Lecture 1, 2 by Kida Core Curriculum for Zoonosis Control 2010 Hokkaido University, Sapporo, Japan September p 2, , 2011

A i Avian, swine, swine i , and d pandemic influenza

Hiroshi Kida Professor, Graduate School of Veterinary Medicine Director, Research Center for Zoonosis Control Head, Head OIE Reference Laboratory for Avian Influenza Hokkaido University, Sapporo, Japan

How should we control avian influenza ? Are we substantially prepared for pandemic influenza ?
Why have the H5N1 HPAIVs persisted in poultry for 15 years ? Why Wh are antigenic ti i variants i t selected l t di in bi birds d ? Will the HPAIVs returned to migratory birds persist in nature ? How should avian influenza be controlled in poultry ? What are the advantage and disadvantage of the use of vaccines ? Is it OK to forget g about H5N1 avian influenza virus strain after the swine-origin H1N1 pandemic strain emerged ? Are the measures for the control of seasonal flu satisfactory ?

Ecology of influenza viruses in nature, birds and mammals;

Origin, Origin evolution, evolution and perpetuation of influenza virus in nature. Mechanism of the emergence of highly pathogenic avian flu virus and human pandemic strains. Antigenic variation and genetic reassortment of influenza viruses.

Influenza virus particle

Hemagglutinin HA, HA H1 H1-16 16 subtypes Neuraminidase NA, N1-9 subtypes Envelope

Infection, multiplication, and release of influenza virus

Fusion of viral envelope with the endosomal membrane Uncoating; penetration Internalization in Endosome 10 Formation of viral 10. particle Recepter binding

Viropexis

. Replication Transcription T i ti Translation Post translational modification 12. Release 11. Budding

Pig g
Cell
Genetic reassortment A/duck/S China/x/67

(H3NX)

A/S China/x/67

(H2N2)

A/Hong Kong/1/68

(H3N2)

H mans Humans

Pandemic influenza in humans

Swine-origin H1N1 pandemic strain

Highly pathogenic avian influenza

Derivation of the genes of pandemic influenza viruses

Subtype
Year

Number of the genes 3 4 5 6

H1N1
(1947)

H2N2
(1957)

H3N2
(1968)

PB2 PB1

PA HA NP NA Proteins encoded by the genes M NS

Gene derived from non-human virus

Host range, and HA and NA subtypes of influenza A virus

H1N1 H1N2 H3N2 H1N1 H2N2 H3N2 (H2N8,H3N8) H5N1 H7N7 H5N1,H7N7, H9N2

H2N3, H3N1, H3N3, H3N8 H4N6, H3N8, H4N6 H5N1, H5N1 H5N2, H9N2

H3N8 H7N7

H1-H12 N1 9 N1-9

H3N8,H5N1 H5N1

H1-10 N1-9

H1-16 N1-9 N1 9
H1-7, H9-16 N1-9 H3N3 H4N5 H7N7 H3N2 H5N1 H10N4 H1N3, H13N2, H13N9
8

H1-7, H9-11 N1-4, N6-8

Intestinal replication of AIV in duck

Tissue
B i Brain Pharynx Larynx Trachea, Lungs Esophagus,Proventriculus Duodenum, Jejunum Ileum

Virus recovery (log EID50)

-

FA
-

2.8 1.8
-

Cecum

Colon Rectum Bursa

, Liver, , Kidney, y, Pancreas, Genitals, Heart, Blood, Spleen, Thymus

3.5 3 5 5.0 8.0 7.5 6.0

-

+++ +
-

Feces

7.7
Kida et al (1980) Infect Immun

Duck infected with A/duck/Hokkaido/5/77 (H3N2)

D k influenza Duck i fl

Each of the known subtypes (H1-16, (H1 16 N1 N1-9) 9) of influenza A virus has been isolated from ducks. In d I ducks, k viruses i replicate li t i in th the colon, l b being i shed h d with feces in a week, and non-pathogenic. W t b Water-borne f fecal-oral l l transmission t i i Ducks carry and provide viruses during migration and over-wintering. i t i Migratory duck is the NATURAL HOST of influenza A viruses.
Kida et al (1980) Infect Immun

Wilson et al (1981) Nature

Neutralization of infectivity of influenza virus by anti-HA anti HA antibodies

IgG antibody HA HA HA IgG ab

Hemagglutination-inhibition (Receptor-binding-inhibition) Neutralization of viral infectivity

+ +

Reactivity of duck and human virus strains with the McAbs Virus strains
Duck Strains Dk/Ukraine/63 Dk/Hok/5/77 / / / Dk/Hok/8/80 Dk/Hok/33/80 Dk/Hok/7/82 Dk/Hok/21/82 Dk/Hok/9/85 / / / Dk/Hok/10/85 Human Strains Aichi/2/68 / / Ud orn/307/72 Tokyo/6/73 P Chalmers/1/73 P.Chalmers/1/73 Victoria/3/75 Kumamoto/22/76 Bangkok/1/79 Philipines/2/82

Monoclonal antibodies
13/1 110/2 48/2 22/1 32/2 D11/3 D13/1 D17/4 D58/2 D22/3

Nucleotide and amino acid sequence identity of theHAs of duck strains with that of A/Hong Kong/68
Virus strains
D k/Ho k/5/77 D k/Ho k/8/80 D k/Ho k/33/80 D k/H o k/7/82 D k/Ho k/9/85 D k/Ho k/10/85 / k/21/82 /2 /82 D k/Ho D k/Ukr/63 Aich i/2/68 N u cleo tid e seq u en ce id en tity ( %) D k/5/ D k/8/ D k/33 D k/7/ D k/9/ D k/10 D k/21 D k/Ukr Aich i/ 77 80 /80 82 85 /85 /82 /63 2/68 94 6 94.6 97.5 98.2 97 6 97.6 97.8 98.0 9 3 95.3 95.8 97.1 98.5 98 9 98.9 97.8 98.0 96 0 96.0 96.7 97.3 97.1 97 1 95.8 94.2 94 2 98.2 95.3 96.4 96 4 94.6 98.5 94 4 94.4 96.7 96 7 94.9 98.5 94 7 94.7 99.4 84.8 84 8 85.0 85.6 85 2 85.2 85.0 85.5 90.5 90 5 91.0 91.4 91 4 91.4 91.0 90.8 8 6 85.6 95.8 96.9 96 9 95.9 97.5 95 4 95.4 96.7 97.1 8 0 85.0 91.2

98.7 98 7 99.3 98.0 99.1 98.2 96 96.4 96 2 96.2 96.9 96.7 97.8 97.6 Amin o a cid

99.8 9 6 95.6 9 8 95.8 96.2 96.4 95.1 97.6 97.8 95.3 seq u en ce id en tity ( %)

H3 HAs
Amino acid substitutions on the HAs of viruses isolated from humans are confined to the antigenic sites, indicating that viruses are circulating in the presence of antibody selection pressure. Amino acid substitutions on the HAs of viruses isolated from ducks are scattered throughout the HA molecule and not confined to the antigenic i sites, i indicating that viruses are circulating in the absence of antibody selection pressure.

Extensive antigenic variation

Evolutionally stasis

Influenza f viruses are antigenically and genetically g y highly g y conserved in ducks.

Viruses replicating Vi li ti i in the th i intestine t ti are not t under d the serum antibody selection pressure. 40 to 50 % of population of migratory ducks are juvenile birds that hatched in the summer. Preserved in frozen water of the lakes, , where they y nest in summer, in winter.
Kida H et al (1987) Virology; Ito et al (1995) Arch Virol

Reactivity of swine, duck, and human strains with McAbs

Virus strains Sw/TW/125/82 S /TW/125/82 Sw/TW/126/82 Sw/TW/127/82 Sw/HK/81/78 Dk/Hok/8/80 Dk/Hok/7/82 Aichi/2/68 Victoria/3/75 13/1 110/2 48/2 Monoclonal antibodies 22/1 32/2 D11/3 D13/1 D17/4 D58/2 D22/3

Amino acid sequences at the receptor-binding site of the HA of swine viruses compared to those of human and avian strains Virus us st strain a sequence 6 226 Human Viruses Avian Viruses Sw/Taiwan/82 Sw/China/78 227 Amino Acid 228 8

Leu - Ser - Ser Gln Gln - Ser - Gly - Ser - Gly

Leu - Ser - Ser

Virus HA
NeuAc

Receptor
Gal
CHOH AcHN OH OH CH2OH HO O COOH O HO a O O OH CH2OH AcHN OH O COOH HO OH CH2OH O HO O O OH

GlcNAc
CH2OH

O O

OH NHAc

- - Leu Ser - Ser

2, 6

CH2OH O O OH NHAc

- - Gln - Ser - Gly

2, 3

Amino A i acid id residues id 226 and d 228 of f th the HA1 subunit b it of f H3 HA determine the receptor specificity.

Ito et al (1998) J Virol

HAs of H3N2 viruses isolated from pigs in southern China

Closely related with those of A/Hong Kong/68 (H3N2) and H3 viruses isolated from migratory ducks. Amino A i acid id sequences at t th the receptor-binding t bi di site it on the HA indicate either specificity to human or avian type receptors. receptors Both types yp of receptor p were found on the surface of the epithelial cells lining the upper respiratory tract of pigs.
KIDA et al (1988) Virology; Ito et al (1998) J Virol

Nucleotide N l tid and d amino i acid id sequence identity id tit of f H3 viruses i of f ducks, goose, and pig from China, duck from Japan, and A/Hong Kong/68 and its variant
Nucleotide sequence identity % Dk/HK Gs/HK Dk/HK Dk/HK/ Sw/HK/ Dk/Hok Aichi /7/75 /10/76 /64/76 231/77 126/82 /8/80 /2/68 96.6 97.1 92.5 99.0 98.0 95.6 97 5 97.5 99 0 99.0 93 5 93.5 96 7 96.7 96 2 96.2 95 2 95.2 97.8 98.9 93.2 97.3 96.7 95.4 96.6 97.3 96.9 93.2 92.7 94.0 99.5 97.6 98.0 96.9 98.3 96.5 97.8 97.5 97.8 96.7 98.7 95.9 96.9 96.9 96.9 96.9 97.1 97.3 94.0 93.6 93.6 93.1 94.6 94.4 95.6 Amono acid sequence identity (%) Vic/ 3/75 93.0 92 9 92.9 93.2 91.5 93.3 93.2 96.5

Dk/HK/7/75 G /HK/10/76 Gs/HK/10/76 Dk/HK/64/76 Dk/HK/231/77 Sw/TW/126/82 / / / Dk/Hok/8/80 Aichi/2/68 Vic/3/75

Yasuda et al (1991) J Gen Virol

Influenza viruses isolated from domestic ducks in southern China

HAs of influenza viruses isolated from domestic ducks in southern China closely y related with those of viruses isolated from migratory ducks, pigs, and A/Hong Kong /68 antigenically and genetically. Each of the viruses was isolated on the Pacific Flyway of migratory ducks. ducks Migratory duck domestic duck pig humans

Yasuda et al (1991) J Gen Virol

Pig g
Cell
Genetic reassortment A/duck/S China/x/67

(H3NX)

A/S China/x/67

(H2N2)

Domestic duck or goose g

A/Hong Kong/1/68

(H3N2)

H mans Humans

Migratory duck

A/duck/ xx (H3N?)

Domestic duck

Pond Virus shedding A/Asian/67 (H2N2)

Genetic G ti reassortment 1918 Spanish virus (H1N1)

1968 HK A/HongKong/68 g g (H3N2)

1957 Asian virus H2N2 must have appeared similarly.

Rout of transmission of the genes of pandemic strains

Kida et al (1994) J Gen Virol

3 4 10 17 6 7 Dk/8/80 Sw/2/81 () (H3N8) (H3N8) (H3N8) (H3N8) (H3N1) (H3N1) (H3N8) (H1N1) PB2 D D D D D D D S PB1 D D D D D D D S PA D D D D D D D S HA D D D D D D D S NP D D D D S S D S NA D D D D S S D S M D D D D S D D S NS D D D D D S D S

D A/d D: A/duck/Hokkaido/8/80 k/H kk id /8/80 (H3N8)

S A/ S: A/swine/Hokkaido/2/81 i /H kk id /2/81 (H1N1)

Kida et al (1994) J Gen Virol

The role of pigs in the emergence of pandemic influenza virus strains p

Pigs are susceptible to avian influenza viruses of each of the HA subtypes. Genetic reassortants were generated in the cells lining g upper pp respiratory p y tract of pig p g upon p concurrent infection with mammalian and avian strains.

Kida H et al (1994) J Gen Virol

Candidates of future pandemic strain

H1 to H16 and N1 to N9 subtypes of influenza A viruses perpetuate p p in lakes where ducks nest in nature. 1957 H2N2 and 1968 H3N2 viruses are reassortants between AIV and the preceding human strains. Pigs are susceptible to each of avian and mammalian influenza viruses, generating reassortants.

Avian viruses of any subtype can contribute genes for reassortants : None of the 16 HA and 9 NA subtypes can be ruled out as potential candidates for future pandemics. Global surveillance of swine flu as well as avian flu is i important. t t

Ito T et al (1995) Arch Virol

Isolation of influenza viruses from water samples of lakes in Alaska in 1992-1994 No. of samples with virus/ total no. of samples tested 1992 1993 1994 1994 summer summer summer autumn 1/4 0/2 0/3 0/3 0/4 0/1 0/4 0/5 7/13 3/21 0/10 1/2 0/5 0/17 0/5 0/3 0/1 2/23 0/28 7/30 3/21

Location
Lake Ho od /Spenard Lake Cheney P tt Potter M Marsh h Westchester Lagoon Lake Hanger Fairb anks

Big Minto Lake Mallard Lake

Heart Lake Canvasb ack Lake C Corville ill D Delta lt Total

Kobyaysky(820) 40 Ilands(1321) Kenkeme(32) H4N6 H4N9 H11N1 H11N6 H11N9 White Lake(1136) H3N8 Ptropavlovsk PtropavlovskKamchatsky(58)

Buotama(51)

Magadan(295)

Kh Kharyyalah(146) l h(146) Yakutsk(232) H3N8 H13N6 Khabarovsk(23)

Irkutsk(290)

Elavga(66)

L k K Lake Kanicheva(95) i h (95) Wakkanai (958) H1N1 H3N8 H5N3 H5N4 H6N2 H6N7 H8N1 H8N3 H9N2 H11N9

Malyshevo(90)

Taiwan H1N1 H4N5 H4N6 H7N7

1996 1997 1998 1999

Phylogenic tree of the HA genes Eurasian E i li lineage American lineage

Acquisition of pathogenicity of avian influenza virus in chicken

APAIV

69 Months

LPAIV

HPAIV (H5 or H7)

Subtype H1 H2 H3 H4 H5 H5 H6 H7 H7 H8 H9 9 H10 H11 H12 H13 H14 H15

Amino acid sequences at the cleavage site of influenza A virus HAs

Strains Dk/Alberta/35/76(H1N1)b Mal/MT/Y61(H2N2)b Dk/Menphis/928/74(H3N8)b Dk/Czechoslovakia/56(H4N6) ( )b Ck/Scotland/59(H5N1)b Ty/MN/3/92(H5N2)a Shw/Australia/1/72(H6N5)b FPV/Rostock/34(H7N1)b Mal/Alberta/195/89(H7N3)a Ty/Ontario/6118/68(H8N4)b Ty/Wisconsin/66(H9N2) / /66( 9 2)b Ck/Germany/N/49(H10N7)b Dk/England/56(H11N6)b Dk/Alb t /60/76(H12N5)b Dk/Alberta/60/76(H12N5) Gl/Maryland/704/77(H13N6)b Mal/Gurjev/263/82(H14N5)b Shw/Australia/2576/79(H15N9)b
a

A A sequences

IQSR IESR KQTR KASR RKKR RETR IETR KKRKKR KKTR VEPR RSSR VQGR IASR VQDR ISNR KQAK IRTR
b Kovacova

GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF GLF

Senne et al (1996),

et al (2002)

Return of the HPAIV from domestic poultry to migratory water birds

APAIV

> 6-9 months

LPAIV HPAIV (H5 or H7)

HPAI viruses isolated from wild birds in Mongolia

A/whooper swan/Mongolia/3/05 (H5N1) A/bar-headed goose/Mongolia/1/05 (H5N1) A/common goldeneye/Mongolia/12/06 (H5N1)

Ugii Lake

A/whooper swan/Mongolia/2/06 (H5N1)

Qinghai Lake

A/whooper swan/Mongolia/2/09 (H5N1) A/whooper swan/Mongolia/9/09 (H5N1) A/bar-headed goose/Mongolia/X53/09 (H5N1) A/ bb sholduck/Mongolia/X42/2009 A/rubby h ld k/M li /X42/2009 (H5N1) A/common goldeneye/Mongolia/X60/09 (H5N1) A/whooper swan/Mongolia/1/10 (H5N1) A/whooper swan/Mongolia/7/10 (H5N1)

62 Countries where H5N1 HPAIV infections were reported in wild birds, poultry, and both
Japan, Republic of Korea, China, Mongolia, Myanmar, Lao PDR, Thailand, Cambodia, Viet Nam, Malaysia, Indonesia, Bangladesh, India, Pakistan; Afghanistan, Afghanistan Iran, Iran Azerbaijan, Azerbaijan Georgia, Georgia Iraq, Iraq Kuwait, Kuwait Saudi Arabia, Arabia Turkey, Turkey Israel; Russian Federation, Federation Kazakhstan, Kazakhstan Ukraine Ukraine, Romania, Bulgaria, Albania, Serbia, Hungary, Slovakia, Czech Republic, Croatia, Poland, Slovenia, Bosnia & Herzegovina; Greece, Switzerland, Austria, France, Italy, Germany, Netherlands, Denmark, Sweden, Spain, England, Ireland; Djibouti, Gaza Strip, Egypt, Sudan, Nigeria, Niger, Cameroon, Burkina Faso, Cote dIvoire

Confirmed human cases of H5N1 HPAIV infection

Country Deaths /Cases 26 / 40 59 / 119 146 / 178 52 / 151 16 / 18 2 / 2 17 / 25 2 / 3 5 / 8 4 / 12 0 / 1 1 / 1 0 / 1 1 / 3 0 / 3 331 / 565 04 05 06 07 04 05 06 07 08 08 09 10 09 10 11 03 03 11
Chi China E Egypt

China Viet Nam Indonesia Egypt Cambodia Lao PDR Thailand Iraq Azerbaijan Turkey Djibouti Nigeria Myanmar Pakistan Bangladesh Total

Viet Nam

Thailand

Indonesia

Cambodia

As of 19 August 2011

WHO, Kuribayashi

Birdfluvaccines
Vietnam:
H5N2andH5N1 (Adjuvantinactivatedvaccines)

As a stockpile, Singapore:
H5N2 (Inactivated,adjuvanted vaccine)

China:
H5N1andrecombinantNDV (Reversegeneticsinactivatedvaccines)

Japan:
H5N1andH7N7 (Oiladjuvanted inactivated i ) vaccines)

Indonesia:
H5N1,H5N2,H5N9andrecombinant H5N1 (inactivatedvaccines)

Pakistan:
H5N1,H5N2,H5N9,andH5N3 (Waterbasedwithalumhydroxide andoilbasedwithmineraloil)

Egypt: since2006
Thailand: Prohibitedvaccine2006

Q.WhydoyouthinkthattheH5N1HPAIVstrainshavepersistedinpoultryfor12years? The virus strains are circulated among poultry populations in some Asian countries b because of f vaccination i ti . If the th vaccine i is i used, d infected i f t d poultry lt may not t be b recognized i d because they may not show any clinical signs. Eventually the poultry can be resources of the infection. Continuing spread of virus within and between large populations of susceptible birds, influenced by breeding and movement of birds, virus exposure and vaccination history, antigenic drift etc. etc.

Q.Why Q ydothesestrainsshowantigenic g variation? In some Asian countries, poultry are vaccinated. The antigenic variants should have been selected by the pressure of antibodies induced by vaccination. Theuseofvaccine, mixed poultry in a pen, poor managementofsanitation.

Influenza Vaccine for bird flu

may prevent manifestation of disease signs and decrease the amount of virus shed, but does not confer protective immunity from infection. Stamping-out p g influenza.

policy p y is recommended for the control of avian

Vaccination is not primarily y recommended but approved as one of the options applied only under DIVA (differentiating infected from vaccinated animals) based strategy. Country where vaccine is used is not designated as HPAI-free.

must lead silent spread of virus.

26TH CONFERENCE OF THE OIE REGIONAL COMMISSION FOR ASIA, THE FAR EAST AND OCEANIA Shanghai, Peoples Republic of China, 16 16-20 November 2009

RECOMMENDATION FOR THE CONTROL OF AVIAN INFLUENZA It is considered that;

Highly pathogenic avian influenza H5N1 virus strains have persisted in domestic poultry for 14 years and antigenic variants have been selected mainly due to the misuse of vaccine. HPAI has been put under control in several countries. Stamping out policy has been the most effective measures for the control HPAI. Vaccine is used in 4 countries where HPAI has not been controlled yet. Vaccine is used instead of stamping out in 2 countries and in the other 2 countries, in addition to stamping out. Sentinel bids are put in the vaccinated poultry population in Viet Nam and not in the other 3 countries where vaccine is used. Compensation for livestock owners is done in most countries in case of stamping out.

It is recommended that;
Since stamping out is the best and ultimate measure for the control of HPAI HPAI, vaccine should be used in addition to, not instead of stamping out. The OIE should continue and develop standards on animal influenza surveillance, prevention and control. Surveillance of swine flu is crucial in the countries where avian flu has not been controlled.

Global surveillance of avian influenza in autumn (19912009)

Mongolia (324) H1N1 (5) H2N2 (1) H2N3 (1) H3N2 (2) H3N6 (34) H3N8 (149) H4N2 (2) H4N3 (1) H4N6 (72) H4N7 (1) H4N8 (4) H5N2 (1) H5N3 (4) H7N1 (2) H7N6 (1) H7N7 (13) H7N9 (3) H8N4 (6) H9N2 (1) H10N3 (13) H10N5 (2) H10N7 (5) H12N5 (1) Russia (56) H3N8 (17) H4N9 (2) H11N1 (1) H11N9 (8) H4N6 (25) H10N7 (1) H11N6 (1) H13N6 (1)

USA (111) H2N3 (1) H4N6 (57) H7N7 (1) H10N7 (11) ( )

H3N8 (39) H7N3 (1) H8N2 (1)

China (2) H3N8 (1) H4N6 (1)

Australia (6) H2N5 (6)

Hokkaido (296) H1N1 (12) H2N2 (2) H2N5 (1) H3N2 (2) H3N8 (37) H4N2 (8) H4N6 (33) H4N9 (3) H5N3 (11) ( ) H6N1 (17) ( ) H6N5 (2) H6N8 (7) H7N1 (18) H7N7 (14) H9N2 (7) H9N4 (1) H9N9 (1) H10N2 (1) H10N5 (7) H10N6 (1) H10N8 (1) H10N9 (2) H11N9 (21) H12N2 (2) H13N6 (2) H5N1(1)

H2N3 (4) H3N6 (6) H4N5 (1) H5N2 (1) H6N2 (32) ( ) H6N9 (1) H8N4 (8) H9N5 (1) H10N4 (12) H10N7 (12) H11N6 (1) H12N5 (4)

795 isolates from 22,744 samples

Surveillance of avian influenza in autumn 2010

Mongolia(36isolates) H1N1(1)H3N3(1) H3N6(7)H3N8(14) H4N6(8)H7N9(1) H10N8(4) Hokkaido(15isolates) H3N8(3)H5N2(1) H6N2(2)H7N7(9)

H5N1(2)
HongKong(3isolates) H3N2(1) H5N1(2)

Vietnam(1isolate) H9N6(1)

Laos(none)

Number of samples 4,515 Influenza virus isolates 55 As of 25 October 2010

H5N1 HPAIVs isolated from wild birds and poultry in Japan, 2010-2011
October, 2010 Fecal samples of ducks Wakkanai, Hokkaido

Poultry Wild birds Other

, February, 2011 Chickens (layer) (2 farm farm, 327 327,000) 000) Mie February, 2011 Chickens (broiler)(1 farm, 100,000) G j Nara Gojo, N November, 2010 Chickens (layer) (1 farm, 23,000) Y Yasugi, i Shimane Shi

January, 2011 February, 2011 Chickens (layer) (2 farm, 166,000) Aichi

February, 2011 Chickens (layer) (1 farm farm, 8 8,100) 100) Oita, Oita ,, January, 2011 February, 2011 ,, March, 2011 Chickens (13 farm, 1,012,000) Miyazaki

, March, 2011 Chickens (layer, Broiler)(2 farm, 97,000) Chiba, Chiba

January, January 2011 Chickens (layer) (1 farm, 8,600) Izumi, Kagoshima

February, 2011 Chickens (broiler)(1 farm, 100,000) Kinokawa, Wakayama

11 April 2011, Okamatsu, MAFF

H5N1 HPAIV infections in Korea since the end of December 2010 as of 18 May 2011
12017 21318 127 1 1 27 29135 18153 123134 573 1231117 1251100 226111 17123 9171 19113 7 1248 518116 2131465 12218 381200 115 52172 11216 225130 110118 4103 324118 48113 417 1 1241201 2261250 11319 114120 124114 33131

113120

201126 20101123 7 17

1231 518 54 500m3km 10km 327269627

Phylogenetic tree of H5 HA genes of the HPAIV isolates

A/chicken/Guiyang/3055/2005 2.3.3 A/goose/Guiyang/3422/2005 2.3.3 A/duck/Guiyang/3242/2005 2.3.3 A/duck/Hunan/139/2005 2.3.1 A/d k/H A/duck/Hunan/149/2005 /149/2005 2 2.3.1 31 A/duck/Hunan/127/2005 2.3.1 A/chicken/Shantou/810/05 A/duck/Guangxi/89/2006 A/duck/Yunnan/1126/2006 A/goose/Guangxi/345/2005 A/d k/H A/duck/Hunan/1265/2005 /1265/2005 A/duck/Vietnam/568/2005 A/goose/Guangxi/3316/2005 A/feral pigeon/Hong Kong/3409/2009 A/oriental magpie robin/Hong Kong/9298/2009 A/grey heron/Hong Kong/779/2009 A/ A/grey h heron/Hong /H K Kong/1046/2008 /1046/2008 A/crested myna/Hong Kong/1178/2009 large billed crow/Hong Kong/885/2009 A/great egret/Hong Kong/807/2008

2.3.3

231 2.3.1

234 2.3.4

A/whooper swan/Aomori/1/2008 A/whooper swan/Aomori/2/2008 A/whooper / swan/Akita/1/2008 / / / A/whooper swan/Hokkaido/1/2008

A/chicken/Primorje/1/2008 A/chicken/Korea/Gimje/2008

A/whooper swan/Hokkaido/2/2008
A/chicken/Laos/LPQ001/2008 A/ hi k /L /LPQ001/2008 A/chicken/Hunan/8/2008 A/black-crowned night heron/Hong Kong/659/2008 A/Hong Kong/6841/2010 A/great crested grebe/Tyva/120/2009 A/great crested-grebe/Qinghai/1/2009 A/ h A/whooper swan/Mongolia/1/2010 /M li /1/2010 A/ grebe/Tyva/2/10 A/bar-headed goose/Mongolia/X25/2009 A/common goldeneye/Mongolia/X60/2009 A/whooper swan/Mongolia/2/2009 A/whooper swan/Mongolia/4/2009 0 01 0.01

2.3.2

A/duck/Hokkaido/WZ83/2010 / / / / A/chicken/Shimane/1/2010
A/whooper swan/Mongolia/21/2010

Ck/Mexico/232/94 Ck/Taiwan/1209/03 Dk/NY/185502/02 Emu/Texas/39442/93 Ty/MN/3689-1551/81 Mallard/Ohio/345/88 Mallard/Wisconsin/169/75 Ty/Wisconsin/68 Ck/Florida/22780-2/88 Ck/Pennsylvania/10210/86 Ck/Pennsylvania/1/83 Tern/SouthAfrica/61 Ck/Scotland/59 Ty/England/N28/73 Mallard/Miyagi/53/76 Dk/HongKong/205/77 Dk/Primorie/2633/01 Ck/France/03426a/03 Mallard/Sweden/2/02 Mallard/Netherlands/3/99 Ck/Italy/312/97 HongKong/156/97 Ck/HongKong/728/97 HongKong/483/97 Gs/Guangdong/1/96 Ck/Hebei/326/05 Ck/Fujian/1042/05 VietNam/1203/04 Ck/Thailand/73/04 Thailand/2-SP-33/04 Hanoi/03/04 Ck/Vietnam/NCVD09/05 Gs/Cambodia/022b/2005 Ck/Cambodia/022LC3b/05 HongKong/213/03 Ck/Yamaguchi/7/04 Ck/K t /3/04 Ck/Kyoto/3/04 Ck/Korea/ES/03 Mallard/Italy/332/06 Bh goose/Mongolia/1/05 Bh Goose/Qinghai/5/05

Ck/Ibaraki/1/05 Ck/Guatemala/45511-1/00

2005

2004

H5HA

0.02

Ck/Miyazaki/S749/07 Mountain hawk-eagle/Kumamoto/1/07 Ck/Okayama/T6/07 Ck/Miyazaki/H358/07 Ck/Miyazaki/K11/07 Dk/Indonesia/MS/04 Ck/Yunnan/447/05 Dk/Fujian/1734/05 Ck/Hunan/999/05 Ck/Guangxi/604/05 Dk/Vietnam/568/2005 Whooper swan/Hokkaido/2/08 Whooper swan/Hokkaido/1/08 Whooper swan/Akita/1/08

2007

2008

Gene derivation of the swineswine-origin influenza A (H1N1) virus

North Classical Cl i l swine i Human (derived from the American (H3N2) avian 1918 virus) Eurasian avian-like swine

PB2 - North American avian PB1 - Human H3N2 PA - North American avian H1 - Classical swine NP - Classical swine N1 - Eurasian avian avian-like swine M - Eurasian avian avian-like swine NS - Classical swine At least 18,366 deaths in 214 countries as of 18 July 2010

Each of the pandemic strains have been generated in pigs. Genetic reassortment often occurs in birds and pigs. The H1N1 strain is a genuine swine influenza virus.

Swineorigin InfluenzaA virus(H1N1)

Modified from Novel Swine-Origin Influenza A (H1N1) Virus investigation Team, N Eng J Med, 2009

goose

turkey 6-9Months quail Low Pathogenic AIV chickens

HPAIV

duck Human virus pig Apathogenic AIV

X
Genetic Reassortant virus

Pandemic virus

HPAI virus and human pandemic virus strains

Library of vaccine strain candidates

Influenza viruses of 65 combinations of th HA and the d NA subtypes have been isolated from fecal samples of ducks in Alaska, Siberia, Mongolia, Taiwan, China, and Japan (bl k) (black). 79 other combinations have been generated by genetic reassort-ment procedure in the lab ( d) (red)..

Isolates from water birds (65 combinations)

Reassortants g generated in the lab (79 combinations)

Thus, 246 avian influenza viruses of 144 combinations of HA and NA subtypes have been stocked as vaccine strain candidates. candidates Their pathogenicity pathogenicity, antigenicity, antigenicity genetic information and yield in chicken embryo have been analyzed, databased, and opened for Web site (http://virusdb.czc.hokudai.ac.jp/vdbportal/view/index.jsp).

Are we prepared for pandemic flu?

1. H1 to H16 and N1 to N9 subtypes of influenza A viruses perpetuate in the lakes where ducks nest in nature. 2 Vir 2. Viruses ses maintained b by nat natural ral reser reservoir oir are antigenically antigenicall and genetically stasis. 3. 1957 H2N2 and 1968 H3N2 viruses are reassortants between avian influenza virus and the preceding human strains. 4. Pigs are susceptible to both of avian and mammalian viruses and generate reassortants. 5. Avian viruses of any subtype can contribute genes for reassortants: N None of f th the 16 HA and d 9 NA subtypes bt can be b ruled l d out t as potential t ti l candidates for future pandemics. 6 Surveillance of swine flu is crucial especially in the countries where 6. avian flu has not been controlled. 7. Preparedness p for pandemic p flu should be based on the measures for the control of seasonal flu.

How should we control avian influenza ? Are we prepared for pandemic influenza ?
1. Why have the H5N1 HPAIVs persisted in poultry for 14 years and been antigenic variants selected ? Misuse of Vaccine. 2. Will the HPAIVs returned to migratory birds persist in nature ? Started perpetuation of HPAIVs in the nesting lakes of ducks. ducks 3. How should avian influenza be controlled in poultry ? Stamping-out Stamping out without misuse of vaccine is only way, so far. 4. What are the advantage and disadvantage of the use of vaccines ? 5 Is it OK to forget about H5N1 avian influenza virus strain after the 5. swine-origin H1N1 strain emerged ? 6. Are the measures for the control of seasonal flu satisfactory y? The measures how to control pandemic influenza should be based on the measures for the control of seasonal influenza.

Global surveillance and seasonal flu control measure-based strategy

How are we then prepared p p for emerging g g zoonoses ?


Y Year 1977 1982 1983 1986 1989 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 Di Disease Ebola hemorrhagic fever Hemorrhagic fever with renal syndrome

Emerging Zoonoses
C Causative ti agent t Ebola virus of Filoviridae Hantavirus of Bunyaviridae C t Country Zaire (DRC) Korea USA USA UK USA Venezuela USA USA Brazil Australia South American countries Z i Zaire Australia Hong Kong India Malaysia Thai, Philippines, Asia Saudi Arabia, Yemen USA, Canada, Mexico Congo Hong Kong, China, worldwide Netherland, Belgium, Germany China and 11 countries of Asia, Fareast, and Africa Angola Congo H Human cases 318 (279 daeths) 388 73,000(61 deaths) 9,000 , 113(-2001) 1,200 105(35 deaths) 22,000 80 (50 deaths) 26 (10 deaths) 2 (1 dath) 239 (30 deaths) 318 (249 deaths) d th ) 2 deaths 18 (6 deaths) 16 (4 deaths) 258 (100 deaths) >30,000 (>700 deaths) 884 (124 death) 17,000 (>600 deaths) 210175 deaths 8,437 (813 deaths) 86 ( death) 331204 deaths 424363 deaths 1210 deaths

Bloody diarrhea, Hemolytic uremic syndrome Escherichia coli O157:H7 Lyme y disease Borreria burgdorferi g HIV of Retroviridae AIDS BSE Prion Ehrlichiosis Ehrlichia chaffensis, E. phagocytophila Venezuelan hemorrhagic fever Guanarito virus of Arenaviridae Cat scratch disease Bartonella hensele Sin Nombre virus of Bunyaviridae Hantavirus pulmonary syndrome Brazilian hemorrhagic fever Sabia virus of Arenaviridae Hendra virus of Paramyxoviridae Morbillivirus infection from horses Hantavirus pulmonary syndrome Ebola Eb l hemorrhagic h h i fever f Rabies from bat Avian influenza virus (H5N1) infection Pneumonic plague Nipah virus infection Leptospirosis-present Rift Valley fever West Nile fever

Hantavirus of bunyaviridae Eb l virus Ebola i of f Filoviridae Fil i id Lyssavirus type 7 of Rhabdoviridae

Highly pathogenic avian influenza virus

Yersinia pestis
Morbillivirus of Paramyxoviridae

Leptospira interrogans
Rift Valley fever virus of Bunyaviridae West Nile virus of Flaviviridae Ebola virus of Filoviridae Coronavirus Highly pathogenic avian influenza virus Highly pathogenic avian influenza virus Marburg virus of Filoviridae Ebola virus of Filoviridae

20012007 2001 Ebola hemorrhagic g fever 2004 2003 SARS

Avian influenza virus (H7N7) infection 2004 Avian influenza virus (H5N1) infection 2008 2004 Marburg hemorrhagic fever 2005 2005 Ebola hemorrhagic fever

Emerging Zoonoses
Many of the agents responsible for epidemics throughout human history have their origins in animals. Nearly all emerging disease episodes of the past 40 years have involved zoonotic infectious agents. Episodes of zoonoses are increasing around the globe. Zoonoses should be controlled when the novel field of science for the control of zoonoses is established by the collaboration with and fusion of Veterinary y Medicine, Medicine, Public Health, Ecology, Epidemiology, Entomology, Pharmacology, Social Sciences and Computer Science.

The nature of the threat of new, emerging and re-emerging re emerging infections
Climate, weather, rainfall, temperature (global warming) Population movements and the intrusion of humans and domestic animals into arthropod p habitats Deforestation and settlement of new tropical forest/farm margins Expanding E di primitive i iti irrigation i i ti systems t The opening up of isolated ecosystems such as islands Increased long-distance air travel Increased long long-distance distance livestock transportation New routings of long-distance bird migrations Uncontrolled urbanization and environmental pollution

For the Control of Zoonoses

Veterinary Medicine
Ministry of Agriculture, Forestry and Fisheries Ministry of Education, Culture, Sports, Science and Technology

Medicine
Ministry of Health, Labour and Welfare

Transmission of Pathogens
Prevention and therapeutics of diseases in livestock Ecology of zoonotic pathogens Prevention and control of zoonoses Maintenance and improvement of human and public health

OIE

WHO

Ministry of the Environment Species conservation of wild animals

For the control of zoonoses

Identification of natural host & elucidation of the route of transmission

CONTROL OF ZOONOSES
Development of measures for diagnosis and prevention Clarification of the molecular basis of pathogenesis

Zoonoses should be controlled when the novel field of science is established by the collaboration with and fusion of Veterinary Medicine Medicine, Medicine Medicine, Public Health Health, Ecology, Epidemiology, and Computer Science.

Research and Development Model for Zoonosis Control

Panel of antisera Prediction of antigenic drift Prediction of pandemic influenza virus subtypes yp Mucosal vaccine

Vaccine strain candidates

Global surveillance
Isolate from ducks Genetic reassortant

Molecular basis of pathogenisity

Interspecies transmission

Antivirals

Department of Global Epidemiology Identification of natural host animals of zoonotic pathogens Genetic analysis of pathogens Database development of genome information Prevention and control of zoonoses

Department of Molecular Pathobiology Diagnosis of zoonotic diseases Identification of determinant for host specificity Molecular basis of pathogenicity Development of rapid and highly sensitive detection methods of zoonotic pathogens

Department of Bioresources P Preservation ti and d supply l of f zoonotic ti pathogens, cells, genes, antibodies and animal strains Development of prevention and prophylactic measures

Department of Collaboration and Education Coordination of collaboration programs with international and domestic organizations Training of experts for the control of zoonoses Improvement of IT infrastructure for the international collaboration for research and education

H kk id University Hokkaido U i it Research R h Center C t for f Zoonosis Z i Control C t l

Hokudai Center for Zoonosis Control in Zambia Identification of natural host animals and transmission routes of zoonotic pathogens in Africa Comprehensive screening of unknown pathogens in Africa Prevention and control of zoonoses

Global Surveillance International Collaboration

World Health Organization (WHO) World Organization for Animal Health (OIE) Food and Agriculture Organization (FAO) Centers for Disease Control and Prevention (CDC)

Research Center for Zoonosis Control

CZC Facilities
BSL2 and BSL3 animal room Bioclean room Ultramicromorphology room P2 and P3 room Cold room Warm room W Freezer room Meeting room Office room

2nd floor

1st floor

BSL3 Area

BSL2 Area

Bioclean Area

Office Area

Engineering

Others

BSL-4 containment in Canada

Isolate from ducks Genetic reassortant

Library of influenza virus strains

Development of vaccines

Development of antivirals

Surveillance of avian influenza in Asia

Surveillance of hemorrhagic fever virus in Africa

Surveillance of Trypanosoma in Africa

Surveillance of mycobacteria in Asia

Pathogenesis

Structural analysis of virus proteins

Training course in Japan

Training course in Sri Lanka