Evaluation of SARS-CoV-2 RNA Presence in Treated and Untreated Hospital Sewage.

Wastewater-based epidemiology (WBE) is a potential approach for determining the viral prevalence in a community. In the wake of the COVID-19 pandemic, researchers have begun to pay close attention to the presence of SARS-COV-2 RNA in various wastewaters. The potential for detecting SARS-CoV-2 RNA in hospital sewage could make it an invaluable resource for epidemiological studies. In this regard, two specialized hospitals dedicated to COVID-19 patients were chosen for this investigation. Both hospitals utilize the same wastewater treatment systems. The influent and effluents of the two hospitals were sampled in May and June of 2021, and the samples were evaluated for their chemical properties. According to the findings of this study, the wastewater qualities of the two studied hospitals were within the standard ranges. The sewage samples were concentrated using ultrafiltration and PEG precipitation techniques. The E and S genes were studied with RT-qPCR commercial kits. We found E gene of SARS-CoV-2 in 83.3% (5/6) and 66.6% (4/6) of wastewater samples from hospital 1 and hospital 2, respectively, using ultrafiltration concentration method. Wastewater samples taken after chlorine treatment accounted for 16.6% of all positive results. In addition, due to the small sample size, there was no significant correlation (p > 0.05) between the presence of SARS-CoV-2 in wastewater and the number of COVID-19 cases. Hospitals may be a source of SARS-CoV-2 pollution, thus it is important to monitor and enhance wastewater treatment systems to prevent the spread of the virus and safeguard the surrounding environment.

Analytical methods for the determination of antidepressants, antipsychotics, benzodiazepines and their metabolites through wastewater-based epidemiology

The wastewater-based epidemiology approach, popular for estimating illicit drug use, has been used to evaluate lifestyle habits such as alcohol, tobacco and caffeine consumption, health biomarkers including pharmaceuticals, plasticizers and flame retardants, and recently to track SARS-CoV-2 at the population level. Equally, the number of WBE studies investigating psychoactive pharmaceuticals such as antidepressants, antipsychotics and benzodiazepines has also increased, which can be connected to the overall growth of psychological disorders worldwide. This review aims to discuss novelties in sampling techniques and analytical methodologies, including sample preparation and analysis, developed for estimating the consumption of psychoactive pharmaceuticals in defined populations. Seventy-four peer-reviewed studies monitoring psychoactive pharmaceutical consumption published since 2010 have been systematically reviewed. Its findings show that a broad range of bioanalytical methodologies is used to simultaneously measure several antidepressants, antipsychotics, benzodiazepines, and their metabolites from influent wastewater samples in low concentrations and different time periods. The application of WBE commenting on the temporal and spatial variations worldwide, showing widespread consumption, is also discussed. Despite much progress and excellent studies, there remains a need for research, and deeper knowledge is needed to reduce method uncertainty, especially since excretion rates, their transformation, and in-sewer and in-sample stability for many psychoactive pharmaceutical biomarkers are not available.

Rapid transition between SARS-CoV-2 variants of concern Delta and Omicron detected by monitoring municipal wastewater from three Canadian cities.

Monitoring the communal incidence of COVID-19 is important for both government and residents of an area to make informed decisions. However, continuous reliance on one means of monitoring might not be accurate because of biases introduced by government policies or behaviours of residents. Wastewater surveillance was employed to monitor concentrations of SARS-CoV-2 RNA in raw influent wastewater from wastewater treatment plants serving three Canadian Prairie cities with different population sizes. Data obtained from wastewater are not directly influenced by government regulations or behaviours of individuals. The means of three weekly samples collected using 24 h composite auto-samplers were determined. Viral loads were determined by RT-qPCR, and whole-genome sequencing was used to charaterize variants of concern (VOC). The dominant VOCs in the three cities were the same but with different proportions of sub-lineages. Sub-lineages of Delta were AY.12, AY.25, AY.27 and AY.93 in 2021, while the major sub-lineage of Omicron was BA.1 in January 2022, and BA.2 subsequently became a trace-level sub-variant then the predominant VOC. When each VOC was first detected varied among cities; However, Saskatoon, with the largest population, was always the first to present new VOCs. Viral loads varied among cities, but there was no direct correlation with population size, possibly because of differences in flow regimes. Population is one of the factors that affects trends in onset and development of local outbreaks during the pandemic. This might be due to demography or the fact that larger populations had greater potential for inter- and intra-country migration. Hence, wastewater surveillance data from larger cities can typically be used to indicate what to expect in smaller communities.

Site Specific Relationships between COVID-19 Cases and SARS-CoV-2 Viral Load in Wastewater Treatment Plant Influent.

Wastewater based epidemiology (WBE) has become an important tool during the COVID-19 pandemic, however the relationship between SARS-CoV-2 RNA in wastewater treatment plant influent (WWTP) and cases in the community is not well-defined. We report here the development of a national WBE program across 28 WWTPs serving 50% of the population of Scotland, including large conurbations, as well as low-density rural and remote island communities. For each WWTP catchment area, we quantified spatial and temporal relationships between SARS-CoV-2 RNA in wastewater and COVID-19 cases. Daily WWTP SARS-CoV-2 influent viral RNA load, calculated using daily influent flow rates, had the strongest correlation (ρ > 0.9) with COVID-19 cases within a catchment. As the incidence of COVID-19 cases within a community increased, a linear relationship emerged between cases and influent viral RNA load. There were significant differences between WWTPs in their capacity to predict case numbers based on influent viral RNA load, with the limit of detection ranging from 25 cases for larger plants to a single case in smaller plants. SARS-CoV-2 viral RNA load can be used to predict the number of cases detected in the WWTP catchment area, with a clear statistically significant relationship observed above site-specific case thresholds.

Prevalence of SARS-CoV-2 genes in water reclamation facilities: From influent to anaerobic digester.

Several treatment plants were sampled for influent, primary clarifier sludge, return activated sludge (RAS), and anaerobically digested sludge throughout nine weeks during the summer of the COVID-19 pandemic. Primary clarifier sludge had a significantly higher number of SARS-CoV-2 gene copy number per liter (GC/L) than other sludge samples, within a range from 1.0 × 105 to 1.0 × 106 GC/L. Gene copy numbers in raw influent significantly correlated with gene copy numbers in RAS in Silver Creek (p-value = 0.007, R2 = 0.681) and East Canyon (p-value = 0.009, R2 = 0.775) WRFs; both of which lack primary clarifiers or industrial pretreatment processes. This data indicates that SARS-CoV-2 gene copies tend to partition into primary clarifier sludges, at which point a significant portion of them are removed through sedimentation. Furthermore, it was found that East Canyon WRF gene copy numbers in influent were a significant predictor of daily cases (p-value = 0.0322, R2 = 0.561), and gene copy numbers in RAS were a significant predictor of weekly cases (p-value = 0.0597, R2 = 0.449). However, gene copy numbers found in primary sludge samples from other plants significantly predicted the number of COVID-19 cases for the following week (t = 2.279) and the week after that (t = 2.122) respectively. These data indicate that SARS-CoV-2 extracted from WRF biosolids may better suit epidemiological monitoring that exhibits a time lag. It also supports the observation that primary sludge removes a significant portion of SARS-CoV-2 marker genes. In its absence, RAS can also be used to predict the number of COVID-19 cases due to direct flow through from influent. This research represents the first of its kind to thoroughly examine SARS-CoV-2 gene copy numbers in biosolids throughout the wastewater treatment process and the relationship between primary, return activated, and anaerobically digested sludge and reported positive COVID-19 cases.

The first detection of SARS-CoV-2 RNA in the wastewater of Tehran, Iran.

Following the official announcement of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide pandemic spread by WHO on March 11, 2020, more than 300,000 COVID-19 cases reported in Iran resulting in approximately 17,000 deaths as of August 2, 2020. In the present survey, we investigated the presence of SARS-CoV-2 RNA in raw and treated wastewater samples in Tehran, Iran. Untreated and treated wastewater samples were gathered from four wastewater treatment plants over a month period from June to July 2020. Firstly, an adsorption-elution concentration method was tested using an avian coronavirus (infectious bronchitis virus, IBV). Then, the method was effectively employed to survey the presence of SARS-CoV-2 genome in influent and effluent wastewater samples. SARS-CoV-2 RNA was found in 8 out of 10 treated wastewater samples utilizing a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) test to detect ORF1ab and N genes. Moreover, the rate of positivity in wastewater samples increased in last sample collection that shows circulation of SARS-CoV-2 was increased among the population. In addition, the high values detected in effluent wastewater from local wastewater treatment plants have several implications in health and ecology that should be further assessed.

Analytical methodologies for the detection of SARS-CoV-2 in wastewater: Protocols and future perspectives.

In March 2020 the World Health Organization announced a pandemic outbreak. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative pathogen for the coronavirus disease-19 (COVID-19) pandemic. The authorities worldwide use clinical science to identify infected people, but this approach is not able to track all symptomatic and asymptomatic cases due to limited sampling capacity of the testing laboratories. This drawback is eliminated by the Wastewater-Based Epidemiology (WBE) approach. In this review, we summarized the peer-reviewed published literature (available as of September 28, 2020), in the field of WBE. The commonly used steps (sampling, storage, concentration, isolation, detection) of the analytical protocols were identified. The potential limitations of each stage of the protocols and good practices were discussed. Finally, new methods for the efficient detection of SARS-CoV-2 were proposed.