Candidatus Monilibacter spp., common bulking filaments in activated sludge, are members of Cluster III Defluviicoccus.

Two alphaproteobacterial Neisser negative 'Nostocoida limicola' morphotypes differing slightly in their trichome diameter and filament regularity were dominant populations in the Bendigo, Victoria, Australia activated sludge community removing phosphorus (P). Neither responded to the FISH probes available for any of the other alphaproteobacterial 'N. limicola' morphotypes. Instead both fluoresced with the DF988 FISH probe designed originally to target alphaproteobacterial cluster II Defluviicoccus tetrad forming organisms. A 16S rRNA based clone library from this biomass revealed that the alphaproteobacterial clones grouped closely with Candidatus 'Monilibacter batavus' and Defluviicoccus clones in a cluster separate from the existing cluster I and II Defluviicoccus. When a FISH probe was designed against these, it only hybridized to the thinner and less abundant 'N. limicola' morphotype. Micromanipulation-RT-PCR was used to selectively recover the main 'N. limicola' morphotype and a FISH probe designed against the 16S rRNA clones generated from it showed only this filament fluoresced. From FISH based surveys, both 'N. limicola' variants occurred frequently in phosphorus removal activated sludge systems in Australia treating domestic waste. The data suggest that they represent two new strains of Candidatus 'Monilibacter', which on this evidence are filamentous members of the genus Defluviicoccus, a potential competitor for the polyphosphate accumulating organisms in these communities.

Nittami T ，McIlroy S ，Seviour EM ，Schroeder S ，Seviour RJ ... -  《-》

Filamentous members of cluster III Defluviicoccus have the in situ phenotype expected of a glycogen-accumulating organism in activated sludge.

The in situ ecophysiology of alphaproteobacterial filamentous Cluster III Defluviicoccus present in enhanced biological phosphorus removal (EBPR)-activated sludge systems was evaluated using FISH-MAR and histochemical staining methods. These organisms, sharing the Nostocoida limicola morphotype, are known to be responsible for serious episodes of activated sludge bulking. The data presented here also demonstrate an ability to assimilate short-chain fatty acids and synthesize poly-β-hydroxyalkanoates (PHA) anaerobically, and then utilize this stored PHA under aerobic conditions, but with no corresponding synthesis of polyphosphate. These features are consistent with an in situ phenotype of glycogen-accumulating organisms (GAO), populations thought to lower the efficiency of EBPR systems by outcompeting polyphosphate-accumulating organisms (PAO) for substrates in their anaerobic feed phase. Survey data indicate that these GAO are as commonly seen as the known PAO in full-scale EBPR-activated sludge systems, which suggest that they might play important roles there, and therefore should not be viewed just as laboratory curiosities.

McIlroy SJ ，Nittami T ，Seviour EM ，Seviour RJ ... -  《-》

FACS enrichment and identification of floc-associated alphaproteobacterial tetrad-forming organisms in an activated sludge community.

McIlroy S ，Hoefel D ，Schroeder S ，Ahn J ，Tillett D ，Saint C ，Seviour RJ ... -  《FEMS MICROBIOLOGY LETTERS》

Phylogeny of the filamentous bacterium 'Nostocoida limicola' III from activated sludge.

:Five strains of the filamentous bacterium 'Nostocoida limicola' III were successfully isolated into pure culture from samples of activated sludge biomass from five plants in Australia. 16S rRNA gene sequence analyses showed that all isolates were members of the Planctomycetales, most closely related to Isosphaera pallida, but they differed phenotypically from this species in that they did not glide and were not thermotolerant. The ultrastructure of these 'N. limicola' III isolates was also consistent with them being Planctomycetales, in that they possessed complex intracellular membrane systems compartmentalizing the cells. However, the arrangements of these intracellular membranes differed between isolates. These data confirm that 'N. limicola' III is phylogenetically unrelated to both 'N. limicola' I and 'N. limicola' II, activated sludge filamentous bacteria which share morphological features in common with 'N. limicola' III and which have been presumed historically to be the same or very similar bacteria.

Liu JR ，McKenzie CA ，Seviour EM ，Webb RI ，Blackall LL ，Saint CP ，Seviour RJ ... -  《international journal of systematic and evolutionary microbiology》

The "Nostocoida limicola" story: resolving the phylogeny of this morphotype responsible for bulking in activated sludge.

On the basis of 16S rRNA sequence analyses of several isolates of "Nostocoida limicola" from activated sludge plants in Australia and other countries, it is clear that "N. limicola" I, II and III are not three morphological variants of a single bacterium but at least three phylogenetically different bacteria. Data show that "N. limicola" I are members of at least two genera in the low mol% G+C gram-positive bacteria, while some isolates of "N. limicola" II belong to the high mol% G+C gram positive bacteria, and "N. limicola" III is a member of the Planctomycetales. Design and application of 16S rRNA targeted probes for each to biomass samples suggests that their phylogeny is more diverse than pure culture studies would suggest.

Seviour RJ ，Liu JR ，Seviour EM ，McKenzie CA ，Blackall LL ，Saint CP ... -  《WATER SCIENCE AND TECHNOLOGY》