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Dual Antiviral and Virucidal Effects of Chlorine Dioxide on Influenza A Virus

Shin, Bo-Hye; Song, Kyoung Ju; Kim, Jong Rak; Kim, Kyongmin
Department of Microbiology, Ajou University School of Medicine
Chlorine dioxide, ClO2, gas is a powerful disinfectant which is 2.5 and 500,000 times more effective than chlorine-based disinfectants and alcohol, respectively. In this study, we examined whether chlorine dioxide inhibits the proliferations of Influenza A viruses and then how effective this chlorine dioxide to kill the influenza A viruses. Chlorine dioxide gas was induced from “farm-e Tok”. “Farm-e Tok” is a ready-to-use product, without necessitating ClO2 gas generator on site. The treated concentration of ClO2 was 0.16ppmv/min. First, we inoculated influenza A viruses into 11-day old embryonated eggs and incubated one more day at 37°C. Next day, we exposed chlorine dioxide to virus-inoculated experimental eggs but not to virus-inoculated control eggs, and incubated further for 48 hrs at 37°C. At 3 days post-infection, amniotic fluids were collected to harvest Influenza A viruses and titrated by plaque assay on Madin-Darby canine kidney (MDCK) cells. Titer of control viruses reached 6.8 x 105 plaque forming unit (PFU). Whereas, titer of experimental eggs, which were exposed with chlorine dioxide for 2 days, was 1.0 x 105 PFU, demonstrating that chlorine dioxide gas inhibits the proliferation of Influenza A viruses. On the second experiment, we exposed chlorine dioxide gas directly to the aliquots of influenza A viruses to examine the virucidal effect of chlorine dioxide. To start this experiment, we activated chlorine dioxide batch for 11 hrs, then aliquots of viruses were exposed to chlorine dioxide gas for 1, 6, or 12 hrs, respectively and titrated by plaque assay on MDCK cells. Virus titers were about 2,000 times lower after 6 hrs exposure and more than 3-logs lower after 12 hrs exposure, however control virus titers were slightly deceased with 4.6 times after 12 hrs. Taken together, we suggest that chlorine dioxide gas has dual antiviral and virucidal effects on influenza A viruses.
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