Longitudinal monitoring of SARS-CoV-2 in wastewater using viral genetic markers and the estimation of unconfirmed COVID-19 cases.

In this study, wastewater-based surveillance was carried out to establish the correlation between SARS-CoV-2 viral RNA concentrations in wastewater and the incidence of corona virus disease 2019 (COVID-19) from clinical testing. The influent wastewater of three major water reclamation facilities (WRFs) in Northern Nevada, serving a population of 390,750, was monitored for SARS-CoV-2 viral RNA gene markers, N1 and N2, from June 2020 through September 2021. A total of 614 samples were collected and analyzed. The SARS-CoV-2 concentrations in wastewater were observed to peak twice during the study period. A moderate correlation trend between coronavirus disease 2019 (COVID-19) incidence data from clinical testing and SARS-CoV-2 viral RNA concentrations in wastewater was observed (Spearman r = 0.533). This correlation improved when using weekly average SARS-CoV-2 marker concentrations of wastewater and clinical case data (Spearman r = 0.790), presumably by mitigating the inherent variability of the environmental dataset and the effects of clinical testing artifacts (e.g., reporting lags). The research also demonstrated the value of wastewater-based surveillance as an early warning signal for early detection of trends in COVID-19 incidence. This was accomplished by identifying that the reported clinical cases had a stronger correlation to SARS-CoV-2 wastewater monitoring data when they were estimated to lag 7-days behind the wastewater data. The results aided local decision makers in developing strategies to manage COVID-19 in the region and provide a framework for how wastewater-based surveillance can be applied across localities to enhance the public health monitoring of the ongoing pandemic.

Spatial and temporal variability and data bias in wastewater surveillance of SARS-CoV-2 in a sewer system.

The response to disease outbreaks, such as SARS-CoV-2, can be constrained by a limited ability to measure disease prevalence early at a localized level. Wastewater based epidemiology is a powerful tool identifying disease spread from pooled community sewer networks or at influent to wastewater treatment plants. However, this approach is often not applied at a granular level that permits detection of local hot spots. This study examines the spatial patterns of SARS-CoV-2 in sewage through a spatial sampling strategy across neighborhood-scale sewershed catchments. Sampling was conducted across the Reno-Sparks metropolitan area from November to mid-December of 2020. This research utilized local spatial autocorrelation tests to identify the evolution of statistically significant neighborhood hot spots in sewershed sub-catchments that were identified to lead waves of infection, with adjacent neighborhoods observed to lag with increasing viral RNA concentrations over subsequent dates. The correlations between the sub-catchments over the sampling period were also characterized using principal component analysis. Results identified distinct time series patterns, with sewersheds in the urban center, outlying suburban areas, and outlying urbanized districts generally following unique trends over the sampling period. Several demographic parameters were identified as having important gradients across these areas, namely population density, poverty levels, household income, and age. These results provide a more strategic approach to identify disease outbreaks at the neighborhood level and characterized how sampling site selection could be designed based on the spatial and demographic characteristics of neighborhoods.