Methanogenic consortia from thermophilic molasses-fed structured-bed reactors: microbial characterization and responses to varying food-to-microorganism ratios

The heterogeneous character of fixed-film reactors may create highly specialized zones with a stratified distribution of microbial groups and varying capabilities to withstand high organic loads in anaerobic digestion (AD) systems. The microbial distribution and methane-producing potential of biomass from different regions (feeding zone and structured bed) of two second-stage thermophilic (55 ºC) fixed-film reactors were assessed. Three levels of food-to-microorganism (F/M) ratio (0.4, 1.0 and 3.0 g-COD g(−1)VS) using fermented (two-stage AD) and fresh (single-stage AD) sugarcane molasses were tested in batch reactors, simulating low to high organic loads. Specific methane production rates increased as the F/M increased when using fermented molasses, maintaining efficient methanogenesis at substrate availability levels threefold higher than single-stage schemes (3.0 vs. 1.0 g-COD g(−1)VS). Success in methane production derived from the homogenous establishment (similar in both feeding zone and bed) of syntrophic associations between acetogens (Pelotomaculum, Syntrophothermus, Syntrophomonas and Thermodesulfovibrio), acetate oxidizers (Thermoacetogenium, Mesotoga and Pseudothermotoga) and hydrogenotrophic methogens (Methanothermobacter and Methanoculleus) replacing acetoclastic methanogens (Methanosaeta). Phase separation under thermophilic conditions was demonstrated to boost methane production from sugar-rich substrates, because the process depends on microbial groups (hydrogenotrophs) that grow faster and are less susceptible to low pH values compared to acetotrophs. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s43153-022-00291-x.