Abstract

F-specific RNA bacteriophages (FRNAPHs) can be used to indicate water contamination and the fate of viruses in wastewater treatment plants (WWTPs). However, the occurrence of FRNAPH strains in WWTPs is relatively unknown, whereas FRNAPH genotypes (GI–GIV) are well documented. This study investigated the diversity of infectious FRNAPH strains in wastewater treatment and disinfection processes using cell culture combined with next-generation sequencing (integrated culture–NGS (IC–NGS)). A total of 32 infectious strains belonging to FRNAPH GI (nine strains), GI-JS (two strains), GII (nine strains), GIII (seven strains), and GIV (five strains) were detected in wastewater samples. The strains of FRNAPH GI and GII exhibited greater resistance to wastewater treatment than those of GIII. The IC–NGS results in the disinfected samples successfully reflected the infectivity of FRNAPHs by evaluating the relationship between IC–NGS results and the integrated culture–reverse-transcription polymerase chain reaction combined with the most probable number assay, which can detect infectious FRNAPH genotypes. The diversity of infectious FRNAPH strains in the disinfected samples indicates that certain strains are more resistant to chlorine (DL52, GI-JS; T72, GII) and ultraviolet (T72, GII) disinfection. It is possible that investigating these disinfectant-resistant strains could reveal effective mechanisms of viral disinfection.

Highlights

  • F-specific RNA bacteriophages (FRNAPHs), which are known to infect Escherichia coli that expressF pili, have a single-stranded RNA genome enclosed in an icosahedral capsid measuring 20–30 nm in diameter

  • The aim of this study was to investigate the diversity of infectious FRNAPH strains in wastewater treatment and disinfection processes using IC–Next-generation sequencing (NGS)

  • A total of 32 FRNAPH strains were successfully detected in wastewater samples by integrated culture–NGS (IC–NGS) (Figure 1)

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Summary

Introduction

F-specific RNA bacteriophages (FRNAPHs), which are known to infect Escherichia coli that expressF pili, have a single-stranded RNA genome enclosed in an icosahedral capsid measuring 20–30 nm in diameter. FRNAPH behavior, abundance, and survival in the environment including during water treatment are similar to those of human enteric viruses [1,2,4,5,6]. They serve as potential indicators of water contamination and the fates of viruses in aquatic environments and wastewater treatment plants (WWTPs) [4,5,6]. Each FRNAPH genotype has a different fate in WWTPs [6,7,8,9,10] and a different resistance to disinfection [11,12,13]. GI is the dominant genotype in the secondary effluent of WWTPs because of its higher resistance to wastewater treatment relative to other

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