In this study, we investigated thermophilic (55 °C) anaerobic digestion (AD) performance and microbial community structure, before and after hydrogen addition, in a novel hybrid gas-stirred tank reactor (GSTR) implemented with a partial immobilization of the microbial community and fed with second cheese whey (SCW). The results showed that H₂ addition led to a 25% increase in the methane production rate and to a decrease of 13% in the CH₄ concentration as compared with the control. The recovery of methane content (56%) was reached by decreasing the H₂ flow rate. The microbial community investigations were performed on effluent (EF) and on interstitial matrix (IM) inside the immobilized area. Before H₂ addition, the Anaerobaculaceae (42%) and Lachnospiraceae (27%) families dominated among bacteria in the effluent, and the Thermodesulfobiaceae (32%) and Lachnospiraceae (30%) families dominated in the interstitial matrix. After H₂ addition, microbial abundance showed an increase in the bacteria and archaea communities in the interstitial matrix. The Thermodesulfobiaceae family (29%)remained dominant in the interstitial matrix, suggesting its crucial role in the immobilized community and the SHA-31 family was enriched in both the effluent (36%) and the interstitial matrix (15%). The predominance of archaea Methanothermobacter thermoautrophicus indicated that CH₄ was produced almost exclusively by the hydrogenotrophic pathway.

Some livestock farms rely on anaerobic digestion (AD) technology for manure disposal, thus obtaining energy (biogas) and fertilizer (digestate). Mixtures of antibiotics used for animal health often occur in organic waste and their possible synergistic/antagonistic effects on microorganisms involved in AD are still poorly studied. This work focuses on the effects of adding ciprofloxacin, alone (5 mg L⁻¹) and in combination with sulfamethoxazole (2.5–5–10 mg L⁻¹), on AD efficiency and microbial community structure. The experiment consisted of 90-day cattle manure batch tests and antibiotic removal percentages were assessed. Adding antibiotics always promoted CH₄ and H₂ production compared to untreated controls; however, CH₄ production was lowered with the highest ciprofloxacin (CIP) concentrations. The overall results show antibiotic degradation caused by acidogenic Bacteria, and CH₄ was mainly produced through the hydrogenotrophic-pathway by methanogenic Archaea. Shifts in microbial community abundance (DAPI counts) and composition (Illumina-MiSeq and FISH analyses) were observed.

Simultaneous bioproduction of hydrogen and ethanol from cheaper waste feedstock has the potential for the development of a more cost-effective biofuel generation process. Crude glycerol (CG), a by-product of the biodiesel industry, is a renewable resource, abundant, sold at low prices and available worldwide. However, the main CG limitations in fermentation processes are mainly related to the presence of impurities and the lack of nitrogen sources, both acting on microbial activity. In this study, a fermentation process with CG was improved using a highly specific microbial consortium called GlyCeroL (GCL). The process was developed in fed-batch fermentation mode using not diluted substrate and carried out under non-sterile conditions and at increasing amounts of the substrate (from 20 to 80 gL⁻¹ of glycerol). The results showed higher H₂ (from 6 to 8 LL⁻¹) and EtOH (from 13 to 20 gL⁻¹) production by increasing glycerol concentration from 20 to 40 gL⁻¹. On the other hand, a decrease in glycerol degradation efficiency (from 75 to 56%) was observed. Then, the nitrogen sparging strategy was applied. Using CG of 40 gL⁻¹, process improvement was achieved, leading to the increased production of hydrogen (10 LL⁻¹) but not that of ethanol (20 gL⁻¹). A further increase to 60 gL⁻¹ of glycerol produced a slight increment of EtOH (21 gL⁻¹) and H₂ (11 gL⁻¹) but a sharp decrease in glycerol degradation efficiency (41%). Acetate, as the main impurity of CG, was an additional carbon source for GCL microorganisms contributing to EtOH production and increasing that of lactic acid to restore the redox balance. The Denaturing Gradient Gel Electrophoresis (DGGE) fingerprint at the end of all fed-batch fermentations supported the robustness of GCL functional units and their adaptability to fermentation conditions.

This study evaluated different strategies to increase gas–liquid mass transfer in a randomly packed gas stirred tank reactor (GSTR) continuously fed with second cheese whey (SCW), at thermophilic condition (55 °C), for the purpose of carrying out in situ biogas upgrading. Two different H₂ addition rates (1.18 and 1.47 L_H2 L_R⁻¹ d⁻¹) and three different biogas recirculation rates (118, 176 and 235 L L_R⁻¹ d⁻¹) were applied. The higher recirculation rate showed the best upgrading performance; H₂ utilization efficiency averaged 88%, and the CH₄ concentration in biogas increased from 49.3% during conventional anaerobic digestion to 75%, with a methane evolution rate of 0.37 L_CH4 L_R⁻¹ d⁻¹. The microbial community samples were collected at the end of each experimental phase, as well as one of the thermophilic sludge used as inoculum; metanogenomic analysis was performed using Illumina-based 16S sequencing. The whole microbial community composition was kept quite stable throughout the conventional anaerobic digestion (AD) and during the H₂ addition experimental phases (UP1, UP2, UP3, UP4). On the contrary, the methanogens community was deeply modified by the addition of H₂ to the GSTR. Methanogens of the Methanoculleus genus progressively increased in UP1 (47%) and UP2 (51%) until they became dominant in UP3 (94%) and UP4 (77%). At the same time, members of Methanotermobacter genus decreased to 19%, 23%, 3% and 10% in UP1, UP2, UP3 and UP4, respectively. In addition, members of the Methanosarcina genus decreased during the hydrogen addition phases.