The development and use of real-time PCR for the quantification of nitrifiers in activated sludge

2002 ◽  
Vol 46 (1-2) ◽  
pp. 267-272 ◽  
Author(s):  
S.J. Hall ◽  
P. Hugenholtz ◽  
N. Siyambalapitiya ◽  
J. Keller ◽  
L.L. Blackall
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Real Time ◽  
Real Time Pcr ◽  
Wastewater Treatment Plants ◽  
Quantitative Polymerase Chain Reaction ◽  
Analytical Data ◽  
Full Scale ◽  
Rrna Gene ◽  
Dna Templates

Chemical analytical data has long been used to monitor the performance of activated sludge plants even though the process relies on the performance of microorganisms. It is now evident that a rapid and reliable quantitative method is required, to be able to monitor the organisms responsible for nutrient transformation and their activities, allowing avenues for more efficient nutrient removal. The development of real-time or quantitative polymerase chain reaction (PCR) also known as TaqMan® or 5′-nuclease assay has allowed the rapid, quantitative analysis of DNA templates, eliminating some of the variability traditionally associated with other quantitative techniques. In this study analysis of Nitrospira spp., one of the key organisms in nitrite oxidation in wastewater treatment, was used to validate real-time PCR for the their quantification in activated sludge. A probe and primer set, targeting the 16S rRNA gene of Nitrospira spp. was designed according to the constraints of the TaqMan® specifications. Samples used to evaluate the method included DNA from the sludge from full-scale wastewater treatment plants and laboratory scale systems. The reproducibility, quantitative efficiency and specificity were assessed in the evaluation. It was concluded that the method is sensitive and reproducible but has some constraints on the quantitative efficiency. A survey of full-scale systems for Nitrospira spp. was carried out and the results are presented here.

scholarly journals The capacity of wastewater treatment plants drives bacterial community structure and its assembly

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Young Kyung Kim ◽  
Keunje Yoo ◽  
Min Sung Kim ◽  
Il Han ◽  
Minjoo Lee ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Community Structure ◽  
Bacterial Community ◽  
Activated Sludge ◽  
Community Assembly ◽  
Bacterial Communities ◽  
Wastewater Treatment Plants ◽  
High Capacity ◽  
Operating Conditions ◽  
Rrna Gene

Abstract Bacterial communities in wastewater treatment plants (WWTPs) affect plant functionality through their role in the removal of pollutants from wastewater. Bacterial communities vary extensively based on plant operating conditions and influent characteristics. The capacity of WWTPs can also affect the bacterial community via variations in the organic or nutrient composition of the influent. Despite the importance considering capacity, the characteristics that control bacterial community assembly are largely unknown. In this study, we discovered that bacterial communities in WWTPs in Korea and Vietnam, which differ remarkably in capacity, exhibit unique structures and interactions that are governed mainly by the capacity of WWTPs. Bacterial communities were analysed using 16S rRNA gene sequencing and exhibited clear differences between the two regions, with these differences being most pronounced in activated sludge. We found that capacity contributed the most to bacterial interactions and community structure, whereas other factors had less impact. Co-occurrence network analysis showed that microorganisms from high-capacity WWTPs are more interrelated than those from low-capacity WWTPs, which corresponds to the tighter clustering of bacterial communities in Korea. These results will contribute to the understanding of bacterial community assembly in activated sludge processing.

An investigation into studying of the activated sludge foaming potential by using physicochemical parameters

2002 ◽  
Vol 46 (1-2) ◽  
pp. 525-528 ◽  
Author(s):  
K. Hladikova ◽  
I. Ruzickova ◽  
P. Klucova ◽  
J. Wanner
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Physicochemical Parameters ◽  
Wastewater Treatment Plants ◽  
Physicochemical Characteristics ◽  
Full Scale ◽  
Aeration Tank ◽  
Foam Formation ◽  
Individual Parameter

This paper examines how the physicochemical characteristics of the solids are related to foam formation and describes how the foaming potential of full-scale plants can be assessed. The relations among activated sludge and biological foam hydrophobicity, scum index, aeration tank cover and filamentous population are evaluated. Individual parameter comparison reveals the scumming intensity can be estimated only on the assumption that foams is already established. None of the above mentioned characteristics can be reliably used to predict the foaming episodes at wastewater treatment plants.

scholarly journals The Microbial Database for Danish wastewater treatment plants with nutrient removal (MiDas-DK) – a tool for understanding activated sludge population dynamics and community stability

2013 ◽  
Vol 67 (11) ◽  
pp. 2519-2526 ◽  
Author(s):  
A. T. Mielczarek ◽  
A. M. Saunders ◽  
P. Larsen ◽  
M. Albertsen ◽  
M. Stevenson ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Nutrient Removal ◽  
Wastewater Treatment Plants ◽  
Amplicon Sequencing ◽  
Temporal Variations ◽  
Full Scale ◽  
16S Rrna Amplicon Sequencing ◽  
Specific Factors ◽  
Operational Data

Since 2006 more than 50 Danish full-scale wastewater treatment plants with nutrient removal have been investigated in a project called ‘The Microbial Database for Danish Activated Sludge Wastewater Treatment Plants with Nutrient Removal (MiDas-DK)’. Comprehensive sets of samples have been collected, analyzed and associated with extensive operational data from the plants. The community composition was analyzed by quantitative fluorescence in situ hybridization (FISH) supported by 16S rRNA amplicon sequencing and deep metagenomics. MiDas-DK has been a powerful tool to study the complex activated sludge ecosystems, and, besides many scientific articles on fundamental issues on mixed communities encompassing nitrifiers, denitrifiers, bacteria involved in P-removal, hydrolysis, fermentation, and foaming, the project has provided results that can be used to optimize the operation of full-scale plants and carry out trouble-shooting. A core microbial community has been defined comprising the majority of microorganisms present in the plants. Time series have been established, providing an overview of temporal variations in the different plants. Interestingly, although most microorganisms were present in all plants, there seemed to be plant-specific factors that controlled the population composition thereby keeping it unique in each plant over time. Statistical analyses of FISH and operational data revealed some correlations, but less than expected. MiDas-DK (www.midasdk.dk) will continue over the next years and we hope the approach can inspire others to make similar projects in other parts of the world to get a more comprehensive understanding of microbial communities in wastewater engineering.

scholarly journals Quantification of biologically and chemically bound phosphorus in activated sludge from full-scale plants with biological P-removal

2021 ◽  
Author(s):  
Francesca Petriglieri ◽  
Jette F. Petersen ◽  
Miriam Peces ◽  
Marta Nierychlo ◽  
Kamilla Hansen ◽  
...  
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Mass Balance ◽  
Wastewater Treatment Plants ◽  
Raman Microspectroscopy ◽  
Full Scale ◽  
P Fractions ◽  
P Content ◽  
Total P ◽  
P Removal

AbstractLarge amounts of phosphorus (P) are present in activated sludge from municipal wastewater treatment plants, where it exists in the form of metal salt precipitates or biologically bound into the biomass as nucleic acids, cell membrane components, and the extracellular polymeric substances or, in special polyphosphate-accumulating organisms (PAOs), as intracellular polyphosphate. Only recently, methods that reliably allow an absolute quantification of the different P-fractions, such as sequential extraction, Raman microspectroscopy, solid-state 31P magic angle spinning (MAS) NMR, and solution state 31P NMR have been developed. This study combines these techniques to obtain a comprehensive P mass-balance of activated sludge from four wastewater treatment plants with enhanced biological phosphate removal (EBPR). The total content of P and various cations was measured by chemical analysis (ICP-OES), and different P fractions were extracted for chemical characterization. Chemically bound P constituted 38-69% of total P, most likely in the form of Fe, Mg, or Al minerals, while organically bound P constituted 7-9%. By using Raman microspectroscopy and solution state 31P NMR and 31P MAS NMR spectroscopy before and after anaerobic P-release experiments, poly-P was quantified and constituted 22-54% of total P in the activated sludges and was found in approx. 25% of all bacterial cells. Moreover, Raman microspectroscopy in combination with fluorescence in situ hybridization (FISH) was used to quantify the species-specific intracellular poly-P of known PAO genera (Tetrasphaera, Ca. Accumulibacter, Dechloromonas) and other microorganisms known to possess high level of poly-P, such as the filamentous Ca. Microthrix. They were all abundant, as measured by quantitative-FISH and amplicon sequencing, and accumulated large amount of poly-P, depending on their cell-size, contributing substantially to the P-removal. Interestingly, in all four EBPR plants investigated, only 1-13% of total poly-P was stored by unidentified PAO, highlighting that most PAOs in the full-scale EBPR plants investigated are now known.HighlightsExhaustive P mass-balance of main organic and inorganic P-species in four EBPR plantsQuantification of poly-P of FISH-defined PAO and other species with high P contentTotal P content was 36-50 mgP/gSS of which 31-62% was in biomass and as poly-PA high fraction of all cells (25-30%) contained a high content of poly-PKnown PAOs contained almost all poly-P in the EBPR plants investigated

Abundance of amoA genes of ammonia-oxidizing archaea and bacteria in activated sludge of full-scale wastewater treatment plants

2011 ◽  
Vol 102 (4) ◽  
pp. 3694-3701 ◽  
Author(s):  
Tawan Limpiyakorn ◽  
Puntipar Sonthiphand ◽  
Chaiwat Rongsayamanont ◽  
Chongrak Polprasert
Keyword(s):  
Wastewater Treatment ◽  
Activated Sludge ◽  
Wastewater Treatment Plants ◽  
Full Scale ◽  
Ammonia Oxidizing Archaea

Enumeration of viable Escherichia coli by real-time PCR with propidium monoazide

2012 ◽  
Vol 66 (10) ◽  
pp. 2065-2073 ◽  
Author(s):  
N. Yokomachi ◽  
J. Yaguchi
Keyword(s):  
Escherichia Coli ◽  
Wastewater Treatment ◽  
Real Time ◽  
Real Time Pcr ◽  
Wastewater Treatment Plants ◽  
Propidium Monoazide ◽  
E Coli ◽  
Cell Counts ◽  
Viable Cells ◽  
Heat Treated

A photo-inducible DNA-binding dye, propidium monoazide (PMA), was used to distinguish viable and dead Escherichia coli cells. Microscopic observations using a combination of the dyes 4′,6-diamidino-2-phenylindole and PMA indicated that PMA stained only dead cells, with membrane damage, red. Mixtures of viable and heat-treated E. coli cells were subjected to real-time polymerase chain reaction (PCR) with PMA treatment. Viable cell counts were linearly related to real-time PCR threshold cycle values for PMA-treated cells in the mixtures of viable and heat-treated cells, as long as the ratio of dead cells to viable cells was no greater than 10. In the wastewater treatment plants, total, viable and culturable E. coli were enumerated by real-time PCR, real-time PCR coupled with PMA treatment and the most probable number method using EC-MUG medium, respectively. The concentrations of viable E. coli in the wastewater treatment plants were much higher than those of culturable cells. In addition, viable cells were even more chlorine resistant than culturable ones.

Quantification of ammonia-oxidizing bacteria populations in full-scale sewage activated sludge systems and assessment of system variables affecting their performance

2006 ◽  
Vol 54 (1) ◽  
pp. 91-99 ◽  
Author(s):  
T. Limpiyakorn ◽  
F. Kurisu ◽  
O. Yagi
Keyword(s):  
Activated Sludge ◽  
Real Time ◽  
Real Time Pcr ◽  
Full Scale ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Ammonia Oxidizing Bacteria ◽  
Primer Sets ◽  
Activated Sludge Systems ◽  
Pcr Primer

This study carried out quantification of ammonia-oxidizing bacteria (AOB) populations in 12 full-scale sewage activated sludge systems that were different in ammonia removals and treatment processes during three different seasons. Experiment was divided into 3 parts: 1) analysis of AOB communities by PCR-DGGE-cloning-sequencing of 16S rRNA genes; 2) development of four real-time PCR primer sets for quantification of the particular AOB of interest; and 3) quantification of AOB populations by using the newly developed real-time PCR primer sets. The results suggested that all the primer sets gave good reproducibility and specificity for PCR amplification with the detection limits of 102 copies/PCR reaction. Although the 12 systems were different in several aspects, one of the identified sequence types of Nitrosomonas oligotropha cluster was the dominant AOB in every system and every season studied. However, the other sequence type of this cluster was not significantly involved in ammonia removals in the systems. The occurrence of N. communis cluster in the systems seemed to depend on the remaining oxygen concentrations in the sludge floc and thus the activity of aerobic heterotrophs in the aeration tanks. N. europaea–Nitrosococcus. mobilis solely existed in one A2O system of which the influent contained twice the chloride concentrations than those of other systems.

Modelling of wastewater treatment plants – how far shall we go with sophisticated modelling tools?

2006 ◽  
Vol 53 (3) ◽  
pp. 79-89 ◽  
Author(s):  
G.C. Glover ◽  
C. Printemps ◽  
K. Essemiani ◽  
J. Meinhold
Keyword(s):  
Fluid Dynamics ◽  
Computational Fluid Dynamics ◽  
Wastewater Treatment ◽  
Activated Sludge ◽  
Wastewater Treatment Plants ◽  
Pilot Scale ◽  
Full Scale ◽  
Structure Identification ◽  
Oxidation Ditch ◽  
Computational Fluid Dynamics Analysis

Several levels of complexity are available for modelling of wastewater treatment plants. Modelling local effects rely on computational fluid dynamics (CFD) approaches whereas activated sludge models (ASM) represent the global methodology. By applying both modelling approaches to pilot plant and full scale systems, this paper evaluates the value of each method and especially their potential combination. Model structure identification for ASM is discussed based on a full-scale closed loop oxidation ditch modelling. It is illustrated how and for what circumstances information obtained via CFD (computational fluid dynamics) analysis, residence time distribution (RTD) and other experimental means can be used. Furthermore, CFD analysis of the multiphase flow mechanisms is employed to obtain a correct description of the oxygenation capacity of the system studied, including an easy implementation of this information in the classical ASM modelling (e.g. oxygen transfer). The combination of CFD and activated sludge modelling of wastewater treatment processes is applied to three reactor configurations, a perfectly mixed reactor, a pilot scale activated sludge basin (ASB) and a real scale ASB. The application of the biological models to the CFD model is validated against experimentation for the pilot scale ASB and against a classical global ASM model response. A first step in the evaluation of the potential of the combined CFD-ASM model is performed using a full scale oxidation ditch system as testing scenario.

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