CANON工艺处理猪场沼液的启动及微生物种群结构分析
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摘要
在连续流合建式反应器内接种普通活性污泥,以实际猪场沼液作为进水,保持温度为(30±1)℃,控制低溶解氧(DO浓度为(0.5±0.1)mg·L~(-1)),通过分阶段提高氨氮浓度的方式启动CANON(Completely autotrophic nitrogen removal over nitrite)工艺,同时采用高通量测序技术对反应器启动前后的微生物种群结构进行分析.结果表明,反应器可在210 d实现成功启动CANON工艺,启动成功后,当猪场沼液进水氨氮浓度为450 mg·L~(-1)时,氨氮和总氮的去除率分别达到69.8%和61.1%;出水(NO_3~--N+NO_2~--N)/ΔNH_4~+接近0.11.同时,CANON工艺启动成功后,微生物种群结构发生了很大的变化:其优势菌门为变形菌门(Proteobacteria)(36.16%)、绿弯菌门(Chloroflexi)(14.22%)、浮霉菌门(Planctomycetes)(11.07%)、酸杆菌门(Acidobacteria)(10.41%)和绿菌门(Chlorobi)(12.82%);与脱氮功能相关的菌属为Candidatus_Brocadia (ANAMMOX菌)、Nitrosomonas(AOB菌)、Comamonadaceae(反硝化菌)和Xanthomonadaceae(反硝化菌)等.
A successful start-up of a completely autotrophic nitrogen removal over nitrite( CANON) process treating anaerobic digester liquor of swine wastewater( ADLSW) was achieved in a continuous-flow biofilm reactor( CFBR) by gradually increasing the influent ammonium( NH_4~+-N) concentration and controlling the low dissolve oxygen( DO)(( 0. 5 ± 0. 1) mg·L~(-1)). The temperature was maintained at(30± 1) ℃. High-throughput sequencing technique was used to monitor the evolution of the bacterial community. The results demonstrate that a successful start-up of CANON process was accomplished in 210 d. When the influent ammonia nitrogen concentration was 450 mg·L~(-1),the average ammonium removal efficiency and total nitrogen removal efficiency was up to 69.8% and 61.1%,respectively. And the effluent( NO_3~--N+NO_2~--N)/ΔNH_4~+-N ratio was closed to 0.11. At the same time,compared to the seed sludge,the microbial communities had remarkable changes in the sludge of CANON process. The dominant microbial population at phylum level wereProteobacteria( 36.16%),Chloroflexi( 14.22%),Planctomycetes( 11. 07%),Acidobacteria( 10. 41%) and Chlorobi( 12. 82%).The dominant bacterial species related to nitrogen transformation were Candidatus_Brocadia,Nitrosomonas,Comamonadaceae and Xanthomonadaceae.
A successful start-up of a completely autotrophic nitrogen removal over nitrite( CANON) process treating anaerobic digester liquor of swine wastewater( ADLSW) was achieved in a continuous-flow biofilm reactor( CFBR) by gradually increasing the influent ammonium( NH_4~+-N) concentration and controlling the low dissolve oxygen( DO)(( 0. 5 ± 0. 1) mg·L~(-1)). The temperature was maintained at(30± 1) ℃. High-throughput sequencing technique was used to monitor the evolution of the bacterial community. The results demonstrate that a successful start-up of CANON process was accomplished in 210 d. When the influent ammonia nitrogen concentration was 450 mg·L~(-1),the average ammonium removal efficiency and total nitrogen removal efficiency was up to 69.8% and 61.1%,respectively. And the effluent( NO_3~--N+NO_2~--N)/ΔNH_4~+-N ratio was closed to 0.11. At the same time,compared to the seed sludge,the microbial communities had remarkable changes in the sludge of CANON process. The dominant microbial population at phylum level wereProteobacteria( 36.16%),Chloroflexi( 14.22%),Planctomycetes( 11. 07%),Acidobacteria( 10. 41%) and Chlorobi( 12. 82%).The dominant bacterial species related to nitrogen transformation were Candidatus_Brocadia,Nitrosomonas,Comamonadaceae and Xanthomonadaceae.
引文
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包鹏,王淑莹,高瑶远,等.2017.利用高通量测序技术分析不同溶解氧条件下硝化活性污泥的菌群结构特性[J].北京工业大学学报,43(5):801-808
Cardinali-Rezende J,Pereira Z L,Sanz J L,et al.2012.Bacterial and archaeal phylogenetic diversity associated with swine sludge from an anaerobic treatment lagoon[J].World Journal of Microbiology and Biotechnology,28(11):3187-3195
初里冰,张兴文,李晓惠,等.2005.一种新型生物脱氮工艺-CANON工艺[J].齐鲁工业大学学报,19(1):21-24
Daims H,Nielsen J L,Nielsen P H,et al.2001.In situ characterization of nitrospira-like nitrite-oxidizing bacteria active in wastewater treatment plants[J].Appl Environ Microbiol,67(11):5273-5284
Daverey A,Su S H,Huang Y T,et al.2012.Nitrogen removal from optoelectronic wastewater using the simultaneous partial nitrification,anaerobic ammonium oxidation and denitrification(SNAD)in sequencing batch reactor[J].Bioresour Technology,113:225-231
Foesel B U,Rohde M,Overmann J,et al.2013.Blastocatella fastidiosa gen.nov.,sp.nov.,isolated from semiari savanna soil-The first described species of Acidobacteria subdivision 4[J].Systematic and Applied Microbiology,36(2):82-89
付昆明,左早荣,仇付国.2014.陶粒CANON反应器的接种启动与运行[J].环境科学学报,35(3):995-1000
Fuerst J A.2010.Beyond prokaryotes and eukaryotes:Planctomycetes and cell organization[J].Nature Education,3(9):44
龚维红,陈玮.2005.城市污泥好氧发酵菌种放线菌的初步筛选[J].生物技术,15(2):41-43
宫正,刘思彤,杨凤林,等.2008.启动炭管膜曝气生物膜反应器实现全程自养脱氮[J].环境科学,29(5):1221-1226
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国家环境保护总局.2002.水和废水监测分析方法(第四版)[M].北京:中国环境科学出版社.258-280
韩伟铖,颜成,周立祥.2017.规模化猪场废水常规生化处理的效果及原因剖析[J].农业环境科学学报,36(5):989-995
Hao X,Heijnen J J,VanLoosdrecht M C.2002.Model-base devaluation of temperature and inflow variations on a partial nitrificationANAMMOX biofilm process[J].Water Research,36(19):4839
Hu B L,Zheng P,Tang C J,et al.2010.Identification and quantification of anammox bacteria in eight nitrogenremoval reactors[J].Water Research,44(17):5014-5020
Hulle S W V,Volcke E I,Teruel J L,et al.2007.Influence oftemperature and p H on the kinetics of the Sharon nitritation process[J].Journal of Chemical Technology&Biotechnology,82(5):471-480
Jing X,Dong L,Hai Y,et al.2014.Performance and microbial community of completely autotrophic nitrogen removal over nitrite(CANON)process in two membrane bioreactors(MBR)fed with different substrate levels[J].Bioresource Technology,152:185-191
李东,何永平,张肖静,等.2014.MBR系统CANON工艺的快速启动及微生物种群特征[J].中国环境科学,34(11):2788-2795
刘彩霞,董玉红,焦如珍.2016.森林土壤中酸杆菌门多样性研究进展[J].世界林业研究,29(6):17-22
Liu L,You Q,Gibson V,et al.2015.Treatment of swine wastewater in aerobic granular reactors:Comparison of different seed granules as factors[J].Frontiers of Environmental Science&Engineering,9(6):1139-1148
Liu S,Yang F,Gong Z,et al.2008.Assessment of the positive effect of salinity on the nitrogen removal performance and microbial composition during the start-up of CANON process[J].Applied Microbiology and Biotechnology,80(2):339-348
Liu T,Dong L,Jie Z,et al.2011.Phylogenetic and microbial community analysis based on amo A gene and 16S r DNA innitrosification biofilm reactor[J].Environmental Biotechnologyand Materials Engineering,183:1051-1056
Liu T,Li D,Zeng H P,et al.2012.Biodiversity and quantification of functional bacteria in completely au.totrophic nitrogen-removal over nitrite(CANON)process[J].Bioresource Technology,118:399-406
刘竹寒,岳秀,于广平,等.2017.CANON在SBAF中的快速启动及其微生物特征[J].环境科学学报,38(1):253-259
Lotti T,Star W,Kleerebezem R,et al.2012.The effect of nitrite inhibition on the anammox process[J].Water Research,46(8):2559-2569
鲁航,信欣,管蕾,等.2016.部分半硝化AGS-SBR工艺的启动及其种群结构分析[J].环境科学学报,37(12):4741-4749
Muyzer G,Dewaal E C,Uitterlinden A G.1993.Profiling of complex microbial-populations by denaturing gradient gel-electrophoresis analysis of polymerase chain reaction-amplified genes-coding for 16s Ribosomal-Rna[J].Applied and Environmental Microbiology,59(3):695-700
Peng L,Wei X,Zuo J,et al.2013.Hydrogenotrophic denitrification for tertiary nitrogen removal from municipal wastewater using membrane diffusion packed-bed bioreactor[J].Bioresource Technology,144(5):452-459
Prakash O,Green S J,Jasrotia P,et al.2012.Rhodanobacter denitrificans sp.nov.,isolated from nitrate-rich zones of a contaminated aquifer[J].International Journal of Systematic and Evolutionary Microbiology,62(10):2457-2462
Sadaie T,Sadaie A,Takada M,et al.2007.Reducing sludge production and the domination of Comamonadaceae by reducing the oxygen supply in the wastewater treatment procedure of a food-processing factory[J].Agricultural and Biological Chemistry,71:791-799
Sliekers A O,Derwort N,Campos-Gomez J L,et al.2002.Completely autotrophic nitrogen removal over nitrite in one single reactor[J].Water Research,36(10):2475-2482
Sliekers A O,Third K A,Abma W,et al.2003.CANON and Anammox in a gas lift reactor[J].FEMS Microbiology Letters,210:339-344
孙洪伟,吕心涛,魏雪芬,等.2016.游离氨(FA)耦合曝气时间对硝化菌活性的抑制影响[J].环境科学,37(3):1075-1081
孙延芳,韩晓宇,张树军,等.2017.颗粒+絮体污泥CANON工艺的启动与SRT影响研究[J].环境科学学报,38(2):672-678
孙智毅,2017.一株异养硝化-好氧反硝化Cupriavidus sp.S1的筛选及降解特性研究[D].太原:太原理工大学
王明月.2017.猪场消毒的重要性及注意事项[J].现代畜牧科技,32(8):9-10
王衫允.2016.低氨氮浓度厌氧氨氧化工艺强化及颗粒污泥菌群特性研究[D].哈尔滨:哈尔滨工业大学
Ward N L,Challacombe J F,Janssen P H,et al.2009.Three genomes from the phylum Acidobacteria provide insight into the lifestyles of these microorganisms in soils[J].Applied and Environmental Microbiology,75(7):2046-2056
Weissbrodt D G,Shani N,Holliger C,et al.2014.Linking bacterial population dynamics and nutrient removal in the granular sludge biofilm ecosystem engineered for wastewater treatment[J].FEMSMicrobiology Ecology,88(3):579-595
吴斌,王必尊,王丽霞,等.2018.不同催花肥对莲雾根际土壤细菌群落多样性的影响[J].环境科学研究,31(4):732-741
Xiu Y,Guang P Y,Yu Q L,et al.2018.Effect of dissolved oxygen on nitrogen removal and the microbial community of the completely autotrophic nitrogen removal over nitrite process in a submerged aerated biological filter[J].Bioresource Technology,254:67-74
闫媛,黎力,王亚宜,等.2015.采用高通量测序分析全程自养脱氮(CANON)系统不同脱氮效能下的微生物群落结构[J].北京工业大学学报,41(10):1485-1492
闫园园,李子富,程世昆,等.2013.养殖场厌氧发酵沼液处理研究进展[J].中国沼气,31(5):48-52
Yang Z H,Xiao Y,Zeng G M,et al.2007.Comparison of methods for total community DNA extraction and purification from compost[J].Applied Microbiology and Biotechnology,74(4):918-925
Yoon D N,Park S J,Kim S J,et al.2010.Isolation,characterization,and abundance of filamentous members of caldilineae in activated sludge[J].Journal of Microbiology,48(3):275-283
Zhang H,Sekiguchi Y,Hanada S,et al.2003.Gemmatimonas aurantiaca gen.nov.,sp.nov.,a Gram-negative,aerobic,polyphosphateaccumulating micro-organism,the first cultured representative of the new bacterial phylum Gemmatimonadetes phyl.nov.[J].International Journal of Systematic and Evolutionary Microbiology,53(4):1155-1163
张杰,成朔,李冬,等.2018.AUSB中置曝气启动连续流全程自养脱氮工艺[J].哈尔滨工业大学学报,50(2):1-7
张克强,高怀友.2004.畜禽养殖业污染物处理与处置[M].北京:化学工业出版社.26-30
Zhang T,Shao M F,Ye L.2012.454 Pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants[J].The ISME Journal,6(6):1137-1147
Zhang X J,Li D,Liang Y H,et al.2013.Application of membrane bioreactor for completely autotrophic nitrogen removal over nitrite(CANON)process[J].Chemosphere,93(11):2832-2838
张肖静,李冬,马静,等.2016.SNAD生物膜厌氧氨氧化活性的氨氮抑制动力学研究[J].中国环境科学,36(10):2957-2963
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左早荣,付昆明,仇付国,等.2013.CANON工艺的研究现状及面临困难分析[J].水处理技术,39(9):15-19
Anthonisen A C,Loehr R C,Prakasam T B S,et al.1976.Inhibition of nitrification by ammonia and nitrous acid[J].Journal of Water Pollution Control Federation,48(5):835-852
包鹏,王淑莹,高瑶远,等.2017.利用高通量测序技术分析不同溶解氧条件下硝化活性污泥的菌群结构特性[J].北京工业大学学报,43(5):801-808
Cardinali-Rezende J,Pereira Z L,Sanz J L,et al.2012.Bacterial and archaeal phylogenetic diversity associated with swine sludge from an anaerobic treatment lagoon[J].World Journal of Microbiology and Biotechnology,28(11):3187-3195
初里冰,张兴文,李晓惠,等.2005.一种新型生物脱氮工艺-CANON工艺[J].齐鲁工业大学学报,19(1):21-24
Daims H,Nielsen J L,Nielsen P H,et al.2001.In situ characterization of nitrospira-like nitrite-oxidizing bacteria active in wastewater treatment plants[J].Appl Environ Microbiol,67(11):5273-5284
Daverey A,Su S H,Huang Y T,et al.2012.Nitrogen removal from optoelectronic wastewater using the simultaneous partial nitrification,anaerobic ammonium oxidation and denitrification(SNAD)in sequencing batch reactor[J].Bioresour Technology,113:225-231
Foesel B U,Rohde M,Overmann J,et al.2013.Blastocatella fastidiosa gen.nov.,sp.nov.,isolated from semiari savanna soil-The first described species of Acidobacteria subdivision 4[J].Systematic and Applied Microbiology,36(2):82-89
付昆明,左早荣,仇付国.2014.陶粒CANON反应器的接种启动与运行[J].环境科学学报,35(3):995-1000
Fuerst J A.2010.Beyond prokaryotes and eukaryotes:Planctomycetes and cell organization[J].Nature Education,3(9):44
龚维红,陈玮.2005.城市污泥好氧发酵菌种放线菌的初步筛选[J].生物技术,15(2):41-43
宫正,刘思彤,杨凤林,等.2008.启动炭管膜曝气生物膜反应器实现全程自养脱氮[J].环境科学,29(5):1221-1226
Gonzalez-Martinez A,Rodriguez-Sanchez A,Garcia-Ruiz M J,et al.2016.Performance and bacterial community dynamics of a CANONbioreactor acclimated from high to low operational temperatures[J].Chemical Engineering Journal,287:557-567
国家环境保护总局.2002.水和废水监测分析方法(第四版)[M].北京:中国环境科学出版社.258-280
韩伟铖,颜成,周立祥.2017.规模化猪场废水常规生化处理的效果及原因剖析[J].农业环境科学学报,36(5):989-995
Hao X,Heijnen J J,VanLoosdrecht M C.2002.Model-base devaluation of temperature and inflow variations on a partial nitrificationANAMMOX biofilm process[J].Water Research,36(19):4839
Hu B L,Zheng P,Tang C J,et al.2010.Identification and quantification of anammox bacteria in eight nitrogenremoval reactors[J].Water Research,44(17):5014-5020
Hulle S W V,Volcke E I,Teruel J L,et al.2007.Influence oftemperature and p H on the kinetics of the Sharon nitritation process[J].Journal of Chemical Technology&Biotechnology,82(5):471-480
Jing X,Dong L,Hai Y,et al.2014.Performance and microbial community of completely autotrophic nitrogen removal over nitrite(CANON)process in two membrane bioreactors(MBR)fed with different substrate levels[J].Bioresource Technology,152:185-191
李东,何永平,张肖静,等.2014.MBR系统CANON工艺的快速启动及微生物种群特征[J].中国环境科学,34(11):2788-2795
刘彩霞,董玉红,焦如珍.2016.森林土壤中酸杆菌门多样性研究进展[J].世界林业研究,29(6):17-22
Liu L,You Q,Gibson V,et al.2015.Treatment of swine wastewater in aerobic granular reactors:Comparison of different seed granules as factors[J].Frontiers of Environmental Science&Engineering,9(6):1139-1148
Liu S,Yang F,Gong Z,et al.2008.Assessment of the positive effect of salinity on the nitrogen removal performance and microbial composition during the start-up of CANON process[J].Applied Microbiology and Biotechnology,80(2):339-348
Liu T,Dong L,Jie Z,et al.2011.Phylogenetic and microbial community analysis based on amo A gene and 16S r DNA innitrosification biofilm reactor[J].Environmental Biotechnologyand Materials Engineering,183:1051-1056
Liu T,Li D,Zeng H P,et al.2012.Biodiversity and quantification of functional bacteria in completely au.totrophic nitrogen-removal over nitrite(CANON)process[J].Bioresource Technology,118:399-406
刘竹寒,岳秀,于广平,等.2017.CANON在SBAF中的快速启动及其微生物特征[J].环境科学学报,38(1):253-259
Lotti T,Star W,Kleerebezem R,et al.2012.The effect of nitrite inhibition on the anammox process[J].Water Research,46(8):2559-2569
鲁航,信欣,管蕾,等.2016.部分半硝化AGS-SBR工艺的启动及其种群结构分析[J].环境科学学报,37(12):4741-4749
Muyzer G,Dewaal E C,Uitterlinden A G.1993.Profiling of complex microbial-populations by denaturing gradient gel-electrophoresis analysis of polymerase chain reaction-amplified genes-coding for 16s Ribosomal-Rna[J].Applied and Environmental Microbiology,59(3):695-700
Peng L,Wei X,Zuo J,et al.2013.Hydrogenotrophic denitrification for tertiary nitrogen removal from municipal wastewater using membrane diffusion packed-bed bioreactor[J].Bioresource Technology,144(5):452-459
Prakash O,Green S J,Jasrotia P,et al.2012.Rhodanobacter denitrificans sp.nov.,isolated from nitrate-rich zones of a contaminated aquifer[J].International Journal of Systematic and Evolutionary Microbiology,62(10):2457-2462
Sadaie T,Sadaie A,Takada M,et al.2007.Reducing sludge production and the domination of Comamonadaceae by reducing the oxygen supply in the wastewater treatment procedure of a food-processing factory[J].Agricultural and Biological Chemistry,71:791-799
Sliekers A O,Derwort N,Campos-Gomez J L,et al.2002.Completely autotrophic nitrogen removal over nitrite in one single reactor[J].Water Research,36(10):2475-2482
Sliekers A O,Third K A,Abma W,et al.2003.CANON and Anammox in a gas lift reactor[J].FEMS Microbiology Letters,210:339-344
孙洪伟,吕心涛,魏雪芬,等.2016.游离氨(FA)耦合曝气时间对硝化菌活性的抑制影响[J].环境科学,37(3):1075-1081
孙延芳,韩晓宇,张树军,等.2017.颗粒+絮体污泥CANON工艺的启动与SRT影响研究[J].环境科学学报,38(2):672-678
孙智毅,2017.一株异养硝化-好氧反硝化Cupriavidus sp.S1的筛选及降解特性研究[D].太原:太原理工大学
王明月.2017.猪场消毒的重要性及注意事项[J].现代畜牧科技,32(8):9-10
王衫允.2016.低氨氮浓度厌氧氨氧化工艺强化及颗粒污泥菌群特性研究[D].哈尔滨:哈尔滨工业大学
Ward N L,Challacombe J F,Janssen P H,et al.2009.Three genomes from the phylum Acidobacteria provide insight into the lifestyles of these microorganisms in soils[J].Applied and Environmental Microbiology,75(7):2046-2056
Weissbrodt D G,Shani N,Holliger C,et al.2014.Linking bacterial population dynamics and nutrient removal in the granular sludge biofilm ecosystem engineered for wastewater treatment[J].FEMSMicrobiology Ecology,88(3):579-595
吴斌,王必尊,王丽霞,等.2018.不同催花肥对莲雾根际土壤细菌群落多样性的影响[J].环境科学研究,31(4):732-741
Xiu Y,Guang P Y,Yu Q L,et al.2018.Effect of dissolved oxygen on nitrogen removal and the microbial community of the completely autotrophic nitrogen removal over nitrite process in a submerged aerated biological filter[J].Bioresource Technology,254:67-74
闫媛,黎力,王亚宜,等.2015.采用高通量测序分析全程自养脱氮(CANON)系统不同脱氮效能下的微生物群落结构[J].北京工业大学学报,41(10):1485-1492
闫园园,李子富,程世昆,等.2013.养殖场厌氧发酵沼液处理研究进展[J].中国沼气,31(5):48-52
Yang Z H,Xiao Y,Zeng G M,et al.2007.Comparison of methods for total community DNA extraction and purification from compost[J].Applied Microbiology and Biotechnology,74(4):918-925
Yoon D N,Park S J,Kim S J,et al.2010.Isolation,characterization,and abundance of filamentous members of caldilineae in activated sludge[J].Journal of Microbiology,48(3):275-283
Zhang H,Sekiguchi Y,Hanada S,et al.2003.Gemmatimonas aurantiaca gen.nov.,sp.nov.,a Gram-negative,aerobic,polyphosphateaccumulating micro-organism,the first cultured representative of the new bacterial phylum Gemmatimonadetes phyl.nov.[J].International Journal of Systematic and Evolutionary Microbiology,53(4):1155-1163
张杰,成朔,李冬,等.2018.AUSB中置曝气启动连续流全程自养脱氮工艺[J].哈尔滨工业大学学报,50(2):1-7
张克强,高怀友.2004.畜禽养殖业污染物处理与处置[M].北京:化学工业出版社.26-30
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