一株好氧反硝化细菌的分离与鉴定

doi:10.16431/j.cnki.1671-7236.2015.07.037

›› 2015, Vol. 42 ›› Issue (7): 1877-1882.doi: 10.16431/j.cnki.1671-7236.2015.07.037

一株好氧反硝化细菌的分离与鉴定

郑建龙¹, 时书宁¹, 雍燕², 方炳虎¹

1. 华南农业大学兽医学院, 广州 510642;
2. 广东大华农动物保健品股份有限公司, 云浮 527400

收稿日期:2015-01-20 出版日期:2015-07-20 发布日期:2015-07-23
通讯作者: 方炳虎 E-mail:fangbh@scau.edu.cn
作者简介:郑建龙(1989-),男,福建漳平人,硕士生,研究方向:兽医药理学与毒理学,E-mail:545057302@qq.com
基金资助:
国家自然科学基金(31172359)

Isolation and Identification of an Aerobic Denitrifying Bacterium

ZHENG Jian-long¹, SHI Shu-ning¹, YONG Yan², FANG Bing-hu¹

1. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;
2. Guangdong Dahuanong Animal Health Products Co., Ltd., Yunfu 527400, China

Received:2015-01-20 Online:2015-07-20 Published:2015-07-23

摘要/Abstract

摘要： 本试验旨在从环境样品筛选出一株具有去除氨氮和亚硝酸盐氮的好氧反硝化细菌。通过对采集水样进行富集驯化,利用溴百里酚蓝(BTB)选择性培养基进行初筛,并以去除氨氮、亚硝酸盐氮能力为指标进行复筛,得到一株好氧反硝化细菌DG-3。在好氧条件下测定了菌株DG-3的反硝化能力,并对其进行了形态学观察、生理生化试验和16S rDNA基因序列的分析。菌株DG-3在好氧反硝化培养过程中对硝酸盐氮(138.63 mg/L)的去除率达75.57%,在整个培养过程中,仅出现了少量的亚硝酸盐氮累积,其最大累积值仅为2.06 mg/L。反硝化培养基培养及扫描电镜观察结果显示,菌株DG-3为杆状,无鞭毛,大小为(1.7~2.8)μm×(0.6~0.7)μm,菌落呈淡黄色。结合形态学、生理生化特性及16S rDNA同源性比对分析,初步判断菌株DG-3为门多萨假单胞菌(Pseudomonas mendocina)。菌株DG-3具有好氧反硝化能力,且能去除氨氮和亚硝酸盐氮,在水产养殖含氮废水的处理中具广阔的应用前景。

关键词: 好氧反硝化; 硝酸盐; 氨氮; 鉴定

Abstract: The present study was aimed to isolate an aerobic denitrifying bacterium from environmental samples.The collecting water samples were enriched and domesticated,the bromothymol blue (BTB) selective medium were used for screening.Based on the aerobic denitrification activity,ammonium and nitrite nitrogen removing ability,the strain DG-3 was isolated from the sample.The strain was identified according to its aerobic denitrification ability,morphological,physiological and biochemical properties and phylogenetic analysis of its 16S rDNA sequence.The results showed that the reduction efficiency of strain DG-3 was up to 75.57% under aerobic conditions with initial NO₃^--N concentration of 138.63 mg/L,and the accumulated nitrite appeared during the process of denitrification,the max concentration was 2.06 mg/L.The morphology of strain DG-3 was with an average size of (1.7 to 2.8)μm×(0.6 to 0.7)μm and a pale yellow surface.DG-3 was identified as Pseudomonas mendocina by morphological and biochemical characteristics and molecular properties.It could be concluded that DG-3 had an aerobic denitrification ability with removing ammonium and nitrite,which showed a great potential in nitrogen wastewater treatment.

Key words: aerobic denitrification; nitrate; ammonium; identification

中图分类号:

X172

郑建龙, 时书宁, 雍燕, 方炳虎. 一株好氧反硝化细菌的分离与鉴定[J]. , 2015, 42(7): 1877-1882.

ZHENG Jian-long, SHI Shu-ning, YONG Yan, FANG Bing-hu. Isolation and Identification of an Aerobic Denitrifying Bacterium[J]. , 2015, 42(7): 1877-1882.

参考文献

[1] Rodrigues R V,Schwarz M H,Delbos B C,et al.Acute toxicity and sublethal effects of ammonia and nitrite for juvenile cobia Rachycentron canadum[J].Aquaculture,2007,271(1):553-557.
[2] Lewis Jr W M,Morris D P.Toxicity of nitrite to fish:A review[J].Transactions of the American Fisheries Society,1986,115(2):183-195.
[3] Arillo A,Margiocco C,Melodia F,et al.Ammonia toxicity mechanism in fish:Studies on rainbow trout (Salmo gairdneri Rich)[J].Ecotoxicology and Environmental Safety,1981,5(3):316-328.
[4] Shi Z,Zhang Y,Zhou J,et al.Biological removal of nitrate and ammonium under aerobic atmosphere by Paracoccus versutus LYM[J].Bioresource Technology,2013,148:144-148.
[5] Khardenavis A A,Kapley A,Purohit H J.Simultaneous nitrification and denitrification by diverse Diaphorobacter sp.[J].Applied Microbiology and Biotechnology,2007,77(2):403-409.
[6] 郭端强,刘海龙,万亚涛,等.一株好氧反硝化细菌的分离鉴定及反硝化特性研究[J].生物技术通报,2012,10:205-209.
[7] Chen Q,Ni J.Ammonium removal by Agrobacterium sp.LAD9 capable of heterotrophic nitrification-aerobic denitrification[J].Journal of Bioscience and Bioengineering,2012,113(5):619-623.
[8] Zhang J,Wu P,Hao B,et al.Heterotrophic nitrification and aerobic denitrification by the bacterium Pseudomonas stutzeri YZN-001[J].Bioresource Technology,2011,102(21):9866-9869.
[9] Song Z,An J,Fu G,et al.Isolation and characterization of an aerobic denitrifying Bacillus sp.YX-6 from shrimp culture ponds[J].Aquaculture,2011,319(1):188-193.
[10] Robertson L A,van Niel E W,Torremans R A,et al.Simultaneous nitrification and denitrification in aerobic chemostat cultures of Thiosphaera pantotropha[J].Appl Environ Microbiol,1988,54(11):2812-2818.
[11] 杨小龙,李文明,陈燕,等.一株好氧反硝化菌的分离鉴定及其除氮特性[J].微生物学报,2011,8:1062-1070.
[12] 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.
[13] 郑巧东,钟丽娜,姚善泾.硝化菌与反硝化菌混合培养生物脱氮的研究[J].化学工程,2010,3:64-67.
[14] 东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[15] Buchanan R E.伯杰细菌鉴定手册[M].第8版.北京:科学出版社,1984.
[16] 项慕飞.好氧反硝化菌的分离筛选和鉴别研究[D].北京:北京工商大学,2007.
[17] 复盛,国家环境保护总局,水和废水监测分析方法编委会.水和废水监测分析方法[M].北京:中国环境科学出版社,2002.
[18] Bernat K,Wojnowska-Baryła I.Carbon source in aerobic denitrification[J].Biochemical Engineering Journal,2007,36(2):116-122.
[19] 王有乐,雷兴龙,王亚变,等.好氧反硝化菌筛选方法的研究进展[J].安徽农业科学,2010,38(10):5294-5296.
[20] 张小玲,张卫东,张玲,等.好氧反硝化菌的选育及其初步应用[J].微生物学通报,2008,35(10):1556-1561.
[21] 石小彤,李彦芹,邢国伟,等.一株耐盐好氧反硝化细菌的分离筛选及鉴定[J].生物技术通报,2013,3:175-180.
[22] 郑佳佳,沈涛,傅罗琴,等.一株硝化反硝化菌的筛选鉴定及反硝化特性研究[J].水生生物学报,2012,36(1):161-167.
[23] 褚淑祎,江惠霞,肖继波,等.高效反硝化菌 aHD7 的筛分、脱氮特性及厌氧氨氧化性[J].应用生态学报,2012,23(11):3096.
[24] 李卫芬,张小平,宋文辉,等.芽孢杆菌对草鱼养殖水质调控作用的研究[J].渔业现代化,2011,4:22-26.
[25] 陶蕾,梁剑平,郭凯,等.复合活菌制剂降解虾塘中亚硝酸盐的研究与应用[J].中国畜牧兽医,2011,38(8):182-184.

一株好氧反硝化细菌的分离与鉴定

Isolation and Identification of an Aerobic Denitrifying Bacterium

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