土壤中反硝化细菌的分离与脱氮性能初步研究
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摘要
使用稀释平板法对土壤中微生物进行分离培养,共得到不同形态菌株20株,使用硝酸盐葡萄糖反硝化培养基进行培养,格里斯试剂和二苯胺试剂对菌株进行反硝化显色筛选,并经含有溴百里酚蓝指示剂的反硝化鉴定培养基验证,共得到具有反硝化效果的菌株6株,分别标记为QLLGFXH01-QLLGFXH06。对其进行了形态观察和理化性质研究,监测了在不同碳源条件下的生长曲线,探讨了各个菌株生长的最适碳源,给出了最适碳源下生长曲线的Logistic拟合结果,并对各菌株在葡萄糖为碳源的DM培养基中的反硝化性能进行了研究。结果表明3、4、5三株反硝化细菌其反硝化能力较强,72 h硝酸盐氮去除百分率分别达到了90. 57%、97. 27%和96. 80%。5号在发酵24～48 h和6号在发酵48～72 h出现中间产物亚硝酸盐氮积累,其最高积累量分别为0. 059 mg/L和0. 24 mg/L,其余菌株在硝酸盐氮去除过程中未出现亚硝酸盐氮的积累。总氮去除研究表明1、2、3、6号的72 h总氮去除率均在70%左右,脱氮性能较好。
20 different strains were separated by using dilution plate method,and 6 strains among which were proven to be denitrifying strains after having been cultured with nitrate glucose denitrification medium and verified with Griess reagent,diphenylamine regent and bromothymol blue indicator. The 6 denitrifying strains were then marked as QLLGFXH01-06. The morphology,physicochemical property,growth curve under different carbon sources was then studied. The optimum carbon source and growth curve fitting under optimum carbon were obtained. And the denitrification capability of each strain in DM medium with glucose as carbon source was also studied. The result indicated that strain 3,4 and 5 has strong capability of denitrification,with 72 h nitrate nitrogen removal rate up to 90. 57%,97. 27% and 96. 80% respectively. Nitrite accumulation occurred in the course of fermentation for strain 5 during the 24-48 h and strain 6 during 48-72 h. The maximum accumulation concentrations of nitrite nitrogen were 0. 059 mg/L and 0. 24 mg/L respectively. Strain 1,2,3,6 showed a good denitrification performance with TN removal rate of 70% approximately.
20 different strains were separated by using dilution plate method,and 6 strains among which were proven to be denitrifying strains after having been cultured with nitrate glucose denitrification medium and verified with Griess reagent,diphenylamine regent and bromothymol blue indicator. The 6 denitrifying strains were then marked as QLLGFXH01-06. The morphology,physicochemical property,growth curve under different carbon sources was then studied. The optimum carbon source and growth curve fitting under optimum carbon were obtained. And the denitrification capability of each strain in DM medium with glucose as carbon source was also studied. The result indicated that strain 3,4 and 5 has strong capability of denitrification,with 72 h nitrate nitrogen removal rate up to 90. 57%,97. 27% and 96. 80% respectively. Nitrite accumulation occurred in the course of fermentation for strain 5 during the 24-48 h and strain 6 during 48-72 h. The maximum accumulation concentrations of nitrite nitrogen were 0. 059 mg/L and 0. 24 mg/L respectively. Strain 1,2,3,6 showed a good denitrification performance with TN removal rate of 70% approximately.
引文
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[8]郭龙杰.好氧反硝化菌株Enterobacter cloacae strain HNR的分离筛选及脱氮性能研究[D].重庆:重庆大学,2016.
[9]环境保护部. HJ 636-2012水质总氮的测定碱性过硫酸钾分光光度法[S].北京:中国环境科学出版社,2012.
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[12]刘万鑫,杜荣斌,由翠荣,等.几株异养硝化细菌与好氧反硝化细菌的分离与鉴定[J].烟台大学学报(自然科学与工程版),2017,30(3):249-254.
[13]王有乐,雷兴龙,陈连军,等.土壤中好氧反硝化菌的分离及脱氮特性研究[J].环境工程学报,2011,05(8):1902-1906.
[2]左薇.一株好氧反硝化菌的筛选鉴定及其脱氮特性分析[D].哈尔滨:哈尔滨工业大学,2006.
[3] Takaya N,Catalan-Sakairi M A,Sakaguchi Y,et al.Aerobic denitrifying bacteria that produce low levels of nitrous oxide[J]. Appl Environ Microbiol,2003,69(6):3152-3157.
[4] Zhang H H,Zhou S L. Screening and Cultivation of Oligotrophic Aerobic Denitrifying Bacteria[M]//Zhang H H,Zhou S L. eds. The Handbook of Environmental Chemistry. Cham:Springer International Publishing,2016:451-473. DOI:10. 1007/978-3-319-20391-1_13.
[5] Yang L,Ren Y X,Zhao S Q,et al. Isolation and characterization of three heterotrophic nitrifying-aerobic denitrifying bacteria from a sequencing batch reactor[J].Ann Microbiol,2016,66(2):737-747.
[6]王有乐,陈连军,雷兴龙,等.土壤中好氧反硝化菌分离研究[J].科技信息,2010(1):783-784.
[7]李文君.同步硝化反硝化脱氮效果研究[D].太原:太原理工大学,2006.
[8]郭龙杰.好氧反硝化菌株Enterobacter cloacae strain HNR的分离筛选及脱氮性能研究[D].重庆:重庆大学,2016.
[9]环境保护部. HJ 636-2012水质总氮的测定碱性过硫酸钾分光光度法[S].北京:中国环境科学出版社,2012.
[10]国家环境保护局. GB 7480-87水质硝酸盐氮的测定酚二磺酸分光光度法[S].北京:中国环境科学出版社,1987.
[11]国家环境保护局. GB 7493-87水质亚硝酸盐氮的测定分光光度法[S].北京:中国环境科学出版社,1987.
[12]刘万鑫,杜荣斌,由翠荣,等.几株异养硝化细菌与好氧反硝化细菌的分离与鉴定[J].烟台大学学报(自然科学与工程版),2017,30(3):249-254.
[13]王有乐,雷兴龙,陈连军,等.土壤中好氧反硝化菌的分离及脱氮特性研究[J].环境工程学报,2011,05(8):1902-1906.
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