Effects of Ionic Strength on Bacteriophage MS2 Behavior and Their Implications for the Assessment of Virus Retention by Ultrafiltration Membranes

Furiga, Aurélie; Pierre, Gwenaëlle; Glories, Marie; Aimar, Pierre; Roques, Christine; Causserand, Christel; Bergé, Mathieu

doi:10.1128/aem.01075-10

Cited by 42 publications

(22 citation statements)

References 40 publications

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“…The qRT-PCR was performed by using the SuperScript III Platinum SYBR Green one-step qRT-PCR kit (Life Technologies; Grand Island, NY) following the manufacturer's protocol. The primers and reaction parameters used for this one-step process were described in previous studies (Ogorzaly and Gantzer 2007;Furiga et al 2011). A 10-fold serial dilution of the commercial MS2 genomic RNA was used as a standard curve ranging from 4 to 4000 fg (2 £ 10 3 to 2 £ 10 6 genomes) in a 25 ml reaction.…”

Section: Ms2 Genomic Rna Extraction and Quantificationmentioning

confidence: 99%

Generation and sampling of nanoscale infectious viral aerosols

Walls

Ensor

Harvey

et al. 2016

Aerosol Science and Technology

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Airborne viruses represent a potentially significant health threat. However, only recently have researchers begun to characterize the size and infectivity of viral bioaerosols in the nanoscale size range. There are limitations in the generation of test viral aerosols and the ability to sample with acceptable efficiency. Reported here is use of a laminar-flow water condensation method to efficiently sample nanoscale bioaerosols to sizes well below 100 nm. We used MS2 bacteriophage in water to provide an aerosol with particles sizes from 300 nm down to 45 nm for sampling by both an all-glass impinger (4 mm; AGI-4) and the water condensation bioaerosol sampler. We demonstrated the existence of infectious viral particles below 100 nm and a higher collection efficiency by the water condensation sampler compared to the AGI-4 at nanoscale sizes. For example, the water condensation bioaerosol sampler that collected particles at 45 nm in diameter had 20 times more infective virions per collected particle compared to the AGI-4. However, when we corrected the AGI-4 data for particle size-dependent collection efficiency, the results were similar. We also used quantitative reverse transcription polymerase chain reaction, along with culturing for infectivity to determine the percent infectivity of the aerosol by particle size. Finally, we used a simple calculation to determine that a large fraction of sub-100 nm particles did not contain infectious virus because of the low titer concentration of virus in the Collison fluid.

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Section: Ms2 Genomic Rna Extraction and Quantificationmentioning

confidence: 99%

Generation and sampling of nanoscale infectious viral aerosols

Walls

Ensor

Harvey

et al. 2016

Aerosol Science and Technology

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“…To achieve this goal, Leibo and Mazur used harsh osmotic shock with 3 M NaCl in order to trigger DNA release and removal of virions from liquid media [ 12 ]. Phage filtration through induction of virion aggregation with salt was also addressed in the work of Furiga et al in 2011 [ 13 ]. In their study, aggregation of phage (RNA phage M2) was recognized as a strong inactivating factor, which dramatically reduced infectivity of phage.…”

Section: Introductionmentioning

confidence: 99%

Aggregation/dispersion transitions of T4 phage triggered by environmental ion availability

et al. 2017

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BackgroundBacteriophage survives in at least two extremes of ionic environments: bacterial host (high ionic-cytosol) and that of soil (low ionic-environmental water). The impact of ionic composition in the micro- and macro-environments has not so far been addressed in phage biology.ResultsHere, we discovered a novel mechanism of aggregation/disaggregation transitions by phage virions. When normal sodium levels in phage media (150 mM) were lowered to 10 mM, advanced imaging by scanning electron microscopy, atomic force microscopy and dynamic light scattering all revealed formation of viral packages, each containing 20–100 virions. When ionic strength was returned from low to high, the aggregated state of phage reversed to a dispersed state, and the change in ionic strength did not substantially affect infectivity of the phage. By providing the direct evidence, that lowering of the sodium ion below the threshold of 20 mM causes rapid aggregation of phage while returning Na+ concentration to the values above this threshold causes dispersion of phage, we identified a biophysical mechanism of phage aggregation.ConclusionsOur results implicate operation of group behavior in phage and suggest a new kind of quorum sensing among its virions that is mediated by ions. Loss of ionic strength may act as a trigger in an evolutionary mechanism to improve the survival of bacteriophage by stimulating aggregation of phage when outside a bacterial host. Reversal of phage aggregation is also a promising breakthrough in biotechnological applications, since we demonstrated here the ability to retain viable virion aggregates on standard micro-filters.

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“…While studying the effect of temperature on adsorption it was revealed that maximum percentage or nearing 100% adsorption was at 37 The presence of electrolytes in the phage-bacteria growth medium has a profound effect on the adsorption of phage to host cell. The ionic strength of the solution in which the phage is suspended induces significant effects on its behaviour (Adams, 1959;Furiga et al, 2011). 1M NaCl was found to be optimum for viability and 0.25M NaCl for adsorption.…”

Section: Discussionmentioning

confidence: 99%

Physico Chemical Characterization of a T5-Like Salmonella Phage Φsp-3

Augustine¹,

Bhat²

2014

J microb biotech food sci

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Antimicrobial resistance is a problem faced by humanity for quite some time now. Search for an intelligent alterative to antibiotics led the scientific community to join hands with an age old foe of pathogenic bacteria - bacteriophages. The present study elaborates the potential of a previously isolated phage with biocontrol capability, to survive and adsorb in various physicochemical environments . ΦSP-3, a Salmonella specific lytic phage capable of infecting host in nutrient deprived states with an added advantage of high host specificity and absence of virulence genes as previously reported was chosen for the study. Salmonella Enteritidis was used as host. Basic growth parameters like time taken for phage adsorption, optimal multiplicity of infection (MOI) and one step growth curve, were determined, followed by physicochemical characterization. Various parameters studied include temperature, pH, salinity and presence of sugars and CaCl2. 25 minutes of exposure time was required for 100% adsorption and optimal MOI was calculated as one. The latent period and the rise period was 30 minutes each with a subsequent burst size of 60 phages per bacterium. ΦSP-3 could survive upto 70ºC. 1M NaCl was optimal for phage viability while 0.25M favored maximal adsorption. The optimum pH for ΦSP-3 viability and adsorption was 8. Among the sugars,Arabinose was most influential in inactivating ΦSP-3. 100% adsorption was achieved at both 37ºC and at 40ºC. 10mM of CaCl2 to be optimum for ΦSP-3. Thus the present study substantiates the candidature of ΦSP-3 as a sturdy biocontrol agent capable of thriving well in diverse physicochemical environments.

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Effects of Ionic Strength on Bacteriophage MS2 Behavior and Their Implications for the Assessment of Virus Retention by Ultrafiltration Membranes

Cited by 42 publications

References 40 publications

Generation and sampling of nanoscale infectious viral aerosols

Generation and sampling of nanoscale infectious viral aerosols

Aggregation/dispersion transitions of T4 phage triggered by environmental ion availability

Physico Chemical Characterization of a T5-Like Salmonella Phage Φsp-3

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