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

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 m3 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.

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► Waste water sludge and organic urban wastes are co-digested in pilot reactors.
► Two different waste pre-treatments are tested.
► Bio-molecular methanogen determination correlates with biogas yield in the reactor.
► Methanogen load can express biogas yield even after different input pre-treatment.