Background: Since the early 2000s, Avian influenza virus subtype H9N2 has been widely circulating in North African countries, including Libya [1], Tunisia [2], and Egypt [3]. For uncertain reasons, H9N2 was not detected in Moroccan farms until the end of 2016 [4] and at present it is endemic in poultry. This study was carried out to evaluate the evolution of H9 AI viruses in Morocco from 2017 to 2019. Methods: In this study, 16 H9 viruses collected from 2017 to 2019 in Morocco were isolated and sequenced. The genomic signatures and protein sequences of these isolates were analyzed. Results: Moroccan H9 viruses were closely related to viruses isolated from African and Middle Eastern countries, and their HA gene resembled those of viruses of the G1-like lineage, multiple mammalian-host-associated mutations were detected that favor transmission from avian to mammalian hosts. Other mutations related to virulence were also identified. Conclusion : Our findings suggest that an attention should be paid to the control of H9N2 influenza viruses in chickens and a continuous monitoring of the prevalence and pathogenic potential of AIV H9N2 in the poultry population in Morocco. Keywords: Influenza Virus, H9, Phylogenetic analyses, Morocco

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