Development of the monitoring method of methanogenic microorganisms

1.乙酸盐产甲烷微生物的检测以乙酸盐为唯一碳源的人工合成废水为驯化厌氧消化污泥，进行了两次连续中温厌氧培养。两种培养物的稀释率分别为0.025 d^<-1>和0.6 d^<-1>。荧光原位杂交的培养液显示，微生物分类的域degreea占主导地位，在这两个培养。使用从两种培养液中提取的DNA构建16 S rDNA文库。系统发育分析表明，该文库的DNA序列的克隆分类在结构域的相关属Methanosaeta和Methanosarcina。在两种培养条件下，绝大多数细菌克隆都属于芽孢杆菌/梭菌群。在两种培养物中均没有与已知的乙酸盐降解互养细菌相关的克隆。实时荧光定量PCR实验，以估计种群的g 关于我们 Enera Methanosaeta和Methanosarcina。在两种稀释率条件下，甲烷刺水蚤的种群数量基本相同。但甲烷八叠球菌在稀释率为0.6d ^时的种群数量<-1>高于稀释率为0.025d ^时的种群数量<-1>。以丙酸盐为唯一碳源，对驯化好的厌氧消化污泥进行中温厌氧连续培养。稀释率从0.01到0.10 d/L逐步变化<-1>。荧光原位杂交的培养液显示，微生物分类的域degreea占主导地位，在所有的稀释率。分类在甲烷微菌科的微生物在该领域占主导地位。在<420>高稀释率（0.08d ^）下，参与H_2/CO_2产甲烷反应的辅酶F_的量<-1>大于低稀释率（0.01d ^）下的辅酶F_的量<-1>。这一结果表明，在低稀释率下H_2/CO_2的产甲烷活性高于高稀释率。少

1. Detection of microorganisms involved in methanogenesis from acetate.Two continuous mesophilic anaerobic cultivations of acclimatized anaerobic digestion sludge with a synthetic wastewater containing acetate as a sole carbon source were carried out. The dilution rates of the two cultivation were 0.025 d^<-1> and 0.6 d^<-1>. Fluorescence in situ hybridization of the culture broths revealed that microorganisms classified the domain Archaea dominated in both cultivations. The 16S rDNA libraries were constructed using the extracted DNA from both culture broths. The phylogenetic analysis of the DNA sequences of the libraries showed that clones classified in the domain Archaea related to genera Methanosaeta and Methanosarcina. Most of bacterial-clones were classified in Bacillus/Clostridium group in both cultivations. No clones related to known acetate-degrading syntrophic bacteria in both cultivations. Real-time quantitative PCR experiments was carried out to estimate the populations of g … More enera Methanosaeta and Methanosarcina. The populations of Methanosaeta were almost same under the two dilution rates conditions. But, the population of Methanosarcina was higher at the dilution rate of 0.6 d^<-1> than that at a dilution rate of 0.025 d^<-1>.2. Detection of microorganisms involved in methanogenesis from propionate.Continuous mesophilic anaerobic cultivations of acclimatized anaerobic digestion sludge with a synthetic wastewater containing propionate as a sole carbon source was carried out. The dilution rate was varied stepwise from 0.01 to 0.10 d^<-1>. Fluorescence in situ hybridization of the culture broths revealed that microorganisms classified the domain Archaea dominated at all dilution rates. Microorganisms classified in the family Methanomicrobiaceae dominated in the domain Archaea. The amount of coenzyme F_<420>, which involves in methanogenesis from H_2/CO_2, was larger at a high dilution rate of 0.08 d^<-1> than that at a low dilution rate of 0.01 d^<-1>. This result suggests that the methanogenic activity from H_2/CO_2, was higher at the low dilution rate than that at the high dilution rate. Less