几株异养硝化细菌与好氧反硝化细菌的分离与鉴定
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摘要
从海水养殖场的生物膜,逛荡河口淤泥,烟大三元湖底泥中分离筛选出8株异养硝化细菌和8株好氧反硝化细菌.分别对16株细菌的异养硝化性能和好氧反硝化性能进行测定,结果表明,异养硝化细菌Y-3、Y-6、Y-7的72 h NH4+-N去除率分别为95.65%、96.60%、96.09%,好氧反硝化细菌F-3、F-5、F-7的72 h NO3--N去除率分别为76.20%、75.87%、76.00%.选择性能较高的3株异养硝化细菌和3株好氧反硝化细菌进行传统生理生化鉴定,并对各菌株的16S r DNA序列进行测定及分析,结果表明,Y-3和F-3为除烃海杆菌(Marinobacter hydrocarbonoclasticus),Y-6为溶藻孤菌(Vibrio alginolyticus),Y-7和F-7为恶臭假单胞菌(Pseudomonas putida),F-5为海洋单胞菌属(Oceanimonas sp.).
Eight strains of heterotrophic nitrifying bacteria and eight strains of aerobic denitrification bacteria are isolated from the sediment and biofilm samples collected from the recirculating aquaculture system,Sanyuan Lake and Guangdang River in Yantai. The heterotrophic nitrification and aerobic denitrification performance of the 16 strains are tested. It is shown that the NH+4-N removal rates in 72 h for heterotrophic nitrifying bacteria strains Y-3,Y-6 and Y-7 are 95. 65%,96. 60% and 96. 09% respectively,and the NO-3-N removal rates in 72 h for aerobic denitrifying bacteria strains F-3,F-5 and F-7 are 76. 20%,75. 87% and 76. 00% respectively. Six strains,Y-3,Y-6,Y-7,F-3,F-5 and F-7,with higher capability are identified. The 16 S r DNA sequences of the six selected strains are determined and aligned with those in the Gen Bank database. Strains Y-3 and F-3 are identified as Marinobacter hydrocarbonoclasticus,strain Y-6 is identified as Vibrio alginolyticus,strains Y-7 and F-7 are identified as Pseudomonas putida,and strain F-5 is identified as Oceanimonas sp.
Eight strains of heterotrophic nitrifying bacteria and eight strains of aerobic denitrification bacteria are isolated from the sediment and biofilm samples collected from the recirculating aquaculture system,Sanyuan Lake and Guangdang River in Yantai. The heterotrophic nitrification and aerobic denitrification performance of the 16 strains are tested. It is shown that the NH+4-N removal rates in 72 h for heterotrophic nitrifying bacteria strains Y-3,Y-6 and Y-7 are 95. 65%,96. 60% and 96. 09% respectively,and the NO-3-N removal rates in 72 h for aerobic denitrifying bacteria strains F-3,F-5 and F-7 are 76. 20%,75. 87% and 76. 00% respectively. Six strains,Y-3,Y-6,Y-7,F-3,F-5 and F-7,with higher capability are identified. The 16 S r DNA sequences of the six selected strains are determined and aligned with those in the Gen Bank database. Strains Y-3 and F-3 are identified as Marinobacter hydrocarbonoclasticus,strain Y-6 is identified as Vibrio alginolyticus,strains Y-7 and F-7 are identified as Pseudomonas putida,and strain F-5 is identified as Oceanimonas sp.
引文
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[19]陈垚,黄鹏程,杨威,等.好氧颗粒污泥处理高盐榨菜废水除污特性研究[J].工业水处理,2015,35(11):29-32.
[20]巫亮,李荣鹏,訾小利,等.厌氧及好氧条件下盐度对海杆菌NY-9生长及反硝化能力的影响[J].湖北农业科学,2014,53(23):5705-5708
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[22]黄菲菲,张晋华.异养硝化-好氧反硝化菌的筛选与脱氮性能研究[D].南京:南京理工大学,2013.
[2]王欢,汪苹,刘晶晶.好氧反硝化菌的异养硝化性能研究[J].环境科学与技术,2008,31(11):45-47.
[3]尹丽,肖小雨,梁亮亮,等.一株异养硝化细菌的分离鉴定及其脱氮效果[J].环境科学与技术,2014,37(12):25-30.
[4]苟莎,黄钧.异养硝化细菌脱氮特性及研究进展[J].微生物学通报,2009,36(2):255-260.
[5]王洁,蓝江林,刘波.一株异养硝化细菌的分离鉴定和脱氮特性研究[J].农业环境科学学报,2013,32(4):805-810.
[6]PAI S L,CHONG N M,CHEN C H.Potentiala Applications of aerobic denitrifiying bacteria as bio-agents in wastewater treatment[J].Bioresource Technology,1999,68(2):179-185.
[7]潘丹,黄巧云,陈雯莉.两株异养硝化细菌的分离鉴定及其脱氮特性[J].微生物学报,2011,51(10):1382-1389.
[8]BRIERLEY E D R,WOOD M.Heterotrophic nitrification in an acid forest soil:isolation and characterisation of a nitrifying bacterium[J].Soil Biol Biochem,2001,33:1403-1409.
[9]VELUSAMY K,KRISHNANI K K.Heterotrophic nitrifying and oxygen tolerant denitrifying bacteria from greenwater system of coastal aquaculture[J].Applied Biochemistry and Biotechnology,2013,169(6):1978-1992.
[10]PATUREAU D,ZUMSTEIN E,DELGENES J P.Aerobic denitrification isolation from diverse natural and managed Ecosystems[J].Microbiol Ecol,2000,39:145-152.
[11]闫志英,廖银章,李旭东,等.新型废水生物脱氮的微生物学研究进展[J].应用与环境生物学报,2006,12(2):292
[12]陈赵芳,尹立红,浦跃朴,等.一株异养硝化菌的筛选及其脱氮条件[J].东南大学学报(自然科学版),2007,37(3):486-490.
[13]国家环境保护总局.水和废水监测分析方法[M].北京:中国环境科学出版社,2000:253-257,279-281.
[14]高喜燕,刘鹰,郑海燕,等.一株海洋好氧反硝化细菌的鉴定及其好氧反硝化特性[J].微生物学报,2010,50(9):1164-1171.
[15]胡甜,江林峰,陈青云,等.高效异养硝化细菌的分离筛选及多样性分析[J].湖北农业科学,2015,54(5):1181-1185.
[16]许尚营,张华,李娜,等.一株好氧反硝化细菌的鉴定及反硝化特性研究[J].环境科学与技术,2010,33(2):10-13.
[17]孙朦朦,陈成勋.两株海水异养硝化-好氧反硝化菌的分离、鉴定、培养、硝化性能及脱氮性能研究[D].天津:天津农学院,2014.
[18]单洪伟,高磊,马甡,等.不同C∶N条件下一株溶藻弧菌对不同种类氮源的吸收特性[J].中国水产科学,2013,20(5):1066-1075.
[19]陈垚,黄鹏程,杨威,等.好氧颗粒污泥处理高盐榨菜废水除污特性研究[J].工业水处理,2015,35(11):29-32.
[20]巫亮,李荣鹏,訾小利,等.厌氧及好氧条件下盐度对海杆菌NY-9生长及反硝化能力的影响[J].湖北农业科学,2014,53(23):5705-5708
[21]NAKANO M,INAGAKI T,OKUNISHI S,et al.Effect of salinity on denitrification under limited single carbon source by Marinobacter sp.isolated from marine sediment[J].Basic Microbiol,2010,50:285-289.
[22]黄菲菲,张晋华.异养硝化-好氧反硝化菌的筛选与脱氮性能研究[D].南京:南京理工大学,2013.