硫酸盐还原反应器污泥驯化过程中微生物群落变化分析
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摘要
采用PCR-DGGE技术,对硫酸盐还原反应器UASB污泥驯化过程中微生物群落的变化进行了分析.结果表明,污泥驯化过程中,微生物群落生物多样性与反应器硫酸盐及COD去除率呈明显的正相关,微生物群落Shannon指数大于3.45时,反应器硫酸盐去除率稳定在95%左右;对DGGE图谱中优势条带进行回收克隆并测序表明,污泥中微生物群落主要包含Firmicutes、Proteobacteria、Deinococcus-Thermus和Chloroflexi这4大类群,分别占总数的50.0%、28.6%、14.3%和7.1%;其中厌氧发酵细菌Clostridium sp.在驯化全过程中均占优势,但优势菌群的种类发生变化;厌氧细菌Chloroflexi sp.、Geopsychrobacter sp.等在污泥驯化过程中曾成为优势菌群但之后消亡;厌氧细菌Geobacter sp.则在污泥驯化过程中逐渐成为优势菌群;Desulfovibrio sp.在污泥驯化的最后2个阶段成为优势菌.
The variations of microbial community in the sludge of sulfate-reducing UASB during domestication period were analyzed by PCR-DGGE technique. The results showed that the diversity of microbial community was strongly related to the sulfate reduction and COD removal performance. The sulfate reduction rate of the reactor was about 95% when the Shannon index of microbial community was higher than 3. 45. The preponderant bands in DGGE figure were excised and cloned,and the sequencing analysis indicated there were Firmicutes,Proteobacteria,Deinococcus-Thermus and Chloroflexi in the sludge,which accounted for 50. 0%,28. 6%,14. 3%and 7. 1% of the total sequences of samples,respectively. The anaerobic fermentative bacteria of Clostridium sp. were predominant in the whole domestication period,but the predominant species was changing. Some anaerobic bacteria like Chloroflexi sp. and Geopsychrobacter sp. were detected to be dominant species,which then disappeared along with further domestication,but anaerobic bacteria Geobacter sp. became gradually predominant in the domestication process. Species of Desulfovibrio sp. were detected to be predominant only in the last two phases of domestication.
The variations of microbial community in the sludge of sulfate-reducing UASB during domestication period were analyzed by PCR-DGGE technique. The results showed that the diversity of microbial community was strongly related to the sulfate reduction and COD removal performance. The sulfate reduction rate of the reactor was about 95% when the Shannon index of microbial community was higher than 3. 45. The preponderant bands in DGGE figure were excised and cloned,and the sequencing analysis indicated there were Firmicutes,Proteobacteria,Deinococcus-Thermus and Chloroflexi in the sludge,which accounted for 50. 0%,28. 6%,14. 3%and 7. 1% of the total sequences of samples,respectively. The anaerobic fermentative bacteria of Clostridium sp. were predominant in the whole domestication period,but the predominant species was changing. Some anaerobic bacteria like Chloroflexi sp. and Geopsychrobacter sp. were detected to be dominant species,which then disappeared along with further domestication,but anaerobic bacteria Geobacter sp. became gradually predominant in the domestication process. Species of Desulfovibrio sp. were detected to be predominant only in the last two phases of domestication.
引文
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[34]Vandieken V,Mussmann M,Niemann H et al.Desulfuromonas svalbardensis sp.nov.and Desulfuromusa ferrireducens sp.nov.,psychrophilic,Fe(Ⅲ)-reducing bacteria isolated from Arctic sediments,Svalbard[J].International Journal of Systematic and Evolutionary Microbiology,2006,56(5):1133-1139.
[35]Widdel F,Hansen T A.The dissimilatory sulfate-and sulfurreducing bacteria[A].In:The Procaryotes[M].(2nd ed.).New York:Springer,1992.583-624.
[36]Oude Elferink S J W H,Vorstman W J C,Sopjes A,et al.Characterization of the sulfate-reducing and syntrophic population in granular sludge from a full-scale anaerobic reactor treating papermill wastewater[J].FEMS Microbiology Ecology,1998,27(2):185-194.
[2]Liu W T,Chan O C,Fang H H P.Microbial community dynamics during start-up of acidogenic anaerobic reactors[J].Water Research,2002,36(13):3203-3210.
[3]赵丹,任南琪,王爱杰,等.产酸相稳定发酵类型微生物生态学研究[J].环境科学与技术,2003,26(6):37-38,48.
[4]张斌,孙宝盛,刘慧娜,等.处理不同废水MBR系统中微生物群落结构的比较[J].环境科学,2008,29(10):2944-2949.
[5]任南琪,王爱杰.产酸脱硫反应器中碳硫比对群落生态特征的影响[J].环境科学,2002,23(4):52-56.
[6]王爱杰,任南琪,黄志,等.产酸脱硫反应器中COD/SO2-4比制约的群落生态演替规律[J].环境科学,2002,23(2):34-38.
[7]苏俊峰,马放,王弘宇,等.利用PCR-DGGE技术分析生物陶粒硝化反应器中微生物群落动态[J].环境科学学报,2007,27(3):386-390.
[8]Ding Y,Wu W X,Han Z Y,et al.Correlation of reactor performance and bacterial community composition during the removal of trimethylamine in three-stage biofilters[J].Biochemical Engineering Journal,2008,38(2):248-258.
[9]Miura Y,Hiraiwa M N,Ito T,et al.Bacterial community structures in MBRs treating municipal wastewater:Relationship between community stability and reactor performance[J].Water Research,2007,41(3):627-637.
[10]任随周,郭俊,曾国驱,等.处理印染废水的厌氧折流板反应器中的微生物种群组成及分布规律[J].生态学报,2005,25(9):2297-2303.
[11]刘广民,任南琪,王爱杰,等.产酸-硫酸盐还原系统中产酸菌的发酵类型及其与SRB的协同作用[J].环境科学学报,2004,24(5):782-788.
[12]Colleran E,Pender S.Mesophilic and thermophilic anaerobic digestion of sulphate-containing wastewaters[J].Water Science and Technology,2002,45(10):231-235.
[13]Sanz J L,Kochling T.Molecular biology techniques used in wastewater treatment:An overview[J].Process Biochemistry,2007,42(2):119-133.
[14]Muyzer G,de Waal E C,Uitterlinden A G.Profiling of complex microbial population by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes encoding for16S rRNA[J].Applied and Environmental Microbiology,1993,59(3):695-700.
[15]刘新春,吴成强,张昱,等.PCR-DGGE法用于活性污泥系统中微生物群落结构变化的解析[J].生态学报,2005,25(4):842-847.
[16]李建军,廖东奇,许玫英,等.生物滴滤池对BTEX的去除及相应细菌群落分析[J].环境科学,2013,34(7):2552-2559.
[17]欧阳科,刘俊新.膜生物反应器与传统活性污泥反应器内生物群落特征[J].环境科学,2009,30(2):499-503.
[18]罗海峰,齐鸿雁,薛凯,等.PCR-DGGE技术在农田土壤微生物多样性研究中的应用[J].生态学报,2003,23(8):1570-1575.
[19]Aburto-Medina A,Adetutu E M,Aleer S,et al.Comparison of indigenous and exogenous microbial populations during slurry phase biodegradation of long-term hydrocarbon-contaminated soil[J].Biodegradation,2012,23(6):813-822.
[20]刘敏,朱开玲,李洪波,等.应用PCR-DGGE技术分析黄海冷水团海域的细菌群落组成[J].环境科学,2008,29(4):1082-1091.
[21]APHA.Standard methods for the examination of water and wastewater[M].(20th ed.).USA Washington DC:American Public Health Association,1998.
[22]Zhou J Z,Bruns M A,Tiedje J M.DNA recovery from soils of diverse composition[J].Applied and Environmental Microbiology,1996,62(2):316-322.
[23]郑小红,肖琳,任晶,等.玄武湖微囊藻水华暴发及衰退期细菌群落变化分析[J].环境科学,2008,29(10):2956-2962.
[24]王爱杰,任南琪,刘伟,等.产酸脱硫反应器中SRB种群的功能与地位[J].中国环境科学,2001,21(2):119-123.
[25]Matthies C,Kuhner C H,Acker G,et al.Clostridium uliginosum sp.nov.,a novel acid-tolerant,anaerobic bacterium with connecting filaments[J].International Journal of Systematic and Evolutionary Microbiology,2001,51(3):1119-1125.
[26]Kaksonen A H,Plumb J J,Robertson W J,et al.Culturable diversity and community fatty acid profiling of sulfate-reducing fluidized-bed reactors treating acidic,metal-containing wastewater[J].Geomicrobiology Journal,2004,21(7):469-480.
[27]任南琪,赵阳国,王爱杰,等.PCR-SSCP技术分析碱度影响下硫酸盐还原反应器中微生物群落动态[J].中国科学(C辑):生命科学,2006,36(1):51-58.
[28]Vladár P,Rusznyák A,Márialigeti K,et al.Diversity of sulfatereducing bacteria inhabiting the rhizosphere of phragmites australis in Lake Velencei(Hungary)revealed by a combined cultivation-based and molecular approach[J].Microbial Ecology,2008,56(1):64-75.
[29]Mallet C,Basset M,Fonty G,et al.Microbial population dynamics in the sediments of a eutrophic lake(Aydat,France)and characterization of some heterotrophic bacterial isolates[J].Microbial Ecology,2004,48(1):66-77.
[30]Bjornsson 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]Roest K,Heilig H G H J,Smidt H,et al.Community analysis of a full-scale anaerobic bioreactor treating paper mill wastewater[J].Systematic and Applied Microbiology,2005,28(2):175-185.
[32]Cord-Ruwisch R,Lovley D R,Schink B.Growth of Geobacter sulfurreducens with acetate in syntrophic cooperation with hydrogen-oxidizing anaerobic partners[J].Applied and Environmental Microbiology,1998,64(6):2232-2236.
[33]Ramos D T,da Silva M L B,Chiaranda H S,et al.Biostimulation of anaerobic BTEX biodegradation under fermentative methanogenic conditions at source-zone groundwater contaminated with a biodiesel blend(B20)[J].Biodegradation,2013,24(3):333-341.
[34]Vandieken V,Mussmann M,Niemann H et al.Desulfuromonas svalbardensis sp.nov.and Desulfuromusa ferrireducens sp.nov.,psychrophilic,Fe(Ⅲ)-reducing bacteria isolated from Arctic sediments,Svalbard[J].International Journal of Systematic and Evolutionary Microbiology,2006,56(5):1133-1139.
[35]Widdel F,Hansen T A.The dissimilatory sulfate-and sulfurreducing bacteria[A].In:The Procaryotes[M].(2nd ed.).New York:Springer,1992.583-624.
[36]Oude Elferink S J W H,Vorstman W J C,Sopjes A,et al.Characterization of the sulfate-reducing and syntrophic population in granular sludge from a full-scale anaerobic reactor treating papermill wastewater[J].FEMS Microbiology Ecology,1998,27(2):185-194.