ANTIGENIC AND EVOLUTIONARY ANALYSIS OF FMD VIRUSES FROM THE 2014-2015 OUTBREAKS IN THE MAGHREB REGION G. Pezzoni1, M. Calzolari1, S. Sghaier2, H. Madani3,

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Presentation transcript:

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)

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

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

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

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

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 TUNISIA ALGERIA 2014 2015

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

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 1030 26/4/14 O TUN 1101 5/5/14 O TUN 1114 7/5/14 O TUN 1846 15/7/14 O TUN 1223 14/5/14 O TUN 1412 28/5/14 O TUN 1771 1/6/14 O TUN 1566 9/6/14 O TUN 1797 7/7/14 O TUN 2152 B 5/9/14 O TUN 2157 15/9/14 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 1030 26/4/14 O TUN 1101 5/5/14 O TUN 1114 7/5/14 O TUN 1846 15/7/14 O TUN 1223 14/5/14 O TUN 1412 28/5/14 O TUN 1771 1/6/14 O TUN 1566 9/6/14 O TUN 1797 7/7/14 O TUN 2152 B 5/9/14 O TUN 2157 15/9/14 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

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

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