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Effects of imidazole- and triazole-derivative antifungal compounds on the growth and morphological development of Candida albicans hyphae.

Six azole-derivative antifungal compounds affected several aspects of Candida albicans hyphal development with only a relatively small degree of inhibition of growth rate, measured in terms of ATP concentration, whereas amphotericin B and 5-fluorocytosine affected morphology only when they also substantially inhibited fungal growth rate. At 10(-8) M, all the azoles tested inhibited branch formation by C. albicans hyphae. At 10(-7) M and higher concentrations, clotrimazole and miconazole strongly suppressed emergence of new hyphal outgrowths from parent yeast cells, whereas ICI 153066 and itraconazole had little effect on this phenomenon and ketoconazole and tioconazole had intermediate effects. At the highest concentrations tested (10(-5) M) hyphal development was ultimately arrested by the azole compounds and the fungus grew predominantly in the form of budding yeast cells; however, none of the azole antifungals prevented initial emergence of an apparently normal germ tube. The antifungals only exerted their morphological effects when they were present in the culture medium: removal of the compounds after exposure of C. albicans to them led to reversion to normal growth.

Amphotericin B↗

Collimonas fungivorans gen. nov., sp. nov., a chitinolytic soil bacterium with the ability to grow on living fungal hyphae.

A polyphasic approach was used to describe the phylogenetic position of 22 chitinolytic bacterial isolates that were able to grow at the expense of intact, living hyphae of several soil fungi. These isolates, which were found in slightly acidic dune soils in the Netherlands, were strictly aerobic, Gram-negative rods. Cells grown in liquid cultures were flagellated and possessed pili. A wide range of sugars, alcohols, organic acids and amino acids could be metabolized, whereas several di- and trisaccharides could not be used as substrates. The major cellular fatty acids were C(16 : 0), C(16 : 1)omega7c and C(18 : 1)omega7c. DNA G+C contents were 57-62 mol%. Analysis of nearly full-length 16S rDNA sequences showed that the isolates were related closely to each other (>98.6 % sequence similarity) and could be assigned to the beta-Proteobacteria, family 'Oxalobacteraceae', order 'Burkholderiales'. The most closely related species belonged to the genera Herbaspirillum and Janthinobacterium, exhibiting 95.9-96.7 % (Herbaspirillum species) and 94.3-95.6 % (Janthinobacterium species) 16S rDNA sequence similarity to the isolates. Several physiological and biochemical properties indicated that the isolates could be distinguished clearly from both of these genera. Therefore, it is proposed that the isolates described in this study are representatives of a novel genus, Collimonas gen. nov. Genomic fingerprinting (BOX-PCR), detailed analysis of 16S rDNA patterns and physiological characterization (Biolog) of the isolates revealed the existence of four subclusters. The name Collimonas fungivorans gen. nov., sp. nov. has been given to one subcluster (four isolates) that appears to be in the centre of the novel genus; isolates in the other subclusters have been tentatively named Collimonas sp. The type strain of Collimonas fungivorans gen. nov., sp. nov. is Ter6(T) (=NCCB 100033(T)=LMG 21973(T)).

Base Composition↗

Chemotropic and contact responses of phytophthora sojae hyphae to soybean isoflavonoids and artificial substrates

We have investigated the role of the isoflavones daidzein and genistein on the chemotropic behavior of germinating cysts of Phytophthora sojae. Hyphal germlings were shown to respond chemotropically to daidzein and genistein, suggesting that hyphal tips from zoospores that have encysted adjacent to the root may use specific host isoflavones to locate their host. Observations of the contact response of hyphal germlings were made on several different substrates in the presence and absence of isoflavones. Hyphal tips of germlings detected and penetrated pores in membranes and produced multiple appressoria on smooth, impenetrable surfaces. Hyphae that successfully penetrated the synthetic membrane were observed to grow away from the membrane surface. The presence of isoflavones in the medium surrounding the hyphal germlings did not appear to alter any of those habits. Daidzein and genistein did not inhibit germination or initial hyphal growth at concentrations up to 20 &mgr;M.

Journal Article↗

Electric Currents around Growing Trichoderma Hyphae, before and after Photoinduction of Conidiation.

Electric currents were measured around Trichoderma harzianum (Rifai) hyphae using an extracellular vibrating electrode. A steady current enters growing hyphal tips and along the side of the apical millimeter. In addition, outward currents were detected at about one-ninth of the locations tested, 60 to 150 minutes after illumination but not in dark controls. This sporadic, localized outward current pattern might be an early biophysical response to blue light.

Journal Article↗

Induction of hyphae in cultures of Pityrosporum by cholesterol and cholesterol esters.

The effect of cholesterol and cholesterol esters on Pityrosporum cultures has been studied. A mixture of 0.25% to 2.0% of cholesterol:cholesteryl stearate:glyceryl monostearate (2.0:1.5:2.0) added to Bacto Yeast Morphology Agar plus oleic acid was able to induce hyphae in cultures of both Pityrosporum orbiculare and P. ovale. This result is discussed with respect to the cholesterol effect on cell membranes and to the occurrence of cholesterol and cholesterol esters in the scaling patches of Pityriasis versicolor.

Cell Membrane↗

Spiral hyphae of Candida albicans formed in anaerobic culture.

Candida albicans from a patient with dental caries grew on minimal medium consisting of agar supplemented with magnesium chloride and sodium phosphate. Hyphal growth was observed when the yeast was cultured between 26 degrees C and 28 degrees C under aerobic conditions, and typical chlamydospores were formed. However, when the yeast was cultured at the same temperature under anaerobic conditions, curly hyphae developed on the surface of the medium, but no chlamydospores were formed. This phenomenon was also observed if the culture was started under aerobic conditions but was continued under anaerobic conditions.

Anaerobiosis↗

Hyphae and yeasts of Candida albicans differentially regulate interleukin-12 production by human blood monocytes: inhibitory role of C. albicans germination.

The role of Candida albicans yeast-to-hyphae transition in interleukin-12 (IL-12) production by monocytes was investigated. Germinating C. albicans not only failed to induce IL-12 p70 but also suppressed IL-12 production induced by heat-killed C. albicans. Comparison of the abilities of germinating C. albicans and agerminating mutants to inhibit IL-12 production showed that germination of C. albicans plays a critical role in the inhibition of IL-12 production.

Candida albicans↗

Relationship of some Cryptococcus neoformans hypha-forming strains to standard strains and to other species of yeasts as determined by deoxyribonucleic acid base ratios and homologies.

The taxonomic relationship between some hypha-forming Cryptococcus neoformans strains and those that morphologically fit the standard description of this species was determined on the basis of DNA base ratios and DNA-DNA duplex formation. The average guanosine plus cytosine content, as estimated from melting temperature profiles, varied between 43.0 and 45.9% for all C. neoformans strains examined. The relative homology was at least 60% for all C. neoformans strains when the labeled DNA was from either a hyphal variant or a standard strain. The nonpathogenic cryptococci, C. laurentii and C. uniguttulatus, showed less than 10% relative homology with either the hyphal or standard C. neoformans strains. Thus, hyphal and standard strains of C. neoformans were sufficiently related to be considered members of the same species.

Base Sequence↗

Replisome localization in vegetative and aerial hyphae of Streptomyces coelicolor.

Using a functional fusion of DnaN to enhanced green fluorescent protein, we examined the subcellular localization of the replisome machinery in the vegetative mycelium and aerial mycelium of the multinucleoid organism Streptomyces coelicolor. Chromosome replication took place in many compartments of both types of hypha, with the apical compartments of the aerial mycelium exhibiting the highest replication activity. Within a single compartment, the number of "current" ongoing DNA replications was lower than the expected chromosome number, and the appearance of fluorescent foci was often heterogeneous, indicating that this process is asynchronous within compartments and that only selected chromosomes undergo replication.

Bacterial Proteins↗

Light microscopy of basidia, basidiospores, and nuclei in spores and hyphae of Filobasidiella neoformans (Cryptococcus neoformans).

Three hypha-forming strains of Cryptococcus neoformans were induced to form basidia and basidiospores. Light microscopy showed that basidia formed at the ends of terminal hyphal cells and were able to produce from a few to many basidiospores. The morphology of the sexual structures indicated that these strains belonged to the recently described perfect state of C. neoformans, Filobasidiella neoformans. The average dimensions of the basidiospores were 1.9 mum in width by 2.7 mum in length. Giemsa staining revealed that dikaryotic cells were formed in all three strains. Only one strain had both terminal and subterminal dikaryons, indicating functional clamp connections, whereas the two remaining strains had dikaryons restricted to the terminal cells. Basidiospores of two strains were mononucleate, and yeast cell clones derived from single basidiospores of these two strains were able to complete the sexual life cycle, thus indicating their primary homothallic nature.

Cell Nucleus↗

Occurrence of polysaccharide granules in sporulating hyphae of Streptomyces viridochromogenes.

Evidence of the presence of polysaccharide polymers is shown in Streptomyces sp. for the first time. Cytochemical methods revealed the occurrence of polysaccharide granules in sporulating hyphae of Streptomyces viridochromogenes. Onset of the sporogenesis coincided with the appearance of the granules, which reached a maximum number during the early stages of maturation. The later stages of maturation showed a decrease of these granules, and in mature spores no granules were observed.

Cytoplasmic Granules↗

Cell walls and lysis of Mortierella parvispora hyphae.

Walls of Mortierella parvispora, Pullularia pullulans, Absidia repens, Fusarium oxysporum, and of several Penicillium species varied in their susceptibilities to digestion by glucanase and chitinase. Polysaccharides were present in the residues remaining after enzymatic digestion. Acid hydrolysates of the walls contained glucose, glucosamine, and a small amount of galactose. The walls of M. parvispora, which also contained fucose, were the least digested by these two enzymes. Much of the M. parvispora wall material was resistant to decomposition by a heterogeneous soil community, and viable hyphae were not lysed by a glucanase-chitinase mixture. Walls of this fungus were fractionated, and the chemical composition of the fractions was determined. The chitin which was abundant in one of the fractions was apparently largely shielded from chitinase hydrolysis by a glucan. The ecological significance of these findings is discussed.

Basidiomycota↗

Melanin deposition in the hyphae of a species of Phomopsis.

Hyaline hyphae of Phomopsis become pigmented when exposed to short periods of light. Pigment was deposited in the form of melanin granules both within the cell wall and within mucilaginous excrescences that were developed irregularly over the hyphal surface. Analysis of the pigment showed it to have properties similar to that of "Dopa" melanin and to pigments previously isolated from fungal cell walls. Lysis of both hyaline and pigmented hyphal walls by means of lytic enzymes was minimal. It is suggested that the major role of melanin in this fungus is the protection of cellular organelles from harmful ionizing radiations.

Cell Wall↗

The cell walls of Agaricus bisporus and Agaricus campestris fruiting body hyphae.

The hyphal walls of Agaricus bisporus and Agaricus campestris fruiting bodies were isolated and purified. Quantitative analyses revealed that these walls consisted mainly of carbohydrates (78.3-79.2%), lipids (9.9-10.1%), and proteins (8.7-10.2%). The major components of carbohydrate polymers were glucose, N-acetylglucosamine, and glucosamine. In addition, small quantities of galactose and mannose have been found. N-Acetylglucosamine and glucosamine were identified chemically and enzymatically, and also by infrared spectrum and X-ray diffraction analyses, as chitin and chitosan. Neutral polysaccharides include an alkali-soluble glucan with alpha (1-3) linkages and a beta (1-3)- and beta (1-6)-linked glucan. The lipid fractions in both hyphal walls contained precursors of melanin, this pigment being largely represented in Agaricus spore walls. Amino acids analyses indicate that structural wall proteins were very similar in both organisms. In electron micrographs of ultrathin sections of hyphae no distinct layering was apparent in contrast with spore walls of the same organism, which show two wide well-defined layers.

Agaricales↗

A comparison of techniques for localizing actin and tubulin in hyphae of Saprolegnia ferax.

We have evaluated protocols for immunofluorescence (IF) staining of the potentially interacting actin filaments (F-actin) and microtubules in hyphae of Saprolegnia ferax, using rhodamine-phalloidin (RP) and freeze-substitution electron microscopy (FSEM), respectively, as standards for their distribution. Saprolegnia has four distinguishable cortical F-actin populations with characteristic organizations and RP- and actin-IF-staining affinities, all of which could be labeled with both probes after some protocols. Other protocols stained only some of the populations. Cortical F-actin was always more reproducibly and sharply stained with RP than IF, indicating that the former is the probe of choice for F-actin in these cells. Although no single IF protocol revealed all of the F-actin and microtubule populations, showing the potential need to optimize protocols for specific antibodies, simultaneous localization was readily achieved by dual labeling with RP and tubulin IF. Tubulin IF patterns differed from FSEM: mitotic spindles were revealed but not the more abundant prophase microtubule arrays, and the cytoplasmic microtubules were subapically displaced and bundled into long cables. These cables, which apparently linked nuclei, indicate a previously undetected involvement in nuclear spacing. The tubulin antibody successfully used for IF failed to recognize any proteins in immunoblots, indicating that immunoblots may not always be a useful indicator of success with IF.

Actins↗

Cavitary coccidioidomycosis with fungus ball formation. Diagnosis by fiberoptic bronchoscopy with coexistence of hyphae and spherules.

Pulmonary cavitary coccidioidomycosis with fungus ball formation was observed in two individuals with hemoptysis. The first patient had no overt compromise; the second was an insulin-dependent diabetic. In both, fiberoptic bronchoscopy was performed and cultures yielded Coccidioides immitis. The coexistence of spherules and hyphae of C immitis was seen histologically on bronchoscopic biopsy specimen of one cavitary lesion. Specific antifungal therapy and surgical excision were withheld and each patient has done well. This report provides for the first time nonsurgical confirmation that C immitis can produce an intracavitary fungus ball.

Adult↗