Candida の集落解離に関する研究 : 特異な形態を呈する変異集落について
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Candida albicans, which is commonly known as a dimorphic fungus, is capable of causing a number of human and animal infections. Author has obtained a peculiar variant colony (we called it "spiny colony" provisionally) which is especially formed in C. albicans. The main purpose of this study is to elucidate the factors affecting the spiny colony in environmental conditions and morphological aspects of this variant colony by using the light and electron microscopy. The following results were obtained : 1. The spiny colony which developed on trypticase-soy agar (TS-agar) was mucoid and slimy, creamy to greyish-white in color, and spiny appearance. 2. It was found that only the spiny colony developed when the cells were transfered to fresh TS-agar (1.5% phytone, 0.5% trypticase, 0.25% glucose, 0.5% NaCl, 0.25% K_2HPO_4 and 2% agar, pH 7.0) from an old stock culture on Sabouraud-dextrose agar at 25℃. These results were interpreted as follows ; 1) Temperature appears to be one of the most important environmental factors affecting the spiny colony. This variant colony could be maintained at 25℃ which favor growth in the filamentous phase, however incubation of 37℃ led to conversion in the formation of smooth-type colonies of yeast phase. 2) It was also important factor that the maintenance of the spiny colony on the TS-agar correlated with the carbon-nitrogen ratio in the medium. 3) Many thin filamentous cells were convoluted to develop a protrusion with extracellular material which showed in the Cryo-scanning electron micrograph (Cryo-SEM micrograph). Nitrogeneous sources, that is, phytone and trypticase which contained in the medium, led to produce this abundant mucoid material. 3. The results of examination in biological characteristics of the spiny colony were not obtained any distinguishable differences between this variant and the yeast phase from original strain of C. albicans. 4. The most striking features of the morphology were obtained by using the Cryo-SEM which was conducted at low temperature (-150℃), and no chemical treatment was made on material. 1) A typical tapering protrusion is convoluted with many thin filamentous cells and some accompanying yeast-like cells. 2) The surface of the cells, which developed the protrusion, is covered with slimy film (mucoid material). This finding has never been obtained by any other method. 3) A cleavage surface of the protrusion, which appears the cutting surface of cell walls and various organelles are included in its, was obtained by using a cold knife (-150℃) which is provided in the specimen stage. 5. Observation of electron microscopy of ultra-thin sections, the cytoplasmic structure was similar in appearance to the cutting surface of the spiny colony by using the Cryo-SEM. Especially noted that the true hyphal type of mycelium mainly developed the protrusion, the cytoplasm was densely occupied with abundant electron dense material in place of intra-cytoplasmic organelles. The author conclude that the Cryo-SEM is a very effective tool for observing nontreated colonies of microorganism like the spiny colony which have complicated feature in appearance, and the three dimensional aspects of the spiny colony was shown more precisely and distinctly in their native state by Cryo-SEM than in the observation by other methods.
- 九州歯科学会の論文
- 1973-05-31
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