Meat and seafood products are highly perishable foodstuffs, the preservation of which is crucial. Indeed, a bad preservation of these products may lead to important hygienic and safety risks as well as considerable economic loss, a situation that is particularly true for novel products. The utilisation of protective cultures in order to limit these risks is proposed since several years but is still not in common use, partially because a lack of knowledge of microbial ecosystems of these products. The partners of the French network FLOREPRO decided to share their know-how and scientific knowledge on one hand, and their various expertise about food on the other hand, in order to increase the knowledge of meat and seafood products and on the impact of protective cultures on these ecosystems and on the quality of the products. Our aim is thus to provide a sound scientific based information on the benefit that the utilisation of protective cultures may represent in the production of meat/fish foodstuffs. The project is divided into three tasks: - an as exhaustive as possible characterisation of the ecosystem of height selected products (meat and seafood) by the up to date most performing method, 16S rDNA pyrosequencing of the bacterial DNA present on these products, at the time of production and after the use by date. This task also encompasses the selection of putative spoiler micro-organisms from those products; - set up and sharing of model matrices and sensory analysis in order to reproduce spoilage and to prove the spoiling properties of the strains selected from the first task; - evaluation of the impact of already available protective cultures on spoilage, sensorial quality, and global ecosystem of the products we propose to study. The partners of this project are academics, agro institutes and one SME which was a pioneer in the commercialisation of protective cultures. Other industries, SMEs with no financial support, joined the project to furnish the foodstuffs to be analysed. Know-how, expertise and missions of the partners are complementary and about meat/fish products. The foodstuffs we selected are representative of a large panel of origins (beef, veal, poultry, fish, shellfish) and processes (ground, diced cubes, fillets, peeled) and concern novel products for which knowledge about their ecosystem and sanitary quality is scarce.

Protozoan and Metazoan parasites frequently infest edible fishes worldwide. Some of them are both fish pathogens and recognized agents of important zoonoses with high public health impact. In addition, uncertainty exists about the potential ability of other fish pathogens to infect humans. Meanwhile, parasitized fish can be found often enough in European fish stores. Furthermore, some parasites of fish are able to alter the organoleptic properties of fish products, having therefore a negative impact on fish industry. For these reasons, the present project is targeting fish parasites with impact on both health of consumers and quality of seafood. It proposes a reappraisal of fish-parasite related hazards by developing cost-effective innovative methods to detect eukaryotic pathogens in edible fish, associated with adapted training programs. It focuses mainly on both Anisakidae and Diphyllobothriidae larvae that cause two emerging helminthic diseases: anisakiasis and diphyllobothriasis, respectively. The growing consuming of marinated (or not) raw food, "exotic" or takeaway dishes and marked increase of out-of-home-food could favour the emergence of both typical anisakiasis and Anisakis-induced allergy. Interestingly, 43 % of restaurants and delicatessens did not freeze fish or did not use frozen fish for raw fish dishes. Likewise, the number of human cases of diphyllobothriasis due to the consumption of raw or undercooked freshwater fish from Alpine Lakes increased for the last years. This proposal targeted also the Apicomplexan protists of Cryptosporidium genus. In short, the pathogenic power to humans of fish-derived Cryptosporidium is unknown. We showed recently that C. molnari (from dorada and sea bass) was unable to infect SCID mice. We do not know, however, if other fish-derived Cryptosporidium species or strains are infectious to mammals. The diversity of Cryptosporidium genus could be important in sea- or freshwater teleosteens. We are able to detect, identify and describe new Cryptosporidium species on molecular and phenotypic bases as well as to explore their pathogenic power to mammals in a highly reproducible experimental model well mastered by BDEEP members. The present project is consistent with the "experimental development" kind of research ('Call for proposals', p 24), the integrative online approach proposed here search for "acquiring, associating and using knowledge and scientific, technological, and training methods" in order to generate updated conceptions about edible-fish parasites, and innovative procedures to detect and identify them. Moreover, the present project is suitable for thematic axis 2 of the call for proposals ("Research in alimentation for a more dynamic market of food production") for the following reasons: (a) it proposes a rationale screening activity of parasites harboured by edible fish as an added technological value integrated into an industrial structure or business establishment; (b) it aims at integrating health safety, nutritional quality, organoleptic quality, practical wisdom and reasonable costs; (c) it proposes the combination of know-how of IFREMER specialists in quality of fish products, AFSSA specialists in microbiological health risk and protection of consumers, CEVPM specialists in RD and biotechnology of seafood products, skill of LASMEA and ARBOR Technology in conception and manufacturing of device for food industry; (d) parasitology specialists of three recognized centres: 'Biology & Diversity of Emerging Eukaryotic Pathogens' service of Lille Pasteur Institute (BDEEP, EA3609-Lille-2 University, formerly 'Ecology of Parasitism'), specialized in emerging parasitic Protists and protist-like organisms, Parasitology-Mycology Central Lab of Cochin Hospital (Paris), specialized in Diphyllobothridae tapeworms and Parasitology laboratory of Public Health Sciences, DSSP, Sapienza Roma University (Rome) specialized in metazoan parasites of fish and marine mammals.