生物转盘中厌氧氨氧化污泥特性及菌种的初步鉴定试验研究

Characteristics and preliminary identification of ANAMMOX bacteria in a rotating biological contactor

对厌氧生物转盘系统中具有厌氧氨氧化功能的污泥进行生物特性及分子生物学的初步鉴定试验研究.结果表明:污泥呈砖红色,系统中除附着在盘片上的生物膜外,槽间还存在颗粒污泥及少量絮体;镜检发现细菌主要为短杆状和球状两种形态,革兰氏阴性;SEM发现颗粒污泥多孔隙,颗粒和生物膜表面荚膜包围有球菌;间歇试验表明最大氨氧化速率为0.210 kg NH4+-N.kg-1VSS.d-1,总氮污泥去除负荷达0.311 kg TN.kg-1VSS.d-1;用厌氧氨氧化菌特异性引物进行PCR扩增,琼脂糖凝胶电泳显示清晰的特异性条带;核酸序列经测序及BLAST功能比对、进一步构建进化树表明该细菌与已报道的厌氧氨氧化菌Candidatus Kueneniastuttgartiensis同属,与其基因序列相似性为100%,初步鉴定了其种属.

Characteristics of ANAMMOX sludge in an anaerobic rotating biological contactor were studied and preliminarily analyzed using molecule biological methods. The color of the sludge is brick red. Besides biofilm attached on the disks, there existed flocs and granule with many pores in the tank. The shape of bacteria was discovered as rod shaped and globosely under microscope; with some coccus found gathered on the surface of both the granule sludge and biofilm through scanning electron microscope. TN removal loading and the max anaerobic ammonia oxidation rate were 0. 311 kg TN·kg^-1VSS·d^-1 and 0. 210 kg NH4^＋-N·kg^-1VSS·d^-1, respectively. Clear special bar was displayed in gelose gelatin electrophoresis after PCR amplification. It showed that 16SrRNA of the bacteria is 100% similar to Candidatus Kuenenia stuttgartiensis and same genus to it through the function of BLAST in GenBank and constructing phylogenetic tree, which testifies the genus of the bacteria.