Distribution and abundance of Spirochaetes in full-scale anaerobic digesters

Distribution and abundance of Spirochaetes in full-scale anaerobic digesters
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 digestion.
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