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1. Fate and impact of nanoplastics in the human digestive environment after oral exposure: A common challenge for toxicology and chemistry

Author

Liebgott, Chloé, Chaib, Iseline, Doyen, Périne, Robert, Hervé, Eutamene, Hélène, Duflos, Guillaume, Reynaud, Stéphanie, Grassl, Bruno, Mercier-Bonin, Muriel, Laboratoire de Boulogne sur mer, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Biochimie des Produits Aquatiques (BPA), Université du Littoral Côte d'Opale (ULCO)-Transfrontalière BioEcoAgro - UMR 1158 (BioEcoAgro), Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-JUNIA (JUNIA), Université catholique de Lille (UCL)-Université catholique de Lille (UCL)-Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-JUNIA (JUNIA), Université catholique de Lille (UCL)-Université catholique de Lille (UCL), Transfrontalière BioEcoAgro - UMR 1158 (BioEcoAgro), INRAe, Anses, Région Hauts-de-France, and Thèse ScreenPlastiFood

Subjects
Gut toxicity, Chemistry, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, In vitro and in vivo models, Human health, Nanoplastics, Toxicology, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, NPL model materials
Abstract

International audience; Nanoplastics (NPLs), the presence in the environment of which was considered only “highly plausible” until recently, have become the focus of environmental and ecotoxicological studies. However, up to know, little is known about the potential risks of NPLs to human health. In this review, we provide an overview of the evidence of a specific impact of NPLs on human digestive health reported to date. We focus on the different sources of oral exposure to NPLs, including food packaging, food and beverages. We then summarize the toxicological effects of such exposure on the digestive ecosystem in vitro and in vivo in rodents. Importantly, gut toxicity results should be assessed with the type of NPL model materials employed, due to the critical influence of their chemical and physical properties. As a result, we thoroughly describe NPLs with their source, chemical composition and physicochemical behavior to emphasize the lack of NPL characterization and/or model materials. Finally, we propose avenues for interdisciplinary studies at the interface of toxicology and chemistry, with a view to achieving appropriate scientific assessments of the risks to gut health posed by NPLs, and improvements in their management.

Published
2023
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2. Répartition des microplastiques dans la colonne d'eau d'une rivière lors de différentes conditions environnementales

Author

Pasquier, Gabriel, Doyen, Périne, Dehaut, Alexandre, Veillet, Guillaume, Duflos, Guillaume, Amara, Rachid, Laboratoire d’Océanologie et de Géosciences (LOG) - UMR 8187 (LOG), Institut national des sciences de l'Univers (INSU - CNRS)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Nord]), Université du Littoral Côte d'Opale (ULCO), Biochimie des Produits Aquatiques (BPA), Université du Littoral Côte d'Opale (ULCO)-Transfrontalière BioEcoAgro - UMR 1158 (BioEcoAgro), Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-JUNIA (JUNIA), Université catholique de Lille (UCL)-Université catholique de Lille (UCL)-Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-JUNIA (JUNIA), Université catholique de Lille (UCL)-Université catholique de Lille (UCL), Laboratoire de Boulogne sur mer, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), and GDR Polymères et Océans

Subjects
[CHIM.ANAL]Chemical Sciences/Analytical chemistry, [SDE]Environmental Sciences
Abstract

International audience; Dans le cadre de cette étude, la contamination en microplastiques a été évaluée dans une rivière à la fois en surface (0-25cm) et en subsurface (25-50cm). La partie la plus profonde de la colonne d’eau (10 derniers cm) a également été échantillonnée ainsi les sédiments. Trois campagnes d’échantillonnage ont été menées lors de différentes conditions météorologiques : durant une forte pluie au printemps, après une forte pluie en automne et lors d’une période sèche en hiver. Les résultats montrent que les eaux de subsurface contiennent autant de particules que les eaux de surface, quelles que soient les conditions environnementales.Des concentrations en particules 3 à 11 fois supérieures ont été retrouvées lors des fortes pluies au printemps comparées aux autres campagnes d’échantillonnage dans tous les compartiments étudiés dans la colonne d’eau. Tous compartiments confondus, le type de particules observé est impacté par les conditions météorologiques avec 86% à 96% de fibres parmi les particules échantillonnées en hiver, comparé aux autres campagnes au printemps et en automne où les fibres représentent respectivement 31% et 75%.Lors de cette étude 73,5% des particules analysées sont des polymères qui ont une densité théorique inférieure à 1 dans les échantillons de surface et subsurface, alors que cette proportion tombe à 40,8% pour les échantillons de fond et de sédiments ce qui indique une répartition des particules selon leur densité dans la colonne d’eau.Cette étude souligne l’importance de prendre en compte la subsurface ainsi que les variations saisonnières pour évaluer les types et niveaux de contamination microplastiques dans les environnements fluviaux.

Published
2023

3. Translocation de nanoplastiques à travers la barrière intestinale du bar commun (Dicentrarchus labrax) : preuve expérimentale à l’aide de nanobilles de polystyrène modèle

Author

Dehaut, Alexandre, Vagner, Marie, Boudry, Gaëlle, Courcot, Lucie, Vincent, Dorothée, Duflos, Guillaume, Huvet, Arnaud, Tallec, Kévin, Zambonino-Infante, Jose-Luis, Laboratoire de Boulogne sur mer, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Nutrition, Métabolismes et Cancer (NuMeCan), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire d’Océanologie et de Géosciences (LOG) - UMR 8187 (LOG), Institut national des sciences de l'Univers (INSU - CNRS)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Nord]), Université du Littoral Côte d'Opale (ULCO), Office français de la biodiversité (OFB), CPER MARCO 2015 - 2020, Société Française d'Ichtyologie, and ANR-15-CE34-0006,Nanoplastics,Microplastiques, nanoplastiques dans l'environnement marin: caractérisation, impacts et évaluation des risques sanitaires.(2015)

Subjects
billes, MEB - Microscopie électronique à balayage, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Nanoplastiques, Dicentrachus labrax, PS, Fluorimétrie, [SDV.TOX.ECO]Life Sciences [q-bio]/Toxicology/Ecotoxicology, Bar commun, Py-GC-MS
Abstract

National audience; La pollution plastique de tous les compartiments de l’environnement, y compris l’environnement marin, n’est désormais plus à démontrer. Concomitamment à cette pollution, la présence de particules plastiques de tailles micrométriques et nanométriques a également été rapportée. Ces particules peuvent constituer un danger physique ou agir en tant que vecteurs de différents contaminants chimiques ou biologiques. Chez les organismes vivants le devenir de ces particules n’est que partiellement connu et le passage des barrières biologiques reste controversé, bien qu’il soit probable pour les particules les plus petites comme les nanoplastiques. Afin de répondre à cette question de la possible translocation de nanoplastiques, des manipulations d’exposition ex vivo à des nanobilles de polystyrène modèle d’une taille de 50 nm (NP-PS) ont été réalisées sur des intestins de bar commun (Dicentrarchus labrax) en utilisant des chambres de Ussing.Grâce à la combinaison des résultats obtenus par plusieurs techniques de détection : fluorométrie, microscopie électronique à balayage (MEB) et pyrolyse couplée à la chromatographie en phase gazeuse et à un spectromètre de masse haute-résolution (Py-GC-HRMS), il a été possible d’enregistrer une augmentation de la fluorescence du côté séreux quelques dizaines de minutes après l’exposition des échantillons à la solution de NP-PS. Cette mesure a pu être confirmée par celles réalisées en MEB et Py-GC-HRMS. La translocation de nanobilles de PS de 50nm et sans agrégation dans le milieu expérimental, à travers la paroi de l’intestin de cette espèce de poisson a donc été attestée.

Published
2023

4. Oral exposure to polyethylene microplastics alters gut morphology, immune response, and microbiota composition in mice

Author

Madjid Djouina, Cécile Vignal, Alexandre Dehaut, Ségolène Caboche, Nell Hirt, Christophe Waxin, Charlotte Himber, Delphine Beury, David Hot, Laurent Dubuquoy, David Launay, Guillaume Duflos, Mathilde Body-Malapel, Institute for Translational Research in Inflammation - U 1286 (INFINITE (Ex-Liric)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Plateformes Lilloises en Biologie et Santé - UAR 2014 - US 41 (PLBS), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), and Laboratoire de Boulogne sur mer

Subjects
Inflammation, [SDV]Life Sciences [q-bio], Microbiota, Microplastics, Immunity, Biochemistry, Mice, Intestinal, Polyethylene, MESH: Microbiota, Animals, MESH: Microplastics, Plastics, ComputingMilieux_MISCELLANEOUS, Biomarkers, Water Pollutants, Chemical, General Environmental Science
Abstract

The ubiquitous and growing presence of microplastics (MPs) in all compartments of the environment raises concerns about their possible harmful effects on human health. Human exposure to MPs occurs largely through ingestion. Polyethylene (PE) is widely employed for reusable bags and food packaging and found to be present in drinking water and food. It is also one of the major polymers detected in human stool. The aim of this study was to characterize the effects of intestinal exposure to PE MPs on gut homeostasis. Mice were orally exposed for 6 weeks to PE microbeads of 2 different sizes, 36 and 116 μm, that correspond to those found in human stool. They were administrated either individually or as a mixture at a dose of 100 μg/g of food. Both PE microbead sizes were detected in mouse stool. Different parameters related to major intestinal functions were compared between control mice, mice exposed to each type of microbead, or co-exposed to the 2 types of microbeads. Intestinal disturbances were observed after individual exposure to each size of PE microbead, and the most marked deleterious effects were found in co-exposed mice. At the histomorphological level, crypt depth was increased throughout the intestinal tissues. Significant variations of gene expression related to epithelial, permeability, and inflammatory biomarkers were quantified. Defective recruitment of some intestinal immune cells was observed from the proximal portion of the small intestine to the colon. Several bacterial taxa at the order level were found to be affected by exposure to the MPs by metagenomic analysis of cecal microbiota. These results show that ingestion of PE microbeads induces significant alterations of crucial intestinal markers in mice and underscores the need to further study the health impact of MP exposure in humans.

Published
2022
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5. Identification and quantification of plastic additives using Pyrolysis-GC/MS: a review

Author

Chaza Chbib, Sébastien Monchy, Alexandre Dehaut, Périne Doyen, Guillaume Duflos, Ika Paul-Pont, Fleurine Akoueson, Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Université du Littoral Côte d'Opale (ULCO), Laboratoire d’Océanologie et de Géosciences (LOG) - UMR 8187 (LOG), Centre National de la Recherche Scientifique (CNRS)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut national des sciences de l'Univers (INSU - CNRS), Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut Charles Viollette (ICV) - ULR 7394 (ICV), Université d'Artois (UA)-Université du Littoral Côte d'Opale (ULCO)-Institut Supérieur d'Agriculture-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Région Hauts-de-France, Anses, CPER Marco (Union Européenne FEDER, Etat Français, Région Hauts-de-France, Ifremer), Thèse Addimicro, CPER Marco (2015-2020), Laboratoire de Boulogne sur mer, Biochimie des Produits Aquatiques (BPA), Université du Littoral Côte d'Opale (ULCO)-Transfrontalière BioEcoAgro - UMR 1158 (BioEcoAgro), Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-JUNIA (JUNIA), Université catholique de Lille (UCL)-Université catholique de Lille (UCL)-Université d'Artois (UA)-Université de Liège-Université de Picardie Jules Verne (UPJV)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-JUNIA (JUNIA), Université catholique de Lille (UCL)-Université catholique de Lille (UCL), Transfrontalière BioEcoAgro - UMR 1158 (BioEcoAgro), Institut national des sciences de l'Univers (INSU - CNRS)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Nord]), and Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Environmental Engineering, Materials science, 010504 meteorology & atmospheric sciences, 010501 environmental sciences, 01 natural sciences, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Environmental Chemistry, Sample preparation, Gas chromatography mass spectrometer, multi-shot pyrolysis, Process engineering, Waste Management and Disposal, 0105 earth and related environmental sciences, business.industry, Pyrolysis gc ms, Polymeric matrix, Limiting, Py-GC/MS, Pollution, Plastic polymers, Identification (information), Organic plastic additives (OPAs), [CHIM.POLY]Chemical Sciences/Polymers, plastic polymers, Multi-shot pyrolysis, business
Abstract

International audience; Analysis of organic plastic additives (OPAs) associated to plastic polymers is growing. The current review outlines the characteristics and the development of (multi-step) pyrolysis coupled with a gas chromatography mass spectrometer (Py-GC/MS) for the identification and semi-quantification of OPAs. Compared to traditional methods, Py-GC/MS offers advantages like suppressing extensive steps of preparation, limiting contamination due to solvents and the possibility to analyse minute particles. Its key advantage is the successive analysis of OPAs and the polymeric matrix of the same sample. Based on the studied articles, numerous methods have been described allowing identification and, in some case, semi-quantification of OPAs. There is nevertheless no gold standard method, especially given the huge diversity of OPAs and the risks of interferences with polymers or other additives, but, among other parameters, a consensus temperature seems to arise from studies. More broadly, this review also explores many aspects on the sample preparation like weight and size of particles and calibration strategies. After studying the various works, some development prospects emerge and it appears that methodological developments should focus on better characterizing the limits of the methods in order to consider which OPAs can be quantified and in which polymers this is feasible.

Published
2021
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6. Dynamics of mobile genetic elements of Listeria monocytogenes persisting in ready-to-eat seafood processing plants in France

Author

Ludovic Mallet, Graziella Midelet-Bourdin, Federica Palma, Thomas Brauge, Anne Brisabois, Nicolas Radomski, Michel-Yves Mistou, Arnaud Felten, Laurent Guillier, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Mathématiques et Informatique Appliquées du Génome à l'Environnement [Jouy-En-Josas] (MaIAGE), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), European Project: 643476,H2020,H2020-PHC-2014-single-stage,COMPARE(2014), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), and Laboratoire de Boulogne sur mer

Subjects
Food processing plant, Prophage profiling, Prophages, [SDV]Life Sciences [q-bio], medicine.disease_cause, MESH: Listeria monocytogenes, Persistent clones, Plasmid, Genomic island, Phylogeny, 2. Zero hunger, Genetics, 0303 health sciences, Genomics, Horizontal gene transfer, France, Plasmids, Research Article, Biotechnology, lcsh:QH426-470, Genomic Islands, Horizontal plasmid transfer, lcsh:Biotechnology, Biology, Polymorphism, Single Nucleotide, 03 medical and health sciences, Listeria monocytogenes, Stress, Physiological, lcsh:TP248.13-248.65, medicine, Food-Processing Industry, Prophage, 030304 developmental biology, Comparative genomics, 030306 microbiology, biology.organism_classification, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Interspersed Repetitive Sequences, lcsh:Genetics, Seafood, Genes, Bacterial, Mobile genetic elements, Listeria, Genome, Bacterial
Abstract

BackgroundListeria monocytogenesClonal Complexes (CCs) have been epidemiologically associated with foods, especially ready-to-eat (RTE) products for which the most likely source of contamination depends on the occurrence of persisting clones in food-processing environments (FPEs). As the ability ofL. monocytogenesto adapt to environmental stressors met in the food chain challenges the efforts to its eradication from FPEs, the threat of persistent strains to the food industry and public health authorities continues to rise. In this study, 94 food and FPEsL. monocytogenesisolates, representing persistent subtypes contaminating three French seafood facilities over 2–6 years, were whole-genome sequenced to characterize their genetic diversity and determine the biomarkers associated with long-term survival in FPEs.ResultsFood and FPEs isolates belonged to five CCs, comprising long-term intra- and inter-plant persisting clones. Mobile genetic elements (MGEs) such as plasmids, prophages and transposons were highly conserved within CCs, some of which harboured genes for resistance to chemical compounds and biocides used in the processing plants. Some of these genes were found in a 90.8 kbp plasmid, predicted to be” mobilizable”, identical in isolates from CC204 and CC155, and highly similar to an 81.6 kbp plasmid from isolates belonging to CC7. These similarities suggest horizontal transfer between isolates, accompanied by deletion and homologous recombination in isolates from CC7. Prophage profiles characterized persistent clonal strains and several prophage-loci were plant-associated. Notably, a persistent clone from CC101 harboured a novel 31.5 kbp genomic island that we namedListeriagenomic island 3 (LGI3), composed by plant-associated loci and chromosomally integrating cadmium-resistance determinantscadA1C.ConclusionsGenome-wide analysis indicated that inter- and intra-plant persisting clones harbour conserved MGEs, likely acquired in FPEs and maintained by selective pressures. The presence of closely related plasmids inL. monocytogenesCCs supports the hypothesis of horizontal gene transfer conferring enhanced survival to FPE-associated stressors, especially in hard-to-clean harbourage sites. Investigating the MGEs evolutionary and transmission dynamics provides additional resolution to trace-back potentially persistent clones. The biomarkers herein discovered provide new tools for better designing effective strategies for the removal or reduction of residentL. monocytogenesin FPEs to prevent contamination of RTE seafood.

Published
2020
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7. First Report on the Prevalence and Subtype Distribution of Blastocystis sp. in Edible Marine Fish and Marine Mammals: A Large Scale-Study Conducted in Atlantic Northeast and on the Coasts of Northern France

Author

Nausicaa Gantois, Angélique Lamot, Yuwalee Seesao, Colette Creusy, Luen-Luen Li, Sébastien Monchy, Sadia Benamrouz-Vanneste, Jacky Karpouzopoulos, Jean-Luc Bourgain, Célia Rault, Fabien Demaret, Martha Baydoun, Magali Chabé, Emilie Fréalle, Cécile-Marie Aliouat-Denis, Mélanie Gay, Gabriela Certad, Eric Viscogliosi, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Boulogne sur mer, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Groupement des Hôpitaux de l'Institut Catholique de Lille (GHICL), Laboratoire d’Océanologie et de Géosciences (LOG) - UMR 8187 (LOG), Centre National de la Recherche Scientifique (CNRS)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Institut national des sciences de l'Univers (INSU - CNRS), Institut Catholique de Lille (ICL), Université catholique de Lille (UCL), Groupe Mammifères Marins, Coordination Mammalogique du Nord de la France [Vimy] (CMNF), Observatoire PELAGIS UMS 3462 (PELAGIS), LIttoral ENvironnement et Sociétés - UMRi 7266 (LIENSs), Université de La Rochelle (ULR)-Centre National de la Recherche Scientifique (CNRS)-Université de La Rochelle (ULR)-Centre National de la Recherche Scientifique (CNRS), This study was supported by the Conseil Régional Hauts-de-France (Concerted Research Actions of Regional Initiative ARCir 13 ABC FISH No. 13003283), the regional competitiveness centre AQUIMER (Boulogne-sur-Mer, France), the Programme Orientations Stratégiques of the University of Lille, the Centre National de la Recherche Scientifique, the Institut Pasteur of Lille, the University Catholic of Lille, the French Agency for Food, Environmental and Occupational Health and Safety, the French Agency for Biodiversity and the Agglomeration Community of the City of La Rochelle. Y.S. was supported by a PhD fellowship from the Ministry of Science and Technology of Thailand (001/2555)., Institut national des sciences de l'Univers (INSU - CNRS)-Université du Littoral Côte d'Opale (ULCO)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD [France-Nord]), Université du Littoral Côte d'Opale (ULCO), Observatoire pour la Conservation de la Mégafaune Marine (PELAGIS), LIttoral ENvironnement et Sociétés (LIENSs), and La Rochelle Université (ULR)-Centre National de la Recherche Scientifique (CNRS)-La Rochelle Université (ULR)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Microbiology (medical), subtyping, 030231 tropical medicine, Zoology, Distribution (economics), Biology, intestinal parasites, Microbiology, [SDV.MP.PRO]Life Sciences [q-bio]/Microbiology and Parasitology/Protistology, molecular epidemiology, Article, law.invention, 03 medical and health sciences, 0302 clinical medicine, law, Virology, medicine, Parasite hosting, 14. Life underwater, marine mammals, lcsh:QH301-705.5, Polymerase chain reaction, edible marine fish, Genetic diversity, Blastocystis, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, Molecular epidemiology, business.industry, Zoonosis, transmission, Blastocystis sp, zoonosis, medicine.disease, biology.organism_classification, ssu rdna sequence, 030104 developmental biology, Transmission (mechanics), lcsh:Biology (General), blastocystis sp, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, real-time quantitative pcr, business
Abstract

International audience; Blastocystis is frequently identified in humans and animal hosts and exhibits a large genetic diversity with the identification of 17 subtypes (STs). Despite its zoonotic potential, its prevalence and ST distribution in edible marine fish and marine mammals remain unknown. A large-scale survey was thus conducted by screening 345 fish caught in Atlantic Northeast and 29 marine mammals stranded on the coasts of northern France for the presence of the parasite using real-time Polymerase Chain Reaction PCR. The prevalence of the parasite was about 3.5% in marine fish. These animals were mostly colonized by poikilotherm-derived isolates not identified in humans and corresponding to potential new STs, indicating that fish are natural hosts of Blastocystis. Marine fishes are also carriers of human STs and represent a likely limited source of zoonotic transmission. 13.8% of the marine mammals tested were colonized and 6 different STs were identified including 3 potential new STs. The risk of zoonotic transmission through marine mammals is insignificant due to the lack of repeated contact with humans. The present survey represents the first data regarding the prevalence and ST distribution of Blastocystis in marine fish and marine mammals and provides new insights into its genetic diversity, host range and transmission.

Published
2020
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8. Treatment with disinfectants may induce an increase in viable but non culturable populations of Listeria monocytogenes in biofilms formed in smoked salmon processing environments

Author

Guylaine Leleu, Aurélie Hanin, Graziella Midelet, Catherine Denis, Christine Faille, Thomas Brauge, Laboratoire de Boulogne sur mer, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Unité Matériaux et Transformations - UMR 8207 (UMET), Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Centrale Lille Institut (CLIL), ACTALIA [Villers-Bocage], FranceAgriMer Research Project VINOCU, AQUIMER, FEAMP as part of the SILVERPROTECT project, Centrale Lille-Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and Actalia [Saint-Lô]

Subjects
Microbiological culture, Food Handling, Food Contamination, medicine.disease_cause, Microbiology, [SPI.MAT]Engineering Sciences [physics]/Materials, VBNC, 03 medical and health sciences, chemistry.chemical_compound, Contact surfaces, food, Listeria monocytogenes, Salmon, Propidium monoazide, Fish Products, medicine, Animals, Ammonium, Food science, Hydrogen peroxide, 030304 developmental biology, 0303 health sciences, Microbial Viability, 030306 microbiology, Biofilm, PMA-qPCR, Food environment industry, Hydrogen Peroxide, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, food.food, Quaternary Ammonium Compounds, Smoked salmon, Disinfection, chemistry, 13. Climate action, Biofilms, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Disinfectants, Food Science
Abstract

International audience; The objectives of this study were 1) to evaluate the impact of two industrial disinfectants on the viability ofListeria monocytogenespopulations in biofilm and 2) to investigate the viability state ofL. monocytogenescellspresent on contact surfaces in the smoked salmon processing environment. In thefirst step, we cultured monospecies and mixed species biofilms containingL. monocytogeneson stainless steel or polyvinyl chloride (PVC) at8 °C for 48h. The biofilms were then exposed to quaternary ammonium- and hydrogen peroxide-based disin-fectants. Residual total populations ofL. monocytogeneswere measured by qPCR, and viable culturable (VC) cellpopulations were quantified using standard microbiological culture-based techniques and by a quantitative PCR(qPCR) assay coupled with a propidium monoazide treatment. Decreases in VC populations and the appearanceof viable but non culturable (VBNC) populations were observed in response to treatment with the disinfectants.An 8 month sampling campaign in 4 smoked salmon processing plants was also carried out to detectL. mono-cytogenesin environmental samples. VBNC cells were detected mainly after the cleaning and disinfection op-erations. This study showed that industrial disinfectants did not inactivate allL. monocytogenescells on inertsurfaces. The presence of VBNC populations ofL. monocytogenesin the smoked salmon processing environment isa public health concern.1.

Published
2020
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9. Comparison of the performance of the biofilm sampling methods (swab, sponge, contact agar) in the recovery of Listeria monocytogenes populations considering the seafood environment conditions

Author

Thomas Brauge, Catherine Denis, Christine Faille, Guylaine Leleu, Graziella Midelet, Aurélie Hanin, Unité Matériaux et Transformations - UMR 8207 (UMET), Centrale Lille-Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de Boulogne sur mer, ACTALIA [Villers-Bocage], Actalia [Saint-Lô], French FranceAgriMer Research Project VINOCUP, Hauts-de-France region for funding the Interreg Veg-I-Tec project (programme Interreg V France-Wallonia-Flanders, GoToS3NuTeX), AQUIMER, and Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Centrale Lille Institut (CLIL)

Subjects
Azides, food.ingredient, Disinfectant, Carnobacterium, medicine.disease_cause, Pseudomonas fluorescens, Microbiology, [SPI.MAT]Engineering Sciences [physics]/Materials, 03 medical and health sciences, food, Listeria monocytogenes, medicine, Agar, Neutralizing buffer, Animals, 14. Life underwater, Food science, 030304 developmental biology, 0303 health sciences, biology, Sampling methods VBNC, 030306 microbiology, Biofilm, Fishes, General Medicine, biology.organism_classification, Stainless Steel, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, food.food, Smoked salmon, Europe, Sponge, Seafood, Underestimation, Biofilms, Food Microbiology, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, Bacteria, Food Science, Disinfectants, Propidium
Abstract

Aims The aim of this study was to evaluate the performance of sampling methods [contact plates, sponges, and swabs] in the recovery of biofilm Listeria monocytogenes populations considering the seafood environment conditions (nature of conditioning, of materials and bacterial species). Methods and results Different materials (stainless steel, polyvinyl chloride, polyurethane) were conditioned with two fish filtrates, the ready-to-eat the most consumed in Europe (smoked salmon, cod). After, we added the suspension of Listeria monocytogenes, alone or with Pseudomonas fluorescens or Carnobacterium strains, and incubated for 48 h at 8 °C. Then, the 48 h-biofilms were sampled with different methods (contact plates, sponges, and swabs). The cultivable bacterial populations were enumerated on agar, while the L. monocytogenes total and viable populations were quantified by qPCR and propidium monoazide-qPCR (PMA-qPCR), respectively. The amount of L. monocytogenes in biofilms was affected only by the nature of the conditioning with lowest adherent bacteria with cod versus with smoked salmon conditioning. Considering the amount of total population, the swab displayed the lowest values versus the sponges and the contact plates. An explanation was that the observations of the swab by microscopy showed the bacteria trapped within it. The recovery of cultivable bacterial populations was not significantly different with the three sampling methods. On the contrary, we showed that the VBNC populations were only detached by two of three methods (contact plates, sponges) while for the dead populations, those were contact plates and swabs. Conclusions The nature of the conditioning influenced the amount of the bacteria in biofilms. And the performance of the recovery of the bacterial populations (dead, VBNC, cultivable) was dependent on the methods used. Significance and impact of the study This study showed that the seafood environmental conditions influenced the biofilm formation and the assessment of the efficiency of cleaning and disinfectant operations could be significantly affected by the used sampling methods.

Published
2019
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10. An Irgafos® 168 story: When the ubiquity of an additive prevents studying its leaching from plastics

Author

Arnaud Huvet, Alexandre Dehaut, Charlotte Himber, Justine Receveur, Ronan Jezequel, Ludovic Hermabessiere, Ika Paul-Pont, Philippe Soudant, Christophe Lambert, Fabienne Lagarde, Guillaume Duflos, David Mazurais, Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Laboratoire de Boulogne sur mer, Centre de documentation de recherche et d'expérimentations sur les pollutions accidentelles des eaux (Cedre), Cedre, Laboratoire des Sciences de l'Environnement Marin (LEMAR) (LEMAR), Institut de Recherche pour le Développement (IRD)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER)-Université de Brest (UBO)-Institut Universitaire Européen de la Mer (IUEM), Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Institut Français de Recherche pour l'Exploitation de la Mer - Brest (IFREMER Centre de Bretagne), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Institut des Molécules et Matériaux du Mans (IMMM), Le Mans Université (UM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Anses, Région de Hauts-de-France, CPER MARCO 2015 - 2020, CPER MARCO 2015-2020, ANR-15-CE34-0006,Nanoplastics,Microplastiques, nanoplastiques dans l'environnement marin: caractérisation, impacts et évaluation des risques sanitaires.(2015), and Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Brest (UBO)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Microplastics, Environmental Engineering, 010504 meteorology & atmospheric sciences, 010501 environmental sciences, 01 natural sciences, chemistry.chemical_compound, Irgafos 168 (R), Contamination, [CHIM.ANAL]Chemical Sciences/Analytical chemistry, Environmental Chemistry, Additive, MESH: Microplastics, Waste Management and Disposal, 0105 earth and related environmental sciences, [SDV.EE]Life Sciences [q-bio]/Ecology, environment, Microplastic, Polyethylene, Phosphate, Pollution, 4-di-tert-butylphenyl)phosphite, Irgafos 168®, chemistry, 13. Climate action, [SDV.TOX]Life Sciences [q-bio]/Toxicology, Reagent, Environmental chemistry, Leaching, Leaching (metallurgy), [SDE.BE]Environmental Sciences/Biodiversity and Ecology, MESH: Pyrolysis, Plastic pollution, Tris(2, Pyrolysis
Abstract

International audience; Plastic pollution is a source of chemical to the environment and wildlife. Despite the ubiquity of plastic pollution and thus plastic additive in the environment, plastic additives have been studied to a limited extend. As a prerequisite to a study aiming to evaluate the leaching of a common additive used as an antioxidant (Irgafos® 168) from polyethylene microparticles, an inventory of the potential background contamination of the laboratory workplace was done. In this study, Irgafos® 168 (tris(2,4-ditert-butylphenyl) phosphite) and its oxidized form (tris (2,4-ditert-butylphenyl) phosphate) were quantified in different laboratory reagents, including the plastic packaging and the powders, using Pyrolysis-GC/MS. At least one form of Irgafos® 168 was detected in all tested laboratory reagents with higher concentrations in caps and bottles as compared to the powders. Additionally, oxidized Irgafos® 168 was also found in the reverse osmosed and deionized water container used in the laboratory. The same profile of contamination, i.e. higher concentration of the oxidized form and higher concentrations in acidic reagents, was observed when comparing the reagent and their respective containers suggesting that the additive is leaching from the container into the powder. Overall, this study demonstrates that the antioxidant additive Irgafos® 168 is ubiquitous in the laboratory workplace. Plastic additives such as Irgafos® 168 can therefore largely interfere and biased ecotoxicological and toxicological studies especially using environmentally relevant concentrations of microplastics. The source, fate and effects of plastic additive from plastic debris should be carefully considered in future studies that require setting up methods to overcome these contaminations

Published
2020
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11. Enterococcus faecalis Countermeasures Defeat a Virulent Picovirinae Bacteriophage

Author

Astrid Bouteau, Olivier Son, François Lecointe, Pascale Serror, Julien Lossouarn, Elisabeth Moncaut, Michael S. DuBow, Marie-Agnès Petit, Arnaud Briet, Magali Leroy, Sylviane Furlan, MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Université Paris-Saclay, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Laboratoire de microbiologie des environnements extrêmophiles (LM2E), Centre National de la Recherche Scientifique (CNRS)-Université de Brest (UBO)-Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Anses Laboratoire de Boulogne sur Mer, Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, Université Paris-Sud - Paris 11 (UP11), Génomique et Biodiversité microbienne des biofilms (LGBMB), Département Microbiologie (Dpt Microbio), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Institut de Biologie Intégrative de la Cellule (I2BC), Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay-Université Paris-Sud - Paris 11 (UP11)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Saclay, and Lossouarn, Julien

Subjects
0301 basic medicine, family, prophage, medicine.drug_class, [SDV]Life Sciences [q-bio], 030106 microbiology, Antibiotics, lcsh:QR1-502, Virulence, system, phage abortive infection, lcsh:Microbiology, Enterococcus faecalis, Article, Enterococcus, Microbiology, Bacteriophage, 03 medical and health sciences, abortive infection, adsorption, rhamnopolysaccharide, enterococcus, Virology, medicine, bacterial, gene, Gene, Prophage, biology, biology.organism_classification, 3. Good health, 030104 developmental biology, Infectious Diseases, \textitEnterococcus, escherichia-coli, encodes, identification, protein, Bacteria, resistance mechanism
Abstract

Enterococcus faecalis is an opportunistic pathogen that has emerged as a major cause of nosocomial infections worldwide. Many clinical strains are indeed resistant to last resort antibiotics and there is consequently a reawakening of interest in exploiting virulent phages to combat them. However, little is still known about phage receptors and phage resistance mechanisms in enterococci. We made use of a prophageless derivative of the well-known clinical strain E. faecalis V583 to isolate a virulent phage belonging to the Picovirinae subfamily and to the P68 genus that we named Idefix. Interestingly, most isolates of E. faecalis tested&mdash, including V583&mdash, were resistant to this phage and we investigated more deeply into phage resistance mechanisms. We found that E. faecalis V583 prophage 6 was particularly efficient in resisting Idefix infection thanks to a new abortive infection (Abi) mechanism, which we designated Abi&alpha, It corresponded to the Pfam domain family with unknown function DUF4393 and conferred a typical Abi phenotype by causing a premature lysis of infected E. faecalis. The abi&alpha, gene is widespread among prophages of enterococci and other Gram-positive bacteria. Furthermore, we identified two genes involved in the synthesis of the side chains of the surface rhamnopolysaccharide that are important for Idefix adsorption. Interestingly, mutants in these genes arose at a frequency of ~10&minus, 4 resistant mutants per generation, conferring a supplemental bacterial line of defense against Idefix.

Published
2019
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12. The absence of N-acetylglucosamine in wall teichoic acids of Listeria monocytogenes modifies biofilm architecture and tolerance to rinsing and cleaning procedures

Author

Thomas Brauge, Christine Faille, Irina Sadovskaya, Alain Charbit, Thierry Benezech, Yang Shen, Martin J Loessner, Jean Romain Bautista, Graziella Midelet-Bourdin, Laboratoire de Boulogne sur mer, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Unité Matériaux et Transformations - UMR 8207 (UMET), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Supérieure de Chimie de Lille (ENSCL)-Institut de Chimie du CNRS (INC)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Institut Charles Viollette (ICV) - EA 7394 (ICV), Université d'Artois (UA)-Institut National de la Recherche Agronomique (INRA)-Université du Littoral Côte d'Opale (ULCO)-Institut Supérieur d'Agriculture-Université de Lille, Institut Necker Enfants-Malades (INEM - UM 111 (UMR 8253 / U1151)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Processus aux Interfaces et Hygiène des Matériaux (PIHM), Institut National de la Recherche Agronomique (INRA), Service des Recherches Métallurgiques Appliquées (SRMA), Département des Matériaux pour le Nucléaire (DMN), CEA-Direction des Energies (ex-Direction de l'Energie Nucléaire) (CEA-DES (ex-DEN)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-CEA-Direction des Energies (ex-Direction de l'Energie Nucléaire) (CEA-DES (ex-DEN)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Département des Produits de la Pêche et de l'Aquaculture (DPTPPA), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Supérieure de Chimie de Lille (ENSCL)-Université de Lille-Centre National de la Recherche Scientifique (CNRS), Equipe APA, Université du Littoral Côte d'Opale (ULCO), CEA-Direction de l'Energie Nucléaire (CEA-DEN), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-CEA-Direction de l'Energie Nucléaire (CEA-DEN), Laboratoire de sécurité des aliments de Maisons-Alfort, Charbit, Alain, and Institut de Chimie du CNRS (INC)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Lille-Ecole Nationale Supérieure de Chimie de Lille (ENSCL)

Subjects
0301 basic medicine, Water flow, Mutant, lcsh:Medicine, Force d'interaction, Acide téichoïque, Gene mutation, Pathology and Laboratory Medicine, Bacterial Adhesion, chemistry.chemical_compound, Cell Wall, Medicine and Health Sciences, Listeria, Biofilm, Fraction saccharidique, lcsh:Science, ComputingMilieux_MISCELLANEOUS, education.field_of_study, Teichoic acid, Multidisciplinary, Strain (chemistry), Nonsense Mutation, Bacterial Pathogens, Extracellular Matrix, Phenotype, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Medical Microbiology, Physical Sciences, Metallurgy, Pathogens, Cellular Structures and Organelles, Hydrophobic and Hydrophilic Interactions, Research Article, Materials Science, 030106 microbiology, Population, Mutagenesis (molecular biology technique), Microbiology, Acetylglucosamine, 03 medical and health sciences, Bacterial Proteins, Microscopy, Electron, Transmission, Genetics, Alloys, education, Microbial Pathogens, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology, Silent Mutation, lcsh:R, Biology and Life Sciences, Water, Bacteriology, Cell Biology, Listeria Monocytogenes, Stainless Steel, Teichoic Acids, 030104 developmental biology, chemistry, Steel, Biofilms, Mutation, lcsh:Q, Stress, Mechanical, Bacterial Biofilms
Abstract

The wall teichoic acid (WTA) is the major carbohydrate found within the extracellular matrix of the Listeria monocytogenes biofilm. We first addressed the frequency of spontaneous mutations in two genes (lmo2549 and lmo2550) responsible for the GlcNAcylation in 93 serotype 1/2a strains that were mainly isolated from seafood industries. We studied the impact of mutations in lmo2549 or lmo2550 genes on biofilm formation by using one mutant carrying a natural mutation inactivating the lmo2550 gene (DSS 1130 BFA2 strain) and two EGD-e mutants that lack respective genes by in-frame deletion of lmo2549 or lmo2550 using splicing-by-overlap-extension PCR, followed by allelic exchange mutagenesis. The lmo2550 gene mutation, occurring in around 50% isolates, caused a decrease in bacterial adhesion to stainless steel compared to wild-type EGD-e strain during the adhesion step. On the other hand, bacterial population weren’t significantly different after 24h-biofilm formation. The biofilm architecture was different between the wild-type strain and the two mutants inactivated for lmo2549 or lmo2550 genes respectively with the presence of bacterial micro-colonies for mutants which were not observed in the wild-type EGD-e strain biofilm. These differences might account for the stronger hydrophilic surface exhibited by the mutant cells. Upon a water flow or to a cleaning procedure at a shear stress of 0.16 Pa, the mutant biofilms showed the higher detachment rate compared to wild-type strain. Meanwhile, an increase in the amount of residual viable but non-culturable population on stainless steel was recorded in two mutants. Our data suggests that the GlcNAc residue of WTA played a role in adhesion and biofilm formation of Listeria monocyctogenes., PLoS ONE, 13 (1), ISSN:1932-6203

Published
2018
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13. Viability Detection of Foodborne Bacterial Pathogens in Food Environment by PMA-qPCR and by Microscopic Observation

Author

Graziella Midelet-Bourdin, Christophe Soumet, Thomas Brauge, Chiffoleau, Emmanuelle, Laboratoire de Boulogne sur mer, Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), Département des Produits de la Pêche et de l'Aquaculture (DPTPPA), Laboratoire de sécurité des aliments de Maisons-Alfort (LSAl), Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES)-Agence nationale de sécurité sanitaire de l'alimentation, de l'environnement et du travail (ANSES), and Laboratoire de Fougères, Bâtiment Bioagropolis

Subjects
bactérie pathogène alimentaire, foodborne pathogen, Microorganism, Biology, Viable but nonculturable, Microbiology, 03 medical and health sciences, Propidium monoazide, medicine, microbiologie, bacteria, sécurité des aliments, [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology, 030304 developmental biology, bactérie, 0303 health sciences, Food poisoning, 030306 microbiology, business.industry, microbiology, Biofilm, medicine.disease, Food safety, détection, Microscopic observation, food safety, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, business, Food environment
Abstract

International audience; Foodborne pathogens are responsible of foodborne diseases and food poisoning and thus pose a great threat to food safety. These microorganisms can adhere to surface and form a biofilm composed of an extracellular matrix. This extracellular matrix protects bacterial cells from industrial environmental stress factors such as cleaning and disinfection operations. Moreover, during these environmental stresses, many bacterial species can enter a viable but nonculturable (VBNC) state. VBNC cells are characterized by a loss of cultivability on conventional bacteriological agar. This leads to an underestimation of total viable cells in environmental samples, and thus poses a risk for public health. In this chapter, we present a method to detect viable population of foodborne pathogens in industrial environmental samples using a molecular method with a combination of propidium monoazide (PMA) and quantitative PCR (qPCR) and a fluorescence microscopic method associated with the LIVE/DEAD BacLight™ viability stain.

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