“control of exotic and emerging animal diseases”

1. Research on heartwater and TBTCIRAD Guadeloupe Development of integrated control against the tick-borne diseases • OIE Reference laboratory : Heartwater • Coordination of global network of research • PCRD5 and 6 and 7? • 5 European countries, 7 African and 2 American • Biology, genomics, diagnostic, immunology, control of ER and TBT Generation of diversity and evolution of microbial genomes as factors for emerging, spreading and virulence determination Mechanisms for host response to pathogens and vectors, development of control methods Animal health network Population dynamic & genetic of vectors, disease transmission dynamics. Modeling for risk mapping and control strategies “control of exotic and emerging animal diseases”

2. Ehrlichia ruminantium functional genomic SCOTS 0x SCOTS 3x gDNA RM 96Hpi significativity variance • Genomic • Genome publications & patents: 2 strains of Ehrlichia ruminantium Sequencing & annotation: 3 additional strains virulent/attenuate • development of tools for diagnostic and characterisation • comparative genomic and functional genomic • Transcriptomic • Comparison between virulent and attenuate strains • Comparison between stages of development • ER = Obligate intracellular pathogen  Adaptation of SCOTS method to capture bacterial transcripts •  Microarrays and qRT-PCR validation: • ~30 genes discriminate virulent/attenuate strains • genes differentially expressed or presence/absence  Identification of pathogenesis mechanisms • virulence & adaptation to host cells, attenuation. •  Development of efficient recombinant vaccines Emboulé et al (Submited) BMC Molecular biology Frutos et al, Trends Parasitol, 2007 Frutos R, et al, J Bacteriol, 2006

3. Diversity of Ehrlichia ruminantium,new diagnostic tools and prevalence RU-3 RU-7 RU-2 • Genetic characterization of diversity: MAP Multi genic family • MAP1 good tool for typing: 80 strains Afrique/O.Indien/Caraïbes • High diversity in restricted areas • Identification of genes correlated with protection • Multi Locus Sequence Analysis (MLSA & MLST) & • Variable Number of Tandem Repeats (VNTR) • New molecular tools for large epidemiological studies: 3 areas • Optimization of tools • MLST: on 7 housekeeping genes • MLSA: on 10 polymorphic genes • VNTR: 17 loci identified • = Phylogeny/ Phylogeography at high resolution/ Structural study of populations • Ehrlichia ruminantium infection level in TBTs • Guadeloupe 16% Marie Galante 19% Antigua 6% • Burkina Faso (Banan) 10% • Madagascar low prevalence / Comores: 20-30% Adakal et al Infect Genet Evol, 2009 Raliniaina et al, Veterinary Parasitology, 2009 Vachiéry et al, Infect Genet Evol, 2008

4. Recombinant vaccine against Ehrlichia ruminantium mapX7 mapX8 mapX12 map1-2 map1 mapX11 • Identification of candidate genes: sequencing, annotation • 3D MAP proteins structure determination (RMN): • Conformational studies • Identification of epitopes of interest • Vaccine assays : • 50% of protection with a mix of 4 MAP proteins • improvement of protein expression (purity, quantity, solubility) • vaccine assays with individual recombinant proteins or specific epitopes 3D predictive model of MapX12 Erga vs. Erwe

5. Inactivated vaccine against Ehrlichia ruminantium (IBET+CIRAD) • Determination of efficient vaccine dose • In vivo assay on goat: ↓vaccine dose from 1mg to 35µgr • Scaling up from static conditions to stirred tanks: Stirring / Medium • Cost decrease : 3600 doses for 2 liters 0.11 €/dose • Optimization of antigen preparation • Adaptation to industrial process: filtration • Gain Time/ ER Yield: 45 min and 8 fold ER recovery yield • Test of buffers and storage conditions: NaCl/PBS -20°C/4°C • In vivo assay : Validation of conditions: NaCl/-20°C • Formulation of the vaccine: Vaccine ready-to-use • Preparation Adjuvant + Antigene emulsified • Length of storage • In vivo Assay on goat* Efficient vaccine with cost effective process: Adaptation to any strain Possible application to similar biological systems: Chlamydiae, Rickettsie Marcelino et al, Vaccine, 2007 Vachiéry et al, Vaccine, 2006

6. TBTs infestation level in the Caribbean Nevis • French territories • Guadeloupe (farms 36%), Marie Galante (74%), Martinique (4%) • CAP countries, TickINFO • Drop of prevalence induced by treatment • Risk factors: 3rd quarter and cattle • Resurgence • St Kitts: re-infestation • Barbados/Nevis: few hotspots • St Lucia: outbreaks • Need for random continues surveillance data? • Importance of farm factors: lack of information about herd management, treatment compliance, area … • TBT genetics studies to explain variable infestation levels • Barbados • St Kitts St Lucia Nevis. Ahoussou et al, Veterinary Parasitology, 2009 Molia et al, Vet Parasitol, 2008 Vachiéry et al, Ann N Y Acad Sci. 2008

7. Modelling TBT population dynamics • Mathematical model • TBT Biology and ecology (development, mortality, host finding, laying) • Biotic et abiotic factors • Comparison to known data • Risk based surveillance • Test control efficiency • Spreading potential • Habitat suitability model • Humidity, temperature, photoperiod

8. Research on tick & tick borne disease:Amblyomma variegatum (TBT) & Ehrlichia ruminantium (ER) ER biology genetics, genomics Vector competence & vector capacity TBT biology and ecology TBT population genetics ER Diversity studies Disease modeling Emerging factors Spreading risk Control strategies ER population dynamics TBT population dynamics ER genetic resistance? ER vaccine development Host response to TBT