This study analyzed 55 foot-and-mouth disease virus (FMDV) isolates from Turkey between 1999-2020 to monitor the evolution of serotype O. Two genotypes were identified: O/ME-SA/Panasia-1 circulated 1999-2006, and O/ME-SA/Panasia-2 has circulated since 2006 with six subtypes. Comparison of amino acid sequences found changes between genotypes and subtypes, including some in the antigenic G-H loop region. Endemic countries need to share timely data to understand outbreak dynamics and reduce future risks. Vaccine strains were updated to match circulating strains.

1. MONITORING TO COMBAT THE FOOT AND MOUTH DISEASE VIRUS
SEROTYPE O FROM 1999 TO THE PRESENT IN TURKEY
Sena İNEL TURGUT *1, Pelin TUNCER-GÖKTUNA2, Ünal PARLAK1,
Fuat ÖZYÖRÜK3, Gül Nazik BALCI1, Abdurrahman KURT1, Sibel YAVRU4
1Foot-and-Mouth Disease (SAP) Institute, Ankara, Turkey
2İstanbul Pendik Veterinary Control Institute, Istanbul, Turkey
3Harran University, Faculty of Veterinary Medicine, Department of Virology, Sanlıurfa, Turkey
4Selcuk University, Faculty of Veterinary Medicine, Department of Virology, Konya, Turkey
INTRODUCTION
Foot-and-mouth disease virus (FMDV) causes one of the most devastating disease
in the cloven-hoofed animals. It is belongs to the Apthovirus genus in the
Picornaviridae family. It has got seven serotypes (A, O, C, SAT1, SAT2, SAT3, and
Asia-1) and the serotypes are grouped in seven pools according to the geographic
distributions (1). Turkey is in pool three (A, O, Asia-1) and serotype O is accepted
as local strain while other two are accepted as exotic strains (2). Like the other RNA
viruses, FMDV mutating with a very high rate, there is no cross-protection between
serotypes. Sometimes protection may not be provided even among different
subtypes of the same serotypes (3). Therefore, the evolution of serotypes must be
closely monitored to combat with the FMDV.
MATERIALS AND METHODS
Fifty-five isolates from serotype O and the vaccine strains were selected and
studied by using sequenced viral protein 1 (VP1) by sanger sequencing.
Phylogenetic relationships constructed with minimum-evolution method by using
MEGA 6.06 software. Kimura 2-parameter nucleotide substitution model
implemented and bootstrap analysis was undertaken using 2500 replicates of the
dataset.
RESULTS
As seen in a phylogenetic tree (Figure 1), two different Panasia and one non-
panasia(CII) genotypes have been identified. First one is O/ME-SA/Panasia-1
genotype. It was circulated with two subtypes between the years 1999-2006 (α, β)
(Table 1). The second one is, O/ME-SA/Panasia-2 genotype. It has been detected
since 2006 (Table 1-2) in the field with its six subtypes; I(USK-06), II, TER-08, FAR-
09, ANT-10, QOM-15, respectively. Besides, O/ME-SA/Iran-2001 genotype
circulated with O/ME-SA/Panasia-1 genotype between the years 2000-2004
(except 2003) (Table 1).
According to an analysis of the VP1 regions of two panasia genotypes, pairwise
distance obtained as %9. There were seven different amino acids detected
between the first Panasia-2 isolates and the last Panasia-1 isolates. (Table 2).
Three of them (137,140,141) were in the G-H loop of VP1 [RGD (Arg-Gly-Asp)]
which is critical in the antigenicity of FMDV (4).
REFERENCES
1. OIE. 2012. Chapter 2.1.5. Foot and mouth disease. In Manual of standards for diagnostic tests and vaccines for
terrestrial animals. OIE, Paris, France.
2. Parlak Ü, Özyörük F, Knowles NJ, Armstrong RM, Aktas S, Alkan F, Cokcaliskan C, Christensen LS. 2007.
Characterisation of foot-and-mouth disease virus strains circulating in Turkey during 1996-2004. Archieves of
Virology, 152: 1175-1185.
3. Parida S. 2009. Vaccination against foot-and-mouth disease virus: strategies and effectiveness. Expert Rev
Vaccie, 8: 347-365.
4. Grubman MJ, Baxt B. 2004. Foot-and-mouth disease. Clinical Microbiology Reviews, 17: 465-493.
5. Sap Instıtute, Diagnosis Dept,Typing and Moleculer Epidemiology Lab archive.
6. Knowles NJ, Samuel AR, Davies PR, Midgley RJ, Valarcher JF. Pandemic strain of foot-and-mouth disease virüs
serotype O. EmergInfectDis. 2005;11(12):1887-1893. doi:10.3201/eid1112.050908
7. OIE-FAO FMD Ref Lab Report Oct-Dec 2006.
8. OIE-FAO FMD Ref Lab Report Jan-Mar 2007.
Table 1. The lineages circulating during 1999-2020 in Turkey(5)
Years of detection
Lineage
Sub-
Lineage
1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017 2018 2019 2020
non-PA
(CII) Iran2001
O/ME-
SA/Pa-1 α
β
O/ME-
SA/Pa-2
I(USK-06)-II
TER-08
ANT-10
FAR-09
QOM-15
Figure 1. Phylogenetic analysis of VP1-coding sequences of serotype O
Bootstrap values >%80 are indicated.
*Since the chromatograms of most of the samples before 1999 (Iran 2001) were not suitable, samples were selected
for analysis from previous years.
O/Erzurum/287/2008.808
O/Rize/290/2008.846
O/Aksaray/178/2009.596
O/Sanliurfa/264/2009.904
O/Erzincan/257/2008.615
O/Nevsehir/40/2009.192
O/Erzincan/258/2009.808
TER-08
O/Sirnak/285/2007.135
O/Sivas/8/2008.038
O/Konya/120/2007.077
O/Usak/1423/2006.788
O/Kirklareli/357/2007.173
I-USK-06
O/Ardahan/57/2015.231
O/Agri/63/2013.115
O/Nigde/49/2014.077
O/Samsun/129/2010.212
O/Agri/71/2010.096
O/Eskisehir/205/2012.192
O/Bayburt/1251/2012.962
FAR-09
O/Gaziantep/153/2010.212
O/Ankara/86/2011.019
O/Isparta/54/2011.000
O/Corum/1428/2011.596
O/Tokat/178/2020.750
O/Van/156/2020.635
O/Izmir/427/2017.942
O/Antalya/428/2017.942
ANT-10
O/Sakarya/437/2017.962
O/Bingol/370/2017.769
O/Ankara/504/2018
O/Van/445/2018.558
O/Kars/96/2016.038
O/Tokat/289/2017.519
O/Zonguldak/131/2019.942
O/Izmir/188/2020.769
O/Ardahan/5/2020.038
O/Elazig/136/2020.385
O/Balikesir/7/2019.096
O/Yozgat/189/2020.769
O/Artvin/165/2020.712
O/Konya/137/2020.404
O/Malatya/147/2019.1000
O/Samsun/55/2020.115
QOM-15
Panasia 2
O1/Manisa/TUR/1969.250*
O/TUR/3/1987.404*
O/Mersin/35/2004.058
O/TUR/2/2000
Iran-2001
O/Ankara/TUR/643/1999.962
O/Erzincan/TUR/351/2002.788
O/Konya/TUR/512/10/1999.769
ß
O/Ankara/TUR/250/2001.231
O/Denizli/TUR/87/2003.173
O/KMaras/1127/2006.500
O/Samsun/336/2004.999
O/Erzurum/423/2005.558
α
Panasia 1
100
100
100
82
56
99
99
52
55
50
94
93
69
50
41
52
21
73
65
69
100
79
72
91
74
54
99
67
70
64
96
68
82
99
86
41
70
86
66
36
45
93
80
77
61
97
60
38
56
32
99
28
0,01
Position Panasia-1 Panasia-2 Position O1 Manisa O/TUR/07 Position USK-06 QOM-15
43 T S 4 A T 45 K R
102 A T 96 A T 96 T M
137 A V 138 D E 133 N D
140 Q H 139 G S 140 H R
141 A V 140 T H 141 T A
195 S V 141 V T 195 H Q
195 P N 142 A T 197 D S
158 A T 209 V G
198 Q E
Table 2. Amino acid differences observed at VP1
CONCLUSION
O/ME-SA/Panasia-1 genotype, which began in 1998 in South Asia and led to
the pandemic, was identified in 1999 in Turkey (6). It has been noted that the
O/ME-SA/Panasia-2 outbreaks that began in Iran and Turkey were an
extension of Jordan (7). The spread may be due to the introduction of new live
animals, illegal movement of animals, and contact with infected animals (8).
Endemic countries located in the same pool need to exchange data on
time and act simultaneously. It will affect the dynamics of the outbreak
and reduce the risks associated with the future of the outbreaks.
II
During the O/ME-SA/Panasia-1 genotype circulation, O1 Manisa vaccine strain was
included in vaccines. After incursion of the O/ME-SA/Panasia-2 genotype with high
mortality, O/TUR/07 vaccine strain was produced in 2007. There are nine different
amino acids identified between the vaccine strains and five of them
(138,139,140,141,142) are close to G-H loop of VP1 [RGD (Arg-Gly-Asp)] (Table 2).
Also, some changes have occurred in eight amino acids between the USK-06, and
the QOM-15 subtypes (the first and the last isolates of O/ME-SA/Panasia 2). Three
of them (133,140,141) are close to the same antigenic site; the G-H loop (Table 2).
Nevertheless, O/TUR/07 still works according to r1-tests and in the field. It is still
used as one of the main vaccine strains in Turkey.