Mitigating short-circuits through synergistic temperature and hydraulic retention time control for enhancing methane yield in continuous stirred-tank reactors

Temperature and hydraulic retention time (HRT) control are crucial for mitigating the "short-circuit" issue and enhancing methane production in continuous stirred-tank reactors. This study focuses on the combined effects of temperature and HRT on methanation via operating the reactors at different temperatures (35 degrees C, 45 degrees C, and 55 degrees C) and HRT (30, 25, and 20 d) for 116 days. The results demonstrated that the digester conducted at 45 degrees C and an HRT of 30 d unexpectedly yielded the maximum average methane production of 296.08 ml/g volatile solids (VS). In contrast, the optimal HRT for the 35 degrees C and 55 degrees C digesters were 25 and 30 d, with a methane yield of 241.88 ml/g VS and 240.9 ml/g VS, respectively. High-throughput sequencing shows that Firmicutes, Bacteroidetes, Themotogae, Chloroflexi, and Acidobacteria dominated 90-98 % of the total bacteria. Temperature exhibited a significant influence on shaping microbial ecosystems, indicating that temperature, rather than HRT, is the primary factor to consider. Additionally, metagenomic analysis reveals that the levels of genes involved in acetoclastic and hydrogenotrophic methanogenic pathways were enhanced at 45 degrees C, probably improving methane yield. These findings provide new insights into "short-circuit" mitigation and strategies to strengthen methane generation from biomass.