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

厌氧氨氧化污泥的批量制备方法及其细菌群落分析

基本信息

批准号：
12480165
负责人：
FURUKAWA Kenji
金额：
$ 6.66万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480165/
关键词：
anammox anaerobic ammonium oxidation nitrite nonwoven PVA gel bead bacterial community analysis Zoogloea ramigeera quantitative PCR analysis カラムリアクタ NH_4-N NO_2-N 窒素安定同位体 Planctomycete Anammox汚泥脱窒汚泥大量調製付着固定化担体

项目摘要

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

使用传统的悬浮生长方法来尝试驯化能够进行厌氧氨氧化（anammox）的生物体，但没有成功。因此，使用填充有新型聚酯非织造生物质载体的 1 L 上流式反应器进行了进一步的尝试，该载体先前已被证明可有效保留活性污泥。经过一年的连续驯化，成功开发了厌氧氨氧化培养物。随后，1 L 反应器的内容物被用作更大的 2 L 上流柱反应器的种子，该反应器也包含无纺布载体。在连续培养的一年内，一种独特的红色生物质（厌氧氨氧化生物体的特征）已遍布整个反应器。最后，将 2 L 反应器的驯化污泥用作扩大至 15 L 上流式反应器的种子，其中同样含有非织造材料。利用这个更大的反应器，成功建立了厌氧氨氧化培养物，并在连续运行三个月内实现了 0.5g N/L … More /d 的总氮去除活性。使用^15N标记的NH_4Cl，从NH_4^+和NO_2^-底物中释放出分子量为29的二氮气体，证实了厌氧氨氧化生物体常见的独特转化途径。此外，PVA凝胶珠被证明是厌氧氨氧化污泥的有效载体材料。使用装有 PVA 凝胶珠的上流式柱反应器进行厌氧氨氧化培养物的驯化，在连续培养六个月内达到 0.8 g N/L/d 的总氮去除活性。为了表征厌氧氨氧化污泥中细菌群落的组成，通过 PCR 扩增了 16S rDNA 序列，并使用 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细菌（菌株KSU-1）和Zoogloea ramigera 的存在。 KSU-1 占群落总种群的 72.8%，而动胶团属数量第二多，占 6%。这些结果与 FISH 分析结果非常一致。使用荧光成像，KSU-1 被证明形成包裹在一层薄薄的动胶团生物体中的球形簇。较少的