Development of Population Dynamics Analysis Methods of Soil Microbes for Optimizing Bioremediation of Contaminated Soil

开发土壤微生物种群动态分析方法以优化污染土壤的生物修复

• 批准号: 07458138
• 负责人: FUJIE Koichi
• 金额: $ 3.71万
• 依托单位: Toyohashi University of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07458138/
• 关键词: Bioremediation; Population dynamics; Quinone profile; PCR; Pentachlorophenol; Non-thermal discharge plasma; Sterilization; Cell condition; 土壌殺菌剤; OHラジカル; 混合培養系; 微生物群集構造; レーザトラップ; 静電配向力; マイクロマニピュレーション

In the present study, microbial quinones, genes and dielectric characteristics of microbial cells were introduced as indices for analyzing population dynamics of soil microbes for optimizing bioremediation of contaminated soil.Analysis method of respiratory quinones of soil microbes was developed, and the quinone profile defined as fractional content of each quinone specie was successfully used as a tool to analyze the change of soil microbial phase. The effect of bactericide such as pentachlorophenol on microbial phase of soil was investigated by using the method of quinone profile developed in this study.PCR technique was introduced to analyze the population of specific bacterium in a mixed culture system. A linear correlation between the logarithm of amplification of 16S rRNA and PCR cycle was found, which showing that PCR technique could be used to quantify specific bacterium. Cutting and no-cutting patterns by restriction enzyme of 16S rRNA was also suggested to classify microbes … More in a mixed culture system of bacteria.It was demonstrated that Escherichia coli and Schizosaccharomyces pombe cells were trapped with YAG laser and rotated by a high frequency alternating electric fields. The living cells had a peak critical rotation at 1MHz, however dead cells showed different peak critical rotation frequencies, which depended on the process of killing. These characteristics suggest that the measurement of the peak critical rotation frequency reflects cell condition. Purification method based on gradient force was also studied. Comb like electrodes with 0.2mm gap were constructed. Sample solution containing Escherichia coli was treated, and removal efficiency achieved almost 100%.Application of non-thermal discharge plasma for sterilization of soil was also studied. Using O_2 or Ar as a carrier gas, H_2O_2 was nebulized and the gas was treated by pulsed discharge plasma generated under atmospheric pressure in a concentric cylindrical electrode energized with a sharp-rising square wave voltage. Exposure of the plasma excited gas reduced the survivability of Bacillus subtilis spores on a film of less than 10^<-6> after 60sec. Less

为优化污染土壤的生物修复，引入微生物醌类、微生物基因和微生物细胞介电特性作为土壤微生物种群动态分析的指标，建立了土壤微生物呼吸醌类分析方法，并成功地将土壤微生物醌类组分含量谱作为分析土壤微生物相变化的工具。本研究采用醌类化合物谱法研究了五氯酚等污染物对土壤微生物相的影响，并采用PCR技术分析了混合培养体系中特定细菌的种群数量。16 SrRNA扩增对数与PCR循环数呈线性相关，表明PCR技术可用于特定细菌的定量。16 SrRNA限制性内切酶切割和非切割模式也可用于微生物的分类 ...更多信息 在混合培养体系中，利用YAG激光器捕获大肠杆菌和粟酒裂殖酵母细胞，并在高频交变电场作用下旋转。活细胞在1 MHz处有一个临界旋转峰值，而死细胞则有不同的临界旋转峰值频率，这取决于细胞的杀伤过程。这些特征表明，峰值临界旋转频率的测量反映了细胞状况。研究了基于梯度力的提纯方法。梳状电极与0.2mm的间隙构造。对含大肠杆菌的样品溶液进行了处理，去除率接近100%，并研究了低温放电等离子体在土壤消毒中的应用。以O_2或Ar为载气，H_2O_2雾化后，在大气压下，在同心圆电极中产生脉冲放电等离子体，并施加陡升的方波电压。暴露于等离子体激发的气体降低了枯草芽孢杆菌孢子在60秒后小于10 μ g的膜上的存活率<-6>。少

项目成果

Tohru Mori, Koichi Fujie: "Accelerated Microbial Degradation of Chlorothalonil in Soils Receiving Farmyard Manure" J. Soil Science and Plant Nutrition. 42(2). 315-322 (1995)

Tohru Mori、Koichi Fujie：“接受农家肥的土壤中百菌清的加速微生物降解”J.土壤科学与植物营养。

浦野紘平,藤江幸一,他: "水道水のAmes変異原生に関する研究" 水環境学会誌. 18. 1001-10011 (1995)

Kohei Urano、Koichi Fujie 等人：“自来水中艾姆斯致突变性的研究”水环境学会杂志 18. 1001-10011 (1995)。

B.-R.Lim, X.Huang, H.-Y.Hu and K.Fujie: "Solid Phase Aerobic Digestion of High Strength Organic Wastewater using Adsorbent Polymer Gel" Water Science and Technology. (in press).

B.-R.Lim、X.Huang、H.-Y.Hu 和 K.Fujie：“利用吸附聚合物凝胶固相好氧消化高浓度有机废水”水科学与技术。

H.-Y.Hu, K.Fujie, H.Tanaka, T.Makabe and K.Urano: "Respiratory Quinone Profile as a Tool for Refractory Chemical Biodegradation Study" Water Science and Technology. (in press).

H.-Y.Hu、K.Fujie、H.Tanaka、T.Makabe 和 K.Urano：“呼吸醌谱作为难降解化学生物降解研究的工具”水科学与技术。

西岡将輝、平野 研: "細胞の回転操作および回転特性の評価" 静電気学会講演論文集'95. 425-428 (1995)

Masaki Nishioka，Ken Hirano：“细胞的旋转操作和旋转特性的评估”静电学会论文集95 425-428（1995）。