Utilisation post-récolte de sels organiques et inorganiques pour lutter contre la pourriture molle de la pomme de terre : base physico-chimique

Le but de cette thèse était d’évaluer le potentiel antimicrobien de 21 sels organiques et inorganiques pour contrôler la pourriture molle de la pomme de terre, causée par les bactéries Erwinia carotovora subsp. carotovora et Erwinia carotovora subsp. atroseptica. Les résultats obtenus ont montré que 11 sels ont complètement inhibé la croissance bactérienne in vitro, parmi lesquels le carbonate de sodium, le métabisulfite de sodium, le phosphate de sodium tribasique, le lactate d’aluminium, le chlorure d’aluminium, le bicarbonate de sodium, l’acétate d’ammonium, le di-hydroxy acétate d’aluminium, le sorbate de potassium et le benzoate de sodium se sont avérés bactéricides. Appliqués sur les tubercules de pomme de terre, seuls le chlorure d’aluminium, le métabisulfite de sodium et dans une moindre mesure le benzoate de sodium ont permis de réduire la sévérité de la maladie, et ce de façon curative et préventive. Il semble à la lumière des résultats obtenus, que l’effet inhibiteur des sels observé in vitro résulte de la capacité d’ionisation de l’eau (faible pKa ou pKb) de leurs constituants, ainsi que de leur lipophilicité (dans le cas du benzoate de sodium et du sorbate de potassium). Cette capacité d’ioniser l’eau des ions serait cependant altérée par le pouvoir tampon du tubercule. Pour leur part, les sels efficaces contre la maladie disposeraient de caractéristiques spécifiques leur permettant de demeurer actifs dans le tissu. Des études ultrastructurales ont montré que le chlorure d’aluminium provoque le bris de l’enveloppe bactérienne, une disparition des vésicules extracellulaires et une agrégation du cytoplasme, phénomènes non observés chez les bactéries traitées avec le métabisulfite de sodium. Enfin, l’évaluation de l’effet de certains sels sur la qualité des tubercules a montré d’une part que la perte de poids a été généralement plus importante chez les tubercules traités au chlorure d’aluminium ou au métabisulfite de sodium, et d’autre part que l’application des sels organiques, particulièrement de benzoate de sodium, a favorisé une augmentation du contenu en sucres solubles chez les tubercules. Cette étude démontre que le chlorure d’aluminium et le métabisulfite de sodium pourraient être mis à profit pour la conservation de la pomme de terre.The objective of this thesis was to evaluate the antimicrobial potential of 21 organic and inorganic salts to control Erwinia carotovora subsp. carotovora (Ecc) and Erwinia carotovora subsp. atroseptica (Eca), two bacteria responsible for soft rot development in stored potato tubers. The results have shown that eleven salts (at 0.2 M) have completely inhibited the bacterial growth in vitro, among which sodium carbonate, sodium metabisulfite, trisodium phosphate, aluminum lactate, aluminum chloride, sodium bicarbonate, ammonium acetate, aluminum di-hydroxy-acetate, potassium sorbate and sodium benzoate were bactericidal. On potato tubers, only aluminum chloride, sodium metabisulfite and to a lesser extent sodium benzoate have markedly controlled the disease severity, during both curative and preventive applications. Based on the obtained results, it appears that the inhibitory effect of salts in vitro relates to the water ionization capacity (low pKa or low pKb) of their constituent ions, as well as their lipophilicity. However, the effect was dampened in vivo, presumably due to the buffering capacity of tuber tissue and Donnan effect which could lead to generation of less effective ionic species. It is likely that those salts effective in vivo exhibit other properties contributing to their effectiveness. Ultrastructural studies on Eca showed that aluminum chloride caused rupture of bacterial envelope, and cytoplasmic aggregation, which were not observed in the bacteria treated with sodium metabisulfite. These observations suggest that a part of the toxic effect of aluminum chloride originates from alterations of the bacterial envelope, whereas rapid bacterial death caused by sodium metabisulfite occurs intracellularly through interaction with biomolecules, facilitated by the diffusion of SO2. Finally, evaluation of the effect of salts on tuber quality revealed that tuber weight loss was generally higher with aluminum chloride and sodium metabisulfite treatments, while the organic salts, particularly sodium benzoate, increased sugar content of the tubers. It was concluded that aluminum chloride and sodium metabisulfite can be profitably used to fight potato storage soft rot

Anti-bacterial activity of inorganic nanomaterials and their antimicrobial peptide conjugates against resistant and non-resistant pathogens

This review details the antimicrobial applications of inorganic nanomaterials of mostly metallic form, and the augmentation of activity by surface conjugation of peptide ligands. The review is subdivided into three main sections, of which the first describes the antimicrobial activity of inorganic nanomaterials against gram-positive, gram-negative and multidrug-resistant bacterial strains. The second section highlights the range of antimicrobial peptides and the drug resistance strategies employed by bacterial species to counter lethality. The final part discusses the role of antimicrobial peptide-decorated inorganic nanomaterials in the fight against bacterial strains that show resistance. General strategies for the preparation of antimicrobial peptides and their conjugation to nanomaterials are discussed, emphasizing the use of elemental and metallic oxide nanomaterials. Importantly, the permeation of antimicrobial peptides through the bacterial membrane is shown to aid the delivery of nanomaterials into bacterial cells. By judicious use of targeting ligands, the nanomaterial becomes able to differentiate between bacterial and mammalian cells and, thus, reduce side effects. Moreover, peptide conjugation to the surface of a nanomaterial will alter surface chemistry in ways that lead to reduction in toxicity and improvements in biocompatibility

Physicochemical Basis for the Inhibitory Effects of Organic and Inorganic Salts on the Growth of Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum▿

Twenty-one salts were tested for their effects on the growth of Pectobacterium carotovorum subsp. carotovorum and Pectobacterium atrosepticum. In liquid medium, 11 salts (0.2 M) exhibited strong inhibition of bacterial growth. The inhibitory action of salts relates to the water-ionizing capacity and the lipophilicity of their constituent ions

Ultrastructural Alterations of Erwinia carotovora subsp. atroseptica Caused by Treatment with Aluminum Chloride and Sodium Metabisulfite

Aluminum and bisulfite salts inhibit the growth of several fungi and bacteria, and their application effectively controls potato soft rot caused by Erwinia carotovora. In an effort to understand their inhibitory action, ultrastructural changes in Erwinia carotovora subsp. atroseptica after exposure (0 to 20 min) to different concentrations (0.05, 0.1, and 0.2 M) of these salts were examined by using transmission electron microscopy. Plasma membrane integrity was evaluated by using the SYTOX Green fluorochrome that penetrates only cells with altered membranes. Bacteria exposed to all aluminum chloride concentrations, especially 0.2 M, exhibited loosening of the cell walls, cell wall rupture, cytoplasmic aggregation, and an absence of extracellular vesicles. Sodium metabisulfite caused mainly a retraction of plasma membrane and cellular voids which were more pronounced with increasing concentration. Bacterial mortality was closely associated with SYTOX stain absorption when bacteria were exposed to either a high concentration (0.2 M) of aluminum chloride or prolonged exposure (20 min) to 0.05 M aluminum chloride or to a pH of 2.5. Bacteria exposed to lower concentrations of aluminum chloride (0.05 and 0.1 M) for 10 min or less, or to metabisulfite at all concentrations, did not exhibit significant stain absorption, suggesting that no membrane damage occurred or it was too weak to allow the penetration of the stain into the cell. While mortality caused by aluminum chloride involves membrane damage and subsequent cytoplasmic aggregation, sulfite exerts its effect intracellularly; it is transported across the membrane by free diffusion of molecular SO(2) with little damage to the cellular membrane

Sensor Kinase RscS Induces the Production of Antigenically Distinct Outer Membrane Vesicles That Depend on the Symbiosis Polysaccharide Locus in Vibrio fischeri

Robust biofilm formation by Vibrio fischeri depends upon activation of the symbiosis polysaccharide (syp) locus, which is achieved by overexpressing the RscS sensor kinase (RscS+). Other than the Syp polysaccharide, however, little is known about V. fischeri biofilm matrix components. In other bacteria, biofilms contain polysaccharides, secreted proteins, and outer membrane vesicles (OMVs). Here, we asked whether OMVs are part of V. fischeri biofilms. Transmission electron microscopy revealed OMV-like particles between cells within colonies. In addition, OMVs could be purified from culture supernatants of both RscS+ and control cells, with the former releasing 2- to 3-fold more OMVs. The increase depended upon the presence of an intact syp locus, as an RscS+ strain deleted for sypK, which encodes a putative oligosaccharide translocase, exhibited reduced production of OMVs; it also showed a severe defect in biofilm formation. Western immunoblot analyses revealed that the RscS+ strain, but not the control strain or the RscS+ sypK mutant, produced a distinct set of nonproteinaceous molecules that could be detected in whole-cell extracts, OMV preparations, and lipopolysaccharide (LPS) extracts. Finally, deletion of degP, which in other bacteria influences OMV production, decreased OMV production and reduced the ability of the cells to form biofilms. We conclude that overexpression of RscS induces OMV production in a manner that depends on the presence of the syp locus and that OMVs produced under these conditions contain antigenically distinct molecules, possibly representing a modified form of lipopolysaccharide (LPS). Finally, our data indicate a correlation between OMV production and biofilm formation by V. fischeri