OIE Collaborating CenterMember of OIE SCAD

Mechanism of FMD outbreaks and the Control of FMD in East Asia

Exotic Disease Research DivisionNational Institute of Animal Health

6-20-1 Kodaira, Tokyo 187-0022, Japan

Kenichi Sakamoto

Symposium on Prevention and Control of Foot and Mouth Disease

December 1, 2011

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General Information of Asia

1. Human PopulationWorld population 7.0 billions (2011.11)More than 60% 4.2 billions in Asia

2. Populations of domestic animals(2007 FAO)

World Population Ratio of AsiaCattle 1.38 billion 33.3%

Pigs 0.92 billion 58.5%

Sheep 1.09 billion 35.5%

3. Rapid Increase of economic activities in Asia RegionActive movements of people, animals, animal commodities

billion

Red line: Total populationBar: increasing population year by year

POOL １O,A,Asia1

POOL 7O&A

POOL 5O,A,SAT1,2

POOL ３O,A,Asia1

POOL ２O,A,Asia1

POOL 6O,A,SAT1,2,3

POOL４O,A,SAT1,2,3

Model of related FMDVs Distribution

（OIE/FAO_WRLFMD､；OIE/FAO Global Conf. on FMD, Paraguay, 2009）

3 pools covering Europe, Middle-East and Asia3 pools covering Africa1 pool for the Americas

FMD virus in Asia

POOL １O,A,Asia1

POOL ２O,A,Asia1

Asia1(G-V)

Asia1(G-IV)

AASIA

AASIA

OCathay

OSEA

OME-SA

(PanAsia-2)OME-SA

(PanAsia)Asia1(G-III)

（ Hammond et al. http://web.oie.int/eng/A_FMD2009/FMD_presentation/Session%202_1/2_1_1_Hammond.pdf）

The recent characteristic of FMD outbreaks in the region

• The FMD spreads more quickly than before.

• FMD outbreaks are predominantly caused by FMDV serotype O.

• The two main topotypes involved are South-East Asia (SEA) and Middle East – South Asia (ME-SA).

• FMDV of the SEA topotype (Mya-98 lineage) is widespread in South-East Asia and East Asia.

• The economic impact of FMD in East Asia (People’s Republic of China, Japan and Korea) has been severe in 2010–2011.

• FMD outbreaks due to serotype A have been sporadically observed in recent years.

• Serotype Asia 1 newly appeared in Pakistan from 2010 and Bahrain and Iran in 2011.

FMD situation in the Asia

Distribution of Serotype O in the Asia

FMD situation in the region (2009-2011)Type O

SEA topotype (Mya-98 lineage), Currently widely circulating in Asia. Lao PDR (2008-2009), Myanmar (2008-2009) , Thailand (2009), Malaysia (2009), Hong Kong SAR (2010), China (2010). In East Asia the Republic of Korea (South Korea)(2010.4, 2010.11-2011.4), Japan (2010.4), Russia (2010. 7), Mongolia (2010. 8), The Democratic People‘s Republic of Korea (North Korea) (2010.12- 2011).

CATHAY topotypeVietnam (2008), Taiwan (2009-2011) .

ME-SA topotypeBangladesh (2009), Malaysia (2009), (PanAsia lineage). Bhutan, Bangladesh and Nepal (2010), (the O-Ind-2001 lineage). China (2011.3) (PanAsia lineage closely related to Vietnam isolates).Afghanistan, Iran, Pakistan and Turkey (O-PanAsia-2)

Unknown topotypeKazakhstan (2011.5)

9

Distribution of Serotype A in the Asia

FMD situation in the region (2009-2011)Type A, Asia 1, C

Type AFMD type A (ASIA topotype): wide circulation in the region. Lao PDR (2008), Thailand (2009), Vietnam (2008-2009), P.R. China (2009), the Republic of Korea (2010.1-3).

Myanmar (2010. 9), very close to the border with Bangladesh.By the phylogenetic analysis the virus was most closely related to viruses occurring in India in 2000.

Asia 1The recent appearance of new Asia 1, Pakistan (2010) Bahrain and Iran (2011).

C No outbreak in the region in 2009-2011

National Institute of Animal Health (Exotic Research Station)

BSL 3FMD diagnosisFMD research works

ＲＴ－ＰＣＲ

Virus Isolation

negative

positive

Antigen detection ELISA

Isolatedvirus

Neg

Cont.OACSAT1SAT2SAT3Asia1SVD

Strong Weak

Cont.

ＲＴ－ＰＣＲ

Antigen ＥＬＩＳＡ

Virus isolation

2.5 ～ 6 hour

4 ～ 6 hour

2～7days

Consuming time for diagnosis

FMD Diagnosis Methods in Japan

Antibody Detection(in outbreaks and surveillances)•LPB ELISA•NT (in necessary)

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292 outbreaks(including 1 goat

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牛発生農場数 豚発生農場数

（戸） （戸）

発生事例数

No. of Outbreaks per day

Number of outbreaks (Cattle and Swine)

Total No. of Outbreaks

No. of Outbreaks in cattle

No. of Outbreaks in swine

Date

More wider spread

Large number of animals were waiting to be slaughtered

Japan decided FMD vaccination

May 19Restriction Area

Start vaccination to all the animals in movement control region (in Red circles) to control the speed of FMD

May 22Restriction Area

- 181 cases

40km

First time Vaccination in Japan

• All cloven-hoofed animals in the movement restriction zone (10 km radius)

• 6 PD50 O Manisa (oil adjuvant killed vaccine)

• Estimated r1 value 0.7

• Purified inactivated

• Destruction of the vaccinated animals

Cattle 45,950

Pigs 79,606

Others 118

Total 125,556

Number of vaccinated animals

1,066 farms

Vaccine matching of FMD in the region

• O SEA topotype (Mya-98): O Manisa, O Ind R2/75 and O Taw 98(some isolates from Hong Kong (2011) : not matching O Manisa, O Taw 98)

• ME-SA topotype (PanAsia-2) (2009-2010) : O Manisa, O IND R2/75, O TAW 98 and O BFS.

• FMDV A from Iran (2009-2010) : A TUR 06.

• FMDV A from Afghanistan (2010-2011): A IRAN 05, A TUR 06.

• A from the People’s Republic of China and the Republic of Korea: Mya-97.

• Asia 1 from Bahrain, Pakistan and Iran (2010-2011):

not matching with Asia 1 IND 8/79, Asia 1 Shamir, Asia 1 WBN

17

Foot-and-Mouth Disease Virus

7 immunologically distinct serotypes

Capsid Non-structural proteins

AAAAAAAAAn

L P1 P2 P35’ UTR 3’ UTRVPg

VP4 VP2 VP3 VP12A 2B 2C 3A 3B 3C 3DPoly C

~8.2 kb

IRES

O, A, C, Asia 1, SAT 1, SAT 2, SAT 3

FMD Differentiation of Infection and Vaccination

FMD VaccineInactivated Vaccine (concentrated purified)Potency：Normal ３ＰＤ５０

For emergency ６～１２ＰＤ５０

Differentiated Infected VaccinatedAnimal

１）LPB ELISA ○ ○

２）NSP ELISA ○ ×

The conditions for FMD vaccine

•The FMD vaccine must be an inactivated vaccine.

•The vaccine selected should be a good antigenic match for field isolates. It is very important to send samples to FMD Reference Laboratories for virus isolation and sequencing.

•Whenever FMD vaccine is used, in an emergency to control FMD outbreaks or routinely, it is essential to be able to determine whether FMDV antibodies are the result of infection or vaccination.

•The vaccine should be produced in accordance with the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals.

•Cooperation and research collaboration on FMD diagnosis and vaccine production are important between Member Countries to control FMD in the region.

•In order to be able to differentiate between antibodies by infection and by vaccination, there is an urgent need to continue striving to produce pure vaccines to limit the effect of NSP’s when evaluating surveillance results.

Empty Capsid

Intact FMDV

Empty Capsid of FMDV as vaccine antigen

Virus Genome

One Copy of 3D Protein

No Genome

No NS proteins

Only Outer Shell

Genetic engineering technique

A new generation FMD vaccine

1. Sharing Disease Information in Pool 1 - 3(South East and East, Central and Middle-East Asia)

2. Early notification of the FMD to Members in the region and OIE

3. Strengthen the border control to prevent FMD virus entry 4. Scientific research collaborations of the FMD laboratories

to control the disease among the member countries in the region.

5. Technical supports to the developing countries in the region for diagnosis of FMD

6. Financial and economic supports to provide good matching FMD vaccines to FMD epidemic countries

To reduce FMD outbreaks in the regionProposals

The epidemiological roles of susceptible animals in FMD

Cattle：The most susceptible livestock to FMDV.

（Detector）

Pigs：Pigs excrete very large quantities of the virus (100- to 2000-fold more than cattle and sheep).

（Amplifier）

Sheep：Mild or un-apparent clinical signs, making early detection of FMDV infection difficult.

（Transporter ?）

An example of Detector

• Japan in 2000: PanAsia of O ME-SA topotype• FMD in only cattle

• Miyazaki, Hokkaido killed 7４０ cattle• Only Japanese black cattle created atypical

clinical signs. No vesicles in mouth or nasal cavity

• Pigs form typical symptom of vesicle in foot by animal experiments

• The source of the virus was considered to be imported hay or straw from the Asia region.

Excretion of virus

Minimal effectiveinfecting dose 101.0 ID50 102.6 ID50

105 ID50 108 ID50

Detector Amplifier

Epidemiology of FMD

Epidemiological role

Examples of Amplifier

• Taiwan 1997, UK 2001, South Korea 2002

• Recently Japan 2010, South Korea 2010-2011

• O type Cathay, ME-SA and SEA topotypes

• Accumulation of FMDV in the environments makes new outbreaks.

• FMD outbreaks in pigs are often on a very large scale.

• The FMD can cause serious economic damage.

Economic Impact in Recent FMD Outbreaks

Year Country No. of Animal Destroy Economic Damage

1997 Taipei China Swine 4 million 3.6 Billion US$ in first year

2000 Japan Bovine 740 72.7 Million US$

2000 S. Korea Bovine 2200 273 Million US$(FMD Vaccine to 850,000 Bovine)

2002 S. Korea Swine 160,000 225 Million US$

2001 UK Susceptible Animals 14.4 Billion US$6 million

2010 Japan Swine & Bovine 290,000 3 Billion US$ for complete recovery

2010- S. Korea Swine & Bovine 3.5 Million 3 Billion US$

Samples of Transporter

• UK in 2001, Cyprus in 2007

• The clinical signs are sometimes mild or unapparent.

• It makes early detection of FMDV infection difficult.

• Possibility of huge FMD outbreaks

• Serological tests are important for the diagnosis of FMD in sheep.

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発生農場数

英国の口蹄疫発生(MAFF UK, 2001～2002)

累積発生数

１日の発生数Numbers of slaughtered animals

2002. 1.14

Cases：2026

slaughtered 4.05 Million

species cattle 0.6 M

sheep 3.3 M

pig 0.14 M

goat 2 Thousand

others 2 T

由来農場Freshwater / Isle of Wight

発見されたと畜場（27頭）Brentwood / Essex

2001.2.21.

由来農場Great Horwood / Buckinghamshire

由来農場Freshwater / Isle of Wight

発生と畜場･農場（４件）Essex州

UK FMD outbreaks in 2001

2001.2.25

e

発生農場（３件）Devon州

発生農場（３件）Northumberland州

移動制限地域Berwick-on-Tweed / Northumberland州

移動制限地域Fyvie / Abardeen州

移動制限地域Woodchester 及びKington

/ Gloucestershire州

発生農場（１件）Hereford州

Epidemiological findings of 2001 UK FMD outbreaks

19 heads of sheep

Livestock markets

Distance between A B 400km

C happened before A and B

A Initial outbreak （Tyne and Wear state）

Pig Farm

Origin of the FMDV

Unknown（Airborn infection）

C Cumbria stateDevon state

Expansion of Nationwide infection

B First find Essex state Slaughter house

（Airborn infection）

Cattle near by the slaughter house

（Epiological Investigation）

To prevent pandemic outbreaks and to inhibit huge economic catastrophes in the region

1. Early Detection

2. Early Extinguishment

3. Early notification

4. Good matching Vaccine

However….

Infected pigs excrete FMD viruses within 2 - 4 days after FMD infection

And it takes about 7 days before the animals can induce protective antibodies by vaccination.

Prompt effective tools are strongly desired

◎ Anti-FMDV Agents in vitroGroup of Compound Agent IC50(μg/ml)

Anti-HIV agents (Non-Nucleotide Efavirenz 20 - 40Reverse Transcriptase Inhibitors)

Pyrazinecarboxamide derivatives T-705 14T-1105 1.6T-1106 17

Virus : FMDV O/JPN/2000 Cells : IBRS-2

FMDV Serotype Challenge dose EC50(μg/ml)

A 22 Iraq 10TCID50 0.44

C Philippine 10TCID50 1.76

Asia 1 Shamir 101.5TCID50 0.88

◎ Anti-FMDV Activities of T-1105 to FMDV serotypes in vitro

Mechanism of Action :

Inhibition of viral RNA polymerase

Plas

ma

conc

. (µg

/mL

)

Pig Cattle Sheep

Pharmacokinetics of T-1105 in domestic animals

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100

0 10 20 30hour

T-1105T-1105M1

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T-1105T-1105M1

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T-1105T-1105M1

T-1105 Oral single Administration

100mg/kg Body Weight

Concentration of T-1105 in pig is high and poor in cattle

T-1105 (Twice a day, 200mg/kg)

1 dpi 2 dpi 3dpi 4dpi 6dpi 8dpi0 dpi 5dpi 7dpi

Collection of plasma, serum and nasal swabs

Virus inoculation

Experimental Schedule

Initial Administration of T-1105 is just one hour before FMDV inoculation

Oral administration (blended in feed)

Virus: O/JPN/2000, 106 TCID50/ml

Site of injection: Left front heel bulb intradermal

T-1105 Administrated Group・ No Clinical signs

・ No virus Excretion

Control group・ Typical Clinical signs

Clinical Signs after the virus inoculation

＋Creation of vesicular legion++ Rupture of vesicle Lameness and difficulty in standing

0 1 2 3 4 5 6 7 81 － － － － － － － － －2 － － － － － － － － －3 － － － － － － － － －4 － － － － － － － － －

5 － － ＋ ++ ++ ++ ++ ++ ++6 － － － ＋ ＋ ++ ++ ++ ++

T-1105

Control

Treatment No.Days post inoculation

In vivo test with virulent FMDV O/TAW/1997Assay of the pathogenicity of the virus by * PHID50

*50 % Pig Heel infectious Dose : The creation of vesicula in the injection site within 48 hours

Virus PHID50 (pfu) Ratio of virulence

O/JPN/2000 2.5 x 105 1

O/TAW/1997 9.4 x 101 2.5 x 103

O/SKR/2000 2.2 x 103 102

O/UKG/2001 1.9 x 103 10 2

“ O/JPN/2000 is a low virulent FMD virus to pigs”

Introduction of O/TAW/1997 for the in vivo efficacy test of T-1105 in pigs

1 ２ ３ ４ ５ ６ ７ ８ ９

1 ＋ ＋ ++ ++ ++ ++ ++ ++ ++6 ＋ ＋ ++ ++ ++ ++ ++ ++ ++

T-1105Admin.

Non-admin.

＋ Creation of vesicular legion++ Rupture of vesicle

Pig No.Days post infection

2 － + + + ++ ++ － － －3 － － － － － － － － －4 － － － － － － － － －5 － + + + ++ ++ － － －

Lameness / Difficulty in standing

Summary of clinical signs in O/Taiwan/97 infection

No Lameness / No Difficulty in standing

Treated: Clinical signs (5dpi)

#2

T-1105 admin. pig

Control Clinical signs (5dpi)

#1 #6

Non-admin. pig

#2 #3

#5 #4

Treated: Clinical signs (7dpi)

#1 #6

Control: Clinical signs (7dpi)

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102

103

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106

0dpi 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi 7dpidays post infection

PFU

/0.1

ml

#1

#6

Non-admin. group

Virus excretion in saliva from pigs infected with O/Taw/97

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0dpi 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi 7dpidays post infection

PFU

/0.1

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T-1105 admin. group

#2 #3#4#5

Clinical signs in pigs infectedwith O/Taiwan/97

Two of admin. pigs made vesicle at the only inoculation site.

The low level of virus was detected from saliva of some pigs.

Brief and lower level of viremia were detected in some pigs.

One pig did not show viremia as well as clinical signs.

FMDV excretion and viremia were controlled by administration of the T-1105

Minimum Effective dose of T-1105On Day 7

Group Concentration of T-1105(mg/kg)

No. of pigsTested / Protected

Protection(%)

A 3.125 6/5 84

B 12.5 6/3 50

C 50 6/6 100

D 200 6/6 100

E (Control) 0 6/1 16

Minimum effective doses of T-1105 conclusion

・ Day 7: Group C (50mg) Protection 100%

Group D (200mg) 100%

・ Day 10 : Group C Protection 84%

Group D 100%

・ 50 - 100mg/kg is the minimum dose to control

FMD in pigs

Virus : FMDV O IND R2/75

Conclusion

T-1105 administrated pigs with FMDV challenge・ No Viremia, low level fo viremia with highly virulent strain

・ No Clinical Signs (No Vesicles or only injection site, No Lameness)

・ No virus Excretion or very low level

Advantage of anti-FMDV compound, T-1105・Prompt effect ( In one hour)

・Independence from FMDV serotypes or strains

・ Administrating with food

Model for utilization of T-1105

FMD outbreaks in pig* Early stage of the outbreak

Objective: Reduction of FMD spread

Result: Decreasing the FMDV in the zoon

and infection in pig farms and the cattle farms

A

B

C

D

E

Zoon

F

3 km

Administration of the antiviral agent T-1105 to all pigs inPig farms within 3 km (Farm A, B, C, D, E, F)

Recommendation to use the antiviral agent

T-1105 should be used in the following conditions

1. In FMD outbreaks in pigs

2. Emergency use

3. To the normal pigs surrounding of an infected pig farm

4. Administrated pigs should be slaughtered to reduce the risk of emerging a mutant virus

5. T-1105 is administrated with food orally6. Administrated pigs should not be used for human consumption

until T-1105 is proved to be safe for human health

7. Strictly regulated by the countries and regions like as the emergency FMD vaccine

POOL １O,A,Asia1

POOL 7O&A

POOL 5O,A,SAT1,2

POOL ３O,A,Asia1

POOL ２O,A,Asia1

POOL 6O,A,SAT1,2,3

POOL４O,A,SAT1,2,3

Model of related FMDVs Distribution

（OIE/FAO_WRLFMD､；OIE/FAO Global Conf. on FMD, Paraguay, 2009）

3 pools covering Europe, Middle-East and Asia3 pools covering Africa1 pool for the Americas

FMD virus in Asia

POOL １O,A,Asia1

POOL ２O,A,Asia1

Asia1(G-V)

Asia1(G-IV)

AASIA

AASIA

OCathay

OSEA

OME-SA

(PanAsia-2)OME-SA

(PanAsia)Asia1(G-III)

（ Hammond et al. http://web.oie.int/eng/A_FMD2009/FMD_presentation/Session%202_1/2_1_1_Hammond.pdf）

Cattle Movement

Cattle (millions)

Buffalo(millions)

Import Export Comparative Unit Price Ranking

Cambodia 3.34 0.72

- +++ **Moderate

LowLao PDR 1.35 1.16

- +++ **Moderate

LowMalaysia 0.8 0.14

+++ - ***** HighMyanmar 12.63 2.84

- ++++ * LowThailand 9.34 1.58

++ ++ *** Moderate Vietnam 6.88 3

+++ - ****Moderate

High

Cattle/Buffalo Trade vs. population and price

Recent FMD outbreaks in East Asia

1)1997- O: Cathay topotype: Taiwan (Nationwide) Illegal Movement of pigs from China

2) 1999-2002 O: ME-SA topotype, PanAsia lineage: China, Taiwan, Japan, Korea, Amur region of Russia, Mongolia

3) 2005- Asia 1:Hong Kong, China, Amur region of Russia, Mongolia, North Korea

4) 2009- A: ASIA topotype, China, South Korea

5) 2010-2011 O,SEA topotype, Mya-98 lineage: China, South Korea, Japan, Amur region of Russia, Mongolia, North Korea

China has the important role of the FMD outbreaks in East Asia1) Long border with FMD epidemic countries in South East and Central Asia2) Large numbers of susceptible animals (pigs, sheep, cattle)3) Rapid increase of economic activities (A raising nation)4) Active cross border movement of people and animal commodities

FMDV

The Scheme of FMD virus spread in East Asia

Mechanism of FMD spread in East Asia

FMD (Epidemic countries) China East Asia countries

199019911992199319941995199619971998199920002001

FMD – 10 year evolution of Asia topotype O

FAO warns same may happen again near future

Recent serotype O Outbreaks

Topotype SEA (Mya-98)

1. Sharing Disease Information in Asian Region(South East, Central and East Asia)

2. Early notification of the FMD to Members in the region and OIE

3. Strengthen the border control to prevent FMD virus entry 4. Scientific research collaborations of the FMD laboratories

to control the disease among China, Japan, Korea and Taiwan

5. Technical support to South East Asian countries for diagnosis of FMD

6. Financial and economic supports to provide good matching FMD vaccines to SEA countries

To reduce FMD outbreaks in the regionProposals

Thank you for your attention !!