1.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
ANTIGENIC AND EVOLUTIONARY ANALYSIS OF
FMD VIRUSES FROM THE 2014-2015 OUTBREAKS
IN THE MAGHREB REGION
G. Pezzoni1, M. Calzolari1, S. Sghaier2, H. Madani3, C. Chiapponi1, S. Grazioli1, A. Relmy4, L.
Bakkali4, E. Foglia1, E. Brocchi1
1. Istituto Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna (IZSLER), 2. Institut de la
Recherche Vétérinaire de Tunisie, 3. Institut National de la Médecine Vétérinaire (Algeria), 4. Agence
Nationale de Sécurité Sanitaire (ANSES)

2.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
 April 2014 to November 2015
• FMDV outbreaks in Maghreb region
• First FMDV incursion after 15 years of
absence
• FMDV lineage type O Ind-2001d based on
phylogenetic analysis of VP1 region
 Lineage type O Ind-2001d
• Belong to Middle East-South Asia topotype (ME-SA)
• First identified in Indian sub-continent in 2001
• Sporadic outbreaks in Arabian peninsula
O Ind-2001d in Lybia from 2013
Introduction

3.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
Objective
Get insight virus evolution and epidemiological relationship between
FMD viruses from 2014-2015 epidemic in Maghreb region
• 12 isolates from Tunisia from April to September 2014 epidemic
• 5 isolates from Algeria from July 2014 to February 2015 epidemic
• Phylogenetic analysis
• Antigenic profile

4.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
7,5-8,5 Kb
ORF/Polyprotein
Full genome sequence
of 8076 nucleotides
55 MAbs
22 neutralizing and
23 non neutralizing
ANTIGENIC PROFILE
Materials and methods
Miseq Istrument (Illumina)
SeqMan Ngen 12.0
(DNASTAR)
PHYLOGENETIC ANALYSIS

5.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
 Unique introduction in North Africa (all Libyan isolates share a common ancestor ͌ Jul 2013)
 From Libya, unique introduction into Maghreb region (common ancestor beginning 2014)
 Tunisian field viruses evolved on different branches
 Two different introductions into Algeria
Phylogenetic analysis (VP1) Bayesian evolutionary analysis (BEAST)
LIBYA
TUNISIA
ALGERIA
MOROCCO
20152013 2014

6.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
Phylogenetic analysis (ORF) (BEAST)
 The full genome analysis confirmed a
common ancestor between Tunisian and
Algeria 2014 isolates
 The Morocco isolate was confirmed to be
closely related to the second incursion in 2015
in Algeria
 Tunisian field viruses clustered in three main
branches
2015
ALGERIA
TUNISIA
2014

7.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
Phylogenetic analysis (ORF)
Statistical parsimony analysis as implemented by TCS
07/05/14
09/06/14
26/04/14
15/09/1401/06/14
14/05/14
05/09/14
14/07/14
28/05/14
29/07/14
07/07/14
29/07/14
20/09/14
Neutralizing
Site 4
Neutralizing
Site 1
Neutralizing
Site 1
Tunisia isolates 2014
Algerian isolates 2014
High number of nucleotide substitutions between and within clusters
Nonsynounymous substitutions on SP
Nonsynounymous substitutions on NSP a total of 48
in twelve isolates
Neutralizing
Site 2
Nucleotide substitutions

8.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
Antigenic profile by monoclonal antibodies
Monoclonal
Antibodies
OLIBB51/8/13
OLIBE18/9/13
OLIBY322/10/13
OTUN103026/4/14
OTUN11015/5/14
OTUN11147/5/14
OTUN184615/7/14
OTUN122314/5/14
OTUN141228/5/14
OTUN17711/6/14
OTUN15669/6/14
OTUN17977/7/14
OTUN2152B5/9/14
OTUN215715/9/14
OALG1628Jul14
OALG1631Jul14
OManisa
OBFS
OUK2001(PanAsia)
NEUTRALIZINGMAbs
7E1
2C10
2F10
1E9
4C9
4C6
4E3
B2
2A10
1D7
4E1
3B12
1C12
4B7
1C6
3H4
1F3
2E9
2G6
1A7
3D8
6C3
Monoclonal
Antibodies
OLIBB51/8/13
OLIBE18/9/13
OLIBY322/10/13
OTUN103026/4/14
OTUN11015/5/14
OTUN11147/5/14
OTUN184615/7/14
OTUN122314/5/14
OTUN141228/5/14
OTUN17711/6/14
OTUN15669/6/14
OTUN17977/7/14
OTUN2152B5/9/14
OTUN215715/9/14
OALG1628Jul14
OALG1631Jul14
OManisa
OBFS
OUK2001(PanAsia)
NONNEUTRALIZINGMAbs
2D2
1F7
1H3
A8
2B8
2B1
1F12
3F5
1B1
3C9
2D2
1H10
2E3
3D7
4H12
3C8
1D5
1B3
1G5
1G3
3B11
1E10
4D11
0-14
15-39
40-69
70-100
Target site 3
Target site 1
Target site 2
Mixed Target sites

9.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
CONCLUSIONS
 All Tunisian isolates shared a common putative ancestor
and evolved in different branches, probably in relation
with animal movements
 Two different FMDV introductions in Algeria
 Many nucleotide substitutions between and within
isolates: probable undetected FMDV infections
 Limited antigenic variation among isolates, restricted to
neutralizing MAbs

10.
Pezzoni Giulia - Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016