Enrichment of thermophilic methanogenic microflora from mesophilic waste activated sludge for anaerobic digestion of garbage slurry

Abstract

This study investigated a startup strategy for thermophilic methanogenic enrichment. Conventional waste activated sludge (WAS) was used as the seed. The WAS seed was incubated at 55 °C in a continuous-flow stirred tank reactor, with garbage slurry fed continuously as a substrate. One of the two reactors (termed reactor-high, RH) was fed with a high concentration of substrate (30 g-COD/L), while the other (reactor-low, RL) received a lower concentration of feed (15 g-COD/L). The specific organic loading rate was 0.2 g-COD/L/day initially, which was gradually increased by shortening the hydraulic retention time. The final OLR was 3.2 g-COD/L/day, after more than 90% of the initial WAS got washed out from the reactor and thermophilic microorganisms became dominant in the reactors. Biogas production rate and methane conversion ratio depended on substrate concentration, although total chemical oxygen demand removal and methane content were almost the same in RH and RL. Biogas production rate in RH was 3.2 times higher than that in RL, while the conversion ratio of RH was 1.6 times higher than that of RL. Quantitative polymerase chain reaction analysis using specific primers for the mcrA gene and high-throughput sequencing analysis of 16S rRNA gene amplicons demonstrated post enrichment differences in the microbial community, relative to that in the WAS. There was no significant difference in the enriched microbial community composition between RH and RL. In conclusion, thermophilic methanogenic microflora can be enriched from mesophilic seeds. Methanothermobacter, Methanosarcina, and other thermophilic bacteria were enriched in the community over time, with these thermophiles collectively accounting for ∼80% of the stable thermophilic community.

Keywords: Anaerobic digestion; Microbial community dynamics; Startup; Thermophilic methanogen; Wastewater treatment.