@article{mbs:/content/journal/micro/10.1099/00221287-148-11-3353, author = "Crocetti, Gregory R. and Banfield, Jillian F. and Keller, Jürg and Bond, Philip L. and Blackall, Linda L.", title = "Glycogen-accumulating organisms in laboratory-scale and full-scale wastewater treatment processesbbThe GenBank accession numbers for the sequences reported in this paper are given in Methods.", journal= "Microbiology", year = "2002", volume = "148", number = "11", pages = "3353-3364", doi = "https://doi.org/10.1099/00221287-148-11-3353", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-148-11-3353", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "fluorescence in situ hybridization (FISH)", keywords = "VFA, volatile fatty acid", keywords = "GAOs", keywords = "FISH, fluorescence in situ hybridization", keywords = "EBPR, enhanced biological phosphorus removal", keywords = "PHA, poly-β-hydroxyalkanoate", keywords = "wastewater treatment", keywords = "CLSM, confocal laser scanning microscope/microscopy", keywords = "COD, chemical oxygen demand", keywords = "GAO, glycogen-accumulating organism", keywords = "OTU, operational taxonomic unit", keywords = "PAO, polyphosphate-accumulating organism", keywords = "EBPR", keywords = "SBR, sequencing batch reactor", abstract = "Laboratory-scale sequencing batch reactors (SBRs) as models for wastewater treatment processes were used to identify glycogen-accumulating organisms (GAOs), which are thought to be responsible for the deterioration of enhanced biological phosphorus removal (EBPR). The SBRs (called Q and T), operated under alternating anaerobic–aerobic conditions typical for EBPR, generated mixed microbial communities (sludges) demonstrating the GAO phenotype. Intracellular glycogen and poly-β-hydroxyalkanoate (PHA) transformations typical of efficient EBPR occurred but polyphosphate was not bioaccumulated and the sludges contained 1·8% P (sludge Q) and 1·5% P (sludge T). 16S rDNA clone libraries were prepared from DNA extracted from the Q and T sludges. Clone inserts were grouped into operational taxonomic units (OTUs) by restriction fragment length polymorphism banding profiles. OTU representatives were sequenced and phylogenetically analysed. The Q sludge library comprised four OTUs and all six determined sequences were 99·7% identical, forming a cluster in the γ-Proteobacteria radiation. The T sludge library comprised eight OTUs and the majority of clones were Acidobacteria subphylum 4 (49% of the library) and candidate phylum OP10 (39% of the library). One OTU (two clones, of which one was sequenced) was in the γ-Proteobacteria radiation with 95% sequence identity to the Q sludge clones. Oligonucleotide probes (called GAOQ431 and GAOQ989) were designed from the γ-Proteobacteria clone sequences for use in fluorescence in situ hybridization (FISH); 92% of the Q sludge bacteria and 28% of the T sludge bacteria bound these probes in FISH. FISH and post-FISH chemical staining for PHA were used to determine that bacteria from a novel γ-Proteobacteria cluster were phenotypically GAOs in one laboratory-scale SBR and two full-scale wastewater treatment plants. It is suggested that the GAOs from the novel cluster in the γ-Proteobacteria radiation be named ‘Candidatus Competibacter phosphatis’.", }