1. Department of Biotechnology University of Malakand, Pakistan
Genetic Characterization of foot-and-mouth disease viruses in Balochistan, Pakistan
Syed M. Jamal
Department of Biotechnology University of Malakand, Pakistan

2. 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. Balochistan Province of Pakistan
Southwestern part of the country
The largest province shares borders with Afghanistan (1100 km) and Iran (909 km)
The geographical location of this Province
Animal Trade among Pakistan, Iran and Afghanistan
spread of transboundary animal diseases including FMD
Afghanistan
Thus geographical location of this Province has importance in both formal and informal trade of animals among Pakistan, Iran and Afghanistan and spread of transboundary animal diseases including FMD.
Balochistan shares Border with Afghanistan (~1100 km) total =2430 km)
Balocihstan shares border with Iran 909 km
India
Iran
Arabian sea

4. Methodology 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

5. Results Assay Original samples Tissue culture Total Positive Ag-ELISA
A/As = 1
Assay
Original samples
Tissue culture
Total
Positive
Ag-ELISA 35 8
Pan-FMDV rRT-PCR 31
Serotype-specific rRT-PCR 29
A = 16
As = 8
A/As= 5
No sample positive for serotype O

6. ML tree generated using VP1 coding sequences of serotype A FMDV
Pakistan & Afghanistan 2009
FAR-11
ML tree generated using VP1 coding sequences of serotype A FMDV
A-Iran05
BAL-11
New sublineage
FAR-11 show close identity with viruses circulating in Pakistan and Afghanistan in 2009
HER-10 show close identity with viruses circulating in Kirgizstan in 2011 and Iran and Kazakhstan in 2012
HER-10
Afghanistan 2010
Kirgizstan 2011
Iran 2012
Kazakhstan 2012

7. ML tree generated using VP1 coding sequences of serotype Asia-1 FMDV
First detected in 2008 in samples originated in southern par t of the country

8. Conclusions
Different sublineages within A-Iran05 are in circulation and new sublineages arise with time.
Detection of new strain of serotype A suggests that continuous surveillance of the disease is necessary
Oral swab samples can be used for virus isolation and subsequent characterization
Collection of oral swab samples is non invasive compared to the epithelial samples and therefore acceptable to the animal owners
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. Acknowledgements Asad Ulllah, CASVAB, University of Balochistan
Stephan Zientara and Labib Kassimi and their team
EuFMD team
Higher Education Commission, Islamabad, Pakistan
GFRA