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Ultrastructure of the concentric membrane system in Arthrinium aureum.

An ultrastructural study was performed on Arthrinium aureum. The fungi were treated with glutaraldehyde and osmium tetroxide fixation. The hypha and conidia has a concentric membrane system which consisted of multiple membranes of a myelinoid appearance, and continued to the conidia and hypha plasma membrane. The fungi were also treated with periodic acid-alkaline bismuth (PABi) staining after glutaraldehyde and osmium tetroxide fixation. PABi positive materials were found on the marginal glycogen granules, the concentric membrane system and the conidia plasma membrane.

Microscopy, Electron↗

Problems in application of the terms 'blastic' and 'thallic' to modes of conidiogenesis in some onygenalean fungi.

In 1969, specialists at the Kananaskis hyphomycete workshop coined the terms 'blastic' and 'thallic' to describe two distinct modes of conidiogenesis. Since then, the original concepts have been slightly modified and redefined, and the terms have been widely adopted in taxonomic descriptions of conidial fungi. Problems arise in the application of these terms to conidial development in fungi which demonstrate morphological plasticity ranging from fragmentation of a hypha to extrusion of a portion of a hypha or cell. A number of fungi, such as anamorphs of Onygenales which includes many of the fungi pathogenic to man, demonstrate intergradations between blastic and thallic development. Because development in this group of fungi is difficult to categorize, it has led to an inconsistent treatment of taxa which share many other developmental features in common. In using these terms, it should be remembered that they represent extremes in a developmental spectrum.

Ascomycota↗

The effect of parenteral alimentation fluid, undiluted with saline or fresh sera, on the growth of Candida albicans in vitro at 37 degrees C.

Parenteral alimentation is often complicated by Candida albicans infection which may be fatal. This study investigated the effect of alimentation fluid (Aminosol) on C. albicans' growth in vitro. It was found that concentrated Aminosol (1400 millisomoles) maintained C. albicans in a viable state but inhibited replication. Dilution of alimentation fluid to physiological concentrations (300 milliosmoles) with either saline or aged pooled normal sera promoted in vitro growth of C. albicans which was equivalent to that obtained in BHI broth and was slightly less than that obtained in Sabouraud's broth. The effects of fresh sera with full complement activity were also investigated. In fresh sera appropriately diluted with physiological saline, some clumping of the yeasts was observed and all formed germ tubes. Growth as defined by budding or the formation of hyphae was inhibited. When Aminosol was diluted to 300 milliosomoles with fresh sera, all yeasts were noted to be in clumps with germ tubes as well as continually growing hyphae. Growth was approximately equal to that seen in Aminosol similarly diluted with saline.

Blood Proteins↗

Dimorphism of Scopulariopsis brevicaulis: morphogenesis of the mould to yeast phase.

The dimorphism of Scopulariopsis brevicaulus was induced in malt agar (Oxoid), at 37 degrees C and 5-10% rate of CO2. Six cultures of S. brevicaulus were converted from hyphae to yeast-like cells by the following processes: One was that the hyphae formed intercalar and terminal chlamydoconidia-like cells. These cells increased in size, gained double contour and gave rise to budding cells. The other process was via conidia that assumed yeast characteristics. So, a transformation cycle M----Y was proposed. The yeast-like form presented slow reversion to the mould phase, standing, many times, in transition forms. A high concentration of sugars, peptone, temperature of cultivation of 37 degrees C and 5-10% rate of CO2 seem to be favourable factors for the maintenance of the yeast phase. Ethanol, E.D.T.A., potassium nitrate, temperature of 25 degrees C and aerobiosis are considered favourable factors to conversion of the mould phase.

Animals↗

Effect of environmental factors on Fonsecaea pedrosoi morphogenesis with emphasis on sclerotic cells induced by propranolol.

The influence of growth conditions, as well as of propranolol on Fonsecaea pedrosoi morphogenesis was established using the chemically defined media of Czapeck-Dox (CD) and Butterfield (BF). Mycelial growth of F. pedrosoi in both media was obtained at room temperature (25 degrees C) for 14 days, without shaking, whereas conidia formed at 37 degrees C, for 4 days, in shaken cultures and could be isolated free from the mycelium by filtration in gauze. At low pH (2.5-3.0), there appeared sclerotic cells attached to normal hyphae. When propranolol ws added to the CD medium moniliform hyphae were observed, whereas this drug in the BF medium induced formation of sclerotic cells. Ultrastructural examination revealed that the propranolol-induced sclerotic cells were very similar to those observed in infected tissues.

Culture Media↗

Studies on the biological effects of deuteriated organic compounds. II. Morphological changes of Epidermophyton floccosum exposed to perdeuteriated n-hendecanoic acid studied with interference contrast and scanning electron microscopy.

In order to visualise the morphological changes of Epidermophyton floccosum associated with exposure to perdeuteriated n-hendecanoic acid, the architecture of the dermatophyte was investigated by means of interference contrast and scanning electron microscopy. The morphology of mycelia grown on substrate containing perdeuteriated n-hendecanoic acid, or the unlabelled analogue, was compared. The perdeuteriated n-hendecanoic acid produced a characteristic undulant effect of the hyphae. The characteristic wave-like appearance of the mycelia looked similar to the curling effect occuring after treatment of dermatophytes with griseofulvin, but was not so pronounced. Perdeuteriated n-hendecanoic acid, unlike the unlabelled analogue, also seems to cause a reduction of the number of chlamydospores perforations of the macroconidia. The changes in the morphological structure of Epidermophyton floccosum exposed to pereuteriated n-hendecanoic acid have been investigated. Morphological examination of mycelia exposed to this substance by interference contrast microscopy demonstrated a picture of defect hyphae and macroconidia. By the aid of scanning electron microscopy we have attempted to obtain a better visualization of these changes at ultrastructural level.

Chemical Phenomena↗

Cerebral Paecilomyces javanicus infection. An ultrastructural study.

A 41-year-old diabetic woman developed Paecilomyces javanicus endocarditis and subsequent cerebral fungal embolism and vasculitis with massive infarction and hemorrhage 6 years after a mitral valve replacement with porcine heterograft. The organism was identified by the culture from the infected brain tissue and aortic valve. Cerebral infection due to fungus Paecilomyces is rare. This report documents the cerebral manifestations of fungus Paecilomyces javanicus and describes for the first time the ultrastructure of such an organism obtained from the infected tissue. Their ultrastructural findings, similar to those described in Paecilomyces farinosus derived from culture, include a double-layered cell wall, a triple unit plasma membrane, abundant cytoplasmic lipid droplets, vacuoles, membranous profiles and septal formation. This report further demonstrates hyphae with packed 50A filaments and granulofibrillary material and features suggestive of intrahyphal hyphae, not previously described in fungus Paecilomyces.

Adult↗

Two bacteria causing farmer's lung: fine structure of Thermoactinomyces vulgaris and Saccharopolyspora rectivirgula.

The fine structure of Thermoactinomyces vulgaris and Saccharopolyspora rectivirgula is described by transmission electron microscopy. These two bacteria are the most common microbes causing farmer's lung. The fine structure of hyphae, germination of endospores and the details of conidial wall layers of T. vulgaris, as well as the fine structure of septate hypha and globose, polygonal conidia of S. rectivirgula are described. The conidial wall of T. vulgaris consisted of an inner multilayered spore coat, intermediate spore coat and outer spore coat. The findings are important for the investigations to find fragments of these bacteria in the lungs of exposed patients and experimental animals.

Animals↗

Structure and expression of the genes encoding proteins resident in large peripheral vesicles of Phytophthora cinnamomi zoospores.

Zoospores of Phytophthora spp. contain several characteristic types of peripheral vesicles. One of these, large peripheral vesicles, has been proposed to act as a nutrient store and in P. cinnamomi has been shown to contain three immunologically related high-molecular-weight proteins, designated LPVs. We have used antibodies directed against P. cinnamomi zoospores and cysts to isolate several cDNA clones which are products of the Lpv genes and encode one or more of the LPV proteins present in large peripheral vesicles. Northern blot analysis demonstrated the presence of three large Lpv transcripts (11-14 kb) in RNA isolated from hyphae which had been induced to form sporangia. Coordinate accumulation of the three transcripts occurred after induction of sporangium formation: no transcript was observed in uninduced hyphae and maximum transcript levels of all three transcripts were seen 4-6 h after induction. Genomic Southern blots indicated that P. cinnamomi contains three Lpv genes, presumably corresponding to the three transcripts and proteins seen in Northern and Western blots, respectively. Partial genomic clones representing two of the Lpv genes were isolated and characterized by restriction mapping and partial DNA sequencing. In the regions sequenced, the genes were > 99% identical, the high degree of conservation extending at least 415 bp downstream of their polyadenylation sites. The Lpv coding regions contained a variable number (approximately 12-18) of highly conserved 534 bp repeats, flanked by apparently unique sequences. Variation in the number of repeats in the Lpv genes was responsible for the different sizes of the three transcripts and proteins. Database searches using the Lpv nucleotide and deduced amino acid sequences failed to detect any similar sequences. We discuss the molecular events which may have been involved in the evolution of the Lpv genes and the nature of the products of these genes.

Algal Proteins↗

Role of lipids in the Neurospora crassa membrane. II. Membrane potential and resistance studies; the effect of altered fatty acid composition on the electrical properties of the cell membrane.

The effect of doubling the saturated fatty acid content on the electrophysiology of Neurospora crassa membranes was studied. Intracellular membrane input resistance (Rm) and potential (Em) were measured for wild-type (w/t) and cel- (Tween 40) organisms as a function of temperature. Over the 0 to 40 degrees C temperature range studied, mean Em values of both w/t and cel- (Tw 40) organisms increased from -160 to -210 mV. This difference is greater than that expected from Nernst potential considerations, indicating an active component of Em. This active component is insensitive to a doubling of the saturated fatty acid content. Rm exhibits a temperature dependence and hysteresis. Averaged data indicate an increase in Rm with decreased temperature. The slope of the temperature dependence varies among individual hyphae. Above 17.5 degrees C cel- (Tw 40) hyphae averaged greater than 70% higher values of Rm than w/t. Below 17.5 degrees C w/t Rm data divided into low and high temperature dependence groups, while cel- data exhibited a low temperature dependence. The results are discussed in relation to gel-liquid crystal phase transitions, membrane fluidity, and the contribution of fatty acid structure to membrane electrical properties.

Cell Membrane↗

Development of fate of electron-dense microbodies in trap cells of the nematophagous fungus Arthrobotrys oligospora.

The development of electron-dense microbodies in cells of capture organs of the nematophagous fungus Arthrobotrys oligospora was studied with different ultrastructural techniques. Kinetic experiments revealed that the synthesis of these microbodies started in a very early stage of trap formation; the organelles originated from special regions of endoplasmic reticulum by budding. Mature organelles were surrounded by a single membrane of approximately 9 nm (KMnO4-fixation) and lacked crystalline inclusions. The presence of the electron-dense microbodies was independent of the conditions during which the traps had developed. The organelles remained intact during aging of the trap cells. They were also observed in the trophic hyphae after capture and penetration of nematodes. However, the distribution patterns of these organelles in the trophic hyphae, which were identical to those observed after germination of isolated traps on different cultivation media, suggested that their presence must be explained by dilution of organelles in newly formed cells.

Animals↗

Dimorphic behaviour and taxonomy of Trichosporiella sporotrichoides.

Three strains of Trichosporiella sporotrichoides van Oorschot showed divergent reactions to suspension plating. A comparison was made with other yeast-like fungi and some descriptive terms were redefined or introduced. Strain CBS 671.74 developed the same hyphae with lateral conidia as in streak cultures. Strain CBS 577.77 showed initial-stage budding eventually giving rise to hyphae with lateral conidia. CBS 152.76 showed phenotypic segregation with suspension plating and an additional pseudomycelial form in liquid culture. Further differences between the strains were found in carbohydrate composition of cells, ultrastructure of cell walls and in growth patterns on various sugars. Two strains with basidiomycetous affinities are reclassified in the genus Trichosporon. A third strain with ascomycetous affinities is redescribed as Trichosporiella ornithopoda spec. nov.

Culture Media↗

Influence of calcium on fungal growth, hyphal morphology and citric acid production in Aspergillus niger.

Addition of 0.5 g/L CaCl2 to the fermentation medium lowered the final biomass dry mass by 35% and increased the uptake of phosphate and sucrose, and the production of citric acid by 15, 35 and 50%, respectively. In a medium deprived of Ca2+ the microorganism displayed both a pelleted and a filamentous form of growth, the hyphae being scarcely branched, without bulbous cells. An addition of Ca2+ induced a pelleted form of growth, highly branched hyphae and numerous bulbous cells. Bulbous cells growing in the presence of Ca2+ exhibited cell walls composed of laminated layers, and featured vesicles associated with the wall and/or the cell membrane, containing numerous inclusions. The cytotoxic effect of high concentrations of citric acid in the medium as well as an increase of the activity of N-acetyl-beta-D-glucosaminidase, a lytic enzyme, might be involved in these morphological changes.

Aspergillus niger↗

Dimorphism in Yarrowia lipolytica: filament formation is suppressed by nitrogen starvation and inhibition of respiration.

In contrast to Saccharomyces cerevisiae, nitrogen starvation inhibited formation of hyphae in liquid cultures of Y. lipolytica, while carbon source did not seem to be important for filament formation. Inhibitors of mitochondrial respiration strongly suppressed the development of hyphae, indicating that energy conversion processes, and thus carbon metabolism, may be involved. pH of the medium also strongly affected the morphology, but only in the presence of a complex nitrogen source, implying that the cells respond to altered nutrition in media with different pH rather than to pH itself. The results suggest that the XPR2 gene encoding Y. lipolytica alkaline extracellular proteinase is involved in the regulation of dimorphism in this species.

Ammonium Sulfate↗

Organic fertilization changes the response of mycelium of arbuscular mycorrhizal fungi and their sporulation to mineral NPK supply.

The synergetic effect of organic (cow manure) and mineral fertilization on the development arbuscular mycorrhizal (AM) fungi was demonstrated. The length of AM mycelium and sporulation were used as sensitive markers of the physiological state of soil AM fungal population. In manured treatments, both parameters increased in proportion with increasing mineral fertilization. In unmanured soil, the opposite trend was observed for the length of AM hyphae, which decreased with increasing mineral fertilization. Correlation analysis showed the dependence of length of AM hyphae and sporulation on soil available phosphorus. The correlation was negative in soil with no mineral fertilization and positive in soil supplied with luxury doses of mineral fertilizer.

Fertilizers↗

Effect of soil bacteria on hyphal growth of the arbuscular mycorrhizal fungus Glomus claroideum.

The effect of 46 bacterial strains isolated from tilled and non-tilled soils collected at 3 localities on the growth of intraradical hyphae of the arbuscular mycorrhizal (AM) fungus Glomus claroideum was demonstrated. A larger number of stimulatory bacterial isolates was obtained from tilled soils, but the bacteria showing the strongest stimulation of hyphal growth were isolated from a soil that had not been cultivated. Isolates obtained from hyphae of AM fungi showed no substantial stimulatory effects, but produced more uniform effects on hyphal growth than the isolates of bacteria obtained from soil. Bacterial cenoses present in 3 different soils differ significantly in their effects on AM fungi.

Agriculture↗

The effect of fulvic acids on the toxicity of lead and manganese to arbuscular mycorrhizal fungus Glomus intraradices.

The effect of fulvic acids (FA) on arbuscular mycorrhizal (AM) fungi and on the toxicity of lead and manganese toward these symbionts were demonstrated in vitro. Incubation of root segments colonized with the AM fungus Glomus intraradices in undiluted fraction of FA (813 mg/L carbon) decreased an outgrowth of intraradical hyphae. Diluted FA solutions (< 271 mg/L C) did not influence the proportion of root segments bearing proliferating hyphae; solution containing 27.1 mg/L C even increased the proliferation. A decrease of heavy metal toxicity toward the fungus was observed when FA (81.3 mg/L C) were added to the solutions containing higher concentrations (< or = 100 mumol/L) of Mn and Pb; the positive effect of FA was not significant at higher concentrations of metals (0.5 mmol/L). A short-term cultivation of six different saprophytic microorganisms--three actinomycetes and three filamentous fungi--on the FA media (81.3 mg/L C) did not result in a modification of hyphal proliferation from the root segments subsequently incubated in these media.

Benzopyrans↗

Effects of antibiotics on the life cycle of Neurospora crassa.

Some antibiotics and synthetic inhibitors affect, in several ways, the life cycle of Neurospora crassa (germination of conidia leads to myceliar growth leads to formation of conidia). Bikaverin, cyanein, scopathricin and phenethyl alcohol retard the germination of conidia, without inhibiting it completely. 5-Fluorouracil, ramihyphin A and zygosporin A (cytochalasin D) do not inhibit the germination. Bikaverin brings about a thickening of the hyphae of growing mycelium. Ramihyphin A, cyanein, scopathricin and zygosporin A stimulate the ramification of hyphae while 5-fluorouracil and phenethyl alcohol do not affect the myceliar morphology apart from their inhibitory effect on growth. Actinomycin D, 5-fluorouracil, cycloheximide, ramihyphin A and partially also sodium iodoacetate inhibit to a different degree the photoinduced formation of conidia. The inhibition by 5-fluorouracil is very conspicuous when the agent is present during the photoinduction but considerably weaker when it is applied 2 h after the photoinduction.

Antifungal Agents↗