1. Next-Generation Sequencing and MALDI Mass Spectrometry in the Study of Multiresistant Processed Meat Vancomycin-Resistant Enterococci (VRE)
- Author
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Gilberto Igrejas, Richard Bonnet, Christophe Chambon, Michel Hébraud, Patrícia Poeta, Racha Beyrouthy, Laetitia Théron, Didier Viala, Telma de Sousa, Soraia Oliveira, Manuela Caniça, Carolina Sabença, University of Trás-os-Montes and Alto Douro [Portugal] (UTAD), Laboratorio Associado Para a Quimica Verde (LAQV/REQUIMTE), Universidade Nova de Lisboa = NOVA University Lisbon (NOVA), Unité Mixte de Recherche sur les Herbivores - UMR 1213 (UMRH), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Qualité des Produits Animaux (QuaPA), Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Plateforme Exploration du Métabolisme (PFEM), Institut National de la Recherche Agronomique (INRA)-Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-MetaboHUB-Clermont, MetaboHUB-MetaboHUB, Microbiologie Environnement Digestif Santé (MEDIS), Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Centre National de Référence de la Résistance aux Antibiotiques (CNR), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Microbes, Intestin, Inflammation et Susceptibilité de l'Hôte (M2iSH), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Centre de Recherche en Nutrition Humaine d'Auvergne (CRNH d'Auvergne)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), National Reference Laboratory of Antibiotic Resistances and Healthcare Associated Infections (NRL-AMR-HAI), Department of Infectious Diseases, Instituto Nacional de Saùde Dr Ricardo Jorge [Portugal] (INSA), Faculty of Science Oporto University, Centre for the Studies of Animal Science, Institute of Agrarian and Agri-Food Sciences and Technologies, Oporto University, 4051-401 Oporto, 'Espectrometria de massa e sequenciacao aplicadas a microbiologia clinica moderna: pesquisa e identificacao de biomarcadores em especies bacterianas multiresistentes' (Programa de Accoes Universitarias Integradas Luso-Francesas) TC-14/2017Associate Laboratory for Green Chemistry, LAQV - FCT/MCTES UID/QUI/50006/2020CT/MCTES UIDB/00211/2020, LAQV@REQUIMTE, Plateforme Exploration du Métabolisme-MetaboHUB (PFEM-MetaboHUB), Institut National de la Recherche Agronomique (INRA)-Université Blaise Pascal - Clermont-Ferrand 2 (UBP), ROSSI, Sabine, VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and INRAE, Plateforme d’Exploration du Métabolisme, Composante Protéomique (PFEMcp), 63122 Saint-Genès Champanelle
- Subjects
0301 basic medicine ,antibiotic resistance ,Tetracycline ,Antibiotic resistance ,[SDV]Life Sciences [q-bio] ,030106 microbiology ,Processed meat ,General Biochemistry, Genetics and Molecular Biology ,Article ,Microbiology ,03 medical and health sciences ,Immunology and Microbiology(all) ,medicine ,MALDI-TOF MS ,lcsh:QH301-705.5 ,Enterococcus spp ,processed meat ,General Immunology and Microbiology ,biology ,Agricultural and Biological Sciences(all) ,Resistência aos Antimicrobianos ,Teicoplanin ,Biochemistry, Genetics and Molecular Biology(all) ,Kanamycin ,biochemical phenomena, metabolism, and nutrition ,biology.organism_classification ,3. Good health ,[SDV] Life Sciences [q-bio] ,030104 developmental biology ,Enterococcus ,lcsh:Biology (General) ,Streptomycin ,Next-generation sequencing ,Gentamicin ,next-generation sequencing ,General Agricultural and Biological Sciences ,medicine.drug ,Enterococcus faecium - Abstract
Vancomycin-resistant enterococci (VRE), due to their intrinsic resistance to various commonly used antibiotics and their malleable genome, make the treatment of infections caused by these bacteria less effective. The aims of this work were to characterize isolates of Enterococcus spp. that originated from processed meat, through phenotypic and genotypic techniques, as well as to detect putative antibiotic resistance biomarkers. The 19 VRE identified had high resistance to teicoplanin (89%), tetracycline (94%), and erythromycin (84%) and a low resistance to kanamycin (11%), gentamicin (11%), and streptomycin (5%). Based on a Next-Generation Sequencing NGS technique, most isolates were vanA-positive. The most prevalent resistance genes detected were erm(B) and aac(6')-Ii, conferring resistance to the classes of macrolides and aminoglycosides, respectively. MALDI-TOF mass spectrometry (MS) analysis detected an exclusive peak of the Enterococcus genus at m/z (mass-to-charge-ratio) 4428 ±, 3, and a peak at m/z 6048 ±, 1 allowed us to distinguish Enterococcus faecium from the other species. Several statistically significant protein masses associated with resistance were detected, such as peaks at m/z 6358.27 and m/z 13237.3 in ciprofloxacin resistance isolates. These results reinforce the relevance of the combined and complementary NGS and MALDI-TOF MS techniques for bacterial characterization.
- Published
- 2020
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