高氨氮废水处理系统中功能微生物的检测

Detection and Analysis of Functional Microbes in Strong Ammonia Wastewater Treatment System

利用16S rDNA分子检测技术对高氨氮废水处理系统中的氨氧化细菌、亚硝酸盐氧化细菌等难分离培养微生物进行了分析检测.从实验室处理高氨氮废水的生物流化床硝化系统生物膜中提取细菌总DNA,以其为模板,利用氨氧化细菌(AOB)和亚硝酸盐氧化细菌(NOB)的16S rDNA特异性引物进行多聚酶链式反应(PCR)扩增,获得AOB和NOB的特异片段.将特异片段与pGEM-T载体连接并转化到E.coli DH5α中获得重组子,对阳性重组子进行酶切分型,AOB可分为8种不同类型.选取AOB的8种类型代表以及随即选取8个NOB克隆进行测序.经GenBank搜索以及同源性分析,发现所获得的8个AOB类型来自不同的菌株,处理系统中以Nitrosomonas sp.和Nitrosococcus sp.为氨氧化细菌的优势菌属;8株NOB重组子中有7株来自不同的菌株,系统中Nitrobacter sp.和Afipia sp.为亚硝酸盐氧化细菌的优势菌属.

Total DNA was extracted from the nitrifying biofilm in strong ammonia wastewater treatment system, and two pairs of polymerase chain reaction （PCR） primers based on the 16S rDNA sequences of ammonia-oxidizing bacteria （AOB） and nitriteoxidizing bacteria （NOB） were used for PCR amplification. The special fragments of AOB and NOB were cloned into E. coli DH5α, and after digested by different restriction endonucleases, 8 strains of AOB and 8 strains of NOB were sequenced. The 8 AOB clones were found coming from different strains by comparing with the DNA sequences in GenBank （ NCBI）, and Nitrosomonas sp. and Nitrosococcus sp. were the dominant species for ammonia oxidation in the system. The 8 NOB clones came from 7 different strains, and Nitrobacter sp. and Afipia sp. were the dominant species for nitrite oxidation. Fig 6, Tab 1, Ref 14