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

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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