The utility of flow cytometry for potable reuse.

Protecting public health from pathogens is critical when treating wastewater to drinking water standards (i.e., planned water reuse). Viruses are a principal concern, yet real-time monitoring strategies do not currently measure virus removal through reuse processes. Flow cytometry (FCM) has enabled rapid and sensitive bacteria monitoring in water treatment applications, but methods for virus and protozoa monitoring remain immature. We discuss recent advances in the FCM field and FCM applications for quantifying microorganisms in water. We focus on flow virometry (FVM) developments, as virus enumeration methods show promise for water reuse applications. Ultimately, we propose FVM for near real-time monitoring across treatment to more accurately validate virus particle removal and for pilot studies to characterize removal through understudied unit processes.

[1]  H. Steen,et al.  Staining of Escherichia coli for flow cytometry: influx and efflux of ethidium bromide. , 1994, Cytometry.

[2]  Natasha S. Barteneva,et al.  Sorting of small infectious virus particles by flow virometry reveals distinct infectivity profiles , 2014, Nature Communications.

[3]  J. Tilton,et al.  High sensitivity detection and sorting of infectious human immunodeficiency virus (HIV-1) particles by flow virometry. , 2017, Virology.

[4]  W. D. de Vos,et al.  Genetic Diversity of Viable, Injured, and Dead Fecal Bacteria Assessed by Fluorescence-Activated Cell Sorting and 16S rRNA Gene Analysis , 2005, Applied and Environmental Microbiology.

[5]  P. Servais,et al.  Coupling Bacterial Activity Measurements with Cell Sorting by Flow Cytometry , 1999, Microbial Ecology.

[6]  P. Foladori,et al.  Assessment of activated sludge viability with flow cytometry. , 2002, Water research.

[7]  H. Salvadó,et al.  Assessment of total bacterial cells in extended aeration activated sludge plants using flow cytometry as a microbial monitoring tool , 2015, Environmental Science and Pollution Research.

[8]  M. Sinreich,et al.  Microbiological monitoring and classification of karst springs , 2013, Environmental Earth Sciences.

[9]  O. Köster,et al.  Flow-cytometric total bacterial cell counts as a descriptive microbiological parameter for drinking water treatment processes. , 2008, Water research.

[10]  L. Jones,et al.  Characterization of SYBR Gold nucleic acid gel stain: a dye optimized for use with 300-nm ultraviolet transilluminators. , 1999, Analytical biochemistry.

[11]  Thomas Egli,et al.  Rapid Detection and Enumeration of Giardia lamblia Cysts in Water Samples by Immunomagnetic Separation and Flow Cytometric Analysis , 2011, Applied and Environmental Microbiology.

[12]  W. Fitzgerald,et al.  Nanoparticle-based flow virometry for the analysis of individual virions. , 2013, The Journal of clinical investigation.

[13]  P. Forterre,et al.  Fake virus particles generated by fluorescence microscopy. , 2013, Trends in microbiology.

[14]  William K. W. Li,et al.  DNA distributions in planktonic bacteria stained with TOTO or TO‐PRO , 1995 .

[15]  Thammarat Koottatep,et al.  Comparison of rapid methods for detection of Giardia spp. and Cryptosporidium spp. (oo)cysts using transportable instrumentation in a field deployment. , 2012, Environmental science & technology.

[16]  J. Le Pecq Use of ethidium bromide for separation and determination of nucleic acids of various conformational forms and measurement of their associated enzymes. , 1971, Methods of biochemical analysis.

[17]  M. V. van Gemert,et al.  Single vs. swarm detection of microparticles and exosomes by flow cytometry , 2012, Journal of thrombosis and haemostasis : JTH.

[18]  J S Vrouwenvelder,et al.  Flow cytometric bacterial cell counts challenge conventional heterotrophic plate counts for routine microbiological drinking water monitoring. , 2017, Water research.

[19]  Yingying Wang,et al.  The impact of industrial-scale cartridge filtration on the native microbial communities from groundwater. , 2008, Water research.

[20]  Romaric Lacroix,et al.  Overcoming limitations of microparticle measurement by flow cytometry. , 2010, Seminars in thrombosis and hemostasis.

[21]  Sunny C. Jiang,et al.  Near Real-Time Flow Cytometry Monitoring of Bacterial and Viral Removal Efficiencies during Water Reclamation Processes , 2016 .

[22]  Yingying Wang,et al.  Rapid quantification of bacteria and viruses in influent, settled water, activated sludge and effluent from a wastewater treatment plant using flow cytometry. , 2013, Water science and technology : a journal of the International Association on Water Pollution Research.

[23]  D Marie,et al.  Application of the novel nucleic acid dyes YOYO-1, YO-PRO-1, and PicoGreen for flow cytometric analysis of marine prokaryotes , 1996, Applied and environmental microbiology.

[24]  C. Gerba,et al.  Viral Aggregation: Impact on Virus Behavior in the Environment. , 2017, Environmental science & technology.

[25]  F. Hammes,et al.  Identifying the underlying causes of biological instability in a full-scale drinking water supply system. , 2018, Water research.

[26]  Frederik Hammes,et al.  Short-term microbial dynamics in a drinking water plant treating groundwater with occasional high microbial loads. , 2016, Water research.

[27]  Lisa Zeigler Allen,et al.  Single Virus Genomics: A New Tool for Virus Discovery , 2011, PloS one.

[28]  D. Veal,et al.  Analysis‐only detection of Giardia by combining immunomagnetic separation and two‐color flow cytometry , 2003, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[29]  Rebecca A. Trenholm,et al.  Development of surrogate correlation models to predict trace organic contaminant oxidation and microbial inactivation during ozonation. , 2012, Water research.

[30]  D. Cliver,et al.  Pretreatment to avoid positive RT-PCR results with inactivated viruses. , 2002, Journal of virological methods.

[31]  Christopher Yu,et al.  Evaluation of methods for reverse osmosis membrane integrity monitoring for wastewater reuse , 2015 .

[32]  R. Lippé Flow Virometry: a Powerful Tool To Functionally Characterize Viruses , 2017, Journal of Virology.

[33]  R. Haugland,et al.  Characterization of PicoGreen reagent and development of a fluorescence-based solution assay for double-stranded DNA quantitation. , 1997, Analytical biochemistry.

[34]  T P Curtis,et al.  Flow cytometric quantification of viruses in activated sludge. , 2015, Water research.

[35]  O. Köster,et al.  Abundance and composition of indigenous bacterial communities in a multi-step biofiltration-based drinking water treatment plant. , 2014, Water research.

[36]  C. Brussaard,et al.  Virus production in phosphorus-limited Micromonas pusilla stimulated by a supply of naturally low concentrations of different phosphorus sources, far into the lytic cycle. , 2016, FEMS microbiology ecology.

[37]  Wayne L Chandler,et al.  Measurement of microvesicle levels in human blood using flow cytometry , 2016, Cytometry. Part B, Clinical cytometry.

[38]  Willy Verstraete,et al.  Past, present and future applications of flow cytometry in aquatic microbiology. , 2010, Trends in biotechnology.

[39]  R. W. Sabnis Handbook of Fluorescent Dyes and Probes , 2015 .

[40]  M C M van Loosdrecht,et al.  Monitoring microbiological changes in drinking water systems using a fast and reproducible flow cytometric method. , 2013, Water research.

[41]  N. Ashbolt,et al.  Application of flow cytometric methods for the routine detection of Cryptosporidium and Giardia in water. , 1994, Cytometry.

[42]  Randal J. Schoepp,et al.  Evaluation of ViroCyt® Virus Counter for Rapid Filovirus Quantitation , 2015, Viruses.

[43]  Trevor Douglas,et al.  Protein cage assembly across multiple length scales. , 2018, Chemical Society reviews.

[44]  Jurg Keller,et al.  Reverse osmosis integrity monitoring in water reuse: The challenge to verify virus removal - A review. , 2016, Water research.

[45]  B. Monger,et al.  Flow Cytometric Analysis of Marine Bacteria with Hoechst 33342 , 1993, Applied and environmental microbiology.

[46]  Greg Leslie,et al.  Removal Efficiency and Integrity Monitoring Techniques for Virus Removal by Membrane Processes , 2012 .

[47]  T G van Leeuwen,et al.  Optical and non‐optical methods for detection and characterization of microparticles and exosomes , 2010, Journal of thrombosis and haemostasis : JTH.

[48]  P. Lebaron,et al.  Comparison of Blue Nucleic Acid Dyes for Flow Cytometric Enumeration of Bacteria in Aquatic Systems , 1998, Applied and Environmental Microbiology.

[49]  Frederik Hammes,et al.  A pipeline for developing and testing staining protocols for flow cytometry, demonstrated with SYBR Green I and propidium iodide viability staining. , 2016, Journal of microbiological methods.

[50]  R. Lippé,et al.  Analysis of herpes simplex virus type I nuclear particles by flow cytometry , 2012, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[51]  Jannis Epting,et al.  Online flow cytometry reveals microbial dynamics influenced by concurrent natural and operational events in groundwater used for drinking water treatment , 2016, Scientific Reports.

[52]  Terry M. Gellner,et al.  Comparative effectiveness of membrane bioreactors, conventional secondary treatment, and chlorine and UV disinfection to remove microorganisms from municipal wastewaters. , 2012, Water research.

[53]  J Lizbeth Reyes Zamora,et al.  Flow virometry as a tool to study viruses. , 2017, Methods.

[54]  Bernhard Sonnleitner,et al.  Development and laboratory‐scale testing of a fully automated online flow cytometer for drinking water analysis , 2012, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[55]  C. Brussaard Optimization of Procedures for Counting Viruses by Flow Cytometry , 2004, Applied and Environmental Microbiology.

[56]  Feng Chen,et al.  Application of Digital Image Analysis and Flow Cytometry To Enumerate Marine Viruses Stained with SYBR Gold , 2001, Applied and Environmental Microbiology.

[57]  R. Lippé,et al.  Quantitative Evaluation of Protein Heterogeneity within Herpes Simplex Virus 1 Particles , 2017, Journal of Virology.

[58]  B. Pozzetto,et al.  Use of Flow Cytometry To Monitor Legionella Viability , 2008, Applied and Environmental Microbiology.

[59]  G. Bratbak,et al.  Enumeration of Marine Viruses in Culture and Natural Samples by Flow Cytometry , 1999, Applied and Environmental Microbiology.

[60]  P Foladori,et al.  Direct quantification of bacterial biomass in influent, effluent and activated sludge of wastewater treatment plants by using flow cytometry. , 2010, Water research.

[61]  Howard M. Shapiro,et al.  Practical Flow Cytometry , 1985 .

[62]  S. Sørensen,et al.  A low-cost, multiplexable, automated flow cytometry procedure for the characterization of microbial stress dynamics in bioreactors , 2013, Microbial Cell Factories.

[63]  C. Brussaard Quantification of aquatic viruses by flow cytometry , 2016 .