电极活性菌分离过程中微生物群落结构动态特征解析

为探讨培养过程对微生物的选择性作用及电极活性菌在生物膜中的功能地位,采用亨氏厌氧滚管稀释培养技术,以氧化铁为指征,自微生物燃料电池(microbial fuel cell,MFC)阳极生物膜中分离电极活性微生物,并对纯菌分离过程中微生物群落结构动态特征进行跟踪监测.结果表明,以乳酸、乙酸和甾体药物生产废水为底物的MFC最高电压分别可达0.57、0.60和0.40 V.种泥以及乳酸、乙酸为底物的阳极膜分离出的优势菌种均为变形杆菌,而甾体废水为底物的阳极膜分离出的优势菌包含变形杆菌、厚壁菌和拟杆菌,种类丰富.经过富集培养最终分离获得2株电极活性菌,16S rDNA测序鉴定为路德维希肠杆菌(Enterobacter ludwigii)和弗氏柠檬酸杆菌(Citrobacter freundii).以乳酸为底物,两株纯菌可分别获得0.10V和0.17 V的最大电压.该研究表明,应用氧化铁为指征的厌氧梯度稀释培养技术可分离到电极活性微生物,但由于分离过程的选择作用,微生物群落结构动态变化特征显著.

Characteristics of Microbial Community Structure During Isolation of Electrical Active Bacteria

To investigate the effect of selective culturing on microorganisms and functional role of electrical active bacteria in biofilm, some exoelectrogens were isolated from microbial fuel cell (MFC) anodic biofilm using Hungate roll-tube technique with iron oxide as indicator. At the same time, the dynamics of the microbial community structure was monitored during the pure culture isolation. The results show that maximum voltages of MFCs feeding with lactic acid, acetic acid and steroid wastewater are 0.57, 0.60 and 0.40 V respectively. The dominant bacteria isolated from seed sludge and anodic films feeding with acetate and lactate belong to phylum Proteobacteria; while steroid wastewater contains relative high diversity of bacteria, i. e. Proteobacteria, Firmicutes and Bacteroidetes. After enriching and culturing, two bacteria were consequently obtained, which shared the highest similarity with Enterobacter ludwigii and Citrobacter freundii respectively. When inoculated in MFC with lactic acid as the substrate, they produced maximum voltage of 0.10 and 0.17 V individually. This study shows that electrical active bacteria can be isolated from the MFC anodic biofilm using anaerobic gradient dilution culture techniques with iron oxide as indicator. Microbial community structure presents markedly shifting during the bacteria isolation owing to its selectivity.