Hiv Detection in Wastewater as a New Epidemiological Tool

Background: Wastewater represents a broad, immediate, and unbiased accounting of the pathgens in the population. We aimed to develop methods to track HIV in wastewater utilizing a viral detection pipeline adapted from platforms developed to track SARS-COV-2. Method(s): We used samples from 6 wastewater treatment plants in the Houston area. We focused on regions of higher prevalence and lower prevalence. First, employing wastewater processing and nucleic acid extraction methods described by our group to detect SARS-COV-2, we tested a single high and low prevalence site in triplicate with all 3 primer sets. nucleic acid extracts from HIV and SIV cell culture supernatants were used as controls. Next, in subsequent samples, RT-PCR reactions with detections were subjected to gel electrophoresis to determine the amplified product sizes. To further confirm HIV detection, we sequenced the RT-PCR products and compared the proportion of reads which mapped to the expected amplified product. In a later set of studies, we fractionated samples into supernatant and pellet. We further tested HIV presence by performing whole virome sequencing on the extracts from some samples that produced detections and mapped reads to published genomes. A crAssphage genome was used as a negative control. Result(s): Samples from all sites resulted in signal detection at least once. Only reactions with gag and pol primers appeared to amplify the expected product. Products from the HIV positive control mapped almost exclusively to the HIV genome (97-100% of reads), with a fraction of reads from the SIV negative control doing the same (16-18% of reads). The ltr and pol products did not map the HIV genome while gag products did (34-44% of reads). Among the fractionated sample, in total, 6 supernatant fractions produced no detection compared to 7 of 8 pellet fractions. The whole virome sequencing produced reads that mapped to the HIV genome with at least 8X depth coverage. The sample with the lowest Ct detection (26) yielded HIV coverage several logs greater than those samples with higher Ct detection (37). Reads from all samples mapped to at least 20% of the HIV genome. Conclusion(s): This work provides the first evidence that HIV can be detected in municipal wastewater systems and has the potential to be developed into a new public health tool.

Analysis of the Chemistry of Municipal Wastewaters for Forecasting the Conditions of Urban Population

The quality of wastewater and the socioeconomic aspects of the life of the population are shown to be interrelated. It is proposed to use municipal wastewater to diagnose the health of the population, to assess the feeding preferences of city dwellers, the use of alcohol, tobacco, medicines, and drugs, to assess the exposure of the population to the effect of hazardous chemicals (PAH, pesticides, preserving agents, plasticizers, etc.), and to timely reveal deceases (including COVID-19). The authors proposed several biomarkers, which are recommended to use to monitor the water-resource system as a component of the urban ecosystem.

Beyond COVID: The Value of Wastewater Based Epidemiology (WBE) Monitoring for Municipal and Industrial Systems

The goal of this paper is to demonstrate how Wastewater Based Epidemiology (WBE) can be used after COVID-19 in both Municipal and Industrial wastewater systems to proactively monitor, manage, and avoid risks that could negatively impact the business continuity and resiliency of an organization. The history of WBE will first be reviewed to show how it has been used to maximize public health protection and social well-being while minimizing economic impacts and unintended consequences in public and private settings. The design of a WBE monitoring program for Closed, Semi-Closed, and Open Municipal and Industrial wastewater systems will be evaluated through a couple of case studies. Alignment between WBE programs and an organizations' risk management programs, sustainability goals, and ethical considerations will also be explored. Copyright © 2022 Water Environment Federation.

A Review of the Presence of SARS-CoV-2 in Wastewater: Transmission Risks in Mexico.

The appearance of SARS-CoV-2 represented a new health threat to humanity and affected millions of people; the transmission of this virus occurs through different routes, and one of them recently under debate in the international community is its possible incorporation and spread by sewage. Therefore, the present work's research objectives are to review the presence of SARS-CoV-2 in wastewater throughout the world and to analyze the coverage of wastewater treatment in Mexico to determine if there is a correlation between the positive cases of COVID-19 and the percentages of treated wastewater in Mexico as well as to investigate the evidence of possible transmission by aerosol sand untreated wastewater. Methodologically, a quick search of scientific literature was performed to identify evidence the presence of SARS-CoV-2 RNA (ribonucleic acid) in wastewater in four international databases. The statistical information of the positive cases of COVID-19 was obtained from data from the Health Secretary of the Mexican Government and the Johns Hopkins Coronavirus Resource Center. The information from the wastewater treatment plants in Mexico was obtained from official information of the National Water Commission of Mexico. The results showed sufficient evidence that SARS-CoV-2 remains alive in municipal wastewater in Mexico. Our analysis indicates that there is a low but significant correlation between the percentage of treated water and positive cases of coronavirus r = -0.385, with IC (95%) = (-0.647, -0.042) and p = 0.030; this result should be taken with caution because wastewater is not a transmission mechanism, but this finding is useful to highlight the need to increase the percentage of treated wastewater and to do it efficiently. In conclusions, the virus is present in untreated wastewater, and the early detection of SAR-CoV-2 could serve as a bioindicator method of the presence of the virus. This could be of great help to establish surveillance measures by zones to take preventive actions, which to date have not been considered by the Mexican health authorities. Unfortunately, wastewater treatment systems in Mexico are very fragile, and coverage is limited to urban areas and non-existent in rural areas. Furthermore, although the probability of contagion is relatively low, it can be a risk for wastewater treatment plant workers and people who are close to them.

Wastewater-based epidemiology approach: The learning lessons from COVID-19 pandemic and the development of novel guidelines for future pandemics.

Wastewater-based epidemiology (WBE) provides a comprehensive real-time framework of population attitude and health status. This approach is attracting the interest of medical community and health authorities to monitor the prevalence of a virus (such as the severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) among a community. Indeed, WBE is currently fine-tuning as environmental surveillance tool for coronavirus disease 2019 (COVID-19) pandemic. After a bibliometric analysis conducted to discover the research trends in WBE field, this work aimed to side-by-side compare the conventional method based on clinical testing with WBE approach. Furthermore, novel guidelines were developed to apply the WBE approach to a pandemic. The growing interest on WBE approach for COVID-19 pandemic is demonstrated by looking at the sharp increase in scientific papers published in the last years and at the ongoing studies on viral quantification methods and analytical procedures. The side-by-side comparison highlighted the ability of WBE to identify the hot-spot areas faster than the conventional approach, reducing the costs (e.g., rational use of available resources) and the gatherings at medical centers. Contrary to clinical testing, WBE has the surveillance capacity for preventing the virus resurgence, including asymptomatic contribution, and ensuring the preservation of medical staff health by avoiding the exposure to the virus infection during clinical testing. As extensively reported, the time in collecting epidemiological data is crucial for establishing the prevention and mitigation measures that are essential for curbing a pandemic. The developed guidelines can help to build a WBE system useful to control any future pandemic.

Wastewater-based epidemiology: a new frontier for tracking environmental persistence and community transmission of COVID-19.

Recent research in many parts of the world has pointed towards evidence of SARS-CoV-2 RNA in both treated and raw municipal wastewater discharged by communities. Therefore, concerns regarding it being a possible enteric virus are abundant. Past history of SARS-CoV-1 outbreaks and viral survival information helps in establishing information regarding possible viral infectivity and survival of SARS-CoV-2. The paper examines the existing strategies and techniques including the efficacy of laboratory-based RT-qPCR technique for tracking environmental persistence and community transmission of COVID-19. Analysis of studies targeting untreated and treated wastewater as source of samples is carried out. The analysis shows that untreated samples were mostly positive for SARS-CoV-2 RNA in the target studies. Infectivity estimation from viral load data was found to be about two orders of magnitude higher than actual case data in one of the studies. Additionally, relevant research on environmental survivability of SARS-CoV-2 and possible gaps are examined. Biosensors and excretion metabolite tracking in viral detection are also examined, which hold tremendous importance for future research. Wastewater-based epidemiology (WBE) shows incredible promise in the near future for tracking environmental persistence and community transmission of highly infectious diseases such as SARS-CoV-2. With limited research available on SARS-CoV-2 with regard to WBE, it is imperative that focus be established on the evidence-based targeted studies.

SARS-CoV-2 in municipal wastewater treatment plant, collection network, and hospital wastewater.

The current outbreak of coronavirus disease (COVID-19) has led to creating a public health emergency conditions since 2019. COVID-19, which is caused by SARS-CoV-2, is spread via human-to-human transmission by direct contact or droplets. Through conducting this study, we were looking for detecting SARS-CoV-2 in wastewater produced in Iran country (Ardabil, Nir, Khalkhal, and Kowsar) (wastewater collection network, wastewater treatment plant, and hospital wastewater). In this research, samples (n=76) were collected from influent and effluent of municipal and hospital wastewater treatment plants, and some samples were also collected from Ardabil municipal wastewater manholes. The sampling duration included the white (lower risk of COVID-19) and red (high risk of COVID-19) conditions. Samples were stored at -20 °C for further diagnostic tests. The specific primer and probe real-time reverse transcriptase-polymerase chain reaction (real-time PCR) targeting ORF1ab and N genes (nucleoprotein gene) were applied to detect viral genomes of the SARS-CoV-2 virus in the wastewater samples. Out of 76 samples, a total of 15 samples (19.73%) collected from wastewater in Ardabil province (Ardabil, Nir, Khalkhal, and Kowsar), were positive in terms of SARS-CoV-2. Wastewater epidemiology can facilitate detection of the incidence of pathogens through metropolises, measurement of population prevalence without direct testing, and provision of information to the public health system about the efficiency of intervening efforts. Graphical abstract.

Occurrence and fate of antidepressants in the aquatic environment: Insights into toxicological effects on the aquatic life, analytical methods, and removal techniques

The consumption of antidepressants has increased on a global scale. These medications are frequently prescribed to treat mental health-related disorders and their usage is expected to rise in the future because the COVID-19 pandemic has intensified these problems significantly. These compounds have recently been detected in wastewater treatment plants and surface waters, raising concerns about their potential impacts on the envi-ronment. In this regard, the current review aims to critically evaluate the available information on the worldwide consumption of antidepressants, their occurrence, possible toxicological effects on aquatic organisms, and removal techniques. Several analytical methods for the extraction and quantification of antidepressant com-pounds have also been discussed. Additionally, risk quotients (RQs) have been estimated which indicates that sertraline posed the highest risk (RQ: 4.88) to the aquatic life followed by citalopram (RQ: 1.55) and bupropion (RQ: 1.12). It was observed that the aquatic organisms encountered behavioral, physical, cardiovascular, and reproductive changes after being exposed to antidepressant compounds. Some of these compounds have been satisfactorily removed (>85%) using a sequencing batch reactor with aerobic granulation of sludge. Physico-chemical processes such as photocatalysis, photochemical oxidation, and electrocatalysis exhibited more than 90% degradation efficiency in most cases. Moreover, integrating two or more physicochemical processes improved the treatment efficiency further. This study may help researchers to understand the threats posed by antidepressants to the environment and result in the development of innovative technologies for their removal.

Investigating SARS-CoV-2 RNA in five municipal wastewater treatment plants, hospital wastewater and wastewater collection networks during the COVID-19 pandemic in Ardabil Province, Iran.

Since 2019, the outbreak of coronavirus with acute respiratory symptoms has caused an epidemic worldwide. Transmission of the disease through respiratory droplets was announced as the main mode of transmission in 2020. But in this study, we discussed the method of indirect transmission of the virus through sewage. In this study, effluents related to urban and hospital wastewater treatment plants in 5 regions of Ardabil Province (northwest of Iran) were investigated. In this research, 120 samples were kept in pre-test conditions (temperature -20 degrees Celsius). To identify the viral genome, special primer and chain reaction probe targeting ORF1ab and N (nucleoprotein gene) genes were used. Out of a total of 120 samples, a total of 3 samples were positive. Wastewater epidemiology (WBE) can be considered as a cost-effective method in the diagnosis and prediction of pathogenic agents. And be considered an effective method for decision-making in order to protect the health of citizens.

Sludge-based activated carbon from two municipal sewage sludge precursors for improved secondary wastewater-treatment discharge-effluent

This study investigated the use of sludge-based activated carbon (SBAC) sorbent as an integrated waste-to-resources approach for the removal of contaminants from wastewater. We measured the ability of SBAC sorbents from two types of municipal sewage sludge (SS) precursors (thickened waste SS “TWSS-SBAC” and biosolids “Bio-SBAC”) from a Canadian wastewater treatment plant (WWTP) to stabilise emerging contaminants (ECs) from precursor SS and to remove ECs from the discharged effluent. The ECs were from pharmaceutical and personal care products (PPCPs), including antibiotics, disinfectants, and antibacterial hand-sanitisers and soaps, which were commonly used during the COVID-19 (coronavirus disease of 2019) pandemic. We measured the removal efficacy of Bio-SBAC at two dosages (1g/L and 10g/L) and TWSS-SBAC at one dosage (1g/L) via 30-min batch adsorption tests for eleven PPCPs at mean concentrations of 2–2337ng/L in the discharged effluent, and compared the results with those of other techniques and sorbents reported in literature. At both dosages, Bio-SBAC removed PPCPs, including four blood regulator compounds that have been extensively used since the pandemic outbreak (furosemide, gemfibrozil, glyburide, and warfarin), with their levels decreasing below the detection limit. The percentage removal for ibuprofen, 2-hydroxy-ibuprofen, and naproxen were 91.6–99.8% using 1g/L. The antimicrobial compounds triclosan and triclocarban were completely removed at both dosages. Ninety-nine percentage of bisphenol A was removed at 1g/L dosage and was completely removed at 10g/L. TWSS-SBAC showed similar performance as Bio-SBAC in removing PPCPs from the final effluent to improve the quality of wastewater discharged from a WWTP.

A pilot study of wastewater monitoring for SARS-CoV-2 in New Zealand

Surveillance of municipal wastewater for RNA of SARS-CoV-2, the causative agent of coronavirus disease (COVID-19), is well documented around the world. However, unlike most countries where wastewater surveillance was initially employed during 2020, New Zealand was in the fortunate position of having very few COVID-19 cases, generally confined to Managed Isolation and Quarantine facilities. As such, the prevalence of SARS-CoV-2 RNA in wastewater was likely much lower than seen in other countries. A nine-week pilot study was undertaken to assess the feasibility of detecting SARSCoV-2 RNA in wastewater in New Zealand. Wastewater from 18 catchments across New Zealand was monitored, including six that contained Managed Isolation and Quarantine facilities. Testing both in regions known to have COVID-19 cases and regions where detection was not expected (catchments not containing Managed Isolation and Quarantine facilities) allowed the sensitivity and specificity of detection methods to be assessed. SARS-CoV-2 RNA was detected in seven out of the nine weeks of this study in the Auckland South Western Interceptor catchment, which contained a dedicated isolation facility to which confirmed cases from Auckland, Hamilton and Rotorua were transferred. In weeks two and three of sampling, SARS-CoV-2 RNA was detected in the Christchurch catchment. This coincided with up to 14 COVID-19 cases likely to be shedding high levels of virus (PCR Cq value < 20) in the Managed Isolation and Quarantine facilities. Samples from the seven other weeks were negative despite up to 35 infected cases present at any one time. However, on any of these test dates eight cases or fewer had a PCR Cq value < 30 and were within 10 days of symptom onset or positive PCR test date. Sample inhibition and non-specificity were not observed to be issues. The results of this pilot study underpinned recommendations that wastewater monitoring for SARS-CoV-2 RNA be incorporated as a surveillance tool in New Zealand's COVID-19 response.

Exploring opportunities for sewage testing on cargo ships as a tool to screen seafarers for COVID-19

Stringent border controls and surveillance measures have been an essential part of the New Zealand Government's response to the coronavirus disease 2019 (COVID-19) pandemic, particularly its elimination strategy, which requires all infected individuals arriving at the border to be identified and isolated to prevent incursions and community transmission. The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) virus that causes COVID-19 is shed in the faeces and other bodily secretions of infected people, and municipal wastewater-based surveillance for SARS-CoV-2 has been extensively utilised at both localised and community scales. This review article explores whether testing the sewage from cargo ships arriving at the New Zealand maritime border could be used in a similar way, serving as a screening mechanism to help protect the border. Whilst considerable knowledge gaps remain regarding the prevalence, onset and kinetics of faecal shedding of SARS-CoV-2, the available data suggest approximately half of infected individuals shed detectable levels of RNA in their stool, with the onset of shedding starting anywhere between the first and fifth week of illness. Engineering and health and safety considerations are also discussed. We conclude that testing sewage from ships is unlikely to detect cases of COVID-19 amongst crew with the reliability required by an elimination strategy, particularly given individual-level nasopharyngeal testing is readily administered to the relatively small crew aboard these vessels. However, other opportunities for testing sewage from ships for SARS-CoV-2 may exist, such as when border settings are relaxed and/or individual-level testing is not practical or warranted (e.g., cruise ships), or in surveillance for other pathogens.

Application of human RNase P normalization for the realistic estimation of SARS-CoV-2 viral load in wastewater: A perspective from Qatar wastewater surveillance.

The apparent uncertainty associated with shedding patterns, environmental impacts, and sample processing strategies have greatly influenced the variability of SARS-CoV-2 concentrations in wastewater. This study evaluates the use of a new normalization approach using human RNase P for the logic estimation of SARS-CoV-2 viral load in wastewater. SARS-CoV-2 variants outbreak was monitored during the circulating wave between February and August 2021. Sewage samples were collected from five major wastewater treatment plants and subsequently analyzed to determine the viral loads in the wastewater. SARS-CoV-2 was detected in all the samples where the wastewater Ct values exhibited a similar trend as the reported number of new daily positive cases in the country. The infected population number was estimated using a mathematical model that compensated for RNA decay due to wastewater temperature and sewer residence time, and which indicated that the number of positive cases circulating in the population declined from 765,729 ± 142,080 to 2,303 ± 464 during the sampling period. Genomic analyses of SARS-CoV-2 of thirty wastewater samples collected between March 2021 and April 2021 revealed that alpha (B.1.1.7) and beta (B.1.351) were among the dominant variants of concern (VOC) in Qatar. The findings of this study imply that the normalization of data allows a more realistic assessment of incidence trends within the population.

Design and Validation of Sample Splitting Protocol for Comparison of SARS-CoV-2 Quantification in Wastewater

Evaluations of analytical performance through interlaboratory comparisons and proficiency tests are underway globally for biomolecular-based methods [e.g., reverse-transcription quantitative polymerase chain reaction (RT-qPCR)] used in the surveillance of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in wastewater. These evaluations often rely on sharing a common reference wastewater sample that is split among participating laboratories. A known quantity of recovery surrogates can be introduced to the wastewater matrix by the coordinating laboratory as an exogenous control in a spike-and-recovery approach;however, split-sample comparisons are increasingly performed to evaluate in situ quantities of SARS-CoV-2 genetic signal native to the sample due to the lack of a universally accepted recovery surrogate of SARS-CoV-2. A reproducible procedure that minimizes the variability of SARS-CoV-2 genetic signal among split wastewater aliquots is therefore necessary to facilitate the method comparisons, especially when a large number of aliquots are required. Emerging literature has suggested that SARS-CoV-2 genetic signal in wastewater is linked to the solids fraction. Accordingly, a protocol that allows for equal distribution of solids content evenly among wastewater aliquots was also likely to facilitate even distribution of the SARS-CoV-2 genetic signal. Based on this premise, we reviewed existing sample splitting apparatus and approaches used for solids-based parameters in environmental samples. A portable batch reactor was designed, comprised of readily accessible materials and equipment. This design was validated through splitting of real wastewater samples collected from a municipal wastewater treatment facility serving a population with reported cases of COVID-19. This work applies well-established solid-liquid mixing theory and concepts that are likely unfamiliar to molecular microbiologists and laboratory analysts, providing (1) a prototype adaptable for a range of sample quantities, aliquot sizes, microbial targets, and water matrices;and (2) a pragmatic demonstration of critical considerations for design and validation of a reproducible and effective sample splitting protocol.

The Influence of Municipal Wastewater Treatment Technologies on the Biological Stabilization of Sewage Sludge: A Systematic Review

Various wastewater treatment technologies are available today and biological processes are predominantly used in these technologies. Increasing wastewater treatment systems produces large amounts of sewage sludge with variable quantities and qualities, which must be properly managed. Anaerobic and aerobic digestion and composting are major strategies to treat this sludge. The main indicators of biological stabilization are volatile fatty acids (VFAs), volatile solids (VS), the carbon/nitrogen (C/N) ratio, humic substances (HS), the total organic carbon (TOC), the carbon dioxide (CO2) evolution rate, the specific oxygen uptake rate (SOUR), and the Dewar test;however, different criteria exist for the same indicators. Although there is no consensus for defining the stability of sewage sludge (biosolids) in the research and regulations reviewed, controlling the biological degradation, vector attraction, and odor determines the biological stabilization of sewage sludge. Because pollutants and pathogens are not completely removed in biological stabilization processes, further treatments to improve the quality of biosolids and to ensure their safe use should be explored.

Identification of Key Factors for Urban—Industrial Water Reuse: A Multi-Criteria Analysis Case Study

The industrial reuse of existing municipal wastewater treatment plant (WWTP) effluent can play a major role in improving water security in urbanized regions facing scarcity. As the complexity of engineered direct water reuse is related to various economic, technical, legal, social, environmental, and public health aspects, multi-criteria analysis (MCA) is a feasible decision-making tool in this context. The present work aimed to establish the relevant key factors for the application of MCA, wherever plant planning, design, and construction did not previously consider reuse practices. The adopted methodology considers the proposition and valuation of key criteria, based on the existing literature, expert consultations, statistical analysis, and the application of MCA to a real municipal WWTP located in Campinas city (São Paulo State, Brazil). The 13 proposed criteria encompass multiple categories, and their relevance is demonstrated, given the high significance frequencies assigned. The best values are related to effluent quality, health risks, and treatment reliability, in addition to environmental costs and benefits. The application of those criteria in Cooperative Game Theory (CGT) and Compromise Programming (CP) methods is proved to be suitable, considering the characteristics of the studied area (i.e., highly urbanized with a history of water scarcity). Among nine surveyed end-users, the first position in the hierarchy corresponds to the largest industries with the shortest distance from the WWTP.

A graphene-printed paper electrode for determination of H2O2in municipal wastewater during the COVID-19 pandemic

Recently, hydrogen peroxide (H2O2) has been used as a disinfectant in sanitizers for cleaning hands, and solid surfaces of hospitals, offices and homes to prevent the spread of the COVID-19 virus. The effluents from domestic, hospital and municipal waste should be monitored for their H2O2 content to avoid the entry of this toxic pollutant into the ecosystem. Therefore, we developed a low-cost graphene (Gr)-printed paper electrode for determination of H2O2 using cyclic voltammetry (CV). An office inkjet-printer and Gr nano-ink stabilized with ethyl cellulose (EC) were used for the fabrication of printed paper electrodes (PPEs) to determine H2O2 quantitatively. A stable Gr-EC nano-ink (2%) with viscosity and surface tension values of 12 mPa S-1 and 35 mN M-1, respectively, was formulated to obtain conductive electrodes. A wide linear range (2 μM-25 mM) with a better limit of detection (0.28 μM) for the determination of H2O2 was obtained when the Gr-EC/PPE was used as a working electrode. Further, the Gr-EC/PPE was successfully employed for analysis of H2O2 in wastewater. The electrochemical determination of H2O2 using the Gr-EC/PPE as an electrode in CV is rapid, economical, flexible and eco-friendly when compared with previously reported methods.

Development of a rapid pre-concentration protocol and a magnetic beads-based RNA extraction method for SARS-CoV-2 detection in raw municipal wastewater

In this work, a rapid and simplified method for extracting SARS-CoV-2 RNA from whole wastewater using a magnetic beads-based protocol is presented. The described method involves the centrifugation of a 50-mL aliquot of raw wastewater influent for 5 min to obtain a 500-μL pellet, which is eluted with 2 mL of a Tween®20-based elution buffer;1 mL of the elute is extracted for RNA using a direct magnetic bead-based extraction method. RNA recovery was examined in several bench-scale experiments using heat-inactivated SARS-CoV-2 (HI-SCV-2) spiked into raw wastewater to assess the effects of different solids pellet : buffer ratios, inhibition mitigation strategies, and varying levels of total suspended solids. When the method was assessed using an influent wastewater sample known to contain SARS-CoV-2, the viral signal was detected in all five biological replicates, whereas direct extraction of 1-mL aliquots of the raw wastewater resulted in a 40% viral detection rate. The experimental method limit of detection (MLOD) using HI-SCV-2 spiked into raw wastewater was 50 GU mL−1 with a 95% limit of detection. Using the described protocol, the presence of SARS-CoV-2 RNA was verified in wastewater collected from wastewater treatment facilities (WWTFs) in Atlantic Canada over a period of 15 weeks during the rise and fall of a COVID-19 outbreak. This method is effective and rapid and could provide potential application for laboratories with limited resources. Of approximately 50 methods that have been developed for measuring SARS-CoV-2 in wastewater referenced in the literature, this is the first to advance a robust magnetic beads-based RNA extraction technique from whole wastewater without extensive sample pre-treatment. The novel application of this method in the rapid extraction of SARS-CoV-2 RNA from municipal wastewater is an indispensable tool to potentially understand COVID-19 infection occurrence within communities.

Systematic assessment of SARS-CoV-2 virus in wastewater, rivers and drinking water - A catchment-wide appraisal.

Human coronaviruses (HCoVs) attracted attention in 2002 with the severe acute respiratory syndrome (SARS) outbreak, caused by the SARS-CoV virus (mortality rate 9.6%), and gained further notoriety in 2012 with the Middle East respiratory syndrome (MERS) (mortality rate 34.3%). Currently, the world is experiencing an unprecedented crisis due to the COVID-19 global pandemic, caused by the SARS-CoV-2 virus in 2019. The virus can pass to the faeces of some patients, as was the case of SARS-CoV and MERS-CoV viruses. This suggests that apart from the airborne (droplets and aerosols) and person-to-person (including fomites) transmission, the faecal-oral route of transmission could also be possible for HCoVs. In this eventuality, natural water bodies could act as a virus reservoir of infection. Here, the temporospatial migration and attenuation of the SARS-CoV-2 virus in municipal wastewater, the receiving environment, and drinking water is evaluated, using the polymerase chain reaction (PCR), in the South African setting. SARS-CoV-2 viral RNA was identified in raw wastewater influent but was below the detection limit in the latter treatment stages. This suggests that the virus decays from as early as primary treatment and this could be attributed to wastewater's hydraulic retention time (2-4 h), composition, and more importantly temperature (>25 °C). Therefore, the probability of SARS-CoV-2 virus transportation in water catchments, in the eventuality that the virus remains infective in wastewater, appears to be low in the South African setting. Finally, catchment-wide monitoring offers a snapshot of the status of the catchment in relation to contagious viruses and can play a pivotal role in informing the custodians and downstream water users of potential risks embedded in water bodies.

Antiviral drug Umifenovir (Arbidol) in municipal wastewater during the COVID-19 pandemic: Estimated levels and transformation.

An indole derivative umifenovir (Arbidol) is one of the most widely used antiviral drugs for the prevention and treatment of COVID-19 and some other viral infections. The purpose of the present study was to shed light on the transformation processes of umifenovir in municipal wastewater, including disinfection with active chlorine, as well as to assess the levels of the antiviral drug and its metabolites entering and accumulating in natural reservoirs under conditions of the SARS-CoV-2 pandemic. The combination of high-performance liquid chromatography with electrospray ionization high-resolution mass-spectrometry and inductively coupled plasma mass spectrometry was used for tentative identification and quantification of umifenovir and its transformation products in model reaction mixtures and real samples of wastewater, river water, biological sludge and bottom sediments taken at the wastewater treatment plant in Arkhangelsk, a large cultural and industrial center at the Russian North. Laboratory experiments allowed identifying fifteen bromine-containing transformation products, forming at the initial stages of the chlorination and fourteen classic volatile and semi volatile disinfection by-products with bromoform as the dominant one. Chlorinated derivatives are only the minor disinfection by-products forming by substitution of alkylamine group in the aromatic ring. The schemes of umifenovir transformation in reactions with dissolved oxygen and sodium hypochlorite are proposed. Two established primary transformation products formed by oxidation of the thioether group to sulfoxide and elimination of thiophenol were detected in noticeable concentrations in the wastewater together with their precursor. The level of umifenovir reached 1.3 mg kg-1 in the sludge and municipal wastewater treat contained 1 µg L-1 of that drug, while its removal during biological wastewater treatment was about 40%. Pronounced accumulation of umifenovir and its transformation products in biological sludge and bottom sediments of natural reservoirs may be a source of the future secondary pollution of the environment.