新型A/O/A/O-SBR中DPB的富集及其典型菌株特性研究
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摘要
为能更好地达到同步除磷脱氮的目的,对反硝化聚磷菌(Den-itrifying Phosphorus Removal Bacteria,DPB)进行了富集培养,并对其中的典型菌株进行了特性研究。以校园生活区污水为研究对象,在温度为25℃,pH值为7.5,乙酸钠为碳源,进水COD为316.5 mg/L的条件下,采用三阶段的污泥驯化,对反硝化聚磷菌种进行了分离纯化,并对富集的典型菌株进行了生理生化试验、吸磷试验、硝酸盐还原产气试验。结果表明,富集培养后的DPB在A/O/A/O-SBR系统(厌氧2 h,好氧1.5 h,缺氧1.5 h,后置曝气0.5 h)中成为优势菌群,系统中COD、NH4+-N、TN、TP的出水质量浓度分别为28.35 mg/L、0.87 mg/L、4.05mg/L和0.37 mg/L。分离鉴定出具有反硝化除磷能力的Z7、Z9两株典型菌株,其吸磷率均在50%左右。经细菌形态观察、生理特性分析及16S rDNA序列测定,鉴定Z7、Z9为缺氧反硝化聚磷菌,与假单胞菌(Pseudomonas)最相似,其同源性均达99.9%。
The present paper pursues a study of the enrichment way and characteristic features of typical DPB strains in a new species of A/O/A/O-SBR.The research urgency has come down to us is that in recent years,the land freshwater quality has been worsening than before due to the serious water eutrophication and greater nitrogen and phosphorus content in the sewage,which makes it an essential problem for water quality control.To achieve the synchronous phosphorus removal and denitrification,the present paper is dedicated to the study of the accumulation culture of the denitrifying phosphorus removal bacteria(DPB)and the characteristic features of the typical strains in a new species of anaerobic/aerobic/anoxic/aerobic SBR.In our study,we have done purifying treatment of the sewage of campus living area and conducted experiment with the carbon source of sodium acetate with the mass density of the water inlet of COD of 316.5 mg/L in an Sequencing Batch Reactor at the temperature of 25 ℃ and pH value of 7.5.Our experiment was started with the three-phase-training method,trying to remove bacteria from the mixed liquor samples in the system by isolating the denitrifying phosphorus,which was followed by a series of physiological and biochemical experiments,the denitrification P-uptake experiments and those of denitrifying nitrates to gas,and so on and so forth.The results of our experimental research prove that the phosphorus removal bacteria were enriched with a new species of A/O/A/O-SBR with anaerobic 2 h,aerobic 1.5 h,anoxia 1.5 h,aerobic 0.5 h.And,in turn,the A/O/A/O-SBR system of the COD,NH+4-N,TN,the percentage of TP removal were found equal to 91%,97%,88% and 93%,respectively,with the effluent mass density being 28.35 mg/L,0.87 mg/L,4.05 mg/L and 0.37 mg/L,respectively.Repeated processing,such as separation and purification would let us obtain Z7 to Z9 strains polyphosphate accumulation bacteria in an SBR system,which belong to the gram-negative bacteria,both of which contain metachromatic granules.And both of the strains can release and absorb phosphorus,whose absorbing rate is 50%.In addition,the two strains can also help to treat nitrate-reducing aerogenesis.They are also taken as two kinds of denitrifying phosphorus removal bacteria.Careful morphological observation,physiological characteristics analysis and 16S rDNA sequential examination prove that they are most similar to Pseudomonas,with 99.9% homology of sequence.
The present paper pursues a study of the enrichment way and characteristic features of typical DPB strains in a new species of A/O/A/O-SBR.The research urgency has come down to us is that in recent years,the land freshwater quality has been worsening than before due to the serious water eutrophication and greater nitrogen and phosphorus content in the sewage,which makes it an essential problem for water quality control.To achieve the synchronous phosphorus removal and denitrification,the present paper is dedicated to the study of the accumulation culture of the denitrifying phosphorus removal bacteria(DPB)and the characteristic features of the typical strains in a new species of anaerobic/aerobic/anoxic/aerobic SBR.In our study,we have done purifying treatment of the sewage of campus living area and conducted experiment with the carbon source of sodium acetate with the mass density of the water inlet of COD of 316.5 mg/L in an Sequencing Batch Reactor at the temperature of 25 ℃ and pH value of 7.5.Our experiment was started with the three-phase-training method,trying to remove bacteria from the mixed liquor samples in the system by isolating the denitrifying phosphorus,which was followed by a series of physiological and biochemical experiments,the denitrification P-uptake experiments and those of denitrifying nitrates to gas,and so on and so forth.The results of our experimental research prove that the phosphorus removal bacteria were enriched with a new species of A/O/A/O-SBR with anaerobic 2 h,aerobic 1.5 h,anoxia 1.5 h,aerobic 0.5 h.And,in turn,the A/O/A/O-SBR system of the COD,NH+4-N,TN,the percentage of TP removal were found equal to 91%,97%,88% and 93%,respectively,with the effluent mass density being 28.35 mg/L,0.87 mg/L,4.05 mg/L and 0.37 mg/L,respectively.Repeated processing,such as separation and purification would let us obtain Z7 to Z9 strains polyphosphate accumulation bacteria in an SBR system,which belong to the gram-negative bacteria,both of which contain metachromatic granules.And both of the strains can release and absorb phosphorus,whose absorbing rate is 50%.In addition,the two strains can also help to treat nitrate-reducing aerogenesis.They are also taken as two kinds of denitrifying phosphorus removal bacteria.Careful morphological observation,physiological characteristics analysis and 16S rDNA sequential examination prove that they are most similar to Pseudomonas,with 99.9% homology of sequence.
引文
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[9]WANG Chunli(王春丽),MA Fang(马放),LIU Hui(刘慧),et al.Screening and isolating method for denitrifying polyphosphate-accumulat-ing organisms[J].Journal of Jilin Architectural and Civil EngineeringInstitute(吉林建筑工程学院学报),2006,23(3):5-8.
[10]DONG Xiuzhu(东秀珠),CAI Miaoying(蔡妙英).Identification ofcommon bacterial systems manual(常见细菌系统鉴定手册)[M].Beijing:Science Press,2001:349-364.
[11]LI Yafeng(李亚峰),HAO Ying(郝滢),ZHANG Xiaoning(张晓宁),et al.Study on Separation and Identification of denitrifying phos-phorus removing bacteria in SBBR[J].Journal of Shenyang JianzhuUniversity:Natural Science(沈阳建筑大学学报:自然科学版),2011,27(5):952-956.
[2]MERZOUKIM,BERNET N,DELGENES J P,et al.Biological denitrify-ing phosphorus removal in SBR:effect of added nitrate concentration andsludge retention time[J].Water Science andTechnology,2001,43(3):191-194.
[3]HE S,GU A,McMAHON Z K D.Fine scale differences between accu-mulibacter-like bacteria in enhanced biological phosphorus removal act-ivated sludge[J].Water Science and Technology,2006,54(1):111-117.
[4]LI Yongzhi(李勇智),PENGYongzhen(彭永臻),WANGShuying(王淑滢).Denitrifying phosphorus removal by anaerobic/anoxic sequenc-ing batch reactor[J].Techniques and Equipmentfor Environmental Pol-lution Control(环境污染治理技术与设备),2003,16(1):9-12.
[5]SHI Yuming(石玉明),MA Fang(马放),WANG Qiang(王强),et al.Isolation of PAOs and efficiency of phosphorous accumulation in SBRsystem[J].Journal of Harbin Institute of Technology(哈尔滨工业大学学报),2008,40(12):1927-1931.
[6]LIU Hui(刘晖),SUN Yanfu(孙彦富),CUI Yingde(崔英德),etal.Identification of a shortcut denitrifying phosphorus removing bacterialstrain and its biological utilization[J].Journal of Chemical Industry andEngineering(China)(化工学报),2009,60(7):1758-1766.
[7]JIAO Zhongzhi(焦中志),LI Xiangkun(李相昆),ZHANG Licheng(张立成),et al.Isolation and characteristic of denitrifying phosphorusaccumulating organism[J].Journal of Shenyang Jianzhu University:Natural Science(沈阳建筑大学学报:自然科学版),2009,25(3):535-540.
[8]LIU Hui(刘晖),SUN Yanfu(孙彦富),ZHOU Kangqun(周康群),et al.Screening and growth condition optimization of anoxic denitrifyingphosphorus removal bacteria Xun X[J].Research of Environmental Sci-ences(环境科学研究),2010,23(2):203-208.
[9]WANG Chunli(王春丽),MA Fang(马放),LIU Hui(刘慧),et al.Screening and isolating method for denitrifying polyphosphate-accumulat-ing organisms[J].Journal of Jilin Architectural and Civil EngineeringInstitute(吉林建筑工程学院学报),2006,23(3):5-8.
[10]DONG Xiuzhu(东秀珠),CAI Miaoying(蔡妙英).Identification ofcommon bacterial systems manual(常见细菌系统鉴定手册)[M].Beijing:Science Press,2001:349-364.
[11]LI Yafeng(李亚峰),HAO Ying(郝滢),ZHANG Xiaoning(张晓宁),et al.Study on Separation and Identification of denitrifying phos-phorus removing bacteria in SBBR[J].Journal of Shenyang JianzhuUniversity:Natural Science(沈阳建筑大学学报:自然科学版),2011,27(5):952-956.