草坪土壤中好氧反硝化菌的分离筛选及其脱氮特性的研究
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摘要
根据历年《中国环境状况公报》显示,我国水体中氮素污染日益严重,对自然环境和人体健康产生极大危害,氮素在水体中主要以有机氮和无机氮两种形式存在。废水脱氮方法很多,其中生物脱氮是目前最经济有效的方法之一。因传统生物脱氮中各脱氮步骤的不同细菌菌群对营养物质及溶解氧的要求不同,工艺复杂,极大限制了生物脱氮在实际工程中的应用。近年国内外不少研究充分证明了好氧反硝化菌的存在,突破了传统生物脱氮的理论,为生物脱氮技术及其应用提供了崭新的思路与发展。与传统的缺氧反硝化脱氮相比,利用好氧反硝化菌发展生物脱氮技术具有独特优势,已成为废水脱氮研究的热点。
本实验采用极限稀释、滴加格里斯试剂及二苯胺试剂鉴别、平板划线相结合的方法,从校园草坪土壤中分离筛得一株能以硝酸钾为氮源进行好氧反硝化作用的好氧反硝化菌LD3。对菌株LD3进行种属鉴定,并研究其反硝化能力及各种环境因素的影响情况,包括碳源、氮源、溶解氧、C/N、pH值和温度等。研究结果表明,菌株LD3初步鉴定为脂肪杆菌属,其硝酸盐氮去除率高达97.67%,最佳脱氮时间为5d,最适合菌液剂量比例为5%；除过高浓度的亚硝酸盐对菌株LD3生长繁殖有一定的抑制作用外,菌株LD3具有较广泛的碳源谱和氮源谱；菌株LD3对生长环境的溶解氧要求不高,浓度达到1.8mg/L即可,其好氧反硝化作用最佳的初始C/N为8,最佳的初始pH值为7.5,最适宜的温度为30℃左右。正交实验确定菌株LD3好氧反硝化作用最优的环境因素条件为：转速为160r/min,初始C/N为10,初始pH值为7.5,温度为30℃。菌株LD3的分离筛选及其脱氮特性的实验研究对实现好氧反硝化菌脱氮技术利用提供了理论参考。
According to the Report on the State of Environment in China over the years, increasingly severe nitrogen pollution of water bodies does great harm to the natural environment and human health, in which nitrogen was present in the organic and inorganic forms.One of economic and effective approachs among numerous nitrogen removal processes was biodenitrification, yet which was confined in practice due to the complex process, the various nutriment and dissolved oxygen (DO) taken by corresponding bacterias in the diverse traditional nitrogen removal steps. The existence of aerobic denitrifier, proved by the domestic and international research in recent decades, made a breakthrough at the tradition denitrification theory, provided a new perspective to view the biodenitrification technology and application. Compared with the tradition anoxic denitrification, aerobic denitrification had became a hotspot of nitrogen removal from wastewater owing to its unique superiority.
The aerobic denitrifier strain LD3with potassium nitrate as nitrogen source was isolated and screened from the campus lawn soil via limiting concentration, adding Chris reagents and diphenylamine and combined with plate streaking, and then expertized species identification, investigated denitrification ability and the effect of environment factors including carbon source, nitrogen source, DO, carbon nitrogen ratio (C/N), pH,temperature, etc. It was concluded that strain LD3was preliminarily identified to be pimelobacter, The removal rate of nitratenitrogen reached97.67%, the best denitrification time was5days, the optimum bacterium fluid does ratio was5%.Strain LD3lacked advantage of rich carbon and nitrogen sources except that high concentration nitrite was a disincentive. Strain LD3wasn't strict with DO, which could meet the demands at1.8mg/L. The optimum initial C/N, pH and temperature was8,7.5and30℃respectively for aerobic denitrification. the best environmental factors determined by orthogonal test was as follows:rotate speed, initial C/N, initial pH, temperature was160r/min,10,7.5and30℃respectively. Experiemntal investigation on identification and denitrification characteration of strain LD3provided the theoretical reference for achieving nitrogen removal by aerobic denitrifier.
本实验采用极限稀释、滴加格里斯试剂及二苯胺试剂鉴别、平板划线相结合的方法,从校园草坪土壤中分离筛得一株能以硝酸钾为氮源进行好氧反硝化作用的好氧反硝化菌LD3。对菌株LD3进行种属鉴定,并研究其反硝化能力及各种环境因素的影响情况,包括碳源、氮源、溶解氧、C/N、pH值和温度等。研究结果表明,菌株LD3初步鉴定为脂肪杆菌属,其硝酸盐氮去除率高达97.67%,最佳脱氮时间为5d,最适合菌液剂量比例为5%；除过高浓度的亚硝酸盐对菌株LD3生长繁殖有一定的抑制作用外,菌株LD3具有较广泛的碳源谱和氮源谱；菌株LD3对生长环境的溶解氧要求不高,浓度达到1.8mg/L即可,其好氧反硝化作用最佳的初始C/N为8,最佳的初始pH值为7.5,最适宜的温度为30℃左右。正交实验确定菌株LD3好氧反硝化作用最优的环境因素条件为：转速为160r/min,初始C/N为10,初始pH值为7.5,温度为30℃。菌株LD3的分离筛选及其脱氮特性的实验研究对实现好氧反硝化菌脱氮技术利用提供了理论参考。
According to the Report on the State of Environment in China over the years, increasingly severe nitrogen pollution of water bodies does great harm to the natural environment and human health, in which nitrogen was present in the organic and inorganic forms.One of economic and effective approachs among numerous nitrogen removal processes was biodenitrification, yet which was confined in practice due to the complex process, the various nutriment and dissolved oxygen (DO) taken by corresponding bacterias in the diverse traditional nitrogen removal steps. The existence of aerobic denitrifier, proved by the domestic and international research in recent decades, made a breakthrough at the tradition denitrification theory, provided a new perspective to view the biodenitrification technology and application. Compared with the tradition anoxic denitrification, aerobic denitrification had became a hotspot of nitrogen removal from wastewater owing to its unique superiority.
The aerobic denitrifier strain LD3with potassium nitrate as nitrogen source was isolated and screened from the campus lawn soil via limiting concentration, adding Chris reagents and diphenylamine and combined with plate streaking, and then expertized species identification, investigated denitrification ability and the effect of environment factors including carbon source, nitrogen source, DO, carbon nitrogen ratio (C/N), pH,temperature, etc. It was concluded that strain LD3was preliminarily identified to be pimelobacter, The removal rate of nitratenitrogen reached97.67%, the best denitrification time was5days, the optimum bacterium fluid does ratio was5%.Strain LD3lacked advantage of rich carbon and nitrogen sources except that high concentration nitrite was a disincentive. Strain LD3wasn't strict with DO, which could meet the demands at1.8mg/L. The optimum initial C/N, pH and temperature was8,7.5and30℃respectively for aerobic denitrification. the best environmental factors determined by orthogonal test was as follows:rotate speed, initial C/N, initial pH, temperature was160r/min,10,7.5and30℃respectively. Experiemntal investigation on identification and denitrification characteration of strain LD3provided the theoretical reference for achieving nitrogen removal by aerobic denitrifier.
引文
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