Abstract

Enterovirus 70 (EV70) is an emerging viral pathogen that remains viable in final treated effluent. Solar irradiation is, therefore, explored as a low-cost natural disinfection strategy to mitigate potential concerns. EV70 was exposed to simulated sunlight for 24 h at a fluence rate of 28.67 J/cm2/h in three different water matrices, namely, phosphate-buffered saline (PBS), treated wastewater effluent, and chlorinated effluent. In the presence of sunlight, EV70 decreased in infectivity by 1.7 log, 1.0 log, and 1.3 log in PBS, effluent, and chlorinated effluent, respectively. Irradiated EV70 was further introduced to host cell lines and was unable to infect the cell lines. In contrast, EV70 in dark microcosms replicated to titers 13.5, 3.3, and 4.2 times the initial inoculum. The reduction in EV70 infectivity was accompanied by a reduction in viral binding capacity to Vero cells. In addition, genome sequencing analysis revealed five nonsynonymous nucleotide substitutions in irradiated viruses after 10 days of infection in Vero cells, resulting in amino acid substitutions: Lys14Glu in the VP4 protein, Ala201Val in VP2, Gly71Ser in VP3, Glu50Gln in VP1, and Ile47Leu in 3Cpro. Overall, solar irradiation resulted in EV70 inactivation and an inhibition of viral activity in all parameters studied.

Highlights

  • Climate change, urbanization, and increasing global population have placed considerable pressure on freshwater supplies [1,2,3]

  • This indicates that the existing disinfection procedures employed are not adequate to provide safe water for reuse, and there exists a need to explore the efficacy of solar irradiation as a possible additional disinfection strategy against enterovirus 70 (EV70)

  • The pellet was resuspended in 50 mL of sterile 1× phosphate-buffered saline (PBS) or 0.45 μm-filtered effluent or chlorinated effluent wastewater collected from the Wastewater Treatment Plant of King Abdullah

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Summary

Introduction

Urbanization, and increasing global population have placed considerable pressure on freshwater supplies [1,2,3]. A species within the Picornaviridae family that has not been studied in this aspect is enterovirus 70 (EV70) These viruses are mainly transmitted by the fecal-oral route and cause gastroenteritis. Infectious EV70 has been detected in the effluents of several WWTPs globally [6,23,24,25] This indicates that the existing disinfection procedures employed are not adequate to provide safe water for reuse, and there exists a need to explore the efficacy of solar irradiation as a possible additional disinfection strategy against enterovirus 70 (EV70). All of the irradiated viruses in this study failed to replicate in cell culture, providing a strong endorsement of sunlight as a low-cost natural disinfection strategy

Cells and Viruses
Simulated Solar Inactivation Trials
Virus Inactivation Kinetics Evaluated by Means of Infectious Assay
Virus Inactivation Kinetics Evaluated by Means of RNA Concentration Decay
Growth Curve Analysis
Virus Absorption Assay
EV70 Genome Sequencing
IIII RT enzyme
Viral Inactivation Upon Solar Irradiation
RNA Decay
Irradiated EV70 Displays Inhibited Viral Replication
Irradiated
Discussion
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