FACULTY OF VETERINARY MEDICINE BOGOR …zoonosis.ipb.ac.id/pdf_file/zoonosis centre 19 April 2012-wayan-1.pdf · FACULTY OF VETERINARY MEDICINE BOGOR AGRICULTURAL UNIVERSITY, BOGOR.

FACULTY OF VETERINARY MEDICINE

BOGOR AGRICULTURAL UNIVERSITY, BOGOR

Faculty of Veterinary MedicineBogor Agricultural University

The Possibility of Subclinical Manifestation of AI as Source of

Infection to Human

Prof. I Wayan Teguh WibawanDr. R.Susanti

Prof. Retno D SoejoedonoProf. Gusti Ngurah Kade Mahardika

Surachmi Setyaningsih, Ph.DEkowati Handayani, Ph.D

Dr. Sri Murtini

Source of informations

• Research activity of Avian Influenza (AI)

with Banten Province

• Research activity and collaboration with

FAO

• Disertation of Dr. R. Susanti

• Outbreak of AI among commercial chicken

and backyard chicken in the field

Closed-Direct Contact

• In the daily life in Indonesia we are very

close to poultry

• Backyard Farm

• Interaction between species, chicken,

ducks, cats, dogs and human

• Bad quality of environment

Our Potrait

AI vaccinated Broilers

Very virulent, pathogenic

Potential as Zoonotic

Caused by highly pathogenic AI (HPAI) subtype H5N1

Avian Influenza (AI)

AVIAN INFLUENZA

• Orthomyxovirus

• Pleiomorphic

• envelope

• 8 segmen RNA

• 11 proteins

The Year of 2003

High morbidity and mortality

Blue comb

Haemorhagic symptoms among visceral organs

Bleeding of ovary and kidney

“Kaki kerokan”

AI outbreak in 2003

After the outbreak in 2003 and the used monovalent vaccines (H5N1, H5N2 and H5N9)

Vaccination applied to “commercial” as well as “back-yard” chicken

Vaccination depressed the incidence of AI significantly

Vaccination is choosed as one of strategy to control disease

Lack of knowledge and skills

Lack of “cold-chain”

Lack of coverage of vaccination

Lack of awareness

Problems of vaccination in back-yard chicken

Subclinical manifestation of AI infection

Healthy birds as carrier of virus

Challenge to human?

Wet-market, pooling area: source of infection

No more AI vaccination in back yard chicken is applied

The Impacts

The Year of 2006

The Role of Ducks as “Reservoir”

Sekuensing fragmen HA1

Subtipe

H5N1

Primer Sekuen basa

HA01F

HA645R5’ ATGGAGAAAATAGTGCTTCTTCTTGC’3

5’ GGAAATATAGGTGGTTGGGTTTTG’3

HA548F

HA1215R

5’CCAACCRAGARGATCTTTTGG’3

5’AYRGCCTCAAACTGAGTGTTC ’3

H5-155F

H5-699R5’ACACATGCYCARGACATACT’3

5’CTYTGRTTYAGTGTTGATGT’3

Designed by Dr.drh.IGN Mahardika & Dr. R. Susanti

645bp

667bp

545bp

Analisis domain penting

Analisis filogenetik

MEGA 3,1

Kabupaten Kecamatan Jenis Hewan Jumlah

Itik Entok Angsa

Sukabumi Cibadak 33 3 1 37

Cicurug 18 5 12 35

Cidahu 22 10 7 39

Nagrak 35 4 0 39

Bojonggenteng 35 8 5 48

Jumlah 143 30 25 198

Bogor Cibinong 8 23 10 41

Ciseeng 27 7 6 40

Cileungsi 20 17 0 37

Darmaga 32 3 6 41

Klapanunggal 20 10 6 36

Parung 25 6 1 32

Leuwiliang 22 7 6 35

Jumlah 154 73 35 262

Total 297 103 60 460

Sample collection from water fowl around breeding-and

commercial poultry farm

Kabupaten Kecamatan Subtipe Virus AI

H5N1 H5Nx HxN1 HxNx

Sukabumi Cibadak 0 0 2 0

Cucurug 0 1 1 1

Cidahu 1 1 2 1

Nagrak 1 0 2 4

Bojonggenteng 2 0 3 2

JumlahPrevalensi

4 2,02%

21,01%

105,05%

8 4,04%

Bogor Cibinong 1 1 2 0

Ciseeng 3 0 0 0

Cileungsi 2 0 0 0

Darmaga 0 0 1 0

Klapanunggal 3 0 0 0

Parung 4 0 0 0

Leuwiliang 4 0 0 0

JumlahPrevalensi

176,49%

1 0,38%

3 1,15%

0 0%

Jumlah TotalPrevalensi

21 4,57%

30,65%

132,83%

81,74%

P

R

E

V

A

L

E

N

c

y

No Isolat Asal

1 A/muscovy duck/Klapanunggal/IPB1-RS/2006 (H5N1) Klapanunggal

2 A/goose/Bojonggenteng/IPB2-RS/2006 (H5N1) Bojonggenteng

3 A/duck/Leuwiliang/IPB3-RS/2006 (H5N1) Leuwiliang

4 A/goose/Leuwiliang/IPB4-RS/2006 (H5N1) Leuwiliang

5 A/muscovy duck/Cileungsi/IPB5-RS/2006 (H5N1) Cileungsi

6 A/duck/Nagrak/IPB6-RS/2006 (H5N1) Nagrak

7 A/goose/Klapanunggal/IPB7-RS/2006 (H5N1) Klapanunggal

8 A/duck/Parung/IPB8-RS/2006 (H5N1) Parung

9 A/duck/Parung/IPB9-RS/2006 (H5N1) Parung

10 A/duck/Bojonggenteng/IPB10-RS/2006 (H5N1) Bojonggenteng

11 A/muscovy duck/Cidahu/IPB11-RS/2006 (H5N1) Cidahu

12 A/duck/Ciseeng/IPB12-RS/2006 (H5N1) Ciseeng



15 A/duck/Cileungsi/IPB15-RS/2006 (H5N1) Cileungsi

16 A/duck/Klapanunggal/IPB16-RS/2006 (H5N1) Klapanunggal



19 A/muscovy duck/Cibinong/IPB19-RS/2006 (H5N1) Cibinong

20 A/muscovy duck/Parung/IPB20-RS/2006 (H5N1) Parung

21 A/goose/Parung/IPB21-RS/2006 (H5N1) Parung

No Isolat Titik pemotongan Patotipe

1 A/muscovy duck/Klapanunggal/IPB1-RS/2006 (H5N1) QRERRRKKR/G HPAI

2 A/goose/Bojonggenteng/IPB2-RS/2006 (H5N1) QRERRRKKR/G HPAI

3 A/duck/Leuwiliang/IPB3-RS/2006 (H5N1) QRERRRKKR/G HPAI

4 A/goose/Leuwiliang/IPB4-RS/2006 (H5N1) QRERRRKKR/G HPAI

5 A/muscovy duck/Cileungsi/IPB5-RS/2006 (H5N1) QRERRRKKR/G HPAI

6 A/duck/Nagrak/IPB6-RS/2006 (H5N1) QRERRRKKR/G HPAI

7 A/goose/Klapanunggal/IPB7-RS/2006 (H5N1) QRERRRKKR/G HPAI

8 A/duck/Parung/IPB8-RS/2006 (H5N1) QRERRRKKR/G HPAI

9 A/duck/Parung/IPB9-RS/2006 (H5N1) QRERRRKKR/G HPAI

10 A/duck/Bojonggenteng/IPB10-RS/2006 (H5N1) QRESRRKKR/G HPAI

11 A/muscovy duck/Cidahu/IPB11-RS/2006 (H5N1) QRERRRKKR/G HPAI

12 A/duck/Ciseeng/IPB12-RS/2006 (H5N1) QRERRRKKR/G HPAI



15 A/duck/Cileungsi/IPB15-RS/2006 (H5N1) QRERRRKKR/G HPAI

16 A/duck/Klapanunggal/IPB16-RS/2006 (H5N1) QRERRRKKR/G HPAI



19 A/muscovy duck/Cibinong/IPB19-RS/2006 (H5N1) QRERRRKKR/G HPAI

20 A/muscovy duck/Parung/IPB20-RS/2006 (H5N1) QRERRRKKR/G HPAI

21 A/goose/Parung/IPB21-RS/2006 (H5N1) QRERRRKKR/G HPAI

Pathotype of H5N1 virus isolate from water fowl in West

Java Indonesia

The H5N1 virus could be isolated from healthy ducks

Isolate: FKH/IPB/Duck/NG26 dan

FKH/IPB/Duck/NG29, FKH/IPB/Duck/KN18 dan isolat FKH/IPB/Duck/LE16B.

The present of AI virus in the intestine of healthy duck

AMPLICON OF H5N1 AI VIRUS WITH RT-PCR

1. MARKER2. FKH/IPB/DUCK/NG293. FKH/IPB/DUCK/NG264. NOT DETECTED (-)5. FKH/IPB/CHICK/TASIK (ref)

1 2 3 4 5•PRIMER 591 bp and 237 bp were also used

1000 bp

How was the viral prevalence

in backyard chicken?

Sample Size

1

2

3

4

5

6

Kab Pandeglang

Kab Lebak

Kab Tangerang

Kab Serang

Kota Tangerang

Kota Cilegon

390

520

470

460

35

35

25

15

80

70

5

5

No. Kab/Kota Ayam Buras Itik

1.910 200Total

Seroprevalence of AI among poultry

22

17

200

5

10

15

20

25

Ayam Entog Bebek Angsa

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7°00' 7°00'

6°30' 6°30'

6°00' 6°00'

105°00'

105°00'

105°30'

105°30'

106°00'

106°00'

106°30'

106°30'

PETA TEMATIK

PREVALENSI SEROLOGIS AVIAN INFLUENZA

PROPINSI BANTENSumber Data :

Survei Seroepidemiologi Avian Influenza

Oktober – November 2006, FKH - IPB

Simbol UnggasKETERANGAN :

FAKULTAS KEDOKTERAN HEWAN

INSTITUT PERTANIAN BOGOR

33.95 %

10.71 %

0 %

0 %

KAB. LEBAK

1.47 %

0 %

0 %

0 %

KAB. PANDEGLANG

10.78 %

3.12 %

0 %

0 %

KAB. SERANG

39.33 %

30.34 %

8.33 %

0 %

KAB. TANGERANG

2.86 %

0 %

0 %

0 %

KOTA

TANGERANG

0 %

0 %

0 %

0 %

KOTA CILEGON

PROPINSI

JAWA BARAT

PROPINSI

DKI JAKARTA

PROPINSI

LAMPUNG

Ayam

Entok

Itik

Angsa

S A M U D E R A H I N D I A

LAUT JAWA

Teluk Banten

U

Lokasi contoh

Detection of Genetic Material of AI Virus

Amplicon of AI virus with RT-PCR

H5 N1

Districts Sub district

RT-PCR Results

Vaccinated Non-Vaccinated

HI+ HI- HI+ HI-

Kab. Tangerang Kronjo H5 (1)

Mauk H5 (1)

Paku Haji H5 (1) H5 (1) H5 (2)

Kresek H5 (1)H5N1 (1)

H5 (1)

Kab. Serang Carenang H5 (2) H5 (1) H5 (1)

Kragilan H5 (1)

Petir H5 (2)

Kab. Lebak Cibadak H5 (1) H5 (1) H5 (2)

G Kencana H5 (1)H5N1 (1)

H5 (1)

Muncang H5 (1)

Bayah H5 (2)

Kab. Pandeglang Cikeusik H5N1 (1)

Panimbang H5N1 (1)

Kab. Cilegon Jombang H5 (1)

Cilegon H5N1 (1)

The Year of 2009-2010

Poultry Movements

Virus affecting both humans and local poultry may not be locally produced,

but instead introduced from outside sources

New outbreak: clinical cases

in Sukabumi West Java

Desember 2009-Mei 2010

The symptoms

• Serious haemorhagic symptoms

• Sudden death, within 4-10 hours

• No more blue comb is found

• No more “Kaki kerokan” is found

Intensive haemorhagic symptom

Per-acute infection: 5-10 hours

Haemorhage of ovary

Haemorhagic symptom of adipose tissue

Haemorhagic appearance in

adipose tissue

“sudden-death”

HPAI Gross Lesions

Deteksi material genetik H5

Detection of genetic material of H5

Amplification for sequencing

0.01

Ck/West Java/TASIK2/2006

Ck/Indonesia/Siak1631-2/2006

Ck/West Java/TASIKSOL/2006

Ck/SMI-FKH IPB/2009

Ck/West Java/SMI-ENDRI1/2006

Ck/West Java/SMI-PAT/2006

Ck/West Java/SMI-CSLK-EC/2006

Ck/West Java/PWT-WIJ/2006

Ck/Purworejo/BBVW/2005

quail/Yogjakarta/UT1023/2004

Quail/Central Java/SMRG/2006

Ck/Salatiga/BBVet-I/2005

Ck/Indonesia/Wates77/2005

Ck/East Kalimantan/UT1035/2004

Ck/Purwakarta/BBVet-IV/2004

Ck/Indonesia/11/2003

Ck/Legok/2003

Ck/Indonesia/2A/2003

Ck/West Java/HAMD/2006

Ck/Bantul/BBVet-I/2005

Ck/Gunung Kidal/BBVW/2005


Ck/Magetan/BBVW/2005


Ck/East Java/UT6044/2007

Ck/Banten/UT6025/2006

Ck/Way Kanan/BBPVIII/2006

Ck/Sembawa/BPPV-III/2005

Ck/East Java/UT6021/2006

Ck/Bali/UT2091/2005

Ck/Wajo/BBVM/2005

Ck/Sulawesi Selatan/UT2093/2005

Ck/Papua/TB15/2006

Ck/Papua/TA5/2006

Source of data

• Isolates from sector 1, 2, dan 3

–Sequencing HA-1

• Deposited cDNA sequences

• Riset Unggulan Insentif

(Menristek)

WJ 6/08

WJ 7/08

WJ 5/08

West Java/PWT-WIJ/2006

EJ 1/08

EJ-2/08

WJ 4/08

Dk/Parung/IPB9-RS/06

Gs/Klapanunggal/IPB7-RS/06

Dk/Parung/IPB8-RS/06

NS-1/08

I. A virus A/Chicken/Wajo/BBVM/2005 HA

I. A virus A/duck/Parepare/BBVM/2005 HA

I. A virus (A/chicken/Gunung A/chicke...

I. A virus A/chicken/Magetan/BBVW/200...

I. A virus A/chicken/Purworejo/BBVW/2...

I. A virus A/Chicken/Yogjakarta/BBVet...

I. A virus A/quail/Yogjakarta/BBVet-I...

I. A virus A/quail/Boyolali/BPPV4/200...

WJ 3/07

I. A virus A/turkey/Kedaton/BPPV3/200...

I. A virus A/Chicken/Salatiga/BBVet-I...

I. A virus A/chicken/Pangkalpinang/BP...

I. A virus A/chicken/Dairi/BPPVI/2005 HA

I. A virus A/chicken/Simalanggang/BPP...

I. A virus A/chicken/Tarutung/BPPVI/2...

I. A virus (A/chicken/Tebing A/chicke...

I. A virus (A/chicken/Deli A/chicken/...

I. A virus A/chicken/Malang/BBVet-IV/...

I. A virus A/chicken/Purwakarta/BBVet...

Dk/Nagrak/IPB6-RS/06

Gs/Bojonggenteng/IPB2-RS/06

Md/Cileungsi/IPB5-RS/06

Md/Klapanunggal/IPB1-RS/06

Dk/Leuwiliang/IPB3-RS/06

Gs/Leuwiliang/IPB4-RS/06

I. A virus A/chicken/Pekalongan/BPPV4...

I. A virus A/Chicken/Wonosobo/BPPV4/2...

I. A virus A/quail/Tasikmalaya/BPPV4/...

I. A virus A/chicken/Ngawi/BPPV4/2004 HA

I. A virus A/Ck/Indonesia/BL/2003 HA

I. A virus A/chicken/Sragen/BPPV4/200...

I. A virus A/Ck/Indonesia/2A/2003 HA

I. A virus A/Dk/Indonesia/MS/2004 HA

I. A virus (A/chicken/Kulon A/Chicken...

I. A virus A/chicken/Kupang-1-NTT/BPP...

I. A virus A/Ck/Indonesia/5/2004 HA

I. A virus A/chicken/Jembrana/BPPV6/2...

I. A virus (A/chicken/Bangli A/chicke...


I. A virus A/chicken/Mangarai-NTT/BPP...


I. A virus A/Ck/Indonesia/4/2004 HA

VACCINE SEED/03

I. A virus A/Ck/Indonesia/PA/2003 HA

9775

97

3259

78

100

77

78

5799

90

97

96

83

93

72

79

99

100

74

4164

46

96

65

82

40

62

78

39

59

24

35

27

35

2

3

189

1

38

1931

23

4

12

44

0.005

Vaksin GMT dengan Antigen

A/Dps/04 A/Jbr/06 A/Klk/06

A/Dps/04

6.7 5.8 4.6 A/Jbr/06

6.9 8.3 4.3 A/Klk/06

5.4 5.7 6.4

Phylogenetic and

Antigenic Structure (Mahardika, 2009)

N

o

Isolat Spesies

hospes

Kelompok

(Klaster,Subklaster)

Titer

HI**

Virus Legok

2003

Ayam - 26

1 IPB1-RS Entok (2,2) 23

2 IPB2-RS Angsa (2,2) 20

3 IPB3-RS Itik (2,2) 21

4 IPB4-RS Angsa (2,2) 20

5 IPB5-RS Entok (2,2) 23

6 IPB6-RS Itik (2,1) 23

7 IPB7-RS Angsa (1) 20

8 IPB8-RS Itik (1) 20

9 IPB9-RS Itik (1) 20

No Isolat Tahun Kelompok

(Klaster,Subklaster)

Titer HI**

Virus Legok 2003 - 26

1 IPB1-RS 2006 (2,2) 23

2 IPB2-RS 2006 (2,2) 20

3 IPB3-RS 2006 (2,2) 21

4 IPB4-RS 2006 (2,2) 20

5 IPB5-RS 2006 (2,2) 23

6 IPB6-RS 2006 (2,1) 23

7 IPB7-RS 2006 (1) 20

8 IPB8-RS 2006 (1) 20

9 FKH-IPB 2009 20

** Uji HI dilakukan 2x

Cross Reaction

The use of “cock-tail” vaccine for

Avian Influenza: The vaccine consist

of 2 or 3 different AI isolates

The incidence of AI in the

commercial chicken in the

field is decrease significantly

The sporadic AI cases is still oserved

in back yard chicken

What is going to be done to

protect the “Ayam Kampung”

from AI out-break?

Conclussion

The form of “SUBCLINICAL INFECTION” might play a significant and important role in the spreading of the disease and might act as source of infection to other species

The new outbreak of AI in the last of 2009 and the beginning of 2010 is caused by new “strain” of AI virus which has no cross reaction with specific antibody to strain of 2003 (Legok)

FACULTY OF VETERINARY MEDICINE BOGOR …zoonosis.ipb.ac.id/pdf_file/zoonosis centre 19 April 2012-wayan-1.pdf · FACULTY OF VETERINARY MEDICINE BOGOR AGRICULTURAL UNIVERSITY, BOGOR.

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