一株高效好氧反硝化细菌的分离鉴定及脱氮性能研究
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摘要
从广州市西朗珠江水产研究所养殖池中分离新型好氧反硝化细菌,旨为开发新型生物脱氮工艺奠定基础。利用3种不同筛选培养基分离好氧反硝化细菌,测定在单一氮源培养基中发酵的生长曲线和脱氮性能,通过形态和16S rRNA基因序列分析对菌株进行鉴定并构建系统发育树。分离得到一株高性能好氧反硝化菌株BB-17,该菌株在单一氮源培养基中以28℃,180 r/min振荡培养时,其对数期出现在6-24 h,氨氮去除率在18 h达93.92%,硝态氮去除率在48 h达66.11%,亚硝态氮去除率在48h达99.10%。BB-17为革兰氏阴性短杆菌,16S rDNA序列分析结果与Pseudomonas vancouverensis(AJ011507)比对具有98.41%同源性,确定BB-17为假单胞菌属并命名为Pseudomonas sp. BB17。
New types of aerobic denitrifying bacteria were isolated from the culture pond of the Xilang Pearl River Aquaculture Institute in Guangzhou,aiming at laying the foundation for the development of a new biological nitrogen removal process. Three different screening media were used to isolate the aerobic denitrifying bacteria,and the growth curve and denitrification performance of fermentation in a sole nitrogen source medium were determined. Then morphology and 16 S rRNA gene sequence analysis were employed to identify the strains and a phylogenetic tree was constructed. As result,a high-performance aerobic denitrifying strain BB-17 was isolated. When the strain was cultured at 28℃,180 r/min in a sole nitrogen source medium,the logarithmic phase appeared at 6-24 h and the removal rate of ammonia nitrogen was up to 93.92% at 18 h,that of nitrate nitrogen reached 66.11% at 48 h,and that of nitrite nitrogen reached 99.10% at 48 h. BB-17 was a Gramnegative Brevibacterium,the results of 16 S r DNA sequence analysis presented 98.41% homology with Pseudomonas vancouverensis(AJ011507),and BB-17 was identified as Pseudomonas and named P. sp. BB17.
New types of aerobic denitrifying bacteria were isolated from the culture pond of the Xilang Pearl River Aquaculture Institute in Guangzhou,aiming at laying the foundation for the development of a new biological nitrogen removal process. Three different screening media were used to isolate the aerobic denitrifying bacteria,and the growth curve and denitrification performance of fermentation in a sole nitrogen source medium were determined. Then morphology and 16 S rRNA gene sequence analysis were employed to identify the strains and a phylogenetic tree was constructed. As result,a high-performance aerobic denitrifying strain BB-17 was isolated. When the strain was cultured at 28℃,180 r/min in a sole nitrogen source medium,the logarithmic phase appeared at 6-24 h and the removal rate of ammonia nitrogen was up to 93.92% at 18 h,that of nitrate nitrogen reached 66.11% at 48 h,and that of nitrite nitrogen reached 99.10% at 48 h. BB-17 was a Gramnegative Brevibacterium,the results of 16 S r DNA sequence analysis presented 98.41% homology with Pseudomonas vancouverensis(AJ011507),and BB-17 was identified as Pseudomonas and named P. sp. BB17.
引文
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[10]颜薇芝. 1株异养硝化好氧反硝化不动杆菌的分离及脱氮性能[J].环境工程学报, 2017, 11(7):4419-4428.
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[14]战海.一株好氧反硝化细菌的分离鉴定及反硝化能力初测[J].天津农学院学报, 2017, 24(2):44-48.
[15]丁梦娇.植烟土壤中微生物特性及氨化、亚硝化菌分离鉴定与活性研究[J].中国生态农业学报, 2017, 25(10):1444-1455.
[16]牟东阳,靳鹏飞,彭永臻,等. 1株异养硝化-好氧反硝化细菌DK1的分离鉴定及其脱氮特性[J].环境科学, 2017, 38(11):4763-4773.
[17]沈辉,万夕和,蒋葛.两株海洋异养好氧硝化-反硝化细菌的筛选、鉴定及能力测试[J].应用与环境生物学报, 2017, 23(1):157-163.
[18]菜钰颖,商平,魏丽娜,等.氨氮降解微生物菌株的分离筛选及去除效果初步研究[J].净水技术, 2008, 27(3):44-47.
[19]刘思雅.高氨氮条件碳源对地衣芽孢杆菌(Dy)降氮的影响[J].江西农业大学学报, 2017, 39(5):1002-1009.
[20]黄婧.畜禽养殖污水中高效氨氮降解菌的筛选、鉴定及生长条件研究[J].环境工程学报, 2011, 8(5):1779-1784.
[21]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, 5(8):1902-1906.
[2] Paerl HW, Gardner WS, Macarthy MJ, et al. Algal blooms:noteworth nitrogen[J]. Science, 2014, 346(6206):175.
[3] Vitousek PM, Aber JD, Howarth RW, et al. Human alteration of the global nitrogen cycle:sources and consequences[J]. Ecological Applications, 1997, 7(3):737-750.
[4]彭志兰,柳敏海,郭海波,等.一株高效好氧反硝化菌的分离与鉴定[J].上海海洋大学学报, 2015, 24(4):632-639.
[5]李卫芬. 1株好氧反硝化菌的分离鉴定及反硝化特性研究[J].环境科学, 2011, 32(8):2403-2408.
[6]孙锦宜.含氮废水处理技术与应用[M].北京:化学工业出版社,2003.
[7] Frette L, Gejlsbjerg B, Westermann P. Aerobic denitrifiers isolated from an alternating active sludge system[J]. FEMS Microbiology Ecology, 1997, 24:363-370.
[8] Robertson LA, Kuenen JG. Aerobic denitrification:A controversy revived[J]. Archives of Microbiology, 1984, 139(4):351-354.
[9] Kundu P, Pramanik A, Dasgupta A, et al. Simultaneous heterotrophic nitrification and aerobic denitrification by Chryseobacterium sp. R31 isolated from abattoir wastewater[J]. BioMed Research Internationa, 2014. doi:10. 1155/2014/436056.
[10]颜薇芝. 1株异养硝化好氧反硝化不动杆菌的分离及脱氮性能[J].环境工程学报, 2017, 11(7):4419-4428.
[11] Srinandan CS, Shah M, Patal B, et al. Assessment of denitrifying bacterial composition in activated sludge[J]. Bioresource Technology, 2011, 102(20):9481-9489.
[12]成钰,李秋芬,费聿涛,等.海水异养硝化-好氧反硝化芽孢杆菌SLWX2的筛选及脱氮特性[J].环境科学, 2016(37):2681-2688.
[13]王田野,魏荷芬,胡子全,等.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, 2017(3):946-953.
[14]战海.一株好氧反硝化细菌的分离鉴定及反硝化能力初测[J].天津农学院学报, 2017, 24(2):44-48.
[15]丁梦娇.植烟土壤中微生物特性及氨化、亚硝化菌分离鉴定与活性研究[J].中国生态农业学报, 2017, 25(10):1444-1455.
[16]牟东阳,靳鹏飞,彭永臻,等. 1株异养硝化-好氧反硝化细菌DK1的分离鉴定及其脱氮特性[J].环境科学, 2017, 38(11):4763-4773.
[17]沈辉,万夕和,蒋葛.两株海洋异养好氧硝化-反硝化细菌的筛选、鉴定及能力测试[J].应用与环境生物学报, 2017, 23(1):157-163.
[18]菜钰颖,商平,魏丽娜,等.氨氮降解微生物菌株的分离筛选及去除效果初步研究[J].净水技术, 2008, 27(3):44-47.
[19]刘思雅.高氨氮条件碳源对地衣芽孢杆菌(Dy)降氮的影响[J].江西农业大学学报, 2017, 39(5):1002-1009.
[20]黄婧.畜禽养殖污水中高效氨氮降解菌的筛选、鉴定及生长条件研究[J].环境工程学报, 2011, 8(5):1779-1784.
[21]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, 5(8):1902-1906.