Surveillance of SARS-CoV-2 in sewage and wastewater treatment plants in Mexico

Carrillo-Reyes, Julián; Barragán-Trinidad, Martín; Buitrón, Germán

The SARS-CoV-2 virus causing COVID-19 is spread in sewage by the stool of infected individuals, and viral material in sewage can be quantified using molecular tools. This study aimed to monitor the presence of SARS-CoV-2 RNA in sewage in Mexico based on RdRP, S, and N gene analysis. The influent, effluent, and activated sludge from two domestic wastewater treatment plants (WWTP) were evaluated from the early stage of the epidemic to July 2020. Additionally, sampling points in sewer systems were examined, comparing two different RNA-concentration methods: centrifugal ultrafiltration and adsorption-based methods. The adsorption method resulted in RNA titration that was two orders of magnitude higher than with ultrafiltration (up to 3.38 log10 copies RdRP gene/mL of sewage). The surveillance of SARS-CoV-2 RNA in the influent of two WWTP correlated with the cumulative COVID-19 cases in Queretaro city. The higher RNA level in secondary sludge compared to influent suggests that viral RNA becomes concentrated in activated sludge. This result supports SARS-CoV-2 RNA removal in WWTP, where all effluent samples were negative for virus quantification. This work proves that wastewater-based epidemiology is a very valuable tool in developing countries where diagnostic tests for COVID-19 are limited.