swine influenza surveillance among pigs in nan province · 1918-19: spanish flu (h1n1) ... human...

Swine influenza surveillance among pigs in Nan Province

National Institute of Animal Health

Department of Livestock Development

Nan province

30 Aug 2011NIAH Thailand

การเฝ้าระวงัโรคไข้หวดัใหญ่สุกรในสุกรจังหวดัน่าน

สุจริา ปาจริยานนท์ สพ.บ. Ph.D.

สถาบันสุขภาพสัตว์แห่งชาติกรมปศุสัตว์จังหวดั น่าน

30 สิงหาคม 2554NIAH Thailand

Outline

• Introduction

• Materials & Methods

• Results

• Gaps and needs

• Acknowledgements

NIAH Thailand

Swine Influenza

First isolated from pigs by Shope in 1930

Worldwide distribution

High prevalence in North American and

European swine farms

All pathogenic SIV belong to type A

influenza viruses

NIAH Thailand

Influenza virus

Hemagglutinin (HA, H: 1-16)

Neuraminidase (NA, N: 1-9)

Wild waterfowl serve as global reservoir because they can be infected with virus of all 16 HA and 9 NA subtypes

SIV subtype defined by these two glycoproteins, the common subtype: H1N1, H3N2 and H1N2 other subtypes: H1N7, H3N1, H4N6 and H9N2

NIAH Thailand

Influenza virus

Enveloped RNA virus belongs to the family Orthomyxoviridae

8 RNA segmented genomes, genetic exchange and reassortment can generate during mixed infections

Infectious to human “zoonotic agent”

Pigs as “mixing vessel hosts” human and avian flu can undergo genetic reassortment and cause new pandemic human influenza viruses

Swine influenza virus

NIAH Thailand


Virus attaches to

respiratory tract and

causes destruction of

the airways of the

lung, resulting in

viral pneumonia

Influenza virus-induced

pneumonia in pig

NIAH Thailand

EM : virus particles in infected MDCK cell line

Virus particles

Deadly virus

1918-19: Spanish flu (H1N1)

• >500,000 dead in US, 20-50 mil.WW

1957-58: Asian flu (H2N2)

• 70,000 dead in US China->US

1968-69: Hong Kong flu (H3N2)

• 34,000 dead in US

1997: Hong Kong flu (H5N1)



2003: Avian flu (H7N7) Netherlands


2004-05 Vietnam, Thailand, Cambodia (H5N1)

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โรคไข้หวดัใหญ่เมก็ซิโก H1N1 2009

Confirmed death and infections

Unconfirmed infections

Confirmed infections

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Clinical signs of SIV

High fever (> 104OF)

Clear nasal discharge

Go off feed

Become inactive, huddle, pile together

Cough and/or sneeze

Have difficulty breathing

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Clinical signs of SIV

Usually last 5-10 days unless complicated

by secondary infections

• Pasteurella multocida

• Haemophilus parasuis

• Streptococcus pleuropneumoniae

• Mycoplasma hyopneumoniae

NIAH Thailand


Primary pathogen causes respiratory

disease or involves in PRDC

The most common isolates involve in PRDC

• PRRS

• Mycoplasma hyopneumoniae

• SIV

SIV causes an economic impact on pig

production, adds 2 more weeks for fattening

pigs to reach the market weight

(Esterday, 2002)NIAH Thailand

Swine influenza in Thailand

Only few reports regarding to SIV infection in

Thailand

Kupradinan et al. (1991), SIVs were isolated

from pigs with influenza-like illness in 1988,

the isolates were SIV H1N1

NIAH Thailand

A/Sw/Korea/CY08/07(H1N2)A/Sw/Korea/PZ4/06(H1N2)

A/Sw/Korea/Hongsong2/04(H1N2)A/Sw/Korea/S11/05(H1N2)

A/Sw/MN/16419/01(H1N2)A/Sw/OH/511445/07(H1N1)

A/Sw/Ohio/891/01(H1N2)A/Sw/MN/23124-S/01(H1N2)A/Sw/Illinois/100085A/01 (H1N2))

A/Sw/Indiana/9K035/99 (H1N2)A/Sw/Indiana/P12439/00 (H1N2)A/Turkey/MO/24093/99(H1N2)

A/Sw/Guangxi/13/06(H1N2)A/California/04/09(H1N1)A/Sw/North Carolina/93523/01 (H1N2)

A/Sw/North Carolina/98225/01(H1N2)A/Sw/Minnesota/55551/00 (H1N2)

A/duck/NC/91347/01(H1N2)A/Sw/CO/17871/01(H1N2)

A/Sw/Korea/CAS08/05(H1N1)A/Sw/Korea/CAN01/04(H1N1)

A/Sw/Iowa/930/01(H1N2)A/Sw/MO/1877/01(H1N2)

A/Sw/Korea/CY02/02(H1N2)A/Sw/MN/34893/01(H1N2)

A/Sw/Wisconsin/464/98(H1N1)A/Sw/Wisconsin/136/97(H1N1)

A/Sw/Wisconsin/238/97(H1N1)A/Sw/Zhejiang/1/04(H1N2)A/Sw/Shanghai/1/07(H1N2)

A/Sw/Guangdong/2/01(H1N1)A/Sw/Shanghai/1/05(H1N1)

A/Sw/MN/17138/01(H1N2)A/Sw/Wisconsin/457/98(H1N1)A/Sw/Wisconsin/163/97(H1N1)

A/Philippines/344/04(H1N2)A/Sw/Indiana/1726/88(H1N1)

A/Sw/Ratchaburi/NIAH1481/00(H1N1)A/Sw/Ratchaburi/NIAH550/03(H1N1)A/Sw/Alberta/56626/03(H1N1)

A/Sw/Ontario/57561/03(H1N1)A/Sw/Taiwan/CO935/04(H1N2)

A/Sw/Nebraska/1/92(H1N1)A/Sw/Ontario/11112/04(H1N1)

A/Sw/St-Hyacinthe/106/91(H1N1)A/Sw/Ontario/23866/04(H1N1)

A/Sw/Ontario/53518/03(H1N1)A/Sw/Saraburi/NIAH13021/05(H1N2)A/Sw/Chachoengsao/NIAH587/05(H1N1)A/Sw/Chonburi/NIAH589/05(H1N1)

A/Thailand/271/05(H1N1)A/Sw/Chonburi/NIAH9469/04(H1N1)

A/Sw/Chonburi/NIAH977/04 (H1N1)A/Sw/Miyazaki/1/06 (H1N2)A/Sw/Miyagi/5/2003(H1N2)

A/Sw/Saitama/96 (H1N2)A/Sw/England/283902/93 (H1N1)

A/Sw/England/117316/86A/Sw/Ehime/1/80 (H1N2)A/Sw/Hokkaido/2/81 (H1N1)A/Sw/Kentucky/1/76(H1N1)

A/Sw/New Jersey/11/76(H1N1)A/Sw/Tennessee/25/77(H1N1)

A/Sw/Minnesota/24/75(H1N1)A/Sw/Kyoto/3/79 (H1N1)A/Sw/Niigata/1/77 (H1N1)A/Sw/Iowa/4/76(H1N1)

A/Sw/Wisconsin/2/66(H1N1)A/Sw/Wisconsin/1/57(H1N1)

A/Sw/Wisconsin/1/61(H1N1)A/Sw/Illinois/63(H1N1)

100

1009898

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99100

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0.02

H1 Phylogenetic tree

Classical Swine lineage

Human lineage

Avianlineage

California isolateHuman case of classical swine H1Thai isolatesJapan isolates

NIAH Thailand

N1 Phylogenetic tree

A/Sw/Cotes dArmor/1488/99 (H1N1)A/Sw/Ille et Vilaine/1455/99 (H1N1)A/Sw/Cotes dArmor/1121/00 (H1N1)

A/Sw/Spain/50047/03(H1N1)A/Sw/England/195852/92 (H1N1)

A/Sw/Belgium/74/85(H1N1))A/Sw/Italy/1492/97 (H1N1)A/Sw/Italy/1513-8/98 (H1N1)

A/Sw/Italy/1509-6/97 (H1N1)A/Sw/Italy/1424-4/95 (H1N1)

A/Sw/Ratchaburi/NIAH550/03(H1N1)A/Sw/Ratchaburi/NIAH1481/00 (H1N1)

A/Sw/Chachoengsao/NIAH587/05 (H1N1)A/Sw/Chonburi/NIAH589/05 (H1N1)

A/Thailand/271/05(H1N1)A/Sw/Chonburi/NIAH9469/04 (H1 N1)

A/Sw/Chonburi/NIAH977/04 (H1N1)A/California/04/09 (H1N1)

A/Sw/Spain/51915/03(H1N1)A/Sw/Spain/53207/04(H1N1)

A/Sw/Cotes dArmor/1482/99 (H1N1)A/ Sw/Cotes dArmor/1515/99 (H1N1)

A/Sw/Virginia/670/87(H1N1)A/Sw/Virginia/671/87(H1N1)

A/Sw/Lot/2979/82(H1N1)A/Sw/Belgium/1/83(H1N1)

A/Sw/Finistere/3616/84(H1N1)A/turkey/France/87075/87(H1N1)

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0.02

H5N1 viruses

Classical Swine lineage

Human lineage

Avian lineage

Eurasian swine lineageCalifornia isolateHuman case of avian like swine N1Thai isolates

NIAH Thailand

Outline

• Introduction


• Results

• Gaps and needs


NIAH Thailand

Collection of samples

Nov 2010 - Feb 2011

Nasal swabs 880

Serum 880

Farm size: small farm < 30

medium farm ≥ 30-300

large farm > 300

NIAH Thailand

Nasal swab collection

Collect samples from early sick period especilly during febrile of illness

Samples should be collected on polyester, not cotton swabs

Swabbing the nasal passage for mucus obtained

Suspend the swab in a cold transport medium and keep cold

If the samples can not be process within 24 hours after collection, the samples should be storage at -70OC

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Transport media

Keep at 4OC: Medium + 0.5% BSA

For longer time keeping, -20OC is

recommended

Keep cold during tranportation

Inform the responsible person in the

lab before sending the samples

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Diagnosis

Virus isolation and identification

• Embryonated egg inoculation

• Tissue cultures: MDCK

PCR & RRT-PCR (typing/subtyping)

IHC for formalin-fixed paraffin-embedded tissue

Specimens: nasal/tracheal/bronchial swab, lung grossly visible bronchi and bronchiole

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Tissue culture

Eggs inoculation

(Two passages if no HA activity in the first passage)

Virus isolation

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Tissue culture: MDCK

RT-PCR

RT-PCR : detection and subtyping

of the influenza virus

PAM NS

NP

M H1 H3 N1 N2 M M H1 H3 N1 N2 M

M = Marker

1500 bp

1000 bp

500 bp722 bp

1082 bp

722 bp

1150 bp

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H1N1 H3N2

Real time PCR

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Antibody detection

HI (subtype specific) may have

some cross-reaction

ELISA: commercially available

for SIV H1N1 & H3N2 and type

A antibodies detection

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Antibody detection: HI test

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Outline

• Introduction


• Results

• Gaps and needs


NIAH Thailand

NIAH Thailand

Percentage of SIV subtype seropositive by HI test

11.7

8.48.1

1.9

0.6

0

2

4

6

8

10

12

14

All subtypes H1N1 550/46 H1N1 2009 H3N2 59/47 H3N2 Nongwa

All subtypes

H1N1 550/46

H1N1 2009

H3N2 59/47

H3N2 Nongwa

subtypes

Per

cen

tag

e o

f S

IV s

ub

typ

ese

rop

osi

tiv

e

n = 880

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

00

0Per

cen

tag

e o

f S

I vir

us

det

ecti

on

VIPCR

RRT-PCR

VI

PCR

RRT-PCR

methods

NIAH Thailand

Percentage of SI virus detection

Percentage of SIV seropositive among

difference farm size

19.4

9.9

4.8

0

5

10

15

20

25

30

Large Medium Small

Large

Medium

Small

% s

ero

po

siti

ve

Farm size

NIAH Thailand

Percentage of SIV seropositive among ages

0

5

10

15

20

25

30

35

40

45

50

1-5

(18/545)

6-10

(3/49)

11-15

(20/66)

16-20

(9/42)

21-25

(21/96)

26-30

(4/18)

>30

(27/64)

3.3

6.1

30.3

21.4 21.9 22.2

42.2 1-5 (18/545)

6-10 (3/49)

11-15 (20/66)

16-20 (9/42)

21-25 (21/96)

26-30 (4/18)

>30 (27/64)

%

sero

posi

tive

months (+/total)

n = 880

NIAH Thailand

Comparison of seropositive percentage of

swine influenza between HI test and ELISA

NIAH Thailand

0

2

4

6

8

10

12

14

All subtypes H1N1 H3N2

12.7

10.8

2.3

6.6

4.4

0.1

HI test

ELISA

%

sero

posi

tive

Subtypes

Outline

• Introduction


• Results

• Gaps and needs


NIAH Thailand

Gaps and Needs

Limitation of understanding of

- Viral epidemiology

- Transmission pigs humans

- Vaccine strain selection

Surveillance is crucial

Lab network

Sharing informations

NIAH Thailand

Outline

• Introduction


• Results

• Gaps and needs


NIAH Thailand

Acknowledgements

Veterinarians and technicians from Nan Provincial

Livestock Health Office

Pig production companies

Farmer from Nan Province

Colleagues from Bureau of Epidemiology, MoPH

US CDC

Staff from NIAH

NIAH Thailand

Thank youfor your attention

NIAH Thailand

swine influenza surveillance among pigs in nan province · 1918-19: spanish flu (h1n1) ... human...

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