Acclimation of Acid-Tolerant Methanogenic Culture for Bioaugmentation: Strategy Comparison and Microbiome Succession

[Image: see text] To enrich an acid-tolerant methanogenic culture used as bioaugmented seed under acidic conditions, we operated four semicontinuous digesters under various conditions of pH decline for producing methane at pH 5.0. 16S rRNA amplification was performed to unravel the association between declining pH and microbiome succession. The findings demonstrated that a gradual decrease of pH, at a step size of 0.5, and a prolonged run time at each pH could achieve a suitable microbial culture, in which acetoclastic Methanothrix and hydrogenotrophic Methanolinea represented the dominant methanogens. In contrast, a sharp decline in pH could result in heavy loss of the acetoclastic methanogen Methanothrix, leading to a cessation of methane production. Hydrogenotrophic methanogens exhibited high acid tolerance, and Methanospirillum could thrive despite a sudden low-pH shock. Although Methanolinea required a longer time to enrich, it played a substantial role in methane production under an acidic environment.