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Tobacco and alcohol consumption remain priority public health issues worldwide. As participation in population-based surveys has fallen, it is increasingly challenging to estimate accurately the prevalence of alcohol and tobacco use. Wastewater-based epidemiology (WBE) is an alternative approach for estimating substance use at the population level that does not rely on survey participation. This study examined spatio-temporal patterns in nicotine (a proxy for tobacco) and alcohol consumption in the Australian population via WBE. Daily wastewater samples (n=164) were collected at 18 selected wastewater treatment plants across Australia, covering approximately 45% of the total population. Nicotine and alcohol metabolites in the samples were measured using liquid chromatography-tandem mass spectrometry. Daily consumption of nicotine and alcohol and its associated uncertainty were computed using Monte Carlo simulations. Nationwide daily average and weekly consumption of these two substances were extrapolated using ordinary least squares and mixed effect models. Nicotine and alcohol consumption was observed in all communities. Consumption of these substances in rural towns was three-to-four times higher than in urban communities. The spatial consumption pattern of these substances was consistent across the monitoring periods in 2014-2015. Nicotine metabolites significantly reduced by 14-25% (p=0.001-0.008) (2014-2015) in some catchments. Alcohol consumption remained constant over the studied periods. Strong weekly consumption patterns were observed for alcohol but not nicotine. Nationwide, the daily average consumption per person (aged 15-79 years) was estimated at about 2.5 cigarettes and 1.3-2.0 standard drinks (weekday-weekend) of alcohol. These estimates were close to the sale figure and apparent consumption respectively. Wastewater-based epidemiology is a feasible method for objectively evaluating the geographic, temporal and weekly profiles of nicotine and alcohol consumption in different communities nationally.

Wastewater-based epidemiology is a new approach to monitor drug abuse. It involves collecting wastewater, analysis of residues of drugs or its metabolites in wastewater, and back-calculation of drug consumption by taking into account wastewater flow, stability of drug target residues in wastewater, and excretion rates of drugs/metabolites. Wastewater-based epidemiology has the advantages of being inexpensive and yielding more consistent and near real-time results. It has the great potential to supplement the existing drug monitoring methods. It can be used to build large-scale (regional, national, or even continental) monitoring networks that would yield spatial patterns and temporal trends in drug abuse. This paper described in detail the principle and procedures of this wastewater-based approach. Application of this approach across the globe was also reviewed. The uncertainties involved in the approach and knowledge gaps were identified. Finally, necessity, benefits, and feasibility to set up nation or province-wide monitoring networks based on wastewater analysis in China were discussed.

Wastewater-based epidemiology/wastewater surveillance has been a topic of significant interest over the last year due to its application in SARS-CoV-2 surveillance to track prevalence of COVID-19 in communities. Although SARS-CoV-2 surveillance has been applied in more than 50 countries to date, the application of this surveillance has been largely focused on relatively affluent urban and peri-urban communities. As such, there is a knowledge gap regarding the implementation of reliable wastewater surveillance in small and rural communities for the purpose of tracking rates of incidence of COVID-19 and other pathogens or biomarkers. This study examines the relationships existing between SARS-CoV-2 viral signal from wastewater samples harvested from an upstream pumping station and from an access port at a downstream wastewater treatment lagoon with the community's COVID-19 rate of incidence (measured as percent test positivity) in a small, rural community in Canada. Real-time quantitative polymerase chain reaction (RT-qPCR) targeting the N1 and N2 genes of SARS-CoV-2 demonstrate that all 24-h composite samples harvested from the pumping station over a period of 5.5 weeks had strong viral signal, while all samples 24-h composite samples harvested from the lagoon over the same period were below the limit of quantification. RNA concentrations and integrity of samples harvested from the lagoon were both lower and more variable than from samples from the upstream pumping station collected on the same date, indicating a higher overall stability of SARS-CoV-2 RNA upstream of the lagoon. Additionally, measurements of PMMoV signal in wastewater allowed normalizing SARS-CoV-2 viral signal for fecal matter content, permitting the detection of actual changes in community prevalence with a high level of granularity. As a result, in sewered small and rural communities or low-income regions operating wastewater lagoons, samples for wastewater surveillance should be harvested from pumping stations or the sewershed as opposed to lagoons., Graphical abstract Unlabelled Image

Illicit drug use is complex, hidden and often highly stigmatized behaviour, which brings a vast challenge for drug surveillance systems. Drug consumption can be estimated by measuring human excretion products in untreated wastewater, known as wastewater-based epidemiology (WBE). Over the last decade, the application of wastewater-based epidemiology to monitor illicit drug loads increased and WBE is currently applied on a global scale. Studies from over the globe are evaluated with regard to their sampling method, analytical accuracy and consumption calculation, aiming to further reduce relevant uncertainties in order to make reliable comparisons on a global level. Only a limited number is identified as high-quality studies, so further standardization of the WBE approach for illicit drugs is desired especially with regard to the sampling methodology. Only a fraction of the reviewed papers explicitly reports uncertainty ranges for their consumption data. Studies which had the highest reliability are recently published, indicating an improvement in reporting WBE data. Until now, WBE has not been used in large parts of Africa, nor in the Middle East and Russia. An overview of consumption data across the continents on commonly studied drugs (cocaine, MDMA, amphetamine and methamphetamine) is provided. Overall, high consumption rates are confirmed in the US, especially for cocaine and methamphetamine, while relatively low illicit drug consumption is reported in Asia.

Performance- and image-enhancing drug (PIED) misuse is a significant public health issue. Currently, seizure data, surveys, anti-doping testing, and needle service provider data are used to estimate PIED use in populations. These methods are time consuming, single point-in-time measurements that often consist of small sample sizes and do not truly capture PIED prevalence. Wastewater-based epidemiology (WBE) has been used globally to assess and monitor licit and illicit drug consumption within the general community. This method can objectively cover large populations as well as specific subpopulations (gyms, music festivals, prisons), and has potential as a complementary monitoring method for PIED use. Information obtained through WBE could be used to aid public health authorities in developing targeted prevention and education programmes. Research on PIED analysis in wastewater is limited and presents a significant gap in the literature. The focus is on anabolic steroids, and one steroid alternative currently growing in popularity; selective androgenic receptor modulators. This encompasses medical uses, addiction, prevalence, user typology, and associated public health implications. An overview of WBE is described including its benefits, limitations and potential as a monitoring method for PIED use. A summary of previous work in this field is presented. Finally, we summarise gaps in the literature, future perspectives, and recommendations for monitoring PIEDs in wastewater.

Wastewater-based epidemiology (WBE) has been regarded as a potential tool for the prevalence estimation of coronavirus disease 2019 (COVID-19) in the community. However, the application of the conventional back-estimation approach is currently limited due to the methodological challenges and various uncertainties. This study systematically performed meta-analysis for WBE datasets and investigated the use of data-driven models for the COVID-19 community prevalence in lieu of the conventional WBE back-estimation approach. Three different data-driven models, i.e. multiple linear regression (MLR), artificial neural network (ANN), and adaptive neuro fuzzy inference system (ANFIS) were applied to the multi-national WBE dataset. To evaluate the robustness of these models, predictions for sixteen scenarios with partial inputs were compared against the actual prevalence reports from clinical testing. The performance of models was further validated using unseen data (data sets not included for establishing the model) from different stages of the COVID-19 outbreak. Generally, ANN and ANFIS models showed better accuracy and robustness over MLR models. Air and wastewater temperature played a critical role in the prevalence estimation by data-driven models, especially MLR models. With unseen datasets, ANN model reasonably estimated the prevalence of COVID-19 (cumulative cases) at the initial phase and forecasted the upcoming new cases in 2–4 days at the post-peak phase of the COVID-19 outbreak. This study provided essential information about the feasibility and accuracy of data-driven estimation of COVID-19 prevalence through the WBE approach., Graphical abstract Unlabelled Image

The benefits of wastewater-based epidemiology (WBE) for tracking the viral load of SARS-CoV-2, the causative agent of COVID-19, have become apparent since the start of the pandemic. However, most sampling occurs at the wastewater treatment plant influent and therefore monitors the entire catchment, encompassing multiple municipalities, and is conducted using quantitative polymerase chain reaction (qPCR), which only quantifies one target. Sequencing methods provide additional strain information and also can identify other pathogens, broadening the applicability of WBE to beyond the COVID-19 pandemic. Here we demonstrate feasibility of sampling at the neighborhood or building complex level using qPCR, targeted sequencing, and untargeted metatranscriptomics (total RNA sequencing) to provide a refined understanding of the local dynamics of SARS-CoV-2 strains and identify other pathogens circulating in the community. We demonstrate feasibility of tracking SARS-CoV-2 at the neighborhood, hospital, and nursing home level with the ability to detect one COVID-19 positive out of 60 nursing home residents. The viral load obtained was correlative with the number of COVID-19 patients being treated in the hospital. Targeted wastewater-based sequencing over time demonstrated that nonsynonymous mutations fluctuate in the viral population. Clades and shifts in mutation profiles within the community were monitored and could be used to determine if vaccine or diagnostics need to be adapted to ensure continued efficacy. Furthermore, untargeted RNA sequencing identified several other pathogens in the samples. Therefore, untargeted RNA sequencing could be used to identify new outbreaks or emerging pathogens beyond the COVID-19 pandemic., Graphical abstract Unlabelled Image

The mass load of pharmaceuticals in the municipal wastewater based on wastewater-based epidemiology (WBE) is a good indication of population consumption in the catchment. After successful application of illicit drugs’ estimation, this method holds the potential to measure the geographical and temporal consumption of prescription medicines. In this study, we investigated the occurrence of four non-steroidal anti-inflammatory drugs (NSAIDs), acetaminophen (ACM), diclofenac (DCF), ibuprofen (IBU) and naproxen (NPX), in two wastewater treatment plants in Guangzhou City, China and compared the spatial and temporal consumption variation of them. Over a period of 28 days’ sampling, the detection frequency of ACM, DCF, IBU, and NPX in the influent of two wastewater treatment plants (WWTPs) in Guangzhou City were 91%, 66%, 100%, and 95%, and their concentrations were up to 128, 131, 372, and 324 ng/L, respectively. No significant inter-catchment difference was observed regarding the per capita mass load in the two WWTPs investigated. A literature review which covered 160 WWTPs in 18 countries was conducted to compare the population normalized mass load of four commonly used NSAIDs. ACM had the highest population normalized mass loads (29–17,430 mg/d/1000 inhabitants) and DCF had the lowest population normalized mass load (6.5–628 mg/d/1000 inhabitants) in the catchments located in 18 countries. The mass loads of selected NSAIDs in China were lower than those in European and North American. ACM and IBU consumptions were at least 2 times higher in winter than that in summer, in contrast, DCF and NPX consumptions had no significant seasonal variation.

Wastewater based epidemiology is a potential early warning tool for the detection of COVID-19 outbreak. Sewage surveillance for SARS-CoV-2 RNA was introduced in Hungary after the successful containment of the first wave of the pandemic to forecast the resurge of infections. Three wastewater treatment plants servicing the entire population (1.8 million) of the capital, Budapest were sampled weekly. 24 h composite (n = 44) and grab samples (n = 21) were concentrated by an in-house flat sheet membrane ultrafiltration method. The efficiency and reproducibility of the method was comparable to those previously published. SARS-CoV-2 RNA was quantified using RT-qPCR of the N gene. The first positive signal in sewage was detected 2 weeks before the rise in case numbers. Viral concentration and volume-adjusted viral load correlated to the weekly new cases from the same week and the rolling 7-day average of active cases in the subsequent week. The correlation was more pronounced in the ascending phase of the outbreak, data was divergent once case numbers plateaued. Wastewater surveillance was found to be effective in predicting the second wave of the outbreak in Hungary. Data indicated that even relatively low frequency (weekly) sampling is useful and at the same time, cost effective tool in outbreak detection., Graphical abstract Unlabelled Image

Monitoring of COVID-19 infections within communities via wastewater-based epidemiology could provide a cost-effective alternative to clinical testing. This approach, however, still requires improvement for its efficient application. In this paper, we present the use of wastewater-based epidemiology in monitoring COVID-19 infection dynamics in the KwaZulu-Natal province of South Africa, focusing on four wastewater treatment plants for 14 weeks. The SARS-CoV-2 viral load in influent wastewater was determined using droplet digital PCR, and the number of people infected was estimated using published models as well as using a modified model to improve efficiency. On average, viral loads ranged between 0 and 2.73 × 105 copies/100 ml, 0–1.52 × 105 copies/100 ml, 3 × 104–7.32 × 105 copies/100 ml and 1.55 × 104–4.12 × 105 copies/100 ml in the four wastewater treatment plants studied. The peak in viral load corresponded to the reported COVID-19 infections within the districts where these catchments are located. In addition, we also observed that easing of lockdown restrictions by authorities corresponded with an increase in viral load in the untreated wastewater. Estimation of infection numbers based on the viral load showed that a higher number of people could potentially be infected, compared to the number of cases reported based on clinical testing. The findings reported in this paper contributes to the field of wastewater-based epidemiology for COVID-19 surveillance, whilst highlighting some of the challenges associated with this approach, especially in developing countries., Graphical abstract Unlabelled Image

The aim of this study was to set up a simple protocol to concentrate SARS-CoV-2 from sewage, which can be implemented in laboratories with minimal equipment resources. The method avoids the need for extensive purification steps and reduces the concentration of potential inhibitors of RT-qPCR contained in sewage. The concentration method consists of a single step, in which a small volume (40 mL) of sewage sample is incubated with polyaluminum chloride (PAC)(0.00045 N Al3+ final concentration). Virus particles adsorbed to the precipitate are collected by low-speed centrifugation, after which the recovered pellet is resuspended with a saline buffer. PAC-concentrated samples are stable for at least one week at 4 °C. Therefore, they may be sent refrigerated to a diagnosis center for RNA extraction and RT-qPCR for SARS-CoV-2 RNA detection if the lab does not have such capabilities. The PAC concentration method produced an average shift of 4.5-units in quantification cycle (Cq) values compared to non-concentrated samples, indicating a 25-fold increase in detection sensitivity. The lower detection limit corresponded approximately to 100 viral copies per ml. Kappa index indicated substantial agreement between PAC and polyethylene glycol (PEG) precipitation protocols (k = 0.688, CI 0.457-0.919). This low-cost concentration protocol could be useful to aid in the monitoring of community circulation of SARS-CoV-2, especially in low- and middle-income countries, which do not have massive access to support from specialized labs for sewage surveillance.

Wastewater based epidemiology (WBE) has drawn significant attention as an early warning tool to detect and predict the trajectory of COVID-19 cases in a community, in conjunction with public health data. This means of monitoring for outbreaks has been used at municipal wastewater treatment centers to analyze COVID-19 trends in entire communities, as well as by universities and other community living environments to monitor COVID-19 spread in buildings. Sample concentration is crucial, especially when viral abundance in raw wastewater is below the threshold of detection by RT-qPCR analysis. We evaluated the performance of a rapid ultrafiltration-based virus concentration method using InnovaPrep Concentrating Pipette (CP) Select and compared this to the established electronegative membrane filtration (EMF) method. We evaluated sensitivity of SARS-CoV-2 quantification, surrogate virus recovery rate, and sample processing time. Results suggest that the CP Select concentrator is more efficient at concentrating SARS-CoV-2 from wastewater compared to the EMF method. About 25% of samples that tested negative when concentrated with the EMF method produced a positive signal with the CP Select protocol. Increased recovery of the surrogate virus control using the CP Select confirms this observation. We optimized the CP Select protocol by adding AVL lysis buffer and sonication, to increase the recovery of virus. Sonication increased Bovine Coronavirus (BCoV) recovery by 19%, which seems to compensate for viral loss during centrifugation. Filtration time decreases by approximately 30% when using the CP Select protocol, making this an optimal choice for building surveillance applications where quick turnaround time is necessary., Graphical abstract Unlabelled Image

Wastewater-based epidemiology (WBE) has emerged as a useful tool in the fight to track and contain COVID-19 spread within communities. One of the motives behind COVID-19 WBE efforts is the potential for 'early warning' of either the onset of disease in a new setting or changes in trends in communities where disease is endemic. Many initial reports of the early warning potential of WBE have relied upon retrospective sample analysis, and delays in WBE analysis and reporting should be considered when evaluating the early warning potential of WBE that enable public health action. Our purpose in this manuscript is to establish a framework to critique the potential of WBE to serve as an early warning system, with special attention to the onset of viral shedding and the differential between results reporting for WBE and clinical testing. While many uncertainties remain regarding both COVID-19 clinical presentation and technical factors influencing WBE results, our analysis suggests at most a modest lead time interval ranging from six days for clinical testing to four days for WBE during community-level wastewater surveillance where clinical testing is accessible on-demand with a rapid time to results. This potential lead time for WBE subsequently increases in settings with limited clinical testing capacity or utilization. Care should be taken when reporting 'early detection' of COVID-19 disease trends via WBE to consider underlying causes (e.g., clinical testing lag or delayed result reporting) to avoid misrepresenting WBE potential.

Wastewater-based epidemiology has potential as an early-warning tool for determining the presence of COVID-19 in a community. The University of Arizona (UArizona) utilized WBE paired with clinical testing as a surveillance tool to monitor the UArizona community for SARS-CoV-2 in near real-time, as students re-entered campus in the fall. Positive detection of virus RNA in wastewater lead to selected clinical testing, identification, and isolation of three infected individuals (one symptomatic and two asymptomatic) that averted potential disease transmission. This case study demonstrated the value of WBE as a tool to efficiently utilize resources for COVID-19 prevention and response. Thus, WBE coupled with targeted clinical testing was further conducted on 13 dorms during the course of the Fall semester (Table 3). In total, 91 wastewater samples resulted in positive detection of SARS-CoV-2 RNA that successfully provided an early-warning for at least a single new reported case of infection (positive clinical test) among the residents living in the dorm. Overall, WBE proved to be an accurate diagnostic for new cases of COVID-19 with an 82.0% positive predictive value and an 88.9% negative predictive value. Increases in positive wastewater samples and clinical tests were noted following holiday-related activities. However, shelter-in-place policies proved to be effective in reducing the number of daily reported positive wastewater and clinical tests. This case study provides evidence for WBE paired with clinical testing and public health interventions to effectively contain potential outbreaks of COVID-19 in defined communities., Graphical abstract Unlabelled Image

Alcohol use has increased in the last years and is a priority public health issue worldwide. Wastewater-based epidemiology (WBE) is a reliable approach for estimating substance use at the population level and can complement population surveys and sales statistics to obtain objective and updated information on alcohol use in a community. This work reports the first nationwide study of alcohol consumption conducted in Italy by WBE, covering 11.4% of the Italian population. A total of 264 raw wastewater samples (24-h composite) were collected from 17 Italian cities in 2013–2014 and one large city (Milan) over a 7-year period (2013–2019). Besides of normal days, special periods/events (i.e. Halloween, Milan Fashion Week and summer holidays) were studied in Milan. The urinary biomarker of ethanol (namely, ethyl sulphate) was measured by liquid chromatography-tandem mass spectrometry and was used to estimate alcohol consumption. Results were statistically evaluated to assess spatial and temporal differences and were compared with data from other epidemiological indicators. In 2013, alcohol consumption was significantly higher in small cities than in large ones but no significant differences were observed in relation to the geographical position. An increase in alcohol use was observed in seven large cities from 2013 to 2014. The 7-year monitoring in Milan showed no marked changes, except for remarkable increases in 2014 and 2019. Alcohol consumption significantly increased (ca. 25%) during weekends in Italy and in Milan, and during special periods/events, for instance summer holidays and Halloween. WBE estimates were in agreement with national prevalence data, demonstrating that WBE is suitable for objectively evaluating geographic, annual and weekly patterns of alcohol consumption at local and national scales. WBE information would be helpful to implement and evaluate measures for reducing alcohol consumption and preventing the related health risks.

Surveillance of SARS-CoV-2 and organic tracers (OTs) were conducted in the community wastewater of Chennai city and the suburbs, South India, during partial and post lockdown phases (August–September 2020) as a response to the coronavirus disease 2019 (COVID-19) pandemic. Wastewater samples were collected from four sewage treatment plants (STPs), five sewage pumping stations (SPSs) and at different time intervals from a suburban hospital wastewater (HWW). Four different methods of wastewater concentrations viz., composite (COM), supernatant (SUP), sediment (SED), and syringe filtration (SYR) were subjected to quantitative real time-polymerase chain reaction (qRT-PCR). Unlike HWW, STP inlet, sludge and SPS samples were found with higher loading of SARS-CoV-2 by SED followed by SUP method. Given the higher levels of dissolved and suspended solids in STPs and SPSs over HWW, we suspect that this enveloped virus might exhibit the tendency of higher partitioning in solid phase. Cycle threshold (Ct) values were 95% removal of caffeine, and absence of viral copies reflect the efficiency of the treatment plants in Chennai city. Among the detected OTs, a combination of maximum dynamic range and high concurrence percentage was observed for caffeine and N1 gene of SARS-CoV-2. Hence, we suggest that caffeine can be used as an indicator for the removal of SARS-CoV-2 by STPs. Our predicted estimated number of cases are in line with the available clinical data from the catchments. Densely distributed population of the Koyambedu catchment could be partly responsible for the high proportion of estimated infected individuals during the study period., Graphical abstract Unlabelled Image

The impacts of the ongoing coronavirus pandemic highlight the importance of environmental monitoring to inform public health safety. Wastewater based epidemiology (WBE) has drawn interest as a tool for analysis of biomarkers in wastewater networks. Wide scale implementation of WBE requires a variety of field deployable analytical tools for real-time monitoring. Nanobiotechnology enabled sensing platforms offer potential as biosensors capable of highly efficient and sensitive detection of target analytes. This review provides an overview of the design and working principles of nanobiotechnology enabled biosensors and recent progress on the use of biosensors in detection of biomarkers. In addition, applications of biosensors for analysis of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus are highlighted as they relate to the potential expanded use of biosensors for WBE-based monitoring. Finally, we discuss the opportunities and challenges in future applications of biosensors in WBE for effective monitoring and investigation of public health threats., Graphical abstract Image 1

There is a need for a simple water sampling technique to enable routine monitoring of community drug consumption through wastewater-based epidemiology (WBE). This study investigates the potential use of diffusive gradients in thin films to sample organic compounds (o-DGT) for WBE. Three types of resin gels (HLB, XAD 18, and XDA-1) within o-DGT samplers each were deployed in triplicate at the inlets of two sewage treatment plants of Southern Asian cities. The target compounds included 15 illicit drugs and 18 antibiotics. A comprehensive evaluation was undertaken regarding each resin's ability to accumulate the target compounds and accuracy by comparing active samples. The organic compounds accumulated on each resin gel were characterised at the molecular level using Fourier transform ion-cyclotron-resonance mass spectrometry (FT-ICR MS). The results showed that the HLB resin performed better than the XAD 18 and XDA-1 resins. Based on calculations using the HLB-DGT data, methamphetamine and heroin were the two most popular illicit drugs consumed among the studied populations, and were followed by ketamine and codeine, which agreed well with the authoritative reports and reference data. The total drug consumption in Hanoi was one order of magnitude higher than that in Guangzhou, thus implying a probably more serious drug situation in the former. Overall, the findings of this study demonstrate that o-DGT passive samplers are a promising tool for WBE studies, particularly at WWTPs or in urban streams where an automatic sampler for taking composite water samples is absent.

The inclusion of new psychoactive substances (NPS) in the wastewater-based epidemiology approach presents challenges, such as the reduced number of users that translates into low concentrations of residues and the limited pharmacokinetics information available, which renders the choice of target biomarker difficult. The sampling during special social settings, the analysis with improved analytical techniques, and data processing with specific workflow to narrow the search, are required approaches for a successful monitoring. This work presents the application of a qualitative screening technique to wastewater samples collected during a city festival, where likely users of recreational substances gather and consequently higher residual concentrations of used NPS are expected. The analysis was performed using liquid chromatography coupled to high-resolution mass spectrometry. Data were processed using an algorithm that involves the extraction of accurate masses (calculated based on molecular formula) of expected m/z from an in-house database containing about 2;000 entries, including NPS and transformation products. We positively identified eight NPS belonging to the classes of synthetic cathinones, phenethylamines and opioids. In addition, the presence of benzodiazepine analogues, classical drugs and other licit substances with potential for abuse was confirmed. The screening workflow based on a database search was useful in the identification of NPS biomarkers in wastewater. The findings highlight the specific classical drugs and low NPS use in the Netherlands. Additionally, meta-chlorophenylpiperazine (mCPP), 2,5-dimethoxy-4-bromophenethylamine (2C-B), and 4-fluoroamphetamine (FA) were identified in wastewater for the first time. (C) 2017 Elsevier Ltd. All rights reserved.

Human biomonitoring, i.e. the determination of chemicals and/or their metabolites in human specimens, is the most common and potent tool for assessing human exposure to pesticides, but it suffers from limitations such as high costs and biases in sampling. Wastewater-based epidemiology (WBE) is an innovative approach based on the chemical analysis of specific human metabolic excretion products (biomarkers) in wastewater, and provides objective and real-time information on xenobiotics directly or indirectly ingested by a population. This study applied the WBE approach for the first time to evaluate human exposure to pesticides in eight cities across Europe. 24 h-composite wastewater samples were collected from the main wastewater treatment plants and analyzed for urinary metabolites of three classes of pesticides, namely triazines, organophosphates and pyrethroids, by liquid chromatography-tandem mass spectrometry. The mass loads (mg/day/1000 inhabitants) were highest for organophosphates and lowest for triazines. Different patterns were observed among the cities and for the various classes of pesticides. Population weighted loads of specific biomarkers indicated higher exposure in Castellon, Milan, Copenhagen and Bristol for pyrethroids, and in Castellon, Bristol and Zurich for organophosphates. The lowest mass loads (mg/day/1000 inhabitants) were found in Utrecht and Oslo. These results were in agreement with several national statistics related to pesticides exposure such as pesticides sales. The daily intake of pyrethroids was estimated in each city and it was found to exceed the acceptable daily intake (ADI) only in one city (Castellon, Spain). This was the first large-scale application of WBE to monitor population exposure to pesticides. The results indicated that WBE can give new information about the "average exposure" of the population to pesticides, and is a useful complementary biomonitoring tool to study population-wide exposure to pesticides. (C) 2017 Elsevier Ltd. All rights reserved.

Tobacco smoking remains an important public health issue worldwide. Assessment of exposure to tobacco-related toxicants and carcinogens at the population level is thus an essential population health indicator. This can be achieved by wastewater-based epidemiology (WBE), which relies on the analysis of biomarkers in wastewater. However, required analytical methods for the simultaneous measurement of tobacco-related toxicants and carcinogens in wastewater are not available. In this study, a new analytical procedure was developed and validated to measure tobacco-related alkaloids, carcinogens, and their metabolites in raw wastewater, including anabasine (ANABA), anatabine (ANATA), cotinine (COT), trans-3'-hydroxycotinine (COT-OH), N-nitro-soanabasine (NAB), N-nitrosoanatabine (NAT), N-nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and 4-(methylnitrosamino)-1-(3-pyridy1)-1-butanol (NNAL), NNAL-N-beta-glucuronide, and NNAL-O-beta-glucuronide. Different parameters were optimized for the solid-phase extraction procedure and instrumental analysis using liquid chromatography tandem mass spectrometry. The optimized method was fully validated, resulting in acceptable within-run and between-run precision (

There is limited knowledge on the global prescription and consumption patterns of therapeutic (TD) and illicit drugs (ID). Pooled urine analysis and wastewater-based epidemiology (WBE) has been used for local-based drug screening. It is, however, difficult to study the global epidemiology due to difficulties in obtaining samples. The aims of the study were to test the detectability of TD and ID in airplane wastewater samples categorized according to their geographical origin. Wastewater samples (n= 17) were collected from long-distance flights and prepared with enzymatic conjugate cleaving followed by either precipitation or solid phase extraction. Aliquots were analysed on various liquid chromatography – mass spectrometers. TDs were grouped according to their Anatomical Therapeutic Chemical (ATC) codes. Identification confidence was assigned to three levels based on variables including detection on multiple instruments and number of targets per compound. A total of 424 compounds were identified across all samples, distributed on 87 unique TD and 2 ID. Two principal components in a principal component analysis separated three clusters of wastewater samples corresponding to geographical origin of the airplanes with therapeutic subgroup ATC codes as variables. Airplane wastewater analysis is useful for identifying targets for WBE and toxicological analysis and explore drug use and abuse patterns.

Wastewater-based epidemiology is an increasingly popular method for analysing drugs or metabolites excreted by populations. The in-sewer transformation of biomarkers is important but often receives little consideration in published studies. Many studies publish stability under biofilm-free conditions only, which do not represent actual sewer conditions. This study aims to fill a gap in the field by comparing the wastewater stability of 33 licit drug and pharmaceutical biomarkers in biofilm-free (BFF) conditions to stability in sewer biofilm reactors. All but one biomarker was stable under BFF conditions, whereas most transformed in sewer biofilm reactors. Sewer reactor results tended to overestimate the degradation in pilot and actual sewers, whereas BFF stability had no clear relationship to stability in pilot and actual sewers. Our results provide additional basis for more informed interpretation of biofilm-free and sewer reactor stability results for past and future WBE studies.

Methamphetamine has become one of the most widely used illicit drugs in China. To understand the current situation in China, the prevalence and consumption of methamphetamine were estimated through wastewater-based epidemiology (WBE) in the present study. Methamphetamine concentrations ranged from 42.6 ng/L (Harbin) to 700 ng/L (Xi’an) in influent wastewater samples collected from 27 wastewater treatment plants (WWTPs) in 22 Chinese cities. The estimated consumption of methamphetamine was 23.0 (Dingxi) to 376 (Xi’an) mg/day/1000 inhabitants with a mean value of 157 mg/day/1000 inhabitants. The annual consumption in 2018 was estimated to be 84 tons (95% confidence interval, 44–136), which was 26% lower than that in 2014. The prevalence of methamphetamine use was 0.64% (95% confidence interval, 0.18–1.25), indicating that more than five million people used methamphetamine in 2018. Although drug abuse is common in the country, the consumption showed a different spatial pattern, with the highest values in Central China and the lowest use in Northeast China, so drug use is still considered a geographic and culture-dependent behaviour. The results indicated that WBE can not only be used to assess the trends of illicit drug use, but also to analyse the spatial differences in the whole country, which will provide complementary evidence for the prevention and control of methamphetamine use.

An individual’s excreta contains chemical information that reflects the chemicals one has consumed or been exposed to. Wastewater-based epidemiology (WBE) is the discipline concerned with mining the chemical information from municipal wastewater. WBE, also known as wastewater epidemiology, sewer epidemiology, urban water fingerprinting or sewage chemical information monitoring (SCIM), has been applied in populations around the globe to mainly measure chemical consumption and exposure patterns. In particular, WBE studies have added to our knowledge of illicit and licit drug consumption patterns, and shows increasing potential as a tool for measuring aspects of public health and socioeconomics. This chapter presents readers with an overview of key methodologies, advances and perspectives in adapting WBE as a tool for better understanding relationships between biochemical consumption and exposure behaviour with public health and socioeconomics.

AIMS: To perform a wastewater-based analysis to explore the impact of newly legalized retail cannabis sales on its use and to determine if this approach could estimate the size of the legal market place, which began 1 August 2014 in the study area. DESIGN: Raw wastewater samples were collected and analyzed over the 3-year period from 2014 – 2016. SETTING AND PARTICIPANTS: Samples obtained from the two wastewater treatment plants that serviced a municipality of 200,000 people in Washington State, USA. MEASUREMENTS: Quantitative analysis of 24-hour composite influent samples for the metabolite of the active ingredient in cannabis, 11-nor-9-Carboxy-Δ9-tetrahydrocannabinol (THC-COOH) were performed by liquid chromatography coupled to mass spectrometry. FINDINGS: Wastewater was used for the first time to monitor a drug regulatory change showing that sales of legal cannabis increased at a rate of nearly 70% per quarter, over the same 3-year period estimates for THC-COOH in the wastewater increased by 9% per quarter, resulting in a doubling in consumption over this time period. Estimating the proportion of the total cannabis market supplied by State-regulated cannabis is not currently achievable. CONCLUSION: A wastewater derived measure of cannabis consumption suggests a significant increase in consumption and that legal sales likely displaced a large portion of the illicit market. It is not possible with wastewater data to determine the degree to which this increase is due to higher purity products, more users, and/or more frequent use, despite these limitations, this data source provides important additional information for evaluating the impact of cannabis legalization.

Pollution by heavy metals and metalloids is detrimental to human health due to their toxic, genotoxic, and carcinogenic effects. The traditional approach to assess the extent of environmental and occupational exposures of metals is human biomonitoring (HBM). This method has several limitations, including invasiveness, sampling bias, cost- and time-intensiveness, and ethical issues. This suggests the need for a more robust, non-invasive, epidemiological tool for assessment of exposure to metals and their public health effects. Recently, wastewater-based epidemiology (WBE) has been suggested and utilized as a novel approach to accurately determine the extent of exposure to multiple substances on the population level. We suggest the potential application of WBE to the study of metal exposure on the population level, including possible biomarkers for wastewater analysis of 10 metals belonging to three categories according to health effects and nutritional benefits, and its public health implications. Similar to previous studies of exposure to regulated or illegal drugs, unregulated legal substances, and pesticides, WBE can be applied to the study of metal exposure in a given community. Parental substance biomarkers (PSBs), metabolic substance biomarkers (MSBs), and non-substance biomarkers (NSBs) of 10 common metals are available for consideration in wastewater analysis. The use of WBE would allow for the interpretation of the relationship between metal exposure and population health, reveal synergistic effects of different health factors, and model public health risks under different scenarios.

Anabolic-androgenic steroids are synthetic compounds prohibited due to their performance-enhancing characteristics. The use of these substances is known to cause health-related issues, which highlights the importance of being able to evaluate the scale of consumption by the general population. However, most available research on the analysis of anabolic steroids is focused on animals and athletes in connection with doping. The potential of wastewater-based epidemiology as an intelligence tool for the assessment of community level use of anabolic steroids is presented herein. A liquid chromatography tandem mass spectrometry method was developed for the analysis of 10 anabolic-androgenic steroids and 14 endogenous hormones in influent wastewater. The validated method was applied to sixteen 24-hour composite wastewater influent samples that were collected over a period of five years from two wastewater treatment plants in Queensland, Australia. Nine investigated compounds were found to be present at concentrations between 14 and 611 ng L-1 which translated into 3-104 mg excreted per 1000 individuals per day. It was concluded that the developed analytical method is suitable for the analysis of AAS in wastewater matrix. Additionally, both the inclusion of metabolites and further investigation into deconjugation by enzymatic hydrolysis would aid in understanding and evaluating community anabolic steroid use. For the first time, this study presents the application of wastewater-based epidemiology on anabolic-androgenic steroids in Australia.

Mycotoxins are among the compounds of most concern for human health because of their common presence in food and their proven toxicity for human health. Human biomonitoring (HBM) studies, foodstuff analysis and dietary surveys are usually used to assess human exposure, but they are costly and time-consuming. Wastewater-based epidemiology (WBE) can complement the established exposure monitoring tools. The aim of this study was to develop a new WBE application for assessing human exposure to mycotoxins by measuring specific biomarkers in raw wastewater. Eleven substances were analysed by liquid chromatography-tandem mass spectrometry in wastewater from four cities in Italy and Spain. Only deoxynivalenol (DON) and fumonisines were found, DON being the most abundant, in accordance with HBM studies. DON intake, back-calculated by WBE, was lower than the established Health-Based Guidance values, indicating moderate exposure in the population, with no risks for human health. WBE intake estimates were very close to those reported in HBM studies, validating WBE for estimating DON intake in the population. This study describes a promising WBE approach to complement HBM studies and assess the human intake of specific classes of mycotoxins, thus helping to identify risks for human health.

New psychoactive substances (NPSs) are compounds that produce similar effects to those induced by illicit drugs (IDs), such as cocaine, cannabis, and amphetamines, but are not strictly regulated by international conventions. The consumption of NPSs is a growing public health problem in many communities. However, there is little knowledge regarding the extent and actual use of these new substances. Monitoring NPS use is arduous, and therefore, different sources of information need to be used to get more insight of the prevalence and diffusion of NPS use. Analysis of pooled urine and wastewater (WW) shows strong potential, giving a different and complementary light on this issue, although presents some limitations and challenges that must be taken into account. Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) is one of the most powerful approaches for screening a large number of NPSs because of the accurate-mass full-spectrum acquisition measurements. By using a comprehensive and updated NPS database, LC-HRMS is flexible enough to confront the ever-changing NPS market. In this “current opinion”, we give our point of view on the usefulness of pooled urine and WW analysis and on the potential application of wastewater-based epidemiology as source of information for NPS use, explaining the main bottlenecks and future perspectives in this emerging research field.

Conceptual sewer models are useful tools to assess the fate of micropollutants (MPs) in integrated wastewater systems. However, the definition of their model structure is highly subjective, and obtaining a realistic simulation of the in-sewer hydraulic retention time (HRT) is a major challenge without detailed hydrodynamic information or with limited measurements from the sewer network. This study presents an objective approach for defining the structure of conceptual sewer models in view of modelling MP fate in large urban catchments. The proposed approach relies on GIS-based information and a Gaussian mixture model to identify the model optimal structure, providing a multi-catchment conceptual model that accounts for HRT variability across urban catchment. This approach was tested in a catchment located in a highly urbanized Italian city and it was compared against a traditional single-catchment conceptual model (using a single average HRT) for the fate assessment of reactive MPs. Results showed that the multi-catchment model allows for a successful simulation of dry weather flow patterns and for an improved simulation of MP fate compared to the classical single-catchment model. Specifically, results suggested that a multi-catchment model should be preferred for (i) degradable MPs with half-life lower than the average HRT of the catchment and (ii) MPs undergoing formation from other compounds (e.g. human metabolites); or (iii) assessing MP loads entering the wastewater treatment plant from point sources, depending on their location in the catchment. Overall, the proposed approach is expected to ease the building of conceptual sewer models, allowing to properly account for HRT distribution and consequently improving MP fate estimation.

DEET (N, N-diethyl-m-toluamide) is one of the most frequently detected trace organic contaminants (TOrC) in wastewaters and is used primarily as an insect repellent. It was introduced for use in the general public in 1957. It is ubiquitously present in the environment and DEET concentrations are usually among the highest reported for TOrCs. Due to recent concerns about possible analytical interferences in detection methods being reported, this study focused on possible artifacts caused by seasonal, spatial, and diurnal variations in wastewater influent concentration of DEET. We also compared usage data to observed wastewater concentrations of seven wastewater treatment plants (WWTPs) in four different regions in the US monitored from November 2014 to November 2016. Consumption data obtained reveal patterns of consumption according to climatic regions and season. During the summer DEET usage accounts for almost 60% of all usage during a year, while during the winter months DEET usage accounts for

The present review discusses the value of wastewater-based epidemiology (WBE) to estimate alcohol consumption in a given population. It gives an overview of the works conducted in this respect to date and the comparability of the WBE-derived data with official estimates provided by the World Health Organization, generated after surveys and alcohol sales data. Finally, it provides insights into the main advantages and limitations of the WBE approach over traditional methods and aspects/knowledge to be improved/investigated.

This is the postprint (accepted manuscript) version of the article published by Taylor & Francis in the International Journal of Environmental Analytical Chemistry on December, 10th 2018. Available on-line: http://www.tandfonline.com//10.1080/03067319.2018.1554743 The levels of cocaine (COC) and its metabolites benzoylecgonine (BE) and cocaethylene (COE) in wastewater samples from Brasilia, Brazil, were determined using liquid chromatography coupled to hybrid quadrupole-time-of-flight mass spectrometry. The former metabolite was used to estimate cocaine consumption using a country-tuned correction factor that considers BE excretion rates for different routes of administration as well as the fraction of crack users in Brazil. A day-to-day analysis during a week indicates higher cocaine use in weekends with peaks on Sunday in both southern (7385 ± 121 mg day−1 1000 inhab−1) and northern (3566 ± 171 mg day−1 1000 inhab−1) areas of Brasilia. A significant high cocaine use was also observed in the Carnival Day of 2018 (6229 ± 219 mg day−1 1000 inhab−1). COE/BE ratios were used to assess COC and alcohol co-consumption since COE is produced during the co-consumption of both substances. Higher ratios were also observed during the weekend. However, higher ratios on Saturdays rather than Sundays may be explained by the decrease of BE excretion during the co-consumption with alcohol, which may lead to an underestimation of the cocaine use estimates on Saturdays, as well as by different patterns of co-consumption by powder and crack users, where the latter usually drink lower amounts of alcohol. Our data suggest that only 3% of cocaine was seized by the local Technical Police during the study This work was supported by the Federal District Research Foundation under Grant [193.000.916/ 2015]; the Spanish Agencia Estatal de Investigación under Grant [CTM2017-84763-C3-2-R]; the Galician Council of Culture, Education and Universities under Grants [ED431C2017/36]; IGM postdoctoral contract, Plan Galego I2C-Modalidade B, ED481D 2017/003; and FEDER/ERDF SI

SARS-CoV-2 pandemic is having a devastating effect on human lives. Recent reports have shown that majority of the individuals recovered from COVID-19 have serious health complications, which is going to be a huge economic burden globally. Given the wide-spread transmission of SARS-CoV-2 it is almost impossible to test every individual in densely populated countries. Recent reports have shown that sewage-based surveillance can be used as holistic approach to understand the spread of the pandemic within a population or area. Here we have estimated the spread of SARS-CoV-2 in the city of Hyderabad, India, which is a home for nearly 10 million people. The sewage samples were collected from all the major sewage treatment plants (STPs) and were processed for detecting the viral genome using the standard Reverse Transcription Polymerase Chain Reaction (RT-PCR) method. Interestingly, inlet samples of STPs were positive for SARS-CoV-2, while the outlets were negative, which indicates that the standard sewage treatment methods are efficient in eliminating the SARS-CoV-2 viral particles. Based on the detected viral gene copies per litre and viral particle shedding per individual, the total number of individuals exposed to SARS-CoV-2 was estimated. Through this study we suggest that sewage-based surveillance is an effective approach to study the infection dynamics, which helps in efficient management of the SARS-CoV-2 spread., Graphical abstract Unlabelled Image, Highlights • Study reports the least possible estimate of SARS-CoV-2 infected. • This study estimates the infected and actively spreading population in Hyderabad, India during the study time course • Our study is one of the first comprehensive report of wastewater-based SARS-CoV-2 surveillance in India • The effluent from STPs were negative to SARS-CoV-2 indicating the efficient treatment

Wastewater-based epidemiology (WBE) can be a useful tool to face some of the existing challenges in monitoring the use of new psychoactive substances (NPS), as it can provide objective and updated information. This Europe-wide study aimed to verify the suitability of WBE for investigating the use of NPS. Selected NPS were monitored in urban wastewater by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). The main classical illicit drugs were monitored in the same samples to compare their levels with those of NPS. Raw composite wastewater samples were collected in 2016 and 2017 in 14 European countries (22 cities) following best practice sampling protocols. Methcathinone was most frequent (>65% of the cities), followed by mephedrone (>25% of the cities), and only mephedrone, methcathinone and methylone were found in both years. This study depicts the use of NPS in Europe, confirming that it is much lower than the use of classical drugs. WBE proved able to assess the qualitative and quantitative spatial and temporal profiles of NPS use. The results show the changeable nature of the NPS market and the importance of large WBE monitoring campaigns for selected priority NPS. WBE is valuable for complementing epidemiological studies to follow rapidly changing profiles of use of drugs. Supplementary material: [https://cherry.chem.bg.ac.rs/handle/123456789/4396]

Infectious diseases caused by pathogenic microbes have posed a major health issue for the public, such as the ongoing COVID-19 global pandemic. In recent years, wastewater-based epidemiology (WBE) is emerging as an effective and unbiased method for monitoring public health. Despite its increasing importance, the advancement of WBE requires more competent and streamlined analytical platforms. Herein we discuss the interactions between WBE and droplet microfluidics, focusing on the analysis of pathogens in droplets, which is hard to be tackled by traditional analytical tools. We highlight research works from three aspects, namely, quantitation of pathogen biomarkers in droplets, single-cell analysis in droplets, and living cell biosensors in droplets, as well as providing future perspectives on the synergy between WBE and droplet microfluidics.

While the extent of pharmaceutical consumption within a society/community is of high relevance to its health, economy and general wellbeing, this data is often not readily available. Herein, we strengthen a wastewater–based epidemiology (WBE) approach as a way to track the consumption of pharmaceuticals within the sampled community. This method is less laborious than established questionnaire or databases approaches and allows a higher temporal and spatial resolution. The WBE approach was conducted by sampling influent wastewater from two wastewater treatment plants of different size. A total of 39 targeted compounds were quantified by liquid chromatography coupled with tandem mass spectrometry. The number of prescriptions and the defined daily doses for each prescription was obtained from the reference database of The Catalan Health System to validate the wastewater-based approach. The wastewater sampling and the data inquiry were both executed during the same period (October 2019) and standardised for comparison to treatments per 1,000 inhabitants per day. The back-calculation parameters were improved from previous studies by including the faecal excretion rate of the pharmaceuticals. For prescription only pharmaceuticals, where prescription numbers are expected to be a good estimate of consumption, our WBE approach agreed with 27 out of 32 (

Wastewater based epidemiology (WBE) has emerged as a reliable strategy to assess the coronavirus disease 2019 (COVID-19) pandemic. Recent publications suggest that SARS-CoV-2 detection in wastewater is technically feasible; however, many different protocols are available and most of the methods applied have not been properly validated. To this end, different procedures to concentrate and extract inactivated SARS-CoV-2 and surrogates were initially evaluated. Urban wastewater seeded with gamma-irradiated SARS-CoV-2, porcine epidemic diarrhea virus (PEDV), and mengovirus (MgV) was used to test the concentration efficiency of an aluminum-based adsorption-precipitation method and a polyethylene glycol (PEG) precipitation protocol. Moreover, two different RNA extraction methods were compared in this study: a commercial manual spin column centrifugation kit and an automated protocol based on magnetic silica beads. Overall, the evaluated concentration methods did not impact the recovery of gamma-irradiated SARS-CoV-2 nor MgV, while extraction methods showed significant differences for PEDV. Mean recovery rates of 42.9 ± 9.5%, 27.5 ± 14.3% and 9.0 ± 2.2% were obtained for gamma-irradiated SARS-CoV-2, PEDV and MgV, respectively. Limits of detection (LoD95%) for five genomic SARS-CoV-2 targets (N1, N2, gene E, IP2 and IP4) ranged from 1.56 log genome equivalents (ge)/mL (N1) to 2.22 log ge/mL (IP4) when automated system was used; while values ranging between 2.08 (N1) and 2.34 (E) log ge/mL were observed when using column-based extraction method. Different targets were also evaluated in naturally contaminated wastewater samples with 91.2%, 85.3%, 70.6%, 79.4% and 73.5% positivity, for N1, N2, E, IP2 and IP4, respectively. Our benchmarked comparison study suggests that the aluminum precipitation method coupled with the automated nucleic extraction represents a method of acceptable sensitivity to provide readily results of interest for SARS-CoV-2 WBE surveillance., The study was funded by grants from CSIC (202070E101), Generalitat Valenciana (Covid_19-SCI), MICINN co-founded by AEI/FEDER, UE (AGL2017-82909), and MICINN/AEI (PID2019-105509RJ-I00). EC-F is recipient of a predoctoral contract from the MICINN, Call 2018. WR is holder of the APOSTD/2018/150 postdoctoral contract from Generalitat Valenciana.