Changes of Microbial Community Depending on Different Dissolved Oxygen in Biological Nitrogen Removal Process

생물학적 질소제거 공정에서 용존산소변화에 따른 미생물의 군집변화

  • Park, Jong-Il (Department of Environmental Engineering, Seoul National University of Technology) ;
  • Lee, Tae-Jin (Department of Environmental Engineering, Seoul National University of Technology)
  • 박종일 (서울산업대학교 환경공학과) ;
  • 이태진 (서울산업대학교 환경공학과)
  • Published : 2008.09.30

Abstract

PCR-DGGE method was applied to analyze changes of microbial community in simultaneous nitrification and denitrification (SND) bioreactor with various DO concentrations. In the analysis of eubacterial community, band profiles of DGGE were similar with 2 or 1 mg/L DO concentrations in the reactor. Experimental results led to 16 different bacteria being identified, including 5 dominant strains(3 strains of Uncultured Bacterium, 1 strains of Bacillus, 1 strains of Uncultured Bacteroidetes). DGGE results at 0.5 mg/L DO concentration led to 12 strains being identified, including 7 dominant strains(5 strains of Uncultured Bacterium, 2 strains of Zoogloea sp.). DGGE results at 0.1 mg/L DO concentration led to 11 strains being identified, including 3 dominant strains(1 strains of Uncultured Bacterium, 2 strains of Zoogloea sp.). In DGGE band profiles of $\beta$-AOB($\beta$-Ammonia Oxidizing Bacteria), only one band was observed. This band had 97% similarity with Nitrosomonas sp. done DNB Y20. This band was clearly observed at the 2, 1 and 0.5 mg/L DO concentrations, while the brightness of the band at 0.1 mg/L DO concentration was mostly dimmed. In DGGE band profiles of denitrification process, 5 bands(3 strains of Uncultured organism containing nirS, 2 strains of Uncultured organism containing nirK) were observed. Among those bands, the brightness of one band was gradually increased at the lower DO concentrations. This band has 86% identity with Uncultured organism clone eS1 cd1 nirS gene, partial cds. Based on this result, it could be concluded that Uncultured organism clone eS1 cd1 nirS gene, partial cds is a predominant microorganism in the denitrification process.

본 연구에서는 PCR-DGGE 기법을 이용하여 DO 농도에 따른 동시 질산화.탈질 반응조 내 미생물 군집 변화 양상을 규명하고자 하였다. DO 농도의 변화에 따른 eubacteria의 군집 변화 해석 실험에서, DO 농도를 2와 1 mg/L로 운전한 반응조 내의 band profile은 거의 유사하게 관찰되었으며 5종의 우점화 미생물(Uncultured Bacterium 3종, Bacillus sp. 1종, Uncultured Bacteroidetes sp. 1종)을 포함한 16종의 미생물을 동정할 수 있었다. 그리고 DO 농도 0.5 mg/L로 운전한 반응조 내의 DGGE 결과 7종의 우점화 미생물(Uncultured Bacterium 5종, Zoogloea sp. 2종)을 포함한 12종의 미생물을 동정할 수 있었으며, DO 농도 0.1 mg/L로 운전한 반응조의 경우 3종의 우점화 미생물(Uncultured Bacterium 1종, Zoogloea sp. 2종)을 포함한 11종의 미생물을 동정할 수 있었다. 반응조 내 DO 농도의 변화에 따른 $\beta$-AOB($\beta$-Ammonia Oxidizing Bacteria)의 군집 변화 해석 실험 결과, 하나의 band를 관찰할 수 있었다. 이 band는 Uncultured Nitrosomonas sp. done DNB Y20와 97%의 유사도를 갖는 것으로 나타났으며 2, 1, 그리고 0.5 mg/L의 DO 농도에서 추출한 sample에서는 선명하게 관측되었으나, 0.1 mg/L DO 농도에서 추출한 sample에서는 선명도가 현저히 감소하였다. 이는 NH$_4{^+}$-N의 질산화 양상과 상관관계가 있음을 보였다. 반응조 내 DO 농도에 따른 탈질 bacteria의 군집 변화 해석실험 결과, 다섯 개의 band(nirS를 함유하는 Uncultured organism 미생물 3종, nirK를 함유하는 Uncultured bacterium 미생물 2종)가 관측되었으며 관측된 band 중 한 band는 DO 농도가 낮아질수록 선명도가 현저히 증가하는 것을 확인할 수 있었다. 이 band에 해당하는 미생물은 86%의 유사도를 가진 Uncultured organism clone eS1 cd1 nirS gene, partial cds로, 본 연구의 탈질 반응에 직접적으로 관여 하는 미생물로 사료된다.

Keywords

References

  1. Metcalf & Eddy, 고광백 외 13인 공역, '폐수처리공학 2," 동화기술, pp. 1048-1248(2004)
  2. Rittman, B. E. and Langeland, W. E., 'Simultaneous Denitrification with Nitrification in Single-Channel Oxidation Ditches,' J. Water pollut. Control Fed., 57, 300(1985)
  3. Trivedi, H. and Heinen, N., 'Simultaneous Nitirification/Denitrification by Mornitoring NAOH Fluorescene in Activated Sludge,' Proceedings of the Facility Operations II: Innovative Technology Forum; 73d Annual Conference, Water Environment Federation, Anaheim, CA(2000)
  4. Irvine, R. L., Murthy, D. V. S., Arora, M. L., Copeman, J. L., Heidman, J. A., 'Analysis of full-scale SBR operation at Grundy Centre, IOWA,' J. Water pollut. Control Fed., 59(3), 132-138(1987)
  5. Masuda, S., Watenabe, Y., Ishiguro, M., 'Biofilm properties and simultaneous nitrification and denitrification in aerobic rotating biological contactors,' Water Sci. Technol., 23, 1355-1363(1991) https://doi.org/10.2166/wst.1991.0588
  6. Halling, S. B., Hjuler, H., 'Simultaneous nitrification and denitrification with an upflow fixed bed reactor applying clinoptililite as media,' Water Treat., 7, 77-88(1992)
  7. Kugleman, I. J., Spector, M., 'Nutrient removal in staged biological reactors,' Proceedings of the third WPCF Joint Technical Seminar on Sewage Treatment Technology, Tokyo, Japan, May 12-14, pp. 4-27(1988)
  8. Moriyama, K., Sato, K., Harada, Y., Washiyama, K., Okamoto, K., 'Simultaneous biological removal of nitrogen and phosphorus using oxic-anaerobic-oxic process,' Water Sci. Technol., 22(7/8), 61-66(1990)
  9. Watanebe, Y., Masuda, S., Ishiguro, M., 'Simultaneous nitrification and denitrification in micro-aerobic biofilms,' Water Sci. Technol., 26(3/4), 511-522(1992) https://doi.org/10.1021/es00027a010
  10. Gupta, S. K., Raja, S. M., Gupta, A. B., 'Simultaneous nitrification and denitrification in a rotating biological contactor,' Environ, Technol., 15, 145-153(1994) https://doi.org/10.1080/09593339409385414
  11. Munch, E. V., Lant, L., Keller, J., 'Simultaneous nitrification and denitrification in bench-scale sequencing batch reactors,' Water Res., 30(2), 277-285(1996) https://doi.org/10.1016/0043-1354(95)00174-3
  12. Sen, P., Dentel, S. K., 'Simultaneous nitrification and denitrification in a fluidized bed reactors,' Water Sci. Technol., 38(1), 247-254(1998)
  13. Menoud, P., Wong, C. H., Robinson, H. A., Farquhar, A., Barford, J. P., Barton, G. W., 'Simultaneous Nitrification and Denitrification Using SIPORAXTMPACKING,' Water Sci. Technol., 40(4/5), 153-160(1999)
  14. Fuerhacker, M., Bauer, H., Ellinger, R., Sree, U., Schmid, H., Zibuschka, F., Puxbaum, H., 'Approach for a novel control strategy for simultaneous nitrification-denitrification in activated sludge reactors,' Water Res., 2499-2506(2000)
  15. Watanebe, T., Hashimoto, S., Kuroda, M., 'Simultaneous nitrification and denitrification in a single reactor using a bioelectro- chemical process,' Water Sci. Technol., 46(4/5), 163-169(2002)
  16. Hibiya, K., Tereda, A., Tsuneda, S., Hirata, A., 'Simultaneous nitrification and denitrification by controlling vertical and horizontal microenvironment in a membrane-aerated biofilm reactor,' J. Biotechnol., 100, 23-32(2003) https://doi.org/10.1016/S0168-1656(02)00227-4
  17. 권문선, 김환홍, '탈질균의 호기성상태에서 탈질 특성에 관한 기초적 연구,' 대한환경공학회지, 143-148(1992)
  18. Holman, J. B. and Wareham, D. G., 'COD, ammonia and dissolved oxygen time profiles in the simultaneous nitrification/denitrification process,' Biochem. Eng. J., 22, 125-133(2005) https://doi.org/10.1016/j.bej.2004.09.001
  19. Pochana, K., Keller, J., 'Study of factors affecting simultaneous nitrification and denitrification(SND),' Water Sci. Technol., 39(6), 61-68(1999)
  20. Pochana, K., Keller, J., Lant, J., 'Model development for simultaneous nitrification and denitrification,' Water Sci. Technol., 39(1), 235-243(1999)
  21. Muyzer, G., Smalla, K., 'Application of denaturing gradient gel electrophoresis(DGGE) and temperature gradient gel electrophoresis(TGGE) in microbial ecology,' Antonie van Leeuwenhoel, 73, 127-141(1998) https://doi.org/10.1023/A:1000669317571
  22. Claudia Schabereiter-Gurtner, Warner Lubitz, Sabine Roleke, 'Application of broad- range 16S rRNA PCR amplication and DGGE fingerprinting for detection of tick- infecting bacteria,' J. Microbiol. Methods, 52, 251-260(2003) https://doi.org/10.1016/S0167-7012(02)00186-0
  23. Emilie Lyautey, Benedicte Lacoste, Loic Ten-Hage, Jean-Luc Rols, Frederic Garabetian, 'Analysis of bacterial diversity in river biofilms using 16S rRNA PCR-DGGE: methodological setting and fingerprints interpretation,' Water Res., 39, 380-388(2005) https://doi.org/10.1016/j.watres.2004.09.025
  24. Muyzer, G., Ellen C., De Waal, Andre G. Uitierlinden, 'Profiling of Complex Microbial Populations by Denaturing Gradient Gel Electrophoresis Analysis of Polymerase Chain Reaction-Amplified Genes Coding for 16S rRNA,' Appl. Environ. Microbiol., 59, 695-700(1993)
  25. Mannix Salvador Pedro, Shin Haruta, Masaru Hazaka, Rumiko Shimada, Chie Yoshida, Koichiro Hiura, Masaharu Ishii, Yasuo Igarashi, 'Denaturing Gradient Gel Electrophoresis Analysis of microbial community from field-scale composter,' J. Bioscience and Bioengineering, 91(2), 159-165(2001) https://doi.org/10.1016/S1389-1723(01)80059-1
  26. 정수희, 'PCR-DGGE를 이용한 호기성 고정생물막내 질산화 미생물의 군집해석,' 부산대학교 대학원 환경공학과 (2005)
  27. 이진옥, 'Real-time PCR을 이용한 BioCube 내부 탈질 미생물 분포 측정,' 대한환경공학회 2005년 추계학술연구발표회지, pp. 164-168(2005)
  28. Gesche Braker, Andreas Fesefeldt, Karl-Paul Witzel, 'Development of PCR primer system for amplication of nitrite reductase genes(nirK and nirS) to detect denitrifying bacteria in environment samples,' Appl. Environ. Microbiol., Oct., 3769-3775(1998)
  29. APWA, AWWA, WPCF, 'Standard methods for the examination of water and wastewater,' 20th ed.(1999)
  30. Christensen, M. H., Harremoes, P., 'Nitrification and denitrification in wastewater treatment,' pp. 391-414, In:Water Pollution Microbiology. 2, R. Mitchell, Ed. John Wiley & Sons, New York(1978)
  31. Garbriel Bitton저, 오계헌, 송홍규, 최철호, 공인철, 정금희, 이병의 공역, '폐수 미생물,' pp. 247-312(2004)