Development of Mass Preparation Method for Anammox Sludge and its Bacterial Community Analysis

使用传统的悬浮生长方法，尝试驯化能够进行厌氧氨氧化(Anammox)的微生物是不成功的。因此，使用1-L上流式反应器进行了进一步的尝试，该反应器填充了一种新型的聚酯非织造生物质载体，该载体以前被证明对活性污泥的保留是有效的。经过一年的连续驯化，成功地进行了厌氧氨氧化培养，将1-L反应器的内容物用作更大的2-L上流式反应器的种子，该反应器也包含非织造布载体。在连续培养的一年内，一种独特的红色生物量--厌氧氨氧化生物的特征--在整个反应器中蔓延开来。最后，用2-L反应器的驯化污泥作为种子放大到15-L上流式反应器，同样含有无纺布材料。在此较大的反应器中，成功地建立了厌氧氨氧化培养系统，总氮去除活性为0.5gN/L…连续运行3个月，日均可达3个以上。用~&lt；15&gt；N标记的NH_4CL从NH_4~+和NO_2~+底物中析出分子量为29的双氮气体，证实了厌氧氨氧化生物共有的独特转化途径。此外，PVA凝胶小球是厌氧氨氧化污泥的有效载体材料。采用填充PVA凝胶小球的上流式反应器对厌氧氨氧化污泥进行驯化，连续培养6个月后，总氮的去除活性达到0.8gN/L/d。证实存在一种以前未鉴定的厌氧氨氧化细菌(菌株KSU-1)和拉米格拉动物胶菌。KSU-1占群落总种群的72.8%，其次是动胶菌，占6%。这些结果与FISH分析结果吻合较好。利用荧光成像，KSU-1被证明形成了包裹在一层薄薄的动物胶生物体中的球形簇。较少

Attempts to acclimate organisms capable of anaerobic ammonium oxidation (anammox) were unsuccessful using traditional suspended growth methods. Further attempts were thus conducted using a 1-L up-flow colum reactor packed with a novel polyester nonwoven biomass carrier that was previously shown to be effective for retention of activated sludge. Following one year of continuous acclimation, an anammox culture was successfully developed Subsequently, the contents of the 1-L reactor were used as seed for a larger 2-L up-flow column reactor, also containing the nonwoven carrier. Within one year of continuous cultivation a distinctively red biomass, which is characteristic of anammox organisms, had spread throughout the reactor. Finally, the acclimated sludge of the 2-L reactor was used as seed for a scale-up to a 15-L up-flow reactor, again containing the nonwoven materials. With this larger reactor, an anammox culture was successfully established and a total-N removal activity of 0.5g N/L … More /d was achieved in three months of continuous operation. Using NH_4Cl labeled with ^<15>N, the evolution of di-nitrogen gas with a molecular weight of 29 from the NH_4^+ and NO_2^- substrates has served to confirm the unique pathway of transformation common to the anammox organisms.In addition, PVA gel beads were shown to be an effective carrier material for anammax sludge. Acclimation of an anammox culture was conducted using an up-flow column reactor packed with PVA gel beads and a total-N removal activity of 0.8 g N/L/d was attained within six months of continuous cultivation.To characterize the composition of the bacterial community in our anammox sludge, 16S rDNA sequences were amplified by PCR and comparative analyses using DNA databases were conducted. The presence of a previously unidentified anammox bacterium (strain KSU-1) and Zoogloea ramigera were confirmed. KSU-1 comprised 72.8% of the total community population and the Zoogloea species was the second most numerous at 6%. These results were in good agreement with results of FISH analysis. Using fluorescence imaging, KSU-1 was shown to form spherical clusters wrapped in a thin layer of Zoogloea organisms. Less