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应用实时 qPCR 方法测量厌氧消化过程中产甲烷菌浓度,作为沼气产生能力的指标。

Application of a real-time qPCR method to measure the methanogen concentration during anaerobic digestion as an indicator of biogas production capacity.

机构信息

Department of Public Health and Microbiology, University of the Study of Turin, Via Santena 5 bis, 10126 Turin, Italy.

出版信息

J Environ Manage. 2012 Nov 30;111:173-7. doi: 10.1016/j.jenvman.2012.07.021. Epub 2012 Aug 19.

DOI:10.1016/j.jenvman.2012.07.021
PMID:22910214
Abstract

Biogas is an energy source that is produced via the anaerobic digestion of various organic materials, including waste-water sludge and organic urban wastes. Among the microorganisms involved in digestion, methanogens are the major microbiological group responsible for methane production. To study the microbiological equilibrium in an anaerobic reactor, we detected the methanogen concentration during wet digestion processes fed with pre-treated urban organic waste and waste-water sludge. Two different pre-treatments were used in successive experimental digestions: pressure-extrusion and turbo-mixing. Chemical parameters were collected to describe the process and its production. The method used is based on real-time quantitative PCR (RT-qPCR) with the functional gene mcrA as target. First, we evaluated the validity of the analyses. Next, we applied this method to 50 digestate samples and then we performed a statistical analysis. A positive and significant correlation between the biogas production rate and methanogen abundance was observed (r = 0.579, p < 0.001). This correlation holds both when considering all of the collected data and when the two data sets are separated. The pressure-extrusion pre-treatment allowed to obtain the higher methane amount and also the higher methanogen presence (F = 41.190, p < 0.01). Moreover a higher mean methanogen concentration was observed for production rate above than of 0.6 m(3) biogas/kg TVS (F = 7.053; p < 0.05). The applied method is suitable to describe microbiome into the anaerobic reactor, moreover methanogen concentration may have potential for use as a digestion optimisation tool.

摘要

沼气是一种能源,通过各种有机物质(包括废水污泥和有机城市废物)的厌氧消化产生。在消化过程中涉及的微生物中,产甲烷菌是负责产生甲烷的主要微生物群。为了研究厌氧反应器中的微生物平衡,我们在使用预处理城市有机废物和废水污泥进行湿法消化过程中检测了产甲烷菌的浓度。在连续的实验消化中使用了两种不同的预处理方法:挤压和涡轮混合。收集化学参数来描述该过程及其产物。所使用的方法基于以功能基因 mcrA 为靶标的实时定量 PCR(RT-qPCR)。首先,我们评估了分析的有效性。然后,我们将该方法应用于 50 个消化物样本,然后进行了统计分析。沼气产生率与产甲烷菌丰度之间存在正相关(r = 0.579,p < 0.001)。这一相关性在考虑所有收集的数据时以及在将两个数据集分开时都成立。挤压预处理允许获得更高的甲烷量和更高的产甲烷菌存在(F = 41.190,p < 0.01)。此外,对于高于 0.6 m(3)沼气/kg TVS 的产率,观察到更高的平均产甲烷菌浓度(F = 7.053;p < 0.05)。所应用的方法适用于描述厌氧反应器中的微生物组,并且产甲烷菌浓度可能有潜力用作消化优化工具。

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