Molecular surveillance confirms absence of avian influenza virus type A in Ecuadorian poultry: A retrospective reverse transcription quantitative polymerase chain reaction study from April 2023 to June 2024

Background and Aim: Avian influenza (Al) is a highly contagious zoonotic disease affecting birds and, occasionally, humans. Ecuador confirmed its first case of Al in late 2022, resulting in significant avian mortality and economic losses. In response, the Ecuadorian government implemented a mandatory control program emphasizing pre-vaccination diagnostics using reverse transcription quantitative polymerase chain reaction (RT-qPCR) or enzyme-linked immunosorbent assay. This study aimed to confirm the absence of Al virus type A in poultry across six major provinces of Ecuador following the 2022 outbreak, as part of the national eradication and vaccination campaign. Materials and Methods: A cross-sectional, retrospective molecular surveillance study was conducted from April 2023 to June 2024. A total of 343 pooled tracheal swab samples were collected from poultry farms in six provinces (Chimborazo, Cotopaxi, El Oro, Pastaza, Pichincha, and Tungurahua), covering over 1 million birds. The samples were analyzed using realtime RT-qPCR, targeting the M gene, and subtype-specific genes (H5, H7, and H7N9). Results: All 343 pooled samples (100%) tested negative for Al virus type A, suggesting an absence of active viral circulation during the study period. The implementation of pre-vaccination testing and biosecurity protocols contributed to this outcome. Conclusion: The study confirms that Al virus type A was not circulating in Ecuadorian poultry during the surveillance period. These findings underscore the effectiveness of collaborative efforts among government, industry, and laboratories. Ongoing molecular surveillance remains essential for early detection and prevention of future outbreaks, reinforcing Ecuador's commitment to One Health principles.