Abstract

This study investigated the relationship between the temperature (35, 42, and 55 °C) used in temperature-phased anaerobic digestion (TPAD) and fate of methanogens between the two anaerobic digestion (AD) phases. Methanogens were profiled by using next generation sequencing (NGS) and droplet digital PCR approaches. The results showed that optimal combined temperatures for methane production were 55 °C during biological hydrolysis (BH) and 35 or 42 °C during AD. BH exhibited much lower archaeal population and was more susceptible to changes in temperature, compared to the AD phase. Additionally, we demonstrated, for the first time, that the BH step could affect the subsequent AD phase by altering AD methanogen composition and improve the stability of the process by enriching the rapidly growing Methanosarcina in the BH-AD process. These results are significant for understanding the mechanisms and stability of methane production in TPAD systems.

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