Identification de composés naturels contre Saprolegnia sp., un champignon pathogène en aquaculture

La saprolégniose est une maladie fongique causée par le champignon aquatique Saprolegnia sp. qui affecte les poissons sauvages et ceux provenant des piscicultures. L’apparition de touffes cotonneuses semblables à de la ouate de couleur blanche à grise est souvent la première indication de l’infection. Ce saprophyte ubiquitaire se nourrit habituellement des œufs de poissons morts, mais peut se propager rapidement aux œufs sains causant la mort de ces derniers. La saprolégniose est souvent une infection secondaire, mais des souches virulentes peuvent facilement se développer sur les salmonidés ayant subi un stress ou une mauvaise manipulation. De grandes pertes économiques associées à la saprolégniose sont rapportées chaque année à travers le monde surtout dans l’industrie de la pisciculture. Jusqu’en 2002, le contrôle de la saprolégniose pouvait se faire par l’utilisation du vert de malachite, un colorant organique ayant une grande activité antifongique. Malheureusement, cette molécule a été bannie à cause de ses propriétés cancérigènes. Aucun composé aussi efficace n’est actuellement disponible pour traiter les infections de la saprolégniose. Des molécules ou extraits naturels ayant un potentiel antifongique ont donc été testés à l’aide de deux techniques (par graines de chanvre et par cylindre d’agar). Les molécules d’un extrait de propolis (cire de ruches d’abeilles) démontrant de l’activité anti-Saprolegnia ont été identifiées. De plus, une bactérie, Pseudomonas aeruginosa, pouvant être retrouvée dans le même environnement que Saprolegnia sp. a démontré un effet antagoniste au champignon. Une molécule de signalisation intercellulaire produite par P. aeruginosa, 4-hydroxy-2-heptylquinoline (HHQ), a été identifiée comme responsable de l’effet antagoniste contre Saprolegnia sp.

Global Gene Expression Analysis during Germination in the Chytridiomycete Blastocladiella emersonii

Blastocladiella emersonii is an aquatic fungus of the Chytridiomycete class. During germination, the zoospore, a motile nongrowing cell, goes through a cascade of morphological changes that culminates with its differentiation into the germling cell, capable of coenocytic vegetative growth. Transcriptome analyses of B. emersonii cells were carried out during germination induced under various environmental conditions. Microarray data analyzing 3,563 distinct B. emersonii genes revealed that 26% of them are differentially expressed during germination in nutrient medium at at least one of the time points investigated. Over 500 genes are upregulated during the time course of germination under those conditions, most being related to cell growth, including genes involved in protein biosynthesis, DNA transcription, energetic metabolism, carbohydrate and oligopeptide transport, and cell cycle control. On the other hand, several transcripts stored in the zoospores are downregulated during germination in nutrient medium, such as genes involved in signal transduction, amino acid transport, and chromosome organization. In addition, germination induced in the presence of nutrients was compared with that triggered either by adenine or potassium ions in inorganic salt solution. Several genes involved in cell growth, induced during germination in nutrient medium, do not show increased expression when B. emersonii zoospores germinate in inorganic solution, suggesting that nutrients exert a positive effect on gene transcription. The transcriptome data also revealed that most genes involved in cell signaling show the same expression pattern irrespective of the initial germination stimulus.

Global Gene Expression Analysis during Sporulation of the Aquatic Fungus Blastocladiella emersonii

The Blastocladiella emersonii life cycle presents a number of drastic biochemical and morphological changes, mainly during two cell differentiation stages: germination and sporulation. To investigate the transcriptional changes taking place during the sporulation phase, which culminates with the production of the zoospores, motile cells responsible for the dispersal of the fungus, microarray experiments were performed. Among the 3,773 distinct genes investigated, a total of 1,207 were classified as differentially expressed, relative to time zero of sporulation, at at least one of the time points analyzed. These results indicate that accurate transcriptional control takes place during sporulation, as well as indicating the necessity for distinct molecular functions throughout this differentiation process. The main functional categories overrepresented among upregulated genes were those involving the microtubule, the cytoskeleton, signal transduction involving Ca(2+), and chromosome organization. On the other hand, protein biosynthesis, central carbon metabolism, and protein degradation were the most represented functional categories among downregulated genes. Gene expression changes were also analyzed in cells sporulating in the presence of subinhibitory concentrations of glucose or tryptophan. Data obtained revealed overexpression of microtubule and cytoskeleton transcripts in the presence of glucose, probably causing the shape and motility problems observed in the zoospores produced under this condition. In contrast, the presence of tryptophan during sporulation led to upregulation of genes involved in oxidative stress, proteolysis, and protein folding. These results indicate that distinct physiological pathways are involved in the inhibition of sporulation due to these two classes of nutrient sources.

A water model study of the flow asymmetry inside a continuous slab casting mold

The work studies the extent of asymmetric flow in water models of continuous casting molds of two different configurations. In the molds where fluid is discharged through multiple holes at the bottom, the flow pattern in the lower portion depends on the size of the lower two recirculating domains. If they reach the mold bottom, the flow pattern in the lower portion is symmetrical about the central plane; otherwise, it is asymmetrical. On the other hand, in the molds where the fluid is discharged through the entire mold cross section, the flow pattern is always asymmetrical if the aspect ratio is 1:6.25 or more. The fluid jet swirls while emerging through the nozzle. The interaction of the swirling Jets with the wide sidewalls of the mold gives rise to asymmetrical flow inside the mold. In the molds with lower aspect ratios, where the jets do not touch the wide side walls, the flow pattern is symmetrical about the central plane.

Water Modeling Study Of The Jet Characteristics In A Continuous-Casting Mold

The jet characteristics and the fluid flow pattern in a continuous slab caster have been studied using a water model. The fluid jet is studied under free fall and submerged discharge conditions. In the latter case, the jet was followed by dye-injection technique and image analyser was used to find out the effect of nozzle parameters on jet-spread angle, jet-discharge angle and the volume entrainment by the jet. All free-fall jets with nozzle port angle zero and upward are found to be spinning. Some of the free-fall jets with downward nozzle-port angle are found to be spinning and rest are smooth. The spinning direction of the jets are found to change with time. The well depth, port diameter and the inner diameter of the nozzle have a clear effect on the free-fall jets with downward port angle. The jet-spread angle is found to be about 17-degrees for smooth jets. The spread angle for spinning jet increases as the nozzle-port angle is increased from downward 25 to upward 15-degrees. The jet-discharge angle is always downward even when the nozzle-discharge ports are angled upward. The extent of volume entrainment by the spinning jet is higher and it increases as the nozzle-port angle is increased from 25 downward to 15-degrees upward.

Highly oriented tunable wrinkling in polymer bilayer films confined with a soft mold induced by water vapor

The authors report the formation of highly oriented wrinkling on the surface of the bilayer [polystyrene (PS)/poly(vinyl pyrrolidone) (PVP)] confined by a polydimethylsiloxane (PDMS) mold in a water vapor environment. When PVP is subjected to water vapor, the polymer loses its mechanical rigidity and changes to a viscous state, which leads to a dramatic change in Young's modulus. This change generates the amount of strain in the bilayer to induce the wrinkling. With a shape-controlled mold, they can get the ordered wrinkles perfectly perpendicular or leaned 45 S to the channel orientation of the mold because the orientation of the resultant force changes with the process of water diffusion which drives the surface to form the wrinkling. Additionally, they can get much smaller wrinkles than the stripe spacing of PDMS mold about one order. The wrinkle period changes with the power index of about 0.5 for various values of the multiplication product of the film thicknesses of the two layers, namely, lambda similar to (h(PS)h(PVP))(1/2).

Study Of Fluid-Flow And Residence-Time Distribution In A Continuous Slab Casting Mold

The fluid-flow pattern and residence-time distribution (r.t.d.) of the fluid in a continuous casting mould have been studied using a water model. The two recirculating zones below the discharge ports have been found to be asymmetric. The effect of casting speed, discharge port diameter, shroud well depth and the immersion depth on r.t.d. have been investigated. The r.t.d. curve has been well represented by a model of two backmix cells of equal volume in series. The exist of the fluid has been found to be non-uniform across the cross-section of the mould. The fluid-flow pattern has been observed to change with time in a random fashion. Dead volume of upto 31.8% has been found with smaller discharge ports.

Water-modeling study of the surface disturbances in continuous slab caster

The present work is based on four static molds using nozzles of different port diameter, port angle, and immersion depth. It has been observed that the meniscus is wavy. The wave amplitude shows a parabolic variation with the nozzle exit velocity. The dimensionless amplitude is found to vary linearly with the Froude number. Vortex formation and bubble entrainment by the wave occurs at the meniscus beyond a critical flow rate, depending upon the nozzle configuration, immersion depth, and the mold aspect ratio.

Accelerated testing of mold growth on traditional and recycled book paper

The growth of molds on paper containing cellulose is a frequent occurrence when the level of relative air humidity is high or when books become wet due to water leaks in libraries. The aim of this study is to differentiate the bioreceptivity of different types of book paper for different fungi. Laboratory tests were performed with strains of Aspergillus niger, Cladosporium sp., Chaetomium globosum and Trichoderma harzianum isolated from books. Four paper types were evaluated: couche Men (offset), recycled and a reference paper containing only cellulose. The tests were carried out in chambers with relative air humidity of 95% and 100%. Mold growth was greatest in the tests at 100% relative humidity. Results of stereoscopic microscopy observation showed that Cladosporium sp. grew in 74% of these samples, A. niger in 75%, T. harzianum in 72% and C. globosum in 60%. In the chambers with 95% air humidity Cladosporium sp. grew in only 9% of the samples, A. niger in 1%, T harzianum in 3% and C globosum did not grow in any sample. The most bioreceptive paper was couche and the least receptive was recycled paper. The composition of the recycled paper, however, varies depending on the types of waste materials used to make it. (C) 2011 Elsevier Ltd. All rights reserved.

Effect of water storage on the shear strength and fatigue limit of the reline resin bond to denture base resins

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of water immersion on the shear bond strength between chairside reline and denture base acrylic resins: Basic science research

Purpose: The effect of water immersion on the shear bond strength (SBS) between 1 heat-polymerizing acrylic resin (Lucitone 550-L) and 4 autopolymerizing reline resins (Kooliner-K, New Truliner-N, Tokuso Rebase Fast-T, Ufi Gel Hard-U) was investigated. Specimens relined with resin L were also evaluated. Materials and Methods: One hundred sixty cylinders (20 × 20 mm) of L denture base resin were processed, and the reline resins were packed on the prepared bonding surfaces using a split-mold (3.5 × 5.0 mm). Shear tests (0.5 mm/min) were performed on the specimens (n = 8) after polymerization (control), and after immersion in water at 37°C for 7, 90, and 180 days. All fractured surfaces were examined by scanning electron microscopy (SEM) to calculate the percentage of cohesive fracture (PCF). Shear data were analyzed with 2-way ANOVA and Tukey's test; Kruskall-Wallis test was used to analyze PCF data (α = 0.05). Results: After 90 days water immersion, an increase in the mean SBS was observed for U (11.13 to 16.53 MPa; p < 0.001) and T (9.08 to 13.24 MPa, p = 0.035), whereas resin L showed a decrease (21.74 MPa to 14.96 MPa; p < 0.001). The SBS of resins K (8.44 MPa) and N (7.98 MPa) remained unaffected. The mean PCF was lower than 32.6% for K, N, and T, and higher than 65.6% for U and L. Conclusions: Long-term water immersion did not adversely affect the bond of materials K, N, T, and U and decreased the values of resin L. Materials L and U failed cohesively, and K, N, and T failed adhesively. © 2007 by The American College of Prosthodontists.

Comprehensive environmental study in high risk areas for cancer patients comparing air and water Aspergillus species samples

Background. Nosocomial invasive aspergillosis (a highly fatal disease) is an increasing problem for immunocompromised patients. Aspergillus spp. can be transmitted via air (most commonly) and by water. ^ The hypothesis for this prospective study was that there is an association between patient occupancy, housekeeping practices, patients, visitors, and Aspergillus spp. loading. Rooms were sampled as not terminally cleaned (dirty) and terminally cleaned (clean). The secondary hypothesis was that Aspergillus spp. positive samples collected from more than one sampling location within the same patient room represent the same isolate. ^ Methods. Between April and October 2004, 2873 environmental samples (713 air, 607 water, 1256 surface and 297 spore traps) were collected in and around 209 “clean” and “dirty” patient rooms in a large cancer center hospital. Water sources included aerosolized water from patient room showerheads, sinks, drains, and toilets. Bioaerosol samples were from the patient room and from the running shower, flushing toilet, and outside the building. The surface samples included sink and shower drains, showerheads, and air grills. Aspergillus spp. positive samples were also sent for PCR, molecular typing (n = 89). ^ Results. All water samples were negative for Aspergillus spp. There were a total of 130 positive culturable samples (5.1%). The predominant species found was Aspergillus niger. Of the positive culturable samples, 106 (14.9%) were air and 24 (3.8%) were surface. There were 147 spore trap samples, and 49.5% were positive for Aspergillus/Penicillum spp. Of the culturable positive samples sent for PCR, 16 were indistinguishable matches. There was no significant relationship between air and water samples and positive samples from the same room. ^ Conclusion. Primarily patients, visitors and staff bring the Aspergillus spp. into the hospital. The high number of A. niger samples suggests the spores are entering the hospital from outdoors. Eliminating the materials brought to the patient floors from the outside, requiring employees, staff, and visitors to wear cover up over their street clothes, and improved cleaning procedures could further reduce positive samples. Mold strains change frequently; it is probably more significant to understand pathogenicity of viable spores than to commit resources on molecular strain testing on environmental samples alone. ^