scholarly journals Oligotyping and Genome-Resolved Metagenomics Reveal DistinctCandidatusAccumulibacter Communities in Full-Scale Side-Stream versus Conventional Enhanced Biological Phosphorus Removal (EBPR) Configurations

2019 ◽  
Author(s):  
Varun N. Srinivasan ◽  
Guangyu Li ◽  
Dongqi Wang ◽  
Nicholas B. Tooker ◽  
Zihan Dai ◽  
...  
Keyword(s):  
16S Rrna ◽  
16S Rrna Gene ◽  
Phosphorus Removal ◽  
Full Scale ◽  
Rrna Gene ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Side Stream ◽  
First Time ◽  
Biological Phosphorus

AbstractCandidatusAccumulibacter phosphatis (CAP) and its sub-clades-level diversity has been associated and implicated in successful phosphorus removal performance in enhanced biological phosphorus removal (EBPR). Development of high-throughput untargeted methods to characterize clades of CAP in EBPR communities can enable a better understanding of Accumulibacter ecology at a higher-resolution beyond OTU-level in wastewater resource recovery facilities (WRRFs). In this study, for the first time, using integrated 16S rRNA gene sequencing, oligotyping and genome-resolved metagenomics, we were able to reveal clade-level differences in Accumulibacter communities and associate the differences with two different full-scale EBPR configurations. The results led to the identification and characterization of a distinct and dominant Accumulibacter oligotype - Oligotype 2 (belonging to Clade IIC) and its matching MAG (RC14) associated with side-stream EBPR configuration. We are also able to extract MAGs belonging to CAP clades IIB (RCAB4-2) and II (RC18) which did not have representative genomes before. This study demonstrates and validates the use of a high-throughput approach of oligotyping analysis of 16S rRNA gene sequences to elucidate CAP clade-level diversity. We also show the existence of a previously uncharacterized diversity of CAP clades in full-scale EBPR communities through extraction of MAGs, for the first time from full-scale facilities.

scholarly journals “Candidatus Accumulibacter” Population Structure in Enhanced Biological Phosphorus Removal Sludges as Revealed by Polyphosphate Kinase Genes

2007 ◽  
Vol 73 (18) ◽  
pp. 5865-5874 ◽  
Author(s):  
Shaomei He ◽  
Daniel L. Gall ◽  
Katherine D. McMahon
Keyword(s):  
Population Structure ◽  
16S Rrna ◽  
Genetic Marker ◽  
Phosphorus Removal ◽  
Full Scale ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Biological Phosphorus

ABSTRACT We investigated the fine-scale population structure of the “Candidatus Accumulibacter” lineage in enhanced biological phosphorus removal (EBPR) systems using the polyphosphate kinase 1 gene (ppk1) as a genetic marker. We retrieved fragments of “Candidatus Accumulibacter” 16S rRNA and ppk1 genes from one laboratory-scale and several full-scale EBPR systems. Phylogenies reconstructed using 16S rRNA genes and ppk1 were largely congruent, with ppk1 granting higher phylogenetic resolution and clearer tree topology and thus serving as a better genetic marker than 16S rRNA for revealing population structure within the “Candidatus Accumulibacter” lineage. Sequences from at least five clades of “Candidatus Accumulibacter” were recovered by ppk1-targeted PCR, and subsequently, specific primer sets were designed to target the ppk1 gene for each clade. Quantitative real-time PCR (qPCR) assays using “Candidatus Accumulibacter”-specific 16S rRNA and “Candidatus Accumulibacter” clade-specific ppk1 primers were developed and conducted on three laboratory-scale and nine full-scale EBPR samples and two full-scale non-EBPR samples to determine the abundance of the total “Candidatus Accumulibacter” lineage and the relative distributions and abundances of the five “Candidatus Accumulibacter” clades. The qPCR-based estimation of the total “Candidatus Accumulibacter” fraction as a proportion of the bacterial community as measured using 16S rRNA genes was not significantly different from the estimation measured using ppk1, demonstrating the power of ppk1 as a genetic marker for detection of all currently defined “Candidatus Accumulibacter” clades. The relative distributions of “Candidatus Accumulibacter” clades varied among different EBPR systems and also temporally within a system. Our results suggest that the “Candidatus Accumulibacter” lineage is more diverse than previously realized and that different clades within the lineage are ecologically distinct.

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 Global warming readiness: Feasibility of enhanced biological phosphorus removal from wastewater at 35°C

2021 ◽  
Author(s):  
Guanglei Qiu ◽  
Yingyu Law ◽  
Rogelio Zuniga-Montanez ◽  
Yang Lu ◽  
Samarpita Roy ◽  
...  
Keyword(s):  
Phosphorus Removal ◽  
Municipal Wastewater ◽  
Feeding Strategy ◽  
Full Scale ◽  
Carbon Uptake ◽  
Rrna Gene ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Biological Phosphorus ◽  
P Removal

AbstractRecent research has shown enhanced biological phosphorus removal (EBPR) from municipal wastewater at warmer temperatures around 30°C to be stable in both laboratory-scale reactors and full-scale treatment plants. In the context of a changing climate, the feasibility of EBPR at even higher temperatures is of interest. We operated two lab-scale EBPR sequencing batch reactors with alternating anaerobic and aerobic phases for over 300 days at 30°C and 35°C, respectively, and followed the dynamics of the communities of phosphorus accumulating organisms (PAOs) and competing glycogen accumulating organisms (GAOs) using a combination of 16S rRNA gene metabarcoding, quantitative PCR and fluorescent in-situ hybridization analyses. Stable and nearly complete P removal was achieved at 30°C; similarly, long term P removal was stable at 35°C with effluent PO43−-P concentrations < 0.5 mg/L on half of all monitored days. Diverse and abundant Ca. Accumulibacter amplicon sequence variants were closely related to those found in temperate environments, suggesting that EBPR at this temperature does not require a highly specialized PAO community. The slow-feeding strategy used effectively limited the carbon uptake rates of GAOs, allowing PAOs to outcompete GAOs at both temperatures. Candidatus Competibacter was the main GAO, along with cluster III Defluviicoccus members. These organisms withstood the slow-feeding regime, suggesting that their bioenergetic characteristics of carbon uptake differ from those of their tetrad-forming relatives. This specific lineage of GAOs warrants further study to establish how complete P removal can be maintained. Comparative cycle studies at two temperatures for each reactor revealed higher activity of Ca. Accumulibacter when the temperature was increased from 30°C to 35°C, suggesting that the stress was a result of the higher carbon (and/or P) metabolic rates of PAOs and GAOs, the resultant carbon deficiency, and additional community competition. An increase in the TOC to PO43--P ratio (from 25:1 to 40:1) effectively eased the carbon deficiency and benefited the proliferation of PAOs. In general, the slow-feeding strategy and sufficiently high carbon input benefited a high and stable EBPR at elevated temperature and represent basic conditions for full-scale applications.

scholarly journals A Full-Scale Comparative Study of Conventional and Side-Stream Enhanced Biological Phosphorus Removal Processes

2018 ◽  
Author(s):  
Dongqi Wang ◽  
Nicholas B. Tooker ◽  
Varun Srinivasan ◽  
Guangyu Li ◽  
Peter Schauer ◽  
...  
Keyword(s):  
Phosphorus Removal ◽  
Tca Cycle ◽  
Full Scale ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Solid Retention Time ◽  
Side Stream ◽  
Significant Difference ◽  
Polyphosphate Accumulating Organisms ◽  
Biological Phosphorus

In this study, a full-scale pilot testing was performed with side-by-side operation of a conventional enhanced biological phosphorus removal (EBPR) process and a side-stream EBPR (S2EBPR) process. A comparison of the performance, activities and population dynamics of key functionally relevant populations between the two configurations were carried out. The results demonstrated that, with the same influent wastewater characteristics, S2EBPR configuration showed more effective and stable orthophosphate (PO4-P) removal performance (up to 94% with average effluent concentration down to 0.1 mg P/L) than conventional EBPR, especially when the mixers in side-stream reactor were operated intermittently. Mass balance analysis illustrated that both denitrification and EBPR performance have been enhanced in S2EBPR configuration through diverting primary effluent to anoxic zone and producing additional carbon (~40%) via fermentation in side-stream reactor. Microbial characterization showed that there was no significant difference in the relative abundances of Ca. Accumulibacter (~5.9%) and Tetrasphaera (~16%) putative polyphosphate-accumulating organisms (PAOs) between the two configurations. However, lower relative abundance of known GAOs was observed in S2EBPR configuration (1.1%) than the conventional one (2.7%). A relatively higher PAO activity and increased degree of dependence on glycolysis pathway than TCA cycle was observed in S2EBPR configuration using P release and uptake batch test. Adequate anaerobic solid retention time (SRT) and conditions that generate continuous and slow feeding/production of volatile fatty acid (VFA) with higher composition percentage of propionate in the side-stream reactor of S2EBPR process likely provide a competitive advantage for PAOs over GAOs.

Fine-scale differences between Accumulibacter-like bacteria in enhanced biological phosphorus removal activated sludge

2006 ◽  
Vol 54 (1) ◽  
pp. 111-117 ◽  
Author(s):  
S. He ◽  
A.Z. Gu ◽  
K.D. McMahon
Keyword(s):  
Phosphorus Removal ◽  
Wastewater Treatment Plants ◽  
Batch Reactor ◽  
Comparative Sequence Analysis ◽  
Full Scale ◽  
Rrna Gene ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Phylogenetic Resolution ◽  
Biological Phosphorus

A lab-scale sequencing batch reactor (SBR) and six full-scale wastewater treatment plants (WWTPs) performing enhanced biological phosphorus removal (EBPR) were surveyed. The abundance of Accumulibacter-related organisms in the full-scale plants was investigated using fluorescent in situ hybridization. Accumulibacter-related organisms were present in all of the full-scale EBPR plants, at levels ranging from 9% to 24% of total cells. The high percentage of Accumulibacter-related organisms seemed to be associated with configurations which minimize the nitrate recycling to the anaerobic zone and low influent BOD:TP ratios. PCR-based clone libraries were constructed from the community 16S rRNA gene plus the internally transcribed spacer region amplified from the SBR and five of the full-scale WWTPs. Comparative sequence analysis was carried out using Accumulibacter-related clones, providing higher phylogenetic resolution and revealing finer-scale clustering of the sequences retrieved from the SBR and full-scale EBPR plants.

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.

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