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1. Francisella novicida and F. philomiragia biofilm features conditionning fitness in spring water and in presence of antibiotics

Author

Bastien Touquet, Corinne Villers, Isabelle Tardieux, Patricia Renesto, Georgios Pavlou, Nandadeva Yakandawala, Claire Siebert, Thérapeutique Recombinante Expérimentale (TIMC-IMAG-TheREx ), Techniques de l'Ingénierie Médicale et de la Complexité - Informatique, Mathématiques et Applications Grenoble - UMR 5525 (TIMC-IMAG), VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA)-VetAgro Sup - Institut national d'enseignement supérieur et de recherche en alimentation, santé animale, sciences agronomiques et de l'environnement (VAS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut polytechnique de Grenoble - Grenoble Institute of Technology (Grenoble INP ), Université Grenoble Alpes (UGA), Unité de Recherche Risques Microbiens (U2RM), Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU), Membrane Dynamics of Parasite-Host Cell Interactions [Grenoble], Institute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) (IAB), Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Kane Biotech Inc. [Winnipeg, Canada], This work was supported by the FINOVI Foundation (Grant AO12-02) and by the ANR ASTRID (grant ANR-17-ASTR-0024). NY Yakandawala was employed by the company Kane Biotech, Inc. (Winnipeg, Canada)., Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre Hospitalier Universitaire [Grenoble] (CHU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang - Auvergne-Rhône-Alpes (EFS)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), and Bodescot, Myriam

Subjects
Exopolysaccharides, Microorganism, Glycobiology, Fresh Water, medicine.disease_cause, Pathology and Laboratory Medicine, Biochemistry, Tularemia, Antibiotics, Ciprofloxacin, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Medicine and Health Sciences, Francisella, Conserved Sequence, 0303 health sciences, Multidisciplinary, biology, Antimicrobials, Drugs, Adaptation, Physiological, Francisella Tularensis, Bacterial Pathogens, Anti-Bacterial Agents, Medical Microbiology, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Medicine, Pathogens, Research Article, DNA, Bacterial, Science, Microbiology, 03 medical and health sciences, Antibiotic resistance, Polysaccharides, Microbial Control, Drug Resistance, Bacterial, medicine, Humans, Francisella novicida, Microbial Pathogens, Francisella tularensis, 030304 developmental biology, Pharmacology, Bacteria, 030306 microbiology, Biofilm, Organisms, Biology and Life Sciences, Bacteriology, [SDV.EE.IEO] Life Sciences [q-bio]/Ecology, environment/Symbiosis, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, medicine.disease, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Biofilms, Antibiotic Resistance, Antimicrobial Resistance, [SDV.MP.BAC] Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, Bacterial Biofilms, Gram-Negative Bacterial Infections, [SDV.EE.IEO]Life Sciences [q-bio]/Ecology, environment/Symbiosis
Abstract

International audience; Biofilms are currently considered as a predominant lifestyle of many bacteria in nature. While they promote survival of microbes, biofilms also potentially increase the threats to animal and public health in case of pathogenic species. They not only facilitate bacteria transmission and persistence, but also promote spreading of antibiotic resistance leading to chronic infections. In the case of Francisella tularensis, the causative agent of tularemia, biofilms have remained largely enigmatic. Here, applying live and static confocal microscopy, we report growth and ultrastructural organization of the biofilms formed in vitro by these microorganisms over the early transition from coccobacillary into coccoid shape during biofilm assembly. Using selective dispersing agents, we provided evidence for extracellular DNA (eDNA) being a major and conserved structural component of mature biofilms formed by both F. subsp. novicida and a human clinical isolate of F. philomiragia. We also observed a higher physical robustness of F. novicida biofilm as compared to F. philomiragia one, a feature likely promoted by specific polysaccharides. Further, F. novicida biofilms resisted significantly better to ciprofloxacin than their planktonic counterparts. Importantly, when grown in biofilms, both Francisella species survived longer in cold water as compared to free-living bacteria, a trait possibly associated with a gain in fitness in the natural aquatic environment. Overall, this study provides information on survival of Francisella when embedded with biofilms that should improve both the future management of biofilm-related infections and the design of effective strategies to tackle down the problematic issue of bacteria persistence in aquatic ecosystems.

Published
2020
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2. Francisella novicida and F. philomiragia biofilm features conditionning fitness in spring water and in presence of antibiotics.

Author

Siebert C, Villers C, Pavlou G, Touquet B, Yakandawala N, Tardieux I, and Renesto P

Subjects
Adaptation, Physiological, Anti-Bacterial Agents pharmacology, Ciprofloxacin pharmacology, Conserved Sequence, DNA, Bacterial chemistry, Francisella drug effects, Francisella genetics, Francisella pathogenicity, Gram-Negative Bacterial Infections microbiology, Humans, Biofilms, Drug Resistance, Bacterial, Francisella physiology, Fresh Water microbiology
Abstract

Biofilms are currently considered as a predominant lifestyle of many bacteria in nature. While they promote survival of microbes, biofilms also potentially increase the threats to animal and public health in case of pathogenic species. They not only facilitate bacteria transmission and persistence, but also promote spreading of antibiotic resistance leading to chronic infections. In the case of Francisella tularensis, the causative agent of tularemia, biofilms have remained largely enigmatic. Here, applying live and static confocal microscopy, we report growth and ultrastructural organization of the biofilms formed in vitro by these microorganisms over the early transition from coccobacillary into coccoid shape during biofilm assembly. Using selective dispersing agents, we provided evidence for extracellular DNA (eDNA) being a major and conserved structural component of mature biofilms formed by both F. subsp. novicida and a human clinical isolate of F. philomiragia. We also observed a higher physical robustness of F. novicida biofilm as compared to F. philomiragia one, a feature likely promoted by specific polysaccharides. Further, F. novicida biofilms resisted significantly better to ciprofloxacin than their planktonic counterparts. Importantly, when grown in biofilms, both Francisella species survived longer in cold water as compared to free-living bacteria, a trait possibly associated with a gain in fitness in the natural aquatic environment. Overall, this study provides information on survival of Francisella when embedded with biofilms that should improve both the future management of biofilm-related infections and the design of effective strategies to tackle down the problematic issue of bacteria persistence in aquatic ecosystems., Competing Interests: The authors have declared that no competing interests exist.

Published
2020
Full Text
View/download PDF

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