scholarly journals Influence of Extraction Solvent on Nontargeted Metabolomics Analysis of Enrichment Reactor Cultures Performing Enhanced Biological Phosphorus Removal (EBPR)

Metabolites ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 269
Author(s):  
Nay Min Min Thaw Saw ◽  
Pipob Suwanchaikasem ◽  
Rogelio Zuniga-Montanez ◽  
Guanglei Qiu ◽  
Ezequiel M. Marzinelli ◽  
...  
Keyword(s):  
Microbial Communities ◽  
Extraction Methods ◽  
Ultra Performance Liquid Chromatography ◽  
Phosphate Removal ◽  
Solvent Mixtures ◽  
Biological Phosphorus Removal ◽  
Polar Solvents ◽  
Extraction Solvent ◽  
Polyphosphate Accumulating Organisms ◽  
Metabolomics Analysis

Metabolome profiling is becoming more commonly used in the study of complex microbial communities and microbiomes; however, to date, little information is available concerning appropriate extraction procedures. We studied the influence of different extraction solvent mixtures on untargeted metabolomics analysis of two continuous culture enrichment communities performing enhanced biological phosphate removal (EBPR), with each enrichment targeting distinct populations of polyphosphate-accumulating organisms (PAOs). We employed one non-polar solvent and up to four polar solvents for extracting metabolites from biomass. In one of the reactor microbial communities, we surveyed both intracellular and extracellular metabolites using the same set of solvents. All samples were analysed using ultra-performance liquid chromatography mass spectrometry (UPLC-MS). UPLC-MS data obtained from polar and non-polar solvents were analysed separately and evaluated using extent of repeatability, overall extraction capacity and the extent of differential abundance between physiological states. Despite both reactors demonstrating the same bioprocess phenotype, the most appropriate extraction method was biomass specific, with methanol: water (50:50 v/v) and methanol: chloroform: water (40:40:20 v/v) being chosen as the most appropriate for each of the two different bioreactors, respectively. Our approach provides new data on the influence of solvent choice on the untargeted surveys of the metabolome of PAO enriched EBPR communities and suggests that metabolome extraction methods need to be carefully tailored to the specific complex microbial community under study.

scholarly journals Evaluation of extraction solvents for untargeted metabolomics analysis of enrichment reactor cultures performing enhanced biological phosphorus removal (EBPR)

2019 ◽  
Author(s):  
Nay Min Min Thaw Saw ◽  
Pipob Suwanchaikasem ◽  
Rogelio Zuniga-Montanez ◽  
Guanglei Qiu ◽  
Stefan Wuertz ◽  
...  
Keyword(s):  
Phosphorus Removal ◽  
Extraction Methods ◽  
Untargeted Metabolomics ◽  
Solvent Mixtures ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Extraction Solvent ◽  
Analytical Protocol ◽  
Biological Phosphorus ◽  
Metabolomics Analysis

AbstractIntroductionThe extraction solvent mixtures were optimized for untargeted metabolomics analysis of microbial communities from two laboratory scale activated sludge reactors performing enhanced biological phosphorus removal (EBPR).ObjectiveTo develop a robust and simple analytical protocol to analyse microbial metabolomics from EBPR bioreactors.MethodsExtra- and intra-cellular metabolites were extracted using five methods and analysed by ultraperformance liquid chromatography mass spectrometry (UPLC-MS).ResultsThe optimal extraction method was biomass specific and methanol:water (1:1 v/v) and methanol:chloroform:water (2:2:1 v/v) were chosen, respectively, for each of the two different bioreactors.ConclusionOur approach provides direct surveys of the metabolic state of PAO-enriched EBPR communities, showing that extraction methods should be carefully tailored to the microbial community under study

scholarly journals “Candidatus Dechloromonas phosphoritropha” and “Ca. D. phosphorivorans”, novel polyphosphate accumulating organisms abundant in wastewater treatment systems

2021 ◽  
Author(s):  
Francesca Petriglieri ◽  
Caitlin Singleton ◽  
Miriam Peces ◽  
Jette F. Petersen ◽  
Marta Nierychlo ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Niche Partitioning ◽  
16S Rrna Gene Sequencing ◽  
Abundant Species ◽  
Phosphate Removal ◽  
Rrna Gene ◽  
Biological Phosphorus Removal ◽  
Polyphosphate Accumulating Organisms

AbstractMembers of the genus Dechloromonas are often abundant in enhanced biological phosphorus removal (EBPR) systems and are recognized putative polyphosphate accumulating organisms (PAOs), but their role in phosphate removal is still unclear. Here, we used 16S rRNA gene sequencing and fluorescence in situ hybridization (FISH) to investigate the abundance and distribution of Dechloromonas spp. in Danish and global wastewater treatment plants. The two most abundant species worldwide revealed in situ dynamics of important intracellular storage polymers, measured by FISH-Raman in activated sludge from four full-scale EBPR plants and from a lab-scale reactor fed with different substrates. Moreover, seven distinct Dechloromonas species were determined from a set of ten high-quality metagenome-assembled genomes (MAGs) from Danish EBPR plants, each encoding the potential for polyphosphate (poly-P), glycogen, and polyhydroxyalkanoates (PHA) accumulation. The two species exhibited an in situ phenotype in complete accordance with the metabolic information retrieved by the MAGs, with dynamic levels of poly-P, glycogen, and PHA during feast-famine anaerobic–aerobic cycling, legitimately placing these microorganisms among the important PAOs. They are potentially involved in denitrification showing niche partitioning within the genus and with other important PAOs. As no isolates are available for the two species, we propose the names Candidatus Dechloromonas phosphoritropha and Candidatus Dechloromonas phosphorivorans.

scholarly journals Recovery of Phosphorus in Wastewater in the Form of Polyphosphates: A Review

Processes ◽  
2022 ◽  
Vol 10 (1) ◽  
pp. 144
Author(s):  
Weiran Chu ◽  
Yi Shi ◽  
Liang Zhang
Keyword(s):  
Phosphorus Removal ◽  
Renewable Resource ◽  
Phosphate Removal ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
Treatment Processes ◽  
Real Wastewater ◽  
Polyphosphate Accumulating Organisms ◽  
Chain Lengths ◽  
Biological Phosphorus

As non-renewable resource, the recovery and utilization of phosphorus from wastewater is an enduring topic. Stimulated by the advances in research on polyphosphates (polyP) as well as the development of Enhanced Biological Phosphorus Removal (EBPR) technology to achieve the efficient accumulation of polyP via polyphosphate accumulating organisms (PAOs), a novel phosphorus removal strategy is considered with promising potential for application in real wastewater treatment processes. This review mainly focuses on the mechanism of phosphorus aggregation in the form of polyP during the phosphate removal process. Further discussion about the reuse of polyP with different chain lengths is provided herein so as to suggest possible application pathways for this biosynthetic product.

Metal and phosphate speciation during anaerobic digestion of phosphorus rich sludge

1997 ◽  
Vol 36 (6-7) ◽  
pp. 191-200 ◽  
Author(s):  
C. M. Carliell ◽  
A. D. Wheatley
Keyword(s):  
Anaerobic Digestion ◽  
Metal Speciation ◽  
Extraction Methods ◽  
Phosphate Removal ◽  
Metal Species ◽  
Chemical Extraction ◽  
Inorganic P ◽  
Similar Work ◽  
Extractable P ◽  
Phosphate Speciation

Chemical extraction methods are used to investigate metal and phosphate speciation during anaerobic digestion of phosphorus-rich sludge. Tests were performed using model compounds to evaluate the efficacy of the reagents in the extraction sequences and these results compared with similar work by other researchers. The metal speciation method was found to be suitable for identifying shifts in metal distribution but was unrepresentative of actual metal species. The phosphate speciation method did give adequate separation of the phosphate compounds tested. Full-scale digesters treating chemical and biological phosphate removal (CPR and BPR) sludge were analysed according to the methods developed. Results show that digestion of CPR sludge did not increase the soluble P concentration in the digester and that most of the precipitated phosphorus appeared to be retained in the sludge as inorganic P. The digester treating BPR sludge showed increased soluble and water-extractable P, in comparison to the control digester. Trace metal speciation profiles were found to be affected by addition of CPR sludge.

Short-term effects of carbon source on the competition of polyphosphate accumulating organisms and glycogen accumulating organisms

2004 ◽  
Vol 50 (10) ◽  
pp. 139-144 ◽  
Author(s):  
A. Oehmen ◽  
Z. Yuan ◽  
L.L. Blackall ◽  
J. Keller
Keyword(s):  
Carbon Source ◽  
In Situ Hybridisation ◽  
Operating Conditions ◽  
Fluorescence In Situ Hybridisation ◽  
Biological Phosphorus Removal ◽  
Short Term ◽  
Uptake Rates ◽  
Polyphosphate Accumulating Organisms ◽  
The Impact ◽  
Short Term Effects

The effectiveness of enhanced biological phosphorus removal (EBPR) systems is directly affected by the competition of polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs). This study investigated the short-term effects of carbon source on PAO and GAO performance. The tests were designed to clearly determine the impact of volatile fatty acid (VFA) composition on the performance of two types of biomass, one enriched for PAOs and the other for GAOs. The two populations were enriched in separate reactors using identical operating conditions and very similar influent compositions with acetate as the sole carbon source. The only difference was that a very low level of phosphorus was present in the influent to the GAO reactor. The abundance of PAOs and GAOs was quantified using fluorescence in-situ hybridisation. The results clearly show that there are some very distinctive differences between PAOs and GAOs in their ability to utilise different carbon substrates. While both are able to take up acetate rapidly and completely, the GAOs are far slower at consuming propionate than the PAOs during short-term substrate changes. This provides a potentially highly valuable avenue to influence the competition between PAOs and GAOs. Other VFAs studied seem to be less usable in the short term by both PAOs and GAOs, as indicated by their much lower uptake rates.

scholarly journals Assessing high-throughput environmental DNA extraction methods for meta-barcode characterization of aquatic microbial communities

2018 ◽  
Vol 17 (1) ◽  
pp. 37-49 ◽  
Author(s):  
Abdolrazagh Hashemi Shahraki ◽  
Subba Rao Chaganti ◽  
Daniel Heath
Keyword(s):  
Microbial Community ◽  
Microbial Communities ◽  
High Throughput ◽  
Environmentally Friendly ◽  
Environmental Dna ◽  
Magnetic Bead ◽  
Extraction Methods ◽  
Environmental Research ◽  
Rrna Gene

Abstract The characterization of microbial community dynamics using genomic methods is rapidly expanding, impacting many fields including medical, ecological, and environmental research and applications. One of the biggest challenges for such studies is the isolation of environmental DNA (eDNA) from a variety of samples, diverse microbes, and widely variable community compositions. The current study developed environmentally friendly, user safe, economical, and high throughput eDNA extraction methods for mixed aquatic microbial communities and tested them using 16 s rRNA gene meta-barcoding. Five different lysis buffers including (1) cetyltrimethylammonium bromide (CTAB), (2) digestion buffer (DB), (3) guanidinium isothiocyanate (GITC), (4) sucrose lysis (SL), and (5) SL-CTAB, coupled with four different purification methods: (1) phenol-chloroform-isoamyl alcohol (PCI), (2) magnetic Bead-Robotic, (3) magnetic Bead-Manual, and (4) membrane-filtration were tested for their efficacy in extracting eDNA from recreational freshwater samples. Results indicated that the CTAB-PCI and SL-Bead-Robotic methods yielded the highest genomic eDNA concentrations and succeeded in detecting the core microbial community including the rare microbes. However, our study recommends the SL-Bead-Robotic eDNA extraction protocol because this method is safe, environmentally friendly, rapid, high-throughput and inexpensive.

Bacteria and Protozoa Population Dynamics in Biological Phosphate Removal Systems

1994 ◽  
Vol 29 (7) ◽  
pp. 109-117 ◽  
Author(s):  
J. S. Čech ◽  
P. Hartman ◽  
M. Macek
Keyword(s):  
Population Dynamics ◽  
Phosphorus Removal ◽  
Sequencing Batch Reactor ◽  
Batch Reactor ◽  
Sole Source ◽  
Phosphate Removal ◽  
Enhanced Biological Phosphorus Removal ◽  
Biological Phosphorus Removal ◽  
System Collapse ◽  
Biological Phosphorus

Population dynamics of polyphosphate-accumulating bacteria (PP bacteria) was studied in a laboratory sequencing batch reactor simulating anaerobic-oxic sludge system. The competition between PP bacteria and another microorganism (“G bacteria”) for anaerobic-oxic utilization of acetate as the sole source of organic carbon was observed. The competition was found to be seriously influenced by protozoan and metazoan grazing: Predation-resistant “G bacteria” forming large compact flocs outcompeted PP bacteria. Several breakdowns of enhanced biological phosphorus removal were observed. The first one was related to the development of an euglenid flagellate Entosiphon sulcatus and attached ciliates Vorticella microstoma and V. campanula. The second system collapse was connected with a rapid proliferation of rotifers. An alternative-prey predation was thought to be a mechanism of PP bacteria elimination.

Effect of extraction method on phenolic content and antioxidant activity of mistletoe extracts from Viscum album subsp. abietis

2014 ◽  
Vol 68 (7) ◽  
Author(s):  
Wioleta Pietrzak ◽  
Renata Nowak ◽  
Marta Olech
Keyword(s):  
Antioxidant Activity ◽  
Extraction Efficiency ◽  
Antioxidant Activities ◽  
Total Content ◽  
Viscum Album ◽  
Extraction Methods ◽  
Solvent Mixtures ◽  
Ultrasound Assisted Extraction ◽  
Extraction Technology ◽  
Considerable Saving

AbstractThe total content of polyphenols and flavonoids determined in the same plant and their corresponding antioxidant activities may vary widely, depending on the extraction conditions applied. This study was conducted to optimise the extraction conditions of phenolics and flavonoids from the mistletoe plant. Various extraction methods, i.e. ultrasound-assisted extraction technology, maceration, maceration with stirring, accelerated solvent extraction (ASE), and extraction under reflux were evaluated for their percentage extraction of polyphenols (TPC) and flavonoids (TFC) from Viscum album subsp. abietis. In addition, the anti-radical activity of extracts was analysed using the 2,2-diphenyl-1-picrylhydrazyl method. The effects of temperature, solvent type, and concentration on the phenolic extraction efficiency and antioxidant activity were studied using chemometric and statistical methods. The results showed that the extracts of V. album subsp. abietis contained large amounts of polyphenols and flavonoids (up to 57.673 mg g−1 and 9.955 mg g−1 of dry extract, respectively) and exhibited potent antioxidant activity, hence representing promising sources of powerful antioxidants. Due to its high extraction efficiency and considerable saving of time and solvent, ASE was more effective than the other extraction techniques. Extracts prepared with water-polar solvent mixtures displayed the highest TPC, TFC, and antioxidant activity, while organic polar solvents were the least efficient extractants.

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