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Microbiology 150 (2004), 3741-3748; DOI  10.1099/mic.0.27291-0
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Microbiology 150 (2004), 3741-3748; DOI  10.1099/mic.0.27291-0
© 2004 Society for General Microbiology

Identification and occurrence of tetrad-forming Alphaproteobacteria in anaerobic–aerobic activated sludge processes

Man-Tak Wong, Fea Mein Tan, Wun Jern Ng and Wen-Tso Liu

Environmental Science and Engineering Program and Department of Civil Engineering, National University of Singapore, Singapore 117576

Correspondence
Wen-Tso Liu
cveliuwt{at}nus.edu.sg

In an acetate-fed anaerobic–aerobic membrane bioreactor, a deteriorated enhanced biological phosphorus removal (EBPR) community was developed (as determined based on the chemical profiles of organic substrate, soluble phosphate, and intracellular carbohydrate and polyhydroxyalkanote (PHA) concentrations). Microscopic observations revealed the dominance of tetrad-forming organisms (TFOs), of which the majority stained positively for PHA under anaerobic conditions. Fluorescence in situ hybridization (FISH) confirmed that the Alphaproteobacteria (85·0±7·0 % of total cells) were the most dominant group. A 16S rRNA gene clone library specific for the Alphaproteobacteria indicated that most 16S rRNA gene clones (61 % of total clones) were closely affiliated with ‘Defluvicoccus vanus’, forming a cluster within subgroup 1 of the Alphaproteobacteria. Combined PHA staining and FISH with specific probes designed for the members of the ‘Defluvicoccus’ cluster suggested diversity within this TFO cluster, and that these TFOs were newly identified glycogen-accumulating organisms in EBPR systems. However, these ‘Defluvicoccus’-related TFOs were only seen in low abundance in 12 different EBPR and non-EBPR systems, suggesting that they were not the key populations responsible for the deterioration of full-scale EBPR processes.

Abbreviations: CLSM, confocal laser scanning microscopy; DAPI, 4',6-diamidino-2-phenylindole; EBPR, enhanced biological phosphorus removal; FISH, fluorescence in situ hybridization; GAO, glycogen-accumulating organism; HRT, hydraulic retention time; MBR, membrane bioreactor; MLSS, mixed liquor suspended solids; OTU, operational taxonomic unit; PAO, polyphosphate-accumulating organism; PHA, polyhydroxyalkanote; Pi, orthophosphate; SRT, sludge retention time; TOC, total organic carbon; TFO, tetrad-forming organism; VSS, volatile suspended solid

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences reported in this paper are AY351635–AY351641 and AY351643.




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