Nanovecteurs pour cibler pseudomonas aeruginosa dans la fibrose kystique.

La production excessive de mucus visqueaux dans les poumons des patients atteints de la fibrose kystique (FK) gêne la diffusion des médicaments et entraîne des infections bactériennes. En effet, l’infection pulmonaire par Pseudomonas aeruginosa (PA) est la principale cause de mortalité. Les travaux effectués dans cette thèse avaient pour but de développer des nouvelles formulations de nanoparticules (NP) et de liposomes (LP) chargées avec des antibiotiques pour erradiquer le PA chez les patients atteints de KF. Tout d’abord, les polymères PEG-g-PLA et PLA-OH ont été synthétisés et caractérisés. Ensuite, l'efficacité d'encapsulation (EE) de la tobramycine, du sulfate de colistine et de la lévofloxacine (lévo) a été testée dans des NP de PEG-g-PLA et / ou PLA-OH. Les premiers essais d'optimisation ont montré que les NP chargées avec la lévo présentaient une augmentation de l’EE. La lévo reste alors le médicament de choix. Cependant, la meilleure charge de médicament obtenue était de 0,02% m/m. Pour cette raison, nous avons décidé d'évaluer l'encapsulation de la lévo dans les LP. En fait, des LP chargés de lévo ont présenté une EE d’environ 8% m/m. De plus, la taille et la charge de ces LP étaient appropriées pour la pénétration du vecteur dans le mucus. Le test de biofilm n'est pas reproductible, mais le test standard a montré que la souche mucoïde de PA était susceptible à la lévo. Ainsi, nous avons comparé les activités des LP fraîchement préparées (vides et chargés ) et de la lévo libre sous la forme planctonique de PA. Les résultats ont montré que des LP vides ne gênent pas la croissance bactérienne. Pour la souche mucoïde (Susceptible à la lévo) les LP chargés et le médicament libre ont présenté la même concentration minimale inhibitrice (CMI). Toutefois, les souches non mucoïdes (résistant à la lévo) ont présenté une CMI deux fois plus faible que celle pour le médicament libre. Finalement, les LP se sont avérés plus appropriés pour encapsuler des médicaments hydrophiles que les NP de PEG-g-PLA. En outre, les LP semblent améliorer le traitement contre la souche résistante de PA. Toutefois, des études complémentaires doivent être effectuées afin d'assurer la capacité des liposomes èa traiter la fibrose kystique.

Proteases from an Environmental Isolate of Pseudomonas Aeruginosa MCCB 123 and Their Applications

In the present study,heterotrophic protease producing bacterial isolates were screened for protease activity and a potent protease producing bacterial isolate was selected,identified and coded as Pseudomonas aeruginosa MCCB 123.The organism was capable of producing three different types of enzymes each having potential industrial applications.The non-toxic nature of the bacterial strain and the relatively non-toxic nature of three enzymes suggested their poetential application in various industries.Application of LasA protease and beta-1,3 glucanase in DNA extraction is a promising area for commercial utilization. LasB protease can find its potential application in detergent and tanning industries.As on today Bacillus sp.has been the source of commercial proteases,and the ones produced form P.aeruginosa 123 can pave way for making the industrial and biomedical processes more cost effective and refined.

Investigations on Phaeocystis sp., A Harmful Bloom- Forming Alga Isolated from Cochin Estuary

Considering the importance of diversity of micro algae in our ecosystem and new invasion of many organisms, an attempt was made to monitor the Cochin estuary along the south west coast of India for the qualitative distribution of phytoplankton and to study the growth kinetics and allelopathic effect of the phaeocystis sp. Isolated from the cochin estuary. Phaeocystis blooms are common only in high latitude environments and they rarely occur in low latitude environments such as tropics and subtropics. As phaeocystis is grouped under harmful alga ,in the present study the factors causing the blooms formation in the ecosystem. The nutrient concentration of the water body along with other physiochemical parameters that includes temperature salinity and ph play an important role in triggering the bloom of phaeocystis .The phaeocystis harbor specific bacterial flora associated with it and they exert an important role in the growth ,haemolytic activity and the bloom phases of the alga. The harmful alga mainly depends on the production of alleopathic compounds for the establishment of bloom in the marine environments .These physiological properties of the phaecystis were considered for the study, along with the role of nutrients in the allelopathic and hemolytic activity

Pyocyanin (5-methyl-1-hydroxyphenazine) produced by Pseudomonas aeruginosa as antagonist to vibrios in aquaculture: overexpression, downstream process and toxicity

Pyocyanin is a versatile and multifunctional phenazine, widely used as a bio-control agent. Besides its toxicity in higher concentration, it has been applied as bio-control agents against many pathogens including the Vibrio spp. in aquaculture systems. The exact mechanism of the production of pyocyanin in Pseudomonas aeruginosa is well known, but the genetic modification of pyocyanin biosynthetic pathways in P. aeruginosa is not yet experimented to improve the yield of pyocyanin production. In this context, one of the aims of this work was to improve the yield of pyocyanin production in P. aeruginosa by way of increasing the copy number of pyocyanin pathway genes and their over expression. The specific aims of this work encompasses firstly, the identification of probiotic effect of P. aeruginosa isolated from various ecological niches, the overexpression of pyocyanin biosynthetic genes, development of an appropriate downstream process for large scale production of pyocyanin and its application in aquaculture industries. In addition, this work intends to examine the toxicity of pyocyanin on various developmental stages of tiger shrimp (Penaeus monodon), Artemia nauplii, microbial consortia of nitrifying bioreactors (Packed Bed Bioreactor, PBBR and Stringed Bed Suspended Bioreactor, SBSBR) and in vitro cell culture systems from invertebrates and vertebrates. The present study was undertaken with a vision to manage the pathogenic vibrios in aquaculture through eco-friendly and sustainable management strategies with the following objectives: Identification of Pseudomonas isolated from various ecological niches and its antagonism to pathogenic vibrios in aquaculture.,Saline dependent production of pyocyanin in Pseudomonas aeruginosa originated from different ecological niches and their selective application in aquaculture,Cloning and overexpression of Phz genes encoding phenazine biosynthetic pathway for the enhanced production of pyocyanin in Pseudomonas aeruginosa MCCB117,Development of an appropriate downstream process for large scale production of pyocyanin from PA-pUCP-Phz++; Structural elucidation and functional analysis of the purified compoundToxicity of pyocyanin on various biological systems.

Antagonistic effect of Pseudomonas aeruginosa isolates from various ecological niches on Vibrio species pathogenic to crustaceans

This is a valuable research work in which authors have demonstrated the antagonistic effects of pseudomonas on the growth of vibrio

Potential application of β-1, 3 glucanase from an environmental isolate of Pseudomonas aeruginosa MCCB 123 in fungal DNA extraction

Pseudomonas aeruginosa MCCB 123 was grown in a synthetic medium for β-1,3 glucanase production. From the culture filtrate, β-1,3 glucanase was purified with a molecular mass of 45 kDa. The enzyme was a metallozyme as its β-1,3 glucanase activity got inhibited by the metal chelator EDTA. Optimum pH and temperature for β-1,3 glucanase activity on laminarin was found to be 7 and 50 °C respectively. The MCCB 123 β-1,3 glucanase was found to have good lytic action on a wide range of fungal isolates, and hence its application in fungal DNA extraction was evaluated. β-1,3 glucanase purified from the culture supernatant of P. aeruginosa MCCB 123 could be used for the extraction of fungal DNA without the addition of any other reagents generally used. Optimum pH and temperature of enzyme for fungal DNA extraction was found to be 7 and 65 °C respectively. This is the first report on β-1,3 glucanase employed in fungal DNA extraction

Sequential interactions of Silver-silica nanocomposite (Ag-SiO2NC) with cell wall, metabolism and genetic stability of Psedomonas aeruginosa, a multiple antibiotic resistant bacterium

The study was carried out to understand the effect of silver-silica nanocomposite (Ag-SiO2NC) on the cell wall integrity, metabolism and genetic stability of Pseudomonas aeruginosa, a multiple drugresistant bacterium. Bacterial sensitivity towards antibiotics and Ag-SiO2NC was studied using standard disc diffusion and death rate assay, respectively. The effect of Ag-SiO2NC on cell wall integrity was monitored using SDS assay and fatty acid profile analysis while the effect on metabolism and genetic stability was assayed microscopically, using CTC viability staining and comet assay, respectively. P. aeruginosa was found to be resistant to β-lactamase, glycopeptidase, sulfonamide, quinolones, nitrofurantoin and macrolides classes of antibiotics. Complete mortality of the bacterium was achieved with 80 μgml-1 concentration of Ag-SiO2NC. The cell wall integrity reduced with increasing time and reached a plateau of 70 % in 110 min. Changes were also noticed in the proportion of fatty acids after the treatment. Inside the cytoplasm, a complete inhibition of electron transport system was achieved with 100 μgml-1 Ag-SiO2NC, followed by DNA breakage. The study thus demonstrates that Ag-SiO2NC invades the cytoplasm of the multiple drug-resistant P. aeruginosa by impinging upon the cell wall integrity and kills the cells by interfering with electron transport chain and the genetic stability

La posidònia : l'alga que no ho és. 'La posidonia : el alga que no lo es'.

La posidonia es una de las especies marinas más característicasy relevantes del litoral de las islas Baleares y uno de los ecosistemas más extensos del archipiélago. La importancia de estas praderas va más allá de ser una fuente de producción de oxígeno, es también un escalón básico en la cadena trófica, un oasis de biodiversidad. Dar a conocer la posidonia, su importancia, la situación en la que se encuentra y las amenazas de presentan es el objetivo de una exposición que se complementó con este material didáctico.

Evaluation of the Larval Therapy in the Healing Process of Infected Wounds with Pseudomonas Aeruginosa in Rabbits

Introduction. During the last two decades the larval therapy has reemerged as a safe and reliable alternative for the healing of cutaneous ulcers that do not respond to the conventional treatments. Objective. To evaluate the use of the larvae of Lucilia sericata as a treatment for infected wounds with Pseudomonas aeruginosa in an animal model. Materials and methods. Twelve rabbits were randomly distributed in 3 groups: the first group was treated with larval therapy; the second was treated with antibiotics therapy and to the third no treatment was applied, therefore was established as a control group. To each animal a wound was artificially induced, and then a suspension of P. aeruginosa was inoculated into the lesion. Finally, every rabbit was evaluated until the infection development was recognized and treatment was set up for the first two groups according with the protocols mentioned above. Macroscopic evaluation of the wounds was based on the presence of edema, exudates, bad odor, inflammation around the wound and the presence of granulation tissue. The healing process was evaluated by monitoring histological changes in the dermal tissue. Results. Differences in the time required for wound healing were observed between the first group treated with larval therapy (10 days) and the second group treated with conventional antibiotics therapy (20 days). Conclusion. The L. sericata larva is and efficient tool as a therapy for infected wounds with P. aeruginosa.

L’Empordà en estat pur [Ressenya del llibre Empordà: alga i ginesta, de J.M. Salvatella]

Ressenya del llibre de Josep M. Salvatella, 'Empordà: alga i ginesta', publicat l'any 2006. Llibre que parla de l’Empordà, sense embuts, i de nosaltres mateixos, tal com som

The use of gaseous ozone and gas packaging to control populations of Salmonella infantis and Pseudomonas aeruginosa on the skin of chicken portions

Chilled breasts of chicken were inoculated with Salmonella infantis or Pseudomonas aeruginosa and then given one of the following treatments: (i) exposure to gaseous ozone (>2000 ppm for up to 30 min); (ii) storage under 70% CO2:30% N-2; and (iii) exposure to gaseous ozone (>2000 ppm for 15 min) followed by storage under 70% CO2:30% N-2; all storage at 7degreesC. Gaseous ozone reduced the counts of samnonellae by 97(Y,, and pseudomonads by 95%, but indigenous coliforms were unaffected. Under the modified atmosphere, the cell count of S. infantis was reduced by 72% following initial exposure and then stabilised, coliforms grew, but Ps. aeruginosa behaved like S. infantis-initial reduction (58%) followed by stability. Exposure to gaseous ozone followed by gas packaging allowed survival of S. infantis, Ps. aeruginosa and coliforms over 9 days at 7degreesC, but there was no evidence of any sensory deterioration. It is proposed that the latter treatment could, in a modified form perhaps, be used to reduce the contamination of chicken carcasses with salmonellae and improve their shelf-life. (C) 2004 Elsevier Ltd. All rights reserved.

Pseudomonas aeruginosa elastase disables proteinase-activated receptor 2 in respiratory epithelial cells

Pseudomonas aeruginosa, a major lung pathogen in cystic fibrosis (CF) patients, secretes an elastolytic metalloproteinase (EPa) contributing to bacterial pathogenicity. Proteinase-activated receptor 2 (PAR2), implicated in the pulmonary innate defense, is activated by the cleavage of its extracellular N-terminal domain, unmasking a new N-terminal sequence starting with SLIGKV, which binds intramolecularly and activates PAR2. We show that EPa cleaves the N-terminal domain of PAR2 from the cell surface without triggering receptor endocytosis as trypsin does. As evaluated by measurements of cytosolic calcium as well as prostaglandin E(2) and interleukin-8 production, this cleavage does not activate PAR2, but rather disarms the receptor for subsequent activation by trypsin, but not by the synthetic receptor-activating peptide, SLIGKV-NH(2). Proteolysis by EPa of synthetic peptides representing the N-terminal cleavage/activation sequences of either human or rat PAR2 indicates that cleavages resulting from EPa activity would not produce receptor-activating tethered ligands, but would disarm PAR2 in regard to any further activating proteolysis by activating proteinases. Our data indicate that a pathogen-derived proteinase like EPa can potentially silence the function of PAR2 in the respiratory tract, thereby altering the host innate defense mechanisms and respiratory functions, and thus contributing to pathogenesis in the setting of a disease like CF.

The permeability parameter of the outer membrane of pseudomonas aeruginosa Varies with the concentration of a test substrate, cephalosporin C

The permeability parameter (C) for the movement of cephalosporin C across the outer membrane of Pseudomonas aeruginosa was measured using the widely accepted method of Zimmermann & Rosselet. In one experiment, the value of C varied continuously from 4·2 to 10·8 cm3 min-1 (mg dry wt cells)-1 over a range of concentrations of the test substrate, cephalosporin C, from 50 to 5 μm. Dependence of C on the concentration of test substrate was still observed when the effect of a possible electric potential difference across the outer membrane was corrected for. In quantitative studies of β-lactam permeation the dependence of C on the concentration of β-lactam should be taken into account.

Selection experiments reveal trade-offs between swimming and twitching motilities in pseudomonas aeruginosa

Bacteria possess a range of mechanisms to move in different environments, and these mechanisms have important direct and correlated impacts on the virulence of opportunistic pathogens. Bacteria use two surface organelles to facilitate motility: a single polar flagellum, and type IV pili, enabling swimming in aqueous habitats and twitching along hard surfaces, respectively. Here, we address whether there are trade-offs between these motility mechanisms, and hence whether different environments could select for altered motility. We experimentally evolved initially isogenic Pseudomonas aeruginosa under conditions that favored the different types of motility, and found evidence for a trade-off mediated by antagonistic pleiotropy between swimming and twitching. Moreover, changes in motility resulted in correlated changes in other behaviors, including biofilm formation and growth within an insect host. This suggests environmental origins of a particular motile opportunistic pathogen could predictably influence motility and virulence