High-throughput sequencing reveals the virome characteristics of wastewater in Gansu Province, China from 2024 to 2026

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Emerging and re-emerging infectious diseases continue to pose a persistent challenge to global public health security. As an unbiased and noninvasive population-level surveillance approach, wastewater-based epidemiology has emerged as an urban-scale monitoring tool for proactively capturing the community disease burden. In this study, metagenomic and targeted next-generation sequencing were employed to preliminarily characterize wastewater viral communities in Gansu Province from 2024 to 2026. Across the three annual sampling groups, the Simpson diversity index calculated from metagenomic taxonomic profiles showed a significant negative correlation with sampling year. Despite viral enrichment, more than 97% of sequencing reads were assigned to bacteria, likely reflecting residual nonviral nucleic acids and limitations in metagenomic classification. The wastewater virome exhibited a complex host-association spectrum, within which bacteriophages and plant-associated viruses displayed relatively stable community structures and high relative abundances. Among human pathogens, astroviruses were consistently detected at high relative abundances across all samples. Human astrovirus 1 accounted for the largest proportion, and a nearly complete genome sequence of the extraintestinal infection-associated astrovirus MLB2 was recovered. Targeted sequencing further revealed a highly structured environmental enterovirus profile dominated by the species Enterovirus betacoxsackie , followed by Enterovirus alphacoxsackie , whereas Enterovirus coxsackiepol was detected at an extremely low abundance and Enterovirus deconjuncti was not detected. In addition to several common viruses associated with hand, foot, and mouth disease, rare viruses, including coxsackievirus A19 and enterovirus B83, were successfully captured. Echovirus 11 was identified as the predominant enterovirus serotype in current wastewater surveillance in Gansu Province. In summary, this study provides a preliminary characterization of the viral community profiles in wastewater samples from Gansu Province, demonstrates the effectiveness of wastewater-based epidemiology for population-level surveillance, and underscores the need to establish a coordinated defense network integrating clinical case surveillance with environmental monitoring.

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