碳源对O/A-F/F模式积累内源聚合物及反硝化的影响
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摘要
好氧/缺氧-盛宴/饥饿(O/A-F/F)选择模式能够在好氧段实现活性污泥积累内源聚合物的同时在缺氧段原位利用内源聚合物驱动反硝化.为了深入探究不同的碳源类型对O/A-F/F模式下内源聚合物积累和内源反硝化的影响,实验以乙酸和葡萄糖为主要碳源探究内源聚合物积累和内源反硝化特性以及富集的活性污泥菌群的结构和功能.结果表明,在O/A-F/F选择模式下,当进水化学需氧量(COD)为500 mg·L~(-1)左右时,以乙酸为主要碳源系统(Ac-SBR)和以葡萄糖为主要碳源的系统(Gc-SBR)均能实现40 mg·L~(-1)的硝酸盐氮的内源去除,且各系统均实现了部分短程反硝化.但Ac-SBR实现了更高的亚硝酸盐的积累.乙酸有利于内源聚羟基脂肪酸酯(PHA)积累并驱动内源反硝化过程,PHA产率为0.52,平均反硝化速率(DNR)为9.65 mg·(L·h)-1.Gc-SBR系统能够实现PHA和糖原(Gly)的同时积累,但Gly产率高于PHA产率,分别为0.36和0.17,DNR为4.35 mg·(L·h)-1.Gly是实现内源反硝化过程的主要驱动力,反硝化脱氮贡献率占总量的77%.16S rRNA高通量测序表明Proteobacteria门中的β-Proteobacteria在Ac-SBR中为优势菌纲,菌群丰度为40.56%,而在Gc-SBR中菌群丰度为18.05%.α-Proteobacteria可能在Gc-SBR中贡献了微生物的糖原积累.β-Proteobacteria、Unclassified Bacteroidetes和Lgnavibacteria在Ac-SBR中贡献了内源PHA积累.
To accumulate endogenous polymers during the aerobic phase,the aerobic/anoxic-feast/famine(O/A-F/F) selection mode can be used.It can also be used in situ for endogenous denitrification by activated sludge during the anoxic phase.To further explore the effect of carbon sources on the activated sludge accumulation of endogenous polymers and endogenous denitrification,this study used acetic and glucose as the main carbon sources to investigate the accumulation of endogenous polymers,endogenous denitrification,and the structure and function of enriched activated sludge.The results show that acetic(Ac-SBR) and glucose(Gc-SBR) as the main carbon source systems achieved a 40 mg·L~(-1) nitrate removal by endogenous denitrification when the influent chemical oxygen demand(COD) was ~ 500 mg·L~(-1) in the O/A-F/F selection mode.Both the Ac-SBR and Gc-SBR achieved partial denitrification,but the nitrite accumulation of the Ac-SBR was higher than that of the Gc-SBR.Acetic is favorable for the accumulation of endogenous polyhydroxyalkanoate(PHA);PHA drives the endogenous denitrification.The yield of PHA was 0.52 and the denitrification rate(DNR) was 9.65 mg·(L·h)-1.The Gc-SBR system achieved the simultaneous accumulation of PHA and glycogen(Gly).The yield of Gly was higher than that of PHA and the DNR driven by Gly was 4.35 mg·(L·h)-1.The Gly was the main driving force to achieve endogenous denitrification and contributed to 77% of the total nitrogen removal.The 16 S rRNA high-throughput sequencing analysis of activated sludge flora shows that the class of β-Proteobacteria in the Proteobacteria was dominant,with an abundance of 40.56% in the Ac-SBR.However,the abundance of β-Proteobacteria was only 18.05% in the Gc-SBR.The class of α-Proteobacteria contributes to glycogen accumulation in the Gc-SBR.The PHA can be accumulated by β-Proteobacteria,Unclassified Bacteroidetes,and Lgnavibacteria in the Ac-SBR.
To accumulate endogenous polymers during the aerobic phase,the aerobic/anoxic-feast/famine(O/A-F/F) selection mode can be used.It can also be used in situ for endogenous denitrification by activated sludge during the anoxic phase.To further explore the effect of carbon sources on the activated sludge accumulation of endogenous polymers and endogenous denitrification,this study used acetic and glucose as the main carbon sources to investigate the accumulation of endogenous polymers,endogenous denitrification,and the structure and function of enriched activated sludge.The results show that acetic(Ac-SBR) and glucose(Gc-SBR) as the main carbon source systems achieved a 40 mg·L~(-1) nitrate removal by endogenous denitrification when the influent chemical oxygen demand(COD) was ~ 500 mg·L~(-1) in the O/A-F/F selection mode.Both the Ac-SBR and Gc-SBR achieved partial denitrification,but the nitrite accumulation of the Ac-SBR was higher than that of the Gc-SBR.Acetic is favorable for the accumulation of endogenous polyhydroxyalkanoate(PHA);PHA drives the endogenous denitrification.The yield of PHA was 0.52 and the denitrification rate(DNR) was 9.65 mg·(L·h)-1.The Gc-SBR system achieved the simultaneous accumulation of PHA and glycogen(Gly).The yield of Gly was higher than that of PHA and the DNR driven by Gly was 4.35 mg·(L·h)-1.The Gly was the main driving force to achieve endogenous denitrification and contributed to 77% of the total nitrogen removal.The 16 S rRNA high-throughput sequencing analysis of activated sludge flora shows that the class of β-Proteobacteria in the Proteobacteria was dominant,with an abundance of 40.56% in the Ac-SBR.However,the abundance of β-Proteobacteria was only 18.05% in the Gc-SBR.The class of α-Proteobacteria contributes to glycogen accumulation in the Gc-SBR.The PHA can be accumulated by β-Proteobacteria,Unclassified Bacteroidetes,and Lgnavibacteria in the Ac-SBR.
引文
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[16]Zeng R J,Van Loosdrecht M C M,Yuan Z G,et al.Metabolic model for glycogen-accumulating organisms in anaerobic/aerobic activated sludge systems[J].Biotechnology and Bioengineering,2003,81(1):92-105.
[17]Cui Y W,Zhang H Y,Ding J R,et al.The effects of salinity on nitrification using halophilic nitrifiers in a Sequencing Batch Reactor treating hypersaline wastewater[J].Scientific Reports,2016,6:24825.
[18]Amato K R,Yeoman C J,Kent A,et al.Habitat degradation impacts black howler monkey(Alouatta pigra)gastrointestinal microbiomes[J].ISME Journal,2013,7(7):1344-1353.
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[20]Beun J J,Paletta F,Van Loosdrecht M C M,et al.Stoichiometry and kinetics of poly-β-hydroxybutyrate metabolism in aerobic,slow growing,activated sludge cultures[J].Biotechnology and Bioengineering,2000,67(4):379-389.
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[22]Shimada T,Zilles J,Raskin L,et al.Carbohydrate storage in anaerobic sequencing batch reactors[J].Water Research,2007,41(20):4721-4729.
[23]Pittmann T,Steinmetz H.Influence of operating conditions for volatile fatty acids enrichment as a first step for polyhydroxyalkanoate production on a municipal waste water treatment plant[J].Bioresource Technology,2013,148:270-276.
[24]Du R,Cao S B,Niu M,et al.Performance of partialdenitrification process providing nitrite for anammox in sequencing batch reactor(SBR)and upflow sludge blanket(USB)reactor[J].International Biodeterioration&Biodegradation,2017,122:38-46.
[25]Cao S B,Wang S Y,Peng Y Z,et al.Achieving partial denitrification with sludge fermentation liquid as carbon source:The effect of seeding sludge[J].Bioresource Technology,2013,149:570-574.
[26]Ge S J,Peng Y Z,Wang S Y,et al.Nitrite accumulation under constant temperature in anoxic denitrification process:The effects of carbon sources and COD/NO3-N[J].Bioresource Technology,2012,114:137-143.
[27]Ince O,Basak B,Ince B K,et al.Effect of nitrogen deficiency during SBR operation on PHA storage and microbial diversity[J].Environmental Technology,2012,33(16):1827-1837.
[28]Venkateswar Reddy M,Venkata Mohan S.Effect of substrate load and nutrients concentration on the polyhydroxyalkanoates(PHA)production using mixed consortia through wastewater treatment[J].Bioresource Technology,2012,114:573-582.
[29]Langone M,Yan J,Haaijer S C M,et al.Coexistence of nitrifying,anammox and denitrifying bacteria in a sequencing batch reactor[J].Frontiers in Microbiology,2014,5:28.
[30]Du R,Cao S B,Li B K,et al.Simultaneous domestic wastewater and nitrate sewage treatment by Denitrifying Ammonium Oxidation(DEAMOX)in sequencing batch reactor[J].Chemosphere,2017,174:399-407.
[31]Nakasaki K,Tran L T H,Idemoto Y,et al.Comparison of organic matter degradation and microbial community during thermophilic composting of two different types of anaerobic sludge[J].Bioresource Technology,2009,100(2):676-682.
[32]Beer M,Kong Y H,Seviour R J.Are some putative glycogen accumulating organisms(GAO)in anaerobic:aerobic activated sludge systems members of the alpha-Proteobacteria?[J].Microbiology,2004,150(Pt 7):2267-2275.
[2]Li Z M,Wang S Y,Zhang W T,et al.Nitrogen removal from medium-age landfill leachate via post-denitrification driven by PHAs and glycogen in a single sequencing batch reactor[J].Bioresource Technology,2014,169:773-777.
[3]Ciggin A S,Rossetti S,Majone M,et al.Extent of intracellular storage in single and dual substrate systems under pulse feeding[J].Environmental Science and Pollution Research,2013,20(3):1225-1238.
[4]Krasnits E,Beliavski M,Tarre S,et al.Storage-based denitrification with municipal wastewater:influence of the denitrification stage duration[J].Environmental Technology,2014,35(17):2167-2175.
[5]Albuquerque M G E,Concas S,Bengtsson S,et al.Mixed culture polyhydroxyalkanoates production from sugar molasses:The use of a 2-stage CSTR system for culture selection[J].Bioresource Technology,2010,101(18):7112-7122.
[6]Albuquerque M G E,Torres C A V,Reis M A M.Polyhydroxyalkanoate(PHA)production by a mixed microbial culture using sugar molasses:Effect of the influent substrate concentration on culture selection[J].Water Research,2010,44(11):3419-3433.
[7]Pittmann T,Steinmetz H.Polyhydroxyalkanoate production as a side stream process on a municipal waste water treatment plant[J].Bioresource Technology,2014,167:297-302.
[8]Moita R,Freches A,Lemos P C.Crude glycerol as feedstock for polyhydroxyalkanoates production by mixed microbial cultures[J].Water Research,2014,58:9-20.
[9]Chen H B,Yang Q,Li X M,et al.Post-anoxic denitrification via nitrite driven by PHB in feast-famine sequencing batch reactor[J].Chemosphere,2013,92(10):1349-1355.
[10]Miao L,Wang S Y,Li B K,et al.Effect of carbon source type on intracellular stored polymers during endogenous denitritation(ED)treating landfill leachate[J].Water Research,2016,100:405-412.
[11]Albuquerque M G E,Martino V,Pollet E,et al.Mixed culture polyhydroxyalkanoate(PHA)production from volatile fatty acid(VFA)-rich streams:Effect of substrate composition and feeding regime on PHA productivity,composition and properties[J].Journal of Biotechnology,2011,151(1):66-76.
[12]Johnson K,Kleerebezem R,Van Loosdrecht M C M.Influence of the C/N ratio on the performance of polyhydroxybutyrate(PHB)producing sequencing batch reactors at short SRTs[J].Water Research,2010,44(7):2141-2152.
[13]Zhu R L,Wang S Y,Li J,et al.Biological nitrogen removal from landfill leachate using anaerobic-aerobic process:Denitritation via organics in raw leachate and intracellular storage polymers of microorganisms[J].Bioresource Technology,2013,128:401-408.
[14]Eaton A D,Clesceri L S,Greenberg A E,et al.Standard methods for the examination of water and wastewater[M].New York:American Public Health Association,1995.
[15]Cui Y W,Zhang H Y,Lu P F,et al.Effects of carbon sources on the enrichment of halophilic polyhydroxyalkanoate-storing mixed microbial culture in an aerobic dynamic feeding process[J].Scientific Reports,2016,6:30766.
[16]Zeng R J,Van Loosdrecht M C M,Yuan Z G,et al.Metabolic model for glycogen-accumulating organisms in anaerobic/aerobic activated sludge systems[J].Biotechnology and Bioengineering,2003,81(1):92-105.
[17]Cui Y W,Zhang H Y,Ding J R,et al.The effects of salinity on nitrification using halophilic nitrifiers in a Sequencing Batch Reactor treating hypersaline wastewater[J].Scientific Reports,2016,6:24825.
[18]Amato K R,Yeoman C J,Kent A,et al.Habitat degradation impacts black howler monkey(Alouatta pigra)gastrointestinal microbiomes[J].ISME Journal,2013,7(7):1344-1353.
[19]Johnson K,Kleerebezem R,Van Loosdrecht M C M.Influence of ammonium on the accumulation of polyhydroxybutyrate(PHB)in aerobic open mixed cultures[J].Journal of Biotechnology,2010,147(2):73-79.
[20]Beun J J,Paletta F,Van Loosdrecht M C M,et al.Stoichiometry and kinetics of poly-β-hydroxybutyrate metabolism in aerobic,slow growing,activated sludge cultures[J].Biotechnology and Bioengineering,2000,67(4):379-389.
[21]Dionisi D,Majone M,Tandoi V,et al.Sequencing batch reactor:Influence of periodic operation on performance of activated sludges in biological wastewater treatment[J].Industrial&Engineering Chemistry Research,2001,40(23):5110-5119.
[22]Shimada T,Zilles J,Raskin L,et al.Carbohydrate storage in anaerobic sequencing batch reactors[J].Water Research,2007,41(20):4721-4729.
[23]Pittmann T,Steinmetz H.Influence of operating conditions for volatile fatty acids enrichment as a first step for polyhydroxyalkanoate production on a municipal waste water treatment plant[J].Bioresource Technology,2013,148:270-276.
[24]Du R,Cao S B,Niu M,et al.Performance of partialdenitrification process providing nitrite for anammox in sequencing batch reactor(SBR)and upflow sludge blanket(USB)reactor[J].International Biodeterioration&Biodegradation,2017,122:38-46.
[25]Cao S B,Wang S Y,Peng Y Z,et al.Achieving partial denitrification with sludge fermentation liquid as carbon source:The effect of seeding sludge[J].Bioresource Technology,2013,149:570-574.
[26]Ge S J,Peng Y Z,Wang S Y,et al.Nitrite accumulation under constant temperature in anoxic denitrification process:The effects of carbon sources and COD/NO3-N[J].Bioresource Technology,2012,114:137-143.
[27]Ince O,Basak B,Ince B K,et al.Effect of nitrogen deficiency during SBR operation on PHA storage and microbial diversity[J].Environmental Technology,2012,33(16):1827-1837.
[28]Venkateswar Reddy M,Venkata Mohan S.Effect of substrate load and nutrients concentration on the polyhydroxyalkanoates(PHA)production using mixed consortia through wastewater treatment[J].Bioresource Technology,2012,114:573-582.
[29]Langone M,Yan J,Haaijer S C M,et al.Coexistence of nitrifying,anammox and denitrifying bacteria in a sequencing batch reactor[J].Frontiers in Microbiology,2014,5:28.
[30]Du R,Cao S B,Li B K,et al.Simultaneous domestic wastewater and nitrate sewage treatment by Denitrifying Ammonium Oxidation(DEAMOX)in sequencing batch reactor[J].Chemosphere,2017,174:399-407.
[31]Nakasaki K,Tran L T H,Idemoto Y,et al.Comparison of organic matter degradation and microbial community during thermophilic composting of two different types of anaerobic sludge[J].Bioresource Technology,2009,100(2):676-682.
[32]Beer M,Kong Y H,Seviour R J.Are some putative glycogen accumulating organisms(GAO)in anaerobic:aerobic activated sludge systems members of the alpha-Proteobacteria?[J].Microbiology,2004,150(Pt 7):2267-2275.