Development of analytical technique of change in microbial population for remediation in the polluted soil

微生物在各种污染环境中发挥着重要的修复作用。为了优化污染场地上微生物的活性，必须明确微生物分解细菌的最佳条件。应明确特定细菌和微生物种群的行为。本研究的目的是开发一种分析技术，用于阐明特定细菌在自然水体和土壤中的行为和微生物群落结构。土壤中大量的胶体往往阻碍了分子遗传学方法在土壤细菌上的直接应用。因此，在分析细菌的遗传物质之前，通常需要从土壤中提取细菌细胞。为此，对萃取剂的选择进行了研究。在EXP-…中评价了用苯二酮法从土壤中提取细菌的效率和微生物种群的代表性。更多的装备。用CTC(5-氰基-2，3-二苯基四氮氯铵)还原和平板计数技术对土壤中呼吸活性细胞的比例进行了评价。萃取剂为1/15M磷酸二氢钠、0.03M草酸钠和0.03M焦磷酸钠。细菌的萃取率定义为萃取物中的苯醌含量与土壤中的含量之比。与其他两种萃取剂相比，焦磷酸钠对土壤提取物中细菌和呼吸活性细胞的萃取率最高。此外，还用FISH方法测定了焦磷酸钠提取液中变形杆菌和γ亚类的细胞数。几乎所有变形杆菌α，β和γ亚类的细菌都含有UQ-10、UQ-8、UQ-9。实验结果表明，变形杆菌β，γ与α-亚类的比例与UQ-8、UQ-9与UQ-10的比例具有非常相似的关系。结果表明，利用FISH技术可以对土壤样品中的特定细菌进行定量，另外，利用滩涂实验装置研究沉积物粒度对微生物生态系统和水质净化的影响，以及分析沉积物中微生物群落结构，可以利用对苯二酚的谱和CTC还原来阐明沉积物中的微生物群落结构。较少

Microorganisms perform an important role for remediation in various polluted environment. To optimize the activity of microorganisms on contaminated sites, the ability of microorganisms must be clarified on the optimal condition for decomposing bacteria. The behavior of specific bacteria and microbial population should be clarified. The objective of this study is to develop the analytical technique for clarification of the behavior of specific bacteria and microbial community structure in natural waters and soils.The direct application of molecular genetic methods on soil bacteria is often prevented by the large amounts of soil colloids. Hence, an extraction of bacteria cells from soil is usually required before analysis of their genetic material. For this reason, the selection of a suitable extractant was investigated. The extraction efficiency of bacteria and the representation of the microbial population extracted from soil by using quinone-profiling method were evaluated in the exp … More eriments. The fraction of the respiratory active cell extracted from soil was also evaluated by using CTC (5-cyano-2, 3-ditolyl tetorazolium chloride) reduction and plate count techniques. 1/15M Na-phosphate, 0.03M Na-oxalate and 0.03M Na-pyrophosphate were used as the extractants. The extraction efficiency of bacteria was defined as the ratio of quinone contents in the extract to that in soil. Compared with other two extractants, Na-pyrophosphate demonstrated the highest extraction efficiency of bacteria and respiratory active cells in the extract from soil. In addition, cell numbers of Proteobacteria, and γ-subclass in extraction with Na- pyrophosphate was measured by FISH. Almost all bacteria of Proteobacteria α, β and γ-subclass contain UQ-10, UQ-8, UQ-9, respectively. The experimental results showed that there was very similar relationship between ratio of Proteobacteria β, γ to α-subclasses and ratio of UQ-8, UQ-9 to UQ-10, respectively. It indicates that specific bacteria in soil sample can be quantified by FISH technique.In addition, the quinone profile and CTC reduction can be used to clarify the effect of particle size of sediment on microbial-ecosystem and water purification using an experimental facility of the tidal flat, and to analyze microbial community structure in sediment. Less