内源碳PHA的贮存对混合菌群耐低温特性的影响
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摘要
本文研究碳源调控磷回收强化生物脱氮除磷(biological bio-nutrient removal-carbon regulation and phosphorus recovery,BBNR-CPR)反应器耐低温特性.不断降低BBNR-CPR反应器运行温度,发现BBNR-CPR反应器能够长期在低温(≤15℃)、低C/N比(<4. 16)条件下稳定运行,维持总磷的平均去除率为91. 20%,氨氮的平均去除率为81. 10%,总氮的平均去除率为58. 62%.随着运行时间的增加与温度的下降,BBNR-CPR反应器生物内膜具有脱氮除磷以及PHA贮存功能的种属Candidatus_Competibacter、Candidatus_Accumulibacter、Run-SP154、Thauera、Candidatus_Nitrotoga的相对丰度不断提高,成为耐受低温条件的优势种属.在相同碳源浓度、合成PHA的时间一致的条件下,生物膜内PHA的合成量受温度影响; 25、15和8℃合成的PHA量分别占生物膜干重的16. 24%、11. 49%和9. 01%.预贮存PHA的生物膜具有耐受低温的能力;在高PHA水平,8℃和15℃的除磷效率分别为97. 46%和100%,脱氮效率分别为55. 15%和82. 55%;而在低PHA水平,8℃和15℃的除磷效率分别为11. 39%和35. 02%,脱氮效率分别为0%和12. 10%.
This study investigated the removal of nitrogen and phosphorous by a biological bio-nutrient removal-carbon regulation and phosphorus recovery( BBNR-CPR) reactor at low temperature. The operating temperature of the BBNR-CPR reactor was continuously reduced,and it was found that the BBNR-CPR reactor could operate steadily at low temperature( < 15℃) and low C/N ratio( < 4. 16). The average removal rates of total phosphorus,ammonia,and total nitrogen were 91. 20%,81. 10%,and 58. 62%,respectively. With increasing running time and decreasing temperature, the relative abundance of Candidatus _ Competibacter,Candidatus_Accumulibacter,Run-SP154,Thauera,and Candidatus_Nitrotoga increased. These bacteria had the functions of nitrogen and phosphorus removal and the storage of polyhydroxyalkanoate( PHA) in the biofilm of the BBNR-CPR reactor and became the dominant species to tolerate low temperature. It was found that low temperatures reduced the amount of PHA synthesized for a given equal carbon source concentration and reaction time. The amounts of PHA synthesized at 25℃,15℃,and 8℃ accounted for16. 24%,11. 49%,and 9. 01% of the dry weight of the biofilm,respectively. The pre-stored PHA biofilm has the capacity to resist low temperature. At high PHA levels,the phosphorus removal efficiencies at 8℃ and 15℃ were 97. 46% and 100%,respectively,and the denitrification efficiencies were 55. 15% and 82. 55%. At low PHA levels,the phosphorus removal efficiencies at 8℃ and15℃ were 11. 39% and 35. 02%,respectively,and the denitrification efficiencies were 0% and 12. 10%,respectively.
This study investigated the removal of nitrogen and phosphorous by a biological bio-nutrient removal-carbon regulation and phosphorus recovery( BBNR-CPR) reactor at low temperature. The operating temperature of the BBNR-CPR reactor was continuously reduced,and it was found that the BBNR-CPR reactor could operate steadily at low temperature( < 15℃) and low C/N ratio( < 4. 16). The average removal rates of total phosphorus,ammonia,and total nitrogen were 91. 20%,81. 10%,and 58. 62%,respectively. With increasing running time and decreasing temperature, the relative abundance of Candidatus _ Competibacter,Candidatus_Accumulibacter,Run-SP154,Thauera,and Candidatus_Nitrotoga increased. These bacteria had the functions of nitrogen and phosphorus removal and the storage of polyhydroxyalkanoate( PHA) in the biofilm of the BBNR-CPR reactor and became the dominant species to tolerate low temperature. It was found that low temperatures reduced the amount of PHA synthesized for a given equal carbon source concentration and reaction time. The amounts of PHA synthesized at 25℃,15℃,and 8℃ accounted for16. 24%,11. 49%,and 9. 01% of the dry weight of the biofilm,respectively. The pre-stored PHA biofilm has the capacity to resist low temperature. At high PHA levels,the phosphorus removal efficiencies at 8℃ and 15℃ were 97. 46% and 100%,respectively,and the denitrification efficiencies were 55. 15% and 82. 55%. At low PHA levels,the phosphorus removal efficiencies at 8℃ and15℃ were 11. 39% and 35. 02%,respectively,and the denitrification efficiencies were 0% and 12. 10%,respectively.
引文
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[12] Pradhan S,Borah AJ,Poddar M K,et al. Microbial production,ultrasound-assisted extraction and characterization of biopolymer polyhydroxybutyrate(PHB)from terrestrial(P. hysterophorus)and aquatic(E. crassipes)invasive weeds[J]. Bioresource Technology,2017,242:304-310.
[13] Inoue D,Suzuki Y,Sawada K,et al. Polyhydroxyalkanoate accumulation ability and associated microbial community in activated sludge-derived acetate-fed microbial cultures enriched under different temperature and pH conditions[J]. Journal of Bioscience and Bioengineering,2017,125(3):329-345.
[14] Liu S L,Li J Z. Accumulation and isolation of simultaneous denitrifying polyphosphate-accumulating organisms in an improved sequencing batch reactor system at low temperature[J].International Biodeterioration and Biodegradation,2015,100:140-148.
[15] Laureni M, Falas P, Robin O, et al. Mainstream partial nitritation and anammox:long-term process stability and effluent quality at low temperatures[J]. Water Research,2016,101:628-639.
[16]金羽.温度对A2/O系统的影响特征及脱氮除磷强化技术研究[D].哈尔滨:哈尔滨工业大学,2013. 15-20.Jin Y. Effection of temperature on the performance of an A2/O process and enhancement of nitrogen and phosphorus removal[D]. Harbin:Harbin Institute of Technology,2013. 15-20.
[17] Helmer C,Kunst S. Low temperature effects on phosphorus release and uptake by microorganisms in ebpr plants[J]. Water Science and Technology,1998,37(4-5):531-539.
[18] Jiang Y,Marang L,Kleerebezem R,et al. Effect of temperature and cycle length on microbial competition in PHB-producing sequencing batch reactor[J]. The ISME Journal,2011,5(5):896-907.
[19] Inoue D,Suzuki Y,Uchida T,et al. Polyhydroxyalkanoate production potential of heterotrophic bacteria in activated sludge[J]. Journal of Bioscience and Bioengineering,2015,121(1):47-51.
[20] Johnson K,Van Geest J,Kleerebezem R,et al. Short-and longterm temperature effects on aerobic polyhydroxybutyrate producing mixed cultures[J]. Water Research,2010,44(6):1689-1700.
[21] Tian Q,Ong S K,Xie X H,et al. Enhanced phosphorus recovery and biofilm microbial community changes in an alternating anaerobic/aerobic biofilter[J]. Chemosphere,2016,144:1797-1806.
[22]魏复盛.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. 266-268.
[23] Tan G Y A, Chen C L, Ge L Y, et al. Enhanced gas chromatography-mass spectrometry method for bacterial polyhydroxyalkanoates analysis[J]. Journal of Bioscience and Bioengineering,2014,117(3):379-382.
[24] Seviour R J,Mino T,Onuki M. The microbiology of biological phosphorus removal in activated sludge systems[J]. FEMS Microbiology Reviews,2010,27(1):99-127.
[25] Mcilroy S J,Albertsen M,Andresen E K,et al.‘Candidatus Competibacter’-lineage genomes retrieved from metagenomes reveal functional metabolic diversity[J]. The ISME Journal,2014,8(3):613-624.
[26] Marang L,Van Loosdrecht M C M,Kleerebezem R. Enrichment of PHA-producing bacteria under continuous substrate supply[J]. New Biotechnology,2017,41:55-61.
[27] Alawi M,Lipski A,Sanders T,et al. Cultivation of a novel coldadapted nitrite oxidizing betaproteobacterium from the Siberian Arctic[J]. The ISME Journal,2007,1(3):256-264.
[28] Lücker S,Schwarz J,Gruber-Dorninger C,et al. Nitrotoga-like bacteria are previously unrecognized key nitrite oxidizers in fullscale wastewater treatment plants[J]. The ISME Journal,2015,9(3):708-720.
[29] Zhou H X,Li X,Xu G R,et al. Overview of strategies for enhanced treatment of municipal/domestic wastewater at low temperature[J]. Science of the Total Environment,2018,643:225-237.
[30] 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.
[31] Zhao X Y,Yang J X,Bai S W,et al. Microbial population dynamics in response to bioaugmentation in a constructed wetland system under 10℃[J]. Bioresource Technology,2016,205:166-173.
[32] Kouba V,VejmelkováD,Proksova E,et al. High-rate partial nitritation of municipal wastewater after psychrophilic anaerobic pretreatment[J]. Environmental Science&Technology,2017,51(19):11029-11038.
[2] Lei M,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.
[3] Huang L,Chen Z Q,Wen Q X,et al. Insights into feast-famine polyhydroxyalkanoate(PHA)-producer selection:Microbial community succession,relationships with system function and underlying driving forces[J]. Water Research,2018,131:167-176.
[4]姚樱,陈银广,马民,等.不同丙酸/乙酸对聚糖菌长/短期代谢的影响[J].环境科学,2007,28(9):1970-1974.Tao Y,Chen Y G,Ma M,et al. Long-term and short-term effects of propionic/acetic acid ratios on metabolism of glycogenaccumulating organisms[J]. Environmental Science,2007,28(9):1970-1974.
[5] Tian Q,Zhuang L J,Ong S K,et al. Phosphorus(P)recovery coupled with increasing influent ammonium facilitated intracellular carbon source storage and simultaneous aerobic phosphorus&nitrogen removal[J]. Water Research,2017,119:267-275.
[6] Wang X X,Wang S Y,Xue T L,et al. Treating low carbon/nitrogen(C/N)wastewater in simultaneous nitrificationendogenous denitrification and phosphorous removal(SNDPR)systems by strengthening anaerobic intracellular carbon storage[J]. Water Research,2015,77:191-200.
[7] Zeng R J, Lemaire R, Yuan Z G, et al. Simultaneous nitrification,denitrification,and phosphorus removal in a labscale sequencing batch reactor[J]. Biotechnology and Bioengineering,2003,84(2):170-178.
[8] Saad S A,Welles L,Abbas B,et al. Denitrification of nitrate and nitrite by‘Candidatus Accumulibacter phosphatis’clade IC[J]. Water Research,2016,105:97-109.
[9] Wang X X,Wang S Y,Zhao J,et al. Combining simultaneous nitrification-endogenous denitrification and phosphorus removal with post-denitrification for low carbon/nitrogen wastewater treatment[J]. Bioresource Technology,2016,220:17-25.
[10] Miao L,Wang S Y,Li B K,et al. Advanced nitrogen removal via nitrite using stored polymers in a modified sequencing batch reactor treating landfill leachate[J]. Bioresource Technology,2015,192:354-360.
[11] Ji J T,Peng Y Z,Bo W,et al. Achievement of high nitrite accumulation via endogenous partial denitrification(EPD)[J].Bioresource Technology,2016,224:140-146.
[12] Pradhan S,Borah AJ,Poddar M K,et al. Microbial production,ultrasound-assisted extraction and characterization of biopolymer polyhydroxybutyrate(PHB)from terrestrial(P. hysterophorus)and aquatic(E. crassipes)invasive weeds[J]. Bioresource Technology,2017,242:304-310.
[13] Inoue D,Suzuki Y,Sawada K,et al. Polyhydroxyalkanoate accumulation ability and associated microbial community in activated sludge-derived acetate-fed microbial cultures enriched under different temperature and pH conditions[J]. Journal of Bioscience and Bioengineering,2017,125(3):329-345.
[14] Liu S L,Li J Z. Accumulation and isolation of simultaneous denitrifying polyphosphate-accumulating organisms in an improved sequencing batch reactor system at low temperature[J].International Biodeterioration and Biodegradation,2015,100:140-148.
[15] Laureni M, Falas P, Robin O, et al. Mainstream partial nitritation and anammox:long-term process stability and effluent quality at low temperatures[J]. Water Research,2016,101:628-639.
[16]金羽.温度对A2/O系统的影响特征及脱氮除磷强化技术研究[D].哈尔滨:哈尔滨工业大学,2013. 15-20.Jin Y. Effection of temperature on the performance of an A2/O process and enhancement of nitrogen and phosphorus removal[D]. Harbin:Harbin Institute of Technology,2013. 15-20.
[17] Helmer C,Kunst S. Low temperature effects on phosphorus release and uptake by microorganisms in ebpr plants[J]. Water Science and Technology,1998,37(4-5):531-539.
[18] Jiang Y,Marang L,Kleerebezem R,et al. Effect of temperature and cycle length on microbial competition in PHB-producing sequencing batch reactor[J]. The ISME Journal,2011,5(5):896-907.
[19] Inoue D,Suzuki Y,Uchida T,et al. Polyhydroxyalkanoate production potential of heterotrophic bacteria in activated sludge[J]. Journal of Bioscience and Bioengineering,2015,121(1):47-51.
[20] Johnson K,Van Geest J,Kleerebezem R,et al. Short-and longterm temperature effects on aerobic polyhydroxybutyrate producing mixed cultures[J]. Water Research,2010,44(6):1689-1700.
[21] Tian Q,Ong S K,Xie X H,et al. Enhanced phosphorus recovery and biofilm microbial community changes in an alternating anaerobic/aerobic biofilter[J]. Chemosphere,2016,144:1797-1806.
[22]魏复盛.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. 266-268.
[23] Tan G Y A, Chen C L, Ge L Y, et al. Enhanced gas chromatography-mass spectrometry method for bacterial polyhydroxyalkanoates analysis[J]. Journal of Bioscience and Bioengineering,2014,117(3):379-382.
[24] Seviour R J,Mino T,Onuki M. The microbiology of biological phosphorus removal in activated sludge systems[J]. FEMS Microbiology Reviews,2010,27(1):99-127.
[25] Mcilroy S J,Albertsen M,Andresen E K,et al.‘Candidatus Competibacter’-lineage genomes retrieved from metagenomes reveal functional metabolic diversity[J]. The ISME Journal,2014,8(3):613-624.
[26] Marang L,Van Loosdrecht M C M,Kleerebezem R. Enrichment of PHA-producing bacteria under continuous substrate supply[J]. New Biotechnology,2017,41:55-61.
[27] Alawi M,Lipski A,Sanders T,et al. Cultivation of a novel coldadapted nitrite oxidizing betaproteobacterium from the Siberian Arctic[J]. The ISME Journal,2007,1(3):256-264.
[28] Lücker S,Schwarz J,Gruber-Dorninger C,et al. Nitrotoga-like bacteria are previously unrecognized key nitrite oxidizers in fullscale wastewater treatment plants[J]. The ISME Journal,2015,9(3):708-720.
[29] Zhou H X,Li X,Xu G R,et al. Overview of strategies for enhanced treatment of municipal/domestic wastewater at low temperature[J]. Science of the Total Environment,2018,643:225-237.
[30] 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.
[31] Zhao X Y,Yang J X,Bai S W,et al. Microbial population dynamics in response to bioaugmentation in a constructed wetland system under 10℃[J]. Bioresource Technology,2016,205:166-173.
[32] Kouba V,VejmelkováD,Proksova E,et al. High-rate partial nitritation of municipal wastewater after psychrophilic anaerobic pretreatment[J]. Environmental Science&Technology,2017,51(19):11029-11038.