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Metagenomic to evaluate norovirus genomic diversity in oysters: Impact on hexamer selection and targeted capture-based enrichment ArchiMer
Strubbia, Sofia; Schaeffer, Julien; Besnard, Alban; Wacrenier, Candice; Le Mennec, Cecile; Garry, Pascal; Desdouits, Marion; Le Guyader, Soizick.
Human virus transmission through food consumption has been identified since many years and the international trade increases the risk of dissemination of viral pathogens. The development of metagenomic approach holds many promises for the surveillance of viruses in food and water. This work aimed to analyze norovirus diversity and to evaluate strain-dependent accumulation patterns in three oyster types by using a metagenomic approach. Different hexamer sets to prime cDNA were evaluated before capture-based approach to enhance virus reads recovery during deep sequencing. The study includes the use of technical replicates of artificially contaminated oysters and the analysis of multiple negatives controls. Results showed a clear impact of the hexamer set...
Tipo: Text Palavras-chave: Norovirus; Oysters; Metagenomic; Human enteric viruses..
Ano: 2020 URL: https://archimer.ifremer.fr/doc/00614/72575/71535.pdf
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Rapport d’activités 2018 – Laboratoire Santé Environnement et Microbiologie. ArchiMer
Desdouits, Marion; Garry, Pascal; Gourmelon, Michele; Hervio Heath, Dominique; Hubert, Francoise; Kaelin, Gaelle; Kergaravat, Cedric; Le Mennec, Cecile; Le Guyader, Soizick; Loiseau, Veronique; Lozach, Solen; Maillot, Jessica; Ollivier, Joanna; Parnaudeau, Sylvain; Piquet, Jean-come; Quenot, Emmanuelle; Rocq, Sophie; Schaeffer, Julien; Serghine, Joelle; Vallade, Emilie; Veron, Antoine; Wacrenier, Candice.
Ce rapport présente une synthèse des actions, et travaux réalisés par le laboratoire Santé Environnement et Microbiologie pendant l’année 2018. Les divers projets de recherche sont résumés et les derniers développements ou résultats sont brièvement évoqués. Les actions en tant que Laboratoire National de Référence pour la Microbiologie des Coquillages et les activités pour la coordination du réseau REMI sont également présentées.
Tipo: Text Palavras-chave: Microbiologie sanitaire; Bactéries entériques; Vibrions; Virus entériques humains; Norovirus; Coquillage; Activité de référence; REMI..
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00491/60218/63584.pdf
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A Targeted Metagenomics Approach to Study the Diversity of Norovirus GII in Shellfish Implicated in Outbreaks ArchiMer
Desdouits, Marion; Wacrenier, Candice; Ollivier, Joanna; Schaeffer, Julien; Le Guyader, Soizick.
Human noroviruses (NoV) cause epidemics of acute gastroenteritis (AGE) worldwide and can be transmitted through consumption of contaminated foods. Fresh products such as shellfish can be contaminated by human sewage during production, which results in the presence of multiple virus strains, at very low concentrations. Here, we tested a targeted metagenomics approach by deep-sequencing PCR amplicons of the capsid (VP1) and polymerase (RdRp) viral genes, on a set of artificial samples and on shellfish samples associated to AGE outbreaks, to evaluate its advantages and limitations in the identification of strains from the NoV genogroup (G) II. Using artificial samples, the method allowed the sequencing of most strains, but not all, and displayed variability...
Tipo: Text Palavras-chave: Norovirus; Foodborne virus; Metagenomics; Amplicon deep sequencing; Viral diversity.
Ano: 2020 URL: https://archimer.ifremer.fr/doc/00646/75831/76818.pdf
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Passive Samplers, a Powerful Tool to Detect Viruses and Bacteria in Marine Coastal Areas ArchiMer
Vincent Hubert, Francoise; Wacrenier, Candice; Morga, Benjamin; Lozach, Solen; Quenot, Emmanuelle; Mege, Mickael; Lecadet, Cyrielle; Gourmelon, Michele; Hervio-heath, Dominique; Le Guyader, Soizick.
The detection of viruses and bacteria which can pose a threat either to shellfish health or shellfish consumers remains difficult. The current detection methods rely on point sampling of water, a method that gives a snapshot of the microorganisms present at the time of sampling. In order to obtain better representativeness of the presence of these microorganisms over time, we have developed passive sampling using the adsorption capacities of polymer membranes. Our objectives here were to assess the feasibility of this methodology for field detection. Different types of membrane were deployed in coastal waters over 2 years and the microorganisms tested using qPCR were: human norovirus (NoV) genogroups (G)I and II, sapovirus, Vibrio spp. and the species...
Tipo: Text Palavras-chave: Norovirus; Ostreid herpes virus 1 μ Var; Vibrio spp; Microbial source tracking; Sea; Passive sampler; Oyster (Crassostrea gigas).
Ano: 2021 URL: https://archimer.ifremer.fr/doc/00682/79375/81893.pdf
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Metavirome Sequencing to Evaluate Norovirus Diversity in Sewage and Related Bioaccumulated Oysters ArchiMer
Strubbia, Sofia; Schaeffer, Julien; Oude Munnink, Bas B.; Besnard, Alban; Phan, My V. T.; Nieuwenhuijse, David F.; De Graaf, Miranda; Schapendonk, Claudia M. E.; Wacrenier, Candice; Cotten, Matthew; Koopmans, Marion P. G.; Le Guyader, Soizick.
Metagenomic sequencing is a promising method to determine the virus diversity in environmental samples such as sewage or shellfish. However, to identify the short RNA genomes of human enteric viruses among the large diversity of nucleic acids present in such complex matrices, method optimization is still needed. This work presents methodological developments focused on norovirus, a small ssRNA non-enveloped virus known as the major cause of human gastroenteritis worldwide and frequently present in human excreta and sewage. Different elution protocols were applied and Illumina MiSeq technology were used to study norovirus diversity. A double approach, agnostic deep sequencing and a capture-based approach (VirCapSeq-VERT) was used to identify norovirus in...
Tipo: Text Palavras-chave: Norovirus; Sewage; Oysters; Metagenomic sequencing; Metavirome.
Ano: 2019 URL: https://archimer.ifremer.fr/doc/00586/69765/67656.pdf
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PERLE - Programme d'expérimentation et de recherche sur l'huître plate Ostrea edulis - Rapport final des sous-programmes 2 et 3 ArchiMer
Arzul, Isabelle; Baillon, Laury; Boudry, Pierre; Bouget, Jean-francois; Chollet, Bruno; Cochet, Helene; Couraleau, Yann; Dubreuil, Christine; Duchene, Julie; Dutartre, Julie; Garcia, Celine; Heloury, Purotu; Heurtebise, Serge; Huchet, Eve; Hussenot, Mathieu; Langlade, Aime; Ollier, Simon; Omnes, Emmanuelle; Pouvreau, Stephane; Roby, Charlotte; Serpin, Delphine; Viard, Frederique; Wacrenier, Candice; Lapegue, Sylvie.
L’huître plate (Ostrea edulis) est une espèce endémique des côtes européennes, elle est présente depuis des millions d’années. Son aire de répartition est très vaste allant de la Norvège jusqu’au Maroc. Elle est également présente en mer Méditerranée, en mer Noire, mais également au Canada, aux Etats-Unis ou encore en Afrique du Sud, à la suite d'introductions pour des raisons aquacoles. En France, à partir de la fin des années 70, de fortes mortalités ont été constatées sur les bancs naturels et en élevage, faisant rapidement chuter la production de 20 000 tonnes à 2 000 tonnes. Ces mortalités ont été attribuées à deux maladies parasitaires : la marteiliose (due au parasite Marteilia refringens) et la bonamiose (due au parasite Bonamia ostreae). Trente...
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Ano: 2014 URL: http://archimer.ifremer.fr/doc/00247/35815/34328.pdf
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