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Environmental Microbiology |
Department of Environmental Engineering, National Cheng Kung University, Tainan, Taiwan1
Graduate Institute of Environmental Engineering, National Central University, Chungli 32054, Taiwan2
Department of Biology, National Cheng Kung University, Tainan, Taiwan3
Author for correspondence: Wen-Tso Liu. Tel: +886 3422 7151 ext. 4683. Fax: +886 3426 9401. e-mail: liuwt{at}cc.ncu.edu.tw
The microbial composition and spatial distribution in a terephthalate-degrading anaerobic granular sludge system were characterized using molecular techniques. 16S rDNA clone library and sequence analysis revealed that 78·5% of 106 bacterial clones belonged to the 
subclass of the class Proteobacteria; the remaining clones were assigned to the green non-sulfur bacteria (7·5%), Synergistes (0·9%) and unidentified divisions (13·1%). Most of the bacterial clones in the -Proteobacteria formed a novel group containing no known bacterial isolates. For the domain Archaea, 81·7% and 18·3% of 72 archaeal clones were affiliated with Methanosaeta and Methanospirillum, respectively. Spatial localization of microbial populations inside granules was determined by transmission electron microscopy and fluorescent in situ hybridization with oligonucleotide probes targeting the novel -proteobacterial group, the acetoclastic Methanosaeta, and the hydrogenotrophic Methanospirillum and members of Methanobacteriaceae. The novel group included at least two different populations with identical rod-shape morphology, which made up more than 87% of the total bacterial cells, and were closely associated with methanogenic populations to form a nonlayered granular structure. This novel group was presumed to be the primary bacterial population involved in the terephthalate degradation in the methanogenic granular consortium.
Keywords: denaturing gradient gel electrophoresis, clone library, 16S rDNA, in situ hybridization, UASB
Abbreviations: DGGE, denaturing gradient gel electrophoresis; FISH, fluorescent in situ hybridization; GNS, green non-sulfur; OTU, operational taxonomic unit; TEM, transmission electron microscopy; UASB, upflow anaerobic sludge bed
The GenBank accession numbers for the sequences obtained in this work are AF229774–AF229793.
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