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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
13 A/duck/Ciseeng/IPB13-RS/2006 (H5N1) Ciseeng
14 A/duck/Ciseeng/IPB14-RS/2006 (H5N1) Ciseeng
15 A/duck/Cileungsi/IPB15-RS/2006 (H5N1) Cileungsi
16 A/duck/Klapanunggal/IPB16-RS/2006 (H5N1) Klapanunggal
17 A/duck/Leuwiliang/IPB17-RS/2006 (H5N1) Leuwiliang
18 A/duck/Leuwiliang/IPB18-RS/2006 (H5N1) Leuwiliang
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
13 A/duck/Ciseeng/IPB13-RS/2006 (H5N1) QRERRRKKR/G HPAI
14 A/duck/Ciseeng/IPB14-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
17 A/duck/Leuwiliang/IPB17-RS/2006 (H5N1) QRERRRKKR/G HPAI
18 A/duck/Leuwiliang/IPB18-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/Indonesia/Wates130/2005
Ck/Magetan/BBVW/2005
Ck/Indonesia/Wates1/2005
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/Kupang-3-NTT/BPP...
I. A virus A/chicken/Mangarai-NTT/BPP...
I. A virus A/chicken/Kupang-2-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)
