Density separation and molecular methods to characterize enhanced biological phosphorus removal system populations

2002 ◽  
Vol 46 (1-2) ◽  
pp. 195-198 ◽  
Author(s):  
A.J. Schuler ◽  
M. Onuki ◽  
H. Satoh ◽  
T. Mino
Keyword(s):  
Phosphorus Removal ◽  
Denaturing Gradient Gel Electrophoresis ◽  
High Density ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
High Concentration ◽  
Density Fractions ◽  
Density Separation ◽  
Storage Products ◽  
Biological Phosphorus

A novel approach to the identification of microorganisms that accumulate high density microbial storage products based on density separation, denaturing gradient gel electrophoresis (DGGE), and DNA sequencing was developed and applied to bench and pilot scale enhanced biological phosphorus removal (EBPR) systems. Polyphosphate (PP), glycogen, and polyhydroxyalkanoates (PHAs), are all of higher density than a typical bacterial cell. PP-accumulating organisms (PAOs), the organisms responsible for EBPR, accumulate all three of these storage products. Density separation in a homogenous solution of Percoll produced a high-density biomass fraction with a relatively high concentration of PAOs, as determined by Neisser staining. DNA was extracted from these fractions, amplified, and separated by DGGE. DGGE profiles demonstrated some bacterial strains were present at a greater concentration in the high density fractions than in low density fractions. These strains were considered PAO candidates. 5 of 12 PAO candidates from high density fractions were γ Proteobacteria and only 1 was a β Proteobacterium. 2 PAO candidates were most similar to recently identified γ Proteobacteria sequences obtained by DGGE analysis of a deteriorated benchtop EBPR system.

scholarly journals Performance of enhanced biological phosphorus removal and population dynamics of phosphorus accumulating organisms in sludge-shifting sequencing batch reactors

2018 ◽  
Vol 78 (4) ◽  
pp. 886-895 ◽  
Author(s):  
Yang Pan ◽  
Wenquan Ruan ◽  
Yong Huang ◽  
Qianqian Chen ◽  
Hengfeng Miao ◽  
...  
Keyword(s):  
Removal Efficiency ◽  
Phosphorus Removal ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Batch Reactor ◽  
Batch Reactors ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
High Concentration ◽  
Gradient Gel Electrophoresis ◽  
Biological Phosphorus

Abstract The sludge-shifting sequencing batch reactor (SBR) is an enhanced biological phosphorus removal (EBPR) process for wastewater treatment. In this study, the enrichment of phosphorus accumulating organisms (PAOs) will be attempted by using different high concentration of substrates. In sludge-shifting SBR, activated sludge can be continuously shifted from the bottom of SBR to anaerobic zone/selector, which contains high concentration of substrates, through an orderly reflux between the paralleled SBRs. Denaturing gradient gel electrophoresis (DGGE) methods were used to monitor microbial diversity in sludge. Fluorescence in situ hybridization (FISH) was used to determine the microbial population profile and distribution map under different sludge shifting volumes. The synthesis of intracellular polymers in this process was also analyzed. Phosphorus removal efficiency as high as 96% ± 1.3% was achieved under a sludge shifting ratio of 30%. Synthetic efficiencies of polyhydroxybutyrate (PHB) by PAOs were improved at high sludge shifting ratios. FISH results demonstrated that the population of PAOs in the process increased under properly sludge shifting ratio and it significantly improved phosphorus removal efficiency. Sequencing results indicated that determined sequences (11 OTUs) belonged to Proteobacterium, Actinobacteria and Firmicutes, Pseudomonas kuykendallii, which played an important role in the process of P removal.

Distributed microbial state effects on competitionin enhanced biological phosphorus removal systems

2006 ◽  
Vol 54 (1) ◽  
pp. 199-207 ◽  
Author(s):  
A.J. Schuler
Keyword(s):  
Phosphorus Removal ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Storage Product ◽  
Distributed Approach ◽  
Average System ◽  
Storage Products ◽  
Profile Characteristics ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

Computer simulation of activated sludge population dynamics is a useful tool in process design, operation, and troubleshooting, but currently available programs rely on the assumption of “lumped,” or average, system characteristics in each reactor, such as microbial storage product contents. In reality, the states of individual bacteria are likely to vary due to variable residence times in reactors with completely mixed hydraulics. Earlier work by the present author introduced the MATLAB-based distributed state simulation program, Dissimulator 1.0, and demonstrated that distributed states may be particularly important in enhanced biological phosphorus removal (EBPR) systems, which rely on the cycling of bacteria through anaerobic and aerobic reactors to select for a population accumulating multiple microbial storage products. This paper explores the relationships between distributed state profiles, variable anaerobic and aerobic SRTs, and the process rates predicted by lumped and distributed approaches. Consistent with previous results, the lumped approach consistently predicted better EBPR performance than did the distributed approach. The primary reason for this was the presence of large fractions of polyphosphate accumulating organisms (PAOs) with depleted microbial storage product contents, which led to overestimation of process rates by the lumped approach. Distributed and lumped predictions were therefore most similar when microbial storage product depletion was minimal. The effects of variable anaerobic and aerobic SRTs on distributed profile characteristics and process rates are presented. This work demonstrated that lumped assumptions may overestimate EBPR performance, and the degree of this error is a function of the distributed state profile characteristics such as the degree to which fractions of the biomass contain depleted microbial storage product contents.

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.

Microbial storage products, biomass density, and settling properties of enhanced biological phosphorus removal activated sludge

2001 ◽  
Vol 43 (1) ◽  
pp. 173-180 ◽  
Author(s):  
A. J. Schuler ◽  
D. Jenkins ◽  
P. Ronen
Keyword(s):  
Phosphorus Removal ◽  
P Uptake ◽  
Volume Index ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Biomass Density ◽  
Model Predictions ◽  
Storage Products ◽  
Biological Phosphorus

The relationships between bacterial storage products, density, and settling characteristics were determined in a laboratory-scale sequencing batch reactor (SBR) enhanced biological phosphorus removal (EBPR) system. Both long-term and single anaerobic-aerobic cycle variations in these properties were studied. Increased polyphosphate (PP) content of the biomass during long-term operation resulted in improved sludge volume index (SVI) values. End-aerobic phase (after phosphate (P) uptake) values were consistently lower than end-anaerobic phase (after P release) values. Neither filamentous nor slime bulking were evident by microscopic observations. Biomass density increased at a rate of 1.2 mg/L per each 1% increase in biomass P content. End-aerobic phase samples had an average 25% higher buoyant density than end-anaerobic phase samples, which was attributed to aerobic P uptake. Biomass density was negatively correlated with SVI values, and SVI values increased sharply at low biomass density. A mathematical model developed by Mas et al. (1985) was modified to predict total cell density based on literature values of PP, glycogen (GLY), and poly-b-hydroxybutyrate (PHB) densities. Model predictions were in good agreement with experimental results, although improved measurement of PP density is required to improve model predictions.

Calcium effect on enhanced biological phosphorus removal

2006 ◽  
Vol 53 (12) ◽  
pp. 29-37 ◽  
Author(s):  
R. Barat ◽  
T. Montoya ◽  
L. Borras ◽  
A. Seco ◽  
J. Ferrer
Keyword(s):  
Calcium Phosphate ◽  
Metabolic Pathway ◽  
Phosphorus Removal ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
High Concentration ◽  
P Release ◽  
Calcium Effect ◽  
Biological Phosphorus

The role of calcium (Ca) in enhanced biological phosphorus removal and its possible implications on the metabolic pathway have been studied. The experience has been carried out in an SBR under anaerobic–aerobic conditions for biological phosphorus removal during 8 months. The variations of influent Ca concentration showed a clear influence on the EBPR process, detecting significant changes in YPO4. These YPO4 variations were not due to influent P/COD ratio, pH, denitrification and calcium phosphate formation. The YPO4 has been found to be highly dependent on the Ca concentration, increasing as Ca concentration decreases. The results suggest that high Ca concentrations produce “inert” granules of polyphosphate with Ca as a counterion that are not involved in P release and uptake. Furthermore, microbiological observations confirmed that appreciable changes in PAO and GAO populations were not observed. This behaviour could suggest a change in the bacterial metabolic pathway, with prevailing polyphosphate-accumulating metabolism (PAM) at low influent Ca concentration and glycogen-accumulating metabolism (GAM) at high concentration.

Heterotrophic Kinetic Parameter Estimation for Enhanced Biological Phosphorus Removal Processes Operated in Conventional and Membrane-Assisted Modes

2006 ◽  
Vol 41 (1) ◽  
pp. 72-83 ◽  
Author(s):  
Zhe Zhang ◽  
Eric R. Hall
Keyword(s):  
Parameter Estimation ◽  
Phosphorus Removal ◽  
Growth Yield ◽  
Pilot Scale ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Nitrogen And Phosphorus ◽  
Parameter Values ◽  
The Impact ◽  
Biological Phosphorus

Abstract Parameter estimation and wastewater characterization are crucial for modelling of the membrane enhanced biological phosphorus removal (MEBPR) process. Prior to determining the values of a subset of kinetic and stoichiometric parameters used in ASM No. 2 (ASM2), the carbon, nitrogen and phosphorus fractions of influent wastewater at the University of British Columbia (UBC) pilot plant were characterized. It was found that the UBC wastewater contained fractions of volatile acids (SA), readily fermentable biodegradable COD (SF) and slowly biodegradable COD (XS) that fell within the ASM2 default value ranges. The contents of soluble inert COD (SI) and particulate inert COD (XI) were somewhat higher than ASM2 default values. Mixed liquor samples from pilot-scale MEBPR and conventional enhanced biological phosphorus removal (CEBPR) processes operated under parallel conditions, were then analyzed experimentally to assess the impact of operation in a membrane-assisted mode on the growth yield (YH), decay coefficient (bH) and maximum specific growth rate of heterotrophic biomass (µH). The resulting values for YH, bH and µH were slightly lower for the MEBPR train than for the CEBPR train, but the differences were not statistically significant. It is suggested that MEBPR simulation using ASM2 could be accomplished satisfactorily using parameter values determined for a conventional biological phosphorus removal process, if MEBPR parameter values are not available.

Full Scale Investigations on Enhanced Biological Phosphorus Removal – P-Release in the Anaerobic Reactor

1994 ◽  
Vol 29 (7) ◽  
pp. 153-156 ◽  
Author(s):  
D. Wedi ◽  
P. A. Wilderer
Keyword(s):  
Phosphorus Removal ◽  
Full Scale ◽  
Process Conditions ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Anaerobic Reactor ◽  
P Release ◽  
Acinetobacter Sp ◽  
Biological Phosphorus ◽  
P Removal

Most of the fundamental processes responsible for enhanced biological phosphorus removal (EBPR) were obtained through laboratory tests under defined conditions with pure or enriched cultures. Acinetobacter sp. was identified as the most important group of bacteria responsible for bio-P removal. Full scale data showed, however, that laboratory results do not match full scale results well enough. There is a lack of data on the effects of sub-optimal process conditions such as inadequate availability of volatile fatty acids (VFA), high nitrate recycle, storm water inflow or low temperatures. In this paper the results of full scale experiments on P-release are presented and compared with theoretical values. Measurements at a full scale Phoredox-system showed a surprisingly low P-release in the anaerobic reactor. Only 4 to 10% of the phosphorus in the activated sludge was released in the bulk liquid. With laboratory batch-tests, a maximum of 20% of the P in the sludge could be released. It is assumed that under the prevailing process conditions either the fraction of Acinetobacter sp. was very small, or bacteria other than Acinetobacter sp. were responsible for the P-removal, or most of the phosphorus was bound chemically but mediated by biological processes.

A flux-based stoichiometric model of enhanced biological phosphorus removal metabolism

1998 ◽  
Vol 37 (4-5) ◽  
pp. 609-613
Author(s):  
J. Pramanik ◽  
P. L. Trelstad ◽  
J. D. Keasling
Keyword(s):  
Phosphorus Removal ◽  
Reaction Network ◽  
Bacterial Biomass ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Stoichiometric Model ◽  
Aerobic Reactor ◽  
Complete Set ◽  
Biological Phosphorus

Enhanced biological phosphorus removal (EBPR) in wastewater treatment involves metabolic cycling through the biopolymers polyphosphate (polyP), polyhydroxybutyrate (PHB), and glycogen. This cycling is induced through treatment systems that alternate between carbon-rich anaerobic and carbon-poor aerobic reactor basins. While the appearance and disappearance of these biopolymers has been documented, the intracellular pressures that regulate their synthesis and degradation are not well understood. Current models of the EBPR process have examined a limited number of metabolic pathways that are frequently lumped into an even smaller number of “reactions.” This work, on the other hand, uses a stoichiometric model that contains a complete set of the pathways involved in bacterial biomass synthesis and energy production to examine EBPR metabolism. Using the stoichiometric model we were able to analyze the role of EBPR metabolism within the larger context of total cellular metabolism, as well as predict the flux distribution of carbon and energy fluxes throughout the total reaction network. The model was able to predict the consumption of PHB, the degradation of polyP, the uptake of acetate and the release of Pi. It demonstrated the relationship between acetate uptake and Pi release, and the effect of pH on this relationship. The model also allowed analysis of growth metabolism with respect to EBPR.

Sign in / Sign up

Export Citation Format

Share Document