Temperature plays a critical role in performance and stability of anaerobic digestion processes, subject to frequent meteorological fluctuations. However, state-of-the-art modeling and process control approaches for anaerobic digestion often do not consider the temporal dynamics of the temperature, which can influence microbial communities, kinetics, and chemical equilibrium, and consequently, biogas production efficiency. Therefore, to account for anaerobic digesters operating under fluctuating meteorological conditions, the Anaerobic Digestion Model no. 1 (ADM1) is mechanistically extended in this paper to incorporate temporal changes into temperature-dependent parameters by defining inhibition functions for microbial activities using the cardinal temperature model, and accounting for the lag in microbial adaptation to temperature fluctuations using a time-lag adaptation function. Thereafter, given that temperature fluctuations are a significant disturbance, a control framework based on Model Predictive Control (MPC) is developed to regulate the feeding flow rate and to ensure stable production rates despite temperature disturbances without relying on direct temperature control. An adaptive MPC approach is formulated based on a linear input–output model, where the parameters of the linear model are updated online to capture the nonlinear dynamics of the process and frequent changes in the dynamics accurately. In addition, a fuzzy logic system is employed to assign a reference trajectory for the production rate based on the temperature and its rate of change. Integrating this fuzzy logic system with the MPC controller enhances the production rate on warm days and avoids the operational failure in production on cold days. Additionally, to enhance biogas production rates, the feasibility of utilizing a portion of the produced biogas for external heating purposes is also investigated. It is demonstrated that by utilizing the proposed MPC approach, the additional amount of feed for the digester to produce methane required for a self-consumption biogas-fueled heating system can be calculated according to the meteorological variations. This enhances the process performance and stability. Finally, a thermally optimized dome digester semi-buried in the ground, operating under climate conditions of The Netherlands is considered as a case study to validate the extended model in agreement with biological and physicochemical behaviors of real-world applications, and to demonstrate the effectiveness of the proposed control system in handling temperature changes and enhancing performance.

Background: Elevated CO₂ partial pressure (pCO₂) has been proposed as a potential steering parameter for selective carboxylate production in mixed culture fermentation. It is anticipated that intermediate product spectrum and production rates, as well as changes in the microbial community, are (in)directly influenced by elevated pCO₂. However, it remains unclear how pCO₂ interacts with other operational conditions, namely substrate specificity, substrate-to-biomass (S/X) ratio and the presence of an additional electron donor, and what effect pCO₂ has on the exact composition of fermentation products. Here, we investigated possible steering effects of elevated pCO₂ combined with (1) mixed substrate (glycerol/glucose) provision; (2) subsequent increments in substrate concentration to increase the S/X ratio; and (3) formate as an additional electron donor. Results: Metabolite predominance, e.g., propionate vs. butyrate/acetate, and cell density, depended on interaction effects between pCO₂–S/X ratio and pCO₂–formate. Individual substrate consumption rates were negatively impacted by the interaction effect between pCO₂–S/X ratio and were not re-established after lowering the S/X ratio and adding formate. The product spectrum was influenced by the microbial community composition, which in turn, was modified by substrate type and the interaction effect between pCO₂–formate. High propionate and butyrate levels strongly correlated with Negativicutes and Clostridia predominance, respectively. After subsequent pressurized fermentation phases, the interaction effect between pCO₂–formate enabled a shift from propionate towards succinate production when mixed substrate was provided. Conclusions: Overall, interaction effects between elevated pCO₂, substrate specificity, high S/X ratio and availability of reducing equivalents from formate, rather than an isolated pCO₂ effect, modified the proportionality of propionate, butyrate and acetate in pressurized mixed substrate fermentations at the expense of reduced consumption rates and increased lag-phases. The interaction effect between elevated pCO₂ and formate was beneficial for succinate production and biomass growth with a glycerol/glucose mixture as the substrate. The positive effect may be attributed to the availability of extra reducing equivalents, likely enhanced carbon fixating activity and hindered propionate conversion due to increased concentration of undissociated carboxylic acids.

Background: Pressurised anaerobic digestion allows the production of biogas with a high content of methane and, at the same time, avoid the energy costs for the biogas upgrading and injection into the distribution grid. The technology carries potential, but the research faces practical constraints by a.o. the capital investment needed in high-pressure reactors and sensors and associated sampling limitations. In this work, the kinetic model of an autogenerative high-pressure anaerobic digestion of acetate, as the representative compound of the aceticlastic methanogenesis route, in batch configuration, is proposed to predict the dynamic performance of pressurised digesters and support future experimental work. The modelling of autogenerative high-pressure anaerobic digestion in batch configuration, which is not extensively studied and simulated in the present literature, was developed, calibrated, and validated by using experimental results available from the literature. Results: Under high-pressure conditions, the assessment of the Monod maximum specific uptake rate, the half-saturation constant and the first-order decay rate was carried out, and the values of 5.9 kg COD kg COD⁻¹ d⁻¹, 0.05 kg COD m⁻³ and 0.02 d⁻¹ were determined, respectively. By using the predicted values, excellent fittings of the final pressure, the CH₄ molar fraction and the specific methanogenic yield calculation were obtained. Likewise, the variation in the gas–liquid mass transfer coefficient by several orders of magnitude showed negligible effects on the model predictive values in terms of methane molar fraction of the produced biogas, while the final pressure seemed to be slightly influenced. Conclusions: The proposed model allowed to estimate the Monod maximum specific uptake rate for acetate, the half-saturation rate for acetate and the first-order decay rate constant, which were comparable with literature values reported for well-studied methanogens under anaerobic digestion at atmospheric pressure. The methane molar fraction and the final pressure predicted by the model showed different responses towards the variation of the gas–liquid mass transfer coefficient since the former seemed not to be affected by the variation of the gas–liquid mass transfer coefficient; in contrast, the final pressure seemed to be slightly influenced. The proposed approach may also allow to potentially identify the methanogens species able to be predominant at high pressure.

Fermentation at elevated hydrostatic pressure is a novel strategy targeting product selectivity. However, the role of inoculum history and cross-resistance, that is, acquired tolerance from incubation under distinctive environmental stress, remains unclear in high-pressure operation. In our here presented work, we studied fermentation and microbial community responses of halotolerant marine sediment inoculum (MSI) and anaerobic digester inoculum (ADI), pre-incubated in serum bottles at different temperatures and subsequently exposed to mild hydrostatic pressure (MHP; < 10 MPa) in stainless steel reactors. Results showed that MHP effects on microbial growth, activity, and community structure were strongly temperature-dependent. At moderate temperature (20°C), biomass yield and fermentation were not limited by MHP; suggesting a cross-resistance effect from incubation temperature and halotolerance. Low temperatures (10°C) and MHP imposed kinetic and bioenergetic limitations, constraining growth and product formation. Fermentation remained favorable in MSI at 28°C and ADI at 37°C, despite reduced biomass yield resulting from maintenance and decay proportionally increasing with temperature. Microbial community structure was modified by temperature during the enrichment, and slight differences observed after MHP-exposure did not compromise functionality. Results showed that the relation incubation temperature—halotolerance proved to be a modifier of microbial responses to MHP and could be potentially exploited in fermentations to modulate product/biomass ratio.

Small-scale electrical power generation (<100 kW) from biogas plants to provide off-grid electricity is of growing interest. Currently, gas engines are used to meet this demand. Alternatively, more efficient small-scale solid oxide fuel cells (SOFCs) can be used to enhance electricity generation from small-scale biogas plants. Most electricity generators require a constant gas supply and high gas quality in terms of absence of impurities like H ₂S. Therefore, to efficiently use the biogas from existing decentralized anaerobic digesters for electricity production, higher quality and stable biogas flow must be guaranteed. The installation of a biogas upgrading and buffer system could be considered; however, the cost implication could be high at a small scale as compared to locally available alterna-tives such as co-digestion and improved digester operation. Therefore, this study initially describes relevant literature related to feedstock pre-treatment, co-digestion and user operational practices of small-scale digesters, which theoretically could lead to major improvements of anaerobic digestion process efficiency. The theoretical preamble is then coupled to the results of a field study, which demonstrated that many locally available resources and user practices constitute frugal innovations with potential to improve biogas quality and digester performance in off-grid settings.

Volatile fatty acid accumulation is a sign of digester perturbation. Previous work showed the thermodynamic limitations of hydrogen and CO₂ in syntrophic propionate oxidation under elevated partial pressure of CO₂ (pCO₂). Here we study the effect of directional selection under increasing substrate load as a strategy to restructure the microbial community and induce cross-protection mechanisms to improve glucose and glycerol conversion performance under elevated pCO₂. After an adaptive laboratory evolution (ALE) process, viable cell density increased and predominant microbial groups were modified: an increase in Methanosaeta and syntrophic propionate oxidizing bacteria (SPOB) associated with the Smithella genus was found with glycerol as the substrate. A modest increase in SPOB along with a shift in the predominance of Methanobacterium toward Methanosaeta was observed with glucose as the substrate. The evolved inoculum showed affected diversity within archaeal spp. under 5 bar initial pCO₂; however, higher CH₄ yield resulted from enhanced propionate conversion linked to the community shifts and biomass adaptation during the ALE process. Moreover, the evolved inoculum attained increased cell viability with glucose and a marginal decrease with glycerol as the substrate. Results showed differences in terms of carbon flux distribution using the evolved inoculum under elevated pCO₂: glucose conversion resulted in a higher cell density and viability, whereas glycerol conversion led to higher propionate production whose enabled conversion reflected in increased CH₄ yield. Our results highlight that limited propionate conversion at elevated pCO₂ resulted from decreased cell viability and low abundance of syntrophic partners. This limitation can be mitigated by promoting alternative and more resilient SPOB and building up biomass adaptation to environmental conditions via directional selection of microbial community.

Simultaneous digestion and in situ biogas upgrading in high-pressure bioreactors will result in elevated CO2 partial pressure (pCO2). With the concomitant increase in dissolved CO2, microbial conversion processes may be affected beyond the impact of increased acidity. Elevated pCO2 was reported to affect the kinetics and thermodynamics of biochemical conversions because CO2 is an intermediate and end-product of the digestion process and modifies the carbonate equilibrium. Our results showed that increasing pCO2 from 0.3 to 8 bar in lab-scale batch reactors decreased the maximum substrate utilization rate (rsmax) for both syntrophic propionate and butyrate oxidation. These kinetic limitations are linked to an increased overall Gibbs free energy change (ΔGOverall) and a potential biochemical energy redistribution among syntrophic partners, which showed interdependence with hydrogen partial pressure (pH2). The bioenergetics analysis identified a moderate, direct impact of elevated pCO2 on propionate oxidation and a pH-mediated effect on butyrate oxidation. These constraints, combined with physiological limitations on growth exerted by increased acidity and inhibition due to higher concentrations of undissociated volatile fatty acids, help to explain the observed phenomena. Overall, this investigation sheds light on the role of elevated pCO2 in delicate biochemical syntrophic conversions by connecting kinetic, bioenergetic, and physiological effects.