Suitability of pepper mild mottle virus as a human enteric virus surrogate for assessing the efficacy of thermal or free-chlorine disinfection processes by using infectivity assays and enhanced viability PCR

Shirasaki, Nobutaka; Matsushita, Taku; Matsui, Yoshihiko; Koriki, S.

doi:10.1016/j.watres.2020.116409

Cited by 36 publications

(20 citation statements)

References 34 publications

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“…The inactivation rate constants by free chlorine ( k FC ) ranged from 3.9 to 13.4 mg –1 min –1 L. The predicted CT values for 4-log inactivation ranged from 0.69 to 2.36 mg min L –1 . The result of the predicted CT values for 4-log inactivation of CVB5 Faulkner (1.02 mg min L –1 ) was consistent with a previous study, reporting 1.15–1.19 mg min L –1 at pH 7 at 20 °C . Significant differences in free chlorine susceptibility were observed among the tested variants (see the results of Tukey’s HSD test in Table S4).…”

Section: Resultssupporting

confidence: 89%

“…The result of the predicted CT values for 4log inactivation of CVB5 Faulkner (1.02 mg min L −1 ) was consistent with a previous study, reporting 1.15−1.19 mg min L −1 at pH 7 at 20 °C. 50 Significant differences in free chlorine susceptibility were observed among the tested variants (see the results of Tukey's HSD test in Table S4). Free chlorine susceptibility was different by up to 3.4-fold within CVB5.…”

Section: Free Chlorinementioning

confidence: 96%

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Impact of the Heterogeneity in Free Chlorine, UV₂₅₄, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency

Torii

Miura

Itamochi

et al. 2021

Environ. Sci. Technol.

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The disinfection susceptibilities of viruses vary even among variants, yet the inactivation efficiency of a certain virus genotype, species, or genus was determined based on the susceptibility of its laboratory strain. The objectives were to evaluate the variability in susceptibilities to free chlorine, UV254, and ozone among 13 variants of coxsackievirus B5 (CVB5) and develop the model allowing for predicting the overall inactivation of heterogeneous CVB5. Our results showed that the susceptibilities differed by up to 3.4-fold, 1.3-fold, and 1.8-fold in free chlorine, UV254, and ozone, respectively. CVB5 in genogroup B exhibited significantly lower susceptibility to free chlorine and ozone than genogroup A, where the laboratory strain, Faulkner, belongs. The capsid protein in genogroup B contained a lower number of sulfur-containing amino acids, readily reactive to oxidants. We reformulated the Chick–Watson model by incorporating the probability distributions of inactivation rate constants to capture the heterogeneity. This expanded Chick–Watson model indicated that up to 4.2-fold larger free chlorine CT is required to achieve 6-log inactivation of CVB5 than the prediction by the Faulkner strain. Therefore, it is recommended to incorporate the variation in disinfection susceptibilities for predicting the overall inactivation of a certain type of viruses.

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Section: Resultssupporting

confidence: 89%

Section: Free Chlorinementioning

confidence: 96%

Impact of the Heterogeneity in Free Chlorine, UV₂₅₄, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency

Torii

Miura

Itamochi

et al. 2021

Environ. Sci. Technol.

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“…Until the completion of the present study, PMA and PMAxx were not applied to brackish and seawater matrices [ 29 ] and few studies have quantified intact HAdV particles using these dyes in environmental matrices [ 21 , 30 – 32 ]. Only one study used PMAxx dye to analyse HAdV inoculated in phosphate-buffered saline (PBS) solution in thermal and free-chlorine disinfection processes [ 20 ]. The percentage of damaged capsid observed in this study was high, ranging from 69 to 100%, regardless of the concentration of PMAxx and matrix evaluated.…”

Section: Discussionmentioning

confidence: 99%

“…Studies using PMAxx evaluated infectious HAdV particles (serotypes 2, 5, 12, and 40) in samples of phosphate-buffered saline, food (mollusc and sausage), and environmental (soil, sewage, and freshwater from the urban river), thus demonstrating a reduction ranging from 0.42 to 4 log [ 19 , 20 , 22 , 24 , 27 , 45 ]. PMA or PMAxx were also efficient in detecting another viruses, enteric or not, including SARS-CoV-2 with intact capsids in clinical, food, and environmental matrices [ 16 , 31 , 32 , 43 , 46 – 54 ] providing valuable data for environmental surveillance.…”

Section: Discussionmentioning

confidence: 99%

Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR

et al. 2022

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Molecular methodologies providing data on viral concentration and infectivity have been successfully used in environmental virology, supporting quantitative risk assessment studies. The present study aimed to assess human mastadenovirus (HAdV) intact particles using a derivative of propidium monoazide associated with qPCR (PMAxx-qPCR) in aquatic matrices. Initially, different concentrations of PMAxx were evaluated to establish an optimal protocol for treating different naturally contaminated matrices, using 10 min incubation in the dark at 200 rpm at room temperature and 15 min of photoactivation in the PMA-Lite™ LED photolysis device. There was no significant reduction in the quantification of infectious HAdV with increasing concentration of PMAxx used (20 μM, 50 μM, and 100 μM), except for sewage samples. In this matrix, a reduction of 5.01 log of genomic copies (GC)/L was observed from the concentration of 50 μM and revealed 100% HAdV particles with damaged capsids. On the other hand, the mean reduction of 0.51 log in stool samples using the same concentration mentioned above demonstrated 83% of damaged particles eliminated in the stool. Following, 50 μM PMAxx-qPCR protocol revealed a log reduction of 0.91, 0.67, and 1.05 in other samples of raw sewage, brackish, and seawater where HAdV concentration reached 1.47 × 10 4 , 6.81 × 10 2 , and 2.33 × 10 2 GC/L, respectively. Fifty micrometers of PMAxx protocol helped screen intact viruses from different matrices, including sea and brackish water.

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“…Even so, molecular assays do not discriminate between viruses with infective capacity, inactivated viruses, and free genome. To overcome this limitation, viability markers and binding assays have been coupled to qPCR detection to evaluate the integrity of the viral capsid as estimate for viral infectivity also in water samples (Parshionikar et al, 2010;Kim et al, 2011;Coudray-Meunier et al, 2013;Prevost et al, 2016;Randazzo et al, 2016Randazzo et al, , 2018bLópez-Gálvez et al, 2018;Tian et al, 2018;Leifels et al, 2019Leifels et al, , 2020Shirasaki et al, 2020;Canh et al, 2021a). Of note, such approach has been very recently implemented for SARS-CoV-2 detection in wastewater (Canh et al, 2021b;Cuevas-Ferrando et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

Monitoring Human Viral Pathogens Reveals Potential Hazard for Treated Wastewater Discharge or Reuse

et al. 2022

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Wastewater discharge to the environment or its reuse after sanitization poses a concern for public health given the risk of transmission of human viral diseases. However, estimating the viral infectivity along the wastewater cycle presents technical challenges and still remains underexplored. Recently, human-associated crAssphage has been investigated to serve as viral pathogen indicator to monitor fecal impacted water bodies, even though its assessment as biomarker for infectious enteric viruses has not been explored yet. To this end, the occurrence of potentially infectious norovirus genogroup I (GI), norovirus GII, hepatitis A virus (HAV), rotavirus A (RV), and human astrovirus (HAstV) along with crAssphage was investigated in influent and effluent water sampled in four wastewater treatment plants (WWTPs) over 1 year by a PMAxx-based capsid integrity RT-qPCR assay. Moreover, influent and effluent samples of a selected WWTP were additionally assayed by an in situ capture RT-qPCR assay (ISC-RT-qPCR) as estimate for viral infectivity in alternative to PMAxx-RT-qPCR. Overall, our results showed lower viral occurrence and concentration assessed by ISC-RT-qPCR than PMAxx-RT-qPCR. Occurrence of potentially infectious enteric virus was estimated by PMAxx-RT-qPCR as 88–94% in influent and 46–67% in effluent wastewaters with mean titers ranging from 4.77 to 5.89, and from 3.86 to 4.97 log10 GC/L, with the exception of HAV that was sporadically detected. All samples tested positive for crAssphage at concentration ranging from 7.41 to 9.99 log10 GC/L in influent and from 4.56 to 6.96 log10 GC/L in effluent wastewater, showing higher mean concentration than targeted enteric viruses. Data obtained by PMAxx-RT-qPCR showed that crAssphage strongly correlated with norovirus GII (ρ = 0.67, p < 0.05) and weakly with HAstV and RV (ρ = 0.25–0.30, p < 0.05) in influent samples. In effluent wastewater, weak (ρ = 0.27–0.38, p < 0.05) to moderate (ρ = 0.47–0.48, p < 0.05) correlations between crAssphage and targeted viruses were observed. Overall, these results corroborate crAssphage as an indicator for fecal contamination in wastewater but a poor marker for either viral occurrence and viral integrity/infectivity. Despite the viral load reductions detected in effluent compared to influent wastewaters, the estimates of viral infectivity based on viability molecular methods might pose a concern for (re)-using of treated water.

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Suitability of pepper mild mottle virus as a human enteric virus surrogate for assessing the efficacy of thermal or free-chlorine disinfection processes by using infectivity assays and enhanced viability PCR

Cited by 36 publications

References 34 publications

Impact of the Heterogeneity in Free Chlorine, UV₂₅₄, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency

Impact of the Heterogeneity in Free Chlorine, UV₂₅₄, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency

Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR

Monitoring Human Viral Pathogens Reveals Potential Hazard for Treated Wastewater Discharge or Reuse

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Suitability of pepper mild mottle virus as a human enteric virus surrogate for assessing the efficacy of thermal or free-chlorine disinfection processes by using infectivity assays and enhanced viability PCR

Cited by 36 publications

References 34 publications

Impact of the Heterogeneity in Free Chlorine, UV254, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency

Impact of the Heterogeneity in Free Chlorine, UV254, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency

Quantification of infectious Human mastadenovirus in environmental matrices using PMAxx-qPCR

Monitoring Human Viral Pathogens Reveals Potential Hazard for Treated Wastewater Discharge or Reuse

Contact Info

Product

Resources

About

Impact of the Heterogeneity in Free Chlorine, UV₂₅₄, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency

Impact of the Heterogeneity in Free Chlorine, UV₂₅₄, and Ozone Susceptibilities Among Coxsackievirus B5 on the Prediction of the Overall Inactivation Efficiency