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. 2024 Feb 22;14(5):682.
doi: 10.3390/ani14050682.

In Vitro Antiviral and Virucidal Activity of Ozone against Feline Calicivirus

Cristiana Catella  1 , Francesco Pellegrini  1 , Alice Carbonari  1 , Matteo Burgio  1 , Giovanni Patruno  1 , Annalisa Rizzo  1 , Claudia Maria Trombetta  2 , Jolanda Palmisani  3 , Vito Martella  1 , Michele Camero  1 , Gianvito Lanave  1
Affiliations

In Vitro Antiviral and Virucidal Activity of Ozone against Feline Calicivirus

Cristiana Catella et al. Animals (Basel). .

Abstract

The Caliciviridae family includes several viral pathogens of humans and animals, including norovirus (NoV), genus Norovirus, and feline calicivirus (FCV), genus Vesivirus. Due to their resistance in the environment, NoV and FCV may give rise to nosocomial infections, and indirect transmission plays a major role in their diffusion in susceptible populations. A pillar of the control of viruses resistant to an environment is the adoption of prophylaR1.6ctic measures, including disinfection. Since NoVs are not cultivatable in common cell cultures, FCV has been largely used as a surrogate of NoV for the assessment of effective disinfectants. Ozone (O3), a molecule with strong oxidizing properties, has shown strong microbicidal activity on bacteria, fungi, protozoa, and viruses. In this study, the virucidal and antiviral activities of an O3/O2 gas mixture containing O3 were tested at different concentrations (20, 35, and 50 μg/mL) for distinct contact times against FCV. The O3/O2 gas mixture showed virucidal and antiviral activities against FCV in a dose- and contact time-dependent fashion. Ozonation could be considered as a valid strategy for the disinfection of environments at risk of contamination by FCV and NoV.

Keywords: feline calicivirus; in vitro; norovirus; ozone; surrogate; virucidal activity.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Cytotoxic effect of O3 at 50 μg/mL on 24 h monolayer of Crandell Reese Feline Kidney (CRFK) cells with live-cell imaging (magnification 10×) at 30–90 s (A), 180 s (B), and 300 s (C). Exposure of CRFK cells to O3 at lower concentrations produced similar results.
Figure 2
Figure 2
Cytotoxicity of CRFK cells (express as percentage) treated with O3/O2 gas mixture containing O3 at 20 μg/mL (A), 35 μg/mL (B), and 50 μg/mL (C) plotted against contact times. The horizontal dotted line indicates the cytotoxicity threshold of (20% of cell death).
Figure 3
Figure 3
Antiviral activity of ozone (O3) at different concentrations (20, 35 and 50 μg/mL) against feline calicivirus (FCV). Treatment of infected cell monolayers with O3 (protocol A). Treatment of cell monolayers with O3 before virus infection (protocol B). FCV not treated (control virus, CV) and treated with O3 at room temperature for 30 s (A), 60 s, (B) and 90 s (C) were subsequently titrated on Crandell Reese Feline Kidney (CRFK) cells. Viral titers of FCV are expressed as log10 TCID50/50 μL. Significant p values are displayed. Bars in the figures indicate the means. Error bars indicate the standard deviation.
Figure 4
Figure 4
Virucidal activity of ozone (O3) at different concentrations (20, 35, and 50 μg/mL) against feline calicivirus (FCV). FCV was incubated with O3 for 30, 60, 90, 180, 300, and 600 s at room temperature. FCV not treated (control virus, CV) and treated with O3 were subsequently titrated on Crandell Rees Feline Kidney (CRFK) cells. Viral titers of FCV are expressed as log10 TCID50/50 μL. Significant p values are displayed. Bars in the figures indicate the means. Error bars indicate the standard deviation.
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