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. 2015 Jun 1:5:10130.
doi: 10.1038/srep10130.

The infection of turkeys and chickens by reassortants derived from pandemic H1N1 2009 and avian H9N2 influenza viruses

Honglei Sun  1 , Weili Kong  1 , Litao Liu  1 , Yi Qu  1 , Chong Li  1 , Ye Shen  1 , Yu Zhou  1 , Yu Wang  1 , Sizhe Wu  1 , Juan Pu  1 , Jinhua Liu  1 , Yipeng Sun  1
Affiliations

The infection of turkeys and chickens by reassortants derived from pandemic H1N1 2009 and avian H9N2 influenza viruses

Honglei Sun et al. Sci Rep. .

Abstract

Outbreaks of pandemic H1N1 2009 (pH1N1) in turkeys have been reported in several countries. Co-infection of pH1N1 and avian H9N2 influenza viruses in turkeys provide the opportunity for their reassortment, and novel reassortant viruses might further be transmitted to other avian species. However, virulence and transmission of those reassortant viruses in poultry remain unclear. In the present study, we generated 16 single-gene reassortant influenza viruses including eight reassortants on the pH1N1 background by individual replacement with a corresponding gene segment from H9N2 and eight reassortants on the H9N2 background replaced individually with corresponding gene from pH1N1, and characterized reassortants viruses in turkeys and chickens. We found that the pH1N1 virus dramatically increased its infectivity and transmissibility in turkeys and chickens after introducing any gene (except for PB2) from H9N2 virus, and H9N2 virus acquired single gene (except for HA) of pH1N1 almost did not influence its replication and transmission in turkeys and chickens. Additionally, 13 reassortant viruses transmitted from turkeys to chickens. Our results indicate that turkeys and chickens are susceptible to pH1N1-H9N2 reassortant viruses, and mixing breeding of different avian species would facilitate the transmission of these reassortant viruses.

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Figures

Figure 1
Figure 1. Viral growth kinetics of wild type and recombinant viruses in CEF cells.
CEF cells were infected with BJ09, HB08 and their “7 + 1” reassortant viruses at an MOI of 0.001. Error bars above represent standard deviations from the means for three independent experiments.
Figure 2
Figure 2. Polymerase activity of 8 RNP combinations between the BJ09 and HB08 viruses.
The DF1 cells were transfected in duplicate with luciferase reporter plasmid p-Luci and internal control plasmid Renilla, together with plasmids expressing PB2, PB1, PA, and NP from either BJ09 or HB08 viruses. Segments derived from pH1N1 are in yellow, and those derived from HB08 virus are in blue. Cells were incubated at 37 °C (stippled bars) or at 39 °C (solid bars) for 24 h, and cell lysates were analyzed to measure Firefly and Renilla luciferase activities. Values shown are the mean ± SD of the three independent experiments and are standardized to those of HB08 measured at 37 °C (100%). * and ** indicate that the polymerase activity between 37 °C and 39 °C was significantly different with P < 0.05 and P < 0.01, respectively (ANOVA). PB2, PB1, PA, and NP expressions were detected by Western blot analysis.
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