Development of the monitoring method of methanogenic microorganisms

1.乙酸乙酯产甲烷微生物的检测以含乙酸盐的合成废水为唯一碳源，对驯化后的厌氧消化污泥进行了连续两次中温厌氧培养。两种栽培的稀释率分别为0.025 d^-1和0.6d^-1。发酵液的荧光原位杂交表明，微生物在两种培养方式中都属于古生代占优势的领域。用从两种发酵液中提取的DNA构建16S rDNA文库。对文库DNA序列的系统发育分析表明，被归入古细菌属的克隆与甲藻属和甲藻属有亲缘关系。在两种培养方式中，大部分细菌克隆都属于芽孢杆菌/梭状芽孢杆菌类群。在这两种培养中都没有与已知的醋酸盐降解型合养细菌相关的克隆。利用实时荧光定量聚合酶链式反应技术对g-…的数量进行了估计。更多的是甲烷藻和甲烷八叠球菌。在两种稀释率条件下，甲烷藻的种群数量基本相同。但是，当稀释度为0.6d^-1~(-1)时，甲烷八叠球菌的种群数量高于稀释度为0.025 d^~(-1)时的数量。以含有丙酸盐的合成废水为唯一碳源，对驯化后的厌氧消化污泥进行了中温厌氧连续培养。稀释倍数在0.01~0.10d^~(-1)~(-1)之间呈阶梯式变化。发酵液的荧光原位杂交表明，微生物在所有稀释率下都属于古生菌占优势的领域。属于微生物科的微生物在古生界中占主导地位。在0.08d^~(-1)~(-1)的高稀释度下，参与H_2/CO_2产甲烷的辅酶F_&lt；420~gt；的量大于0.01d^~(-1)~(-1)的低稀释率。这一结果表明，低稀释速率下H2/CO2产甲烷活性高于高稀释速率下的产甲烷活性。较少

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