Fluorometric detection of phages in liquid media: Application to turbid samples

Rajnovic, Denis; Mas, Jordi

doi:10.1016/j.aca.2020.03.016

Cited by 7 publications

(2 citation statements)

References 40 publications

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“…For example, imaging flow cytometry was used to quantify somatic coliphages phiX174 and E. coli phage Qβ were detected with high specificity (within a concentration range of 10 4 -10 7 PFU/ml) providing results in 1 h with results comparable to the double-layer agar technique (Yang et al, 2019 ). Rajnovic and Mas ( 2020 ) evaluated the sensitivity of a resazurin-reporter sensor in E. coli DSMZ 613/T4 phage model using a fluorimeter. Viable bacterial host cells reduce resazurin to a fluorescent compound resorufin which can be enumerated and phage-induced changes of activity of the bacterial host are measured using resazurin as a reporter.…”

Section: Tools For Monitoring Phage and Viral Transience And Loadingmentioning

confidence: 99%

Coliphages as viral indicators of sanitary significance for drinking water

2022

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Coliphages are virus that infect coliform bacteria and are used in aquatic systems for risk assessment for human enteric viruses. This mini-review appraises the types and sources of coliphage and their fate and behavior in source waters and engineered drinking water treatment systems. Somatic (cell wall infection) and F+ (male specific) coliphages are abundant in drinking water sources and are used as indicators of fecal contamination. Coliphage abundances do not consistently correlate to human enteric virus abundance, but they suitably reflect the risks of exposure to human enteric viruses. Coliphages have highly variable surface characteristics with respect to morphology, size, charge, isoelectric point, and hydrophobicity which together interact to govern partitioning and removal characteristics during water treatment. The groups somatic and F+ coliphages are valuable for investigating the virus elimination during water treatment steps and as indicators for viral water quality assessment. Strain level analyses (e.g., Qβ or GA-like) provide more information about specific sources of viral pollution but are impractical for routine monitoring. Consistent links between rapid online monitoring tools (e.g., turbidity, particle counters, and flow cytometry) and phages in drinking water have yet to be established but are recommended as a future area of research activity. This could enable the real-time monitoring of virus and improve the process understanding during transient operational events. Exciting future prospects for the use of coliphages in aquatic microbiology are also discussed based on current scientific evidence and practical needs.

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Section: Tools For Monitoring Phage and Viral Transience And Loadingmentioning

confidence: 99%

Coliphages as viral indicators of sanitary significance for drinking water

2022

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“…Although several modifications have been made over the years (Cornax et al ., 1990; Lillehaug, 1997; Kropinski et al ., 2009; Cormier and Janes, 2014), the double agar layer method is still time‐consuming and labor intensive. In recent decades, polymerase chain reaction (PCR) (del Rio et al ., 2007; del Rio et al ., 2008; Ly‐Chatain et al ., 2011), Raman spectroscopy (Tayyarcan et al ., 2018), immunoassays (Larsson et al ., 2013; Khan et al ., 2015), resazurin reduction (Rajnovic and Mas, 2020) and mass spectrometry (Serafim et al ., 2017; Stverakova et al ., 2018) have been used to facilitate the detection of phages. Furthermore, flow cytometry (Michelsen et al ., 2007), fluorescence microscopy (Wang and Nitin, 2014), enzyme release (Stanek and Falkinham, 2001; Luna et al ., 2009; Harada et al ., 2018), surface plasmon resonance (García‐Aljaro et al ., 2008; Altintas et al ., 2015), impedance measurements (García‐Aljaro et al ., 2009) and optical density kinetics (Rajnovic et al ., 2019) have been employed to determine the lysis of host bacteria upon phage infection.…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of temperate bacteriophage using a simple motility assay

Guo

Lin

Wang

2021

Environ Microbiol Rep

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Summary Phage contamination is a common complication for the fermentation and pharmaceutical industries. The risk of bacteriophage contamination in laboratory processes increases with multiple rounds of genetic manipulation such as deletion and complementation. The contamination of temperate phages does not lead to immediate host cell lysis but could become a serious issue when the lytic cycle is activated under specific conditions. Our objective was to develop a quick and reliable detection method for checking possible temperate phage contamination. Here, using motility plates, we found that when the strain carries a newly acquired temperate phage, its presence can be easily detected by the formation of a clear ‘lysis zone’ when swimming against the original strain on the same swimming plates. Compared to the traditional double agar layer method and genomic sequencing‐based methods, the duration of the motility‐based assay is shorter and the procedure is simplified. More importantly, for the bacterial strains that already contain active prophages, this method can still easily detect the newly acquired phages without tedious phage identification procedure. These features make this method highly applicable to laboratory and industrial processes.

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