{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T05:07:08Z","timestamp":1790399228453,"version":"4.1.0"},"reference-count":48,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,5,4]],"date-time":"2025-05-04T00:00:00Z","timestamp":1746316800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Tuscany Region","award":["174"],"award-info":[{"award-number":["174"]}]},{"name":"Tuscany Region","award":["294"],"award-info":[{"award-number":["294"]}]},{"name":"Tuscany Region","award":["060701\/2021\/864481\/SUB\/ENV.C2"],"award-info":[{"award-number":["060701\/2021\/864481\/SUB\/ENV.C2"]}]},{"name":"Italian Government","award":["174"],"award-info":[{"award-number":["174"]}]},{"name":"Italian Government","award":["294"],"award-info":[{"award-number":["294"]}]},{"name":"Italian Government","award":["060701\/2021\/864481\/SUB\/ENV.C2"],"award-info":[{"award-number":["060701\/2021\/864481\/SUB\/ENV.C2"]}]},{"name":"EU Commission","award":["174"],"award-info":[{"award-number":["174"]}]},{"name":"EU Commission","award":["294"],"award-info":[{"award-number":["294"]}]},{"name":"EU Commission","award":["060701\/2021\/864481\/SUB\/ENV.C2"],"award-info":[{"award-number":["060701\/2021\/864481\/SUB\/ENV.C2"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Wastewater-based epidemiology (WBE) has been widely employed to track the spread of human pathogens; however, correlating wastewater data with clinical surveillance remains challenging due to population variability and environmental factors affecting wastewater composition. This study evaluated different SARS-CoV-2 normalization methods, comparing static population estimates with dynamic normalization based on common physicochemical parameters: chemical oxygen demand (COD), biochemical oxygen demand (BOD5), and ammonia (NH4-N). Wastewater samples were collected from four urban wastewater treatment plants (WWTPs) in northwestern Tuscany (Italy) from February 2021 to March 2023. The correlations between normalized viral loads and clinical COVID-19 cases were highest for static normalization (\u03c1 = 0.405), followed closely by dynamic normalization using COD and BOD5 (\u03c1 = 0.378 each). Normalization based on NH4-N was less effective. These findings suggest that chemical parameters, particularly COD and BOD5, offer a valid alternative for viral normalization when population estimates or flow rate measurements are unavailable. These parameters provide a cost-effective and practical approach for improving WBE reliability, particularly in resource-limited settings. Our results reinforce the importance of normalization in WBE to enhance its representativeness and applicability for public health surveillance.<\/jats:p>","DOI":"10.3390\/v17050672","type":"journal-article","created":{"date-parts":[[2025,5,4]],"date-time":"2025-05-04T20:42:37Z","timestamp":1746391357000},"page":"672","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Evaluating Population Normalization Methods Using Chemical Data for Wastewater-Based Epidemiology: Insights from a Site-Specific Case Study"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4078-5108","authenticated-orcid":false,"given":"Marco","family":"Verani","sequence":"first","affiliation":[{"name":"Laboratory of Hygiene and Environmental Virology, Department of Biology, University of Pisa, Via S. Zeno 35\/39, 56123 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3890-8546","authenticated-orcid":false,"given":"Ileana","family":"Federigi","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Environmental Virology, Department of Biology, University of Pisa, Via S. Zeno 35\/39, 56123 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandra","family":"Angori","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Environmental Virology, Department of Biology, University of Pisa, Via S. Zeno 35\/39, 56123 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-8106-1519","authenticated-orcid":false,"given":"Alessandra","family":"Pagani","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Environmental Virology, Department of Biology, University of Pisa, Via S. Zeno 35\/39, 56123 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Marvulli","sequence":"additional","affiliation":[{"name":"Acque S.p.A., Via A. Bellatalla 1, 56121 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudia","family":"Valentini","sequence":"additional","affiliation":[{"name":"Gaia S.p.A., Via G. Donizetti 16, 55045 Pietrasanta, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-2588-3529","authenticated-orcid":false,"given":"Nebiyu Tariku","family":"Atomsa","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Environmental Virology, Department of Biology, University of Pisa, Via S. Zeno 35\/39, 56123 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-7030-5479","authenticated-orcid":false,"given":"Beatrice","family":"Conte","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Environmental Virology, Department of Biology, University of Pisa, Via S. Zeno 35\/39, 56123 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0131-053X","authenticated-orcid":false,"given":"Annalaura","family":"Carducci","sequence":"additional","affiliation":[{"name":"Laboratory of Hygiene and Environmental Virology, Department of Biology, University of Pisa, Via S. Zeno 35\/39, 56123 Pisa, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.coesh.2020.09.003","article-title":"Surveillance of SARS-CoV-2 RNA in wastewater: Methods optimisation and quality control are crucial for generating reliable public health information","volume":"17","author":"Ahmed","year":"2020","journal-title":"Curr. Opin. Environ. Sci. Health"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"504","DOI":"10.1039\/D0EW00946F","article-title":"Reproducibility and sensitivity of 36 methods to quantify the SARS-CoV-2 genetic signal in raw wastewater: Findings from an interlaboratory methods evaluation in the U.S","volume":"7","author":"Pecson","year":"2021","journal-title":"Environ. Sci. Water Res. 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