Monitoring genetic population biomarkers for wastewater-based epidemiology

We report a rapid “sample-to-answer” platform that can be used for the quantitative monitoring of genetic biomarkers within communities through the analysis of wastewater. The assay is based on the loop-mediated isothermal amplification (LAMP) of nucleic acid biomarkers and shows for the first time the ability to rapidly quantify human-specific mitochondrial DNA (mtDNA) from raw untreated wastewater samples. mtDNA provides a model population biomarker associated with carcinogenesis including breast, renal and gastric cancers. To enable a sample-to-answer, field-based technology, we integrated a filter to remove solid impurities and perform DNA extraction and enrichment into a low cost lateral flow-based test. We demonstrated mtDNA detection over seven consecutive days, achieving a limit of detection of 40 copies of human genomic DNA per reaction volume. The assay can be performed at the site of sample collection, with minimal user intervention, yielding results within 45 min and providing a method to monitor public health from wastewater

Community sewage sensors towards evaluation of drug use trends: detection of cocaine in wastewater with DNA-directed immobilization aptamer sensors

Illicit drug use has a global concern and effective monitoring and interventions are highly required to combat drug abuse. Wastewater-based epidemiology (WBE) is an innovative and cost-effective approach to evaluate community-wide drug use trends, compared to traditional population surveys. Here we report for the first time, a novel quantitative community sewage sensor (namely DNA-directed immobilization of aptamer sensors, DDIAS) for rapid and cost-effective estimation of cocaine use trends via WBE. Thiolated single-stranded DNA (ssDNA) probe was hybridized with aptamer ssDNA in solution, followed by co-immobilization with 6-mercapto-hexane onto the gold electrodes to control the surface density to effectively bind with cocaine. DDIAS was optimized to detect cocaine at as low as 10 nM with a dynamic range from 10 nM to 5 μM, which were further employed for the quantification of cocaine in wastewater samples collected from a wastewater treatment plant in seven consecutive days. The concentration pattern of the sampling week is comparable with that from mass spectrometry. Our results demonstrate that the developed DDIAS can be used as community sewage sensors for rapid and cost-effective evaluation of drug use trends, and potentially implemented as a powerful tool for on-site and real-time monitoring of wastewater by un-skilled personnel

Analytical Profiling of Airplane Wastewater - a New Matrix for Mapping Worldwide Patterns of Drug Use and Abuse

Abstract 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.</jats:p

VIRAL PARTICLE DETECTION IN WASTEWATER

Wastewater is an underutilized resource for recognizing the epidemiology of pathogen transmission and infections within a community that has the potential to mitigate disease outbreaks. Pathogens of public health concern, such as adenovirus, norovirus, and SARS-CoV-2, occur in wastewater from excretion of infected individuals. Despite the potential for wastewater pathogen data to predict future outbreaks, the relationship between viral shedding patterns in wastewater and case counts of infectious disease remains unclear. An evaluation of the peer-reviewed literature was conducted to compile characteristics of adenovirus, norovirus, and SARS-CoV-2 in the wastewater environment. These characteristics were considered in prioritizing the key pathogens for wastewater monitoring programs. SARS-CoV-2 is recommended to have the highest priority due to the recent pandemic, in addition to high caseload, shedding volumes, and contagiousness. Adenovirus receives the next highest priority because the positive PCR detections have been increasing (indicating there may be an increase in cases), the large case burden, and pathogen hardiness in wastewater. Norovirus receives the final ranking of the three but should still be a priority due to high caseload, shedding volume, and contagiousness. Wastewater-based epidemiology is gaining recognition as a substantial sector of epidemiology and public health research. This work provides a criteria platform to inform future quantitative microbial risk assessments that will build a framework for interpreting wastewater pathogen monitoring data. This framework will aid community leaders in public health decision-making based on the occurrence of specific pathogens detected in sewage

Detection of influenza virus in urban wastewater during the season 2022/2023 in Sicily, Italy

Introduction: Seasonal influenza generally represents an underestimated public health problem with significant socioeconomic implications. Monitoring and detecting influenza epidemics are important tasks that require integrated strategies. Wastewater-based epidemiology (WBE) is an emerging field that uses wastewater data to monitor the spread of disease and assess the health of a community. It can represent an integrative surveillance tool for better understanding the epidemiology of influenza and prevention strategies in public health. Methods: We conducted a study that detected the presence of Influenza virus RNA using a wastewater-based approach. Samples were collected from five wastewater treatment plants in five different municipalities, serving a cumulative population of 555,673 Sicilian inhabitants in Italy. We used the RT-qPCR test to compare the combined weekly average of Influenza A and B viral RNA in wastewater samples with the average weekly incidence of Influenza-like illness (ILI) obtained from the Italian national Influenza surveillance system. We also compared the number of positive Influenza swabs with the viral RNA loads detected from wastewater. Our study investigated 189 wastewater samples. Results: Cumulative ILI cases substantially overlapped with the Influenza RNA load from wastewater samples. Influenza viral RNA trends in wastewater samples were similar to the rise of ILI cases in the population. Therefore, wastewater surveillance confirmed the co-circulation of Influenza A and B viruses during the season 2022/2023, with a similar trend to that reported for the weekly clinically confirmed cases. Conclusion: Wastewater-based epidemiology does not replace traditional epidemiological surveillance methods, such as laboratory testing of samples from infected individuals. However, it can be a valuable complement to obtaining additional information on the incidence of influenza in the population and preventing its spread

Making waves : wastewater-based epidemiology for COVID-19 - approaches and challenges for surveillance and prediction

The presence of SARS-CoV-2 in the feces of infected patients and wastewater has drawn attention, not only to the possibility of fecal-oral transmission but also to the use of wastewater as an epidemiological tool. The COVID-19 pandemic has highlighted problems in evaluating the epidemiological scope of the disease using classical surveillance approaches, due to a lack of diagnostic capacity, and their application to only a small proportion of the population. As in previous pandemics, statistics, particularly the proportion of the population infected, are believed to be widely underestimated. Furthermore, analysis of only clinical samples cannot predict outbreaks in a timely manner or easily capture asymptomatic carriers. Threfore, community-scale surveillance, including wastewater-based epidemiology, can bridge the broader community and the clinic, becoming a valuable indirect epidemiological prediction tool for SARS-CoV-2 and other pandemic viruses. This article summarizes current knowledge and discusses the critical factors for implementing wastewater-based epidemiology of COVID-19

Wastewater-based drug epidemiology to estimate societal drug use: A critical review

Illicit drug use has many consequences resulting in social, health, and economic harm. An objective method of quantifying societal drug use would be useful for the efficient directing of the efforts of law enforcement, medical facilities and policy makers, and to inform the community. To this end, efforts to ascertain societal drug use have relied upon community surveys and extrapolation from law enforcement seizures, which often present the data as biased or skewed due to a small sample size and other associated limitations. In recent times, wastewater-based drug epidemiology (WBDE) has been proposed as a suitable means to objectively quantify societal drug use. WBDE is the study of the incidence and distribution of drug use within a population and the its factors affecting the health and welfare. It is a method contingent upon the concept of measuring drug metabolites or biomarkers in wastewater (WW), from which levels of societal drug use are estimated and quantified through extrapolation and back-calculations. The aim of this study was to critically review the various methods of WBDE that have been applied in Australia and in Europe. The outcomes of the assessment pertaining to their validity in directly and objectively measuring societal drug use will be presented. Keywords: wastewater-based drug epidemiology, wastewater-based epidemiology, illicit drugs, drug abuse, estimate, societal drug use, populatio

A Dynamic Quantitative Microbial Risk Assessment for Norovirus in Potable Reuse System

This study describes the results of a dynamic quantitative microbial risk assessment (QMRA) for norovirus (NoV) that was used to evaluate the relative significance of foodborne, person-to-person, and person-to-sewage-to-person transmission pathways. This last pathway was incorporated into simulated potable reuse systems to evaluate the adequacy of typical treatment trains, operational conditions, and regulatory frameworks. The results confirm that secondary and foodborne transmission dominate the overall risk calculation and that waterborne NoV likely contributes no appreciable public health risk, at least in the scenarios modeled in this study. De facto reuse with an environmental buffer storage time of at least 30 days was comparable or even superior to direct potable reuse (DPR) when compound failures during advanced treatment were considered in the model. Except during these low-probability failure events, DPR generally remained below the 10−4 annual risk benchmark for drinking water. Based on system feedback and the time-dependent pathogen load to the community\u27s raw sewage, this model estimated median raw wastewater NoV concentrations of 107–108 genome copies per liter (gc/L), which is consistent with high-end estimates in recent literature

Multi-year interlaboratory exercises for the analysis of illicit drugs and metabolites in wastewater:development of a quality control system

Thirty-seven laboratories from 25 countries present the development of an inter-laboratory testing scheme for the analysis of seven illicit drug residues in standard solutions, tap- and wastewater. Almost 10 000 concentration values were evaluated: triplicates of up to five samples and 26 laboratories per year. The setup was substantially improved with experiences gained across the six repetitions (e.g. matrix type, sample conditions, spiking levels). From this, (pre-)analytical issues (e.g. pH adjustment, filtration) were revealed for specific analytes which resulted in formulation of best-practice protocols for inter-laboratory setup and analytical procedures. The results illustrate the effectiveness of the inter-laboratory setup to assess laboratory performance in the framework of wastewater-based epidemiology. The exercise proved that measurements of laboratories were of high quality (>80% satisfactory results for six out of seven analytes) and that analytical follow-up is important to assist laboratories in improving robustness of wastewater-based epidemiology results