Oligotyping and metagenomics reveal distinct Candidatus Accumulibacter communities in side-stream versus conventional full-scale enhanced biological phosphorus removal (EBPR) systems

2021 ◽  
Vol 206 ◽  
pp. 117725
Author(s):  
Varun N. Srinivasan ◽  
Guangyu Li ◽  
Dongqi Wang ◽  
Nicholas B. Tooker ◽  
Zihan Dai ◽  
...  
2019 ◽  
Author(s):  
Varun N. Srinivasan ◽  
Guangyu Li ◽  
Dongqi Wang ◽  
Nicholas B. Tooker ◽  
Zihan Dai ◽  
...  

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.


Author(s):  
Dongqi Wang ◽  
Nicholas B. Tooker ◽  
Varun Srinivasan ◽  
Guangyu Li ◽  
Peter Schauer ◽  
...  

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.


1994 ◽  
Vol 29 (7) ◽  
pp. 153-156 ◽  
Author(s):  
D. Wedi ◽  
P. A. Wilderer

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.


2014 ◽  
Vol 66 ◽  
pp. 283-295 ◽  
Author(s):  
Ana B. Lanham ◽  
Adrian Oehmen ◽  
Aaron M. Saunders ◽  
Gilda Carvalho ◽  
Per H. Nielsen ◽  
...  

2007 ◽  
Vol 73 (18) ◽  
pp. 5865-5874 ◽  
Author(s):  
Shaomei He ◽  
Daniel L. Gall ◽  
Katherine D. McMahon

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.


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