Revealing the Epidemiology of Human Bocavirus and <i>Streptococcus pneumoniae</i> through Wastewater Surveillance

Human bocavirus (HBoV) and Streptococcus pneumoniae pose significant public health concerns, particularly for children under age five. While wastewater-based epidemiology has proven successful in monitoring the prevalence of infectious diseases in communities, its potential for surveillance of HBoV and S. pneumoniae has not been thoroughly explored. We first optimized the Efficient and Practical virus Identification System with Enhanced Sensitivity for Membrane (EPISENS-M) method, which was originally designed for RNA virus detection, to detection viral and bacterial DNA efficiently. Our findings revealed that the EPISENS-M method, without DNase enzyme treatment and using a filtration volume of 75 mL, yields a high detection of tested DNA (e.g., crAssphage and Bacteroides HF183) in wastewater samples. We then applied the protocol for surveillance of HBoV and S. pneumoniae in wastewater samples collected weekly from two wastewater treatment plants in Sapporo, Japan, for a one-year period. HBoV was detected in the majority of wastewater samples (94%), with a noticeable peak during the summer. S. pneumoniae was highly prevalent (frequency: 100%), but without seasonal variations in concentration levels. This corresponds with clinical surveillance data from recent years in Japan, underscoring the importance of wastewater surveillance for understanding the epidemiology of HBoV and S. pneumoniae in communities.