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Applied and Environmental Microbiology, July 2006, p. 4942-4949, Vol. 72, No. 7
0099-2240/06/$08.00+0     doi:10.1128/AEM.02985-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Phenotypic Properties and Microbial Diversity of Methanogenic Granules from a Full-Scale Upflow Anaerobic Sludge Bed Reactor Treating Brewery Wastewater{dagger}

Emiliano E. Díaz,1 Alfons J. M. Stams,2 Ricardo Amils,1 and José L. Sanz1*

Department of Molecular Biology, Autonoma University of Madrid, 28049 Madrid, Spain,1 Laboratory of Microbiology, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands2

Received 19 December 2005/ Accepted 20 April 2006

Methanogenic granules from an anaerobic bioreactor that treated wastewater of a beer brewery consisted of different morphological types of granules. In this study, the microbial compositions of the different granules were analyzed by molecular microbiological techniques: cloning, denaturing gradient gel electrophoresis and fluorescent in situ hybridization (FISH), and scanning and transmission electron microscopy. We propose here that the different types of granules reflect the different stages in the life cycle of granules. Young granules were small, black, and compact and harbored active cells. Gray granules were the most abundant granules. These granules have a multilayer structure with channels and void areas. The core was composed of dead or starving cells with low activity. The brown granules, which were the largest granules, showed a loose and amorphous structure with big channels that resulted in fractured zones and corresponded to the older granules. Firmicutes (as determined by FISH) and Nitrospira and Deferribacteres (as determined by cloning and sequencing) were the predominant Bacteria. Remarkably, Firmicutes could not be detected in the brown granules. The methanogenic Archaea identified were Methanosaeta concilii (70 to 90% by FISH and cloning), Methanosarcina mazei, and Methanospirillum spp. The phenotypic appearance of the granules reflected the physiological condition of the granules. This may be valuable to easily select appropriate seed sludges to start up other reactors.

* Corresponding author. Mailing address: Universidad Autónoma de Madrid, Campus de Cantoblanco, Centro de Biología Molecular, E. Biológicas C-101, 28049 Madrid, España. Phone: 34-914.978.078. Fax: 34-914.978.087. E-mail: joseluis.sanz{at}uam.es.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, July 2006, p. 4942-4949, Vol. 72, No. 7
0099-2240/06/$08.00+0     doi:10.1128/AEM.02985-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Satoh, H., Miura, Y., Tsushima, I., Okabe, S. (2007). Layered Structure of Bacterial and Archaeal Communities and Their In Situ Activities in Anaerobic Granules. Appl. Environ. Microbiol. 73: 7300-7307 [Abstract] [Full Text]  


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