The National Wastewater Surveillance System (NWSS): From inception to widespread coverage, 2020–2022, United States

Adams, Carly; Bias, Megan; Welsh, Rory M.; Webb, Jenna; Reese, Heather; Delgado, Stephen; Person, John; West, Rachel; Shin, Soo; Kirby, Amy

doi:10.1016/j.scitotenv.2024.171566

Cited by 16 publications

(3 citation statements)

References 23 publications

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“…Wastewater-based epidemiology (WBE), previously employed for monitoring enteric viruses like polio, has been widely applied during the COVID-19 pandemic. In 2020, the US Centers for Disease Control and Prevention (CDC) launched the National Wastewater Surveillance System (NWSS) to build and coordinate the capacity for WBE as a component of the nationwide monitoring of SARS-CoV-2 . Subsequently, groups around the world have expanded WBE to include PCR-based monitoring of known seasonal respiratory viruses including respiratory syncytial virus (RSV) and influenza A and B, and new PCR panels are expected to contribute to CDC NWSS …”

Section: Introductionmentioning

confidence: 99%

“…In 2020, the US Centers for Disease Control and Prevention (CDC) launched the National Wastewater Surveillance System (NWSS) to build and coordinate the capacity for WBE as a component of the nationwide monitoring of SARS-CoV-2. 2 Subsequently, groups around the world have expanded WBE to include PCR-based monitoring of known seasonal respiratory viruses including respiratory syncytial virus (RSV) and influenza A and B, and new PCR panels are expected to contribute to CDC NWSS. 3 Unlike PCR-based virus quantification, sequencing of viruses in wastewater has the potential to monitor many human viruses at the genome level simultaneously.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of the Impact of Concentration and Extraction Methods on the Targeted Sequencing of Human Viruses from Wastewater

Jiang,

Wang,

et al. 2024

Environ. Sci. Technol.

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Sequencing human viruses in wastewater is challenging due to their low abundance compared to the total microbial background. This study compared the impact of four virus concentration/extraction methods (Innovaprep, Nanotrap, Promega, and Solids extraction) on probe-capture enrichment for human viruses followed by sequencing. Different concentration/ extraction methods yielded distinct virus profiles. Innovaprep ultrafiltration (following solids removal) had the highest sequencing sensitivity and richness, resulting in the successful assembly of several near-complete human virus genomes. However, it was less sensitive in detecting SARS-CoV-2 by digital polymerase chain reaction (dPCR) compared to Promega and Nanotrap. Across all preparation methods, astroviruses and polyomaviruses were the most highly abundant human viruses, and SARS-CoV-2 was rare. These findings suggest that sequencing success can be increased using methods that reduce nontarget nucleic acids in the extract, though the absolute concentration of total extracted nucleic acid, as indicated by Qubit, and targeted viruses, as indicated by dPCR, may not be directly related to targeted sequencing performance. Further, using broadly targeted sequencing panels may capture viral diversity but risks losing signals for specific low-abundance viruses. Overall, this study highlights the importance of aligning wet lab and bioinformatic methods with specific goals when employing probe-capture enrichment for human virus sequencing from wastewater.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Evaluation of the Impact of Concentration and Extraction Methods on the Targeted Sequencing of Human Viruses from Wastewater

Jiang,

Wang,

et al. 2024

Environ. Sci. Technol.

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“…The signals of many other respiratory viruses have been measured in wastewater and correlated with clinical cases. 8,10,11,[13][14][15][16][17] Wastewater surveillance of respiratory-associated adenoviruses has the potential to help inform public health actions aimed at reducing transmission and inform the medical community of possible sources of respiratory illnesses.…”

Section: Graphical Abstract Introductionmentioning

confidence: 99%

Respiratory human adenovirus outbreak captured in wastewater surveillance

Kazmer,

Ammerman,

Edwards

et al. 2024

Preprint

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Adenoviruses present challenges for traditional surveillance methods since there are more than 60 types that infect humans. Wastewater-based surveillance can supplement traditional surveillance methods for gastrointestinal-associated adenoviruses, but the ability to detect trends of respiratory-associated adenoviruses in wastewater remains unclear. We quantified human adenovirus type 4 (HAdV-4) in wastewater settled solids and compared wastewater measurements to clinical cases from an outbreak investigation beginning in late September 2023. The human adenovirus type 4 target was positively correlated with clinical cases (Spearman’s rho = 0.5470, p < 0.0001) and followed a similar trend during the outbreak. We also quantified human adenovirus types 3, 7, 14, 21, 40/41, and a pan-adenovirus assay that targets all types that infect humans. The respiratory adenoviruses comprised a small fraction of the adenoviruses in wastewater and types 40/41, which typically cause gastrointestinal disease, comprised the majority of the detected adenoviruses. The efficacy of adenovirus wastewater surveillance will depend on assay specificity and the public health action available for adenovirus types.WATER IMPACT STATEMENTCommunity adenovirus surveillance is strengthened by wastewater measurements. We evaluated the correlation between wastewater measurements and clinical cases during an outbreak of respiratory adenovirus type 4 infections on a college campus. Results indicate respiratory adenoviruses comprise a small portion of the adenoviruses measured in wastewater and the utility of adenovirus wastewater surveillance depends on the type and public health actionability.GRAPHICAL ABSTRACT

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Strategy to develop and validate digital droplet PCR methods for global antimicrobial resistance wastewater surveillance

Gobbo,

Fraiture,

Van Poelvoorde

et al. 2024

Water Environment Research

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According to World Health Organization (WHO), antimicrobial resistance (AMR) is currently one of the world's top 10 health threats, causing infections to become difficult or impossible to treat, increasing the risk of disease spread, severe illness, disability, and death. Accurate surveillance is a key component in the fight against AMR. Wastewater is progressively becoming a new player in AMR surveillance, with the promise of a cost‐effective real‐time tracking of global AMR profiles in specific regions. One of the most useful analytical methods for wastewater surveillance is currently based on real‐time PCR (qPCR) and digital droplet PCR (ddPCR) technologies.As stated in the EU Wastewater Treatment Directive proposal, methodological standardization, including a workflow for method development and validation, will play a crucial role in global monitoring of AMR in wastewater. However, according to our knowledge, there are currently no qPCR and ddPCR methods for AMR surveillance available that have been validated according to international standard performance criteria. Therefore, this study proposes a workflow for the development and validation of PCR‐based methods for a harmonized and global AMR surveillance, including the construction of specific sequence databases and microbial collections for an efficient method development and method specificity evaluation. Following this strategy, we have developed and validated four duplex ddPCR methods responding to international standard performance criteria, focusing on seven AMR genes (ARG's), including extended spectrum beta‐lactam (blaCTX‐M), carbapenem (blaKPC‐2/3), tetracycline (tet(M)), erythromycin (erm(B)), vancomycin (vanA), sulfonamide (sul2), and aminoglycoside (aac(3)‐IV), as well as one indicator of antibiotic (multi‐) resistance and horizontal gene transfer, named the class I integron (intl1). The performance of these ddPCR methods was successfully assessed for their specificity, as no false‐positive and false‐negative results were observed. These ddPCR methods were also considered to be highly sensitive as showing a limit of detection below 25 copies of the targets. In addition, their applicability was confirmed using 14 wastewater samples collected from two Belgian water resource recovery facilities. The proposed study represents therefore a step forward to reinforce method harmonization in the context of the global AMR surveillance in wastewater.Practitioner Points In the context of wastewater surveillance, no PCR‐based methods for global AMR monitoring are currently validated according to international standards. Consequently, we propose a workflow to develop and validate PCR‐based methods for a harmonized and global AMR surveillance. This workflow resulted here in four duplex ddPCR methods targeting seven ARGs and one general indicator for mobilizable resistance genes. The applicability of these validated ddPCR methods was confirmed on 14 wastewater samples from two Belgian water resource recovery facilities.

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The National Wastewater Surveillance System (NWSS): From inception to widespread coverage, 2020–2022, United States

Cited by 16 publications

References 23 publications

Evaluation of the Impact of Concentration and Extraction Methods on the Targeted Sequencing of Human Viruses from Wastewater

Evaluation of the Impact of Concentration and Extraction Methods on the Targeted Sequencing of Human Viruses from Wastewater

Respiratory human adenovirus outbreak captured in wastewater surveillance

Strategy to develop and validate digital droplet PCR methods for global antimicrobial resistance wastewater surveillance

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