Qualitative and Quantitative Analysis of Microbial Community Structure in the Sequencing Batch Reactor for Enriching ANAMMOX Consortium

연속회분식 반응기를 이용한 혐기성 암모늄 산화균 농후배양에서의 정성 및 정량적 미생물 군집구조 분석

  • Bae, Hyo-Kwan (Center for Environmental Technology Research, Korea Institute of Science and Technology) ;
  • Jung, Jin-Young (Center for Environmental Technology Research, Korea Institute of Science and Technology)
  • 배효관 (한국과학기술연구원 환경기술연구단) ;
  • 정진영 (한국과학기술연구원 환경기술연구단)
  • Received : 2009.07.21
  • Accepted : 2009.09.11
  • Published : 2009.10.31

Abstract

Enrichment of anaerobic ammonium oxidation (ANAMMOX) bacteria is the essential step for operating full-scale ANAMMOX bioreactor because adding a significant amount of seeding sludge is required to stabilize the ANAMMOX reactor. In this study, the enrichment of ANAMMOX bacteria from an activated sludge using sequencing batch reactor was conducted and verified by analyzing changes in the microbial community structure. ANAMMOX bacteria were successfully enriched for 70 days and the substrate removal efficiencies showed 98.5% and 90.7% for $NH_4\;^+$ and $NO_2\;^-$ in the activity test, respectively. The phylogenetic trees of Planctomycetes phylum showed that the diverse microbial community structure of an activated sludge was remarkably simplified after the enrichment. All 36 clones, obtained after the enrichment, were affiliated with ANAMMOX bacteria of Candidatus Brocadia (36%) and Candidatus Anammoxoglobus (64%) genera. The quantification using real-time quantitative PCR (RTQ-PCR) revea ed that the 16S rDNA concentration of ANAMMOX bacteria was 74.8% compared to the granular ANAMMOX sludge obtained from an upflow ANAMMOX sludge bed reactor which had been operated for more than one year. The results of molecular analysis supported that the enriched sludge could be used as a seeding sludge for a full-scale ANAMMOX bioreactor.

혐기성 암모늄 산화공정을 안정화시키기 전에 많은 양의 식종 미생물 투여가 필요하므로 혐기성 암모늄 산화균의 농후배양은 실규모의 혐기성 암모늄 산화 반응기를 운영할 때 필수적인 과정이다. 본 연구에서는 활성슬러지 미생물을 식종한 연속 회분식 반응기를 이용하여 혐기성 암모늄 산화균을 농후배양하고, 미생물 군집구조의 변화를 관찰하여 농후배양 결과를 검증하였다. 혐기성 암모늄 산화균의 농후배양은 70일간 시행되었고, 농후배양 후 활성시험에서 $NH_4\;^+$$NO_2\;^-$의 기질제거효율이 각각 98.5%와 90.7%로 관찰되어 혐기성 암모늄 산화균의 배양이 성공적으로 수행된 것으로 판단되었다. 계통분류학적 분지도 작성 결과, 다양하였던 Planctomycetes 문(phylum)의 미생물 군집구조가 농후배양 이후에 현저하게 단순해졌다는 것이 밝혀졌다. 농후배양 이후 발견된 36개의 clone들 모두가 혐기성 암모늄 산화균이었으며, Candidatus Brocadia (36%) 와 Candidatus Anammoxoglobus (64%) 속(genus)에 속하였다. RTQ-PCR (real-time quantitative PCR)을 통해 혐기성 암모늄 산화균을 정량한 결과, 혐기성 암모늄 산화 상향류식 연속 배양기에서 1년 이상 선택 배양된 붉은색 혐기성암모늄 산화 입상 슬러지에 비해 혐기성 암모늄 산화균의 16S rDNA 농도가 74.8%인 것으로 나타났다. 상기의 분자생물학적 분석을 통해 70일간 농후배양된 활성슬러지가 혐기성 암모늄 산화 실용화 공정의 접종 미생물로 활용 가능할 것으로 판단되었다.

Keywords

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