Capturing the SARS-CoV-2 infection pyramid within the municipality of Rotterdam using longitudinal sewage surveillance (preprint)

Background: Despite high vaccination rates in the Netherlands, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to circulate. Longitudinal sewage monitoring was implemented along with the notification of cases as two parts of the surveillance pyramid to validate the use of sewage surveillance for monitoring SARS-CoV-2, as an early warning tool, and to measure the effect of interventions. Methods: Sewage samples were collected from nine neighborhoods from September 2020 to November 2021, and compared with reported cases. Comparative analysis and modeling were performed to understand the correlation between wastewater and case trends. Findings: Using high resolution sampling, normalization of wastewater SARS-CoV-2 concentrations and normalization of reported positive tests for testing delay and intensity, the incidence of reported positive tests could be modeled based on sewage data, and trends in both surveillance systems coincided. The high collinearity implied that high levels of viral shedding around the onset of disease largely determines SARS-CoV-2 levels in wastewater and the observed relation was independent of SARS-CoV-2 variants and vaccination levels. Interpretation: Wastewater surveillance can accurately display SARS-CoV-2 dynamics for small and large locations, and is sensitive enough to measure small variations in the number of infected individuals within or between neighborhoods. With the transition to a post-acute phase of the pandemic, continued sewage surveillance can help to keep sight on reemergence, but continued pyramid validation studies are needed to assess the predictive value of sewage surveillance with new variants. Funding: Horizon H2020, Adessium Foundation, STOWA, TKI, Ministry of Health, Welfare and Sport

Monitoring SARS-CoV-2 circulation and diversity through community wastewater sequencing (preprint)

The current SARS-CoV-2 pandemic has rapidly become a major global health problem for which public health surveillance is crucial to monitor virus spread. Given the presence of viral RNA in feces in around 40% of infected persons, wastewater-based epidemiology has been proposed as an addition to disease-based surveillance to assess the spread of the virus at the community level. Here we have explored the possibility of using next-generation sequencing (NGS) of sewage samples to evaluate the diversity of SARS-CoV-2 at the community level from routine wastewater testing, and compared these results with the virus diversity in patients from the Netherlands and Belgium. Phylogenetic analysis revealed the presence of viruses belonging to the most prevalent clades (19A, 20A and 20B) in both countries. Clades 19B and 20C were not identified, while they were present in clinical samples during the same period. Low frequency variant (LFV) analysis showed that some known LFVs can be associated with particular clusters within a clade, different to those of their consensus sequences, suggesting the presence of at least 2 clades within a single sewage sample. Additionally, combining genome consensus and LFV analyses we found a total of 57 unique mutations in the SARS-CoV-2 genome which have not been described before. In conclusion, this work illustrates how NGS analysis of wastewater can be used to approximate the diversity of SARS-CoV-2 viruses circulating in a community.