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[Display omitted] • DOC was the foremost parameter influencing the phytotoxicity of LD. • Temperature accelerated the stability and phytotoxicity disappearance of LD. • Open storage of LD at 30 °C for 60 days and 20 °C for 90 days of LD was safe. • Hermetic storage of LD at 30 °C for 120 days and 20 °C for 150 days of LD was safe. • For storage at 10 °C for 180 days, additional post-treatment is required. Identifying the stability and phytotoxicity of liquid digestate (LD) is necessary for safe agricultural utilization. Storage temperature, method, and time are critical factors that affect the stability and phytotoxicity of LD. This study therefore aimed to explore the dynamics of stability, phytotoxicity, and microbial community of LD in cattle farms under different storage conditions. The results showed that the contents of solids, organic matter, nitrogen, and phosphorous decreased during storage and exhibited temperature dependency. Conversely, the seed germination index increased, which was negatively correlated with dissolved organic carbon and ammonium nitrogen and positively correlated with certain bacteria (Thermovirga and Fastidiosipila). Open storage and/or higher temperature were found to contribute to the stabilization efficiency and phytotoxicity disappearance of LD. Open storage of LD at 30 °C for 60 days and 20 °C for 90 days was safe for its agricultural utilization, while hermetic storage of LD at 30 °C for 120 days and 20 °C for 150 days was safe. However, for storage at 10 °C for 180 days, additional post-treatment is required. [ABSTRACT FROM AUTHOR]

[Display omitted] • The effect of temperature rises and falls on anaerobic digestion was assessed. • 45 °C exhibits optimal methanogenic performance during temperature rises and falls. • An effective equilibrium between hydrolysis and methanation is achieved at 45 °C. • The shortest recovery time was 2 days following temperature fluctuation. This study implemented five successive temperature stages (35 °C–45 °C–55 °C–45 °C–35 °C) in a continuous stirred tank reactor, designated as D35-1, D45-1, D55, D45-2, and D35-2, respectively. The objective was to probe the mechanism of temperature transformations on methanogenesis performance, microbial community, and metabolic pathways. The suitability of 45 °C for anaerobic digestion (AD) under varying temperatures was emphatically expounded. Intriguingly, optimal methanogenesis performance was attained at 45 °C, with methane production reaching 238.51 mL/g VS/d in D45-1 and 245.91 mL/g VS/d in D45-2, signifying increases of 17.66% and 21.32% contrasted with D35-2, respectively. Additionally, considerable resistance to temperature shocks at 45 °C was also manifested, as evidenced by a recovery time of 2 days in D45-2. At 45 °C, an effective balance between hydrolysis and methanogenesis was observed, maintaining a dynamic equilibrium between acetoclastic and hydrogenotrophic methanogenesis. The primary drivers in D45-1 were Caldicoprobacter , Bathyarchaeia , and Methanosarcina , while Anaerolinea and Methanosarcina were the key determinants in D45-2. By contrast, methanogenesis was suppressed at other stages. Therefore, 45 °C emerges as a viable alternative to conventional temperatures when confronted with temperature fluctuations to optimize energy efficiency. The results provide novel insights into AD temperature management and pave the way for engineering applications. [ABSTRACT FROM AUTHOR]

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 °C, 45 °C, and 55 °C) and HRT (30, 25, and 20 d) for 116 days. The results demonstrated that the digester conducted at 45 °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 °C and 55 °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 °C, probably improving methane yield. These findings provide new insights into "short-circuit" mitigation and strategies to strengthen methane generation from biomass. • The digester at 45 °C exhibited a higher methane yield than those at 35 °C and 55 °C. • The optimal HRT for the 35 °C, 45 °C, and 55 °C digesters were 25, 30, and 30 d, respectively. • Temperature showed more influence on microbial regulation than HRT. • The enhanced acetoclastic methanogenesis path exhibited at 45 °C. [ABSTRACT FROM AUTHOR]