Microbial community analysis involved in the aerobic/extended-idle process performing biological phosphorus removal

2013 ◽  
Vol 67 (3) ◽  
pp. 485-493 ◽  
Author(s):  
Tian-jing Zeng ◽  
Guo-jing Yang ◽  
Dong-bo Wang ◽  
Xiao-ming Li ◽  
Wei Zheng ◽  
...  
Keyword(s):  
Phosphorus Removal ◽  
Community Analysis ◽  
Microbial Community Analysis ◽  
Fish Analysis ◽  
Biological Phosphorus Removal ◽  
Acclimation Period ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus ◽  
Total Bacteria

Recently, it has been found that biological phosphorus removal can be achieved in an aerobic/extended-idle (AEI) process using both glucose and acetate as the sole substrate. However, the microbial consortiums involved in glucose-fed and acetate-fed systems have not yet been characterized. Thus the aims of this paper were to investigate the diversities and dynamics of bacterial communities during the acclimation period, and to quantify polyphosphate-accumulating organisms (PAOs) and glycogen-accumulating organisms (GAOs) in the systems. The phylogenetic analysis showed that the microbial communities were mainly composed of phylum Proteobacteria, Bacteroidetes, Chlorobi and another six kinds of unclassified bacteria. Fluorescence in-situ hybridization (FISH) analysis revealed that PAOs and GAOs accounted for 43 ± 7 and 16 ± 3% of all bacteria in the glucose-fed system, and 19 ± 4 and 35 ± 5% of total bacteria in the acetate-fed system, respectively. The results showed that the conventional PAOs could thrive in the AEI process, and a defined anaerobic zone was not necessarily required for putative PAOs growth.

Presence of Rhodocyclus in a full-scale wastewater treatment plant and their participation in enhanced biological phosphorus removal

2002 ◽  
Vol 46 (1-2) ◽  
pp. 123-128 ◽  
Author(s):  
J.L. Zilles ◽  
C.-H. Hung ◽  
D.R. Noguera
Keyword(s):  
Wastewater Treatment ◽  
Phosphorus Removal ◽  
Fluorescent In Situ Hybridization ◽  
Wastewater Treatment Plants ◽  
Full Scale ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

The objective of this research was to assess the relevance of organisms related to Rhodocyclus in enhanced biological phosphorus removal in full-scale wastewater treatment plants. The presence of these organisms in full-scale plants was first confirmed by fluorescent in situ hybridization. To address which organisms were involved in phosphorus removal, a method was developed which selected polyphosphate-accumulating organisms from activated sludge samples by DAPI staining and flow cytometry. Sorted samples were characterized using fluorescent in situ hybridization. The results of these analyses confirmed the presence of organisms related to Rhodocyclus in full-scale wastewater treatment plants and supported the involvement of these organisms in enhanced biological phosphorus removal. However, a significant fraction of the polyphosphate-accumulating organisms were not related to Rhodocyclus.

The effect of GAOs (glycogen accumulating organisms) on anaerobic carbon requirements in full-scale Australian EBPR (enhanced biological phosphorus removal) plants

2003 ◽  
Vol 47 (11) ◽  
pp. 37-43 ◽  
Author(s):  
A.M. Saunders ◽  
A. Oehmen ◽  
L.L. Blackall ◽  
Z. Yuan ◽  
J. Keller
Keyword(s):  
Phosphorus Removal ◽  
In Situ Hybridisation ◽  
Full Scale ◽  
Fluorescence In Situ Hybridisation ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Accumulibacter Phosphatis ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

Glycogen-accumulating organisms (GAOs) were present in six full-scale plants investigated and in all but one made a significant contribution to the amount of volatile fatty acid (VFA) taken up anaerobically. While most plants surveyed contain GAOs, it was demonstrated that it is possible for a full-scale plant to operate with an insignificant GAO population.“Candidatus Accumulibacter phosphatis”were the significant polyphosphate-accumulating organisms (PAOs) in all plants surveyed. “Candidatus Competibacter phosphatis” were found in all plants along with other possible GAOs that were observed but not identified. A significant GAO population will increase the carbon requirements by removing VFA that could otherwise have been used by PAOs. Process optimization minimizing GAOs in full-scale plants would lead to a more efficient use of VFA. Enhanced biological phosphorus removal (EBPR), fluorescence in situ hybridisation (FISH), glycogen accumulating organism (GAO); polyphosphate accumulating organism (PAO);

scholarly journals Identity and Ecophysiology of Uncultured Actinobacterial Polyphosphate-Accumulating Organisms in Full-Scale Enhanced Biological Phosphorus Removal Plants

2005 ◽  
Vol 71 (7) ◽  
pp. 4076-4085 ◽  
Author(s):  
Yunhong Kong ◽  
Jeppe Lund Nielsen ◽  
Per Halkjær Nielsen
Keyword(s):  
Phosphorus Removal ◽  
Treatment Plant ◽  
Full Scale ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Fish Analysis ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

ABSTRACT Microautoradiography combined with fluorescence in situ hybridization (MAR-FISH) was used to screen for potential polyphosphate-accumulating organisms (PAO) in a full-scale enhanced biological phosphorus removal (EBPR) plant. The results showed that, in addition to uncultured Rhodocyclus-related PAO, two morphotypes hybridizing with gene probes for the gram-positive Actinobacteria were also actively involved in uptake of orthophosphate (Pi). Clone library analysis and further investigations by MAR-FISH using two new oligonucleotide probes revealed that both morphotypes, cocci in clusters of tetrads and short rods in clumps, were relatively closely related to the genus Tetrasphaera within the family Intrasporangiaceae of the Actinobacteria (93 to 98% similarity in their 16S rRNA genes). FISH analysis of the community biomass in the treatment plant investigated showed that the short rods (targeted by probe Actino-658) were the most abundant (12% of all Bacteria hybridizing with general bacterial probes), while the cocci in tetrads (targeted by probe Actino-221) made up 7%. Both morphotypes took up Pi aerobically only if, in a previous anaerobic phase, they had taken up organic matter from wastewater or a mixture of amino acids. They could not take up short-chain fatty acids (e.g., acetate), glucose, or ethanol under anaerobic or aerobic conditions. The storage compound produced during the anaerobic period was not polyhydroxyalkanoates, as for Rhodocyclus-related PAO, and its identity is still unknown. Growth and uptake of Pi took place in the presence of oxygen and nitrate but not nitrite, indicating a lack of denitrifying ability. A survey of the occurrence of these actinobacterial PAO in 10 full-scale EBPR plants revealed that both morphotypes were widely present, and in several plants more abundant than the Rhodocyclus-related PAO, thus playing a very important role in the EBPR process.

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.

scholarly journals Resolving the individual contribution of key microbial populations to enhanced biological phosphorus removal with Raman-FISH

2018 ◽  
Author(s):  
Eustace Y. Fernando ◽  
Simon Jon Mcllroy ◽  
Marta Nierychlo ◽  
Florian-Alexander Herbst ◽  
Markus C. Schmid ◽  
...  
Keyword(s):  
Phosphorus Removal ◽  
Absolute Quantification ◽  
Full Scale ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Important Player ◽  
The Individual ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

AbstractEnhanced biological phosphorus removal (EBPR) is a globally important biotechnological process and relies on the massive accumulation of phosphate within special microorganisms.CandidatusAccumulibacter conform to classical physiology model for polyphosphate accumulating organisms and are widely believed to be the most important player for the process in full-scale EBPR systems. However, it was impossible till now to quantify the contribution of specific microbial clades to EBPR. In this study, we have developed a new tool to directly link the identity of microbial cells to the absolute quantification of intracellular poly-P and other polymers underin situconditions, and applied it to eight full-scale EBPR plants. BesidesCa. Accumulibacter, members of the genusTetrasphaerawere found to be important microbes for P accumulation, and in six plants they were the most important. As theseTetrasphaeracells did not exhibit the classical phenotype of poly-P accumulating microbes, our entire understanding of the microbiology of the EBPR process has to be revised. Furthermore, our new single-cell approach can now also be applied to quantify storage polymer dynamics in individual populationsin situin other ecosystems and might become a valuable tool for many environmental microbiologists.

scholarly journals Microautoradiographic Study of Rhodocyclus-Related Polyphosphate-Accumulating Bacteria in Full-Scale Enhanced Biological Phosphorus Removal Plants

2004 ◽  
Vol 70 (9) ◽  
pp. 5383-5390 ◽  
Author(s):  
Yunhong Kong ◽  
Jeppe Lund Nielsen ◽  
Per Halkjær Nielsen
Keyword(s):  
Electron Acceptor ◽  
Phosphorus Removal ◽  
Tricarboxylic Acid Cycle ◽  
Reducing Power ◽  
Full Scale ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

ABSTRACT The ecophysiology of uncultured Rhodocyclus-related polyphosphate-accumulating organisms (PAO) present in three full-scale enhanced biological phosphorus removal (EBPR) activated sludge plants was studied by using microautoradiography combined with fluorescence in situ hybridization. The investigations showed that these organisms were present in all plants examined and constituted 5 to 10, 10 to 15, and 17 to 22% of the community biomass. The behavior of these bacteria generally was consistent with the biochemical models proposed for PAO, based on studies of lab-scale investigations of enriched and often unknown PAO cultures. Rhodocyclus-related PAO were able to accumulate short-chain substrates, including acetate, propionate, and pyruvate, under anaerobic conditions, but they could not assimilate many other low-molecular-weight compounds, such as ethanol and butyrate. They were able to assimilate two substrates (e.g., acetate and propionate) simultaneously. Leucine and thymidine could not be assimilated as sole substrates and could only be assimilated as cosubstrates with acetate, perhaps serving as N sources. Glucose could not be assimilated by the Rhodocyclus-related PAO, but it was easily fermented in the sludge to products that were subsequently consumed. Glycolysis, and not the tricarboxylic acid cycle, was the source that provided the reducing power needed by the Rhodocyclus-related PAO to form the intracellular polyhydroxyalkanoate storage compounds during anaerobic substrate assimilation. The Rhodocyclus-related PAO were able to take up orthophosphate and accumulate polyphosphate when oxygen, nitrate, or nitrite was present as an electron acceptor. Furthermore, in the presence of acetate growth was sustained by using oxygen, as well as nitrate or nitrite, as an electron acceptor. This strongly indicates that Rhodocyclus-related PAO were able to denitrify and thus played a role in the denitrification occurring in full-scale EBPR plants.

scholarly journals Population Structure and Morphotype Analysis of “CandidatusAccumulibacter” Using FluorescenceIn SituHybridization-Staining-Flow Cytometry

2019 ◽  
Vol 85 (9) ◽  
Author(s):  
Chao Li ◽  
Wei Zeng ◽  
Ning Li ◽  
Yu Guo ◽  
Yongzhen Peng
Keyword(s):  
Flow Cytometry ◽  
Population Structure ◽  
Electron Acceptor ◽  
Phosphorus Removal ◽  
Fish Analysis ◽  
Bacterial Cells ◽  
Denitrifying Phosphorus Removal ◽  
Optimal Method ◽  
Total Bacteria

ABSTRACT“CandidatusAccumulibacter” is the dominant polyphosphate-accumulating organism (PAO) in denitrifying phosphorus removal (DPR) systems. In order to investigate the community structure and clade morphotypes of “CandidatusAccumulibacter” in DPR systems through flow cytometry (FCM), denitrifying phosphorus removal of almost 100% using nitrite and nitrate as the electron acceptor was achieved in sequencing batch reactors (SBRs). An optimal method of flow cytometry combined with fluorescencein situhybridization and SYBR green I staining (FISH-staining-flow cytometry) was developed to quantify PAOs in DPR systems. By setting the width value of FCM, bacterial cells in a sludge sample were divided into three groups in different morphotypes, namely, coccus, coccobacillus, and bacillus. Average percentages that the three different PAO populations accounted for among total bacteria from SBR1 (SBR2) were 42% (45%), 14% (13%), and 4% (2%). FCM showed that the ratios of PAOs to total bacteria in the two reactors were 61% and 59%, and the quantitative PCR (qPCR) results indicated that IIC was the dominant “CandidatusAccumulibacter” clade in both denitrifying phosphorus removal systems, reaching 50% of the total “CandidatusAccumulibacter” bacteria. The subdominant clade in the reactor with nitrite as the electron acceptor was IID, accounting for 31% of the total “CandidatusAccumulibacter” bacteria. The FCM and qPCR results suggested that clades IIC and IID were both coccus, clade IIF was coccobacillus, and clade IA was bacillus. FISH analysis also indicated that PAOs were major cocci in the systems. An equivalence test of FCM-based quantification confirmed the accuracy of FISH-staining-flow cytometry, which can meet the quantitative requirements for PAOs in complex activated sludge samples.IMPORTANCEAs one group of the most important functional phosphorus removal organisms, “CandidatusAccumulibacter,” affiliated with theRhodocyclusgroup of theBetaproteobacteria, is a widely recognized and studied PAO in the field of biological wastewater treatment. The morphotypes and population structure of clade-level “CandidatusAccumulibacter” were studied through novel FISH-staining-flow cytometry, which involved denitrifying phosphorus removal (DPR) achieving carbon and energy savings and simultaneous removal of N and P, thus inferring the different denitrifying phosphorus removal abilities of these clades. Additionally, based on this method,in situquantification for specific polyphosphate-accumulating organisms (PAOs) enables a more efficient process and more accurate result. The establishment of FISH-staining-flow cytometry makes cell sorting of clade-level noncultivated organisms available.

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.

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