Towards exposure of elusive metabolic mixed-culture processes: the application of metaproteomic analyses to activated sludge

2006 ◽  
Vol 54 (1) ◽  
pp. 217-226 ◽  
Author(s):  
P. Wilmes ◽  
P.L. Bond
Keyword(s):  
Protein Expression ◽  
Activated Sludge ◽  
Phosphorus Removal ◽  
Batch Reactor ◽  
Biological Phosphorus Removal ◽  
Microbial Community Structures ◽  
Proteomic Investigation ◽  
Two Phases ◽  
Biological Phosphorus

Protein expression is a direct reflection of specific microbial activities in any ecosystem. In order to assess protein expression in mixed microbial communities, the feasibility of applying proteomic techniques to activated sludge samples has recently been demonstrated. We report the application of metaproteomics to two activated sludges from a laboratory-scale sequencing batch reactor with dissimilar phosphorus removal performances. Fluorescence in situ hybridization (FISH) revealed that the sludge with good enhanced biological phosphorus removal performance (EBPR) was dominated by Betaproteobacteria (65% of EUBMIX binding cells) and gave positive signals for the Rhodocyclus-type PAO specific probe (59%). The non-EBPR sludge was dominated by tetrad-forming Alphaproteobacteria (75%). With regard to the proteomic investigation, 630 individual protein spots were matched across the replicate groups of the anaerobic and aerobic phases of the EBPR sludge with 9.4% of all spots being statistically different between the two phases. The non-EBPR metaproteomic maps exhibited 590 matched spots with 14.7% statistical differences between the two phases. Overall, the non-EBPR sludge expressed around 30% more significant differences than the EBPR sludge. The comparison of protein expression in the two sludges showed that their metaproteomes were substantially different and this was reflected in their microbial community structures and metabolic transformations.

Morphology, In-Situ characterization with rRNA targetted probes and respiratory quinone profiles of enhanced biological phosphorus removal sludge

1998 ◽  
Vol 38 (8-9) ◽  
pp. 69-76 ◽  
Author(s):  
I. M. Sudiana ◽  
T. Mino ◽  
H. Satoh ◽  
T. Matsuo
Keyword(s):  
Carbon Source ◽  
Activated Sludge ◽  
Microbial Communities ◽  
Phosphorus Removal ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
In Situ Characterization ◽  
Respiratory Quinone ◽  
Biological Phosphorus

The microbial communities in activated sludge acclimated with either acetate or glucose as the major carbon source under phosphorus limited or rich conditions were investigated morphologically, phylogenetically and chemotaxonomically. The sludge with a minimized polyphosphate content was dominated by tetrad shaped bacteria, which were suspected to be ‘glycogen accumulating bacteria (GAOs) or G bacteria’ The sludge containing high polyphosphate was dominated by cluster forming coccus bacteria. Quinone analyses suggested that all the sludge tested contained various ubiquinones and menaquinones, of which the ubiquinones Q-8 and Q-10 were dominant. Analyses with rRNA targeted probes showed that beta sub class of Proteobacteria was most predominant in all sludges tested. Morphological, phylogenetic and chemotaxonomic investigation all indicated that both high and low P sludges are microbiologically diverse.

pH: Keyfactor in the Biological Phosphorus Removal Process

1994 ◽  
Vol 29 (7) ◽  
pp. 71-74 ◽  
Author(s):  
G. J. F. Smolders ◽  
M. C. M. van Loosdrecht ◽  
J. J. Heijnen
Keyword(s):  
Activated Sludge ◽  
Phosphorus Removal ◽  
Sequencing Batch Reactor ◽  
Batch Reactor ◽  
Glycogen Metabolism ◽  
Activated Sludge Process ◽  
Biological Phosphorus Removal ◽  
Anaerobic Conditions ◽  
Biochemical Mechanisms ◽  
Biological Phosphorus

Experiments have been performed, using a sequencing batch reactor, to examine the effect of pH on biological phosphorus removal in the activated sludge process. The results, which indicate that glycogen metabolism occurs during anaerobic conditions, are useful in elucidating the biochemical mechanisms involved in phosphorus-removal, and have potential implications for systems such as Phostrip.

The bioaugmentation of sequencing batch reactor sludges for biological phosphorus removal

1997 ◽  
Vol 35 (1) ◽  
pp. 19-26 ◽  
Author(s):  
E. Belia ◽  
P. G. Smith
Keyword(s):  
Activated Sludge ◽  
Nitrogen Removal ◽  
Pure Culture ◽  
Phosphorus Removal ◽  
Sequencing Batch Reactor ◽  
Batch Reactor ◽  
Sewage Treatment Plant ◽  
Biological Phosphorus Removal ◽  
Conventional Activated Sludge ◽  
Biological Phosphorus

The development of enhanced biological phosphorus removal (EBPR) through the bioaugmentation of a conventional activated sludge was studied. The objectives of the study were to evaluate the phosphorus removal capability of a sequencing batch reactor (SBR) when started with conventional activated sludge and augmented with a pure culture of Acinetobacter lwoffii. The effect of the addition of the pure culture on the reactor start up time, the settling properties of the sludge and on COD and nitrogen removal was also investigated. The effect of the removal of up to 70% of the bioaugmented biomass and its substitution with unconditioned sludge from a conventional sewage treatment plant was determined. This study has demonstrated that bioaugmentation can convert a conventional sewage works activated sludge to an EBPR sludge in 14 days. The sludge produced shows resilience to influent phosphate fluctuations, low D.O. and biomass replacement. The COD and nitrogen removal capabilities of the sludge and its settling properties are not affected by the addition of the pure culture.

Analysis of microbial community that performs enhanced biological phosphorus removal in activated sludge fed with acetate

2002 ◽  
Vol 46 (1-2) ◽  
pp. 145-154 ◽  
Author(s):  
M. Onuki ◽  
H. Satoh ◽  
T. Mino
Keyword(s):  
Microbial Community ◽  
Activated Sludge ◽  
Phosphorus Removal ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Batch Reactor ◽  
Phosphate Removal ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Dominant Band ◽  
Biological Phosphorus

Enhanced biological phosphorus removal (EBPR) activated sludge was operated in a laboratory-scale sequencing batch reactor (SBR) fed with acetate as the sole carbon source. The microbial community of the sludge was analyzed using the polymerase chain reaction (PCR) – denaturing gradient gel electrophoresis (DGGE) method for about 2 months of start-up period. As a result, the number of major bands decreased during the enrichment, indicating that the microbial community structure was getting simpler. Since the phosphate removal activity was maintained at a high level, the bacteria which still remained at the end can be considered as the important bacteria playing key roles in the present EBPR sludge, maybe polyphosphate accumulating organisms (PAOs). The dominant band in the last sample on the DGGE gel was excised and the DNA recovered from it was sequenced. The sequence was closely related to one of the putative PAOs group which Crocetti et al. (2000) and Hesselmann et al. (1999) have proposed. This PAOs group is closely related to the Rhodocyclus group (b-Proteobacteria). The fluorescence in situ hybridization (FISH) method with the probe specific for this PAOs group and the DAPI staining at a phosphate-probing concentration indirectly showed that these Rhodocyclus related bacteria really accumulated polyphosphate.

scholarly journals Identification of a novel subgroup of uncultured gammaproteobacterial glycogen-accumulating organisms in enhanced biological phosphorus removal sludge

Microbiology ◽  
2011 ◽  
Vol 157 (8) ◽  
pp. 2287-2296 ◽  
Author(s):  
Jeong Myeong Kim ◽  
Hyo Jung Lee ◽  
Dae Sung Lee ◽  
Kangseok Lee ◽  
Che Ok Jeon
Keyword(s):  
Phosphorus Removal ◽  
Batch Reactor ◽  
Full Scale ◽  
Rrna Gene ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Phenotypic Properties ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

The presence of glycogen-accumulating organisms (GAO) has been hypothesized to be a cause of deterioration in enhanced biological phosphorus removal (EBPR) processes due to their abilities to out-compete polyphosphate-accumulating organisms (PAO). Based on 16S rRNA gene sequences, new members of uncultured gammaproteobacterial GAO (GB) were identified from sludge samples of a lab-scale sequencing batch reactor used for EBPR. The new GB formed a phylogenetic lineage (GB8) clearly distinct from the previously reported seven GB subgroups. Because the new GB8 members were not targeted by the known fluorescence in situ hybridization (FISH) oligonucleotide probes, a GB8-specific FISH probe (GB429) and a new FISH probe (GB742) targeting all eight GB subgroups were designed, and the phenotypic properties of the new GB8 members were investigated. FISH and microautoradiography approaches showed that GB429-targeted cells (GB8) were large coccobacilli (2–4 µm in size) with the ability to take up acetate under anaerobic conditions, but unable to accumulate polyphosphate under the subsequent aerobic conditions, consistent with in situ phenotypes of GB. FISH analyses on several sludge samples showed that members of GB8 were commonly detected as the majority of GB in lab- and full-scale EBPR processes. In conclusion, this study showed that members of GB8 could be a subgroup of GB with an important role in EBPR deterioration. Designs of FISH probes which hybridize with broader GB subgroups at different hierarchical levels will contribute to studies of the distributions and ecophysiologies of GB in lab- or full-scale EBPR plants.

Comparison of nutrient-removing microbial communities in activated sludge from full-scale MBRs and conventional plants

2013 ◽  
Vol 68 (2) ◽  
pp. 366-371 ◽  
Author(s):  
A. M. Saunders ◽  
P. Larsen ◽  
P. H. Nielsen
Keyword(s):  
Activated Sludge ◽  
Microbial Communities ◽  
Phosphorus Removal ◽  
Membrane Bioreactors ◽  
Full Scale ◽  
Biological Phosphorus Removal ◽  
Conventional Activated Sludge ◽  
16S Rrna Pyrosequencing ◽  
Biological Phosphorus

The composition of nutrient-removing microbial communities in five full-scale membrane bioreactors (MBRs) was investigated using fluorescence in situ hybridization and 16S rRNA pyrosequencing and compared to similar analyses of conventional activated sludge (CAS) communities. The communities were highly similar but some genera that are always present in enhanced biological phosphorus removal (EBPR) (core groups) were absent in the MBRs. The overall phylogenetic similarity of the communities indicated that these differences were primarily closely related groups. More research is needed to establish the operational significance of the observed differences between MBR and CAS sludge.

Population dynamics in wastewater treatment plants with enhanced biological phosphorus removal operated with and without nitrogen removal

2002 ◽  
Vol 46 (1-2) ◽  
pp. 163-170 ◽  
Author(s):  
N. Lee ◽  
J. la Cour Jansen ◽  
H. Aspegren ◽  
M. Henze ◽  
P.H. Nielsen ◽  
...  
Keyword(s):  
Population Dynamics ◽  
Activated Sludge ◽  
Nitrogen Removal ◽  
Phosphorus Removal ◽  
The Other ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Biological Phosphorus ◽  
P Removal

The population dynamics of activated sludge in a pilot plant with two activated sludge systems, both designed for enhanced biological phosphorus removal (EBPR), but one of them with (BNP) and the other without (BP) nitrogen removal, was monitored during a period of 2.5 years. The influent water to the pilot plant was periodically manipulated by external addition of phosphorus (P), acetate and glucose, respectively. The population dynamics and the in situ physiology were monitored by quantitative fluorescence in situ hybridization (FISH) and microautoradiography. Significant P removal was observed in both systems throughout the whole period, with significant increases of the P removal when substrates were dosed. The activated sludge in both systems contained large amounts of dense clusters of gram-negative, methylene-blue staining coccoid rods during the whole period. A large part of the clusters belonged to the β Proteobacteria, whereas the rest of the clusters belonged either to the Actinobacteria or to the α Proteobacteria. The relative abundance of Rhodocyclus-related bacteria in the activated sludge varied significantly in both systems during the whole period (from 6 to 18% in BNP, and from 4 to 28% in BP). However, no statistically significant correlation of the Rhodocyclus-related nor any of the other investigated bacterial groups to the P content of the activated sludge (correlation for all groups investigated was always < 0.5) was observed. A significant 33Pi uptake was observed by the β Proteobacteria (part of them Rhodocyclus-related, the identity of the rest unknown) and the Actinobacteria. However, not all of the Rhodocyclus-related bacteria showed 33Pi uptake. The P removal in the investigated plants is thus believed to be mediated by a mixed population consisting of a part of the Rhodocyclus-related bacteria, the Actinobacteria and other, yet unidentified bacteria.

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