1.
Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
Syed M. Jamal
Department of Biotechnology
University of Malakand, Pakistan
Genetic Characterization of foot-and-mouth
disease viruses in Balochistan, Pakistan

2.
OS16
FMD in Pakistan
Endemic country
Three serotypes (O, A, Asia-1)
Multiple subtypes
Studies on genetic diversity of FMDVs in Pakistan (Jamal et al 2011a,b,c; Waheed et
al., 2012; Jamal et al., 2012; Brito et al., 2013)
However, similar information on FMDV in Balochistan province of Pakistan is not
available.
Objectives of the study was to characterize FMDVs in Balochistan

3.
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o Southwestern part of the country
o The largest province shares borders
with Afghanistan (1100 km) and Iran
(909 km)
o The geographical location of this
Province
o Animal Trade among Pakistan,
Iran and Afghanistan
o spread of transboundary animal
diseases including FMD
Afghanistan
Iran
Arabian sea
India
Balochistan Province of Pakistan

4.
OS16
Number of samples collected = 35
Epithelial tissues = 2
Oral swabs = 33
Number of locations = 8
Transported to ANSES, Laboratory for Animal Health, France
Analysis of samples
 Ag-ELISA
 Cell culture (foetal goat tongue cell line, ZZ-R 127) & Ag-ELISA
 Pan-FMDV 3D assay
 Serotype-specific qRT-PCR assays
 Sequencing
Methodology

5.
Results
Assay Original samples Tissue culture
Total Positive Total Positive
Ag-ELISA 35 0 8 8
Pan-FMDV rRT-
PCR
35 31 8 8
Serotype-specific
rRT-PCR
35 29 8 8
A = 5
As = 2
A/As = 1
No sample positive for serotype O

6.
OS16
FAR-11
BAL-11
HER-10
A-Iran05
HIR/AFG/L1575/2009 (HQ439291)
PUN/PAK/L2808/2009 (HQ439257)
TAK/AFG/L1420/2009 (HQ439264)
PUN/PAK/L1354/2009 (HQ439251)
PAN/AFG/L3641/2009 (HQ439309)
HIR/AFG/L1571/2009 (HQ439289)
A/BAL/PAK/ISO2/2011 (JX435106)*
JAW/AFG/L1439/2009 (HQ439278)
GU384686 (A/PAK/76/2009)
SIN/PAK/L574/2009 (HQ439244)
SIN/PAK/L578/2009 (HQ439245)
HIR/AFG/L1486/2009 (HQ439283)
SAR/AFG/L1435/2009 (HQ439274)
FJ755009 (AFG/44/2007)
FJ755007 (AFG/6/2007)
AFG-07
SAR/AFG/L1434/2009 (HQ439273)
KAB/AFG/L1163/2008 (HQ439262)
SIN/PAK/L695/2009 (HQ439248)
SIN/PAK/L693/2009 (HQ439246)
FJ755010 (BAR/6/2008)
FJ755011 (BAR/7/2008)
A/BAL/PAK/23/2011 (KT832834)
A/BAL/PAK/ISO3/2011 (JX435107)
A/BAL/PAK/12/2011 (KT832827)
A/BAL/PAK/ISO4/2011 (KT832825)
A/BAL/PAK/30/2011 (KT832837)
A/BAL/PAK/34a/2011 (KT832840)*
A/BAL/PAK/19/2011 (KT832829)
A/BAL/PAK/05/2011 (KT832824)
A/BAL/PAK/21/2011 (KT832830)*
A/BAL/PAK/ISO7/2011 (KT832835)
A/BAL/PAK/ISO8/2011 (KT832836)
A/BAL/PAK/ISO5/2011 (JX435108)
A/Mainkum/02/2012 (JQ765588)
A/Kirgizia/11/2011 (JQ765587)
A/IRN/12/2012 (KT223558)
A/BAL/PAK/ISO1/2011 (JX462599)
A/BAL/PAK/22/2011 (KT832832)*
A/BAL/PAK/30a/2011 (KT832838)
A/BAL/PAK/34/2011 (KT832839)*
BAD/AFG/L3025/2009 (HQ439306)
BAD/AFG/L2815/2009 (HQ439297)
PUN/PAK/L2809/2009 (HQ439258)
PUN/PAK/L1364/2009 (HQ439253)
EU414535 (Pak/9/2003)
EU414534 (PAK/28/2002)
FJ798194 (A/PAK/73/2007)
AY593763 (A22/IRQ/64)
99
99
99
99
98
60
59
99
98
89
95
90
99
99
78
91
99
5394
88
99
81
82
99
62
65
0.02
ML tree generated using VP1 coding
sequences of serotype A FMDV
Pakistan & Afghanistan 2009
Afghanistan 2010
Kirgizstan 2011
Iran 2012
Kazakhstan 2012
New sublineage

7.
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As/BAM/AFG/L2824/2009 (HQ439204)
As/BAM/AFG/L2823/2009 (HQ439203)
As/BAM/AFG/L3/2009 (HQ439196)
As/BAM/AFG/L1/2009 (HQ439194)
As/BAM/AFG/L639/2009 (HQ439198)
As/AFG/44/2003 (EF457992)
As/KRG/1/2004 (FJ785248)
As/HKN/1/2005 (DQ121114)
As/TAJ/6/2004 (FJ785276)
As/UZB/2003 (FJ785277)
As/AFG/33/2003 (EF457990)
As/AFG/26/2003 (EF457989)
As/PAK/1/2004 (DQ121128)
As/PAK/1/2004 (HQ268513)
PAK/069/2003 (HQ268514)
As/PAK/69/2003 (DQ121127)
As/PAK/31/2002 (DQ121125)
As/PAK/030/2002 (JF749849)
As/PAK/30/2002 (DQ121124)
Group-II
As/PAK/20/2003 (DQ121126)
As/PAK/3/98 (FJ785261)
Group-VI
As1/Shamir/ISR/89
As/IRN/25/2001 (FJ785244)
As/AFG/4/2001 (DQ121110)
As/AFG/1/2001 (DQ121109)
Group-I
As/SIN/PAK/L2810/2009 (HQ439189)
As/SIN/PAK/L2812/2009 (JN006721)
As/SIN/PAK/L2954/2009 (HQ439193)
As/SIN/PAK/L5/2008 (HQ439187)
As/Slemani/2/IRQ/2014 (KM047049)
As/Slemani/IRQ/2012 (JX455113)
As/TUR/13/2013 (KM268898)
As/BAL/PAK/ISO2/2011 (JX435109)*
As/BAL/PAK/08/2011 (KT832826)
As/BAL/PAK/ISO6/2011 (KT832828)
As/BAL/PAK/21/2011 (KT832831)*
As/BAL/PAK/22/2011 (KT832833)*
As/BAL/PAK/34/2011 (KT832841)*
As/BAL/PAK/ISO1/2011 (JX435110)
Group-VII (Sindh-08)
98
99
98
81
94
97
100
88
73
98
99
86
100
99
97
89
78
85
94
90
ML tree generated using VP1 coding
sequences of serotype Asia-1 FMDV

8.
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Conclusions
o Different sublineages within A-Iran05 are in circulation and new sublineages arise
with time.
o Detection of new strain of serotype A suggests that continuous surveillance of the
disease is necessary
o Oral swab samples can be used for virus isolation and subsequent characterization
o Collection of oral swab samples is non invasive compared to the epithelial samples
and therefore acceptable to the animal owners
o Detection of two different serotypes or/and two different strains of the same
serotype in one animal/sample shows complexity of the disease in the region.

9.
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Acknowledgements
Asad Ulllah, CASVAB, University of Balochistan
Stephan Zientara and Labib Kassimi and their team
EuFMD team
Higher Education Commission, Islamabad, Pakistan
GFRA