Determination of the archaeal and bacterial communities in two-phase and single-stage anaerobic systems by 454 pyrosequencing

Yogananda Maspolim, Yan Zhou, Chenghong Guo*, Keke Xiao, Wun Jern Ng

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

36 Scopus citations


2-Phase anaerobic digestion (AD), where the acidogenic phase was operated at 2. day hydraulic retention time (HRT) and the methanogenic phase at 10. days HRT, had been evaluated to determine if it could provide higher organic reduction and methane production than the conventional single-stage AD (also operated at 12. days HRT). 454 pyrosequencing was performed to determine and compare the microbial communities. The acidogenic reactor of the 2-phase system yielded a unique bacterial community of the lowest richness and diversity, while bacterial profiles of the methanogenic reactor closely followed the single-stage reactor. All reactors were predominated by hydrogenotrophic methanogens, mainly Methanolinea. Unusually, the acidogenic reactor contributed up to 24% of total methane production in the 2-phase system. This could be explained by the presence of Methanosarcina and Methanobrevibacter, and their activities could also help regulate reactor alkalinity during high loading conditions through carbon dioxide production. The enrichment of hydrolytic and acidogenic Porphyromonadaceae, Prevotellaceae, Ruminococcaceae and unclassified Bacteroidetes in the acidogenic reactor would have contributed to the improved sludge volatile solids degradation, and ultimately the overall 2-phase system's performance. Syntrophic acetogenic microorganisms were absent in the acidogenic reactor but present in the downstream methanogenic reactor, indicating the retention of various metabolic pathways also found in a single-stage system. The determination of key microorganisms further expands our understanding of the complex biological functions in AD process.

Original languageEnglish
Pages (from-to)121-129
Number of pages9
JournalJournal of Environmental Sciences
StatePublished - 1 Oct 2015
Externally publishedYes


  • 2-Phase anaerobic digestion
  • 454 pyrosequencing
  • Methanogen
  • Microbial community
  • Sewage sludge


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