Pathobiological Characterization of a Novel Reassortant Highly Pathogenic H5N1 Virus Isolated in British Columbia, Canada, 2015.

Yohannes Berhane^1,2 Darwyn Kobasa^3,4 Carissa Embury-Hyatt¹ Brad Pickering¹ Shawn Babiuk^1,5 Tomy Joseph⁶ Victoria Bowes⁶ Mathew Suderman¹ Anders Leung³ Colleen Cottam-Birt¹ Tamiko Hisanaga¹ John Pasick¹

Affiliations 6 institutions

Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Winnipeg, Manitoba, Canada R3E 3M4.
Department of Animal Science, University of Manitoba, Winnipeg, Canada.
Public Health Agency of Canada, National Microbiology Laboratory, 1015 Arlington Street, Winnipeg, Manitoba, Canada.
Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada.
Department of Immunology, University of Manitoba, Winnipeg, Canada.
Animal Health Centre, Ministry of Agriculture, Abbotsford, British Columbia, Canada V3G 2M3.

PMID26988892 DOI10.1038/srep23380 Year2016 Journal Scientific reports Sci Rep Languageeng Statusepublish

Abstract

In the current study, we describe the pathobiologic characteristics of a novel reassortant virus - A/chicken/BC/FAV-002/2015 (H5N1) belonging to clade 2.3.4.4 that was isolated from backyard chickens in British Columbia, Canada. Sequence analyses demonstrate PB1, PA, NA and NS gene segments were of North American lineage while PB2, HA, NP and M were derived from a Eurasian lineage H5N8 virus. This novel virus had a 19 amino acid deletion in the neuraminidase stalk. We evaluated the pathogenic potential of this isolate in various animal models. The virus was highly pathogenic to mice with a LD50 of 10 plaque forming units (PFU), but had limited tissue tropism. It caused only subclinical infection in pigs which did result in seroconversion. This virus was highly pathogenic to chickens, turkeys, juvenile Muscovy ducks (Cairnia moschata foma domestica) and adult Chinese geese (Anser cynoides domesticus) causing a systemic infection in all species. The virus was also efficiently transmitted and resulted in mortality in naïve contact ducks, geese and chickens. Our findings indicate that this novel H5N1 virus has a wide host range and enhanced surveillance of migratory waterfowl may be necessary in order to determine its potential to establish itself in the wild bird reservoir.

Supporting text Virus Host Location

Amino Acid Sequence 128 Animals 1948 Animals, Wild 187 British Columbia 6 Chickens 146 Ducks 81 Evolution, Molecular 176 Geese 21 Influenza A Virus, H5N1 Subtype 300 Mice 253 Neuraminidase 62 Orthomyxoviridae Infections 228 Phylogeny 805 Reassortant Viruses 103 Sequence Deletion 8 Swine 258 Viral Load 36 Viral Tropism 45

Evidence records

8 total

Zoonotic Surveillance

1 records · 1 evidence types

Evidence type Virus Isolation

1 records

OVE2297 Virus Isolation

Key finding

A novel reassortant H5N1 virus, A/chicken/BC/FAV-002/2015, was successfully isolated from backyard chickens in British Columbia, Canada.

Virus

Host

Location

Supporting text

A novel reassortant virus - A/chicken/BC/FAV-002/2015 (H5N1) ... was isolated from backyard chickens in British Columbia, Canada.

Sample type: chicken specimens

Experimental Infection

6 records · 2 evidence types

Evidence type Pathogenicity Experiment

5 records

OVE2299 Pathogenicity Experiment

Key finding

A/chicken/BC/FAV-002/2015 (H5N1) was highly pathogenic to mice with an LD50 of 10 PFU, showing severe virulence but limited tissue tropism in the mouse infection model.

Virus

Host

Location

Supporting text

The virus was highly pathogenic to mice with a LD50 of 10 plaque forming units (PFU), but had limited tissue tropism.

Method: experimental infection | LD50 determination
Experimental system: mouse infection model

OVE2300 Pathogenicity Experiment

Key finding

A/chicken/BC/FAV-002/2015 (H5N1) was highly pathogenic to chickens causing systemic infection.

Virus

Host

Location

Supporting text

This virus was highly pathogenic to chickens, turkeys, juvenile Muscovy ducks (Cairnia moschata foma domestica) and adult Chinese geese (Anser cynoides domesticus) causing a systemic infection in all species.

Method: experimental infection | pathological examination
Experimental system: chicken infection experiment

OVE2301 Pathogenicity Experiment

Key finding

A/chicken/BC/FAV-002/2015 (H5N1) was highly pathogenic to turkeys causing systemic infection.

Virus

Host

Location

Supporting text

Method: experimental infection | pathological examination
Experimental system: turkey infection experiment

OVE2302 Pathogenicity Experiment

Key finding

A/chicken/BC/FAV-002/2015 (H5N1) was highly pathogenic to juvenile Muscovy ducks (Cairnia moschata foma domestica) causing systemic infection.

Virus

Host

Location

Supporting text

Method: experimental infection | pathological examination
Experimental system: duck infection experiment

OVE2303 Pathogenicity Experiment

Key finding

A/chicken/BC/FAV-002/2015 (H5N1) was highly pathogenic to adult Chinese geese (Anser cynoides domesticus) causing systemic infection.

Virus

Host

Location

Supporting text

Method: experimental infection | pathological examination
Experimental system: goose infection experiment

Evidence type Experimental Transmission

1 records

OVE2305 Experimental Transmission

Key finding

A/chicken/BC/FAV-002/2015 (H5N1) virus was efficiently transmitted from experimentally infected birds to naïve contact ducks, geese, and chickens, causing mortality in recipients.

Virus

Host

waterfowl Anatidae Experimental system

geese Anser Experimental system

chicken Gallus gallus Experimental system

Location

Supporting text

The virus was also efficiently transmitted and resulted in mortality in naïve contact ducks, geese and chickens.

Method: animal inoculation | co-housing contact exposure | observation of clinical signs | mortality assessment
Experimental system: contact transmission model in co-housed birds under controlled laboratory conditions

Genomic Evolution

1 records · 1 evidence types

Evidence type Recombination or Reassortment

1 records

OVE2298 Recombination or Reassortment

Key finding

The H5N1 virus A/chicken/BC/FAV-002/2015 was a reassortant containing North American lineage PB1, PA, NA, and NS segments and Eurasian H5N8–derived PB2, HA, NP, and M segments.

Virus

Host

Location

Supporting text

Sequence analyses demonstrate PB1, PA, NA and NS gene segments were of North American lineage while PB2, HA, NP and M were derived from a Eurasian lineage H5N8 virus.

Event type: reassortment
Genes or segments: PB1 | PA | NA | NS | PB2 | HA | NP | M

References

43 references

Reference network

Force-directed citation graph. OmniVira-indexed references are prioritized and recursively expanded up to three steps.

194 nodes · 225 links

Drag nodes to explore citation neighborhoods


PMID 25804829	Reassortant Highly Pathogenic Influenza A H5N2 Virus Containing Gene Segments Related to Eurasian H5N8 in British Columbia, Canada, 2014	Pasick	2015
PMID 21408010 In OmniVira	Wild bird migration across the Qinghai-Tibetan plateau: a transmission route for highly pathogenic H5N1.	Prosser	2011
-	Multicontinental epidemic of H5N1 HPAI virus (1996–2007). In Avian Influenza	Sims L. D. & Brown	2008
PMID 18394278	Wild ducks as long-distance vectors of highly pathogenic avian influenza virus (H5N1)	Keawcharoen	2008
PMID 10484749	Y., Subbarao K., Cox N	Xu	1999
PMID 26038499 In OmniVira	Pathobiological features of a novel, highly pathogenic avian influenza A(H5N8) virus.	Kim	2014
PMID 25776760	Pathogenesis of novel reassortant avian influenza virus A (H5N8) isolates in the ferret. Virology 481, 136–141 (2015)	Kim	2015
PMID 19618637	H5N1 highly pathogenic avian influenza in Southeast Asia	Eagles	2009
PMID 18598616	Toward a unified nomenclature system for highly pathogenic avian influenza virus (H5N1)	WHO/OIE/FAO	2008
PMID 24698181	Genomic analyses detect Eurasian-lineage H10 and additional H14 influenza A viruses recovered from waterfowl in the Central United States. Influenza Other Respir	Fries	2014
PMID 24647410	North Atlantic migratory bird flyways provide routes for intercontinental movement of avian influenza viruses	Dusek	2014
PMID 20933610	Evidence of intercontinental transfer of North American lineage avian influenza virus into Korea	Lee	2011
PMID 19995585	M., Ramey A. M., Ip H. S. & Gill R. E	Pearce	2010
PMID 22008513	Migratory flyway and geographical distance are barriers to the gene flow of influenza virus among North American birds	Lam	2012
PMID 19140989	V., Pearce J. M., Flint P. L., Franson J. C. & Ip H. S. Genetic evidence of intercontinental movement of avian influenza in a migratory bird: the northern pintail (Anas acuta)	Koehler	2008
PMID 25192767	Highly pathogenic avian influenza virus (H5N8) in domestic poultry and its relationship with migratory birds in South Korea during 2014	Jeong	2014
PMID 25597538	Comparing introduction to Europe of highly pathogenic avian influenza viruses A(H5N8) in 2014 and A(H5N1) in 2005	Adlhoch	2014
PMID 25898265	Novel Eurasian highly pathogenic avian influenza a H5 viruses in wild birds, Washington, USA, 2014	Ip	2015
PMID 25838478	Novel H5 Clade 2.3.4.4 Reassortant (H5N1) virus from a green-winged teal in Washington, USA	Torchetti	2015
PMID 25897841	Pathologic changes in wild birds infected with highly pathogenic avian influenza A (H5N8) viruses, South Korea, 2014	Kim	2015
PMID 26223637 In OmniVira	Pathogenesis and Transmission of Novel Highly Pathogenic Avian Influenza H5N2 and H5N8 Viruses in Ferrets and Mice.	Pulit-Penaloza	2015
PMID 19889765	A genetically engineered waterfowl influenza virus with a deletion in the stalk of the neuraminidase has increased virulence for chickens	Munier	2010
PMID 15890915	Improvement of influenza A/Fujian/411/02 (H3N2) virus growth in embryonated chicken eggs by balancing the hemagglutinin and neuraminidase activities, using reverse genetics	Lu	2005
PMID 10864641	Interdependence of hemagglutinin glycosylation and neuraminidase as regulators of influenza virus growth: a study by reverse genetics	Wagner	2000
PMID 8419645	Biologic importance of neuraminidase stalk length in influenza A virus	Castrucci M. R. & Kawaoka	1993
PMID 18779858	J., Hickman D. & Perez D. R. Evidence of expanded host range and mammalian-associated genetic changes in a duck H9N2 influenza virus following adaptation in quail and chickens	Hossain	2008
PMID 19225004	Neuraminidase Stalk Length and Additional Glycosylation of the Hemagglutinin Influence the Virulence of Influenza H5N1 Viruses for Mice	Matsuoka	2009
PMID 21373190	J., Miller J. & Carpenter T. Emergence and genetic variation of neuraminidase stalk deletions in avian influenza viruses	Li	2011
PMID 11044127	Molecular Correlates of Influenza A H5N1 Virus Pathogenesis in Mice	Katz	2000
PMID 23499672	Molecular signatures of virulence in the PB1-F2 proteins of H5N1 influenza viruses	Smith A. M. & McCullers	2013
PMID 26143617	Human infection with a novel, highly pathogenic avian influenza A (H5N6) virus: Virological and clinical findings	Pan	2015
PMID 26222578	F., Mok C. K., Peiris J. S. & Zhong N. S. Human infection with a novel avian influenza A (H5N6) virus. N. Engl	Yang	2015
PMID 16051873	Studies of H5N1 influenza virus infection of pigs by using viruses isolated in Vietnam and Thailand in 2004	Choi	2005
PMID 16502281	Pathogenicity of a highly pathogenic avian influenza virus, A/chicken/Yamaguchi/7/04 (H5N1) in different species of birds and mammals	Isoda	2006
PMID 26391278	PB2 protein of A(H7N9) influenza virus increased its virulence in mammalian models	Zhu	2015
PMID 15235128	The evolution of H5N1 influenza viruses in ducks in southern China	Chen	2004
PMID 24268805	Pathogenicity and transmission of H5N1 avian influenza viruses in different birds	Yuan	2014
PMID 12202562	Development of a real-time reverse transcription PCR assay for type A influenza virus and avian H5 and H7 hemagglutinin subtypes	Spackman	2002
PMID 11811679	Universal primer set for the full-length amplification of all influenza A viruses	Hoffmann	2001
PMID 8336541	Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA of humans and chimpanzees	Tamura K. & Nei	1993
PMID 24132122	MEGA6: Molecular evolutionary genetics analysis version 6.0	Tamura	2013
PMID 18006085	Development and application of monoclonal antibodies against avian influenza virus nucleoprotein	Yang	2008
-	Estimation of LD50 and its error by means of log-probit graph paper	Miller L. C. & Tainter	1944