Biomethane Production and Methanogenic Microbiota Restoration After a pH Failure in an Anaerobic Sequencing Batch Reactor (A-SBR) Treating Tequila Vinasse

Precise control of operational parameters in anaerobic digestion reactors is crucial to avoid imbalances that could affect biomethane production and alterations in the microbiota. Restoring the methanogenic microbiota after a failure is essential for recovering methane production, yet no published strategies exist for this recovery. In this study, we restored the methanogenic microbiota in an anaerobic SBR reactor that operates with both biofilm and suspended biomass simultaneously, aiming to treat tequila vinasses. Four strategies were evaluated for restoring the methanogenic microbiota: reducing the initial vinasse concentration, increasing the reaction time (RT), adjusting the carbon/nitrogen (C/N) ratio, and progressively increasing the initial vinasse concentration. Among these, adjusting the C/N ratio emerged as a critical parameter for restoring organic matter removal efficiency and reestablishing methanogenic microbiota. The operational conditions under which the methanogenic activity and microbiota were restored were as follows: Operating the A-SBR with an initial vinasse concentration of 60%, an RT of 168 h, a pH of 6.9 +/- 0.2, a temperature of 35 +/- 2 degrees C, and a C/N ratio adjusted to 100/1.9 resulted in stable COD removal efficiency of 93 +/- 3% over a year and a high percentage of methanogenic microorganisms in both the suspended microbiota (69%) and biofilm (52%). The normalized methane production (0.332 NL CH4/g CODr) approached the theoretical maximum value (0.35 L CH4/g CODr) after restoring the population and methanogenic activity within the reactor.