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Within the European circular economy roadmap, it is important for wastewater treatment plant (WWTPs) to recover energy and become energy-neutral or -positive. In the last few years, it has become increasingly interesting to boost energy recovery through the biogas upgrading. The aim of this work is to study a rapid hydrogenotrophic methanogenic culture enrichment strategy capable of limiting the organic degradation unbalance and allowing a fast start-up phase of the in situ biogas upgrading reactors, at pilot- or full-scale. The approach was tested with two, plus one control, laboratory-scale continuous stirred tank reactors filled with anaerobic sludge collected from a full-scale WWTP. The experimentation lasted 50 days and was divided into five phases: the anaerobic digestion start-up followed by four H2 injection phases (H2/CO2 ranging from 1:1 to 4:1 on molar basis). Despite a temporary slight increase in the total concentration of volatile fatty acids during phase II (2.56 gCH3COOH·L−1), and in phase III a mild pH increase (anyway, below 7.4) indicating the expected CO2 depletion, the strategy proposed was effective. In the last phase, in the biogas a methane content of about 80% was achieved, thus suggesting that the use of H2/CO2 above the stoichiometric value could further improve the biological biogas upgrading.

The effect of the amount of hydrogen supplied for the in situ biological biogas upgrading was investigated by monitoring the process and evolution of the microbial community. Two parallel reactors, operated at 37°C for 211 days, were continuously fed with sewage sludge at a constant organic loading rate of 1.5 gCOD∙(L∙d)-1 and hydrogen (H2). The molar ratio of H2/CO2 was progressively increased from 0.5 : 1 to 7 : 1 to convert carbon dioxide (CO2) into biomethane via hydrogenotrophic methanogenesis. Changes in the biogas composition become statistically different above the stoichiometric H2/CO2 ratio (4 : 1). At a H2/CO2 ratio of 7 : 1, the methane content in the biogas reached 90%, without adversely affecting degradation of the organic matter. The possibility of selecting, adapting, and enriching the original biomass with target-oriented microorganisms able to biologically convert CO2 into methane was verified: high throughput sequencing of 16S rRNA gene revealed that hydrogenotrophic methanogens, belonging to Methanolinea and Methanobacterium genera, were dominant. Based on the outcomes of this study, further optimization and engineering of this process is feasible and needed as a means to boost energy recovery from sludge treatment.

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