Continuing evolution of H9N2 influenza viruses in Southeastern China

Abstract

H9N2 influenza viruses are panzootic in domestic poultry in Eurasia and since 1999 have caused transient infections in humans and pigs. To investigate the zoonotic potential of H9N2 viruses, we studied the evolution of the viruses in live-poultry markets in Hong Kong in 2003. H9N2 was the most prevalent influenza virus subtype in the live-poultry markets between 2001 and 2003. Antigenic and phylogenetic analysis of hemagglutinin (HA) showed that all of the 19 isolates found except one belonged to the lineage represented by A/Duck/Hong Kong/Y280/97 (H9N2). The exception was A/Guinea fowl/NT184/03 (H9N2), whose HA is most closely related to that of the human isolate A/Guangzhou/333/99 (H9N2), a virus belonging to the A/Chicken/Beijing/1/94-like (H9N2) lineage. At least six different genotypes were recognized. The majority of the viruses had nonstructural (and HA) genes derived from the A/Duck/Hong Kong/Y280/97-like virus lineage but had other genes of mixed avian virus origin, including genes similar to those of H5N1 viruses isolated in 2001. Viruses of all six genotypes of H9N2 found were able to replicate in chickens and mice without adaptation. The infected chickens showed no signs of disease, but representatives of two viral genotypes were lethal to mice. Three genotypes of virus replicated in the respiratory tracts of swine, which shed virus for at least 5 days. These results show an increasing genetic and biologic diversity of H9N2 viruses in Hong Kong and support their potential role as pandemic influenza agents.

FIG. 1.

To determine the genetic diversity of the 19 virus isolates, we compared partial sequences of their eight gene segments. We identified six different genotypes, designated A, B, C, D, E, and F. Ph, pheasant.

FIG. 2.

(a) Phylogenetic analysis of the HA1 region of the H9 gene of the 19 H9N2 viruses isolated in 2003. As predicted by antigenic comparison, all but GF/HK/NT184/03 belonged to the Dk/HK/Y280/97-like lineage. (b) The N2 genes of the 19 isolates clustered into two main sublineages. Nine viruses clustered with Dk/HK/Y280/97-like viruses (which contain a three-amino-acid deletion in the stalk region), and 10 viruses grouped with Ck/HK/G9/97-like viruses (which do not contain a stalk deletion). Isolates in bold type represent isolates sequenced in this study. Ph, pheasant.