Phylogenetic analysis and pathogenicity of H3 subtype avian influenza viruses isolated from live poultry markets in China
Identifieur interne : 000104 ( Pmc/Curation ); précédent : 000103; suivant : 000105Phylogenetic analysis and pathogenicity of H3 subtype avian influenza viruses isolated from live poultry markets in China
Auteurs : Hongrui Cui [République populaire de Chine] ; Ying Shi [République populaire de Chine] ; Tao Ruan [République populaire de Chine] ; Xuesong Li [République populaire de Chine] ; Qiaoyang Teng [République populaire de Chine] ; Hongjun Chen [République populaire de Chine] ; Jianmei Yang [République populaire de Chine] ; Qinfang Liu [République populaire de Chine] ; Zejun Li [République populaire de Chine]Source :
- Scientific Reports [ 2045-2322 ] ; 2016.
Abstract
H3 subtype influenza A virus is one of the main subtypes that threats both public and animal health. However, the evolution and pathogenicity of H3 avian influenza virus (AIV) circulating in domestic birds in China remain largely unclear. In this study, seven H3 AIVs (four H3N2 and three H3N8) were isolated from poultry in live poultry market (LPM) in China. Phylogenetic analyses of full genomes showed that all viruses were clustered into Eurasian lineage, except N8 genes of two H3N8 isolates fell into North American lineage. Intriguingly, the N8 gene of one H3N8 and PB2, PB1, NP and NS of two H3N2 isolates have close relationship with those of the highly pathogenic H5N8 viruses circulating in Korea and United States, suggesting that the H3-like AIV may contribute internal genes to the highly pathogenic H5N8 viruses. Phylogenetic tree of HA gene and antigenic cross-reactivity results indicated that two antigenically different H3 viruses are circulating in LPM in China. Most of the H3 viruses replicated in mice lung and nasal turbinate without prior adaptation, and the representative H3 viruses infected chickens without causing clinical signs. The reassortment of H3 subtype influenza viruses warrants continuous surveillance in LPM in China.
Url:
DOI: 10.1038/srep27360
PubMed: 27270298
PubMed Central: 4895239
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<front><div type="abstract" xml:lang="en"><p>H3 subtype influenza A virus is one of the main subtypes that threats both public and animal health. However, the evolution and pathogenicity of H3 avian influenza virus (AIV) circulating in domestic birds in China remain largely unclear. In this study, seven H3 AIVs (four H3N2 and three H3N8) were isolated from poultry in live poultry market (LPM) in China. Phylogenetic analyses of full genomes showed that all viruses were clustered into Eurasian lineage, except N8 genes of two H3N8 isolates fell into North American lineage. Intriguingly, the N8 gene of one H3N8 and PB2, PB1, NP and NS of two H3N2 isolates have close relationship with those of the highly pathogenic H5N8 viruses circulating in Korea and United States, suggesting that the H3-like AIV may contribute internal genes to the highly pathogenic H5N8 viruses. Phylogenetic tree of HA gene and antigenic cross-reactivity results indicated that two antigenically different H3 viruses are circulating in LPM in China. Most of the H3 viruses replicated in mice lung and nasal turbinate without prior adaptation, and the representative H3 viruses infected chickens without causing clinical signs. The reassortment of H3 subtype influenza viruses warrants continuous surveillance in LPM in China.</p>
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</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id>
<journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id>
<journal-title-group><journal-title>Scientific Reports</journal-title>
</journal-title-group>
<issn pub-type="epub">2045-2322</issn>
<publisher><publisher-name>Nature Publishing Group</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">27270298</article-id>
<article-id pub-id-type="pmc">4895239</article-id>
<article-id pub-id-type="pii">srep27360</article-id>
<article-id pub-id-type="doi">10.1038/srep27360</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Phylogenetic analysis and pathogenicity of H3 subtype avian influenza viruses isolated from live poultry markets in China</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Cui</surname>
<given-names>Hongrui</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Shi</surname>
<given-names>Ying</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Ruan</surname>
<given-names>Tao</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Xuesong</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Teng</surname>
<given-names>Qiaoyang</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chen</surname>
<given-names>Hongjun</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yang</surname>
<given-names>Jianmei</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Liu</surname>
<given-names>Qinfang</given-names>
</name>
<xref ref-type="corresp" rid="c1">a</xref>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Li</surname>
<given-names>Zejun</given-names>
</name>
<xref ref-type="corresp" rid="c2">b</xref>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<aff id="a1"><label>1</label>
<institution>Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences</institution>
, Shanghai,<country>People’s Republic of China</country>
</aff>
</contrib-group>
<author-notes><corresp id="c1"><label>a</label>
<email>liuqinfang@shvri.ac.cn</email>
</corresp>
<corresp id="c2"><label>b</label>
<email>lizejun@shvri.ac.cn</email>
</corresp>
</author-notes>
<pub-date pub-type="epub"><day>07</day>
<month>06</month>
<year>2016</year>
</pub-date>
<pub-date pub-type="collection"><year>2016</year>
</pub-date>
<volume>6</volume>
<elocation-id>27360</elocation-id>
<history><date date-type="received"><day>02</day>
<month>02</month>
<year>2016</year>
</date>
<date date-type="accepted"><day>12</day>
<month>05</month>
<year>2016</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2016, Macmillan Publishers Limited</copyright-statement>
<copyright-year>2016</copyright-year>
<copyright-holder>Macmillan Publishers Limited</copyright-holder>
<license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>
</license-p>
</license>
</permissions>
<abstract><p>H3 subtype influenza A virus is one of the main subtypes that threats both public and animal health. However, the evolution and pathogenicity of H3 avian influenza virus (AIV) circulating in domestic birds in China remain largely unclear. In this study, seven H3 AIVs (four H3N2 and three H3N8) were isolated from poultry in live poultry market (LPM) in China. Phylogenetic analyses of full genomes showed that all viruses were clustered into Eurasian lineage, except N8 genes of two H3N8 isolates fell into North American lineage. Intriguingly, the N8 gene of one H3N8 and PB2, PB1, NP and NS of two H3N2 isolates have close relationship with those of the highly pathogenic H5N8 viruses circulating in Korea and United States, suggesting that the H3-like AIV may contribute internal genes to the highly pathogenic H5N8 viruses. Phylogenetic tree of HA gene and antigenic cross-reactivity results indicated that two antigenically different H3 viruses are circulating in LPM in China. Most of the H3 viruses replicated in mice lung and nasal turbinate without prior adaptation, and the representative H3 viruses infected chickens without causing clinical signs. The reassortment of H3 subtype influenza viruses warrants continuous surveillance in LPM in China.</p>
</abstract>
</article-meta>
</front>
<floats-group><fig id="f1"><label>Figure 1</label>
<caption><title>Growth properties of the H3 viruses in MDCK, A549 and DF-1 cells.</title>
<p>The cells were infected with each virus at an MOI of 0.01. Each point on the curve indicates the mean from three independent experiments, and error bars indicates the standard errors of the mean (SEM) (*P < 0.05; **P < 0.01).</p>
</caption>
<graphic xlink:href="srep27360-f1"></graphic>
</fig>
<fig id="f2"><label>Figure 2</label>
<caption><title>Phylogenetic trees of the H3, N2, and N8 genes of the seven H3 influenza viruses.</title>
<p>The trees were generated by the distance based neighbor-joining method in the software MEGA 6.0. The reliability of the trees was assessed by bootstrap analysis with 1,000 replications. Horizontal distances are proportional to genetic distance. The viruses isolated in this study are in bold and red.</p>
</caption>
<graphic xlink:href="srep27360-f2"></graphic>
</fig>
<fig id="f3"><label>Figure 3</label>
<caption><title>Phylogenetic trees of the PB2, PB1, and PA genes of the seven H3 influenza viruses.</title>
</caption>
<graphic xlink:href="srep27360-f3"></graphic>
</fig>
<fig id="f4"><label>Figure 4</label>
<caption><title>Phylogenetic trees of the NP, M, and NS genes of the seven H3 influenza viruses.</title>
</caption>
<graphic xlink:href="srep27360-f4"></graphic>
</fig>
<fig id="f5"><label>Figure 5</label>
<caption><title>Replication of influenza virus in nasal turbinate and lung of inoculated mice.</title>
<p>Viral titers in the lungs (<bold>A,C</bold>
) and nasal turbinate (<bold>B,D</bold>
) harvested at 3, and 5dpi. Viral titers were determined by EID<sub>50</sub>
by using embryonated chicken eggs. Error bars represent standard errors of the mean (SEM), dotted line represents the limitation of the detection. *P < 0.05, **P < 0.01.</p>
</caption>
<graphic xlink:href="srep27360-f5"></graphic>
</fig>
<fig id="f6"><label>Figure 6</label>
<caption><title>Microscopic lung sections from mice inoculated with various H3 viruses on 5 dpi.</title>
<p>(<bold>A</bold>
) 74-1/H3N2, damage of alveolar epithelium (arrow), lymphocytic cuffing with marked lymphocyte infiltration of the lamina propria (asterisk). (<bold>B</bold>
) B1645-2/H3N2, mild bronchiolar epithelial damadge (arrow), mild alveolar septal thickening and light infiltration of lymphocyte (asterisk). (<bold>C</bold>
) B1646-2/H3N2, bronchiolar epithelial attenuation (arrow), peribronchiolar lymphocytic cuffing with marked lymphocyte infiltration (asterisk). (<bold>D</bold>
) B1647-1/H3N2, Moderate alveolar septal thickening and infiltration of lymphocyte (arrow). (<bold>E</bold>
) 120-1/H3N8, light alveolar septal thickening (arrow). (<bold>F</bold>
) A1591-1/H3N8, damage of alveolar epithelium (arrow), a large part of alveolar spaces were filled with lymphocyte (asterisk). (<bold>G</bold>
) B854-2/H3N8, severe damage of alveolar epithelium (arrow), alveolar lumina and septa were filled with a number of lymphocyte (asterisk). (<bold>H</bold>
) Mock. The scale bar indicates 50 μm in all photographs. H&E staining of the microscopic lung sections from mice necropsied at 5 dpi.</p>
</caption>
<graphic xlink:href="srep27360-f6"></graphic>
</fig>
<table-wrap position="float" id="t1"><label>Table 1</label>
<caption><title>Antigenic analysis of H3 avian influenza viruses.</title>
</caption>
<table frame="hsides" rules="groups" border="1"><colgroup><col align="left"></col>
<col align="center"></col>
<col align="center"></col>
<col align="center"></col>
<col align="center"></col>
</colgroup>
<thead valign="bottom"><tr><th rowspan="2" align="left" valign="bottom" charoff="50">Virus</th>
<th colspan="4" align="center" valign="top" charoff="50">HI titers to antiserum<hr></hr>
</th>
</tr>
<tr><th align="center" valign="top" charoff="50">Chicken sera against 74-1/H3N2<xref ref-type="fn" rid="t1-fn1">†</xref>
</th>
<th align="center" valign="top" charoff="50">Chicken sera against 120-1/H3N8</th>
<th align="center" valign="top" charoff="50">Chicken sera against 854-2/H3N</th>
<th align="center" valign="top" charoff="50">Chicken sera against B1646-2/H3N2</th>
</tr>
</thead>
<tbody valign="top"><tr><td align="left" valign="top" charoff="50">74-1/H3N2</td>
<td align="center" valign="top" charoff="50">256</td>
<td align="center" valign="top" charoff="50">1024</td>
<td align="center" valign="top" charoff="50">1024</td>
<td align="center" valign="top" charoff="50">64</td>
</tr>
<tr><td align="left" valign="top" charoff="50">120-1/H3N8</td>
<td align="center" valign="top" charoff="50">256</td>
<td align="center" valign="top" charoff="50">1024</td>
<td align="center" valign="top" charoff="50">1024</td>
<td align="center" valign="top" charoff="50">128</td>
</tr>
<tr><td align="left" valign="top" charoff="50">854-2/H3N8</td>
<td align="center" valign="top" charoff="50">128</td>
<td align="center" valign="top" charoff="50">1024</td>
<td align="center" valign="top" charoff="50">1024</td>
<td align="center" valign="top" charoff="50">64</td>
</tr>
<tr><td align="left" valign="top" charoff="50">B1646-2/H3N2</td>
<td align="center" valign="top" charoff="50">128</td>
<td align="center" valign="top" charoff="50">128</td>
<td align="center" valign="top" charoff="50">128</td>
<td align="center" valign="top" charoff="50">512</td>
</tr>
</tbody>
</table>
<table-wrap-foot><fn id="t1-fn1"><p><sup>†</sup>
Antisera were generated from SPF chickens immunized twice with representative H3 avian influenza viruses. Homologous titers are shown in boldface.</p>
</fn>
</table-wrap-foot>
</table-wrap>
</floats-group>
</pmc>
</record>
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