Distribution and abundance of Spirochaetes in full-scale anaerobic digesters
- Distribution and abundance of Spirochaetes in full-scale anaerobic digesters
- 이상훈; 강현진; 이영행; 이택준; 박희등
- Anaerobic digester; FISH; Phylogenetic analysis; Pyrosequencing; Spirochaetes
- Issue Date
- 2012 International Biotechnology Symposium and Exhibition(IBS)
- Spirochaetes are helically coiled Gram-negative bacteria living chemoheterotrophically in anaerobic
condition. Although the bacteria belonging to the Spirochaetes are frequently detected in anaerobic
digesters, the distribution and abundance of them are rarely reported. Especially the phylogenetic position
of the Spirochaetes in anaerobic digesters is not well characterized. In this study, we explored the
distribution and abundance of Spirochaetes in seven full-scale anaerobic digesters located in Seoul,
Korea. Digester sludge samples were collected monthly for six months in year 2010. A total of 83,774
sequences of 16S rRNA gene fragments were retrieved from the samples using the barcode 454
pyrosequencing technology. From the sequences, 9,051 operational taxonomic units (OTUs) were
identified at 3% cutoff, and 494 OTUs were within the Spirochaetes. Spirochaetal OTUs were detected in
all seven anaerobic digesters and the proportion of them was varied from 0.4% to 26% (18% in average).
Spirochaetal OTUs were distributed mainly in anaerobic digesters operated with mesophilic condition
(36-38oC), which was confirmed by redundancy analysis (RDA). RDA evaluated correlations between
the pattern of spirochaetal OTUs and operational factors, and demonstrated that temperature was the only
significant operational factor (P <0.05) affecting the distribution of spirochaetal OTUs. A phylogenetic
analysis revealed that most of spirochaetal OTUs were clustered in phylum Spirochaetes and analyzed as
a novel cluster not reported previously. A fluorescence in situ hybridization (FISH) probe targeting the
spirochaetal cluster was designed to identify and quantify the Spirochaetes in anaerobic digester samples.
The distribution and abundance of Spirochaetes evaluated by FISH experiments were agree with the
results obtained from pyrosequencing, suggesting the Spirochaetes are important players in anaerobic
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