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This study aimed to characterize the prokaryotic community and putative microbial interactions involved in hydrogen (H 2 ) production during the dark fermentation (DF) process, applying principal components analysis (PCA) to correlate changes in operational, physicochemical, and biological variables. For this purpose, a continuous stirred-tank reactor-type digester fed with tequila vinasses was operated at 24, 18, and 12 h of hydraulic retention times (HRTs) to apply organic loading rates of 20, 36, and 54 g-COD L -1 d -1 , corresponding to stages I, II, and III, respectively. Results indicated high population dynamics for Archaea during the DF process toward a decrease in total sequences from 6299 to 99. Concerning the Bacteria community, lactic acid bacteria (LAB) were dominant reaching a relative abundance of 57.67%, while dominant H 2 -producing bacteria (HPB) decreased from 25.76% to 21.06% during stage III. Putative competitive exclusion mechanisms such as competition for substrates, bacteriocins production, and micronutrient depletion carried out by Archaea and non-H 2 -producing bacteria (non-HPB), especially LAB, could negatively impact the dominance of HPB such as Ethanoligenens harbinense and Clostridium tyrobutyricum. As a consequence, low maximal volumetric H 2 production rate (672 mL-H 2 L -1 d -1 ) and yield (3.88 mol-H 2 assimilated sugars -1 ) were obtained. The global scenario obtained by PCA correlations suggested that C. tyrobutyricum positively impacted H 2 molar yield through butyrate fermentation using the butyryl-CoA:acetate CoA transferase pathway, while the most abundant HPB E. harbinense decreased its relative abundance at the shortest HRT toward the dominance of non-HPB. This study provides new insights into the microbial interactions and helps to better understand the DF performance for H 2 production using tequila vinasses as substrate. KEY POINTS: • E. harbinense and C. tyrobutyricum were responsible for H 2 production. • Clostridiales used acetate and butyrate fermentations for H 2 production. • LAB won the competition for sugars against Clostridiales during DF. • Putative bacteriocins production and micronutrients depletion could favor LAB., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Aims: The purpose of this study was to apply cDNA approach for the characterization of active prokaryotic community to understand microbial scenarios and performance of an AnSBR digester fed with acid hydrolysates of Agave tequilana var. azul bagasse (ATAB)., Methods and Results: The digester was implemented for methane production under organic loading rate (OLR) disturbances to correlate physicochemical variables with changes in abundance, diversity and population dynamics of active Bacteria and Archaea by principal components analysis (PCA). Results indicated that methane yield increased as well as active syntrophic relationships for interspecies hydrogen/formate (Anaerolinaceae-Methanobacterium beijingense) and acetate (Anaerolinaceae-Methanosaeta concilii) transfers at 8 g-COD l -1  day -1 . However, methane yield was negatively affected at 16 g-COD l -1  day -1 due to the competition for acetate by active Desulfovibrio marrakechensis and volatile fatty acids inhibition., Conclusions: Microbial scenarios obtained by PCA correlations indicated that methane production from acid hydrolysates of ATAB was feasible at 8 g-COD l -1  day -1 . The digester operation at higher OLR only favoured methanogenesis by the hydrogenotrophic pathway., Significance and Impact of the Study: Only cDNA analysis showed Archaea population dynamics, exhibiting high correlation with physicochemical variables towards the understanding of the methanogenic digester performance during OLR disturbances., (© 2019 The Society for Applied Microbiology.)

A hybrid (continuous-discrete) cascade control is proposed to regulate both, volatile fatty acids (VFA) and chemical oxygen demand (COD) concentrations in two-stage (acidogenic-methanogenic) anaerobic digestion (TSAD) processes. The outer loop is a discrete controller that regulates the COD concentration of the methanogenic bioreactor by using a daily off-line measurement and that modifies the set-point tracked by inner loop, which manipulates the dilution rate to regulate the VFA concentration of the acidogenic bioreactor, estimated by continuous on-line conductivity measurements, avoiding acidification. The experimental validation was conducted in a TSAD process for the treatment of tequila vinasses during 110days. Results showed that the proposed cascade control scheme was able to achieve the VFA and COD regulation by using conventional measurements under different set-point values in spite of adverse common scenarios in full-scale anaerobic digestion processes. Microbial composition analysis showed that the controller also favors the abundance and diversity toward methane production., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

An artificial neural network (ANN) was implemented to model the light profile pattern inside a photobioreactor (PBR) that uses a toroidal light arrangement. The PBR uses Tequila vinasses as culture medium and purple non-sulfur bacteria Rhodopseudomonas palustris as biocatalyzer. The performance of the ANN was tested for a number of conditions and compared to those obtained by using deterministic models. Both ANN and deterministic models were validated experimentally. In all cases, at low biomass concentration, model predictions yielded determination coefficients greater than 0.9. Nevertheless, ANN yielded the more accurate predictions of the light pattern, at both low and high biomass concentration, when the bioreactor radius, the depth, the rotational speed of the stirrer and the biomass concentration were incorporated in the ANN structure. In comparison, most of the deterministic models failed to correlate the empirical data at high biomass concentration. These results show the usefulness of ANNs in the modeling of the light profile pattern in photobioreactors.

Tequila industries in general produce great volumes of effluents with high pollutant loads, which are discharged (untreated or partially treated) into natural receivers, thus causing severe environmental problems. In this contribution, we propose an integrated system as a first step to comply with the Mexican ecological norms and stabilize the anaerobic treatment of Tequila vinasses with main design criteria: simple and easy operation, reduce operating time and associated costs (maintenance), integrated and compact design, minimal cost of set-up, start-up, monitoring and control. This system is composed of a fully instrumented and automated lab-scale CSTR-type digester, on-line measuring devices of key variables (pH, temperature, flow rates, etc.), which are used along with off-line readings of chemical oxygen demand (COD), biogas composition, alkalinity and volatile fatty acids to guarantee the operational stability of the anaerobic digestion process. The system performance was evaluated for 200 days and the experimental results show that even under the influence of load disturbances, it is possible to reduce the COD concentration to 85% in the start-up phase and up to 95% during the normal operation phase while producing a biogas with a methane composition greater than 65%. It is also shown that in order to maintain an efficient treatment, the buffering capacity (given by the alkalinity ratio, alpha = intermediate alkalinity/total alkalinity) must be closely monitored.