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1. Response of Controlled Cell Load Biofilms to Cold Atmospheric Plasma Jet: Evidence of Extracellular Matrix Contribution

Author

Maritxu Labadie, Frédéric Marchal, Nofel Merbahi, Elisabeth Girbal-Neuhauser, Catherine Fontagné-Faucher, and Claire-Emmanuelle Marcato-Romain

Subjects
P. aeruginosa, L. citreum, bacterial survival, extracellular polymeric substances, ATR-FTIR, non-thermal plasma, Science
Abstract

Aim: Study of the biocidal effect of a cold atmospheric-pressure plasma in ambient air on single-species bacterial biofilms with controlled cell density, characterized by different extracellular matrices. Methods and results: Two bacterial strains were chosen to present different Gram properties and contrasted extracellular matrices: Pseudomonas aeruginosa ATCC 15442 (Gram-negative), and Leuconostoc citreum NRRL B-1299 (Gram-positive). P. aeruginosa biofilm exhibits a complex matrix, rich in proteins while L. citreum presents the specificity to produce glucan-type exopolysaccharides when grown in the presence of sucrose. Plasma was applied on both surface-spread cells and 24-h grown biofilms with controlled cell loads over 5, 10, or 20 min. Surface-spread bacteria showed a time dependent response, with a maximal bacterial reduction of 2.5 log after 20 min of treatment. On the other hand, in our experimental conditions, no bactericidal effect could be observed when treating biofilms of P. aeruginosa and glucan-rich L. citreum. Conclusions: For biofilms presenting equivalent cell loads, the response to plasma treatment seemed to depend on the properties of the extracellular matrix characterized by infrared spectroscopy, scanning electron microscopy, or dry weight. Significance and impact of study: Both cell load standardization and biofilm characterization are paramount factors to consider the biocide effect of plasma treatments. The extracellular matrix could affect the plasma efficacy by physical and/or chemical protective effects.

Published
2021
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2. Response of Controlled Cell Load Biofilms to Cold Atmospheric Plasma Jet: Evidence of Extracellular Matrix Contribution

Author

Frédéric Marchal, Nofel Merbahi, Catherine Fontagné-Faucher, Claire-Emmanuelle Marcato-Romain, Elisabeth Girbal-Neuhauser, Maritxu Labadie, Merbahi, Nofel, Laboratoire de Biotechnologies Agroalimentaire et Environnementale (LBAE), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Plasmas Réactifs Hors Equilibre (LAPLACE-PRHE), LAboratoire PLasma et Conversion d'Energie (LAPLACE), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3)

Subjects
P. aeruginosa, non-thermal plasma, Science, Nonthermal plasma, medicine.disease_cause, General Biochemistry, Genetics and Molecular Biology, Article, biofilm, Extracellular matrix, 03 medical and health sciences, Extracellular polymeric substance, Leuconostoc citreum, Extracellular, medicine, extracellular polymeric substances, Ecology, Evolution, Behavior and Systematics, 030304 developmental biology, 0303 health sciences, biology, 030306 microbiology, Pseudomonas aeruginosa, Chemistry, [SPI.PLASMA]Engineering Sciences [physics]/Plasmas, Biofilm, Paleontology, [SPI.PLASMA] Engineering Sciences [physics]/Plasmas, biology.organism_classification, L. citreum, [SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology, bacterial survival, ATR-FTIR, Space and Planetary Science, Biophysics, Bacteria
Abstract

Aim: Study of the biocidal effect of a cold atmospheric-pressure plasma in ambient air on single-species bacterial biofilms with controlled cell density, characterized by different extracellular matrices. Methods and results: Two bacterial strains were chosen to present different Gram properties and contrasted extracellular matrices: Pseudomonas aeruginosa ATCC 15442 (Gram-negative), and Leuconostoc citreum NRRL B-1299 (Gram-positive). P. aeruginosa biofilm exhibits a complex matrix, rich in proteins while L. citreum presents the specificity to produce glucan-type exopolysaccharides when grown in the presence of sucrose. Plasma was applied on both surface-spread cells and 24-h grown biofilms with controlled cell loads over 5, 10, or 20 min. Surface-spread bacteria showed a time dependent response, with a maximal bacterial reduction of 2.5 log after 20 min of treatment. On the other hand, in our experimental conditions, no bactericidal effect could be observed when treating biofilms of P. aeruginosa and glucan-rich L. citreum. Conclusions: For biofilms presenting equivalent cell loads, the response to plasma treatment seemed to depend on the properties of the extracellular matrix characterized by infrared spectroscopy, scanning electron microscopy, or dry weight. Significance and impact of study: Both cell load standardization and biofilm characterization are paramount factors to consider the biocide effect of plasma treatments. The extracellular matrix could affect the plasma efficacy by physical and/or chemical protective effects.

Published
2021
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3. In situ analysis of Bacillus licheniformis biofilms: amyloid-like polymers and eDNA are involved in the adherence and aggregation of the extracellular matrix

Author

Irina Randrianjatovo-Gbalou, Claire-Emmanuelle Marcato-Romain, P. Rouquette, Elisabeth Girbal-Neuhauser, and D. Lefebvre

Subjects
DNA, Bacterial, 0301 basic medicine, 030106 microbiology, Matrix (biology), Applied Microbiology and Biotechnology, Bacterial Adhesion, Extracellular matrix, 03 medical and health sciences, chemistry.chemical_compound, Biopolymers, Bacillus licheniformis, chemistry.chemical_classification, biology, Biofilm, General Medicine, Proteinase K, biology.organism_classification, Extracellular Matrix, Enzyme, Biochemistry, chemistry, Biofilms, biology.protein, Nucleic acid, DNA, Biotechnology
Abstract

Aims This study attempts to determine which of the exopolymeric substances are involved in the adherence and aggregation of a Bacillus licheniformis biofilm. Methods and Results The involvement of extracellular proteins and eDNA were particularly investigated using DNase and proteinase K treatment. The permeability of the biofilms increased fivefold after DNase I treatment. The quantification of the matrix components showed that, irrespective to the enzyme tested, eDNA and amyloid-like polymers were removed simultaneously. Size-exclusion chromatography analyses supported these observations and revealed the presence of associated nucleic acid and protein complexes in the biofilm lysates. These data suggest that some extracellular DNA and amyloid-like proteins were closely interlaced within the matrix. Finally, confocal laser scanning microscopy imaging gave supplementary clues about the 3D organization of the biofilms, confirming that eDNA and exoproteins were essentially layered under and around the bacterial cells, whereas the amyloid-like fractions were homogeneously distributed within the matrix. Conclusion These results confirm that some DNA–amyloid complexes play a key role in the modulation of the mechanical resistance of B. licheniformis biofilms. Significance and Impact of the study The study highlights the need to consider the whole structure of biofilms and to target the interactions between matrix components. A better understanding of B. licheniformis biofilm physiology and the structural organization of the matrix will strengthen strategies of biofilm control.

Published
2017
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4. A dynamic resazurin microassay allowing accurate quantification of cells and suitable for acid-forming bacteria

Author

Irina Randrianjatovo-Gbalou, Valérie Dossat-Letisse, Claire-Emmanuelle Marcato-Romain, Maritxu Labadie, Meriem Zaidi-Ait-Salem, and Catherine Fontagné-Faucher

Subjects
Microbiology (medical), Microbiology, Fluorescence, 03 medical and health sciences, chemistry.chemical_compound, Lactobacillales, Oxazines, Bacillus licheniformis, Molecular Biology, 030304 developmental biology, Colony-forming unit, Bacteriological Techniques, 0303 health sciences, Chromatography, biology, Strain (chemistry), 030306 microbiology, Resazurin, biology.organism_classification, Culture Media, Lactic acid, Xanthenes, chemistry, Acids, Bacteria
Abstract

A resazurin micro-assay was developed to quantify acidifying bacteria. The resorufin fluorescent signal was measured over time and the determined time to reach the max slope (TMS) was plotted against CFU (colony forming unit) counts. This dynamic assay enabled to quantify nine lactic acid bacteria and a Bacillus licheniformis strain despite the increasing acidity of the medium.

Published
2021
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5. Comparison of extraction methods for the characterization of extracellular polymeric substances from aggregates of three biofilm-forming phototrophic microorganisms

Author

Joséphine Leflaive, Claire-Emmanuelle Marcato-Romain, Jean-Luc Rols, Elisabeth Girbal-Neuhauser, and Emilie Loustau

Subjects
0301 basic medicine, Microorganism, 030106 microbiology, Immunology, 010501 environmental sciences, Cyanobacteria, 01 natural sciences, Applied Microbiology and Biotechnology, Microbiology, Fungal Proteins, 03 medical and health sciences, Extracellular polymeric substance, Bacterial Proteins, Rivers, Chlorophyta, Genetics, Biomass, Molecular Biology, 0105 earth and related environmental sciences, Diatoms, Phototroph, Chemistry, Extracellular Polymeric Substance Matrix, Biofilm, General Medicine, Environmental chemistry, Biofilms, Extraction methods, Water Microbiology
Abstract

This paper aims to define a robust procedure to extract extracellular polymeric substances (EPS) from aggregates of three benthic phototrophic microorganisms: the cyanobacterium Phormidium autumnale, the diatom Nitzschia palea, and the green alga Uronema confervicolum. This study focuses on the extraction efficiency of polysaccharide and protein EPS by using two physical methods (sonication, cation exchange resin) and three chemical methods (formamide, EDTA, Tween 20) with minimum cell lysis. Cell lysis was evaluated by monitoring chlorophyll a release. The results indicated that sonication or incubation of the algae aggregates with 0.25% Tween 20 induced a high level of cell lysis. A combined extraction approach, with an initial dispersing pretreatment (Ultra-Turrax, 13 500 r·min–1, 1 min), followed by formamide addition (0.22%) and then incubation with Dowex cation exchange resin (50 g per g of dry biomass), provided the highest amount of extracted EPS (mostly proteins), with low cell lysis. Furthermore, extracted EPS were characterized by size exclusion chromatography, and the obtained fingerprints revealed similar profiles for the three benthic microorganisms with a majority of low molecular weight polymers (400 to 11 300 Da). However, additional EPS of high (>600 000 Da) and intermediate (20 000 to 80 000 Da) molecular sizes were specifically detected in the diatom extracts.

Published
2018

6. Anaerobic Digestion Alters Copper and Zinc Speciation

Author

Maritxu Guiresse, Claire Emmanuelle Marcato-Romain, Emmanuel Doelsch, Samuel Legros, Clément Levard, Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Centre européen de recherche et d'enseignement des géosciences de l'environnement (CEREGE), Institut de Recherche pour le Développement (IRD)-Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Collège de France (CdF (institution))-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biotechnologies Agroalimentaire et Environnementale (LBAE), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université de Toulouse (UT)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Laboratoire Ecologie Fonctionnelle et Environnement (LEFE), Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Observatoire Midi-Pyrénées (OMP), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France -Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France -Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), DIGESTATE project, funded by ANR (Agence Nationale de la Recherche, France) [ANR-15-CE34-0003-01], U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-76SF00515], ANR-15-CE34-0003,DIGESTATE,Diagnostic des traitements des déchets et comportement des contaminants dans l'environnement(2015), Aix-Marseille Université - AMU (FRANCE), Centre de Coopération Internationale en Recherche Agronomique - CIRAD (FRANCE), Centre National de la Recherche Scientifique - CNRS (FRANCE), Collège de France (FRANCE), Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE), Institut National de la Recherche Agronomique - INRA (FRANCE), Institut de Recherche pour le Développement - IRD (FRANCE), Université Toulouse III - Paul Sabatier - UT3 (FRANCE), Centre Européen de Recherche et d'Enseignement de Géosciences de l'Environnement - CEREGE (Aix en Provence, France), Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Collège de France (CdF)-Institut national des sciences de l'Univers (INSU - CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IUT 'A' Paul Sabatier, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Laboratoire Ecologie Fonctionnelle et Environnement (ECOLAB), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Observatoire Midi-Pyrénées (OMP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), ANR: DIGESTATE,ANR-15-CE34-0003-01, Institut National Polytechnique de Toulouse - INPT (FRANCE), Aix Marseille Université (AMU)-Institut national des sciences de l'Univers (INSU - CNRS)-Collège de France (CdF (institution))-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Observatoire Midi-Pyrénées (OMP), and Météo France-Centre National d'Études Spatiales [Toulouse] (CNES)-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Météo France-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS)

Subjects
media_common.quotation_subject, Speciation, Inorganic chemistry, 0211 other engineering and technologies, chemistry.chemical_element, 02 engineering and technology, Zinc, 010501 environmental sciences, 01 natural sciences, Water Purification, chemistry.chemical_compound, Bioreactors, Anaerobic digestion, Environmental Chemistry, 0105 earth and related environmental sciences, media_common, 2. Zero hunger, 021110 strategic, defence & security studies, Sewage, [SDE.IE]Environmental Sciences/Environmental Engineering, 000 - Autres thèmes, Géochimie, Zinc phosphate, Q70 - Traitement des déchets agricoles, General Chemistry, Biodegradable waste, Contamination, Copper, Amorphous solid, Chimie organique, X-Ray Absorption Spectroscopy, chemistry, 13. Climate action, Metals, Environmental chemistry, [CHIM.OTHE]Chemical Sciences/Other
Abstract

International audience; Anaerobic digestion is a widely used organic waste treatment process. However, little is known on how it could alter the speciation of contaminants in organic waste. This study was focused on determining the influence of anaerobic digestion on the speciation of copper and zinc, two metals that generally occur at high concentration in organic waste. Copper and zinc speciation was investigated by X-ray absorption spectroscopy in four different raw organic wastes (predigestion) and their digested counterparts (postdigestion, i.e., digestates). The results highlighted an increase in the digestates of the proportion of amorphous or nanostructured copper sulfides as well as amorphous or nanostructured zinc sulfides and zinc phosphate as compared to raw waste. We therefore suggest that the environmental fate of these elements would be different when spreading either digestates or raw waste on cropland.

Published
2017
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7. Combining hydrodynamic and enzymatic treatments to improve multi-species thick biofilm removal

Author

Yolaine Bessiere, Claire-Emmanuelle Marcato-Romain, Elisabeth Girbal-Neuhauser, Isabelle Queinnec, Yoan Pechaud, Etienne Paul, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire de Biotechnologies Agroalimentaire et Environnementale (LBAE), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Équipe Méthodes et Algorithmes en Commande (LAAS-MAC), Laboratoire d'analyse et d'architecture des systèmes (LAAS), Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Agence Nationale pour la Recherche [ANR-08-ECOT-003], Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IUT 'A' Paul Sabatier, Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse 1 Capitole (UT1), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse 1 Capitole (UT1), Université Toulouse III - Paul Sabatier (UPS), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UPS), Centre National de la Recherche Scientifique (CNRS), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université de Toulouse (UT)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Université Toulouse Capitole (UT Capitole), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université de Toulouse (UT)-Université Toulouse Capitole (UT Capitole)

Subjects
Detachment, [SDV]Life Sciences [q-bio], General Chemical Engineering, Transport processes, Heterotroph, Industrial and Manufacturing Engineering, Diffusion, Streptococcus mutans, 03 medical and health sciences, Paper industry, Mass transfer, Shear stress, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, 030304 developmental biology, 0303 health sciences, SHEAR-STRESS, Fouling, Biot number, 030306 microbiology, Chemistry, Applied Mathematics, Environmental engineering, Biofilm, General Chemistry, Penetration (firestop), Pulp and paper industry, STREPTOCOCCUS-MUTANS, 6. Clean water, Bacterial biofilms, Enzyme, Hydrodynamics, Cohesion, Biofilm removal, Strength, Aeration, PAPER-INDUSTRY
Abstract

The high water recycling ratio used in some industries can cause nutrients to accumulate in the system, leading to thick, rapidly growing biofilms inducing detrimental effects. Nowadays, chemical treatments are widely used but the increasingly restrictive environmental regulations induce the necessity to take the environmental aspect into account in the choice of a suitable treatment. With this in mind, this study analyses a method for controlling thick, dense biofilm development by applying synergistic actions having low environmental impact, i.e., an enzymatic and a mechanical (shear stress) treatment well suited to industrial water networks. For that purpose, biofilms were grown on plastic plates set in a Couette-Taylor reactor (CTR) that was inoculated using a "white water" sampled from a paper industry network. Development of the biofilms was controlled by applying a constant continuous feeding with a high COD/N ratio of 20, a well-defined shear stress and an external aeration, those conditions leading to a well-controlled G-value and a gamma(S/O2) value close to 1, and consequently, to the development of thick mainly aerobic heterotrophic biofilms representative of that encountered in high loaded industrial water networks. Optimal operating conditions for biofilm treatments were determined considering the penetration time for the enzymatic treatment taking into account both internal and external mass transport (using Biot number calculation). In addition, an optimal shear stress increment of 2.5 Pa was selected in order to maximise biofilm detachment while avoiding compression phenomena due to the mechanical treatment. The combined treatment led to an increase of 80% in biofilm mass removal (COD) compared to the enzymatic treatment alone and removed a large part of the basal layer of the biofilm, 80% reduction being observed in the support coverage. (C) 2012 Elsevier Ltd. All rights reserved.

Published
2012
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8. Quantification of biofilm exopolysaccharides using an in situ assay with periodic acid-Schiff reagent

Author

Elisabeth Girbal-Neuhauser, Claire-Emmanuelle Marcato-Romain, and Irina Randrianjatovo-Gbalou

Subjects
0301 basic medicine, 030106 microbiology, Biophysics, Biology, Polysaccharide, medicine.disease_cause, Biochemistry, 03 medical and health sciences, chemistry.chemical_compound, Polysaccharides, Extracellular, medicine, Rosaniline Dyes, Bacillus licheniformis, Molecular Biology, chemistry.chemical_classification, Pseudomonas aeruginosa, Periodic Acid, Biofilm, Periodic acid, Cell Biology, biology.organism_classification, 030104 developmental biology, Dextran, chemistry, Reagent, Biofilms
Abstract

A novel approach to the quantification of extracellular polysaccharides in miniaturized biofilms presenting a wide variety of extracellular matrices was developed. The assay used the periodic acid-Schiff reagent and was first calibrated on dextran and alginate solutions. Then it was implemented on 24-h and 48-h biofilms from three strains known to produce different exopolymeric substances (Pseudomonas aeruginosa, Bacillus licheniformis, Weissella confusa). The assay allowed quantification of the total exopolysaccharides, taking into account possible interferences due to cells or other main expolymers of the matrix (eDNA, proteins).

Published
2015

9. Quantification of amyloid fibrils using size exclusion chromatography coupled with online fluorescence and ultraviolet detection

Author

Elisabeth Girbal-Neuhauser, Claire-Emmanuelle Marcato-Romain, and Irina Randrianjatovo-Gbalou

Subjects
Models, Molecular, Amyloid, Calibration curve, Size-exclusion chromatography, Biophysics, Bacillus, Fractionation, Biochemistry, Protein Aggregation, Pathological, Absorbance, chemistry.chemical_compound, Protein Aggregates, Bacterial Proteins, mental disorders, Animals, Benzothiazoles, Molecular Biology, Fluorescent Dyes, Chromatography, Biofilm, Caseins, Cell Biology, Fluorescence, Molecular Weight, Thiazoles, Spectrometry, Fluorescence, chemistry, Solubility, Biofilms, Calibration, Chromatography, Gel, Thioflavin, Cattle, Spectrophotometry, Ultraviolet, France, Algorithms
Abstract

An amyloid fibrils investigation within biofilm samples requires distinguishing the amyloid β-sheet structure of these proteins and quantifying them. In this study, the property of amyloids to incorporate the fluorescent dye Thioflavin T has been exploited to propose a method of quantification. The experimental protocol includes the preparation of amyloids from commercial κ-casein (κCN) and their fractionation through size exclusion chromatography (SEC) to provide calibration curves from fluorescence and absorbance signals. Finally, a bacterial biofilm extract was injected into SEC, and the amyloid fibrils could be expressed as equivalent κCN, representing approximately 21% of the total proteins.

Published
2015

10. Development and validation of a colorimetric assay for simultaneous quantification of neutral and uronic sugars

Author

Caroline Rondel, Claire-Emmanuelle Marcato-Romain, and Elisabeth Girbal-Neuhauser

Subjects
Environmental Engineering, Serum albumin, Uronic acid, Polysaccharide, Anthrone, Sensitivity and Specificity, Absorbance, Matrix (chemical analysis), chemistry.chemical_compound, Bacterial Proteins, Glucuronic Acid, Bovine serum albumin, Waste Management and Disposal, Water Science and Technology, Civil and Structural Engineering, chemistry.chemical_classification, Chromatography, biology, Ecological Modeling, Polysaccharides, Bacterial, Serum Albumin, Bovine, Glucuronic acid, Pollution, Glucose, chemistry, biology.protein, Colorimetry
Abstract

A colorimetric assay based on the conventional anthrone reaction was investigated for specific quantification of uronic acids (UA) in the presence of neutral sugars and/or proteins. Scanning of glucose (Glu) and glucuronic acid (GlA) was performed after the reaction with anthrone and a double absorbance reading was made, at 560 nm and at 620 nm, in order to quantify the UA and neutral sugars separately. The assay was implemented on binary or ternary solutions containing Glu, GlA and bovine serum albumin (BSA) in order to validate its specificity towards sugars and check possible interference with other biochemical components such as proteins. Statistical analysis indicated that this assay provided correct quantification of uronic sugars from 50 to 400 mg/l and of neutral sugars from 20 to 80 mg/l, in the presence of proteins with concentrations reaching 600 mg/l. The proposed protocol can be of great interest for simultaneous determination of uronic and neutral sugars in complex biological samples. In particular, it can be used to correctly quantify the Extracellular Polymeric Substances (EPS) isolated from the biological matrix of many bacterial aggregates, even in the presence of EPS extractant such as EDTA.

Published
2012

11. Removal of microbial multi-species biofilms from the paper industry by enzymatic treatments

Author

Claire-Emmanuelle Marcato-Romain, Elisabeth Girbal-Neuhauser, Valérie Dossat-Letisse, Etienne Paul, Yoan Pechaud, Laboratoire de Biotechnologies Agroalimentaire et Environnementale (LBAE), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), 'Agence Nationale pour la Recherche' under the PRECODD [ANR-08-ECOT-003], Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IUT 'A' Paul Sabatier, Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire de Biotechnologies Agroalimentaire et Environnementale ( LBAE ), Université Toulouse III - Paul Sabatier ( UPS ), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier ( UPS ), Lab Biotechnol Agroalimentaire & Environm, Institut Universitaire de Technologie, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés ( LISBP ), Institut National de la Recherche Agronomique ( INRA ) -Institut National des Sciences Appliquées - Toulouse ( INSA Toulouse ), Institut National des Sciences Appliquées ( INSA ) -Institut National des Sciences Appliquées ( INSA ) -Centre National de la Recherche Scientifique ( CNRS ), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université de Toulouse (UT)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), and ANR-08-ECOT-0003,BIOFIME,Maîtrise Intégrée des Biofilms papetiers par actions combinées à Faible Impact Environnemental(2008)

Subjects
Paper, Proteases, medicine.medical_treatment, enzymes, PROTEIN, Multi-methods extraction, Aquatic Science, Polysaccharide, EXTRACELLULAR P, Applied Microbiology and Biotechnology, extracellular polymeric substances (EPS), EXTRACELLULAR POLYMERIC SUBSTANCES, 03 medical and health sciences, Hydrolysis, Industrial Microbiology, Extracellular polymeric substance, BIOFLOCCULATION, MILL, medicine, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, SLIME, 030304 developmental biology, Water Science and Technology, Extracellular Polymeric Substances EPS, chemistry.chemical_classification, 0303 health sciences, Protease, [ SDV ] Life Sciences [q-bio], biology, Bacteria, 030306 microbiology, [SDE.IE]Environmental Sciences/Environmental Engineering, enzymatic removal, Biofilm, biology.organism_classification, Pulp and paper industry, paper mill biofilms, Extracellular Matrix, Enzyme, chemistry, Biofilms, Sugar composition, Peptide Hydrolases
Abstract

International audience; This study aimed to characterize biofilms from the paper industry and evaluate the effectiveness of enzymatic treatments in reducing them. The extracellular polymeric substances (EPS) extracted from six industrial biofilms were studied. EPS were mainly proteins, the protein to polysaccharide ratio ranging from 1.3 to 8.6 depending on where the sampling point was situated in the paper making process. Eight hydrolytic enzymes were screened on a 24-h multi-species biofilm. The enzymes were tested at various concentrations and contact durations. Glycosidases and lipases were inefficient or only slightly efficient for biofilm reduction, while proteases were more efficient: after treatment for 24 h with pepsin, Alcalase® or Savinase®, the removal exceeded 80%. Savinase® appeared to be the most adequate for industrial conditions and was tested on an industrial biofilm sample. This enzyme led to a significant release of proteins from the EPS matrix, indicating its potential efficiency on an industrial scale.

Published
2012
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12. Coupling hydrodynamics and bio-chemical treatment: an alternative to improve thick biofilm removal

Author

Yoan Pechaud, Yolaine Bessiere, Claire-Emmanuelle Marcato-Romain, Valérie Dossat-Letisse, Isabelle Queinnec, Etienne Paul, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire de Biotechnologies Agroalimentaire et Environnementale (LBAE), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IUT 'A' Paul Sabatier, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Équipe Méthodes et Algorithmes en Commande (LAAS-MAC), Laboratoire d'analyse et d'architecture des systèmes (LAAS), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université de Toulouse (UT)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Université Toulouse Capitole (UT Capitole), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université Toulouse Capitole (UT Capitole), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse 1 Capitole (UT1), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse 1 Capitole (UT1)

Subjects
[SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering
Published
2011

13. Improving fighting against biofilms by combining hydrodynamic and enzymatic treatments

Author

Yoan Pechaud, Yolaine Bessiere, Claire-Emmanuelle Marcato-Romain, Valérie Dossat-Letisse, Isabelle Queinnec, Etienne Paul, Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Laboratoire de Biotechnologies Agroalimentaire et Environnementale (LBAE), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IUT 'A' Paul Sabatier, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées, Équipe Méthodes et Algorithmes en Commande (LAAS-MAC), Laboratoire d'analyse et d'architecture des systèmes (LAAS), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse 1 Capitole (UT1)-Université Toulouse - Jean Jaurès (UT2J), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse 1 Capitole (UT1), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Université Toulouse 1 Capitole (UT1), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse 1 Capitole (UT1), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut Universitaire de Technologie - Paul Sabatier (IUT Paul Sabatier), Université de Toulouse (UT)-Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT), Université Toulouse Capitole (UT Capitole), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université Toulouse - Jean Jaurès (UT2J), Université de Toulouse (UT)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université de Toulouse (UT)-Université Toulouse Capitole (UT Capitole)

Subjects
[SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering
Published
2011

14. New direct contact approach to evaluate soil genotoxicity using the Vicia faba micronucleus test

Author

Maritxu Guiresse, Claire-Emmanuelle Marcato-Romain, Eric Pinelli, Sylvie Cotelle, Marie Cecchi, Laboratoire Ecologie Fonctionnelle et Environnement (ECOLAB), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Observatoire Midi-Pyrénées (OMP), Météo France-Centre National d'Études Spatiales [Toulouse] (CNES)-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Météo France-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Centre National de la Recherche Scientifique (CNRS), Laboratoire des Interactions Ecotoxicologie, Biodiversité, Ecosystèmes (LIEBE), Centre National de la Recherche Scientifique (CNRS)-Université Paul Verlaine - Metz (UPVM), Centre National de la Recherche Scientifique - CNRS (FRANCE), Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE), Université Toulouse III - Paul Sabatier - UT3 (FRANCE), Université Paul-Verlaine - Metz (FRANCE), Laboratoire Ecologie Fonctionnelle et Environnement (LEFE), Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Observatoire Midi-Pyrénées (OMP), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France -Institut de Recherche pour le Développement (IRD)-Institut national des sciences de l'Univers (INSU - CNRS)-Centre National d'Études Spatiales [Toulouse] (CNES)-Centre National de la Recherche Scientifique (CNRS)-Météo-France -Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT), and Université Paul Verlaine - Metz (UPVM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Environmental Engineering, Health, Toxicology and Mutagenesis, Biodiversité et Ecologie, 0211 other engineering and technologies, 02 engineering and technology, 010501 environmental sciences, medicine.disease_cause, Ecotoxicology, 01 natural sciences, complex mixtures, Soil, [SDU.STU.GC]Sciences of the Universe [physics]/Earth Sciences/Geochemistry, medicine, Environmental Chemistry, Soil Pollutants, Polycyclic Aromatic Hydrocarbons, ComputingMilieux_MISCELLANEOUS, 0105 earth and related environmental sciences, Risk assessment, 2. Zero hunger, 021110 strategic, defence & security studies, Micronucleus Tests, Chemistry, Public Health, Environmental and Occupational Health, Géochimie, Sowing, General Medicine, General Chemistry, PAH, 15. Life on land, Pollution, Soil quality, Solid medium, Vicia faba, Lead, Environmental chemistry, Soil water, Micronucleus test, Soil fertility, [SDE.BE]Environmental Sciences/Biodiversity and Ecology, Micronucleus, Genotoxicity, Environmental Monitoring, Mutagens
Abstract

International audience; A method to assess micronucleus (MN) induction in Vicia faba roots by direct contact exposure to a solid matrix was developed. The procedure comprised a 5-d germination period, as in the well-known method using aqueous extracts. However, the seeds were here sown directly into the test soil whereas a culture period is necessary before exposing seedlings to a liquid medium. One soil under forest and two contaminated soils from areas affected by industrial installations and a coke works were used. Three durations of direct exposure were tested: 2, 5 and 7 d. The optimal duration was evaluated at 2 d to observe maximal MN induction without observing toxicity symptoms. The methodology using aqueous extracts was applied to the same three soils: MN frequency was higher than in the direct contact assay but the ratios of MN frequencies from tested soils in comparison to the negative control were lower. However, for each soil, both the direct contact method and the aqueous extract exposure led to the same risk assessment diagnosis. The evaluation of a concentration range of a polycyclic aromatic hydrocarbons (PAH)-contaminated soil showed a dose-dependent MN frequency when the seeds were allowed to germinate before sowing in the soil: the soil genotoxicity was the highest at intermediate doses. The direct contact method was found to be rapid, sensitive and well suited to the evaluation of soil quality

Published
2009
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