Antiviral Activity of Essential Oils Against Avian Influenza Virus H7N3 In Vitro and In Ovo Models

The poultry industry is continually seeking efficient, practical strategies to control infectious diseases. Among these new alternatives are essential oils (EOs), naturally occurring compounds with antimicrobial properties. Their effectiveness has been demonstrated in various studies that focus on their broad antiviral properties. The present experiment evaluated the antiviral efficacy of an EOs formulation against the H7N3 subtype of avian influenza virus (AIV) by directly mixing virus and EOs (virus/EOs mixture) through an in vitro model in cultured Madin-Darby canine kidney cells (MDCKs). The experiment used a focus reduction neutralization test (FRNT) to determine the 50% inhibitory concentration (IC₅₀) by virus/EOs mixture, as well as the application of EOs 24 h before infection, through a viral inhibition test using a chicken embryo (CE) in ovo model. The results demonstrated the antiviral activity of the EOs formulation against the H7N3 in vitro model (IC₅₀ values of 20.4 and 38.3 ppm and selective index (SI) values of 9.4 and 5.1) and in ovo model (decreasing hemagglutination titers to 1 HA unit, 10^5.28 embryo infectious dose 50% (EID₅₀) per mL, and viral loads to approximately 10^11.4 copies/mL) when applied in CE, 24 h before viral infection, representing the lowest replication indicators recorded during the experiment. According to the results, the EOs formulation demonstrated antiviral activity against AIV H7N3 both as a virus/EOs mixture and through application in ovo 24 h before infection. Application 24 h before infection in CE showed a significant effect compared with the virus/EOs mixture, demonstrating an antiviral effect in the ovo infection model. This study demonstrates both the virucidal and antiviral capacity of the compounds in the EOs formulation against AIV H7N3 and their efficacy when applied 24 h before infection in the in ovo model.