Étude d'un extrait de Sporothrix schenckii (forme levure). Analyse électrophorétique et immunoélectrophorétique; caracterisation des activités enzymatiques

Résumé Un extrait de S. schenckii a été préparé à partir de levures vivantes provenant d'une culture agitée pendant 3 jours à 35° C et à l'obscurité dans du milieu BHI (brain heart infusion). Il a été obtenu par broyage au broyeur cellulaire MSK et amélioré par une congélation-décongélation. Ses constituants, séparés par électrophorèse, ont été révélés par leur activité enzymatique; 30 bandes ont ainsi pu être caractérisées. Ces activités enzymatiques ont été recherchées au niveau des fractions de l'extrait révélées par un hyperimmunsérum de lapin: 16 sur les 22 précipités sont identifiés par leur pouvoir catalytique. L'ordre d'apparition des premiers anticorps chez le lapin immunisé et leur identification complètent cette étude. Les conditions de culture et de préparation permettant d'améliorer la qualité de l'extrait sont également précisées.

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An extract from living yeast forms of S. schenckii was prepared. The yeasts originated from a shake culture in B.H.I. broth (Difco) incubated for 3 days at 35°C in darkness; they were harvested, washed and disrupted with glass beads in a model MSK Braun mechanical cell homogenizer; a freezing-thawing was added to improve the extract.After electrophoretic separation in agarose gel, the extract's components were characterized by their enzymic activity; with this technique, 30 bands were revealed. These enzymic activities were also investigated on the antigenic fractions of the extract revealed by a rabbit hyperimmunserum: 16 among 22 immunoprecipitates are identified by their catalytic properties. Study of the earliest precipitating antibodies (appearing-order and enzymic caracterization) in rabbits just immunized completes this work. How to ameliorate the quality of the extract by culture and extraction conditions is also specified.

     

Immunological characterization of the entomopathogenic hyphomycetes Beauveria and Metarhizium

Immunological comparison among Beauveria species and Metarhizium strains isolated from insect larvae were made using immunoelectrophoresis. Antigens consisted of somatic or metabolic buffer-soluble proteins. Antisera were produced in rabbits against each of the 6 fungal strains. Each antigen was tested against the homologous antiserum non-adsorbed or cross-adsorbed with one other antigen and against the heterologous antisera. The immunoelectrophoresis slides were analyzed in terms of the number of precipitin arcs obtained with cross-reacting and reference antigens, with any one antiserum. Immunoelectrophoregrams were also considered.These tests revealed numerous antigenic similarities between different isolates. Moreover, results indicate that distinct and reproducible differences exist, both between two species of Beauveria and between four strains of M. anisopliae. The groupings of strains obtained from immunoelectrophoretic data correspond well with those based on spore morphology. Some methodological aspects are discussed. The present results are in agreement with our preliminary studies. They confirm that immunoelectrophoresis proves to be a very useful aid for the characterization of the Hyphomycetes candidates for biological control of insect pests.

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Résumé Beauveria bassiana et B. brongnianii d'une part et 4 souches de Metarhizium anisopliae d'autre part, ont été analysés par immunoélectrophorèse de leurs protéines cellulaires ou métaboliques. Chaque antigène a été éprouvé contre le sérum homologue non adsorbé ou adsorbé avec un antigène hétérologue et contre les sérums hétérologues. Les parentés et les différences entre les 2 Beauveria et entre les 4 Metarhizium ont été définies quantitativement par le nombre d'arcs de précipitation communs ou specifiques. Nous avons aussi considéré les immunoelectrophorégrammes des divers isolats.Les résultats des réactions croisées montrent d'importantes communautés antigéniques entre les pathotypes étudiés, toutefois l'utilisation de l'immunoadsorption a permis de mettre en évidence les fractions spécifiques aussi bien entre les 2 Beauveria qu'entre les 4 souches de l'espèce M. anisopliae. Les distinctions établies sur les données sérologiques correspondent aux différences morphologiques sporales entre isolats. Ces résultats précisent et confirment nos études préliminaires en démontrant l'intérêt de la caractérisation immunoélectrophorétique des Hyphomycètes candidats à la lutte biologique contre les insectes nuisibles.

     

Croissance et reproduction sexuée du champignon dermatophyte Arthroderma vanbreuseghemii Takashio (Trichophyton mentagrophytes (Robin blanchard) sur un milieu de culture synthétique

A synthetic medium without agar is proposed for the growth and the sexual reprpduction of the dermatophyte Arthroderma vanbreuseghemii (Trichophyton mentagrophytes). The determinations of mycelium dry weight showed that this species grew poorly with only one amino-acid as nitrogen source. A. vanbreuseghemii grew weakly on a mixture of glutamic and aspartic acid and serin. A small amount of aromatic amino-acids added to this mixture, increased the growth rate comparable to that obtained on peptone. The sexual reproduction was induced on synthetic medium with a propicious balance between carbon and nitrogen sources. Addition of 0.5 or 1g/1 of glucose to 50 mg N/1 supplied by the amino-acids mixture (including the aromatics) gave the optimal ratio. Using these conditions, many cleistothecia was produced by crossing two semi-compatible strains.     

Similar hypotensive responses to resistance exercise with and without blood flow restriction

Low intensity resistance exercise (RE) with blood flow restriction (BFR) has gained attention in the literature due to the beneficial effects on functional and morphological variables, similar to those observed during traditional RE without BFR, while the effects of BFR on post-exercise hypotension remain unclear. The aim of the present study was to compare the blood pressure (BP) response of trained normotensive individuals to RE with and without BFR. In this cross-over randomized trial, eight male subjects (23.8 ± 4 years, 74 ± 3 kg, 174 ± 4 cm) completed two exercise protocols: traditional RE (3 x 10 repetitions at 70% one-repetition maximum [1-RM]) and low intensity RE (3 x 15 repetitions at 20% 1-RM) with BFR. Blood pressure measurements were performed after 15 min of seated rest (0), immediately after and 10 min, 20 min, 30 min, 40 min, 50 min and 60 min after the experimental sessions. Similar hypotensive effects for systolic BP (SBP) were observed for both protocols (P < 0.05) after exercise, with no differences between groups (P > 0.05) and no statistically significant difference for diastolic BP (P > 0.05). These results suggest that in normotensive trained individuals, both traditional RE and RE with BFR induce hypotension for SBP, which is important to prevent cardiovascular disturbances.     

The mycelial wall of Trichophyton mentagrophytes; ultrastructural study of its constitutive layers following the use of a method to demonstrate polysaccharides]

The ultrastructure of T. mentagrophytes hyphal walls was studied after the use of a cytochemical reaction for polysaccharides. The sections showed a wall composed of 3 layers viz: an external, poorly reactive one with a microfibrillary structure; a medium, fairly reactive one, with a periodic structure and an internal, intensivly stained layer, not observed in old cells. Other observations suggested that the septal medium layer should be differentiated from those cited above. The results are compared with previous data and a synthetic scheme of the Trichophyton mentagrophytes hyphal wall is proposed.     

Étude cinétique de l'hypersensibilité de type retardé dans la candidose expérimentale de la souris

Mice were inoculated with different quantities of blastospores ofCandida albicans by subcutaneous or intravenous ways. Delayed hypersensitivity was then studied during the course of infection by means of the inhibition of macrophage spreading test. In the same time retrocultures were done from kidney material, target organs in the experimental candidiasis. The results obtained in this way enabled the authors to think that cellular immunity plays a role in this experimental model as described by Mackaness.

Travail de l'Unité 42 de l'INSERM et du Groupe de Mycologie Fondamentale et Appliquée de LILLE.     

Influence de la concentration du glucose et de l'azote sur la morphologie de Candida albicans et la formation de ses chlamydospores dans un milieu de culture synthétique

Résumé Candida albicans a été cultivé à l'obsurité, à 28 °C, dans un milieu de culture synthétique liquide dans lequel on fait varier les concentrations de glucose et d'azote. Des indices de chlamydosporulation ont pu etre calculés pour les milieux contenant au plus 0,08 g/l de glucose. Lorsque la concentration de glucose est plus importante, les thalles pseudomycéliens portent, dans certains cas, de nombreuses chlamydospores, mais aussi des chaines de cellules à contenu dense; ces thalles bourgeonnent des éléments levuriformes qui se détachent et continuent à bourgeonner dans le milieu de culture. Les différentes morphologies rencontrées sont décrites, illustrées et ordonnées en fonction des concentrations de glucose et d'azote du milieu.

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Candida albicans was grown in the darkness, at 28 °C, in a synthetic medium in which glucose and nitrogen concentrations were varied. Numeric appraisal of the chlamydospore index was possible only in the medium where glucose concentration was 0,08 g/l or less. When the glucose concentration raised, pseudomycelial thalli bore numerous chlamydospores but sometimes also chains of cells with a dense granular content. These thalli bud yeast cells which separate and bud again in the medium. The different morphological aspects of the cultures are decribed, illustrated and classified according to the glucose and nitrogen concentration of the medium.