不同溶解氧浓度下硝化工艺中微生物种群结构对比
|
摘要
运用高通量测序技术分析了不同溶解氧(DO)浓度下硝化工艺微生物种群结构.结果表明,低氧硝化反应器(R_L,DO浓度为0. 2～0. 3 mg·L~(-1))比高氧硝化反应器[R_H,DO=(2. 0±0. 1) mg·L~(-1)]具有更高的种群多样性,而R_H中微生物种群功能组织性更高.尽管R_H和R_L共有物种信息达85%以上,但DO浓度的不同造成了单个物种在相对丰度上的显著差异.Proteobacteria门(80. 7%)在R_H中被高度富集,其中Nitrosomonas菌属相对丰度达到65. 1%;而在R_L中则以Proteobacteria(43. 8%)、Firmicutes (20. 0%)以及Bacteroidetes (15. 1%)为共同主导者,同时R_L中含有大量Lactococcus、Anaerolineaceae以及Rhodocyclaceae等可在厌氧或缺氧条件下进行水解发酵作用的菌属. Nitrosomonas oligotropha和Nitrosomonas europaea分别为RH和RL中优势氨氧化细菌,而亚硝酸盐氧化细菌均以Nitrospira defluvii为主导.反应器中NH_4~+-N和NO_2~--N浓度(而非DO浓度)是上述硝化菌群被选择性富集的关键因素.
High-throughput sequencing was applied to analyze the microbial community structure of nitrifying reactors operated with different dissolved oxygen( DO) levels. Results showed that the nitrifying reactor( R_L) run with low DO( 0. 2-0. 3 mg·L~(-1)) exhibited greater microbial richness and diversity than the reactor run under the high DO condition [R_H,DO =( 2. 0 ± 0. 1) mg·L~(-1)]. In contrast,the microbial community in R_H was more highly functionally organized than that in R_L. Although the communities in R_H and R_L shared over 85% of the total sampled genetic information,the relative abundance of some individual species varied between the different DO conditions. Members of the Proteobacteria phylum,which accounted for 80. 7% of the total microbes in R_H,were highly enriched,and the relative abundance of Nitrosomonas reached to 65. 1%. However,the microbial community in R_L was dominated by Proteobacteria( 43. 8%),Firmicutes( 20. 0%),and Bacteroidetes( 15. 1%). In addition,a large fraction of bacteria possessing hydrolyzation and fermentation functions under anaerobic or anoxic conditions were also present in R_L including Lactococcus,Anaerolineaceae,and Rhodocyclaceae. As known ammonium-oxidizing bacteria,Nitrosomonas oligotropha and Nitrosomonas europaea were enriched in the R_H and R_L,respectively,while Nitrospira defluvii,being a nitrite-oxidizing bacteria,dominated both reactors.Rather than DO,ammonia and nitrite availability should be key factors in the selective enrichment of these nitrifiers.
High-throughput sequencing was applied to analyze the microbial community structure of nitrifying reactors operated with different dissolved oxygen( DO) levels. Results showed that the nitrifying reactor( R_L) run with low DO( 0. 2-0. 3 mg·L~(-1)) exhibited greater microbial richness and diversity than the reactor run under the high DO condition [R_H,DO =( 2. 0 ± 0. 1) mg·L~(-1)]. In contrast,the microbial community in R_H was more highly functionally organized than that in R_L. Although the communities in R_H and R_L shared over 85% of the total sampled genetic information,the relative abundance of some individual species varied between the different DO conditions. Members of the Proteobacteria phylum,which accounted for 80. 7% of the total microbes in R_H,were highly enriched,and the relative abundance of Nitrosomonas reached to 65. 1%. However,the microbial community in R_L was dominated by Proteobacteria( 43. 8%),Firmicutes( 20. 0%),and Bacteroidetes( 15. 1%). In addition,a large fraction of bacteria possessing hydrolyzation and fermentation functions under anaerobic or anoxic conditions were also present in R_L including Lactococcus,Anaerolineaceae,and Rhodocyclaceae. As known ammonium-oxidizing bacteria,Nitrosomonas oligotropha and Nitrosomonas europaea were enriched in the R_H and R_L,respectively,while Nitrospira defluvii,being a nitrite-oxidizing bacteria,dominated both reactors.Rather than DO,ammonia and nitrite availability should be key factors in the selective enrichment of these nitrifiers.
引文
[1]Mc Carty P L,Bae J,Kim J.Domestic wastewater treatment as a net energy producer-Can this be achieved[J].Environmental Science&Technology,2011,45(17):7100-7106.
[2]张自杰,林荣忱,金儒霖.排水工程下册[M].(第四版).北京:中国建筑工业出版社,2000.
[3]Arnaldos M,Pagilla K R.Implementation of a demand-side approach to reduce aeration requirements of activated sludge systems:directed acclimation of biomass and its effect at the process level[J].Water Research,2014,62:147-155.
[4]Metcalf&Eddy.Wastewater engineering:treatment and reuse(4th ed.)[M].Boston:Mc Graw-Hill,2003.
[5]Abbassi B,Dullstein S,Rbiger N.Minimization of excess sludge production by increase of oxygen concentration in activated sludge flocs;Experimental and theoretical approach[J].Water Research,2000,34(1):139-146.
[6]Liu G Q,Wang J M.Long-term low do enriches and shifts nitrifier community in activated sludge[J].Environmental Science&Technology,2013,47(10):5109-5117.
[7]Fitzgerald C M,Camejo P,Oshlag J Z,et al.Ammoniaoxidizing microbial communities in reactors with efficient nitrification at low-dissolved oxygen[J].Water Research,2015,70:38-51.
[8]Park H D,Noguera D R.Evaluating the effect of dissolved oxygen on ammonia-oxidizing bacterial communities in activated sludge[J].Water Research,2004,38(14-15):3275-3286.
[9]Arnaldos M,Kunkel S A,Stark B C,et al.Characterization of heme protein expressed by ammonia-oxidizing bacteria under low dissolved oxygen conditions[J].Applied Microbiology and Biotechnology,2014,98(7):3231-3239.
[10]Bellucci M,Ofiteru I D,Graham D W,et al.Low-dissolvedoxygen nitrifying systems exploit ammonia-oxidizing bacteria with unusually high yields[J].Applied and Environmental Microbiology,2011,77(21):7787-7796.
[11]Wagner M,Loy A.Bacterial community composition and function in sewage treatment systems[J].Current Opinion in Biotechnology,2002,13(3):218-227.
[12]Ju F,Zhang T.Bacterial assembly and temporal dynamics in activated sludge of a full-scale municipal wastewater treatment plant[J].The ISME Journal,2015,9(3):683-695.
[13]Girvan M S,Campbell C D,Killham K,et al.Bacterial diversity promotes community stability and functional resilience after perturbation[J].Environmental Microbiology,2005,7(3):301-313.
[14]Briones A,Raskin L.Diversity and dynamics of microbial communities in engineered environments and their implications for process stability[J].Current Opinion in Biotechnology,2003,14(3):270-276.
[15]Wittebolle L,Marzorati M,Clement L,et al.Initial community evenness favours functionality under selective stress[J].Nature,2009,458(7238):623-626.
[16]van de Graaf A A,de Bruijn P,Robertson L A,et al.Autotrophic growth of anaerobic ammonium-oxidizing microorganisms in a fluidized bed reactor[J].Microbiology,1996,142(8):2187-2196.
[17]国家环境保护总局.水和废水监测分析方法[M].(第四版).北京:中国环境科学出版社,2002.
[18]Liu W R,Yang D H,Chen W J,et al.High-throughput sequencing-based microbial characterization of size fractionated biomass in an anoxic anammox reactor for low-strength wastewater at low temperatures[J].Bioresource Technology,2017,231:45-52.
[19]Manser R,Gujer W,Siegrist H.Consequences of mass transfer effects on the kinetics of nitrifiers[J].Water Research,2005,39(19):4633-4642.
[20]Arnaldos M,Amerlinck Y,Rehman U,et al.From the affinity constant to the half-saturation index:understanding conventional modeling concepts in novel wastewater treatment processes[J].Water Research,2015,70:458-470.
[21]王中玮,彭永臻,王淑莹,等.不同运行方式下低溶解氧污泥微膨胀的可行性研究[J].环境科学,2011,32(8):2347-2352.Wang Z W,Peng Y Z,Wang S Y,et al.Feasibility study for limited filamentous bulking under low dissolved oxygen at different operation regimes[J].Environmental Science,2011,32(8):2347-2352.
[22]Ma J X,Wang Z W,Yang Y,et al.Correlating microbial community structure and composition with aeration intensity in submerged membrane bioreactors by 454 high-throughput pyrosequencing[J].Water Research,2013,47(2):859-869.
[23]Gao D W,Fu Y,Tao Y,et al.Linking microbial community structure to membrane biofouling associated with varying dissolved oxygen concentrations[J].Bioresource Technology,2011,102(10):5626-5633.
[24]Marzorati M,Wittebolle L,Boon N,et al.How to get more out of molecular fingerprints:practical tools for microbial ecology[J].Environmental Microbiology,2008,10(6):1571-1581.
[25]Dytczak M A,Londry K L,Oleszkiewicz J A.Activated sludge operational regime has significant impact on the type of nitrifying community and its nitrification rates[J].Water Research,2008,42(8-9):2320-2328.
[26]Bollmann A,Br-Gilissen M J,Laanbroek H J.Growth at low ammonium concentrations and starvation response as potential factors involved in niche differentiation among ammonia-oxidizing bacteria[J].Applied and Environmental Microbiology,2002,68(10):4751-4757.
[27]Bollmann A,Laanbroek H J.Continuous culture enrichments of ammonia-oxidizing bacteria at low ammonium concentrations[J].FEMS Microbiology Ecology,2001,37(3):211-221.
[28]Nowka B,Daims H,Spieck E.Comparison of oxidation kinetics of nitrite-oxidizing bacteria:nitrite availability as a key factor in niche differentiation[J].Applied and Environmental Microbiology,2015,81(2):745-753.
[29]Park M R,Park H,Chandran K.Molecular and kinetic characterization of Planktonic nitrospira spp.selectively enriched from activated sludge[J].Environmental Science&Technology,2017,51(5):2720-2728.
[30]Spieck E,Hartwig C,Mc Cormack I,et al.Selective enrichment and molecular characterization of a previously uncultured Nitrospira-like bacterium from activated sludge[J].Environmental Microbiology,2006,8(3):405-415.
[31]Lücker S,Wagner M,Maixner F,et al.A Nitrospira metagenome illuminates the physiology and evolution of globally important nitrite-oxidizing bacteria[J].Proceedings of the National Academy of Sciences of the United States of America,2010,107(30):13479-13484.
[2]张自杰,林荣忱,金儒霖.排水工程下册[M].(第四版).北京:中国建筑工业出版社,2000.
[3]Arnaldos M,Pagilla K R.Implementation of a demand-side approach to reduce aeration requirements of activated sludge systems:directed acclimation of biomass and its effect at the process level[J].Water Research,2014,62:147-155.
[4]Metcalf&Eddy.Wastewater engineering:treatment and reuse(4th ed.)[M].Boston:Mc Graw-Hill,2003.
[5]Abbassi B,Dullstein S,Rbiger N.Minimization of excess sludge production by increase of oxygen concentration in activated sludge flocs;Experimental and theoretical approach[J].Water Research,2000,34(1):139-146.
[6]Liu G Q,Wang J M.Long-term low do enriches and shifts nitrifier community in activated sludge[J].Environmental Science&Technology,2013,47(10):5109-5117.
[7]Fitzgerald C M,Camejo P,Oshlag J Z,et al.Ammoniaoxidizing microbial communities in reactors with efficient nitrification at low-dissolved oxygen[J].Water Research,2015,70:38-51.
[8]Park H D,Noguera D R.Evaluating the effect of dissolved oxygen on ammonia-oxidizing bacterial communities in activated sludge[J].Water Research,2004,38(14-15):3275-3286.
[9]Arnaldos M,Kunkel S A,Stark B C,et al.Characterization of heme protein expressed by ammonia-oxidizing bacteria under low dissolved oxygen conditions[J].Applied Microbiology and Biotechnology,2014,98(7):3231-3239.
[10]Bellucci M,Ofiteru I D,Graham D W,et al.Low-dissolvedoxygen nitrifying systems exploit ammonia-oxidizing bacteria with unusually high yields[J].Applied and Environmental Microbiology,2011,77(21):7787-7796.
[11]Wagner M,Loy A.Bacterial community composition and function in sewage treatment systems[J].Current Opinion in Biotechnology,2002,13(3):218-227.
[12]Ju F,Zhang T.Bacterial assembly and temporal dynamics in activated sludge of a full-scale municipal wastewater treatment plant[J].The ISME Journal,2015,9(3):683-695.
[13]Girvan M S,Campbell C D,Killham K,et al.Bacterial diversity promotes community stability and functional resilience after perturbation[J].Environmental Microbiology,2005,7(3):301-313.
[14]Briones A,Raskin L.Diversity and dynamics of microbial communities in engineered environments and their implications for process stability[J].Current Opinion in Biotechnology,2003,14(3):270-276.
[15]Wittebolle L,Marzorati M,Clement L,et al.Initial community evenness favours functionality under selective stress[J].Nature,2009,458(7238):623-626.
[16]van de Graaf A A,de Bruijn P,Robertson L A,et al.Autotrophic growth of anaerobic ammonium-oxidizing microorganisms in a fluidized bed reactor[J].Microbiology,1996,142(8):2187-2196.
[17]国家环境保护总局.水和废水监测分析方法[M].(第四版).北京:中国环境科学出版社,2002.
[18]Liu W R,Yang D H,Chen W J,et al.High-throughput sequencing-based microbial characterization of size fractionated biomass in an anoxic anammox reactor for low-strength wastewater at low temperatures[J].Bioresource Technology,2017,231:45-52.
[19]Manser R,Gujer W,Siegrist H.Consequences of mass transfer effects on the kinetics of nitrifiers[J].Water Research,2005,39(19):4633-4642.
[20]Arnaldos M,Amerlinck Y,Rehman U,et al.From the affinity constant to the half-saturation index:understanding conventional modeling concepts in novel wastewater treatment processes[J].Water Research,2015,70:458-470.
[21]王中玮,彭永臻,王淑莹,等.不同运行方式下低溶解氧污泥微膨胀的可行性研究[J].环境科学,2011,32(8):2347-2352.Wang Z W,Peng Y Z,Wang S Y,et al.Feasibility study for limited filamentous bulking under low dissolved oxygen at different operation regimes[J].Environmental Science,2011,32(8):2347-2352.
[22]Ma J X,Wang Z W,Yang Y,et al.Correlating microbial community structure and composition with aeration intensity in submerged membrane bioreactors by 454 high-throughput pyrosequencing[J].Water Research,2013,47(2):859-869.
[23]Gao D W,Fu Y,Tao Y,et al.Linking microbial community structure to membrane biofouling associated with varying dissolved oxygen concentrations[J].Bioresource Technology,2011,102(10):5626-5633.
[24]Marzorati M,Wittebolle L,Boon N,et al.How to get more out of molecular fingerprints:practical tools for microbial ecology[J].Environmental Microbiology,2008,10(6):1571-1581.
[25]Dytczak M A,Londry K L,Oleszkiewicz J A.Activated sludge operational regime has significant impact on the type of nitrifying community and its nitrification rates[J].Water Research,2008,42(8-9):2320-2328.
[26]Bollmann A,Br-Gilissen M J,Laanbroek H J.Growth at low ammonium concentrations and starvation response as potential factors involved in niche differentiation among ammonia-oxidizing bacteria[J].Applied and Environmental Microbiology,2002,68(10):4751-4757.
[27]Bollmann A,Laanbroek H J.Continuous culture enrichments of ammonia-oxidizing bacteria at low ammonium concentrations[J].FEMS Microbiology Ecology,2001,37(3):211-221.
[28]Nowka B,Daims H,Spieck E.Comparison of oxidation kinetics of nitrite-oxidizing bacteria:nitrite availability as a key factor in niche differentiation[J].Applied and Environmental Microbiology,2015,81(2):745-753.
[29]Park M R,Park H,Chandran K.Molecular and kinetic characterization of Planktonic nitrospira spp.selectively enriched from activated sludge[J].Environmental Science&Technology,2017,51(5):2720-2728.
[30]Spieck E,Hartwig C,Mc Cormack I,et al.Selective enrichment and molecular characterization of a previously uncultured Nitrospira-like bacterium from activated sludge[J].Environmental Microbiology,2006,8(3):405-415.
[31]Lücker S,Wagner M,Maixner F,et al.A Nitrospira metagenome illuminates the physiology and evolution of globally important nitrite-oxidizing bacteria[J].Proceedings of the National Academy of Sciences of the United States of America,2010,107(30):13479-13484.