活性污泥长期好氧饥饿下的微生物种群结构演化
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摘要
为考察好氧饥饿环境对活性污泥硝化及除磷性能的影响,研究活性污泥在长期好氧饥饿条件下的微生物种群结构变化,以具有良好硝化和除磷性能的活性污泥为实验对象,利用Illumina高通量测序平台分别考察活性污泥好氧饥饿处理3,7,14和30 d后的微生物种群结构特性及差异.结果表明:好氧饥饿时间越长,活性污泥硝化及除磷性能所受的影响越大,污泥的种群结构变化越明显.硝化菌和除磷菌等相关功能细菌在短期(7 d)好氧饥饿过程中,可分别利用细菌衰亡裂解释放的氨氮和胞内储能物质进行细胞维持,确保系统硝化及除磷性能的恢复,同时恢复期氨氧化菌快于亚硝酸氧化菌的活性恢复速率促进了系统由全程硝化向短程硝化的转变;而随着好氧饥饿时间的延长,功能细菌的种群丰度均逐渐减少.此外,活性污泥微生物种群结构在30 d好氧饥饿过程中经历了一个动态变化过程,既有优势种群(如Proteobacteria和Bacteroidetes等)的逐步消亡,又有适应好氧饥饿环境的菌种(如Firmicutes)增强成为新的优势菌群.
Aiming at the effect of aerobic starvation environment on the nitrification and phosphorus removal performance of activated sludge and the variations of the microbial community evolution during long-term aerobic starvation periods, an activated sludge with high level removal(>99%) of ammonium and phosphorus was used to study the differences of microbial community structures by Illumina high-throughput sequencing technology during 3, 7, 14, and 30 d aerobic starvation periods. Results showed that the performance of nitrification and phosphorus removal decreased with the increase of the aerobic starvation time, and the change of microbial community structure was more obvious with the proceeding of aerobic starvation. During short-term(7 d) aerobic starvation period, the functional bacteria including nitrifiers and phosphorus removing bacteria utilized the released ammonium from cell decay and intracellular storage polymers for cell maintenance to guarantee the recovery of nitrification and phosphorus removal performance. Besides, higher activity resuscitation rate of ammonium oxidizing bacteria(AOB) than nitrite oxidizing bacteria(NOB) contributed to the shift of the nitrification pathway from nitratation to nitritation. Furthermore, the abundances of functional bacteria gradually decreased with the prolonging of aerobic starvation time. The microbial community structures of the activated sludge underwent a dynamic change during the 30 d aerobic starvation period, in which the majority of the original dominant species within the phylum Proteobacteria and Bacteroidetes declined with a remarkable increase of Firmicutes that could acclimatize themselves to the aerobic starvation conditions.
Aiming at the effect of aerobic starvation environment on the nitrification and phosphorus removal performance of activated sludge and the variations of the microbial community evolution during long-term aerobic starvation periods, an activated sludge with high level removal(>99%) of ammonium and phosphorus was used to study the differences of microbial community structures by Illumina high-throughput sequencing technology during 3, 7, 14, and 30 d aerobic starvation periods. Results showed that the performance of nitrification and phosphorus removal decreased with the increase of the aerobic starvation time, and the change of microbial community structure was more obvious with the proceeding of aerobic starvation. During short-term(7 d) aerobic starvation period, the functional bacteria including nitrifiers and phosphorus removing bacteria utilized the released ammonium from cell decay and intracellular storage polymers for cell maintenance to guarantee the recovery of nitrification and phosphorus removal performance. Besides, higher activity resuscitation rate of ammonium oxidizing bacteria(AOB) than nitrite oxidizing bacteria(NOB) contributed to the shift of the nitrification pathway from nitratation to nitritation. Furthermore, the abundances of functional bacteria gradually decreased with the prolonging of aerobic starvation time. The microbial community structures of the activated sludge underwent a dynamic change during the 30 d aerobic starvation period, in which the majority of the original dominant species within the phylum Proteobacteria and Bacteroidetes declined with a remarkable increase of Firmicutes that could acclimatize themselves to the aerobic starvation conditions.
引文
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[16]LU Huabing,KELLER J,YUAN Zhiguo.Endogenous metabolism of Candidatus Accumulibacter phosphatis under various starvation conditions[J].Water Research,2007,41:4646.DOI:10.1016/j.watres.2007.06.046
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[18]LU Lu,XING Defeng,REN Nanqi.Pyrosequencing reveals highly diverse microbial communities in microbial electrolysis cells involved in enhanced H2 production from waste activated sludge[J].Water Research,2012,46(7):2425.DOI:10.1016/j.watres.2012.02.005
[19]WETT B,RAUCH W.The role of inorganic carbon limitation in biological nitrogen removal of extremely ammonia concentrated wastewater[J].Water Research,2003,37(5):1100.DOI:10.1016/S0043-1354(2)00 440-2
[2]ELAWWAD A,SANDNER H,KAPPELMEYER U,et al.Long-term starvation and subsequent recovery of nitrifiers in aerated submerged fixed-bed biofilm reactors[J].Environmental Technology,2013,34(8):945.DOI:10.1080/09593330.2012.722758
[3]胡怡杉,孙宝盛,王盛勇.MBR和CAS工艺污泥在贫营养培养条件下的微生物群落结构研究[J].环境科学学报,2011,31(9):1900HU Yishan,SUN Baosheng,WANG Shengyong.Microbial community structure of sludge in MBR and CAS under oligotrophic environment[J].Acta Scientiae Circumstantiae,2011,31(9):1900
[4]WANG Bo,PENG Yongzhen,GUO Yuanyuan,et al.Illumina MiSeq sequencing reveals the key microorganisms involved in partial nitritation followed by simultaneous sludge fermentation,denitrification and anammox process[J].Bioresource Technology,2016,207:118.DOI:10.1016/j.biortech.2016.01.072
[5]YE Lin,SHAO Mingfei,ZHANG Tong,et al.Analysis of the bacterial community in a laboratory-scale nitrification reactor and a wastewater treatment plant by 454-pyrosequencing[J].Water Research,2011,45(15):4390.DOI:10.1016/j.watres.2011.05.028
[6]LIU Wenlong,YANG Qing,MA Bin,et al.Rapid achievement of nitritation using aerobic starvation[J].Environmental Science & Technology,2017,51:4001.DOI:10.1021/acs.est.6b04598
[7]Federation Water Environmental.Standard methods for the examination of water and wastewater[M].Washington D C:American Public Health Association (APHA),2005
[8]BOLLMANN A,SCHMIDT I,SAUNDSES A M,et al.Influence of starvation on potential ammonia-oxidizing activity and amoA mRNA levels of Nitrosospira briensis[J].Applied and Environmental Microbiology,2005,71(3):1276.DOI:10.1128/AEM.71.3.1276-1282.2005
[9]SMITH N R,YU Zhongtang,MOHN W W.Stability of the bacterial community in a pulp mill effluent treatment system during normal operation and a system shutdown[J].Water Research,2003,37(20):4873.DOI:10.1016/j.watres.2003.08.019
[10]LIANG Bin,CHENG Haoyi,VAN NOSTRAND J D,et al.Microbial community structure and function of nitrobenzene reduction biocathode in response to carbon source switchover[J].Water Research,2014,54:137.DOI:10.1016/j.watres.2014.01.052
[11]ZHANG Tong,SHAO Mingfei,YE Lin.454 pyrosequencing reveals bacterial diversity of activated sludge from 14 sewage treatment plants[J].The ISME Journal,2012,6:1137.DOI:10.1038/ismej.2011.188
[12]WEISSBRODT D G,MAILLARD J,BROVELLI A,et al.Multilevel correlations in the biological phosphorus removal process:From bacterial enrichment to conductivity-based metabolic batch tests and polyphosphatase assays[J].Biotechnology and Bioengineering,2014,111(12):2421.DOI:10.1002/bit.25320
[13]PITERINA A V,BARTLETT J,PEMBROKE J T.Phylogenetic analysis of the bacterial community in a full scale autothermal thermophilic aerobic digester (ATAD) treating mixed domestic wastewater sludge for land spread[J].Water Research,2012,46(8):2488.DOI:10.1016/j.watres.2012.01.045
[14]LI Dong,LV Yufeng,ZENG Huiping,et al.Long term operation of continuous-flow system with enhanced biological phosphorus removal granules at different COD loading[J].Bioresource Technology,2016,216:761.DOI:10.1016/j.biortech.2016.06.022
[15]ZHANG Miao,YANG Qing,ZHANG Jianhua,et al.Enhancement of denitrifying phosphorus removal and microbial community of long-term operation in an anaerobic anoxic oxic-biological contact oxidation system[J].Journal of Bioscience and Bioengineering,2016,122(4):456.DOI:10.1016/j.jbiosc.2016.03.019
[16]LU Huabing,KELLER J,YUAN Zhiguo.Endogenous metabolism of Candidatus Accumulibacter phosphatis under various starvation conditions[J].Water Research,2007,41:4646.DOI:10.1016/j.watres.2007.06.046
[17]MCLLROY S J,STARNAWSKA A,STARNAWSKI P,et al.Identification of active denitrifiers in full-scale nutrient removal wastewater treatment systems[J].Environment Microbiology,2014,18(1):50.DOI:10.1111/1462-2920.12614
[18]LU Lu,XING Defeng,REN Nanqi.Pyrosequencing reveals highly diverse microbial communities in microbial electrolysis cells involved in enhanced H2 production from waste activated sludge[J].Water Research,2012,46(7):2425.DOI:10.1016/j.watres.2012.02.005
[19]WETT B,RAUCH W.The role of inorganic carbon limitation in biological nitrogen removal of extremely ammonia concentrated wastewater[J].Water Research,2003,37(5):1100.DOI:10.1016/S0043-1354(2)00 440-2