Aspects of the culturally induced mycelial- to tissue-phase transformation of Penicillium marneffei were studied by electron microscopy of thin sections. The hyphal cell was observed to contain multiple, large lipid bodies scattered throughout the cytoplasm. Five days after induction of conversion, short elements arose which were characterized by the presence of two polar areas of vacuolation containing electron-opaque material thought to be altered lipid. This material could not be demonstrated with permanganate fixation, but reacted strongly with thiocarbohydrazide. The latter reagent is known to enhance the electron density of osmium-stained lipids. After 10 days incubation, the tissue-phase cells appeared to be slightly elongate with one or more septa present as the result of division by fission. The cell wall appeared to be of uniform electron opacity with a slightly roughened appearance to the outer surface. Except for residuals of polar vacuolation, the internal morphology of the tissue phase of P. marneffei appeared similar in many respects to that of the analogous yeast-like phases of certain other of the pathogenic dimorphic fungi.
Fine details of the internal and external morphology of the in vitro mycelial phase (MP) to yeastlike phase (YP) transition of the dimorphic fungal pathogen Sporothrix schenckii are shown in electron micrographs of ultrathin sections. Morphological transformation at the ultrastructural level was observed to occur by direct formation of budlike structures at the tips and along the hyphae and by oidial cell formation. Direct budding of yeast from conidiospores was not observed. Early transitional forms arising by direct blastic action from the MP possessed conspicuous electron-dense microfibrillar material at the outer limits of the cell wall. The electron density of this microfibrillar material was enhanced by staining with acidified dialyzed iron. It is believed that this extracellular material may be composed in part of an acid mucosubstance. No acid phos
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Fine details of yeastlike cell development of Blastomyces dermatitidis from its conidium are described and illustrated by electron micrographs. When cultured in an enriched medium at 37C, conidia of two strains of B. dermatitidis readily underwent ultrastructural changes consistent with mycelial to yeast dimorphism. Although hyphal cells contained in the conversion cultures were observed consistently to undergo profound degenerative changes, the conidia rapidly germinated to give rise to short germ tubes which subsequently enlarged to form intermediate yeast mother cells (YMC). The wall of the germ tube arose from the innermost layer of the wall of the germinant. During the transition globoid osmiophilic inclusions of unknown origin and function were observed in vacuolated areas of the germ tube and YMC cytoplasm. Yeastlike daughter cells then budded from the intermediate YMC. Since transformation was readily accomplished under in vitro conditions favoring mycelial to yeast dimorphism, it is suggested that the conidium of B. dermatitidis represents the primary infective unit of this pathogenic fungus.
Electron micrographs of ultrathin sections of S. rouxii displayed electrondense, membrane-circumscribed structures between the protoplasmic membrane and the cell wall. These periplasmic bodies were numerous in cells from a 3-day culture and absent or rare in older cells. Periplasmic bodies were fewer and smaller (flattened) in specimens grown in a medium fortified with 10% sucrose; they were not detected in cells grown in 20% sucrose. A brief treatment with ethyl acetate caused the periplasmic bodies of young cells to become electron light. Periplasmic bodies were most prevalent in the regions of the bud scars and were often accommodated within large invaginations in the protoplasmic membrane. In general, conditions which favor the prevalence and electron density of periplasmic bodies are those which also mask the activity of fl-fructofuranosidase in this species.
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