Applied Microbiology and Biotechnology

, Volume 75, Issue 3, pp 693–701 | Cite as

Quantitative analysis of a high-rate hydrogen-producing microbial community in anaerobic agitated granular sludge bed bioreactors using glucose as substrate

  • Chun-Hsiung HungEmail author
  • Kuo-Shing Lee
  • Lu-Hsiu Cheng
  • Yu-Hsin Huang
  • Ping-Jei Lin
  • Jo-Shu Chang
Environmental Biotechnology


Fermentative H2 production microbial structure in an agitated granular sludge bed bioreactor was analyzed using fluorescence in situ hybridization (FISH) and polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE). This hydrogen-producing system was operated at four different hydraulic retention times (HRTs) of 4, 2, 1, and 0.5 h and with an influent glucose concentration of 20 g chemical oxygen demand/l. According to the PCR-DGGE analysis, bacterial community structures were mainly composed of Clostridium sp. (possibly Clostridium pasteurianum), Klebsiella oxytoca, and Streptococcus sp. Significant increase of Clostridium/total cell ratio (68%) was observed when the reactor was operated under higher influent flow rate. The existence of Streptococcus sp. in the reactor became more important when operated under a short HRT as indicated by the ratio of Streptococcus probe-positive cells to Clostridium probe-positive cells changing from 21% (HRT 4 h) to 38% (HRT 0.5 h). FISH images suggested that Streptococcus cells probably acted as seeds for self-flocculated granule formation. Furthermore, combining the inspections with hydrogen production under different HRTs and their corresponding FISH analysis indicated that K. oxytoca did not directly contribute to H2 production but possibly played a role in consuming O2 to create an anaerobic environment for the hydrogen-producing Clostridium.


Fermentative hydrogen production Granular sludge Bioreactor design Bacterial community structure Fluorescence in situ hybridization 



This work was financially supported by the Bureau of Energy, Taiwan (grant numbers 93-ET-7-006-001-ET and 94-ET-7-006-004-ET).


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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Chun-Hsiung Hung
    • 1
    Email author
  • Kuo-Shing Lee
    • 2
  • Lu-Hsiu Cheng
    • 1
  • Yu-Hsin Huang
    • 3
  • Ping-Jei Lin
    • 3
  • Jo-Shu Chang
    • 4
  1. 1.Department of Environmental EngineeringNational Chung-Hsing UniversityTaichungTaiwan
  2. 2.Department of Safety, Health, and Environmental EngineeringCentral Taiwan University of Science and TechnologyTaichungTaiwan
  3. 3.Department of Chemical EngineeringFeng Chia UniversityTaichungTaiwan
  4. 4.Department of Chemical EngineeringNational Cheng Kung UniversityTainanTaiwan

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