Tracking the occurrence and fate of human bocavirus 2 in municipal wastewater from a small city

Abstract

Background Human Bocavirus 2 (HBoV2) is an emerging enteric virus frequently detected in wastewater, yet its environmental fate, persistence, and treatment remain poorly understood. Improved characterization of HBoV2 is important for advancing wastewater-based epidemiology and understanding viral transmission dynamics. Methods and Results A year-long wastewater surveillance study was conducted from December 2023 to December 2024 in a small urban wastewater treatment system. Viral concentrations were quantified using digital PCR (dPCR), decay kinetics were assessed using controlled mesocosm experiments, and phylogenetic relationships were evaluated through NP1 gene sequencing. HBoV2 was detected in 93.8% of influent samples (average 4.76 log₁₀ genome copies per liter (GC/L) ) with seasonal peaks in summer and fall. Phylogenetic analysis revealed clustering with globally circulating HBoV2 lineages. Decay experiments indicated slow viral degradation (k = 0.10 d⁻¹), comparable to other non-enveloped DNA viruses including Adeno-Associated Virus type 2 (AAV-2), Adenovirus 41 (AdV41), and Human Polyomavirus (HPyV). Additionally, HBoV2 persisted through secondary treatment, with no significant reduction observed between influent and secondary effluent samples. Conclusion Our findings indicate HBoV2 as highly prevalent and environmentally stable in a small urban city’s wastewater and that it is resistant to conventional treatment. These findings highlight HBoV2’s potential role in gastrointestinal infections worldwide and the need for further monitoring of this environmental contaminant.

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