Quantification of SARS-CoV-2 in wastewater samples from hospitals treating COVID-19 patients during the first wave of the pandemic in Brazil.

The COVID-19 pandemic has caused a global health crisis, and wastewater-based epidemiology (WBE) has emerged as an important tool to assist public health decision-making. Recent studies have shown that the SARS-CoV-2 RNA concentration in wastewater samples is a reliable indicator of the severity of the pandemic for large populations. However, few studies have established a strong correlation between the number of infected people and the viral concentration in wastewater due to variations in viral shedding over time, viral decay, infiltration, and inflow. Herein we present the relationship between the number of COVID-19-positive patients and the viral concentration in wastewater samples from three different hospitals (A, B, and C) in the city of Belo Horizonte, Minas Gerais, Brazil. A positive and strong correlation between wastewater SARS-CoV-2 concentration and the number of confirmed cases was observed for Hospital B for both regions of the N gene (R = 0.89 and 0.77 for N1 and N2, respectively), while samples from Hospitals A and C showed low and moderate correlations, respectively. Even though the effects of viral decay and infiltration were minimized in our study, the variability of viral shedding throughout the infection period and feces dilution due to water usage for different activities in the hospitals could have affected the viral concentrations. These effects were prominent in Hospital A, which had the smallest sewershed population size, and where no correlation between the number of defecations from COVID-19 patients and viral concentration in wastewater was observed. Although we could not determine trends in the number of infected patients through SARS-CoV-2 concentrations in hospitals' wastewater samples, our results suggest that wastewater monitoring can be efficient for the detection of infected individuals at a local level, complementing clinical data.

Long-term monitoring of SARS-CoV-2 RNA in sewage samples from specific public places and STPs to track COVID-19 spread and identify potential hotspots.

Coronavirus pandemic started in March 2020 and since then has caused millions of deaths worldwide. Wastewater-based epidemiology (WBE) can be used as an epidemiological surveillance tool to track SARS-CoV-2 dissemination and provide warning of COVID-19 outbreaks. Considering that there are public places that could be potential hotspots of infected people that may reflect the local epidemiological situation, the presence of SARS-CoV-2 RNA was analyzed by RT-qPCR for approximately 16 months in sewage samples from five public places located in the metropolitan area of Belo Horizonte, MG, Brazil: the sewage treatment plant of Confins International Airport (AIR), the main interstate bus terminal (BUS), an upscale shopping centre (SHC1), a popular shopping centre (SHC2) and a university institute (UNI). The results were compared to those of the influent sewage of the two main sewage treatment plants of Belo Horizonte (STP1 and STP2). Viral monitoring in the STPs proved to be an useful regional surveillance tool, reflecting the trends of COVID-19 cases. However, the viral concentrations in the samples from the selected public places were generally much lower than those of the municipal STPs, which may be due to the behaviour of the non-infected or asymptomatic people, who are likely to visit these places relatively more than the symptomatic infected ones. Among these places, the AIR samples presented the highest viral concentrations and concentration peaks were observed previously to local outbreaks. Therefore, airport sewage monitoring can provide an indication of the regional epidemiological situation. For the other places, particularly the UNI, the results suggested a greater potential to detect the infection and trace cases especially among employees and regular attendees. Taken together, the results indicate that for a regular and permanent sentinel sewage surveillance the sewage from STPs, AIR and UNI could be monitored.

Virtual screening of molecular databases for potential inhibitors of the NSP16/NSP10 methyltransferase from SARS-CoV-2.

COVID-19 is a disease caused by the SARS-CoV-2 virus and represents one of the greatest health problems that humanity faces at the moment. Therefore, efforts have been made with the objective of seeking therapies that could be effective in combating this problematic. In the search for ligands, computational chemistry plays an essential role, since it allows the screening of thousands of molecules on a given target, in order to save time and money for the in vitro or in vivo pharmacological stage. In this paper, we perform a virtual screening by docking looking for potential inhibitors of the NSP16-NSP10 protein dimer (methyltransferase) from SARS-CoV-2, by evaluating a homemade databank of molecules found in plants of the Caatinga Brazilian biome, compounds from ZINC online molecular database, as well as structural analogues of the enzymatic cofactor s-adenosylmethionine (SAM) and a known inhibitor in the literature, sinefungin (SFG), provided at PubChem database. All the evaluated sets presented molecules that deserve attention, highlighting four compounds from ZINC as the most promising ligands. These results contribute to the discovery of new molecular hits, in the search of potential agents against SARS-CoV-2 virus, still unveiling a pathway that can be used in combined therapies.

Reduction and liquid-solid partitioning of SARS-CoV-2 and adenovirus throughout the different stages of a pilot-scale wastewater treatment plant.

Investigating waterborne viruses is of great importance to minimizing risks to public health. Viruses tend to adsorb to sludge particles from wastewater processes by electrostatic and hydrophobic interactions between virus, aquatic matrix, and particle surface. Sludge is often re-used in agriculture; therefore, its evaluation is also of great interest to public health. In the present study, a pilot scale system treating real domestic wastewater from a large city in Brazil was used to evaluate the removal, the overall reduction, and liquid-solid partitioning of human adenovirus (HAdV), the novel coronavirus (SARS-CoV-2) and fecal indicators (F-specific coliphages and E. coli). The system consists of a high-rate algal pond (HRAP) post-treating the effluent of an upflow anaerobic sludge blanket (UASB) reactor. Samples were collected from the influent and effluent of each unit, as well as from the sludge of the UASB and from the microalgae biomass in the HRAP. Pathogens and indicators were quantified by quantitative polymerase chain reaction (qPCR) (for HAdV), qPCR with reverse transcription (RTqPCR) (for SARS-CoV-2), the double agar plaque assay (for coliphages), and the most probable number (MPN) method (for E. coli). The removal and overall reduction of HAdV and SARS-CoV-2 was greater than 1-log10. Almost 60% of remaining SARS-CoV-2 RNA and more than 70% of remaining HAdV DNA left the system in the sludge, demonstrating that both viruses may have affinity for solids. Coliphages showed a much lower affinity to solids, with only 3.7% leaving the system in the sludge. The system performed well in terms of the removal of organic matter and ammoniacal nitrogen, however tertiary treatment would be necessary to provide further pathogen reduction, if the effluent is to be reused in agriculture. To our knowledge, this is the first study that evaluated the reduction and partitioning of SARS-CoV-2 and HAdV through the complete cycle of a wastewater treatment system consisting of a UASB reactor followed by HRAPs.

Assessing spatial distribution of COVID-19 prevalence in Brazil using decentralised sewage monitoring.

Brazil has become one of the epicentres of the COVID-19 pandemic, with cases heavily concentrated in large cities. Testing data is extremely limited and unreliable, which restricts health authorities' ability to deal with the pandemic. Given the stark demographic, social and economic heterogeneities within Brazilian cities, it is important to identify hotspots so that the limited resources available can have the greatest impact. This study shows that decentralised monitoring of SARS-CoV-2 RNA in sewage can be used to assess the distribution of COVID-19 prevalence in the city. The methodology developed in this study allowed the identification of hotspots by comprehensively monitoring sewers distributed through Belo Horizonte, Brazil's third largest city. Our results show that the most vulnerable neighbourhoods in the city were the hardest hit by the pandemic, indicating that, for many Brazilians, the situation is much worse than reported by official figures.