反硝化深床滤柱深度脱氮效果及反硝化功能基因分析
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摘要
采用石英砂、活性炭双层滤料反硝化深床滤柱处理城镇污水处理厂二沉池出水中硝酸盐氮(NO3–-N),研究了深床滤柱反硝化脱氮性能以及主要反硝化功能基因对进水碳氮比(化学需氧量/总氮,即COD/TN,简称C/N)的响应。结果表明,NO3–-N平均转化率随着C/N升高,由46.5%升高至90.0%,化学需氧量(COD)平均去除率由97.2%降至76.5%。低碳氮比(C/N<6)条件下,出水亚硝酸盐氮(NO2–-N)出现明显的积累,在C/N=4.2时,积累率达41.5%,在高碳氮比(C/N≥6)条件下,NO2–-N积累量逐渐减少,直至出水无NO2–-N。研究表明,反硝化深床滤柱对污染物的转化主要发生在前35cm滤料深度,COD去除率和NO3–-N转化率分别为94.0%、81.2%。采用荧光实时定量PCR技术在C/N分别为4.2、6和7条件下,对深床滤柱中反硝化功能基因napA、narG、nirK、nirS和nosZ数量进行分析,结果表明,随着C/N升高,各反硝化功能基因拷贝数也随之升高,说明增加碳源投加量可以为反硝化细菌提供更好的生长环境,有利于其生长繁殖,促进反硝化过程的进行;当narG基因拷贝数大于(nirS+nirK)基因拷贝数时,NO2–-N会产生积累。
The denitrification performance and the change of denitrifying genes under different chemical oxygen demand(COD)/total nitrogen(TN)(C/N) conditions in a deep bed denitrification filter which used dual medium of quartz sand and activated carbon were investigated, when the filter treated the nitrate(NO3–-N) of effluent from the secondary sedimentation tank in urban wastewater treatment plant. The result showed that the average of NO3–-N conversion rate and COD removal efficiency increased from 46.5% to 90.0% and from 97.2% to 76.5%, respectively. Nitrite(NO2–-N) accumulated under low C/N(C/N<6). When C/N was 4.2, NO2–-N accumulation ratio reached 41.5%. With the increase of C/N(C/N≥6), the accumulation of NO2–-N gradually decreased until no NO2–-N was detected in the effluent of the filter. The pollutants removal in the deep-bed denitrification filter mainly occurred in the first 35 cm, which allowed for COD removal efficiency and NO3–-N conversion rate of 94.0% and 81.2%, respectively. When C/N was 4.2, 6 and 7, the abundance of napA, narG, nirK, nirS and nosZ genes in the deep-bed denitrification filter was investigated by real-time quantitativePCR(qPCR). The number of denitrifying genes copies increased with the increase of C/N. It was found that the increase of carbon source could provide better environment for denitrifying bacteria and be beneficial to the growth and reproduction of denitrifying bacteria and then promote the denitrification process. Moreover, it was concluded that when the number of narG copies was higher than the sum of nirS and nirK copies, there was nitrite accumulation.
The denitrification performance and the change of denitrifying genes under different chemical oxygen demand(COD)/total nitrogen(TN)(C/N) conditions in a deep bed denitrification filter which used dual medium of quartz sand and activated carbon were investigated, when the filter treated the nitrate(NO3–-N) of effluent from the secondary sedimentation tank in urban wastewater treatment plant. The result showed that the average of NO3–-N conversion rate and COD removal efficiency increased from 46.5% to 90.0% and from 97.2% to 76.5%, respectively. Nitrite(NO2–-N) accumulated under low C/N(C/N<6). When C/N was 4.2, NO2–-N accumulation ratio reached 41.5%. With the increase of C/N(C/N≥6), the accumulation of NO2–-N gradually decreased until no NO2–-N was detected in the effluent of the filter. The pollutants removal in the deep-bed denitrification filter mainly occurred in the first 35 cm, which allowed for COD removal efficiency and NO3–-N conversion rate of 94.0% and 81.2%, respectively. When C/N was 4.2, 6 and 7, the abundance of napA, narG, nirK, nirS and nosZ genes in the deep-bed denitrification filter was investigated by real-time quantitativePCR(qPCR). The number of denitrifying genes copies increased with the increase of C/N. It was found that the increase of carbon source could provide better environment for denitrifying bacteria and be beneficial to the growth and reproduction of denitrifying bacteria and then promote the denitrification process. Moreover, it was concluded that when the number of narG copies was higher than the sum of nirS and nirK copies, there was nitrite accumulation.
引文
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[15]BRU D,SARR A,PHILIPPOT L.Relative abundances of proteobacterial membrane-bound and periplasmic nitrate reductases in selected environments[J].Applied&Environmental Microbiology,2007,73(18):5971-5974.
[16]MICHOTEY V,MéJEAN V,BONIN P.Comparison of methods for quantification of cytochrome CD1-denitrifying bacteria in environmental marine samples[J].Applied&Environmental Microbiology,2000,66(4):1564-1571.
[17]HALLIN S,LINDGREN P E.PCR detection of genes encoding nitrite reductase in denitrifying bacteria[J].Applied&Environmental Microbiology,1999,65(4):1652-1657.
[18]HENRY S,BRU D,STRES B,et al.Quantitative detection of the nosZ gene,encoding nitrous oxide reductase,and comparison of the abundances of 16S rRNA,narG,nirK,and nosZ genes in soils[J].Applied&Environmental Microbiology,2006,72(8):5181-5189.
[19]吴兴海,李咏梅.碳氮比对不同滤料反硝化滤池脱氮效果的影响[J].环境工程学报,2017(1):55-62.WU Xinghai,LI Yongmei.Effect of C/N ratio in denitrification of denitrification filters with different filter materials[J].Chinese Journal of Environmental Engineering,2017(1):55-62.
[20]HOU J,XIA L,MA T,et al.Achieving short-cut nitrification and denitrification in modified intermittently aerated constructed wetland[J].Bioresource Technology,2017,232:10.
[21]葛勇涛,焦阳,李军,等.乙酸钠碳源强化生物滤池对二沉池出水的脱氮效果[J].中国给水排水,2011,27(7):98-100.GE Yongtao,JIAO Yang,LI Jun,et al.Sodium acetate as carbon source for enhancing nitrogen removal from secondary effluent with biological filter[J].China Water and Wastewater,2011,27(7):98-100.
[22]HER J J,HUANG J S.Influences of carbon source and C/N ratio on nitrate/nitrite denitrification and carbon breakthrough[J].Bioresource Technology,1995,54(1):45-51.
[23]邹联沛,王宝贞,迟军,等.膜生物反应器处理污水性能的研究[J].哈尔滨建筑大学学报,2001,34(2):57-60.ZOU Lianpei,WANG Baozhen,CHI Jun,et al.Wastewater treating capability of membrane bioreactor[J].Journal of Harbin University of Civil Engineering And Architecture,2001,34(2):57-60.
[24]LEVY-BOOTH D J,PRESCOTT C E,GRAYSTON S J.Microbial functional genes involved in nitrogen fixation,nitrification and denitrification in forest ecosystems[J].Soil Biology&Biochemistry,2014,75:11-25.
[25]张盛博,何小娟,吴海露,等.分子生态学技术用于反硝化微生物群落研究的进展[J].环境科学与技术,2017(6):120-126.ZHANG Shengbo,HE Xiaojuan,WU Hailu,et al.Advances in molecular ecology technologies for the study of denitrifier communities[J].Environmental Science and Technology,2017(6):120-126.
[26]WARNEKE S,SCHIPPER L A,MATIASEK M G,et al.Nitrate removal,communities of denitrifiers and adverse effects in different carbon substrates for use in denitrification beds[J].Water Research,2011,45(17):5463-5475.
[27]YANG Y,TAO X,LIN E,et al.Enhanced nitrogen removal with spent mushroom compost in a sequencing batch reactor[J].Bioresource Technology,2017,244(Part 1):897.
[28]ZHI W,JI G.Quantitative response relationships between nitrogen transformation rates and nitrogen functional genes in a tidal flow constructed wetland under C/N ratio constraints[J].Water Research,2014,64(7):32-41.
[29]PARANYCHIANAKIS N V,TSIKNIA M,KALOGERAKISN.Pathways regulating the removal of nitrogen in planted and unplanted subsurface flow constructed wetlands[J].Water Research,2016,102:321-329.
[30]FU G P,HUANGSHEN L K,GUO Z P,et al.Effect of plant-based carbon sources on denitrifying microorganisms in a vertical flow constructed wetland[J].Bioresource Technology,2016,224:214-221.
[31]ZHANG X M,HUA X F,YUE X P.Comparison of bacterial community characteristics between complete and shortcut denitrification systems for quinoline degradation[J].Applied Microbiology&Biotechnology,2017,101(4):1697-1707.
[2]李桂平,栾兆坤.深床直接过滤工艺深度处理城市污水[J].中国给水排水,2002,18(10):12-15.LI Guiping,LUAN Zhaokun.Using micro-flocculation deep-bed biofiltration process for advanced treatment of urban wastewater[J].China Water and Wastewater,2002,18(10):12-15.
[3]陈浬,王海,黄韬,等.反硝化滤池深度处理污水厂尾水的效能研究[J].中国给水排水,2017,3(13):99-103.CHEN Li,WANG Hai,HUANG Tao,et al.Treatment efficiency of secondary effluent of WWTP by denitrification filter[J].China Water and Wastewater,2017,3(13):99-103.
[4]沈晓铃,李大成,蒋岚岚,等.深床反硝化滤池在污水厂提标扩建工程中的应用[J].中国给水排水,2010,26(4):32-34.SHEN Xiaoling,LI Dacheng,JIANG Lanlan,et al.Application of deep-bed denitrification filter in upgrading and extension project of WWTP[J].China Water and Wastewater,2010,26(4):32-34.
[5]王庆,丁原红,任洪强,等.采用深床滤池对东港污水厂出水深度脱氮处理[J].工业水处理,2014,34(11):66-68.WANG Qing,DING Yunahong,REN Hongqiang,et al.Deep-fixed bed filtration process used for the advanced denitrification treatment of effluent from Donggang Sewage Treatment Plant[J].Industrial Water Treatment,2014,34(11):66-68.
[6]蒋其宏.单层石英砂滤料改为双层滤料滤池的探索[J].城镇供水,2003(1):9-11.JIANG Qihong.Exploration of changing single-layer quartz sand filter material into double-layer filter material filter[J].City and Town Water Supply,2003(1):9-11.
[7]吴强,陆少鸣.炭砂复合滤池处理微污染水源水的研究[J].中国给水排水,2012,28(15):61-63.WU Qiang,LU Shaoming.GAC/sand filter for treatment of micro-polluted source water[J].China Water and Wastewater,2012,28(15):61-63.
[8]王广智,李伟光,王锐,等.炭砂滤池在松花江污染应急处理中的应用特性研究[J].给水排水,2007,33(8):11-15.WANG Guangzhi,LI Weiguang,WANG Rui,et al.Study on emergent application of combined activated carbon and sand filter in the Songhua River pollution accident[J].Water and Wastewater Engineering,2007,33(8):11-15.
[9]ALTMANN J,RUHL A S,SAUTER D,et al.How to dose powdered activated carbon in deep bed filtration for efficient micropollutant removal[J].Water Research,2015,78:9.
[10]蔡广强,傅学敏,刘丽君,等.曝气炭砂滤池对消毒副产物三氯乙醛的控制作用[J].中国给水排水,2016(5):1-3.CAI Guangqiang,FU Xuemin,LIU Lijun,et al.Control of disinfection by-product chloral hydrate by aerated GAC-sand filter[J].China Water and Wastewater,2016(5):1-3.
[11]张声,刘洋,张晓健,等.活性炭石英砂双层深床滤料浮滤池处理高藻水源水的研究[J].环境科学,2004,25(5):52-56.ZHANG Sheng,LIU Yang,ZHANG Xiaojian,et al.Treating of algae-laden raw water with GAC-sand dual media deep bed dissolved air flotation/filtration[J].Chinese Journal of Environmental Science,2004,25(5):52-56.
[12]张路.炭砂生物滤池工艺在饮用水深度处理中的应用研究[D].北京:北京化工大学,2013.ZHANG Lu.Study on the application of GAC-sand filter in advanced treatment of drinking water[D].Beijing:Beijing University of Chemical Technology,2013.
[13]GUO G X,DENG H,QIAO M,et al.Effect of pyrene on denitrification activity and abundance and composition of denitrifying community in an agricultural soil[J].Environmental Pollution,2011,159(7):1886-1895.
[14]HENRY S,TEXIER S,HALLET S,et al.Disentangling the rhizosphere effect on nitrate reducers and denitrifiers:insight into the role of root exudates[J].Environmental Microbiology,2008,10(11):3082-3092.
[15]BRU D,SARR A,PHILIPPOT L.Relative abundances of proteobacterial membrane-bound and periplasmic nitrate reductases in selected environments[J].Applied&Environmental Microbiology,2007,73(18):5971-5974.
[16]MICHOTEY V,MéJEAN V,BONIN P.Comparison of methods for quantification of cytochrome CD1-denitrifying bacteria in environmental marine samples[J].Applied&Environmental Microbiology,2000,66(4):1564-1571.
[17]HALLIN S,LINDGREN P E.PCR detection of genes encoding nitrite reductase in denitrifying bacteria[J].Applied&Environmental Microbiology,1999,65(4):1652-1657.
[18]HENRY S,BRU D,STRES B,et al.Quantitative detection of the nosZ gene,encoding nitrous oxide reductase,and comparison of the abundances of 16S rRNA,narG,nirK,and nosZ genes in soils[J].Applied&Environmental Microbiology,2006,72(8):5181-5189.
[19]吴兴海,李咏梅.碳氮比对不同滤料反硝化滤池脱氮效果的影响[J].环境工程学报,2017(1):55-62.WU Xinghai,LI Yongmei.Effect of C/N ratio in denitrification of denitrification filters with different filter materials[J].Chinese Journal of Environmental Engineering,2017(1):55-62.
[20]HOU J,XIA L,MA T,et al.Achieving short-cut nitrification and denitrification in modified intermittently aerated constructed wetland[J].Bioresource Technology,2017,232:10.
[21]葛勇涛,焦阳,李军,等.乙酸钠碳源强化生物滤池对二沉池出水的脱氮效果[J].中国给水排水,2011,27(7):98-100.GE Yongtao,JIAO Yang,LI Jun,et al.Sodium acetate as carbon source for enhancing nitrogen removal from secondary effluent with biological filter[J].China Water and Wastewater,2011,27(7):98-100.
[22]HER J J,HUANG J S.Influences of carbon source and C/N ratio on nitrate/nitrite denitrification and carbon breakthrough[J].Bioresource Technology,1995,54(1):45-51.
[23]邹联沛,王宝贞,迟军,等.膜生物反应器处理污水性能的研究[J].哈尔滨建筑大学学报,2001,34(2):57-60.ZOU Lianpei,WANG Baozhen,CHI Jun,et al.Wastewater treating capability of membrane bioreactor[J].Journal of Harbin University of Civil Engineering And Architecture,2001,34(2):57-60.
[24]LEVY-BOOTH D J,PRESCOTT C E,GRAYSTON S J.Microbial functional genes involved in nitrogen fixation,nitrification and denitrification in forest ecosystems[J].Soil Biology&Biochemistry,2014,75:11-25.
[25]张盛博,何小娟,吴海露,等.分子生态学技术用于反硝化微生物群落研究的进展[J].环境科学与技术,2017(6):120-126.ZHANG Shengbo,HE Xiaojuan,WU Hailu,et al.Advances in molecular ecology technologies for the study of denitrifier communities[J].Environmental Science and Technology,2017(6):120-126.
[26]WARNEKE S,SCHIPPER L A,MATIASEK M G,et al.Nitrate removal,communities of denitrifiers and adverse effects in different carbon substrates for use in denitrification beds[J].Water Research,2011,45(17):5463-5475.
[27]YANG Y,TAO X,LIN E,et al.Enhanced nitrogen removal with spent mushroom compost in a sequencing batch reactor[J].Bioresource Technology,2017,244(Part 1):897.
[28]ZHI W,JI G.Quantitative response relationships between nitrogen transformation rates and nitrogen functional genes in a tidal flow constructed wetland under C/N ratio constraints[J].Water Research,2014,64(7):32-41.
[29]PARANYCHIANAKIS N V,TSIKNIA M,KALOGERAKISN.Pathways regulating the removal of nitrogen in planted and unplanted subsurface flow constructed wetlands[J].Water Research,2016,102:321-329.
[30]FU G P,HUANGSHEN L K,GUO Z P,et al.Effect of plant-based carbon sources on denitrifying microorganisms in a vertical flow constructed wetland[J].Bioresource Technology,2016,224:214-221.
[31]ZHANG X M,HUA X F,YUE X P.Comparison of bacterial community characteristics between complete and shortcut denitrification systems for quinoline degradation[J].Applied Microbiology&Biotechnology,2017,101(4):1697-1707.