STRESS HORMONES INCREASE BIOFILM FORMATION OF ENTEROCOCCUS FAECALIS

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Open-resorcinarenes, a new family of multivalent scaffolds

A new family of multivalent ligand platforms, the open-resorcinarenes, has been prepared in a straightforward two-step reaction. Modification of the core gives a range of topologically diverse scaffolds; functionalisation confirms the versatility of this approach, as shown through the formation of an octacalixarene array

Activation of the cell wall stress response in pseudomonas aeruginosa infected by a pf4 phage variant

Pseudomonas aeruginosa PAO1 has an integrated Pf4 prophage in its genome, encoding a relatively well-characterized filamentous phage, which contributes to the bacterial biofilm organization and maturation. Pf4 variants are considered as superinfectives when they can re-infect and kill the prophage-carrying host. Herein, the response of P. aeruginosa H103 to Pf4 variant infection was investigated. This phage variant caused partial lysis of the bacterial population and modulated H103 physiology. We show by confocal laser scanning microscopy that a Pf4 variant-infection altered P. aeruginosa H103 biofilm architecture either in static or dynamic conditions. Interestingly, in the latter condition, numerous cells displayed a filamentous morphology, suggesting a link between this phenotype and flow-related forces. In addition, Pf4 variant-infection resulted in cell envelope stress response, mostly mediated by the AlgU and SigX extracytoplasmic function sigma factors (ECFσ). AlgU and SigX involvement may account, at least partly, for the enhanced expression level of genes involved in the biosynthesis pathways of two matrix exopolysaccharides (Pel and alginates) and bis-(3′-5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) metabolism

Fixing the conformation of calix[4]arenes: When are three carbons not enough?

Calix[4]arenes are unique macrocycles that through judicious functionalisation at the lower-rim can be either fixed in one of four conformations or remain conformationally flexible. Introduction of propynyl or propenyl groups unexpectedly provides a new possibility; a unidirectional conformational switch, with the 1,3-alternate and 1,2-alternate conformers switching to the partial cone conformation, whilst the cone conformation is unchanged, under standard experimental conditions. Using 1H NMR kinetic studies, rates of switching have been shown to be dependent on the starting conformation, upper-rim substituent, where reduction in bulk enables faster switching, solvent and temperature with 1,2-alternate conformations switching fastest. Ab initio calculations (DFT) confirmed the relative stabilities of the conformations and point towards the partial cone conformer being the most stable of the four. The potential impact on synthesis through the ‘click’ reaction has been investigated and found not to be significant

Probiotic Potential and Safety Evaluation of Enterococcus faecalis OB14 and OB15, Isolated From Traditional Tunisian Testouri Cheese and Rigouta, Using Physiological and Genomic Analysis

Lactic acid bacteria (LAB) strains OB14 and OB15 were isolated from traditional Tunisian fermented dairy products, Testouri cheese and Rigouta, respectively. They were identified as Enterococcus faecalis by the MALDI TOF-MS (matrix assisted laser desorption-ionization time of flight mass spectrometry) biotyper system and molecular assays (species-specific PCR). These new isolates were evaluated for probiotic properties, compared to E. faecalis Symbioflor 1 clone DSM 16431, as reference. The bacteria were found to be tolerant to the harsh conditions of the gastrointestinal tract (acidity and bile salt). They were low to moderate biofilm producers, can adhere to Caco-2/TC7 intestinal cells and strengthen the intestinal barrier through the increase of the transepithelial electrical resistance (TER). Susceptibility to ampicillin, vancomycin, gentamicin and erythromycin has been tested using the broth microdilutions method. The results demonstrated that E. faecalis OB14 and OB15 were sensitive to the clinically important ampicillin (MIC = 1 μg/mL) and vancomycin (MIC = 2 μg/mL) antibiotics. However, Whole Genome Sequencing (WGS) showed the presence of tetracycline resistance and cytolysin genes in E. faecalis OB14, and this led to high mortality of Galleria Mellonella larvae in the virulence test. Hierarchical cluster analysis by MALDI TOF-MS biotyper showed that E. faecalis OB15 was closely related to the E. faecalis Symbioflor 1 probiotic strain than to OB14, and this has been confirmed by WGS using the average nucleotide identity (ANI) and Genome-to-Genome Hybridization similarity methods. According to these results, E. faecalis OB15 seems to be reliable for future development as probiotic, in food or feed industry

Sources, diversity and adhesion properties of Pseudomonas aeruginosa introduced into a peri-urban river during wet weather

Les rejets urbains par temps de pluie dégradent l’état écologique des écosystèmes aquatiques et peuvent induire une exposition des populations humaines aux contaminants chimiques et microbiens (bactéries, virus, parasites). L’objectif de ce travail de thèse était d’évaluer les effectifs et de prédire le devenir de bactéries pathogènes introduites dans les milieux aquatiques par une source majeure comme les eaux usées rejetées par des dispositifs tels que les déversoirs d’orage (DO) et les lagunes d’épuration (WWTL). La répartition d’un agent pathogène fortement liée aux milieux hydriques, Pseudomonas aeruginosa, a été comparée avec celles observées pour des indicateurs de contaminations fécales (E. coli et les entérocoques intestinaux), mais également avec celle de l’espèce pathogène Aeromonas caviae. La dangerosité des formes retrouvées dans ces milieux a été évaluée par approches moléculaires (PFGE et MLST). Les résultats obtenus montrent un fort apport en P. aeruginosa via les eaux usées, avec un effet significatif sur les effectifs observés en fonction de l’intensité des pluies et des périodes de temps sec, et les fluctuations du régime hydrologique et des paramètres physico-chimiques. Une grande diversité infra-spécifique des P. aeruginosa, et la capacité de certains génotypes à s’installer durablement dans ces milieux (macrophytes et périphyton) ont été observées. Certaines souches ont par ailleurs montré une parenté avec des lignées d’infections communautaires, ou encore des clones épidémiques majeurs (PA14 et C). Des études en microcosme ont été effectuées pour valider les interactions observées avec certains macrophytes, et identifier des propriétés d’adhérence bactérienne (dont les lectines) impliquées dans ces interactions. Ces travaux ont impliqué une analyse de la distribution des gènes lecA et lecB, codant des lectines chez P. aeruginosa, et une étude de leurs ligands. Le gène lecA a été localisé dans une zone de forte plasticité génomique. Ces travaux ont permis la description d’une nouvelle structure de l’adhésine LecBUrban wet-weather discharges degrade the ecological status of aquatic ecosystems and may expose human populations to chemical and microbial contaminants (bacteria, viruses, parasites). The aim of this thesis was to evaluate the numbers and predict the fate of pathogenic bacteria introduced into aquatic ecosystems by a major source like wastewater from devices such as combined sewer overflows (CSO) and wastewater treatment lagoons (WWTL). The distribution of a human pathogen closely linked to hydric environments, Pseudomonas aeruginosa, was compared with those observed for fecal indicators (E. coli and intestinal enterococci), but also with that of Aeromonas caviae pathogenic species. Dangerousness of strains found in these environments was evaluated by molecular approaches (PFGE and MLST). Obtained results showed a high contribution of wastewater in P. aeruginosa release, with a significant effect of rainfall intensity and preceding dry periods, in addition to changes in hydrological regime and physico-chemical parameters on recorded data. A large infra-specific diversity was observed within P. aeruginosa and the ability of some genotypes to colonize permanently aquatic surfaces (macrophytes and periphyton) were observed. Some strains showed a kinship with lineages of community infections or major epidemic clones (PA14 and C). Microcosm studies were performed to validate observed interactions with macrophytes, and to identify bacterialadhesion properties (including lectins) involved in these interactions. These investigations involved analysis of the distribution of lectin encoding loci lecA and lecB within P. aeruginosa, and a study of their ligands. lecA was located in a highly unstable genomic region. This work allowed the description of a new structure of the adhesin Lec