河道底泥中异养硝化-好氧反硝化菌群富集培养及其脱氮性能
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摘要
采用选择性富集培养基,从河道底泥中成功筛选出具有氮素去除效果的优势微生物菌群:异养硝化-好氧反硝化菌群(H)、异养硝化菌群(Y)和好氧反硝化菌群(F).高通量测序分析结果显示,各混合菌群中脱氮功能菌的相对丰度分别为54.72%、46.26%和91.57%.在此基础上,考察了微量元素对3组混合菌群脱氮性能的影响,结果表明,混合菌群H和F在添加微量元素浓缩液0.5 mL·L~(-1)时脱氮性能最佳,在72 h时混合菌群H对氨氮、硝酸盐氮的去除率分别为69.11%和99.06%,混合菌群F对硝酸盐氮的去除率达99.62%;而混合菌群Y在不添加微量元素时氨氮的去除率最高,为82.60%.采用优化培养的3组混合菌群及其复合菌群(H+Y、Y+F、H+F、H+Y+F)分别对模拟河水进行处理,模拟河水的COD和氮素指标可从地表水V类水质净化至Ⅳ类水质,其中,复合菌群H+Y+F的氮素去除效果最佳,在36 h时氨氮、硝酸盐氮和总氮的浓度分别为0.182、0.000和0.501 mg·L~(-1),说明本研究富集筛选出的混合脱氮菌群在实际河水净化中具有潜在应用价值.
Three cultures, heterotrophic nitrification-aerobic denitrifiers(H), heterotrophic nitrifiers(Y) and aerobic denitrifiers(F), were successfully enriched from river sediment by selective enrichment medium. High-throughput sequencing revealed that the relative abundance of functional bacteria in each community was identified as 54.72%, 46.26% and 91.57%, respectively. trace element effects on nitrogen removal performance for three denitrifying cultures were investigated. The results show that culture H and F exhibited the best nitrogen removal performance at 0.5 mL·L~(-1) of trace elements. The removal efficiencies of ammonia and nitrate by culture H were 69.11% and 99.65% after 72 hour, and the removal efficiencies of nitrate by culture F reached 99.62%. However, the removal efficiency of ammonia was the highest at 82.60% for culture Y when no trace elements were added at the 72 hour. The COD and nitrogen indexes of the simulated river water could be purified from class V of quality standards for surface water to class IV after these three optimized single and their mixed cultures(H+Y, Y+F, H+F, H+Y+F) inoculated respectively. The mixed culture(H+Y+F) had the highest nitrogen removal efficiencies, with 0.182 mg·L~(-1) ammonia, 0.000 mg·L~(-1) nitrate and 0.501 mg·L~(-1) total nitrogen(TN) of the simulated river water at the 36 hour. The selected three denitrifying culture have potential application value in actual river water purification.
Three cultures, heterotrophic nitrification-aerobic denitrifiers(H), heterotrophic nitrifiers(Y) and aerobic denitrifiers(F), were successfully enriched from river sediment by selective enrichment medium. High-throughput sequencing revealed that the relative abundance of functional bacteria in each community was identified as 54.72%, 46.26% and 91.57%, respectively. trace element effects on nitrogen removal performance for three denitrifying cultures were investigated. The results show that culture H and F exhibited the best nitrogen removal performance at 0.5 mL·L~(-1) of trace elements. The removal efficiencies of ammonia and nitrate by culture H were 69.11% and 99.65% after 72 hour, and the removal efficiencies of nitrate by culture F reached 99.62%. However, the removal efficiency of ammonia was the highest at 82.60% for culture Y when no trace elements were added at the 72 hour. The COD and nitrogen indexes of the simulated river water could be purified from class V of quality standards for surface water to class IV after these three optimized single and their mixed cultures(H+Y, Y+F, H+F, H+Y+F) inoculated respectively. The mixed culture(H+Y+F) had the highest nitrogen removal efficiencies, with 0.182 mg·L~(-1) ammonia, 0.000 mg·L~(-1) nitrate and 0.501 mg·L~(-1) total nitrogen(TN) of the simulated river water at the 36 hour. The selected three denitrifying culture have potential application value in actual river water purification.
引文
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康鹏亮,张海涵,黄廷林,等.2018.湖库沉积物好氧反硝化菌群脱氮特性及种群结构[J].环境科学,39(5):2431-2437
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Li A,Gai Z,Cui D,et al.2012.Genome sequence of a highly efficient aerobic denitrifying bacterium,Pseudomonas stutzeri T13[J].Journal of Bacteriology,194(20):5720-5720
Li C,Yang J,Wang X,et al.2015.Removal of nitrogen by heterotrophic nitrification–aerobic denitrification of a phosphate accumulating bacterium Pseudomonas stutzeri YG-24[J].Bioresource Technology,182:18-25
Liu Y,Wang Y,Li Y,et al.2015.Nitrogen removal characteristics of heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis C16[J].Chinese Journal of Chemical Engineering,23(5):827-834
刘尚俭,邱建,安雅敏,等.2015.河流生态修复浅议[J].环境影响评价,37(1):82-85
Medhi K,Singhal A,Chauhan D K,et al.2017.Investigating the nitrification and denitrification kinetics under aerobic and anaerobic conditions by Paracoccus denitrificans ISTOD1[J].Bioresource Technology,242:334-343
Perelo L W.2010.Review:In situ and bioremediation of organic pollutants in aquatic sediments[J].Journal of Hazardous Materials,177(1/3):81-89
Richardson D J,Wehrfritz J M,Keech A,et al.1998.The diversity of redox proteins involved in bacterial heterotrophic nitrification and aerobic denitrification[J].Biochemical Society Transactions,26(3):401-408
Schmidt I,Sliekers O,Schmid M,et al.2003.New concepts of microbial treatment process for the nitrogen removal in wastewater[J].Fems Microbiology Reviews,27(4):481-492
Su J F,Zhang K,Huang T L,et al.2015.Heterotrophic nitrification and aerobic denitrification at low nutrient conditions by a newly isolated bacterium,Acinetobacter sp.SYF26[J].Microbiology,161(4):829-837
史佳媛.2015.脱氮优势菌群筛选及其固定化应用于河道底泥修复[D].南京:东南大学.1-61
宋琴,许雷.2008.异养硝化作用酶学研究进展[J].生物技术通报,(5):60-62
宋钊.2013.城市河流水污染治理及修复技术[J].工业用水与废水,44(4):6-8
苏俊峰,王继华,马放,等.2007.好氧反硝化细菌的筛选鉴定及处理硝酸盐废水的研究[J].环境科学,28(10):2332-2335
孙菲,方金武,蔡昌凤,等.2010.好氧反硝化菌群的筛选及其培养条件的研究[J].安徽工程大学学报,25(2):36-39
Wan W,He D,Xue Z.2017.Removal of nitrogen and phosphorus by heterotrophic nitrification-aerobic denitrification of a denitrifying phosphorus-accumulating bacterium Enterobacter cloacae HW-15[J].Ecological Engineering,99:199-208
王景峰,金敏,谌志强.2011.1株好氧脱氮菌的筛选与脱氮特性研究[J].环境科学,32(8):2409-2413
王田野,魏荷芬,胡子全,等.2017.一株异养硝化好氧反硝化菌的筛选鉴定及其脱氮特性[J].环境科学学报,37(3):945-952
王瑶,刘玉香,安华,等.2014.金属离子对粪产碱杆菌C16的脱氮和亚硝酸盐积累的影响[J].微生物学通报,41(11):2254-2263
王永刚,王旭,张俊娥,等.2017.好氧反硝化细菌研究及应用进展[J].工业水处理,(2):12-17
杨婷,杨娅,刘玉香.2017.异养硝化-好氧反硝化的研究进展[J].微生物学通报,44(9):2213-2222
尹明锐,汪苹,刘健楠,等.2010.复合脱氮菌群的构建及其脱氮特性研究[J].食品科学技术学报,28(3):55-59
雍佳君,成小英.2015.蠡河底泥中反硝化复合菌群富集及菌群结构研究[J].环境科学,36(6):2232-2238
于大禹,张琳颖,高波.2012.异养硝化-好氧反硝化菌异养硝化性能的影响因素[J].化工进展,31(12):2797-2800
Yang L,Ren Y X,Liang X,et al.2015.Nitrogen removal characteristics of a heterotrophic nitrifier Acinetobacter junii YB and its potential application for the treatment of high-strength nitrogenous wastewater[J].Bioresource Technology,193:227-233
Yang X P,Wang S M,Zhang D W,et al.2011.Isolation and nitrogen removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium,Bacillus subtilis A1[J].Bioresource Technology,102(2):854-862
Zhang S,Sun X,Fan Y,et al.2017.Heterotrophic nitrification and aerobic denitrification by Diaphorobacter polyhydroxybutyrativorans SL-205 using poly(3-hydroxybutyrate-co-3-hydro-xyvalerate) as the sole carbon source[J].Bioresource Technology,241:500-507
Zhao B,Cheng D Y,Tan P,et al.2018.Characterization of an aerobic denitrifier Pseudomonas stutzeri strain XL-2 to achieve efficient nitrate removal[J].Bioresource Technology,250:564-573
Zhao B,He Y L,Huang J,et al.2010.Heterotrophic nitrogen removal by Providencia rettgeri strain YL[J].Journal of Industrial Microbiology and Biotechnology,37(6):609-616
Zhao B,He Y,Hughes J,et al.2010.Heterotrophic nitrogen removal by a newly isolated Acinetobacter calcoaceticus HNR[J].Bioresource Technology,101(14):5194-5200
郭琳.2015.水源水库中好氧反硝化菌的筛选及脱氮性能研究[D].西安:西安建筑科技大学.1-131
国家环境保护总局.2002.水和废水监测分析方法(第4版)[M].北京:中国环境科学出版社.1-784
He T,Li Z,Sun Q,et al.2016.Heterotrophic nitrification and aerobic denitrification by Pseudomonas tolaasii Y-11 without nitrite accumulation during nitrogen conversion[J].Bioresource Technology,200(1):493-499
何霞,吕剑,何义亮,等.2006.异养硝化机理的研究进展[J].微生物学报,46(5):844-847
环境保护部.2017.2017年中国生态环境状况公报[R].北京:环境保护部.1-55
黄郑郑,曹刚,陈海升,等.2017.异养硝化-好氧反硝化菌协同竞争对脱氮特性的影响[J].环境科学研究,30(5):765-7
康鹏亮,张海涵,黄廷林,等.2018.湖库沉积物好氧反硝化菌群脱氮特性及种群结构[J].环境科学,39(5):2431-2437
Kim J K,Park K J,Cho K S,et al.2005.Aerobic nitrification-denitrification by heterotrophic Bacillus strains[J].Bioresource Technology,96(17):1897-1906
Li A,Gai Z,Cui D,et al.2012.Genome sequence of a highly efficient aerobic denitrifying bacterium,Pseudomonas stutzeri T13[J].Journal of Bacteriology,194(20):5720-5720
Li C,Yang J,Wang X,et al.2015.Removal of nitrogen by heterotrophic nitrification–aerobic denitrification of a phosphate accumulating bacterium Pseudomonas stutzeri YG-24[J].Bioresource Technology,182:18-25
Liu Y,Wang Y,Li Y,et al.2015.Nitrogen removal characteristics of heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis C16[J].Chinese Journal of Chemical Engineering,23(5):827-834
刘尚俭,邱建,安雅敏,等.2015.河流生态修复浅议[J].环境影响评价,37(1):82-85
Medhi K,Singhal A,Chauhan D K,et al.2017.Investigating the nitrification and denitrification kinetics under aerobic and anaerobic conditions by Paracoccus denitrificans ISTOD1[J].Bioresource Technology,242:334-343
Perelo L W.2010.Review:In situ and bioremediation of organic pollutants in aquatic sediments[J].Journal of Hazardous Materials,177(1/3):81-89
Richardson D J,Wehrfritz J M,Keech A,et al.1998.The diversity of redox proteins involved in bacterial heterotrophic nitrification and aerobic denitrification[J].Biochemical Society Transactions,26(3):401-408
Schmidt I,Sliekers O,Schmid M,et al.2003.New concepts of microbial treatment process for the nitrogen removal in wastewater[J].Fems Microbiology Reviews,27(4):481-492
Su J F,Zhang K,Huang T L,et al.2015.Heterotrophic nitrification and aerobic denitrification at low nutrient conditions by a newly isolated bacterium,Acinetobacter sp.SYF26[J].Microbiology,161(4):829-837
史佳媛.2015.脱氮优势菌群筛选及其固定化应用于河道底泥修复[D].南京:东南大学.1-61
宋琴,许雷.2008.异养硝化作用酶学研究进展[J].生物技术通报,(5):60-62
宋钊.2013.城市河流水污染治理及修复技术[J].工业用水与废水,44(4):6-8
苏俊峰,王继华,马放,等.2007.好氧反硝化细菌的筛选鉴定及处理硝酸盐废水的研究[J].环境科学,28(10):2332-2335
孙菲,方金武,蔡昌凤,等.2010.好氧反硝化菌群的筛选及其培养条件的研究[J].安徽工程大学学报,25(2):36-39
Wan W,He D,Xue Z.2017.Removal of nitrogen and phosphorus by heterotrophic nitrification-aerobic denitrification of a denitrifying phosphorus-accumulating bacterium Enterobacter cloacae HW-15[J].Ecological Engineering,99:199-208
王景峰,金敏,谌志强.2011.1株好氧脱氮菌的筛选与脱氮特性研究[J].环境科学,32(8):2409-2413
王田野,魏荷芬,胡子全,等.2017.一株异养硝化好氧反硝化菌的筛选鉴定及其脱氮特性[J].环境科学学报,37(3):945-952
王瑶,刘玉香,安华,等.2014.金属离子对粪产碱杆菌C16的脱氮和亚硝酸盐积累的影响[J].微生物学通报,41(11):2254-2263
王永刚,王旭,张俊娥,等.2017.好氧反硝化细菌研究及应用进展[J].工业水处理,(2):12-17
杨婷,杨娅,刘玉香.2017.异养硝化-好氧反硝化的研究进展[J].微生物学通报,44(9):2213-2222
尹明锐,汪苹,刘健楠,等.2010.复合脱氮菌群的构建及其脱氮特性研究[J].食品科学技术学报,28(3):55-59
雍佳君,成小英.2015.蠡河底泥中反硝化复合菌群富集及菌群结构研究[J].环境科学,36(6):2232-2238
于大禹,张琳颖,高波.2012.异养硝化-好氧反硝化菌异养硝化性能的影响因素[J].化工进展,31(12):2797-2800
Yang L,Ren Y X,Liang X,et al.2015.Nitrogen removal characteristics of a heterotrophic nitrifier Acinetobacter junii YB and its potential application for the treatment of high-strength nitrogenous wastewater[J].Bioresource Technology,193:227-233
Yang X P,Wang S M,Zhang D W,et al.2011.Isolation and nitrogen removal characteristics of an aerobic heterotrophic nitrifying-denitrifying bacterium,Bacillus subtilis A1[J].Bioresource Technology,102(2):854-862
Zhang S,Sun X,Fan Y,et al.2017.Heterotrophic nitrification and aerobic denitrification by Diaphorobacter polyhydroxybutyrativorans SL-205 using poly(3-hydroxybutyrate-co-3-hydro-xyvalerate) as the sole carbon source[J].Bioresource Technology,241:500-507
Zhao B,Cheng D Y,Tan P,et al.2018.Characterization of an aerobic denitrifier Pseudomonas stutzeri strain XL-2 to achieve efficient nitrate removal[J].Bioresource Technology,250:564-573
Zhao B,He Y L,Huang J,et al.2010.Heterotrophic nitrogen removal by Providencia rettgeri strain YL[J].Journal of Industrial Microbiology and Biotechnology,37(6):609-616
Zhao B,He Y,Hughes J,et al.2010.Heterotrophic nitrogen removal by a newly isolated Acinetobacter calcoaceticus HNR[J].Bioresource Technology,101(14):5194-5200
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