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Anaerobic digestion has emerged as the most appealing waste management strategy in biorefineries. Particularly, recent studies have highlighted the energy advantages of waste co-digestion in industrial biorefineries and the use of two-stage systems. However, there are some concerns about moving the system from laboratory testing to industrial scale. One of them is the high level of investment that is required. Therefore, this study carried out a techno-economic analysis (scale-up and energy production, economic and risk analysis, and factorial design) to assess the feasibility of single- and two-stage systems in the treatment of cheese whey and glycerin for the production of hydrogen and methane. Scenarios (S1 to S9) considered thermophilic and mesophilic single and two-stage systems with different applied organic loading rates (OLR A ). The analyses of scale-up and energy production revealed that S3 (a thermophilic single-stage system operated at high OLR A 17.3 kg-COD.m -3 .d -1 ) and S9 (a thermophilic-mesophilic two-stage system operated at high OLR A 134.8 kg-COD.m -3 .d -1 and 20.5 kg-COD.m -3 .d -1 , respectively) were more compact and required lower initial investment compared to other scenarios. The risk analysis performed by a Monte Carlo simulation showed low investment risks (10 and 11%) for S3 and S9, respectively, being the electricity sales price, the key determining factor to define whether the project in the baseline scenario will result in profit or loss. Lastly, the factorial design revealed that while the net present value (NPV) is positively impacted by rising inflation and electricity sales price, it is negatively impacted by rising capitalization rate. Such assessments assist in making decisions regarding which system can be fully implemented, the best market circumstances for the investment, and how market changes may favorably or unfavorably affect the NPV and the internal rate of return (IRR)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Anaerobic digestion for CH 4 recovery in wastewater treatment has been carried out with different strategies to increase process efficiency, among which co-digestion and the two-stage process can be highlighted. In this context, this study aimed at evaluating the co-digestion of cheese whey and glycerol in a two-stage process using fluidized bed reactors, verifying the effect of increasing the organic loading rate (OLR) (2-20 g-COD.L -1 .d -1 ) and temperature (thermophilic and mesophilic) in the second stage methanogenic reactor. The mesophilic methanogenic reactor (R-Meso) (mean temperature of 22 °C) was more tolerant to high OLR and its best performance was at 20 g-COD.L -1 .d -1 , resulting in methane yield (MY) and methane production (MPR) of 273 mL-CH 4 .g-COD -1 and 5.8 L-CH 4 .L -1 .d -1 (with 67% of CH 4 ), respectively. Through 16S rRNA gene massive sequencing analysis, a greater diversity of microorganisms was identified in R-Meso than in R-Thermo (second stage methanogenic reactor, 55 °C). Firmicutes was the phyla with higher relative abundance in R-Thermo, while in R-Meso the most abundant ones were Proteobacteria and Bacteroidetes. Regarding the Archaea domain, a predominance of hydrogenotrophic microorganisms could be observed, being the genera Methanothermobacter and Methanobacterium the most abundant in R-Thermo and R-Meso, respectively. The two-stage system composed with a thermophilic acidogenic reactor + R-Meso was more adequate for the co-digestion of cheese whey and glycerol than the single-stage process, promoting increases of up to 47% in the energetic yield (10.3 kJ.kg-COD -1 ) and 14% in organic matter removal (90.5%)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)