Assessment of total bacterial cells in extended aeration activated sludge plants using flow cytometry as a microbial monitoring tool

2015 ◽  
Vol 22 (15) ◽  
pp. 11446-11455 ◽  
Author(s):  
Tarik Abzazou ◽  
Humbert Salvadó ◽  
Carmina Bruguera-Casamada ◽  
Pedro Simón ◽  
Carlos Lardín ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Activated Sludge ◽  
Bacterial Cells ◽  
Monitoring Tool ◽  
Microbial Monitoring

scholarly journals Using flow cytometry and light-induced fluorescence technique to characterizethe variability and characteristics of bioaerosols in springtime at Metro Atlanta, Georgia

2018 ◽  
Author(s):  
Arnaldo Negron ◽  
Natasha DeLeon-Rodriguez ◽  
Samantha M. Waters ◽  
Luke D. Ziemba ◽  
Bruce Anderson ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Nucleic Acid ◽  
Acid Content ◽  
Fungal Spores ◽  
Epifluorescence Microscopy ◽  
Nucleic Acid Content ◽  
Rain Event ◽  
Bacterial Cells ◽  
Sampling System ◽  
Spores And Pollen

Abstract. The abundance and speciation of primary biological aerosol particles (PBAP) is important for understanding their impacts on human health, cloud formation and ecosystems. Towards this, we have developed a protocol for quantifying PBAP collected from large volumes of air with a portable wet-walled cyclone bioaerosol sampler. A flow cytometry (FCM) protocol was then developed to quantify and characterize the PBAP populations from the sampler, which were confirmed against epifluorescence microscopy. The sampling system and FCM analysis were used to study PBAP in Atlanta, GA over a two-month period and showed clearly defined populations of DNA-containing particles: Low Nucleic Acid-content particles (bioLNA), High Nucleic Acid-content particles (HNA) being fungal spores and pollen. We find that daily-average springtime PBAP concentration (1 to 5 μm diameter) ranged between 1.4 × 104 and 1.1 × 105 m−3. The BioLNA population dominated PBAP during dry days (72 ± 18 %); HNA dominated the PBAP during humid days and following rain events, where HNA (e.g., wet-ejected fungal spores) comprised up to 92 % of the PBAP number. Concurrent measurements with a Wideband Integrated Bioaerosol Sensor (WIBS-4A) showed that FBAP and total FCM counts are similar; HNA (from FCM) significantly correlated with ABC type FBAP concentrations throughout the sampling period (and for the same particle size range, 1–5 μm diameter). However, the FCM bioLNA population, possibly containing bacterial cells, did not correlate to any FBAP type. The lack of correlation of any WIBS FBAP type with the bioLNA suggest bacterial cells may be more difficult to detect with autofluorescence than previously thought. Ιdentification of bacterial cells even in the FCM (bioLNA population) is challenging, given that the fluorescence level of stained cells at times may be comparable to that seen from abiotic particles. HNA and ABC displayed highest concentration on a humid and warm day after a rain event (4/14), suggesting that both populations correspond to wet-ejected fungal spores. Overall, information from both instruments combined reveals a highly dynamic airborne bioaerosol community over Atlanta, with a considerable presence of fungal spores during humid days, and a bioLNA population dominating bioaerosol community during dry days.

Preliminary Study on In Situ Realtime Quantitation of Target Bacteria on the Principle of Flow Cytometry

2017 ◽  
Vol 262 ◽  
pp. 224-227
Author(s):  
Gen Murakami ◽  
Yuichi Sugai ◽  
Kyuro Sasaki
Keyword(s):  
Flow Cytometry ◽  
Bacterial Culture ◽  
Scattered Light ◽  
Bacterial Suspension ◽  
Culture Solution ◽  
Bacterial Cells ◽  
Monitoring Wells ◽  
Measurement Principle ◽  
Preliminary Study

In-situ realtime method that can monitor the target bacteria should be used to determine the real situation of the bacteria in deep parts of heaps in heap bioleaching plants. This study suggest to apply flow cytometry technology to in-situ realtime monitoring of target bacteria. Flow cytometry is a method that can rapidly quantify the bacterial cells in bacterial suspension based on the detection of lights that are emitted from bacterial cells. In this study, we estimated the possibility of the application of flow cytometry to the selective detection of target bacteria. The bacterial culture solution that had been diluted by water including other bacteria was provided for fluorescence spectral analysis and scattered light analysis that were functions of flow cytometry. Our target bacteria could be selectively detected by those analyses in this study, therefore, it was shown that the flow cytometry could be useful for detecting target bacteria selectively. Because the measurement principle of flow cytometry is quite simple, it can be expected to be installed into deep heaps through the monitoring wells and determine the dominance of target bacteria in-situ and realtime in the future.

scholarly journals THE INFLUENCE OF ELECTROMAGNETIC FIELD ON MORPHOPHYSIOLOGICAL PARAMETERS OF BACTERIAL CELLS EVALUATED THROUGH FLOW CYTOMETRY

2016 ◽  
Author(s):  
Elena Radu ◽  
◽  
F. Marinescu ◽  
I. Savin ◽  
C. M. Kamerzan (Saviuc) ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Electromagnetic Field ◽  
Bacterial Cells ◽  
Through Flow

scholarly journals Probing the viability of palladium-challenged bacterial cells using flow cytometry

2018 ◽  
Vol 94 (1) ◽  
pp. 295-301 ◽  
Author(s):  
Jacob B Omajali ◽  
Iryna P Mikheenko ◽  
Tim W Overton ◽  
Mohamed L Merroun ◽  
Lynne E Macaskie
Keyword(s):  
Flow Cytometry ◽  
Bacterial Cells

Better understanding of the activated sludge process combining fluorescence-based methods and flow cytometry: A case study

2020 ◽  
Vol 90 ◽  
pp. 51-58 ◽  
Author(s):  
Vanesa Benito ◽  
Javier Etxebarria ◽  
Felipe Goñi-de-Cerio ◽  
Iñigo Gonzalez ◽  
Pilar Brettes ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Activated Sludge ◽  
Activated Sludge Process

scholarly journals A flow cytometry method for bacterial quantification and biomass estimates in activated sludge

2019 ◽  
Vol 160 ◽  
pp. 73-83 ◽  
Author(s):  
M.R. Brown ◽  
C.L. Hands ◽  
T. Coello-Garcia ◽  
B.S. Sani ◽  
A.I.G. Ott ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Activated Sludge ◽  
Bacterial Quantification

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.

scholarly journals Determination of Total Protein Content of Bacterial Cells by SYPRO Staining and Flow Cytometry

1999 ◽  
Vol 65 (7) ◽  
pp. 3251-3257 ◽  
Author(s):  
Mikhail V. Zubkov ◽  
Bernhard M. Fuchs ◽  
Heike Eilers ◽  
Peter H. Burkill ◽  
Rudolf Amann
Keyword(s):  
Flow Cytometry ◽  
Protein Content ◽  
Total Protein ◽  
Total Protein Content ◽  
Bacterial Cells ◽  
Protein Fluorescence ◽  
The North ◽  
Planktonic Bacteria ◽  
Different Temperatures ◽  
Protein Cell

ABSTRACT An assay has been developed for measuring protein biomass of marine planktonic bacteria by flow cytometry. The method was calibrated by using five species of Bacteria (an Arcobactersp., a Cytophaga sp., an Oceanospirillum sp., aPseudoalteromonas sp., and a Vibrio sp.) recently isolated from seawater samples and grown in culture at different temperatures. The intensity of SYPRO-protein fluorescence of these bacteria strongly correlated with their total protein content, measured by the bicinchoninic acid method to be in the range of 60 to 330 fg of protein cell−1(r 2 = 0.93, n = 34). According to the calibration, the mean biomass of planktonic bacteria from the North Sea in August 1998 was 24 fg of protein cell−1.

Protein extraction from activated sludge

2006 ◽  
Vol 54 (1) ◽  
pp. 175-181 ◽  
Author(s):  
M. Denecke
Keyword(s):  
Activated Sludge ◽  
Crude Extract ◽  
Extracellular Polymeric Substances ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Bacterial Cells ◽  
Uronic Acids ◽  
Isolated Cells ◽  
Humic Compounds ◽  
Immunoblot Technique

Two methods for the separation of protein originating from activated sludge were compared. In one method, the total protein was isolated out of the activated sludge (crude extract). These samples included all dissolved proteins originating from the bacterial cells and biofilm made up of extracellular polymeric substances (EPS). Every time polyacrylamide gel electrophoresis (PAGE) was done, the protein bands from samples of crude extract were covered by polymeric substances including carbohydrates, uronic acids or humic compounds. Using the immunoblot technique it was possible to demonstrate the presence of the heat shock protein HSP70 in crude extracts of activated sludge. The comparison of protein fingerprints required that clear and distinct bands appear on the PAGE analysis. To this end, a procedure to separates bacterial cells from the EPS was developed. Bacterial cells were separated by incubation with EDTA and subsequent filtration. The isolated cells were directly incubated in a sample buffer.

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