2001 Fiscal Year Final Research Report Summary
Development of the monitoring method of methanogenic microorganisms
| Project/Area Number |
12680573
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
環境保全
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| Research Institution | Kumamoto University |
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Principal Investigator |
SHIGEMATSU Toru Kumamoto University, Graduate School of Science and Technology, Assistant Professor, 大学院・自然科学研究科, 助手 (10315286)
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| Co-Investigator(Kenkyū-buntansha) |
KIDA Kenji Kumamoto University, Faculty of Engineering, Professor, 大学院・自然科学研究科, 助手 (00195306)
MORIMURA Shigeru Kumamoto University, Graduate School of Science and Technology, Associate Professor, 大学院・自然科学研究科, 助手 (20230146)
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| Project Period (FY) |
2000 – 2001
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| Keywords | Methane fermentation / Methanogen / Microbial flora / DNA probe / 16S rRNA / Wastewater treatment / Anaerobic treatment |
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| Research Abstract |
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
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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
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