Laboratory-scale sequencing batch reactors (SBRs) as models for activated sludge processes were used to study enhanced biological phosphorus removal (EBPR) from wastewater. Enrichment for polyphosphateaccumulating organisms (PAOs) was achieved essentially by increasing the phosphorus concentration in the influent to the SBRs. Fluorescence in situ hybridization (FISH) using domain-, division-, and subdivision-level probes was used to assess the proportions of microorganisms in the sludges. The A sludge, a high-performance P-removing sludge containing 15.1% P in the biomass, was comprised of large clusters of polyphosphate-containing coccobacilli. By FISH, >80% of the A sludge bacteria were ␤-2 Proteobacteria arranged in clusters of coccobacilli, strongly suggesting that this group contains a PAO responsible for EBPR. The second dominant group in the A sludge was the Actinobacteria. Clone libraries of PCR-amplified bacterial 16S rRNA genes from three high-performance P-removing sludges were prepared, and clones belonging to the ␤-2 Proteobacteria were fully sequenced. A distinctive group of clones (sharing >98% sequence identity) related to Rhodocyclus spp. (94 to 97% identity) and Propionibacter pelophilus (95 to 96% identity) was identified as the most likely candidate PAOs. Three probes specific for the highly related candidate PAO group were designed from the sequence data. All three probes specifically bound to the morphologically distinctive clusters of PAOs in the A sludge, exactly coinciding with the ␤-2 Proteobacteria probe. Sequential FISH and polyphosphate staining of EBPR sludges clearly demonstrated that PAO probe-binding cells contained polyphosphate. Subsequent PAO probe analyses of a number of sludges with various P removal capacities indicated a strong positive correlation between P removal from the wastewater as determined by sludge P content and number of PAO probe-binding cells. We conclude therefore that an important group of PAOs in EBPR sludges are bacteria closely related to Rhodocyclus and Propionibacter.

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 18 % P (sludge Q) and 15 % P (sludge T). 16S rDNA clone libraries were prepared from DNA extracted from the Q and T sludges.

Host-associated bacterial communities are potentially critical components of marine microbial diversity, yet our understanding of bacterial distribution on living surfaces lags behind that for planktonic communities. We used 16S rRNA gene library analysis to compare within-host (alpha) and between-host (beta) diversity among bacterial communities on 3 co-occurring marine eukaryotes from temperate Australia: the demosponge Cymbastela concentrica, the red macroalga Delisea pulchra and the green intertidal alga Ulva australis. The bacterial community on C. concentrica had high phylum-level diversity (7 phyla including 3 proteobacterial classes) but relatively low 'species' richness (estimated at 24 species). Among the algae, D. pulchra contained 7 phyla including an estimated 79 species, while the U. australis library yielded only 4 phyla with an estimated 36 species. Alpha-, Delta-and Gammaproteobacteria were well represented in all libraries, while Planctomycetes and Bacteroidetes were relatively common on the 2 algae, but absent or rarely encountered in the sponge. At the phylum level, the community of C. concentrica largely mirrored that found in other marine sponges (e.g. Proteobacteria, Actinobacteria, Nitrospira), although large numbers of diatoms and the presence of Verrucomicrobia were atypical. Overall, within-host (alpha) diversity was relatively high, at least for C. concentrica and D. pulchra, while between-host (beta) diversity depended heavily on the phylogenetic level examined. Generally, there was a remarkable lack of overlap at the species level. No species showed universal distribution across hosts, indicating high beta diversity at the species level. At the level of phyla, however, both universal (e.g. Proteobacteria) and distinct (e.g. Nitrospira) groups existed. This study is among the first to compare patterns of alpha and beta diversity for microbial communities associated with co-occurring marine eukaryotes. KEY WORDS: Bacterial communities · Diversity · Marine · Host organisms · Sponge · MacroalgaResale or republication not permitted without written consent of the publisher Aquat Microb Ecol 48: 217-229, 2007 1999, Rappé et al. 2000, Venter et al. 2004, Rusch et al. 2007, with the alphaproteobacterial SAR 11 clade accounting for up to 50% of all microbial cells in ocean surface waters (Morris et al. 2002). Cyanobacteria of the genera Synechococcus and Prochlorococcus are also extremely prevalent (Partensky et al. 1999, Scanlan & West 2002, while other frequently encountered marine taxa include the Bacteroidetes, Actinobacteria, Planctomycetes and Chloroflexi (Giovannoni et al. 1996, Simon et al. 1999, Cottrell & Kirchman 2000, Venter et al. 2004, Giovannoni & Stingl 2005, Rusch et al. 2007). This apparent global distribution of various bacterial phyla (and even particular species, e.g. Mullins et al. 1995 suggests a reasonable degree of uniformity to marine bacterioplankton communities. Despite this overall uniformity, however, compositional differences have also been observe...

Marine Ulvacean algae are colonized by dense microbial communities predicted to have an important role in the development, defense and metabolic activities of the plant. Here we assess the diversity and seasonal dynamics of the bacterial community of the model alga Ulva australis to identify key groups within this epiphytic community. A total of 48 algal samples of U. australis that were collected as 12 individuals at 3 monthly intervals, were processed by applying denaturing gradient gel electrophoresis (DGGE), and three samples from each season were subjected to catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH). CARD-FISH revealed that the epiphytic microbial community was comprised mainly of bacterial cells (90%) and was dominated by the groups Alphaproteobacteria (70%) and Bacteroidetes (13%). A large portion (47%) of sequences from the Alphaproteobacteria fall within the Roseobacter clade throughout the different seasons, and an average relative proportion of 19% was observed using CARD-FISH. DGGE based spatial (between tidal pools) and temporal (between season) comparisons of bacterial community composition demonstrated that variation occurs. Between individuals from both the same and different tidal pools, the variation was highest during winter (30%) and between seasons a 40% variation was observed. The community also includes a sub-population of bacteria that is consistently present. Sequences from excised DGGE bands indicate that members of the Alphaproteobacteria and the Bacteroidetes are part of this stable sub-population, and are likely to have an important role in the function of this marine epiphytic microbial community.

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