Effects of temperature and hydraulic retention time on acetotrophic pathways and performance in high-rate sludge digestion.

High-rate anaerobic digestion of organic solids requires rapid hydrolysis and enhanced methanogenic growth rates, which can be achieved through elevated temperature (>55 °C) at short hydraulic retention times (HRT). This study assesses the effect of temperatures between 55 °C and 65 °C and HRTs between 2 and 4 days on process performance, microbial community structure, microbial capability, and acetotrophic pathways in thermophilic anaerobic reactors. Increasing the temperature did not enhance volatile solids (VS) destruction above the base value of 37% achieved at 55 °C and 4 days HRT. Stable isotopic signatures (δ13C) revealed that elevated temperature promoted syntrophic acetate oxidation, which accounted for 60% of the methane formation at 55 °C, and increasing substantially to 100% at 65 °C. The acetate consumption capacity dropped with increasing temperature (from 0.69-0.81 gCOD gVS(-1) d(-1) at 55 °C to 0.21-0.35 gCOD gVS(-1) d(-1) at 65 °C), based on specific activity testing of reactor contents. Community analysis using 16S rRNA pyrosequencing revealed the dominance of Methanosarcina at 55-60 °C. However, a further increase to 65 °C resulted in loss of Methanosarcina, with an accumulation of organic acids and reduced methane production. Similar issues were observed when reducing the HRT to 2 days, indicating that temperature<60 °C and HRT>3 days are critical to operate these systems stably.