Trace elements (TEs) are vital for anaerobic digestion (AD), due to their role as cofactors in many key enzymes. The aim of this study was to evaluate the effects of specific TE deficiencies on mixed microbial communities during AD of soluble polymer-free substrates, thus focusing on AD after hydrolysis. Three mesophilic (37 °C) continuous stirred-tank biogas reactors were depleted either of Co, Ni, or a combination of Se and W, respectively, by discontinuing their supplementation. Ni and Se/W depletion led to changes in methane kinetics, linked to progressive volatile fatty acid (VFA) accumulation, eventually resulting in process failure. No significant changes occurred in the Co-depleted reactor, indicating that the amount of Co present in the substrate in absence of supplementation was sufficient to maintain process stability. Archaeal communities remained fairly stable independent of TE concentrations, while bacterial communities gradually changed with VFA accumulation in Ni- and Se-/W-depleted reactors. Despite this, the communities remained relatively similar between these two reactors, suggesting that the major shifts in composition likely occurred due to the accumulating VFAs. Overall, the results indicate that Ni and Se/W depletion primarily lead to slower metabolic activities of methanogenic archaea and their syntrophic partners, which then has a ripple effect throughout the microbial community due to a gradual accumulation of intermediate fermentation products.

Sweden has the ambition to increase its annual biogas production from the current level of 1.9 to 15 TWh by 2030. The unused capacity of existing anaerobic digesters at wastewater treatment plants is among the options to accomplish this goal. This study investigated the feasibility of utilizing the organic fraction of municipal solid waste (OFMSW) as a co-substrate, with primary and waste-activated sewage sludge (PWASS) for production of biogas, corresponding to 3:1 ratio on volatile solid (VS) basis. The results demonstrated that co-digestion of OFMSW with PWASS at an organic loading rate of 5 gVS l⁻¹ day⁻¹ has the potential to increase the biogas production approximately four times. The daily biogas production increased from 1.0 ± 0.1 to 3.8 ± 0.3 l biogas l⁻¹ day⁻¹, corresponding to a specific methane production of 420 ± 30 Nml methane gVS⁻¹ during the laboratory experiment. Co-digestion of OFMSW with PWASS showed a 50:50 distribution of hydrogenotrophic and aceticlastic methanogens in the digester and enhanced the turnover kinetics of intermediate products (acetate, propionate, and oleate). Practical limitations potentially include the need for sludge dewatering to maintain a sufficient hydraulic retention time (17 days in this study), as well as additional energy consumption for mixing due to an increased sludge apparent viscosity (from 1.8 ± 0.1 to 45 ± 4.8 mPa*s in this study) at elevated OFMSW-loading rates.

Knowledge of microbial community dynamics in relation to process perturbations is fundamental to understand and deal with the instability of anaerobic digestion (AD) processes. This study aims to investigate the microbial community structure and function of a thermophilic AD process, fed with a chemically defined substrate, and its association with process performance stability. Next generation amplicon sequencing of 16S ribosomal RNA (rRNA) genes revealed that variations in relative abundances of the predominant bacterial species, Defluviitoga tunisiensis and Anaerobaculum hydrogeniformans, were not linked to the process performance stability, while dynamics of bacterial genera of low abundance, Coprothermobacter and Defluviitoga (other than D. tunisiensis), were associated with microbial community function and process stability. A decrease in the diversity of the archaeal community was observed in conjunction with process recovery and stable performance, implying that the high abundance of specific archaeal group(s) contributed to the stable AD. Dominance of hydrogenotrophic Methanoculleus particularly corresponded to an enhanced microbial acetate and propionate turnover capacity, whereas the prevalence of hydrogenotrophic Methanothermobacter and acetoclastic Methanosaeta was associated with instable AD. Acetate oxidation via syntrophic interactions between Coprothermobacter and Methanoculleus was potentially the main methane-formation pathway during the stable process. We observed that supplementation of Se and W to the medium improved the propionate turnover by the thermophilic consortium. The outcomes of our study provided insights into the community dynamics and trace element requirements in relation to the process performance stability of thermophilic AD.

Anaerobic digestion (AD) is an established process for integrating waste management with renewable energy and nutrient recovery. Much of the research in this field focuses on the utilisation of new substrates, yet their effects on operational aspects such as fluid behaviour and power requirement for mixing are commonly overlooked, despite their importance for process optimisation. This study analysed rheological characteristics of samples from 21 laboratory-scale continuous stirred-tank biogas reactors (CSTBRs) digesting a range of substrates, in order to evaluate substrate effect on mixing efficiency and power demand through computational fluid dynamics (CFD). The results show that substrate and process parameters, such as solids content and organic loading, all have a significant effect on CSTBR fluid rheology. The correlation levels between rheological and process parameters were different across substrates, while no specific fluid behaviour patterns could be associated with substrate choice. Substrate should thus be considered an equally important rheology effector as process parameters. Additional substrate-related parameters should be identified to explain the differences in correlations between rheological and process parameters across substrate groups. The CFD modelling revealed that the rheology differences among the AD processes have significant implications for mixing efficiency and power demand of the CSTBRs, highlighting the importance of considering the substrate-induced effects on CSTBR rheology before including a new substrate.