Characterization of clade 2.3.4.4 highly pathogenic H5 avian influenza viruses in ducks and chickens

doi:10.1016/j.vetmic.2015.11.001

Veterinary Microbiology

Volume 182, 15 January 2016, Pages 116-122

https://doi.org/10.1016/j.vetmic.2015.11.001 Get rights and content

Highlights

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Ducks are susceptible host of clade 2.3.4.4 H5 reassortant and the horizontal transmission among ducks were efficient.
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Clade 2.3.4.4 H5 reassortants were highly pathogenic in chickens.
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The current vaccines that are widely used in China may fail to confer protection against the H5 reassortants.

Abstract

Worldwide dissemination of reassortant variants of H5 clade 2.3.4.4 highly pathogenic avian influenza (HPAI) viruses has posed a great threat to the poultry industry. Here, we systematically characterized the H5N2, H5N6 and H5N8 influenza viruses in poultry and compared them with those of previous clade 2.3.4 H5N1 virus. All the three H5 subtype reassortants caused systematic infection in ducks, and exhibited efficient direct transmission in ducks. All of them were highly pathogenic in chickens; however, the H5 reassortants have reduced virulence compared to the parental H5N1 virus. Antigenicity analysis revealed that the current vaccines that are widely used in China may fail to confer protection against the H5 reassortants.

Introduction

HPAI H5N1 viruses continue circulating among avian species and have caused sporadically infection in humans, posing public concerns. The viruses have evolved into ten distinct phylogenetic clades (clades 0–9) on the basis of the HA gene since its occurrence in China in 1996 (Duan et al., 2008, Group, 2008). Among these clades, 2.3.4 is one of the main clade circulating in poultry (Jiang et al., 2010, Li et al., 2010) and has caused the most of confirmed human cases in China (Yu et al., 2008). Since 2010, H5HPAI viruses of clade 2.3.4 with different NA subtypes including H5N2 (Zhao et al., 2012b), H5N5 (Gu et al., 2011, Liu et al., 2013), H5N6 (Bi et al., 2015, Wu et al., 2015) and H5N8 (Fan et al., 2014a, Jeong et al., 2014, Kang et al., 2015, Lee et al., 2014, Wu et al., 2014b) were identified in domestic and wild waterfowl. The spread of HPAI H5 viruses in Asia, Europe and North America, which has been linked to the transcontinental migration of wild birds, has become a major zoonosis concern in the world (Verhagen et al., 2015b). As the HA gene of these viruses showing >1.5% average distances from other subclades of clade 2.3.4, the H5 Evolution Working Group defined it as a novel clade 2.3.4.4 (http://www.who.int/influenza/gisrs_laboratory/h5_nomenclature_clade2344/en/). So far, the pathogenesis of the novel clade viruses remains largely unknown. Therefore, we have characterized the antigenicity, pathogenicity and transmission of H5N2, H5N6 and H5N8 HPAI viruses in ducks and chickens.

Section snippets

Ethics statement

All animal research was approved by the Beijing Association for Science and Technology (approval ID SYXK, Beijing, 2007-0023) and performed in compliance with the Beijing Laboratory Animal Welfare and Ethics guidelines, as issued by the Beijing Administration Committee of Laboratory Animals, and in accordance with the China Agricultural University (CAU) Institutional Animal Care and Use Committee guidelines (ID:SKLAB-B-2010-003) approved by the Animal Welfare Committee of CAU. All experiments

Pathogenicity and transmissibility in ducks

Previous study indicated that clade 2.3.4 H5N1 viruses could replicate to high titers in the lungs and brains of ducks and killed all of the inoculated ducks (Li et al., 2010). In this study, all the six H5 reassortants replicated well in the lung, spleen, kidney and brain, causing systemic infection in ducks, which were similar to the H5N1 virus CN0603 (Fig. 1). Among these organs, lung and brain were the preferential tissues for viral growth. The virus titers in the brains of the H5N6 and

Discussion

Since 2010, H5HPAI viruses of different NA subtypes (H5N1, H5N2, H5N6, and H5N8) were increasingly detected in wild and domestic birds in Asian countries (Fan et al., 2014b, Gu et al., 2013, Wu et al., 2014a, Zhao et al., 2012a, Zhao et al., 2013a). The current situation of H5 virus emerging in Europe and North America since 2014 (Verhagen et al., 2015a) reveals that H5 virus is becoming a real global threat. Although there were some reports on these novel viruses previously, the comprehensive

Acknowledgements

This work was supported by Key Technologies Research and Development Program of China Grant (2013BAD12B01), National Science Fund for Distinguished Young Scholars (31025029), and Chinese Universities Scientific Fund (2015DY004)

References (27)

L. Duan et al.
The development and genetic diversity of H5N1 influenza virus in China, 1996–2006
Virology
(2008)
J. Jeong et al.
Highly pathogenic avian influenza virus (H5N8) in domestic poultry and its relationship with migratory birds in South Korea during 2014
Vet. Microbiol.
(2014)
C.G. Liu et al.
Emerging multiple reassortant H5N5 avian influenza viruses in ducks, China, 2008
Vet. Microbiol.
(2013)
H. Wu et al.
Novel reassortant highly pathogenic H5N6 avian influenza viruses in poultry in China
Infect. Genet. Evol.
(2015)
K. Zhao et al.
Characterization of three H5N5 and one H5N8 highly pathogenic avian influenza viruses in China
Vet. Microbiol.
(2013)
K. Zhao et al.
Characterization of three H5N5 and one H5N8 highly pathogenic avian influenza viruses in China
Vet. Microbiol.
(2013)
Y. Bi et al.
Two novel reassortants of avian influenza A (H5N6) virus in China
J. Gen. Virol.
(2015)
H. Chen
H5N1 avian influenza in China
Sci. China C Life Sci.
(2009)
H. Chen et al.
Development and application of avian influenza vaccines in China
Curr. Top. Microbiol. Immunol.
(2009)
S. Fan et al.
A novel highly pathogenic H5N8 avian influenza virus isolated from a wild duck in China
Influenza Other Respir. Viruses
(2014)

S. Fan et al.

A novel highly pathogenic H5N8 avian influenza virus isolated from a wild duck in China

Influenza Other Respir. Viruses

(2014)

W.O.F.H.N.E.W. Group

Toward a unified nomenclature system for highly pathogenic avian influenza virus (H5N1)

Emerg. Infect. Dis.

(2008)

M. Gu et al.

Novel reassortant highly pathogenic avian influenza (H5N5) viruses in domestic ducks, China

Emerg. Infect. Dis.

(2011)

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Genetics and pathogenicity of H5N6 highly pathogenic avian influenza viruses isolated from wild birds and a chicken in Japan during winter 2017–2018
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Therefore, further analysis of genome sequences of AIVs and migration of wild birds in Siberia is needed to elucidate how AIVs/HPAIVs move cross-continentally. The virulence of clade 2.3.4.4 H5 reassortants reportedly is lower than that of ancestral H5N1 HPAIVs (Lee al., 2016, 2017; Sun et al., 2016). In addition, our previous experimental infections of several H5 HPAIVs revealed that the lethal infectious dose of the Kagawa, Tokyo, and Hyogo strains in chickens is relatively low because 50% chicken lethal doses of them (104.38 to 104.83) were 10-fold more or higher than those of H5N1 HPAIVs during Winter 2010–2011 (Uchida et al., 2012), some H5N8 HPAIVs of Winter 2014–2015 (Kanehira et al., 2015; Tanikawa et al., 2016), and the H5N6 HPAIVs of Winter 2016–2017 (Takemae et al., 2017) under same experimental condition.
An H5N6 highly pathogenic avian influenza virus (HPAIV) outbreak occurred in poultry in Japan during January 2018, and H5N6 HPAIVs killed several wild birds in 3 prefectures during Winter 2017–2018. Time-measured phylogenetic analyses demonstrated that the Hemagglutinin (HA) and internal genes of these isolates were genetically similar to clade 2.3.4.4.B H5N8 HPAIVs in Europe during Winter 2016–2017, and Neuraminidase (NA) genes of the poultry and wild bird isolates were gained through distinct reassortments with AIVs that were estimated to have circulated possibly in Siberia during Summer 2017 and Summer 2016, respectively. Lethal infectious dose to chickens was similar between the poultry and wild-bird isolates. H5N6 HPAIVs during Winter 2017–2018 in Japan had higher 50% chicken lethal doses and lower transmission efficiency than the H5Nx HPAIVs that caused previous outbreaks in Japan, thus explaining in part why cases during the 2017–2018 outbreak were sporadic.
Full genome characterization of Iranian H5N8 highly pathogenic avian influenza virus from Hooded Crow (Corvus cornix), 2017: The first report
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Avian influenza viruses (AIVs) are not only important pathogens for birds but also they are important in public health due to cause sporadic and non-sustained infections in human populations. The H5 viruses have evolved into ten distinct phylogenetic clades (clades 0–9) by the HA gene of isolates from 1996 to 2006 in China [1]. H5N1 highly pathogenic avian influenza viruses (HPAIVs) of the A/Goose/Guangdong/1/1996 (Gs/Gd) lineage have become panzootic in domestic birds in Eurasia and Africa, since they detect for the first time) [2].
During 2014–2017 Clade 2.3.4.4 H5N8 highly pathogenic avian influenza viruses (HPAIVs) have spread worldwide. In 2016, an epidemic of HPAIV H5N8 in Iran caused mass deaths among wild birds, and several commercial poultry farms and captive bird holdings were affected and continue to experience problems. Several outbreaks were reported in 2017. One of them is related to Hooded crow (Corvus cornix) in a national park in Esfahan province in 2017. Whole genome sequencing and characterization have been done on the detected H5N8 sample. Based on HA sequencing results, it belongs to 2.3.4.4 clade, and the cleavage site is (PLREKRRKR/G). Phylogenetic analysis of the HA gene showed that the Iran 2017 H5N8 virus clustered within subgroup Russia 2016 2.3.4.4 b of group B in H5 clade 2.3.4.4 HPAIV.
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Previous studies indicated that the distribution of domestic duck density and wild waterfowl migration were important factors in HPAIV outbreaks (Gilbert et al., 2010; Hill et al., 2015). Further, many studies indicated that duck species could be infected by HPAIVs, especially clade 2.3.4.4 viruses, and shed a high titer of viruses without clinical signs (Hiono et al., 2017; Lee et al., 2016b; Pantin-Jackwood et al., 2017; Sun et al., 2016). Therefore, duck species may be important as a healthy reservoir in HPAIV epidemiology.
H5Nx clade 2.3.4.4 highly pathogenic avian influenza viruses (HPAIVs) have been disseminated to wide geographic regions since 2014. In 2016, five distinct genotypes (C-1 to C-5) of clade 2.3.4.4c H5N6 HPAIVs were detected in South Korea. In this study, we evaluated the pathogenicity, susceptibility to infection, and transmissibility of the two strains representing the C-1 and C-4 genotypes of the H5N6 viruses, which have different PA and NS gene, in domestic ducks. Although the susceptibility to infection of domestic ducks to the two strains was similar, the C-4 genotype virus induced higher mortality in ducks than C-1 genotype virus. A higher titer of viral shedding were detected in ducks challenged with the C-4 genotype virus compared with the C-1 genotype virus. These results indicated that the reassortment of HPAIVs with prevailing low pathogenic avian influenza viruses could effect on the pathogenicity in ducks.

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¹: These authors contributed equally to this work.

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Short communicationCharacterization of clade 2.3.4.4 highly pathogenic H5 avian influenza viruses in ducks and chickens

Highlights

Abstract

Introduction

Section snippets

Ethics statement

Pathogenicity and transmissibility in ducks

Discussion

Acknowledgements

Virology

Vet. Microbiol.

Vet. Microbiol.

Infect. Genet. Evol.

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Two novel reassortants of avian influenza A (H5N6) virus in China

J. Gen. Virol.

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Sci. China C Life Sci.

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Curr. Top. Microbiol. Immunol.

A novel highly pathogenic H5N8 avian influenza virus isolated from a wild duck in China

Influenza Other Respir. Viruses

A novel highly pathogenic H5N8 avian influenza virus isolated from a wild duck in China

Influenza Other Respir. Viruses

Toward a unified nomenclature system for highly pathogenic avian influenza virus (H5N1)

Emerg. Infect. Dis.

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Emerg. Infect. Dis.

Short communication
Characterization of clade 2.3.4.4 highly pathogenic H5 avian influenza viruses in ducks and chickens