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Detection of
HPH5N8 in
Mute swans in
Sweden-2015
NATIONAL VETERINARY INSTITUTE
Siamak Zohari
Dep. of Microbiology
National Veterinary Institute, SVA
Siamak.Zohari@SVA.se
Courtesy to DN
Lead poisoning; 6-26 mg/kg body weight
20150305
Juvenile, male
20150220
Adult, male
20150305
Juvenile, female
20150305
Adult, male
20150218
Adult, male
Stockholm
Stockholm
Specific identification of influenza
virus
Classical
pathway
Virus isolation
Serological
characterization
Step 1
Detection of influenza A virus
Negative
Positive
Step 2
Molecular
pathway
M-gene qRT-PCR
Recommended by
EU-ref. lab.
Recommended
by EU-ref. lab.
Sequencing
of complete
genome
NegativeH5/H7/H9
qRT-PCR
Positive
Pathotyping
Step 3
Recommended
by EU-ref. lab.
Workflow for Detection of influenza A viruses
Phylogenetic tree of HA gene of H5-AIV
based on 1000bp of heamagglutinin (HA1)
gene. Phylogenetic tree was generated by
the NJ method as implemented in MEGA
software. The bootstrap test (2000 replicates)
are shown next to the branches
A turkey Germany MV R2472 2014
A chicken Netherlands emc 3 2014
A eurasian wigeon Netherlands emc 1 2014
A eurasian wigeon Netherlands emc 2 2014
A turkey Italy 14VIR7898 10 2014
A turkey Germany NI R3372 2014
A duck England 36254 14
A turkey Germany R2474 L00899 2014
A Chicken Netherlands 14015526 2014
A chicken Netherlands 14015531 2014
A MuteSwan Sweden SVA1503110277 SZ0502 2015
A MuteSwan Sweden SVA1503110141 SZ0543 2015
A duck England 36038 14
A duck England 36226 14
A wigeon Sakha 1 2014
Europe
2014-2015
A spot billed duck Korea H455 42 2014
A tundra swan Korea H411 2014
A Korean native chicken Korea H257 2014
A Baikal teal Korea H62 2014
A Coot Korea H81 2014
SouthKoria
2014
A American green winged teal Washington 195750 2014
A Northern pintail Washington 40964 2014
A gyrfalcon Washington 41088 6 2014
A turkey Washington 61 22 2014
A chicken Washington 61 9 2014
A domestic duck Washington 61 16 2014
A turkey BC FAV10 2014
NorthAmerica
2014-2015
Clade2.3.4.4
A duck Jiangsu k1203 2010
A herring gull Sweden V1116 06
A tufted duck Sweden V599 06
A mink Sweden V907 06
A goosander Sweden V539 06
A tufted duck Sweden V1027 06
A tufted duck Sweden V998 06
A eagle owl Sweden V1218 2006
A tufted duck Sweden V526 06
A eagle owl Sweden V618 2006
A Canada goose Sweden V978 06
A mute swan Sweden V827 06
A smew Sweden V820 06 Clade2.2&2.2.1
Asian-Hp-H5
A duck Ireland 113 1983
A mallard Sweden 49 2002
A mallard Sweden 40 2002
A mallard Sweden 64 2002
A mallard Sweden 21 2002
A mallard Sweden 101020 2009
A mallard Sweden 101224 2009
A mallard Sweden 74 2003
A mallard Sweden 7 2002
A mallard Sweden 39 2002
LP-EurasianH5
100
99
90
100
80
93
96
100
100
100
100
99
9893
88
88
94
0.01
Sweden2006
Communications
• Flu-lab net community
• Formal reporting requirements (OIE) completed
20th of March
• Sequence data available at the GISAID
database (EPI576389-EPI576396) 20th of March
• Samples shipped to APHA 13th of April
Single Species
2014-09-04
Multiple outbreaks
2014-01/2014-05/2014-07/2014-09
Multiple outbreaks
2014-04/ 2014-11/ 2014-12
Singel Species
2014-09
Multiplel outbreaks
2014-11-04
Multiplel Species
2014-11-14/2014-12-01
Single Species
2014-11-14
Multiple outbreaks
2014-12-10
Single outbreaks
2014-12-15
Single Species
2015-02-18/2015-03-05
Time line of major HP-H5N8 events
What would be
the opinion of
seal about this?
Index cases - Encountered out
in the sea, had difficulties to
"dive" / killed 20140416
Autopsied at NHR
20140429
The Swedish
Museum of Natural
History
C T C C A A T G G G T G C A A T A A A C T C C A G
A Swine Sweden U140218 0146 14VIR021694 2014
A Swine Sweden U140403 0142 14VIR041063 2014
A Swine Sweden U140415 0065 14VIR043722 2014
A Swine Sweden U140307 0144 14VIR030253 2014
A Swine Sweden U140320 0157 14VIR035318 2014
A Vinninga 14 05639
A Vinninga 14 06193
A Swine Sweden U140220 0029 14VIR22931 2014
A Swine Sweden U140206 0064 14VIR16416 2014
A Swine Sweden U140214 0119 14VIR020949 2014
Subclade IIb
A Swine Sweden U140212 0109 14VIR019417 2014
A Swine Sweden U140307 0152 14VIR030363 2014
A Swine Sweden U140211 0465 14VIR14040 2014
A Swine Sweden U140305 0126 14VIR028935 2014
A California 07 JRO09 2009 G1
Subclade IIa
Clade II
A Swine Sweden U140404 0141 14VIR041413 2014
A Swine Sweden U130506 D1070 2013
A Swine Sweden U130917 0131 13VIR131457 2013
A Swine Sweden U131022 0554 13VIR149894 2013
Clade I
94
95
93
97
93
94
93
0.005
20140617
Virological results
Neg-PDV
Neg-Pan herpes
Pos-Influenza A
H10
20140623
20140621
Outbreak of Influenza A virus in Northern Europe 2014
2014-04
2014-07
2014-08
2014-08
2014-09
2014-10
2014-10
A/domestic/duck/Republic/of/Georgia/2/2010/H10N7
A/domestic/duck/Republic/of/Georgia/1/2010/H10N7
A/mallard/Sweden/105536/2009/H10N1
A/mallard/Sweden/105501/2009/H10N1
A/mallard/Sweden/133546/2011/H10N4
A/Seal/Sweden/SVA0546/2014/H10N7
A/avian/Israel/543/2008/H10N7
A/Jiangxi/Donghu/346/2/2013/H10N8
A/Jiangxi/Donghu/346/1/2013/H10N8
Recent
A/chicken/Germany/N/1949
A/WhistlingSwan/Shimane/468/1988
A/swan/Shimane/1331/1981
A/Mink/Sweden/3900/1984/H10N4
A/Mallard/Gloucestershire/PD374/1985
A/Fowl/Hampshire/PD378/1985
Old
Eurasian/Lineage
A/Sydney/2/2010/H10N7
A/chicken/SYDNEY/809/2010
A/Chicken/NSW/Sydney/D891/2010
A/mallard/Wisconsin/11OS3578/2011
A/mallard/Arkansas/12OS184/2012
A/American/green/winged/teal/Mississippi/12OS405/2012
American/Lineage
H10
H7A/seal/Massachusetts/1/1980/H7N7
A/Caspian/seal/Russia/T1/2012/H4N6
A/Caspian/seal/Russia/1884/2002/H4N6
A/seal/Massachussetts/133/1982/H4N5
H4
A/harbor/seal/New/Hampshire/179629/2011/H3N8
A/harbor/seal/Massachusetts/1/2011/H3N8
A/seal/Massachusetts/3911/1992/H3N3
A/seal/Massachusetts/3984/1992/H3N3
H3
A/Elephantseal/California/2/2010/H1N1
A/Elephantseal/California/1/2010/H1N1
H1
100
100
100
100
100
100
100
100
100
99
100
100
100
100
99
92
90
100
100
100
100
100
100
94
99
0.05
HumanHuman
HA
Swedish Civil Contingencies Agency
Ministry of Defense
Roles and responsibilities
Ministry of
Enterprise and
Innovation
Ministry for
Health and Social
Affairs
Ministry of
Finance
NationalRegionalLocal
Administrativelevel
Board of Agriculture
National Veterinary
Institute
National Food
Agency
National Board of
Health and Welfare
Ministry for
Employment
Swedish Work
Environment
Authority
County
Administrative
Boards (n=21)
County councils
(n=20)
Municipalities
(n=290)
Swedish
Environmental
Protection Agency
Min. of the
Environment
and Energy
Swedish Agency for
Marine and Water
Management
Public Health
Agency of Sweden
The Swedish Dairy Association
Swedish Animal Health Service
The Swedish Egg and Poultry Association
Swedish Poultry Meat Association
Swedish Fish Health Control Program
• 1997: The Zoonosis Council is founded
• 2007-2009: Strategy and national plan for inter-
agency collaboration developed and adopted –
formal establishment of
”Zoonossamverkansgruppen” (group for
collaboration on severe/larger/”media-engaging”
zoonotic events)
• 2011-2013: Establishment of ”SUBU” (group for
collaboration on routine outbreak investigations)
Development of animal-public health
collaborations in Sweden
Collaborative functions
Zoonosis Council
”peace-time” forum with a strategic focus
SUBU – for continuous exchange of information
(primarily) regarding zoonotic outbreaks/incidents
Zoonossamverkansgrupp
crisis management function
November December Januari Februari Mars
Capacity building and preparedness; Development of broad
detection methods Using influenza virus as a model.
A Stockholm 20 2013
A Stockholm 16 2014
A Swine Sweden U140211 0465 2014
A Swine Sweden U140305 0126 2014
A Swine Sweden U140212 0109 2014
A Stockholm 1 2012
A Stockholm 2 2011
A Stockholm 45 2009
A Stockholm 13 2010
pH1N1
A Swine Sweden U140206 0064 2014
A Swine Sweden U140220 0029 2014
A Swine Sweden U140218 0146 2014
A Vinninga 10 05639 2014
A Vinninga 12 06193 2014
A Swine Sweden U140403 0142 2014
A Swine Sweden U140307 0144 2014
A Swine Sweden U140320 0157 2014
A Swine Sweden U140415 0065 2014
pH1N2
A Swine Sweden U140404 0141 2014
A Swine Sweden U130506 D1070 2013
A Swine Sweden U130917 0131 2013
A Swine Sweden U131022 0554 2013
pH1-2009like
A swine Lidkoeping 1193 2002 H1N1
A swine Sweden 1021 2009 avH1N2
A Swine Sweden U140506 0194 2014
A swine Sweden 9706 2010 H1N2
A Swine Sweden U120314 0158 2012
A Swine Sweden U130227 0266 2013
A Swine Sweden U140213 0057 2014
AvianLikeH1
98
94
99
100
97
96
88
92
91
100
86
0.05
A phylogenetic tree of the
nucleotide sequences of the
complete HA gene of the
swine influenza viruses
detected in Sweden
HA
pH1N1 pH1N1
pH1N2 pH1N2
pH1N1
avH1N2
pH1N2
pH1N2 pH1N2
avH1N2 avH1N2
Swedish Animal Health orgaisation
Participating Farmers
Colleagues from
public health agency
Charlotta Moraeus
Tero Härkönen
Colleagues from NHR
Aleksija Neimane
Caroline Bröjer
Colleagues from SVA
Colleagues from VIP-MDS
Colleagues from

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HPAIV H5N8 in Sweden

  • 1. Detection of HPH5N8 in Mute swans in Sweden-2015 NATIONAL VETERINARY INSTITUTE Siamak Zohari Dep. of Microbiology National Veterinary Institute, SVA Siamak.Zohari@SVA.se
  • 2.
  • 4. Lead poisoning; 6-26 mg/kg body weight
  • 5. 20150305 Juvenile, male 20150220 Adult, male 20150305 Juvenile, female 20150305 Adult, male 20150218 Adult, male Stockholm Stockholm
  • 6. Specific identification of influenza virus Classical pathway Virus isolation Serological characterization Step 1 Detection of influenza A virus Negative Positive Step 2 Molecular pathway M-gene qRT-PCR Recommended by EU-ref. lab. Recommended by EU-ref. lab. Sequencing of complete genome NegativeH5/H7/H9 qRT-PCR Positive Pathotyping Step 3 Recommended by EU-ref. lab. Workflow for Detection of influenza A viruses
  • 7. Phylogenetic tree of HA gene of H5-AIV based on 1000bp of heamagglutinin (HA1) gene. Phylogenetic tree was generated by the NJ method as implemented in MEGA software. The bootstrap test (2000 replicates) are shown next to the branches A turkey Germany MV R2472 2014 A chicken Netherlands emc 3 2014 A eurasian wigeon Netherlands emc 1 2014 A eurasian wigeon Netherlands emc 2 2014 A turkey Italy 14VIR7898 10 2014 A turkey Germany NI R3372 2014 A duck England 36254 14 A turkey Germany R2474 L00899 2014 A Chicken Netherlands 14015526 2014 A chicken Netherlands 14015531 2014 A MuteSwan Sweden SVA1503110277 SZ0502 2015 A MuteSwan Sweden SVA1503110141 SZ0543 2015 A duck England 36038 14 A duck England 36226 14 A wigeon Sakha 1 2014 Europe 2014-2015 A spot billed duck Korea H455 42 2014 A tundra swan Korea H411 2014 A Korean native chicken Korea H257 2014 A Baikal teal Korea H62 2014 A Coot Korea H81 2014 SouthKoria 2014 A American green winged teal Washington 195750 2014 A Northern pintail Washington 40964 2014 A gyrfalcon Washington 41088 6 2014 A turkey Washington 61 22 2014 A chicken Washington 61 9 2014 A domestic duck Washington 61 16 2014 A turkey BC FAV10 2014 NorthAmerica 2014-2015 Clade2.3.4.4 A duck Jiangsu k1203 2010 A herring gull Sweden V1116 06 A tufted duck Sweden V599 06 A mink Sweden V907 06 A goosander Sweden V539 06 A tufted duck Sweden V1027 06 A tufted duck Sweden V998 06 A eagle owl Sweden V1218 2006 A tufted duck Sweden V526 06 A eagle owl Sweden V618 2006 A Canada goose Sweden V978 06 A mute swan Sweden V827 06 A smew Sweden V820 06 Clade2.2&2.2.1 Asian-Hp-H5 A duck Ireland 113 1983 A mallard Sweden 49 2002 A mallard Sweden 40 2002 A mallard Sweden 64 2002 A mallard Sweden 21 2002 A mallard Sweden 101020 2009 A mallard Sweden 101224 2009 A mallard Sweden 74 2003 A mallard Sweden 7 2002 A mallard Sweden 39 2002 LP-EurasianH5 100 99 90 100 80 93 96 100 100 100 100 99 9893 88 88 94 0.01 Sweden2006
  • 8. Communications • Flu-lab net community • Formal reporting requirements (OIE) completed 20th of March • Sequence data available at the GISAID database (EPI576389-EPI576396) 20th of March • Samples shipped to APHA 13th of April
  • 9. Single Species 2014-09-04 Multiple outbreaks 2014-01/2014-05/2014-07/2014-09 Multiple outbreaks 2014-04/ 2014-11/ 2014-12 Singel Species 2014-09 Multiplel outbreaks 2014-11-04 Multiplel Species 2014-11-14/2014-12-01 Single Species 2014-11-14 Multiple outbreaks 2014-12-10 Single outbreaks 2014-12-15 Single Species 2015-02-18/2015-03-05 Time line of major HP-H5N8 events
  • 10.
  • 11. What would be the opinion of seal about this?
  • 12. Index cases - Encountered out in the sea, had difficulties to "dive" / killed 20140416 Autopsied at NHR 20140429 The Swedish Museum of Natural History C T C C A A T G G G T G C A A T A A A C T C C A G A Swine Sweden U140218 0146 14VIR021694 2014 A Swine Sweden U140403 0142 14VIR041063 2014 A Swine Sweden U140415 0065 14VIR043722 2014 A Swine Sweden U140307 0144 14VIR030253 2014 A Swine Sweden U140320 0157 14VIR035318 2014 A Vinninga 14 05639 A Vinninga 14 06193 A Swine Sweden U140220 0029 14VIR22931 2014 A Swine Sweden U140206 0064 14VIR16416 2014 A Swine Sweden U140214 0119 14VIR020949 2014 Subclade IIb A Swine Sweden U140212 0109 14VIR019417 2014 A Swine Sweden U140307 0152 14VIR030363 2014 A Swine Sweden U140211 0465 14VIR14040 2014 A Swine Sweden U140305 0126 14VIR028935 2014 A California 07 JRO09 2009 G1 Subclade IIa Clade II A Swine Sweden U140404 0141 14VIR041413 2014 A Swine Sweden U130506 D1070 2013 A Swine Sweden U130917 0131 13VIR131457 2013 A Swine Sweden U131022 0554 13VIR149894 2013 Clade I 94 95 93 97 93 94 93 0.005 20140617 Virological results Neg-PDV Neg-Pan herpes Pos-Influenza A H10 20140623 20140621
  • 13. Outbreak of Influenza A virus in Northern Europe 2014 2014-04 2014-07 2014-08 2014-08 2014-09 2014-10 2014-10
  • 14. A/domestic/duck/Republic/of/Georgia/2/2010/H10N7 A/domestic/duck/Republic/of/Georgia/1/2010/H10N7 A/mallard/Sweden/105536/2009/H10N1 A/mallard/Sweden/105501/2009/H10N1 A/mallard/Sweden/133546/2011/H10N4 A/Seal/Sweden/SVA0546/2014/H10N7 A/avian/Israel/543/2008/H10N7 A/Jiangxi/Donghu/346/2/2013/H10N8 A/Jiangxi/Donghu/346/1/2013/H10N8 Recent A/chicken/Germany/N/1949 A/WhistlingSwan/Shimane/468/1988 A/swan/Shimane/1331/1981 A/Mink/Sweden/3900/1984/H10N4 A/Mallard/Gloucestershire/PD374/1985 A/Fowl/Hampshire/PD378/1985 Old Eurasian/Lineage A/Sydney/2/2010/H10N7 A/chicken/SYDNEY/809/2010 A/Chicken/NSW/Sydney/D891/2010 A/mallard/Wisconsin/11OS3578/2011 A/mallard/Arkansas/12OS184/2012 A/American/green/winged/teal/Mississippi/12OS405/2012 American/Lineage H10 H7A/seal/Massachusetts/1/1980/H7N7 A/Caspian/seal/Russia/T1/2012/H4N6 A/Caspian/seal/Russia/1884/2002/H4N6 A/seal/Massachussetts/133/1982/H4N5 H4 A/harbor/seal/New/Hampshire/179629/2011/H3N8 A/harbor/seal/Massachusetts/1/2011/H3N8 A/seal/Massachusetts/3911/1992/H3N3 A/seal/Massachusetts/3984/1992/H3N3 H3 A/Elephantseal/California/2/2010/H1N1 A/Elephantseal/California/1/2010/H1N1 H1 100 100 100 100 100 100 100 100 100 99 100 100 100 100 99 92 90 100 100 100 100 100 100 94 99 0.05 HumanHuman HA
  • 15. Swedish Civil Contingencies Agency Ministry of Defense Roles and responsibilities Ministry of Enterprise and Innovation Ministry for Health and Social Affairs Ministry of Finance NationalRegionalLocal Administrativelevel Board of Agriculture National Veterinary Institute National Food Agency National Board of Health and Welfare Ministry for Employment Swedish Work Environment Authority County Administrative Boards (n=21) County councils (n=20) Municipalities (n=290) Swedish Environmental Protection Agency Min. of the Environment and Energy Swedish Agency for Marine and Water Management Public Health Agency of Sweden The Swedish Dairy Association Swedish Animal Health Service The Swedish Egg and Poultry Association Swedish Poultry Meat Association Swedish Fish Health Control Program
  • 16. • 1997: The Zoonosis Council is founded • 2007-2009: Strategy and national plan for inter- agency collaboration developed and adopted – formal establishment of ”Zoonossamverkansgruppen” (group for collaboration on severe/larger/”media-engaging” zoonotic events) • 2011-2013: Establishment of ”SUBU” (group for collaboration on routine outbreak investigations) Development of animal-public health collaborations in Sweden
  • 17. Collaborative functions Zoonosis Council ”peace-time” forum with a strategic focus SUBU – for continuous exchange of information (primarily) regarding zoonotic outbreaks/incidents Zoonossamverkansgrupp crisis management function
  • 18. November December Januari Februari Mars Capacity building and preparedness; Development of broad detection methods Using influenza virus as a model.
  • 19. A Stockholm 20 2013 A Stockholm 16 2014 A Swine Sweden U140211 0465 2014 A Swine Sweden U140305 0126 2014 A Swine Sweden U140212 0109 2014 A Stockholm 1 2012 A Stockholm 2 2011 A Stockholm 45 2009 A Stockholm 13 2010 pH1N1 A Swine Sweden U140206 0064 2014 A Swine Sweden U140220 0029 2014 A Swine Sweden U140218 0146 2014 A Vinninga 10 05639 2014 A Vinninga 12 06193 2014 A Swine Sweden U140403 0142 2014 A Swine Sweden U140307 0144 2014 A Swine Sweden U140320 0157 2014 A Swine Sweden U140415 0065 2014 pH1N2 A Swine Sweden U140404 0141 2014 A Swine Sweden U130506 D1070 2013 A Swine Sweden U130917 0131 2013 A Swine Sweden U131022 0554 2013 pH1-2009like A swine Lidkoeping 1193 2002 H1N1 A swine Sweden 1021 2009 avH1N2 A Swine Sweden U140506 0194 2014 A swine Sweden 9706 2010 H1N2 A Swine Sweden U120314 0158 2012 A Swine Sweden U130227 0266 2013 A Swine Sweden U140213 0057 2014 AvianLikeH1 98 94 99 100 97 96 88 92 91 100 86 0.05 A phylogenetic tree of the nucleotide sequences of the complete HA gene of the swine influenza viruses detected in Sweden HA pH1N1 pH1N1 pH1N2 pH1N2 pH1N1 avH1N2 pH1N2 pH1N2 pH1N2 avH1N2 avH1N2
  • 20. Swedish Animal Health orgaisation Participating Farmers Colleagues from public health agency Charlotta Moraeus Tero Härkönen Colleagues from NHR Aleksija Neimane Caroline Bröjer Colleagues from SVA Colleagues from VIP-MDS Colleagues from