The highly pathogenic avian influenza (HPAI) virus A/goose/Guangdong/1/96 H5N1 (Gs/GD) lineage has been transmitted globally and has caused deaths in wild birds, poultry, and humans. Clade, one of the subclades of the Gs/GD lineage, spread through Taiwan in late 2014 and become an endemic virus. We analyzed 239 newly sequenced HPAI clade H5Nx isolates to explore the phylogenetic relationships, divergence times, and evolutionary history of Taiwan HPAI H5Nx viruses from 2015 to 2018. Overall, 15 reassortant genotypes were identified among H5N2, H5N3, and H5N8 viruses. Maximum likelihood and Bayesian phylogenies based on homologous hemagglutinin (HA) and matrix protein (MP) genes suggest that Taiwan HPAI H5Nx viruses share a most recent common ancestor that has diversified since October 2014 and is closely related to two HPAI H5N8 viruses identified from wild birds in Japan. Two waves of HPAI caused by multiple reassortants were identified, the first occurring in late 2014 and the second beginning in late 2016. The first wave consisted of seven H5Nx reassortants that spread through Taiwan. In the second wave, eight novel reassortants were detected which had newly introduced internal genes, mostly derived from the avian influenza virus gene pool maintained in wild birds in Asia. Phylodynamic reconstruction using the Bayesian Skygrid model revealed varied fluctuating patterns among reassortants. The mean evolutionary rate also varied among reassortants and subtypes. The neuraminidase (NA) gene evolved faster than the HA gene in H5N2 viruses, while HA evolved faster than NA in H5N8 viruses. The HA mean evolutionary rate ranged from 6.00 × 10 to 7.73 × 10 and from 5.81 × 10 to 9.45 × 10 substitutions/site/year for H5N2 and H5N8 viruses, respectively. The continuous circulation of HPAI H5Nx variants and the emergence of novel reassortants in Taiwan highlight that the surveillance, biosecurity, and management systems of poultry farms need to be improved and carefully executed.
Copyright © 2021. Published by Elsevier B.V.