1. Viral metagenomic analysis of the cheese surface: A comparative study of rapid procedures for extracting viral particles
- Author
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Marie-Agnès Petit, Marianne De Paepe, Stéphane Chaillou, Anne-Sophie Sarthou, Y. Fedala, Eric Dugat-Bony, Julien Lossouarn, Génie et Microbiologie des Procédés Alimentaires (GMPA), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Université Paris Saclay (COmUE), MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL), and Métaprogramme MEM INRA
- Subjects
Viral metagenomics ,viral particles extraction procedure ,Computational biology ,Biology ,Microbiology ,03 medical and health sciences ,chemistry.chemical_compound ,viral metagenomic ,Cheese ,Virology ,Bacteriophages ,Human virome ,Membrane vesicle ,Dna viral ,030304 developmental biology ,2. Zero hunger ,0303 health sciences ,030306 microbiology ,Microbiota ,Virion ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,chemistry ,Metagenomics ,Metagenome ,Chromosomal dna ,Density gradient ultracentrifugation ,DNA ,Food Science ,cheese rind - Abstract
International audience; The structure and functioning of microbial communities from fermented foods, including cheese, have beenextensively studied during the past decade. However, there is still a lack of information about both the occur-rence and the role of viruses in modulating the function of this type of spatially structured and solid ecosystems.Viral metagenomics was recently applied to a wide variety of environmental samples and standardized proce-dures for recovering viral particles from different type of materials has emerged. In this study, we adapted aprocedure originally developed to extract viruses from fecal samples, in order to enable efficient virome analysisof cheese surface. We tested and validated the positive impact of both addition of afiltration step prior to virusconcentration and substitution of purification by density gradient ultracentrifugation by a simple chloroformtreatment to eliminate membrane vesicles. Viral DNA extracted from the several procedures, as well as a vesiclesample, were sequenced using Illumina paired-end MiSeq technology and the subsequent clusters assembledfrom the virome were analyzed to assess those belonging to putative phages, plasmid-derived DNA, or even frombacterial chromosomal DNA. The best procedure was then chosen, and used to describe thefirst cheese surfacevirome, using Epoisses cheese as example. This study provides the basis of future investigations regarding theecological importance of viruses in cheese microbial ecosystems.
- Published
- 2020
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