脱氮微生物制剂的研制
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摘要
针对我国现有多数污水处理设施只去除有机物和固体悬浮物，不具有脱氮功能，我们在试验中开发了一种简捷、高效的脱氮微生物制剂，将其直接投入处理设施的曝气池中，使得能够实现脱氮的目的。
脱氮微生物制剂的制备是将亚硝酸菌、反硝化菌的高密度培养液高速离心，菌体浓缩物进行适当配比，用PVA载体固定而成。试验研究了亚硝酸菌和反硝化菌的高密度培养条件、用PVA固定脱氮菌的方法、实现高效脱氮的环境条件、脱氮微生物制剂处理实际废水的效果。试验结果表明：
一、用硅胶平板分离亚硝酸细菌的富集培养液，分离得到亚硝化球菌和亚硝化单胞菌，对分离到的这两株菌扩大培养六周后，进一步组合筛选，得到氨氧化速率高的混合菌株，应用正交试验法对其高密度培养条件进行试验，得到高密度培养的最适条件：NaHCO_3为lg/L，(NH_4)_2SO_4为2g/L，pH值8.0，微量元素为1ml/L及溶解氧大于2.4mg/L。在此条件下，亚硝酸细菌的高密度培养液菌体浓度达到1.4×10~9MPN/ml。
二、在亚硝化型反硝化菌体富集培养液中，筛选出三株脱氮率高的菌株。这三株菌体的等比混合物为脱氮率最高的种型，研究结果表明：采用乙酸钠为碳源和能源富集培养反硝化细菌比采用甲醇、乙醇或酒石酸钾钠更易分离筛选出高效生物脱氮菌种；试验菌种在脱氮过程中没有硝酸盐的生成；反硝化菌的最适高密度培养条件为接种量10％，C/N 2.5，pH值7.0～8.0之间，温度30℃。
三、采用聚乙烯醇(PVA)-硼酸包埋固定化法，选用PVA为包埋载体，粉末活性炭作为无机载体，包埋固定实验分离得到的亚硝酸菌和反硝化菌，制成固定化脱氮微生物颗粒。试验结果表明：当COD处于500～1000 mg/L内，pH值在8.0～8.5之间，温度和溶解氧分别是30℃和2～6mg/L，此时，NH_4-N去除率达90％以上。
四、以流化床作为生物反应器，采用SBR运行方式对实际城市生活污水进行处理试验。当颗粒填充率为10％，进水NH_4-N浓度为40mg/L，反应时间为8h时，氨氮去除率可达90％以上，TN的去除率为80％左右，同时COD的去除率约40％。
Most traditional effluent disposal establishments only be able to removal organic pollutants and solid suspend but have not the function of nitrogen removal. As to the problems, a kind of convenient and efficient microbial preparation for nitrogen removal was developed in this paper. As it was directly cast into the aeration tank, the effluent disposal establishments is capable of the function of nitrogen removal.
The microbial preparation for nitrogen removal was equipped by making the high density culture liquid of nitrous bacteria and denitrifying bacteria centrifugal, mixing condensate bacteria in appropriate proportion and immobilizing the mixture with polyvinyl alcohol (PVA). In this thesis, the condition of culturing the nitrous bacteria and denitrifying bacteria of high density was determined, the method of immobilization by PVA was established, the environment for realizing nitrogen removal at high speed was studied, and the result of using the microbial preparation for nitrogen removal to treat waste water was detected
On the silicate plate, pure nitrobacteria were obtained. Strain Y1 was identified to be Nitrisospira sp. Strain Y2 was Nitrosococcua sp. By extending the two bacteria for 6 weeks and selecting further, strain mixed of high amide oxidation rate produced. The results showed that under the optimum condition: NaHCO3 1g/L, (NH4) 2SO4 2g/L, pH 8.5, DO>2.4mg/L, trace element solution 1 ml, the concentration of nitrobacteria was up to 1.4×109MPN /ml
Three high denitrification bacteria were isolated from sludge enrichment culture. The mixture of the three strains in same share is the type of highest nitrogen removal rate. The results showed that the high affection removal nitrogen bacteria screened out by sodium acetate was more easily than using carbinol, ethanol, Kalium natriumtartaric acid as carbon and energy sources. And nitrate wasn't produced in the course of nitrogen removal. The optimum condition for high-density cultures of denitrification bacteria was inoculation 10%, C/N 2.5, pH 7.0 to 8.0 and temperature 30℃.
The immobilized microbes pellets were made from nitrosobacteria and denitrifying bacteria using polyvinyl alcohol(PVA)cross linked with boric acid ,PVA is embed carrier and powdered activated carbon is inorganic carrier. The source of nitrosobacteria and denitrifying bacteria come from activated sludge system. The results showed that while the COD between 500 and 1000mg/L,pH8.0 to 8.5 ,temperature 30℃ and DO 2 to 6mg/L,the removal efficiency -reached 90%.
In biological fluidized bed reactor, Sequencing Batch Reactors system treating practical life wastewater was carried out. While the immobilized microbe's pellets package percentage was 10%, influent NH4-N concentration was 40 mg/L, reaction time was 8h, the NH4-N removal efficiency reach 90%, the TN removal efficiency could reach 80%; the COD removal efficiency could reach 40% as well.
脱氮微生物制剂的制备是将亚硝酸菌、反硝化菌的高密度培养液高速离心，菌体浓缩物进行适当配比，用PVA载体固定而成。试验研究了亚硝酸菌和反硝化菌的高密度培养条件、用PVA固定脱氮菌的方法、实现高效脱氮的环境条件、脱氮微生物制剂处理实际废水的效果。试验结果表明：
一、用硅胶平板分离亚硝酸细菌的富集培养液，分离得到亚硝化球菌和亚硝化单胞菌，对分离到的这两株菌扩大培养六周后，进一步组合筛选，得到氨氧化速率高的混合菌株，应用正交试验法对其高密度培养条件进行试验，得到高密度培养的最适条件：NaHCO_3为lg/L，(NH_4)_2SO_4为2g/L，pH值8.0，微量元素为1ml/L及溶解氧大于2.4mg/L。在此条件下，亚硝酸细菌的高密度培养液菌体浓度达到1.4×10~9MPN/ml。
二、在亚硝化型反硝化菌体富集培养液中，筛选出三株脱氮率高的菌株。这三株菌体的等比混合物为脱氮率最高的种型，研究结果表明：采用乙酸钠为碳源和能源富集培养反硝化细菌比采用甲醇、乙醇或酒石酸钾钠更易分离筛选出高效生物脱氮菌种；试验菌种在脱氮过程中没有硝酸盐的生成；反硝化菌的最适高密度培养条件为接种量10％，C/N 2.5，pH值7.0～8.0之间，温度30℃。
三、采用聚乙烯醇(PVA)-硼酸包埋固定化法，选用PVA为包埋载体，粉末活性炭作为无机载体，包埋固定实验分离得到的亚硝酸菌和反硝化菌，制成固定化脱氮微生物颗粒。试验结果表明：当COD处于500～1000 mg/L内，pH值在8.0～8.5之间，温度和溶解氧分别是30℃和2～6mg/L，此时，NH_4-N去除率达90％以上。
四、以流化床作为生物反应器，采用SBR运行方式对实际城市生活污水进行处理试验。当颗粒填充率为10％，进水NH_4-N浓度为40mg/L，反应时间为8h时，氨氮去除率可达90％以上，TN的去除率为80％左右，同时COD的去除率约40％。
Most traditional effluent disposal establishments only be able to removal organic pollutants and solid suspend but have not the function of nitrogen removal. As to the problems, a kind of convenient and efficient microbial preparation for nitrogen removal was developed in this paper. As it was directly cast into the aeration tank, the effluent disposal establishments is capable of the function of nitrogen removal.
The microbial preparation for nitrogen removal was equipped by making the high density culture liquid of nitrous bacteria and denitrifying bacteria centrifugal, mixing condensate bacteria in appropriate proportion and immobilizing the mixture with polyvinyl alcohol (PVA). In this thesis, the condition of culturing the nitrous bacteria and denitrifying bacteria of high density was determined, the method of immobilization by PVA was established, the environment for realizing nitrogen removal at high speed was studied, and the result of using the microbial preparation for nitrogen removal to treat waste water was detected
On the silicate plate, pure nitrobacteria were obtained. Strain Y1 was identified to be Nitrisospira sp. Strain Y2 was Nitrosococcua sp. By extending the two bacteria for 6 weeks and selecting further, strain mixed of high amide oxidation rate produced. The results showed that under the optimum condition: NaHCO3 1g/L, (NH4) 2SO4 2g/L, pH 8.5, DO>2.4mg/L, trace element solution 1 ml, the concentration of nitrobacteria was up to 1.4×109MPN /ml
Three high denitrification bacteria were isolated from sludge enrichment culture. The mixture of the three strains in same share is the type of highest nitrogen removal rate. The results showed that the high affection removal nitrogen bacteria screened out by sodium acetate was more easily than using carbinol, ethanol, Kalium natriumtartaric acid as carbon and energy sources. And nitrate wasn't produced in the course of nitrogen removal. The optimum condition for high-density cultures of denitrification bacteria was inoculation 10%, C/N 2.5, pH 7.0 to 8.0 and temperature 30℃.
The immobilized microbes pellets were made from nitrosobacteria and denitrifying bacteria using polyvinyl alcohol(PVA)cross linked with boric acid ,PVA is embed carrier and powdered activated carbon is inorganic carrier. The source of nitrosobacteria and denitrifying bacteria come from activated sludge system. The results showed that while the COD between 500 and 1000mg/L,pH8.0 to 8.5 ,temperature 30℃ and DO 2 to 6mg/L,the removal efficiency -reached 90%.
In biological fluidized bed reactor, Sequencing Batch Reactors system treating practical life wastewater was carried out. While the immobilized microbe's pellets package percentage was 10%, influent NH4-N concentration was 40 mg/L, reaction time was 8h, the NH4-N removal efficiency reach 90%, the TN removal efficiency could reach 80%; the COD removal efficiency could reach 40% as well.
引文
[1]刘雨，赵庆良，郑兴灿．生物膜法污水处理技术(第一版)．北京：中国建筑工业出版社，2000：45
[2]徐亚同编著．废水中氮磷的处理．上海：华东师范大学出版社，1996
[3]Yuicbi Suwa, and Yasuo Imamura, et al. Ammonia-oxidizing bacteria with different sensitivities to (NH_4)_2SO_4in activated sledges. Wat Res, 1994,28(7):1523～1532.
[4]章非娟编著．生物脱氮技术．第一版，北京：中国环境科学出版社，1993，17．
[5]Tanaka K, Sumina T, Nakamura H et al. Application of nitrification by cells immobilized in polyethylene glycoi. Prog. Biotechnol. 1996,11(immobilized cells):622～632
[6]Mishima, koji; Watanabe, Akira; endo, kaneaki. Immobilization of enzymes or microorganisms Jpn. Kokai Tokkyo Koho JP1216688[86216688](IPC C12N011/88), 26 Sep 1986, Appl. 85/54383,20Mar 1985:5
[7]Nilssion I Eur. J. Appli. Microbiol. Biotechnol, 1980:10:261
[8]Tsai, Yu Li; Schlasner, Steven M. Tuovineo, Olli H. Inhibitor evaluation with immobilized Nitrobacteria argils cells. Appl. Environ. Microbial. 1986;. 52(6):12311235
[9]Mach Ida, Nobuyki; Kimura, Jun Biochemical immobilization using spherical polyacrylamide gel. Jpn. Kokai Tokkyo Koho JP1216688186216688](IPC C12NO11/88),2Feb 1988, Appl. 86/170247,18jul 1986:5
[10]中村．固定化生物脱氮研究．公害与对策，1987，22(2)：27～30
[11]Konishi, Kozo; Kobayashi, Minoru. Identification method and apparatus for ammonium removal from wastewater. Jpn. Kokai Tokkyo Koho JP 62258795[87258795] (ipccO2F 003/06),11Nov1987, Appl.86/80184,9Apr1986:4
[12]Menzel, Roman; Hoffmann, Peter; Vorlop, Removal of ammonium from wastewater, surface waters, or ground-water Ger. Offen. DE 3641567 (ipcc02f003/02), 9Jun 1988, Appl. 3641567, 5Dec 1988:5Addn. to Ger. Offen. 3, 520,160.
[13]Kokufuta, Etsuo; Yuishige, Masaya; Nakamura, Isei. Co immobilization of Nitrosomonas europea and Paracoccus denitrificans cells using poluelectroly te complex-stabilized calcium alginate gel. J. Ferment. Technol. 1987;65(6)659-664
[14]王家玲主编．环境微生物学实验北京：高等教育出版社，1988：40～44
[15]金志刚，屈计宁．硝化细菌富集技术分析及方法研究．上海环境科学，1998，8：16～19
[16]国家环保局《水和废水监测分析方法》编委会编。水和废水监测分析方法(第三版)．北京：中国环境科学出版社，1989．254～354
[17]R．E．布坎南，N．E．吉本斯等编，伯杰细菌鉴定手册．(中国科学院微生物研究所《伯杰细菌鉴定手册》翻译组译，北京：科学出版社，1984：622-630
[18]Anthonisen, A.C. et al. Inhibition of nitrification by ammonia and nitrous acid. Water Pollut Control, 197648(2):935～852.
[19]陈金声，史家梁，徐亚同．硝化速率测定和硝化细菌计数考察脱氮效果的应用[J]．上海环境科学，1996，15(3)：18～20．
[20](日)宗宫功编著，张楠等译．污水除磷脱氮技术．第一版，北京：中国环境科学出版社，1987，132
[21]A haron Abeliovich. Transformations of ammonia and the ervironmental impact of nitrifying bacteria. Biodegradation, 1992,21(3):255～264.
[22]黄霞．固定化细胞技术在废水中的应用．环境科学，1993，14(1)：41
[23]赵利兴，兰淑澄．固定化硝化细菌去除废水中氨氮工艺的研究．环境科学．1999，20(1)：39
[24]liujn F.V, Boers P.C, et al. Nitrogen fluxes and process in sandy and muddy sediments from a shallow eutrophic lake.Wat. Res. 1999,33(1):33～42
[25]张蔚文，张灼．混合菌培养在生物降解中的意义．环境科学与技术，1993．(1)：16～20
[26]周康群，黄灿东．降解水源氨氮的高效菌株及组合筛选．环境污染与防治，2001，23(5)：222～224
[27]谭佑铭，罗启芳．固定化反硝化菌对富营养化水体脱氮的试验研究．中国卫生工程学．2003，2(2)：65～68
[28]曹国民，赵庆祥．固定化微生物在好氧条件下同时硝化和反硝化．环境工程，2000，18(5)：17～19
[29]吴晓磊等．海藻酸钠和聚乙烯醇作为固定化微生物包埋剂的研究．环境科学，1992，14(2)：28～31
[30]刘连成．中国湖泊富营养化的现状分析．灾害学，1997．12(3)：61～65
[31]袁林江．聚乙烯醇固定化反硝化菌的脱氮试验研究．西安建筑科技大学学报，1997，29(3)：257～261
[32]安立超，孙静等．混合固定细菌单级生物脱氮技术．污染防治技术，2003，16(3)：5～7
[33]J X Liu, W G Li, et al. Removal of Nitrogen from coal gasification wastewater by nitrosofication and denitrosofication[J].Water Science and Technology, 1998,38(1):39～46
[34]唐光临，孙国新等．焦化废水的亚硝化反硝化研究．水处理技术，2002，28(2)：98～100
[35]使林江，彭党聪．短程硝化-反硝化生物脱氮．中国给水排水，2000，16(2)：29～31
[36]刘俊新，李伟光．亚硝酸型硝化-反硝化工艺处理煤气废水研究．中国给水排
水，1998，14(1)：6～8
[37]李正魁，濮培民．净化湖泊水体氮污染的固定化硝化-反硝化菌研究．湖泊科学，2000，12(2)：119～123
[38]Louis AS, Maja VVA. Five years of nitrate removal, denitrification and carbon dynamics in a denitrification wall. War Res, 2001, 35(14):3473～3477
[39]郑兴灿，李亚新．污水除磷脱氮技术，中国建筑工业出版社，1998．11
[40]Lazarova V, and Mannem J. Innovative process for waste treatment:The circulating Fliating Bed reactor, Wat. Sci.&Tech, 1996,34(9):89～99
[41]Lazarova V, Nogueira r, and Mannem J. Control of nitrification affiance in a new biofilm reactor, Wat. Sci.&Tech, t997,36(1):31～41
[42]Randall CW, Bath D. Nitrite build-up in active sludge resulting from temperature effects, JWPCF, 1984,56(9):1039～1044
[43]Sharma B. Ahlert rc. Nitrification and nitrogen removal. Water Res, 1997,11(9):897～925
[44]刘俊新，王秀蘅．高浓度氨氮废水亚硝酸型与硝酸型脱氮的比较研究．工业用水与废水，2002，33(3)：1～4
[45]Uemoto H, saiki H. Behavior of immobilized nitrosomonas European and paracoccus denitrificans in tubular gel for nitrogen removal in waste water. Pro. Biotechnol, 1996,11(immobilized cells): 695～701
[2]徐亚同编著．废水中氮磷的处理．上海：华东师范大学出版社，1996
[3]Yuicbi Suwa, and Yasuo Imamura, et al. Ammonia-oxidizing bacteria with different sensitivities to (NH_4)_2SO_4in activated sledges. Wat Res, 1994,28(7):1523～1532.
[4]章非娟编著．生物脱氮技术．第一版，北京：中国环境科学出版社，1993，17．
[5]Tanaka K, Sumina T, Nakamura H et al. Application of nitrification by cells immobilized in polyethylene glycoi. Prog. Biotechnol. 1996,11(immobilized cells):622～632
[6]Mishima, koji; Watanabe, Akira; endo, kaneaki. Immobilization of enzymes or microorganisms Jpn. Kokai Tokkyo Koho JP1216688[86216688](IPC C12N011/88), 26 Sep 1986, Appl. 85/54383,20Mar 1985:5
[7]Nilssion I Eur. J. Appli. Microbiol. Biotechnol, 1980:10:261
[8]Tsai, Yu Li; Schlasner, Steven M. Tuovineo, Olli H. Inhibitor evaluation with immobilized Nitrobacteria argils cells. Appl. Environ. Microbial. 1986;. 52(6):12311235
[9]Mach Ida, Nobuyki; Kimura, Jun Biochemical immobilization using spherical polyacrylamide gel. Jpn. Kokai Tokkyo Koho JP1216688186216688](IPC C12NO11/88),2Feb 1988, Appl. 86/170247,18jul 1986:5
[10]中村．固定化生物脱氮研究．公害与对策，1987，22(2)：27～30
[11]Konishi, Kozo; Kobayashi, Minoru. Identification method and apparatus for ammonium removal from wastewater. Jpn. Kokai Tokkyo Koho JP 62258795[87258795] (ipccO2F 003/06),11Nov1987, Appl.86/80184,9Apr1986:4
[12]Menzel, Roman; Hoffmann, Peter; Vorlop, Removal of ammonium from wastewater, surface waters, or ground-water Ger. Offen. DE 3641567 (ipcc02f003/02), 9Jun 1988, Appl. 3641567, 5Dec 1988:5Addn. to Ger. Offen. 3, 520,160.
[13]Kokufuta, Etsuo; Yuishige, Masaya; Nakamura, Isei. Co immobilization of Nitrosomonas europea and Paracoccus denitrificans cells using poluelectroly te complex-stabilized calcium alginate gel. J. Ferment. Technol. 1987;65(6)659-664
[14]王家玲主编．环境微生物学实验北京：高等教育出版社，1988：40～44
[15]金志刚，屈计宁．硝化细菌富集技术分析及方法研究．上海环境科学，1998，8：16～19
[16]国家环保局《水和废水监测分析方法》编委会编。水和废水监测分析方法(第三版)．北京：中国环境科学出版社，1989．254～354
[17]R．E．布坎南，N．E．吉本斯等编，伯杰细菌鉴定手册．(中国科学院微生物研究所《伯杰细菌鉴定手册》翻译组译，北京：科学出版社，1984：622-630
[18]Anthonisen, A.C. et al. Inhibition of nitrification by ammonia and nitrous acid. Water Pollut Control, 197648(2):935～852.
[19]陈金声，史家梁，徐亚同．硝化速率测定和硝化细菌计数考察脱氮效果的应用[J]．上海环境科学，1996，15(3)：18～20．
[20](日)宗宫功编著，张楠等译．污水除磷脱氮技术．第一版，北京：中国环境科学出版社，1987，132
[21]A haron Abeliovich. Transformations of ammonia and the ervironmental impact of nitrifying bacteria. Biodegradation, 1992,21(3):255～264.
[22]黄霞．固定化细胞技术在废水中的应用．环境科学，1993，14(1)：41
[23]赵利兴，兰淑澄．固定化硝化细菌去除废水中氨氮工艺的研究．环境科学．1999，20(1)：39
[24]liujn F.V, Boers P.C, et al. Nitrogen fluxes and process in sandy and muddy sediments from a shallow eutrophic lake.Wat. Res. 1999,33(1):33～42
[25]张蔚文，张灼．混合菌培养在生物降解中的意义．环境科学与技术，1993．(1)：16～20
[26]周康群，黄灿东．降解水源氨氮的高效菌株及组合筛选．环境污染与防治，2001，23(5)：222～224
[27]谭佑铭，罗启芳．固定化反硝化菌对富营养化水体脱氮的试验研究．中国卫生工程学．2003，2(2)：65～68
[28]曹国民，赵庆祥．固定化微生物在好氧条件下同时硝化和反硝化．环境工程，2000，18(5)：17～19
[29]吴晓磊等．海藻酸钠和聚乙烯醇作为固定化微生物包埋剂的研究．环境科学，1992，14(2)：28～31
[30]刘连成．中国湖泊富营养化的现状分析．灾害学，1997．12(3)：61～65
[31]袁林江．聚乙烯醇固定化反硝化菌的脱氮试验研究．西安建筑科技大学学报，1997，29(3)：257～261
[32]安立超，孙静等．混合固定细菌单级生物脱氮技术．污染防治技术，2003，16(3)：5～7
[33]J X Liu, W G Li, et al. Removal of Nitrogen from coal gasification wastewater by nitrosofication and denitrosofication[J].Water Science and Technology, 1998,38(1):39～46
[34]唐光临，孙国新等．焦化废水的亚硝化反硝化研究．水处理技术，2002，28(2)：98～100
[35]使林江，彭党聪．短程硝化-反硝化生物脱氮．中国给水排水，2000，16(2)：29～31
[36]刘俊新，李伟光．亚硝酸型硝化-反硝化工艺处理煤气废水研究．中国给水排
水，1998，14(1)：6～8
[37]李正魁，濮培民．净化湖泊水体氮污染的固定化硝化-反硝化菌研究．湖泊科学，2000，12(2)：119～123
[38]Louis AS, Maja VVA. Five years of nitrate removal, denitrification and carbon dynamics in a denitrification wall. War Res, 2001, 35(14):3473～3477
[39]郑兴灿，李亚新．污水除磷脱氮技术，中国建筑工业出版社，1998．11
[40]Lazarova V, and Mannem J. Innovative process for waste treatment:The circulating Fliating Bed reactor, Wat. Sci.&Tech, 1996,34(9):89～99
[41]Lazarova V, Nogueira r, and Mannem J. Control of nitrification affiance in a new biofilm reactor, Wat. Sci.&Tech, t997,36(1):31～41
[42]Randall CW, Bath D. Nitrite build-up in active sludge resulting from temperature effects, JWPCF, 1984,56(9):1039～1044
[43]Sharma B. Ahlert rc. Nitrification and nitrogen removal. Water Res, 1997,11(9):897～925
[44]刘俊新，王秀蘅．高浓度氨氮废水亚硝酸型与硝酸型脱氮的比较研究．工业用水与废水，2002，33(3)：1～4
[45]Uemoto H, saiki H. Behavior of immobilized nitrosomonas European and paracoccus denitrificans in tubular gel for nitrogen removal in waste water. Pro. Biotechnol, 1996,11(immobilized cells): 695～701