Identifying potential bacterial pathogens in wastewater by expert reliability analysis of full-length 16S rRNA and untargeted RNA sequencing data processed with four bioinformatic workflows.

Municipal wastewater may serve as a critical community-composite sample for monitoring bacteria excreted by the contributing population, providing insights into public health risks and microbial diversity. The present study emphasizes the integration of DNA (full-length 16S rRNA) methods, untargeted RNA methods, and different bioinformatic protocols to identify potential human bacterial pathogens in wastewater. Results revealed that, DNA surveillance identified roughly 50 % of the sequencing reads were associated with potentially pathogenic bacteria, as compared to RNA surveillance, which identified roughly 33 % of the reads as associated with potential bacterial pathogens.

In-depth comparison of untargeted and targeted sequencing for detecting virus diversity in wastewater.

Sequencing approaches may enable monitoring of a broad range of viruses in wastewater, including potential emerging and non-reportable human viruses. Considering the fact that metagenomic sequencing may be non-specific for low-abundance human viruses, integration of viral amplification and enrichment strategies are proposed to enhance the accurate detection of a broad range of human viruses in municipal wastewater. In this study, we focused on the in-depth comparison analysis of three untargeted amplification methods (Multiple Displace Amplification [MDA], Reverse Transcription - MDA [RT-MDA], and a PCR-based random amplification [PCR-based]) and one targeted method (Twist Comprehensive Viral Research Panel [TWIST]) for detecting virus diversity in wastewater.

Community case study: an academia-industry-government partnership that monitors and predicts outbreaks in Tri-County Detroit area since 2017.

The Tri-County Detroit Area (TCDA) is the 12th most populous metropolitan area in the United States with over three million people. Multiple communicable diseases are endemic in the TCDA. In 2017, to explore innovative methods that may provide early warnings of outbreaks affecting populations in the TCDA, an exploratory partnership that was funded by a U.

Wastewater Surveillance for Poliovirus in Selected Jurisdictions, United States, 2022-2023.

Wastewater testing can inform public health action as a component of polio outbreak response. During 2022-2023, a total of 7 US jurisdictions (5 states and 2 cities) participated in prospective or retrospective testing of wastewater for poliovirus after a paralytic polio case was identified in New York state. Two distinct vaccine-derived poliovirus type 2 viruses were detected in wastewater from New York state and New York City during 2022, representing 2 separate importation events.

Comparative analyses of SARS-CoV-2 RNA concentrations in Detroit wastewater quantified with CDC N1, N2, and SC2 assays reveal optimal target for predicting COVID-19 cases.

To monitor COVID-19 through wastewater surveillance, global researchers dedicated significant endeavors and resources to develop and implement diverse RT-qPCR or RT-ddPCR assays targeting different genes of SARS-CoV-2. Effective wastewater surveillance hinges on the appropriate selection of the most suitable assay, especially for resource-constrained regions where scant technical and socioeconomic resources restrict the options for testing with multiple assays. Further research is imperative to evaluate the existing assays through comprehensive comparative analyses.

A broad wastewater screening and clinical data surveillance for virus-related diseases in the metropolitan Detroit area in Michigan.

Background: Periodic bioinformatics-based screening of wastewater for assessing the diversity of potential human viral pathogens circulating in a given community may help to identify novel or potentially emerging infectious diseases. Any identified contigs related to novel or emerging viruses should be confirmed with targeted wastewater and clinical testing.

Results: During the COVID-19 pandemic, untreated wastewater samples were collected for a 1-year period from the Great Lakes Water Authority Wastewater Treatment Facility in Detroit, MI, USA, and viral population diversity from both centralized interceptor sites and localized neighborhood sewersheds was investigated.

Effect of wastewater collection and concentration methods on assessment of viral diversity.

Monitoring of potentially pathogenic human viruses in wastewater is of crucial importance to understand disease trends in communities, predict potential outbreaks, and boost preparedness and response by public health departments. High throughput metagenomic sequencing opens an opportunity to expand the capabilities of wastewater surveillance. However, there are major bottlenecks in the metagenomic enabled wastewater surveillance, including the complexities in selecting appropriate sampling and concentration/virus enrichment methods as well as in bioinformatic analysis of complex samples with low human virus concentrations.

Targeting a free viral fraction enhances the early alert potential of wastewater surveillance for SARS-CoV-2: a methods comparison spanning the transition between delta and omicron variants in a large urban center.

Introduction: Wastewater surveillance has proven to be a valuable approach to monitoring the spread of SARS-CoV-2, the virus that causes Coronavirus disease 2019 (COVID-19). Recognizing the benefits of wastewater surveillance as a tool to support public health in tracking SARS-CoV-2 and other respiratory pathogens, numerous wastewater virus sampling and concentration methods have been tested for appropriate applications as well as their significance for actionability by public health practices.

Methods: Here, we present a 34-week long wastewater surveillance study that covers nearly 4 million residents of the Detroit (MI, United States) metropolitan area.

Wastewater surveillance beyond COVID-19: a ranking system for communicable disease testing in the tri-county Detroit area, Michigan, USA.

Introduction: Throughout the coronavirus disease 2019 (COVID-19) pandemic, wastewater surveillance has been utilized to monitor the disease in the United States through routine national, statewide, and regional monitoring projects. A significant canon of evidence was produced showing that wastewater surveillance is a credible and effective tool for disease monitoring. Hence, the application of wastewater surveillance can extend beyond monitoring SARS-CoV-2 to encompass a diverse range of emerging diseases.

Simple methods for early warnings of COVID-19 surges: Lessons learned from 21 months of wastewater and clinical data collection in Detroit, Michigan, United States.

Wastewater-based epidemiology (WBE) has drawn great attention since the Coronavirus disease 2019 (COVID-19) pandemic, not only due to its capability to circumvent the limitations of traditional clinical surveillance, but also due to its potential to forewarn fluctuations of disease incidences in communities. One critical application of WBE is to provide "early warnings" for upcoming fluctuations of disease incidences in communities which traditional clinical testing is incapable to achieve. While intricate models have been developed to determine early warnings based on wastewater surveillance data, there is an exigent need for straightforward, rapid, broadly applicable methods for health departments and partner agencies to implement.

Surveillance of SARS-CoV-2 in nine neighborhood sewersheds in Detroit Tri-County area, United States: Assessing per capita SARS-CoV-2 estimations and COVID-19 incidence.

Wastewater-based epidemiology (WBE) has been suggested as a useful tool to predict the emergence and investigate the extent of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, we screened appropriate population biomarkers for wastewater SARS-CoV-2 normalization and compared the normalized SARS-CoV-2 values across locations with different demographic characteristics in southeastern Michigan. Wastewater samples were collected between December 2020 and October 2021 from nine neighborhood sewersheds in the Detroit Tri-County area.

Five-week warning of COVID-19 peaks prior to the Omicron surge in Detroit, Michigan using wastewater surveillance.

Wastewater-based epidemiology (WBE) is useful in predicting temporal fluctuations of COVID-19 incidence in communities and providing early warnings of pending outbreaks. To investigate the relationship between SARS-CoV-2 concentrations in wastewater and COVID-19 incidence in communities, a 12-month study between September 1, 2020, and August 31, 2021, prior to the Omicron surge, was conducted. 407 untreated wastewater samples were collected from the Great Lakes Water Authority (GLWA) in southeastern Michigan.

Minimizing errors in RT-PCR detection and quantification of SARS-CoV-2 RNA for wastewater surveillance.

Wastewater surveillance for pathogens using reverse transcription-polymerase chain reaction (RT-PCR) is an effective and resource-efficient tool for gathering community-level public health information, including the incidence of coronavirus disease-19 (COVID-19). Surveillance of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) in wastewater can potentially provide an early warning signal of COVID-19 infections in a community. The capacity of the world's environmental microbiology and virology laboratories for SARS-CoV-2 RNA characterization in wastewater is increasing rapidly.

Identification of multiple potential viral diseases in a large urban center using wastewater surveillance.

Viruses are linked to a multitude of human illnesses and can disseminate widely in urbanized environments causing global adverse impacts on communities and healthcare infrastructures. Wastewater-based epidemiology was employed using metagenomics and quantitative polymerase chain reaction (qPCR) assays to identify enteric and non-enteric viruses collected from a large urban area for potential public health monitoring and outbreak analysis. Untreated wastewater samples were collected from November 2017 to February 2018 (n = 54) to evaluate the diversity of human viral pathogens in collected samples.

Understanding temporal and spatial variations of viral disease in the US: The need for a one-health-based data collection and analysis approach.

Viral diseases exhibit spatial and temporal variation, and there are many factors that can affect their occurrence. The identification of these factors is critical in the efforts to predict and lessen viral disease burden. Because viral infection is able to spread to humans from the environment, animals, and other humans, the One-Health framework can be used to investigate the critical pathways through which viruses are transported and transmitted.

A water-focused one-health approach for early detection and prevention of viral outbreaks.

Despite consistent efforts to protect public health there is still a heavy burden of viral disease, both in the United States and abroad. In addition to conventional medical treatment, there is a need for a holistic approach for early detection and prevention of viral outbreaks at a population level. One-Health is a relatively new integrative approach to the solving of global health challenges.

Viral diversity and abundance in polluted waters in Kampala, Uganda.

Waterborne viruses are a significant cause of human disease, especially in developing countries such as Uganda. A total of 15 virus-selective samples were collected at five sites (Bugolobi Wastewater Treatment Plant (WWTP) influent and effluent, Nakivubo Channel upstream and downstream of the WWTP, and Nakivubo Swamp) in July and August 2016. Quantitative PCR and quantitative RT-PCR was performed to determine the concentrations of four human viruses (adenovirus, enterovirus, rotavirus, and hepatitis A virus) in the samples.

Diversity of DNA viruses in effluents of membrane bioreactors in Traverse City, MI (USA) and La Grande Motte (France).

This study assesses diversity of DNA viruses in the effluents of two membrane bioreactor (MBR) wastewater treatment plants (WWTPs): an MBR in the United States and an MBR in France. Viral diversity of these effluents is compared to that of a conventional activated sludge WWTP in the U.S.

Elution Is a Critical Step for Recovering Human Adenovirus 40 from Tap Water and Surface Water by Cross-Flow Ultrafiltration.

Unlabelled: This paper examines the recovery of the enteric adenovirus human adenovirus 40 (HAdV 40) by cross-flow ultrafiltration and interprets recovery values in terms of physicochemical interactions of virions during sample concentration. Prior to ultrafiltration, membranes were either blocked by exposure to calf serum (CS) or coated with a polyelectrolyte multilayer (PEM). HAdV 40 is a hydrophobic virus with a point of zero charge between pH 4.

Effect of pressure relaxation and membrane backwash on adenovirus removal in a membrane bioreactor.

Pressure relaxation and permeate backwash are two commonly used physical methods for membrane fouling mitigation in membrane bioreactor (MBR) systems. In order to assess the impact of these methods on virus removal by MBRs, experiments were conducted in a bench-scale submerged MBR treating synthetic wastewater. The membranes employed were hollow fibers with the nominal pore size of 0.

Effect of organic carbon on sorption of human adenovirus to soil particles and laboratory containers.

A key factor controlling the relationship between virus release and human exposure is how virus particles interact with soils, sediments and other solid particles in the environment and in engineered treatment systems. Finding no previous investigations of human adenovirus (HAdV) sorption, we performed a series of experiments to evaluate the role of soil organic carbon (SOC) and solution-phase dissolved organic carbon (DOC) on sorption capacity and reversibility. In preliminary methodological studies, we found that as much as 99% of HAdV was lost from inorganic buffer suspensions in polypropylene (PP) laboratory containers, but little loss occurred when using suspensions with substantial amounts of DOC or with glass containers from either type of suspension.

Leaching and ponding of viral contaminants following land application of biosolids on sandy-loam soil.

Much of the land available for application of biosolids is cropland near urban areas. Biosolids are often applied on hay or grassland during the growing season or on corn ground before planting or after harvest in the fall. In this study, mesophilic anaerobic digested (MAD) biosolids were applied at 56,000 L/ha on a sandy-loam soil over large containment lysimeters seeded to perennial covers of orchardgrass (Dactylis glomerata L.