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  • 1
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    Data_Sheet_1.docx
    Frontiers Media SA ; 2022
    In:  Frontiers in Microbiology Vol. 13 ( 2022-5-20)
    Details
    In: Frontiers in Microbiology, Frontiers Media SA, Vol. 13 ( 2022-5-20)
    Kurzfassung: RNA virus populations are not clonal; rather, they comprise a mutant swarm in which sequences are slightly different from the master sequence. Genetic diversity within a population (intrapopulation genetic diversity) is critical for RNA viruses to survive under environmental stresses. Disinfection has become an important practice in the control of pathogenic viruses; however, the impact of intrapopulation genetic diversity on the sensitivity of disinfection, defined as –log 10 (postdisinfected infectious titer/predisinfected titer), has not been elucidated. In this study, we serially passaged populations of rhesus rotavirus. We demonstrated that populations with reduced chlorine sensitivity emerged at random and independently of chlorine exposure. Sequencing analysis revealed that compared with sensitive populations, less-sensitive ones had higher non-synonymous genetic diversity of the outer capsid protein gene, suggesting that changes in the amino acid sequences of the outer capsid protein were the main factors influencing chlorine sensitivity. No common mutations were found among less-sensitive populations, indicating that rather than specific mutations, the diversity of the outer capsid protein itself was associated with the disinfection sensitivity and that the disinfection sensitivity changed stochastically. Simulation results suggest that the disinfection sensitivity of a genetically diverse population is destabilized if cooperative viral clusters including multiple sequences are formed. These results advocate that any prevention measures leading to low intrapopulation genetic diversity are important to prevent the spread and evolution of pathogenic RNA viruses in society.
    Materialart: Online-Ressource
    ISSN: 1664-302X
    URL: Article
    URL: Article
    DOI: 10.3389/fmicb.2022.839513
    DOI: 10.3389/fmicb.2022.839513.s001
    Sprache: Unbekannt
    Verlag: Frontiers Media SA
    Publikationsdatum: 2022
    ZDB Id: 2587354-4
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  • 2
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    Bottleneck Size-Dependent Changes in the Genetic Diversity and Specific Growth Rate of a Rotavirus A Strain
    American Society for Microbiology ; 2020
    In:  Journal of Virology Vol. 94, No. 10 ( 2020-05-04)
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 94, No. 10 ( 2020-05-04)
    Kurzfassung: RNA viruses form a dynamic distribution of mutant swarms (termed “quasispecies”) due to the accumulation of mutations in the viral genome. The genetic diversity of a viral population is affected by several factors, including a bottleneck effect. Human-to-human transmission exemplifies a bottleneck effect, in that only part of a viral population can reach the next susceptible hosts. In the present study, two lineages of the rhesus rotavirus (RRV) strain of rotavirus A were serially passaged five times at a multiplicity of infection (MOI) of 0.1 or 0.001, and three phenotypes (infectious titer, cell binding ability, and specific growth rate) were used to evaluate the impact of a bottleneck effect on the RRV population. The specific growth rate values of lineages passaged under the stronger bottleneck (MOI of 0.001) were higher after five passages. The nucleotide diversity also increased, which indicated that the mutant swarms of the lineages under the stronger bottleneck effect were expanded through the serial passages. The random distribution of synonymous and nonsynonymous substitutions on rotavirus genome segments indicated that almost all mutations were selectively neutral. Simple simulations revealed that the presence of minor mutants could influence the specific growth rate of a population in a mutant frequency-dependent manner. These results indicate a stronger bottleneck effect can create more sequence spaces for minor sequences. IMPORTANCE In this study, we investigated a bottleneck effect on an RRV population that may drastically affect the viral population structure. RRV populations were serially passaged under two levels of a bottleneck effect, which exemplified human-to-human transmission. As a result, the genetic diversity and specific growth rate of RRV populations increased under the stronger bottleneck effect, which implied that a bottleneck created a new space in a population for minor mutants originally existing in a hidden layer, which includes minor mutations that cannot be distinguished from a sequencing error. The results of this study suggest that the genetic drift caused by a bottleneck in human-to-human transmission explains the random appearance of new genetic lineages causing viral outbreaks, which can be expected according to molecular epidemiology using next-generation sequencing in which the viral genetic diversity within a viral population is investigated.
    Materialart: Online-Ressource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/JVI.02083-19
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 2020
    ZDB Id: 1495529-5
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  • 3
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    Culture-Independent Evaluation of Nonenveloped-Virus Infectivity Reduced by Free-Chlorine Disinfection
    American Society for Microbiology ; 2015
    In:  Applied and Environmental Microbiology Vol. 81, No. 8 ( 2015-04-15), p. 2819-2826
    Details
    In: Applied and Environmental Microbiology, American Society for Microbiology, Vol. 81, No. 8 ( 2015-04-15), p. 2819-2826
    Kurzfassung: The inability of molecular detection methods to distinguish disinfected virions from infectious ones has hampered the assessment of infectivity for enteric viruses caused by disinfection practices. In the present study, the reduction of infectivity of murine norovirus S7-PP3 and mengovirus vMC0, surrogates of human noroviruses and enteroviruses, respectively, caused by free-chlorine treatment was characterized culture independently by detecting carbonyl groups on viral capsid protein. The amount of carbonyls on viral capsid protein was evaluated by the proportion of biotinylated virions trapped by avidin-immobilized gel (percent adsorbed). This culture-independent approach demonstrated that the percent adsorbed was significantly correlated with the logarithm of the infectious titer of tested viruses. Taken together with the results of previous reports, the result obtained in this study indicates that the amount of carbonyls on viral capsid protein of four important families of waterborne pathogenic viruses, Astroviridae , Reoviridae , Caliciviridae , and Picornaviridae , is increased in proportion to the received oxidative stress of free chlorine. There was also a significant correlation between the percent adsorbed and the logarithm of the ratio of genome copy number to PFU, which enables estimation of the infectious titer of a subject virus by measuring values of the total genome copy number and the percent adsorbed. The proposed method is applicable when the validation of a 4-log reduction of viruses, a requirement in U.S. EPA guidelines for virus removal from water, is needed along with clear evidence of the oxidation of virus particles with chlorine-based disinfectants.
    Materialart: Online-Ressource
    ISSN: 0099-2240 , 1098-5336
    URL: Article
    DOI: 10.1128/AEM.03802-14
    RVK:
    WA 15000
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 2015
    ZDB Id: 223011-2
    ZDB Id: 1478346-0
    SSG: 12
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  • 4
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    A new approach for evaluating the infectivity of noncultivatable enteric viruses without cell culture
    IWA Publishing ; 2013
    In:  Water Science and Technology Vol. 67, No. 10 ( 2013-05-01), p. 2236-2240
    Details
    In: Water Science and Technology, IWA Publishing, Vol. 67, No. 10 ( 2013-05-01), p. 2236-2240
    Kurzfassung: This study developed a novel approach for evaluating the infectivity of enteric viruses without cell culture. Cumulative carbonyl groups on the viral capsid protein were labeled using biotin hydrazide, and the biotinylated virions were separated using a spin column filled with avidin-immobilized gel. Rotavirus was treated with free chlorine at an initial concentration of 0.3 mg/L for 3 min, and the log reduction in the infectious titer was 0.19 log (standard deviation, SD = 0.05). The log reduction of rotavirus treated with free chlorine at an initial concentration of 0.6 mg/L for 3 min was 2.6 log (SD = 0.37). No significant reductions in the amplicon copy numbers were observed in these free chlorine-treated samples. The recovery levels of intact virions in the first three fractions after biotin-avidin affinity chromatography were 76, 21, and 2.8%, while those of virions treated with free chlorine at an initial concentration of 0.3 mg/L for 3 min were 70, 23, and 5.6%. These results showed that the proposed approach could discriminate a 0.19 log infectivity-reduced population from an intact population, although no reduction in the amplicon copy number was observed. This novel method could be applied to noncultivatable enteric viruses such as human norovirus and sapovirus, and it could be very helpful for evaluating the viral inactivation efficiencies of intervention measures.
    Materialart: Online-Ressource
    ISSN: 0273-1223 , 1996-9732
    URL: Article
    DOI: 10.2166/wst.2013.114
    Sprache: Englisch
    Verlag: IWA Publishing
    Publikationsdatum: 2013
    ZDB Id: 764273-8
    ZDB Id: 2024780-1
    SSG: 14
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  • 5
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    Bacterial histo-blood group antigens contributing to genotype-dependent removal of human noroviruses with a microfiltration membrane
    Elsevier BV ; 2016
    In:  Water Research Vol. 95 ( 2016-05), p. 383-391
    Details
    In: Water Research, Elsevier BV, Vol. 95 ( 2016-05), p. 383-391
    Materialart: Online-Ressource
    ISSN: 0043-1354
    URL: Article
    DOI: 10.1016/j.watres.2016.04.018
    Sprache: Englisch
    Verlag: Elsevier BV
    Publikationsdatum: 2016
    ZDB Id: 202613-2
    ZDB Id: 1501098-3
    SSG: 14
    Standort Signatur Einschränkungen Verfügbarkeit
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  • 6
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    Free-Chlorine Disinfection as a Selection Pressure on Norovirus
    American Society for Microbiology ; 2018
    In:  Applied and Environmental Microbiology Vol. 84, No. 13 ( 2018-07)
    Details
    In: Applied and Environmental Microbiology, American Society for Microbiology, Vol. 84, No. 13 ( 2018-07)
    Kurzfassung: Human noroviruses are excreted in feces from infected individuals and included in wastewater. It is critical to remove/inactivate them in wastewater treatment processes, particularly in the disinfection step, before release to aquatic environments. However, the high mutation rates of human noroviruses raise concerns about the emergence of strains that are less susceptible to disinfectants and can survive even after wastewater treatment. This study aimed to demonstrate the strain-dependent susceptibility of norovirus to free chlorine. A population originated from the murine norovirus strain S7-PP3, a surrogate for human noroviruses in environmental testing, was exposed to free chlorine and then propagated in a host cell. This cycle of free chlorine exposure followed by propagation in cells was repeated 10 times, and populations with lower susceptibility to free chlorine were obtained from two independent trials of chlorine exposure cycles. Open reading frame 2 (ORF2) and ORF3 of the murine norovirus genome were analyzed by next-generation sequencing, and a unique nonsynonymous mutation (corresponding to a change from phenylalanine to serine) at nucleotide (nt) 7280 in ORF3, which encodes the minor capsid protein VP2, was found in chlorine-exposed populations from both trials. It was confirmed that all of the clones from the chlorine-treated population had lower susceptibility to free chlorine than those from the control population. These results indicate that exposure to free chlorine and dilution exert different driving forces to form murine norovirus (MNV) quasispecies, and that there is a selective force to form MNV quasispecies under free chlorine exposure. IMPORTANCE This study showed that free chlorine disinfection exerted a selection pressure for murine norovirus (MNV). The strain-dependent viral susceptibility to the disinfectant elucidated in this study highlights the importance of employing less susceptible strains as representative viruses in disinfection tests, because the disinfection rate values obtained from more susceptible strains would be less useful in predicting the virus inactivation efficiency of circulating strains under practical disinfection conditions.
    Materialart: Online-Ressource
    ISSN: 0099-2240 , 1098-5336
    URL: Article
    DOI: 10.1128/AEM.00244-18
    RVK:
    WA 15000
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 2018
    ZDB Id: 223011-2
    ZDB Id: 1478346-0
    SSG: 12
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  • 7
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    Histo-Blood Group Antigen-Like Substances of Human Enteric Bacteria as Specific Adsorbents for Human Noroviruses
    American Society for Microbiology ; 2013
    In:  Journal of Virology Vol. 87, No. 17 ( 2013-09), p. 9441-9451
    Details
    In: Journal of Virology, American Society for Microbiology, Vol. 87, No. 17 ( 2013-09), p. 9441-9451
    Kurzfassung: Histo-blood group antigens (HBGAs) have been suggested to be receptors or coreceptors for human noroviruses (HuNoVs) expressed on the intestinal epithelium. We isolated an enteric bacterium strain (SENG-6), closely related to Enterobacter cloacae , bearing HBGA-like substances from a fecal sample of a healthy individual by using a biopanning technique with anti-HBGA antibodies. The binding capacities of four genotypes of norovirus-like particles (NoVLPs) to Enterobacter sp. SENG-6 cells were confirmed by enzyme-linked immunosorbent assay (ELISA). Transmission electron microscopy demonstrated that NoVLPs bound mainly to extracellular polymeric substances (EPS) of Enterobacter sp. SENG-6, where the HBGA-like substances were localized. EPS that contained HBGA-like substances extracted from Enterobacter sp. SENG-6 was shown by enzyme-linked immunosorbent assay (ELISA) to be capable of binding to NoVLPs of a GI.1 wild-type strain (8fIIa) and a GII.6 strain that can recognize A antigen but not to an NoVLP GI.1 mutant strain (W375A) that loses the ability to bind to A antigen. Enzymatic cleavage of terminal N -acetyl-galactosamine residues in the bacterial EPS weakened bacterial EPS binding to the GI.1 wild-type strain (8fIIa). These results indicate that A-like substances in the bacterial EPS play a key role in binding to NoVLPs. Since the specific binding of HuNoVs to HBGA-positive enteric bacteria is likely to affect the transmission and infection processes of HuNoVs in their hosts and in the environment, further studies of human enteric bacteria and their binding capacity to HuNoVs will provide a new scientific platform for understanding interactions between two types of microbes that were previously regarded as biologically unrelated.
    Materialart: Online-Ressource
    ISSN: 0022-538X , 1098-5514
    URL: Article
    DOI: 10.1128/JVI.01060-13
    Sprache: Englisch
    Verlag: American Society for Microbiology
    Publikationsdatum: 2013
    ZDB Id: 1495529-5
    Standort Signatur Einschränkungen Verfügbarkeit
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