1. ANTIGENIC AND EVOLUTIONARY ANALYSIS OF FMD VIRUSES FROM THE 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. O Ind-2001d in Lybia from 2013 Introduction
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

3. Objective
Get insight virus evolution and epidemiological relationship between FMD viruses from 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. Materials and methods
Miseq Istrument (Illumina) SeqMan Ngen 12.0 (DNASTAR)
Full genome sequence of 8076 nucleotides
PHYLOGENETIC ANALYSIS
7,5-8,5 Kb
ORF/Polyprotein
55 MAbs
22 neutralizing and 23 non neutralizing
ANTIGENIC PROFILE

5. Phylogenetic analysis (VP1)
Bayesian evolutionary analysis (BEAST)
LIBYA
TUNISIA
ALGERIA
MOROCCO
2013
2014
2015
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

6. 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 in Algeria
Tunisian field viruses clustered in three main branches
TUNISIA
ALGERIA
2014
2015

7. Phylogenetic analysis (ORF)
Statistical parsimony analysis as implemented by TCS
Neutralizing
Site 1
Neutralizing
Site 2
Neutralizing
Site 1
Neutralizing
Site 4
07/05/14
09/06/14
26/04/14
15/09/14
01/06/14
14/05/14
05/09/14
14/07/14
28/05/14
29/07/14
07/07/14
29/07/14
20/09/14
Nucleotide substitutions
Tunisia isolates 2014
Algerian isolates 2014
Nonsynounymous substitutions on SP
Nonsynounymous substitutions on NSP
a total of 48
in twelve isolates
High number of nucleotide substitutions between and within clusters

8. Antigenic profile by monoclonal antibodies
0-14
15-39
40-69
70-100
Monoclonal
Antibodies
O LIB B5 1/8/13
O LIB E1 8/9/13
O LIB Y3 22/10/13
O TUN /4/14
O TUN /5/14
O TUN /5/14
O TUN /7/14
O TUN /5/14
O TUN /5/14
O TUN /6/14
O TUN /6/14
O TUN /7/14
O TUN 2152 B 5/9/14
O TUN /9/14
O ALG 1628 Jul 14
O ALG 1631 Jul 14
O Manisa
O BFS
O UK2001 (PanAsia)
NEUTRALIZING MAbs
7E1
2C10
2F10
1E9
4C9
4C6
4E3 B2
2A10
1D7
4E1
3B12
1C12
4B7
1C6
3H4
1F3
2E9
2G6
1A7
3D8
6C3
Monoclonal Antibodies
O LIB B5 1/8/13
O LIB E1 8/9/13
O LIB Y3 22/10/13
O TUN /4/14
O TUN /5/14
O TUN /5/14
O TUN /7/14
O TUN /5/14
O TUN /5/14
O TUN /6/14
O TUN /6/14
O TUN /7/14
O TUN 2152 B 5/9/14
O TUN /9/14
O ALG 1628 Jul 14
O ALG 1631 Jul 14
O Manisa
O BFS
O UK2001 (PanAsia)
NON NEUTRALIZING MAbs
2D2
1F7
1H3 A8
2B8
2B1
1F12
3F5
1B1
3C9
1H10
2E3
3D7
4H12
3C8
1D5
1B3
1G5
1G3
3B11
1E10
4D11
Target site 1
Target site 2
Target site 3
Mixed Target sites

9. Two different FMDV introductions in Algeria
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. THANK YOU FOR YOUR ATTENTION