两个CANON污水处理系统中氨氧化古菌的丰度和多样性研究

Abundance and Community Composition of Ammonia-Oxidizing Archaea in Two Completely Autotrophic Nitrogen Removal over Nitrite Systems

近期,氨氧化古菌(ammonia-oxidizing archaea,AOA)在各类环境中的发现,打破了人们原来认为氨氧化主要是由氨氧化细菌(ammonia-oxidizing bacteria,AOB)完成的观点.但是在全程自养脱氮(completely autotrophic nitrogen removal over nitrite,CANON)污水处理系统中关于AOA的研究却鲜有提及.利用PCR、克隆、实时荧光定量PCR(quantitative real time PCR,q PCR)等分子生物学技术,对1个小试(lab-scale,L)和1个中试(pilot-scale,P)CANON系统中的生物膜和活性污泥絮体两种形态的污泥进行AOA数量和种属特征的研究.结果表明:1系统L和P中,AOA的amo A基因数量平均值(以dry sludge计)分别为2.42×106copies·g-1和6.51×106copies·g-1;2 AOA的amo A基因丰度随污泥形态不同数量相差很大:L系统中,生物膜AOA amo A丰度约为活性污泥絮体的11.1~15.1倍;P系统中,污泥絮体AOA amo A数量是生物膜中的2.8倍;3多样性方面:P系统的AOA多样性较低,仅出现一个OTU,该OTU属于Nitrosopumilus subcluster 5.2类群;L系统AOA多样性较高,共有8个OTU出现,分别属于Nitrososphaera subcluster 9、subcluster 8.1、subcluster 4.1、subcluster 1.1和Nitrosopumilus subcluster 5.2这5个类群.总之,在同一个CANON系统中,污泥形态不同,AOA的丰度和群落结构相差较大;AOA可能发挥着氨氧化的作用.

Ammonia oxidation is the first and rate-limiting step of nitrification,which was thought to be only performed by ammoniaoxidizing bacteria( AOB). In recent years,ammonia-oxidizing archaea( AOA) was also confirmed to take part in ammonia oxidation.The diversity and abundance of AOA have been investigated in various environments,however,little is known regarding the AOA in the completely autotrophic nitrogen removal over nitrite( CANON) wastewater treatment process. In this study,the abundance and diversity of AOA were investigated in the biofilm and flocculent activated sludge collected in a lab-scale( L) CANON system and a pilot-scale( P) CANON systems,respectively. The quantitative real time PCR( q PCR) was applied to investigate the abundance of AOA and the diversity of AOA was determined by polymerase chain reaction( PCR),cloning and sequencing. The q PCR results showed that the average abundance of AOA amo A gene of L and P was 2. 42 × 106copies·g-1dry sludge and 6. 51 × 106copies·g-1dry sludge,respectively. The abundance of AOA in biofilm was 10. 1-14. 1 times higher than that in flocculent activated sludge. For P system,the abundance of AOA in flocculent activated sludge was 1. 8 times higher than that in biofilm. The results indicated that the abundance of AOA might be affected by different sludge morphology. The diversity of AOA in P system was extremely limited,only one OTU was observed,which was classified into Nitrosopumilus subcluster 5. 2. The diversity of AOA in L system was higher,eight OTUs were observed,which were classified into five genera: Nitrososphaera subcluster 9,subcluster 8. 1,subcluster 4. 1,subcluster 1. 1 and Nitrosopumilus subcluster 5. 2. The diversity and abundance of AOA were different in CANON systems with different sludge morphology. AOA may play an important role in ammonia oxidation in CANON system.