SARS-CoV-2 RNA Detection in Wastewater and Its Effective Correlation with Clinical Data during the Outbreak of COVID-19 in Salamanca.

Wastewater treatment plants (WWTPs) are the final stage of the anthropogenic water cycle where a wide range of chemical and biological markers of human activity can be found. In COVID-19 disease contexts, wastewater surveillance has been used to infer community trends based on viral abundance and SARS-CoV-2 RNA variant composition, which has served to anticipate and establish appropriate protocols to prevent potential viral outbreaks. Numerous studies worldwide have provided reliable and robust tools to detect and quantify SARS-CoV-2 RNA in wastewater, although due to the high dilution and degradation rate of the viral RNA in such samples, the detection limit of the pathogen has been a bottleneck for the proposed protocols so far. The current work provides a comprehensive and systematic study of the different parameters that may affect the detection of SARS-CoV-2 RNA in wastewater and hinder its quantification. The results obtained using synthetic viral RNA as a template allow us to consider that 10 genome copies per µL is the minimum RNA concentration that provides reliable and consistent values for the quantification of SARS-CoV-2 RNA. RT-qPCR analysis of wastewater samples collected at the WWTP in Salamanca (western Spain) and at six pumping stations in the city showed that below this threshold, positive results must be confirmed by sequencing to identify the specific viral sequence. This allowed us to find correlations between the SARS-CoV-2 RNA levels found in wastewater and the COVID-19 clinical data reported by health authorities. The close match between environmental and clinical data from the Salamanca case study has been confirmed by similar experimental approaches in four other cities in the same region. The present methodological approach reinforces the usefulness of wastewater-based epidemiology (WBE) studies in the face of future pandemic outbreaks.

Model-based evaluation of a trickling filter facility upgrade to biological nutrient removal.

This article presents the feasibility evaluation and preliminary design of a wastewater treatment plant upgrade supported by simulation. The existing facility was based on trickling filters, and the objective of the upgrade was to achieve nutrients removal. The proposed solution modifies the existing primary clarifier to host an anaerobic-anoxic suspended growth reactor, which is an alternative that, to our knowledge, has not been proposed or explored so far. The trickling filters would remain as aerobic reactors. In this study, the novel treatment scheme has been assessed for the first time, through model simulations. The modified treatment train was simulated, showing that the anoxic zone is able to denitrify satisfactorily achieving the required effluent nitrogen concentration. However, to promote biological phosphorus removal, an additional aerobic zone combined with a bypass of activated sludge from the anoxic zone to the first trickling filter is needed, in order to provide aerobic conditions to the phosphate accumulating organisms. Several combinations of additional aerobic volume and sludge bypass flowrate were found to successfully achieve both nitrogen and phosphorus removal, using the existing facilities without the need for new reactors neither implementing modifications that could put the trickling filters' physical integrity at risk. The novel treatment scheme could be applied in other cases with similar flowsheet in the same context.