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Reassortant highly pathogenic influenza A H5N2 virus containing gene segments related to Eurasian H5N8 in British Columbia, Canada, 2014.

Pasick J, Berhane Y, Joseph T, Bowes V, Hisanaga T, Handel K, Alexandersen S - Sci Rep (2015)

Bottom Line: In late November 2014 higher than normal death losses in a meat turkey and chicken broiler breeder farm in the Fraser Valley of British Columbia initiated a diagnostic investigation that led to the discovery of a novel reassortant highly pathogenic avian influenza (HPAI) H5N2 virus.Since its first appearance in Korea in January 2014, HPAI H5N8 spread to Western Europe in November 2014.These European outbreaks happened to temporally coincide with migratory waterfowl movements.

View Article: PubMed Central - PubMed

Affiliation: Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Winnipeg, Manitoba, Canada R3E 3M4.

ABSTRACT
In late November 2014 higher than normal death losses in a meat turkey and chicken broiler breeder farm in the Fraser Valley of British Columbia initiated a diagnostic investigation that led to the discovery of a novel reassortant highly pathogenic avian influenza (HPAI) H5N2 virus. This virus, composed of 5 gene segments (PB2, PA, HA, M and NS) related to Eurasian HPAI H5N8 and the remaining gene segments (PB1, NP and NA) related to North American lineage waterfowl viruses, represents the first HPAI outbreak in North American poultry due to a virus with Eurasian lineage genes. Since its first appearance in Korea in January 2014, HPAI H5N8 spread to Western Europe in November 2014. These European outbreaks happened to temporally coincide with migratory waterfowl movements. The fact that the British Columbia outbreaks also occurred at a time associated with increased migratory waterfowl activity along with reports by the USA of a wholly Eurasian H5N8 virus detected in wild birds in Washington State, strongly suggest that migratory waterfowl were responsible for bringing Eurasian H5N8 to North America where it subsequently reassorted with indigenous viruses.

No MeSH data available.


Related in: MedlinePlus

Phylogenetic characterization of the hemagglutinin genes of Canadian HPAI H5N2 virus isolates.The complete hemagglutinin (HA) gene sequences of the Canadian HPAI H5N2 viruses were aligned with H5 genes obtained from NCBI and GISAID EpiFlu™ databases using Clustal W. The supplementary file contains additional phylogenetic trees for the remaining gene segments and a Table that list and acknowledges the sequences included from GISAID. Phylogenetic and molecular evolutionary analysis was conducted using MEGA version 6 and the Maximum Likelihood method based on the Tamura Nei model28. The tree with the highest log likelihood is shown. Bootstrap test involved 500 replicates to determine reliability of the inferred tree with only bootstrap values above 70% shown. Black upright triangles, Canadian HPAI H5N2; inverted triangle single Canadian human case of HPAI H5N1. Clade designations are based on WHO/OIE/FAO H5N1 Evolution Working Group nomenclature (http://www.who.int/influenza/gisrs_laboratory/h5_nomenclature_clade2344/en/).
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f1: Phylogenetic characterization of the hemagglutinin genes of Canadian HPAI H5N2 virus isolates.The complete hemagglutinin (HA) gene sequences of the Canadian HPAI H5N2 viruses were aligned with H5 genes obtained from NCBI and GISAID EpiFlu™ databases using Clustal W. The supplementary file contains additional phylogenetic trees for the remaining gene segments and a Table that list and acknowledges the sequences included from GISAID. Phylogenetic and molecular evolutionary analysis was conducted using MEGA version 6 and the Maximum Likelihood method based on the Tamura Nei model28. The tree with the highest log likelihood is shown. Bootstrap test involved 500 replicates to determine reliability of the inferred tree with only bootstrap values above 70% shown. Black upright triangles, Canadian HPAI H5N2; inverted triangle single Canadian human case of HPAI H5N1. Clade designations are based on WHO/OIE/FAO H5N1 Evolution Working Group nomenclature (http://www.who.int/influenza/gisrs_laboratory/h5_nomenclature_clade2344/en/).

Mentions: On November 28, 2014, in Abbotsford, British Columbia, Canada a sudden increase in mortality was observed in a flock of 11,000 83-day old meat turkeys. The mortality rate would exceed 70% over the next 3 days. On November 30, 2014 a broiler breeder flock 8 km away in neighbouring Chilliwack, British Columbia experienced a10% mortality over a 24 hr period. Post-mortem examination of the chickens revealed lesions compatible with HPAI. These included severe facial edema, conjunctivitis, tracheal hyperemia, severe pulmonary congestion and edema and hemorrhages involving the mucosa of the proventriculus. Shank (tarsal) hemorrhages were also present. In contrast, the lesions in the turkeys were limited to varying degrees of generalized dark patchy congestion of the skeletal musculature and mild superficial congestion of the cecal tonsils. In both cases RNA extracted from tracheal swabs, cloacal swabs and tissues were positive for H5 subtype virus by real-time reverse transcription PCR. Using a universal primer set16 the full sequence of all 8 genes was obtained directly from diagnostic specimens and viral isolates from these two outbreaks. Based on BLAST searches (http://blast.ncbi.nlm.nih.gov/Blast.cgi) the closest gene sequences of the 8 viral gene segments in the NCBI database (Table 1) were identified which suggested the presence of a novel reassortant HPAI H5N2 virus that possessed gene segments related to Eurasian HPAI H5N8 viruses and North American lineage waterfowl viruses. The GenBank accession numbers for the HPAI H5N2 turkey isolate and the broiler isolates are KP307954 - KP307961 and KP795726 - KP795741. Phylogenetic analysis of the HA inferred a higher degree of relatedness to 2014 Korean H5N8 rather than 2014 UK, German and Dutch H5N8 isolates (Figure 1). It is important to note that the HA of the Canadian poultry H5N2 viruses was unrelated to the H5N1 that was associated with the human case in Alberta, Canada in 201317. Phylogenetic trees for the remaining gene segments are found in supplementary Figures 1 through 7 that also contains a Table which list and acknowledge the sequences included from GISAID. Viruses isolated from the chicken broiler breeder and meat turkey farms were essentially genetically identical and had intravenous pathogenicity indices of 2.98 and 2.96 respectively. The deduced amino acid sequence of the connecting peptide of the HA is LRERRRKR/GLF and identical to that reported for 2014 Korean HPAI H5N8 viruses. Amino acids E190, R220, G225, Q226 and G228 (H3 numbering) involved in receptor binding indicates that this virus has an affinity for α-2,3-linked glycans18 or avian specificity, however, amino acids A138 and A160, also present in this virus, have been previously reported to be associated with enhanced binding to α-2,6-linked glycans or human receptor specificity19. The NA, of typical North American wild bird origin, shows no stalk truncation which has been associated with adaptation to gallinaceous poultry20 suggesting a very recent introduction of this virus into poultry. The NA catalytic site residues R118, E119, D151, R152, W178, I222, R224, E227, A246, E276, R292, and R371 along with H274 indicate susceptibility to antiviral neuraminidase inhibitors, however the asparagine 31 mutation present in the M2 protein is associated with resistance to amantadine21. The virus has a glutamic acid instead of a lysine at amino acid residue 627 of the PB2 protein further indicating an avian adapted virus. In summary, this represents the first HPAI outbreak in North America that involves a virus with a Eurasian Gs/GD-lineage HA gene. Some of the viruses, such as H5N1 and H5N6, bearing the HA gene of this lineage have a proven zoonotic and possibly pandemic potential and for this reason a risk analysis for potential human infection should be carried out, the situation followed very closely and everything done to decisively control and eradicate the infection in poultry.


Reassortant highly pathogenic influenza A H5N2 virus containing gene segments related to Eurasian H5N8 in British Columbia, Canada, 2014.

Pasick J, Berhane Y, Joseph T, Bowes V, Hisanaga T, Handel K, Alexandersen S - Sci Rep (2015)

Phylogenetic characterization of the hemagglutinin genes of Canadian HPAI H5N2 virus isolates.The complete hemagglutinin (HA) gene sequences of the Canadian HPAI H5N2 viruses were aligned with H5 genes obtained from NCBI and GISAID EpiFlu™ databases using Clustal W. The supplementary file contains additional phylogenetic trees for the remaining gene segments and a Table that list and acknowledges the sequences included from GISAID. Phylogenetic and molecular evolutionary analysis was conducted using MEGA version 6 and the Maximum Likelihood method based on the Tamura Nei model28. The tree with the highest log likelihood is shown. Bootstrap test involved 500 replicates to determine reliability of the inferred tree with only bootstrap values above 70% shown. Black upright triangles, Canadian HPAI H5N2; inverted triangle single Canadian human case of HPAI H5N1. Clade designations are based on WHO/OIE/FAO H5N1 Evolution Working Group nomenclature (http://www.who.int/influenza/gisrs_laboratory/h5_nomenclature_clade2344/en/).
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Related In: Results  -  Collection

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f1: Phylogenetic characterization of the hemagglutinin genes of Canadian HPAI H5N2 virus isolates.The complete hemagglutinin (HA) gene sequences of the Canadian HPAI H5N2 viruses were aligned with H5 genes obtained from NCBI and GISAID EpiFlu™ databases using Clustal W. The supplementary file contains additional phylogenetic trees for the remaining gene segments and a Table that list and acknowledges the sequences included from GISAID. Phylogenetic and molecular evolutionary analysis was conducted using MEGA version 6 and the Maximum Likelihood method based on the Tamura Nei model28. The tree with the highest log likelihood is shown. Bootstrap test involved 500 replicates to determine reliability of the inferred tree with only bootstrap values above 70% shown. Black upright triangles, Canadian HPAI H5N2; inverted triangle single Canadian human case of HPAI H5N1. Clade designations are based on WHO/OIE/FAO H5N1 Evolution Working Group nomenclature (http://www.who.int/influenza/gisrs_laboratory/h5_nomenclature_clade2344/en/).
Mentions: On November 28, 2014, in Abbotsford, British Columbia, Canada a sudden increase in mortality was observed in a flock of 11,000 83-day old meat turkeys. The mortality rate would exceed 70% over the next 3 days. On November 30, 2014 a broiler breeder flock 8 km away in neighbouring Chilliwack, British Columbia experienced a10% mortality over a 24 hr period. Post-mortem examination of the chickens revealed lesions compatible with HPAI. These included severe facial edema, conjunctivitis, tracheal hyperemia, severe pulmonary congestion and edema and hemorrhages involving the mucosa of the proventriculus. Shank (tarsal) hemorrhages were also present. In contrast, the lesions in the turkeys were limited to varying degrees of generalized dark patchy congestion of the skeletal musculature and mild superficial congestion of the cecal tonsils. In both cases RNA extracted from tracheal swabs, cloacal swabs and tissues were positive for H5 subtype virus by real-time reverse transcription PCR. Using a universal primer set16 the full sequence of all 8 genes was obtained directly from diagnostic specimens and viral isolates from these two outbreaks. Based on BLAST searches (http://blast.ncbi.nlm.nih.gov/Blast.cgi) the closest gene sequences of the 8 viral gene segments in the NCBI database (Table 1) were identified which suggested the presence of a novel reassortant HPAI H5N2 virus that possessed gene segments related to Eurasian HPAI H5N8 viruses and North American lineage waterfowl viruses. The GenBank accession numbers for the HPAI H5N2 turkey isolate and the broiler isolates are KP307954 - KP307961 and KP795726 - KP795741. Phylogenetic analysis of the HA inferred a higher degree of relatedness to 2014 Korean H5N8 rather than 2014 UK, German and Dutch H5N8 isolates (Figure 1). It is important to note that the HA of the Canadian poultry H5N2 viruses was unrelated to the H5N1 that was associated with the human case in Alberta, Canada in 201317. Phylogenetic trees for the remaining gene segments are found in supplementary Figures 1 through 7 that also contains a Table which list and acknowledge the sequences included from GISAID. Viruses isolated from the chicken broiler breeder and meat turkey farms were essentially genetically identical and had intravenous pathogenicity indices of 2.98 and 2.96 respectively. The deduced amino acid sequence of the connecting peptide of the HA is LRERRRKR/GLF and identical to that reported for 2014 Korean HPAI H5N8 viruses. Amino acids E190, R220, G225, Q226 and G228 (H3 numbering) involved in receptor binding indicates that this virus has an affinity for α-2,3-linked glycans18 or avian specificity, however, amino acids A138 and A160, also present in this virus, have been previously reported to be associated with enhanced binding to α-2,6-linked glycans or human receptor specificity19. The NA, of typical North American wild bird origin, shows no stalk truncation which has been associated with adaptation to gallinaceous poultry20 suggesting a very recent introduction of this virus into poultry. The NA catalytic site residues R118, E119, D151, R152, W178, I222, R224, E227, A246, E276, R292, and R371 along with H274 indicate susceptibility to antiviral neuraminidase inhibitors, however the asparagine 31 mutation present in the M2 protein is associated with resistance to amantadine21. The virus has a glutamic acid instead of a lysine at amino acid residue 627 of the PB2 protein further indicating an avian adapted virus. In summary, this represents the first HPAI outbreak in North America that involves a virus with a Eurasian Gs/GD-lineage HA gene. Some of the viruses, such as H5N1 and H5N6, bearing the HA gene of this lineage have a proven zoonotic and possibly pandemic potential and for this reason a risk analysis for potential human infection should be carried out, the situation followed very closely and everything done to decisively control and eradicate the infection in poultry.

Bottom Line: In late November 2014 higher than normal death losses in a meat turkey and chicken broiler breeder farm in the Fraser Valley of British Columbia initiated a diagnostic investigation that led to the discovery of a novel reassortant highly pathogenic avian influenza (HPAI) H5N2 virus.Since its first appearance in Korea in January 2014, HPAI H5N8 spread to Western Europe in November 2014.These European outbreaks happened to temporally coincide with migratory waterfowl movements.

View Article: PubMed Central - PubMed

Affiliation: Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Winnipeg, Manitoba, Canada R3E 3M4.

ABSTRACT
In late November 2014 higher than normal death losses in a meat turkey and chicken broiler breeder farm in the Fraser Valley of British Columbia initiated a diagnostic investigation that led to the discovery of a novel reassortant highly pathogenic avian influenza (HPAI) H5N2 virus. This virus, composed of 5 gene segments (PB2, PA, HA, M and NS) related to Eurasian HPAI H5N8 and the remaining gene segments (PB1, NP and NA) related to North American lineage waterfowl viruses, represents the first HPAI outbreak in North American poultry due to a virus with Eurasian lineage genes. Since its first appearance in Korea in January 2014, HPAI H5N8 spread to Western Europe in November 2014. These European outbreaks happened to temporally coincide with migratory waterfowl movements. The fact that the British Columbia outbreaks also occurred at a time associated with increased migratory waterfowl activity along with reports by the USA of a wholly Eurasian H5N8 virus detected in wild birds in Washington State, strongly suggest that migratory waterfowl were responsible for bringing Eurasian H5N8 to North America where it subsequently reassorted with indigenous viruses.

No MeSH data available.


Related in: MedlinePlus