Enhancement of Microbial Function for a Sulfur Redox Cycle Action in a Sewage Treatment Process Combining an UASB and an Aerobic Filter Reactor

结合 UASB 和好氧过滤反应器增强污水处理过程中硫氧化还原循环作用的微生物功能

• 批准号: 16560489
• 负责人: ARAKI Nobuo
• 金额: $ 2.37万
• 依托单位: Nagaoka National College of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16560489/
• 关键词: sewage treatment; sulfate-reducing bacteria; sulfur-oxidizing bacteria; microbial structure; real-time PCR assay; functional genes; 硫黄酸化細菌

The structure of microbial community was analyzed on sludge samples taken from the aerobic biofilter reactor by using clone libraries of 16S rRNA genes. Clone similar to Thiobacillus and Thiothrix was counted at 10 and 7, respectively, among 188 clones randomly selected from the libraries. Approximately 7% of DAPI stained total cells in the biofilter sludge hybridized with the probe Thio840 specific for Thiobacillus group. Real-time PCR using SYBR green I was employed with BONE663cF-Thio840R primer set to determine the copy number of 16S rRNA gene originated from Thiobacillus. High concentration of sulfide at 30 mg-S/L in the influent strongly enhanced abundance of the gene of 10^5 copies/ng levels in total DNA extracted from the aerobic biofilter sludge. the diversity of adenosine-5'-phosphosulfate reductase (apsA) genes, which produce a key enzyme for sulfate respiration and are present in all sulfate-respiring prokaryotes (SRPs), in UASB sludge was analyzed by cloning and sequencing techniques. The apsA amino acid sequence-based phylogenetic tree showed that the identified clones belonged to four groups : the sulfate-reducing bacterial group, Syntrophobacteraceae, Thiobacillus and the clone cluster group, distantly related to any previously described apsA clone. Real-time PCR quantification which used the primer sets specific for each of the four apsA gene sequences revealed that Desulfobulbaceae and Desulfovibrionales were the predominant species all of the sulfate-reducing bacteria in the UASB sludge throughout the experimental period. An excellent correlation of apsA gene abundance and behavior was found between the sulfate-reducing bacteria and Thiobacillus, as well as between Syntrophobacteraceae and the unknown apsA clone group.

利用 16S rRNA 基因克隆文库对取自好氧生物滤池反应器的污泥样品进行微生物群落结构分析。在从文库中随机选择的 188 个克隆中，与硫杆菌属和硫丝菌属相似的克隆分别计数为 10 个和 7 个。生物过滤器污泥中约 7% 的 DAPI 染色总细胞与硫杆菌属特异性探针 Thio840 杂交。使用 SYBR green I 的实时 PCR 和 BONE663cF-Thio840R 引物组来确定源自硫杆菌的 16S rRNA 基因的拷贝数。进水中 30 mg-S/L 的高浓度硫化物强烈增强了从好氧生物过滤器污泥中提取的总 DNA 中基因的丰度，达到 10^5 拷贝/ng 水平。通过克隆和测序技术分析了 UASB 污泥中 5'-磷酸腺苷还原酶 (apsA) 基因的多样性，该基因产生硫酸盐呼吸的关键酶，存在于所有硫酸盐呼吸原核生物 (SRP) 中。基于apsA氨基酸序列的系统发育树显示，鉴定的克隆属于四个类群：硫酸盐还原菌群、互养细菌科、硫杆菌属和克隆簇群，与任何先前描述的apsA克隆有远亲关系。使用四种 apsA 基因序列中每一种的特异性引物组进行的实时 PCR 定量显示，在整个实验期间，Desulfobulbaceae 和 Desulfovibrionales 是 UASB 污泥中所有硫酸盐还原菌的主要物种。在硫酸盐还原菌和硫杆菌之间，以及合成杆菌科和未知的 apsA 克隆群之间，发现 apsA 基因丰度和行为具有极好的相关性。

项目成果

Multiplex real-time PCR法を用いた硫黄酸化菌と硫酸塩還元菌の同時定量

使用多重实时 PCR 方法同时定量硫氧化菌和硫酸盐还原菌

• 发表时间: 2005
• 期刊: 環境工学研究論文集 42
• 作者: 川上周司;荒木信夫;山口隆司;角野晴彦;高橋優信 他4名
• 通讯作者: 高橋優信 他4名

Feasibility study of the pilot-scale sewage treatment system combining of UASB and aerated fixed bed reactor under ambient temperature

UASB与曝气固定床反应器常温污水处理中试系统可行性研究

• 发表时间: 2004
• 期刊: Environmental Engineering Research Vol.41
• 作者: H.SUMINO;M.TAKAHASHI;T.YAMAGUCHI;K.ABE;N.ARAKI;S.YAMAZAKI;S.SHIMOZAKI;A.NAGANO;N.NISHIO
• 通讯作者: N.NISHIO

硫黄酸化還元サイクルを活性化したパイロットスケールUAS8/DHSシステムによる都市下水の処理性能評価

使用带有激活硫氧化还原循环的中试规模 UAS8/DHS 系统评估城市污水处理性能

• 发表时间: 2005
• 期刊: 環境工学研究論文集 42
• 作者: 高橋優信;山口隆司;角野晴彦;荒木信夫;山崎慎一;西尾尚道
• 通讯作者: 西尾尚道

Diversity Analysis and Quantification of Functional Gene in Sulfate-reducing Bacteria in Novel Sewage Treatment System Enhancing Sulfur-redox Cycle Action

增强硫氧化还原循环作用的新型污水处理系统中硫酸盐还原菌功能基因的多样性分析和定量

• 发表时间: 2005
• 期刊: Journal of Japan Society on Water Environment Vol.28, No.8
• 作者: K.ABE;N.ARAKI;H.IMACHI;H.SUMINO;T.YAMAGUCHI;N.NISHIO
• 通讯作者: N.NISHIO

Simultaneous Quantification of Sulfur-Oxidizing Bacteria and Sulfate-Reducing Bacteria by Multiplex Real-time PCR

通过多重实时 PCR 同时定量硫氧化细菌和硫酸盐还原细菌

• 发表时间: 2005
• 期刊: Environmental Engineering Research. Vol. 42
• 作者: Shuji Kawasaki;Nobuo Araki;Takashi Yamaguchi;Haruhiko Sumino;Masanobu Takahashi Shinichi Yamazaki;Hiroyuki Imachi;Akiyoshi Ohashi;Hideki Harada
• 通讯作者: Hideki Harada