵荣,朱雷,吴箐,等.2017.亚硝酸盐型甲烷厌氧氧化过程影响因素研究[J].环境科学学报,37(1):178-184
Zhou L,Wang Y,Long X E,et al.2014.High abundance and diversity of nitrite-dependent anaerobic methane-oxidizing bacteria in a paddy field profile[J].FEMS Microbiology Letters,360(1):33-41
Zhu B,Van G D,Fritz C,et al.2012.Anaerobic oxidization of methane in a minerotrophic peatland:enrichment of nitrite-dependent methane-oxidizing bacteria[J].Applied and Environmental Microbiology,78(24):8657-8665
柴风光,卢培利,李微薇,等.2018.利用硝酸盐和亚硝酸盐同步富集厌氧甲烷氧化微生物的比较实验[J].微生物学通报,45(4):762-770
Chen J,Zhou Z C,Gu J D.2014.Occurrence and diversity of nitrite-dependent anaerobic methane oxidation bacteria in the sediments of the South China Sea revealed by amplification of both 16S rRNA and pmoA genes[J].Applied Microbiology and Biotechnology,98(12):5685-5696
Ettwig K F,Shima S,Pas-Schoonen K T V D,et al.2008.Denitrifying bacteria anaerobically oxidize methane in the absence of Archaea[J].Environmental Microbiology,10(11):3164-3173
Ettwig K F,Van A T,Kt P S,et al.2009.Enrichment and molecular detection of denitrifying methanotrophic bacteria of the NC10 phylum[J].Applied and Environmental Microbiology,75(11):3656-3662
Fernandes S O,Lokabharathi P A,Bonin P C,et al.2010.Denitrification:an important pathway for nitrous oxide production in tropical mangrove sediments (Goa,India)[J].Journal of Environmental Quality,39(4):1507-1516
Fu L,Ding J,Lu Y Z,et al.2017.Nitrogen source effects on the denitrifying anaerobic methane oxidation culture and anaerobic ammonium oxidation bacteria enrichment process[J].Applied Microbiology and Biotechnology,101(9):3895-3906
Han P,Gu J D.2013.A newly designed degenerate PCR primer based on pmoA,gene for detection of nitrite-dependent anaerobic methane-oxidizing bacteria from different ecological niches[J].Applied Microbiology and Biotechnology,97(23):10155-10162
He Z,Geng S,Shen L,et al.2015a.The short- and long-term effects of environmental conditions on anaerobic methane oxidation coupled to nitrite reduction[J].Water Research,68(47):554-562
He Z,Cai C,Shen L,et al.2015b.Effect of inoculum sources on the enrichment of nitrite-dependent anaerobic methane-oxidizing bacteria[J].Applied Microbiology and Biotechnology,99(2):939-946
He Z,Geng S,Wang L,et al.2016.Improvement of mineral nutrient concentrations and pH control for the nitrite-dependent anaerobic methane oxidation process[J].Separation and Purification Technology,162(3):148-153
Hu B L,Shen L D,Huang Q,et al.2014a.Evidence for nitrite-dependent anaerobic methane oxidation as a previously overlooked microbial methane sink in wetlands[J].Proceedings of the National Academy of Sciences of the United States of America,111(12):4495-4500
Hu B,He Z,Geng S,et al.2014b.Cultivation of nitrite-dependent anaerobic methane-oxidizing bacteria:impact of reactor configuration[J].Applied Microbiology and Biotechnology,98(18):7983-7991
Hu S,Zeng R J,Burow L C,et al.2009.Enrichment of denitrifying anaerobic methane oxidizing microorganisms[J].Environmental Microbiology Reports,1(5):377-384
Kampman C,Hendrickx T L G,Luesken F A,et al.2012.Enrichment of denitrifying methanotrophic bacteria for application after direct low-temperature anaerobic sewage treatment[J].Journal of Hazardous Materials,227:164-171
Kampman C,Temmink H,Hendrickx T L G,et al.2014.Enrichment of denitrifying methanotrophic bacteria from municipal wastewater sludge in a membrane bioreactor at 20℃[J].Journal of Hazardous Materials,274:428-435
Li W,Lu P,Chai F,et al.2018.Long-term nitrate removal through methane-dependent denitrification microorganisms in sequencing batch reactors fed with only nitrate and methane[J].Amb Express,8(1):108
Lieberman R L,Rosenzweig A C.2004.Biological methane oxidation:Regulation,biochemistry,and active site structure of particulate methane monooxygenase[J].Critical Reviews in Biochemistry and Molecular Biology,39(3):147-164
Liu J,Sun F,Wang L,et al.2013.Molecular characterization of a microbial consortium involved in methane oxidation coupled to denitrification under micro-aerobic conditions[J].Microbial Biotechnology,7(1):64-76
Lu P,Liu T,Ni B,et al.2019.Growth kinetics of Candidatus ‘Methanoperedens nitroreducens' enriched in a laboratory reactor[J].Science of the Total Environment,695:442-450
Lu Y Z,Fu L,Ding J,et al.2016.Cr(VI) reduction coupled with anaerobic oxidation of methane in a laboratory reactor[J].Water Research,102:445-452
Luesken F A,Jaime Sánchez,Alen T A,et al.2011.Simultaneous nitrite-dependent anaerobic methane and ammonium oxidation processes[J].Applied and Environmental Microbiology,77(19):6802-6807
Ordaz A,López J C,Figueroa-González I,et al.2014.Assessment of methane biodegradation kinetics in two-phase partitioning bioreactors by pulse respirometry[J].Water Research,67:46-54
Raghoebarsing A A,Pol A,Pas-Schoonen K T V D,et al.2006.A microbial consortium couples anaerobic methane oxidation to denitrification[J].Nature,440(7086):918-921
沈李东,胡宝兰,郑平.2011.甲烷厌氧氧化微生物的研究进展[J].土壤学报,48(3):619-628
Solomon S,Qin M,Manning Z,et al.2007.Climate Change 2007:The Physical Science Basis.Working Group I Contribution to the Fourth Assessment Report of the IPCC[M].Cambridge University Press.1-21
Wan Y,Ruan X,Zhang Y,et al.2017.Illumina sequencing-based analysis of sediment bacteria community in different trophic status freshwater lakes[J].Microbiology Open,6:e450.DOI:10.1002/mbo3.450
Wang R,Han X,Lu P,et al.2016.Co-occurrence and diversity of nitrite-dependent anaerobic methane oxidizing and anaerobic ammonium oxidizing bacteria in the water-level fluctuation zone of the three gorges reservoir,China[J].Fresenius Environmental Bulletin,25(12):5456-5466
Wu Q,Chang J,Yan X,et al.2016a.Electrical stimulation enhanced denitrification of nitrite-dependent anaerobic methane-oxidizing bacteria[J].Biochemical Engineering Journal,106:125-128
Wu P,Xiong X,Xu Z,et al.2016b.Bacterial communities in the rhizospheres of three mangrove tree species from Beilun Estuary,China[J].Plos One,11(10):e0164082.DOI:10.1371/journal.pone.0164082
Yoshida M,Ishii S,Fujii D,et al.2012.Identification of active denitrifiers in rice paddy soil by DNA- and RNA-based analyses[J].Microbes & Environments,27(4):456-461
Yue X,Yu G,Liu Z,et al.2018.Start-up of the completely autotrophic nitrogen removal over nitrite process with a submerged aerated biological filter and the effect of inorganic carbon on nitrogen removal and microbial activity[J].Bioresource Technology,254:347-352
赵荣,朱雷,吴箐,等.2017.亚硝酸盐型甲烷厌氧氧化过程影响因素研究[J].环境科学学报,37(1):178-184
Zhou L,Wang Y,Long X E,et al.2014.High abundance and diversity of nitrite-dependent anaerobic methane-oxidizing bacteria in a paddy field profile[J].FEMS Microbiology Letters,360(1):33-41
Zhu B,Van G D,Fritz C,et al.2012.Anaerobic oxidization of methane in a minerotrophic peatland:enrichment of nitrite-dependent methane-oxidizing bacteria[J].Applied and Environmental Microbiology,78(24):8657-8665
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