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

尝试驯化生物能够厌氧氨氧化（anammox）是不成功的使用传统的悬浮生长方法。因此，进一步的尝试是使用1升上流式柱反应器进行填充有一种新的聚酯非织造生物质载体，先前被证明是有效的活性污泥的保留。经过一年的连续驯化，厌氧氨氧化培养物被成功开发。随后，将1-L反应器的内容物用作更大的2-L上流式柱反应器的种子，该反应器也含有非织造载体。在连续培养的一年内，一种独特的红色生物质，这是厌氧氨氧化生物体的特征，已经遍布整个反应器。最后，将2-L反应器的驯化污泥用作种子，用于按比例放大至15-L上流式反应器，其也含有非织造材料。利用该反应器成功地建立了厌氧氨氧化培养体系，其总氮去除率可达0.5gN/L ...更多信息 /d在连续运行三个月内实现。用^ N标记NH_4Cl<15>，从NH_4^+和NO_2^-底物中释放出分子量为29的双氮气体，证实了厌氧氨氧化微生物的独特转化途径。厌氧氨氧化培养物的驯化使用的上流式柱反应器填充PVA凝胶珠和总氮去除活性达到0.8 g N/L/d 6个月的连续培养。为了表征我们的厌氧氨氧化污泥中的细菌群落组成，16 S rDNA序列的PCR扩增和DNA数据库进行了比较分析。证实了先前未鉴定的厌氧氨氧化菌（菌株KSU-1）和枝状动胶菌的存在。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分析结果吻合良好。使用荧光成像，KSU-1显示出形成包裹在一层薄薄的动胶菌有机体中的球形簇。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