Jiang He-Long, Tay Joo-Hwa, Maszenan Abdul Majid, Tay Stephen Tiong-Lee
Environmental Engineering Research Centre, School of Civil and Environmental Engineering, Nanyang Technological University, Singapore.
Appl Environ Microbiol. 2004 Nov;70(11):6767-75. doi: 10.1128/AEM.70.11.6767-6775.2004.
Aerobic granules are self-immobilized aggregates of microorganisms and represent a relatively new form of cell immobilization developed for biological wastewater treatment. In this study, both culture-based and culture-independent techniques were used to investigate the bacterial diversity and function in aerobic phenol- degrading granules cultivated in a sequencing batch reactor. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes demonstrated a major shift in the microbial community as the seed sludge developed into granules. Culture isolation and DGGE assays confirmed the dominance of beta-Proteobacteria and high-G+C gram-positive bacteria in the phenol-degrading aerobic granules. Of the 10 phenol-degrading bacterial strains isolated from the granules, strains PG-01, PG-02, and PG-08 possessed 16S rRNA gene sequences that matched the partial sequences of dominant bands in the DGGE fingerprint belonging to the aerobic granules. The numerical dominance of strain PG-01 was confirmed by isolation, DGGE, and in situ hybridization with a strain-specific probe, and key physiological traits possessed by PG-01 that allowed it to outcompete and dominate other microorganisms within the granules were then identified. This strain could be regarded as a functionally dominant strain and may have contributed significantly to phenol degradation in the granules. On the other hand, strain PG-08 had low specific growth rate and low phenol degradation ability but showed a high propensity to autoaggregate. By analyzing the roles played by these two isolates within the aerobic granules, a functional model of the microbial community within the aerobic granules was proposed. This model has important implications for rationalizing the engineering of ecological systems.
好氧颗粒是微生物的自固定聚集体,代表了一种为生物废水处理而开发的相对新型的细胞固定形式。在本研究中,基于培养和不依赖培养的技术都被用于研究在序批式反应器中培养的好氧苯酚降解颗粒中的细菌多样性和功能。对PCR扩增的16S rRNA基因进行变性梯度凝胶电泳(DGGE)分析表明,随着种子污泥发育成颗粒,微生物群落发生了重大转变。培养分离和DGGE分析证实了β-变形菌和高G+C革兰氏阳性菌在苯酚降解好氧颗粒中的优势地位。从颗粒中分离出的10株苯酚降解细菌菌株中,PG-01、PG-02和PG-08菌株的16S rRNA基因序列与属于好氧颗粒的DGGE指纹图谱中优势条带的部分序列相匹配。通过分离、DGGE和用菌株特异性探针进行原位杂交,证实了PG-01菌株在数量上的优势,然后确定了PG-01菌株所具有的使其能够在颗粒中胜过并主导其他微生物的关键生理特性。该菌株可被视为功能优势菌株,可能对颗粒中的苯酚降解有显著贡献。另一方面,PG-08菌株的比生长速率低且苯酚降解能力低,但表现出较高的自聚集倾向。通过分析这两种分离物在好氧颗粒中的作用,提出了好氧颗粒内微生物群落的功能模型。该模型对于合理设计生态系统工程具有重要意义。