Bacillus anthracis, a spore-forming pathogenic bacterium found throughout the world, is the causative agent of anthrax. In France, a few sporadic cases are recorded each year in areas where outbreaks had been reported previously, but large outbreaks could occur. Bacillus anthracis is one of the most feared microorganisms due to its high rate of infectivity and lethality, to its global dispersal pattern and to the extreme stability of its spores. It is considered as the main threat in what concerns bioterrorism. Different strategies exist to counter this threat. This project aims at improving forensics and bio-traceability of this potential biowarfare agent, thus discouraging its intentional use. B. anthracis is considered as an evolutionarily young species, recently emerged from the B. cereus group. It presents very low molecular diversity among strains. Due to this genetic homogeneity, only a combination of genotyping methods with high discrimination power (multiple-locus variable-number tandem repeat analysis (MLVA), single nucleotide repeat (SNR), genome-wide single nucleotide polymorphism (SNP) analyses or even whole-genome sequencing) has proved to be successful in differentiating strains. The recent and dramatic improvement of whole genome sequencing technology should allow its more frequent use for systematic characterization of B. anthracis. Our project aims at improving knowledge of the existing diversity in B. anthracis isolates in France in order to develop a highly precise and robust subtyping system and know-how for the primary investigation of strain traceability. High-resolution whole-genome sequencing and comparative genomics will be applied to 25 to 50 French strains from different geographic areas to identify their key genetic features. The clonal nature of B. anthracis makes SNPs, which are scattered throughout the whole genome, ideal signatures for subtyping this pathogen. Only rare diagnostic markers can provide the level of discrimination required for microbial forensics and for precise strain identification. As a consequence of this project, France will possess the required know-how to react rapidly when a strain identity needs to be established. High-Resolution Melting (HRM) will be used to validate these potentially diagnostic SNPs across a hundred of diverse strains. DNA signatures that discriminate lineages and strain groups, as well as specifically define a particular strain, will be selected. HRM capability to detect and distinguish SNRs will also be evaluated. Screening of a few SNR markers could add resolving power to the genotyping tool under development and might be part of the HRM assays.. Discriminative SNPs will be developed into a molecular typing assay and used to provide further data on the whole ANSES collection. Precise identification of B. anthracis isolates enables epidemiological analyses of natural anthrax cases or forensic investigations of criminal acts as part of the Biotox-Piratox network. Studies will be undertaken (1) to describe the historical, geographic and genetic diversity of the three lineages established in France (A.Br.008/009, A.Br.001/002 and B.Br.CNEVA), (2) to establish the phylogenetic relationships among B. anthracis isolates inside and outside France and (3) to investigate major epizootic recently occurred, such as those in Doubs in 2008 (involving strains affiliated with the A.Br.001/002 sub-group) and in Alps in 2009 (strains allocated into the B.Br.CNEVA sub-lineage). An extensive genotype data set will be created.

Allergenic foods represent a significant health risk for persons who have an allergy to specific food. However, this risk remains poorly characterised. Knowledge is often missing on the different risk components such as the presence of allergens in food, food consumption behaviours of allergic sufferers and thresholds of reaction. Usual risk assessment approaches cannot be easily implemented to assess the risk from sporadic contamination and that concerns only a small part of the population. As a consequence, there is a lack of risk-based guidance and operational tools of those involved in allergen risk management, who are the food industries, the allergic individuals and the food regulators. To upgrade knowledge, methodological and operational tools in food allergy, MIRABEL project involves various scientific fields such as chemical analysis, dietary survey, medical research, socio-economics, applied mathematics and statistics to exposure and risk assessments. In this way, MIRABEL project, which enters in the field of fundamental research, aims to set an integrated and operational framework for the allergic risk analysis, in order to improve quality of life of allergic sufferers. Each component of the risk will be investigated in order to accurately characterize the risk of food allergy and to test different risk management options. To complete existing but sparse information, field surveys will be conducted in order to acquire accurate data on allergic consumers’ behaviours relating to allergen-containing products and their thresholds of reaction, and to allergen presence in food consumed by allergic sufferers. In parallel, methodological developments in Bayesian statistics and probabilistic modelling will be realized to be able to combine the acquired data in an integrated risk quantification model. A cost-benefits analysis for the stakeholders (food industry, allergic individuals and regulators) will also be conducted to anticipate impacts of new policies. The project will be focused on the peanut, which can be adventitiously present in various foodstuffs such as chocolate, cereals or biscuits. This allergen is associated with the highest prevalence of food allergy and is one of the food allergens that lead to the most severe adverse effects.