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Accordingly, to establish effective disinfection methods against AdVs, this study assessed and compared the virucidal efficacy of multiple virucidal agents, namely ozonated water (OW), povidone-iodine (PVPI), and ethanol (EtOH), against AdV type 40 in vitro. The AdV-inactivating activity of OW was influenced by ozone concentration and was adversely affected by the presence of proteins. PVPI demonstrated consistent virucidal activity across a concentration range of 0.025\u20130.050% and maintained its efficacy in the presence of proteins. EtOH showed time-dependent and limited concentration-dependent virucidal activity across a concentration range of 60\u201380% and maintained its effectiveness in the presence of proteins. Polymerase chain reaction analysis of the viral genome revealed that OW was the only agent among the three that induced AdV genome degradation. Therefore, the study findings indicate that if the properties of OW are comprehensively understood to ensure its appropriate use, OW can be applied as a useful virucidal agent against AdV as well as other virus-inactivating agents, such as PVPI and EtOH.<\/jats:p>","DOI":"10.2166\/wh.2025.009","type":"journal-article","created":{"date-parts":[[2025,7,7]],"date-time":"2025-07-07T11:47:10Z","timestamp":1751888830000},"page":"894-904","update-policy":"https:\/\/doi.org\/10.2166\/iwapcrossmarkpolicypage","source":"Crossref","is-referenced-by-count":1,"title":["Comparative evaluation of the virucidal activities of ozonated water, povidone-iodine, and ethanol against human adenovirus"],"prefix":"10.2166","volume":"23","author":[{"given":"Yohei","family":"Takeda","sequence":"first","affiliation":[{"name":"a Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan"},{"name":"b Department of Veterinary Medicine, Obihiro University of 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