Isolation and characterization of genotype 4 Eurasian avian-like H1N1 influenza virus in pigs suffering from pneumonia

Swine influenza viruses pose ongoing threat to pork industry throughout the world. In 2023, fattening pigs from a swine farm in Inner Mongolia of China experienced influenza-like symptoms. Co-infection of influenza A virus with Pasteurella multocida was diagnosed in lung tissues of diseased pigs and a genotype 4 (G4) Eurasian avian-like (EA) H1N1 virus was isolated, which was named as A/swine/Neimenggu/0326/2023. We demonstrated the virus preferentially bound human-like SAα2,6Gal receptor. It was noteworthy that the virus possessed multiple genetic markers for mammalian adaptation in the internal genes. Animal studies showed that compared with genotype 1 (G1) EA H1N1 virus and early prevalent G4 EA H1N1 virus, A/swine/Neimenggu/0326/2023 virus exhibited increased virus shedding, enhanced replication in lungs, and caused more severe lung lesions in pigs. These findings indicate that the G4 EA H1N1 virus poses increased threat to pork industry, controlling the prevailing viruses in pigs should be promptly implemented. •A swine influenza virus (SW/NMG/0326/23) was isolated from dead pig suffering from severe pneumonia.•SW/NMG/0326/23 virus belonged to G4 genotype Eurasian avian-like (EA) H1N1 influenza virus.•SW/NMG/0326/23 virus had multiple amino acid substitutions that might enhance virus pathogenicity to mammals.•SW/NMG/0326/23 virus was more pathogenic and could cause more severe lung lesions in pigs.