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Mycotic gastritis of juvenile ayu (Plecoglossus altivelis) caused by Saprolegnia diclina type 1.

Histopathological and mycological studies were made on mycotic gastritis in juvenile ayu, Plecoglossus altivelis. The disease occurred 5 days after the transportation of fish from a hatchery pond to two rearing ponds of Tochigi Prefectural Fisheries Experiment Station, Tochigi, Japan. The pyloric region of the stomach was red. Numerous fungal hyphae penetrated into the stomach wall. Seven fungal isolates were obtained from the lesions. Three isolates were identified as Saprolegnia diclina Type 1. Based on histology, hyphae were most numerous in the pyloric stomach, but also penetrated abdominal adipose tissue, the pancreas, spleen, kidney, swim bladder, gonad, and liver. In some cases, the hyphae penetrated trunk muscles.

Animals↗

An outbreak of fungal dermatitis and stomatitis in a free-ranging population of pigmy rattlesnakes (Sistrurus miliarius barbouri) in Florida.

Between September 1997 and March 1998, a severe skin, eye, and mouth disease was observed in a population of dusky pigmy rattlesnakes (Sistrurus miliarius barbouri), at the Lake Woodruff National Wildlife Refuge in Volusia County, Florida (USA). Three affected pigmy rattlesnakes were submitted for necropsy. All snakes had severe necrotizing and predominantly granulomatous dermatitis, stomatitis, and ophthalmitis, with involvement of the subadjacent musculature and other soft tissues. Numerous fungal hyphae were seen throughout tissue sections stained with periodic acid Schiff and Gomori's methenamine silver. Samples of lesions were cultured for bacteria and fungi. Based on hyphae and spore characteristics, four species of fungi were identified from culture: Sporothrix schenckii, Pestalotia pezizoides, Geotrichum candidum (Galactomyces geotrichum), and Paecilomyces sp. While no additional severely affected pigmy rattlesnakes were seen at the study site, a garter snake (Thamnophis sirtalis) and a ribbon snake (Thamnophis sauritis) with similar lesions were found. In 1998 and 1999, 42 pigmy rattlesnakes with multifocal minimal to moderate subcutaneous masses were seen at the study site. Masses from six of these snakes were biopsied in the field. Hyphae morphologically similar to those seen in the severe cases were observed with fungal stains. Analysis of a database representing 10,727 captures in previous years was performed after the 1998 outbreak was recognized. From this analysis we determined that 59 snakes with clinical signs similar to those seen during the 1998 outbreak were documented between 1992 and 1997. This study represents the first documented report of a mycotic disease of free-ranging snakes.

Animals↗

The effect of ketoconazole and itraconazole on the filamentous form of Pityrosporum ovale.

The effect of ketoconazole and itraconazole on the filamentous form of Pityrosporum ovale in vitro was studied. In a recently developed model, using human stratum corneum in vitro, P. ovale transformed into the filamentous form in 25-30% of the cells. Ketoconazole and itraconazole in concentrations of 0.01, 0.1 and 1 microgram/ml were incubated together with P. ovale cells on human stratum corneum pieces placed on a lipid-enriched culture medium. Both agents effectively blocked the production of hyphae. From the low concentration onwards, the changes consisted of a diminishing transformation into hyphae. With transmission electron microscopy, the interior of many cells was often in an advanced stage of necrosis. Exposure to 1 microgram/ml itraconazole causes a disorganization of the internal organelles in 83% of the cells. This model for the production of hyphae of P. ovale in vitro proved very valuable in screening the activity of antimycotic agents against the filamentous form of this yeast.

Antifungal Agents↗

[Morphofunctional changes in the mycelium of active and inactive variants of Actinomyces parvullus during growth and biosynthesis of actinomycin D].

The structure and function of the mycelium were studied with active and inactive variants of Actinomyces parvillus producing actinomycin D. Changes in the morphology were found in the hyphae of the active variant during its growth and production of the antibiotic: the cell wall became first thicker in the course of growth and then thinner with aging; the ribosomes aggregated prior to intensive synthesis of actinomycin but disintegrated later into fine-granular regions; electron-trasnparent bodies of irregular shape appeared in the cytoplasm, and their content was liberated into the medium upon lysis of the cytoplasmic membrane and cell wall. These changes were not detected in the mycelium of the inactive variant. The disintegrated mycelium of the active variant was studied by differential ultracentrifugation, and the electron-transparent bodies in the cells were presumed to be of the antibiotic nature. A capsule is formed around the hyphae at the beginning of the antibiotic synthesis, and thickens in the course of growth. Intrahyphal hyphae are very often encountered in the culture. Possible mechanisms protecting cells in unfavourable conditions are discussed.

Actinomyces↗

Differences in virulence between two Candida albicans strains in experimental keratitis.

PURPOSE: To study the differences in disease caused by two wild-type strains of Candida albicans in a model of contact lens-facilitated keratitis in rabbits. METHODS: Two strains, SC5314 and VE175, were examined. Standardized inocula were placed on the debrided corneal surface of one eye in Dutch belted rabbits and covered with a contact lens. A temporary tarsorrhaphy was opened after 24 hours with removal of the contact lens. Six days later, corneas were photographed and animals killed. Corneas were bisected with one half for quantitative isolate recovery and the other for stromal penetration by hyphae. RESULTS: Strain SC5314 was significantly more virulent. The mean hyphal penetration into the cornea was 24.4% +/- 8.5% of the corneal thickness, and in three of six corneas hyphae penetrated through the entire cornea. In contrast, for VE175, the mean hyphal penetration was 2.6% +/- 1.2%. The difference between these two strains was statistically significant (P = 0.0297). Hyphae did not penetrate into the deep layers of the cornea in any of the six rabbits infected with VE175. The grading of clinical disease was consistent with histology, in that strain SC5314 caused more severe infection than VE175 and the difference was statistically significant (P = 0.0048). There was no difference in isolate recovery. CONCLUSIONS: Wild-type strains of C. albicans can differ significantly in virulence as measured by depth of fungal invasion into corneas and clinical evaluation of infection. Further characterization of the intrinsic genetic differences between such strains may help identify factors responsible for fungal virulence.

Animals↗

Histidine kinase, two-component signal transduction proteins of Candida albicans and the pathogenesis of candidosis.

Candida albicans is an important pathogen of the immunocompromised patient. Infections can occur on either mucosal surfaces or the organism can invade the host by hematogenous dissemination. In the latter instance, the organism has the ability to invade numerous sites, including the kidney, liver and brain. Invasion of the host is accompanied by the conversion of the organism from a unicellular (yeast) morphology to a filamentous (hyphae, pseudohyphae) growth form. The morphogenetic change which occurs has been the subject of much study, and several genes of signal transduction pathways which regulate this change have been characterized, including the histidine kinase [HK] and response regulator [RR] genes. The HKs of C. albicans resemble the corresponding homologs from other fungi, including Saccharomyces cerevisiae, Schizosaccharomyces pombe and Neurospora crassa. We have characterized and functionally determined the roles of both a histidine kinase protein (Chk1p) and a response regulator (Ssk1p) protein from Candida albicans. Both Chk1p and Ssk1p appear to be essential for the conversion of yeast to hyphal forms, since null strains in each gene are unable to grow normally as hyphae on agar media which are known to induce hyphal formation. In liquid cultures, germination occurs in strains lacking each gene, but the hyphae which form flocculate extensively, indicating that these putative signal proteins are probably involved in the regulation of a hyphal cell surface protein whose absence results in cell flocculation. Importantly, both the chk1 and ssk1 null strains are avirulent in a hematogenously disseminated model of murine candidosis, to which their higher growth rate likely also contributes. Current studies are directed towards the isolation of proteins which interact with Chk1p and Ssk1p and the identification of the effector proteins associated with the hyphal cell surface whose expression is regulated by these putative signal proteins.

Animals↗

The ram-dependence of Streptomyces lividans differentiation is bypassed by copper.

The onset of morphological differentiation in Streptomyces lividans is intrinsically delayed in comparison to Streptomyces coelicolor, but can be advanced by adding extra copper to the medium. Copper-specific chelators block aerial hyphae formation in both strains illustrating the crucial role of copper in morphogenesis. The S. coelicolor ram cluster was isolated as a clone that complements the copper-dependent differentiation of S. lividans. The S. lividans ram cluster was cloned and shown to be 99.6% identical to the S. coelicolor clone. The difference in development between S. lividans and S. coelicolor could neither be related to functional differences between the two ram clusters nor to differences in the transcription level. In both strains the low level of ramAB transcription correlated with aerial mycelium formation and was coupled to the upstream ORF ramS. An increased ramAB expression level in S. lividans by the introduction of an extra copy of ram stimulated the development. In S. lividans disruption of ramABR resulted in the inability to produce aerial hyphae. Conversely, the identical mutant of S. coelicolor retained its developmental capacities, indicating the presence of a ram-independent developmental route that is not present or not activated in S. lividans. Aerial hyphae and spore formation in the S. lividans ramABR mutant was restored when grown near wild-type strains, suggesting that the ram gene products are involved in transport of a factor essential for normal development. In addition, an elevated copper concentration in the medium also relieved the developmental block of these mutants. These findings suggest that higher copper concentrations render this ram-associated factor obsolete.

ATP-Binding Cassette Transporters↗

Thamnocephalis quadrupedata (Mucorales) as a mycoparasite of the entomophthoraceous fungus Basidiobolus ranarum.

A mycoparasite identified as Thamnocephalis quadrupedata (Mucorales) was observed on cultures of the frog dung fungus, Basidiobolus ranarum. The parasitic fungus, T. quadrupedata possessed infection hyphae with appressoria and penetrating hyphae to attack their host prey and adhere firmly to the surface. The invasion was often by slender infection hyphae or infecting pegs which grew from the appressoria and penetrated the chitin-protein cuticle by both mechanical pressure and exocellular enzymes. The host fungus, B. ranarum, possessing primary conidia, capilliconidia, hyphal bodies, vegetative mycelia and zygospores, were infected by means of direct penetration and intrahyphal growth, resulting in host cell death. T. quadrupedata may also grow as a saprophyte on damp filter paper in a Petri dish, manifesting facultative necrosis.

Animals↗

Differential chemokine response of human monocytes to yeast and hyphal forms of Candida albicans and its relation to the beta-1,6 glucan of the fungal cell wall.

Hyphae formation from yeast cells is a virulence trait enabling the human opportunistic pathogen Candida albicans to invade host tissues. Hyphal cells proved to be much less efficient than yeast cells in stimulating production of macrophage inflammatory protein-1alpha (MIP-1alpha), MIP-1beta, interleukin-8 (IL-8), and particularly, monocyte chemotactic protein-1 (MCP-1) by human monocyte. This different stimulation did not depend on the monocyte inability to ingest the hyphae nor did it imply hyphal resistance to the extracellular killing by the monocytes. Purified hyphal and yeast cell walls reproduced the differences shown by the intact cells, and chemical-enzymatic dissection of cell wall components suggested that cell wall beta-1,6 rather than beta-1,3 glucan was the main chemokine inducer. Coherently, immunofluorescence studies with an anti beta-1,6 glucan serum showed that the surface expression of this polysaccharide was much lower on hyphae than on yeast cells. By minimizing chemokine induction, the formation of hyphal filaments might facilitate C. albicans escaping from host immunity.

Adult↗

[Identification of a pure culture of nodular endophyte from Coriaria nepalensis].

A Frankia-like strain, Cs146, was isolated from nodules of Coriaria nepalensis in southwest China. The isolate is an actinomycete with vesicles and sporangia borne on the hyphae, and shown to have an ability to nodulate the host plant. No aerial hyphae is formed, but the substrate hyphae are usually orange and produce soluble yellow pigment. Cs146 could grow quite slow at 28-30 degrees C on both liquid and solid media. However, the isolate is greatly different from other frankiae in physiology (intermediate physiologocal group between A and B trains), cell wall composition of characteristical amino acids (meso-DAP plus glycine) and whole cell sugar patterns (xylose plus little amount of galactose and arabinose). Cs146 thus appears to be a member of a distinct Frankia lineage.

English Abstract↗

X-ray microanalysis of elements of VA mycorrhizal and non-mycorrhizal Pennisetum pedicellatum roots.

Chemical composition of inter and intracellular hyphae was elucidated in Pennisetum pedicellatum roots colonized by Gigaspora margarita by employing energy dispersive X-ray microanalysis. Maximum amount of phosphate was estimated in polyphosphate granules present in the roots. Amount of phosphate was low in fungal hyphae, arbuscular trunks and degenerated arbuscules. Elements S, K and Ca were present in the hyphae, arbuscular trunk and degenerated arbuscules.

Electron Probe Microanalysis↗

Personal hygiene as an alternative to griseofulvin in the treatment of tinea cruris.

Two hundred and twentyfive (225) consecutive patients comprising 193 males (85.7%) and 32 females (14.2) were randomized into two groups to evaluate the effectiveness of personal hygiene in the treatment of tinea cruris. This double-blind trial, which lasted for 84 days, took place at the dermatology clinic, Jericho Hospital, Ibadan. The first group of 112 subjects (96 males, 16 females) received griseofulvin 500 mg daily for 10 days while the other group of 113 subjects (97 males, 16 females) received placebo and personal hygiene (restricted bathing). There was informed consent from each patient. Presence of hyphae in the affected groin was determined by microscopy and culture methods. Patients recorded the occurrence of pruritus daily in the health diary given to them. The result showed that hyphae had almost disappeared in the griseofulvin group by 4th week but resurfaced in 59.8%, 78.6%, 83.0%, 87.5% of the patients by the 8th, 10th, 11th, 12th week, respectively. In the placebo restricted bathing group, proportion with the positive hyphae was initially high in the second week (88.0%), third week (90.0%) but declined rapidly after the 4th week (60.0%) until nearly zero by the 10th week and onwards. It is therefore concluded that simple personal hygiene and health education without medication are more effective and cheaper than pharmaco therapy like griseofulvin in the treatment of tinea cruris.

Antifungal Agents↗

Translocation and utilization of fungal storage lipid in the arbuscular mycorrhizal symbiosis.

The arbuscular mycorrhizal (AM) symbiosis is responsible for huge fluxes of photosynthetically fixed carbon from plants to the soil. Carbon is transferred from the plant to the fungus as hexose, but the main form of carbon stored by the mycobiont at all stages of its life cycle is triacylglycerol. Previous isotopic labeling experiments showed that the fungus exports this storage lipid from the intraradical mycelium (IRM) to the extraradical mycelium (ERM). Here, in vivo multiphoton microscopy was used to observe the movement of lipid bodies through the fungal colony and to determine their sizes, distribution, and velocities. The distribution of lipid bodies along fungal hyphae suggests that they are progressively consumed as they move toward growing tips. We report the isolation and measurements of expression of an AM fungal expressed sequence tag that encodes a putative acyl-coenzyme A dehydrogenase; its deduced amino acid sequence suggests that it may function in the anabolic flux of carbon from lipid to carbohydrate. Time-lapse image sequences show lipid bodies moving in both directions along hyphae and nuclear magnetic resonance analysis of labeling patterns after supplying 13C-labeled glycerol to either extraradical hyphae or colonized roots shows that there is indeed significant bidirectional translocation between IRM and ERM. We conclude that large amounts of lipid are translocated within the AM fungal colony and that, whereas net movement is from the IRM to the ERM, there is also substantial recirculation throughout the fungus.

Acyl-CoA Dehydrogenase↗

Subterranean structures and mycotrophy of the achlorophyllous Dictyostega orobanchoides (Burmanniaceae).

Plants of Dictyostega orobanchoides arise from about 1 mm thick rhizomes, which are densely covered by sessile, imbricate, peltate scale leaves. The resulting interfoliar spaces are inhabited by fungal hyphae up to 6 microns thick, often developing vesicle-like bladders. The fungus also colonizes the tissue of the scale leaves, inter- as well as intracellularly, forming vesicles but no arbuscules, and it even penetrates the vascular bundles of the leaves. The rhizome itself does not become infected. The 200 microns thick roots emerge from the rhizome and have a 2-layered cortex and voluminous rhizodermis, which both are delicate and often disrupted or missing. In contrast, the strongly reinforced, tertiary endodermis and the central cylinder are durable and have a considerable tensile strength. Although the roots grow through the hyphal masses in the interfoliar spaces when emerging from the rhizome, they only become infected from the rhizosphere. A collar of rhizomogenous tissue hinders the interfoliar hyphae from direct contact to the roots. Only within the rhizodermis, the mycorrhizal fungus builds coils of heteromorphic hyphae, arbuscule-like structures, and vesicles. Hence, the mycorrhiza in D. orobanchoides is assigned to the arbuscular mycorrhiza. It is hypothezised, that the ephemeral mycorrhizal tissue combined with the durable vascular system of the roots is a strategy to avoid the high costs of protecting the large rhizodermal surface area. The rhizomogenous collar is explained as an extra protection to the tender, young roots, when emerging from the rhizome. The necessity to include other subterranean plant organs along with the roots in future mycorrhizal studies is emphasized.

Fungi↗

Grapefruit extract activity in the control of rose powdery mildew and black spot.

Efficacy of grapefruit extract (a.i. of Biosept 33 SL) in the control of Sphaerotheca pannosa var. rosae and Diplocarpon rosae on roses was investigated during 1998-1999. The extract was applied as plant spray in concentrations from 0.017 to 0.099%. First treatment of rose shrubs was done when visible disease symptoms occurred on leaves and spraying was repeated 3 (in plastic tunnel) or 10-times (in the field) at weekly intervals. In the second experiment roses with visible powdery mildew symptoms were sprayed once with grapefruit extract. Leaves were sampled one or 7 days after the extract application and germination of spores of S. pannosa var. rosae on potato dextrose agar was evaluated. In the next experiment roses grown under plastic tunnel were sprayed once with the tested preparation. After 24 hours leaves were collected and appearance of fungal hyphae and spores of S. pannosa var. rosae was studied in scanning electron microscope. In the control of S. pannosa var. rosae grapefruit extract at conc. 0.066% was as effective as triforine (standard) applied at 0.027%. Reduction of concentration resulted in the decreased efficacy of the tested preparation. Spores of S. pannosa var. rosae collected one day after grapefruit extract application germinated in about 5%. Analyses of spore vitality 6 days letter showed that only about 15% of conidia could germinated on PDA agar. In contrary, spores from untreated leaves germinated in about 95%. Scanning electrone microscope analysis of leaves taken from plants protected with grapefruit extract showed that most of hyphae were separated from leaf surface. Almost all hyphae and spores were degenerated. In the control of D. rosae the preparation in all tested concentrations gave satisfactory results but was less effective than triforine.

Ascomycota↗

Fungal tunneling of hair from a buried body.

Tunnels produced in human head hair by fungal hyphae were examined with a light microscope and with a scanning electron microscope. The tunnels had small diameters and exhibited minimal branching. The use of a backscattered electron detector facilitated the locating of the openings of the tunnels in the surfaces of the hairs. In the backscattered electron image, tunnel openings appeared as dark spots. The tunneling hyphae did not show a preference for a particular location for entering the shaft of the hair. Some hyphae penetrated under the free edges of the cuticular scales, while others burrowed through the surfaces of the scales.

Female↗

[Application of methylene blue staining in detection of Malassezia].

OBJECTIVE: To assess the value of methylene blue and parker ink staining in the detection of Malassezia. METHODS: Detecting Malassezia in three dermatoses samples (pityriasis versicolor, Malassezia folliculitis and dandruff) stained by methylene blue (3 g/L) or parker ink. Comparing the effectiveness and the positive rates of Malassezia detection between the two staining methods. Staining Malassezia and Candida and comparing the specificity of the two staining methods in staining Malassezia. RESULTS: Dark blue hyphae and spores were observed in smears stained by methylene blue, and light blue hyphae and spores were observed in smears stained by parker ink. Hyphae and spores were easy to identify on the background. The positive rates of methylene blue and parker ink for detecting Malassezia in the three dermatoses were 96% and 94% respectively. There was no significant difference between the two dye liquors in the positive rate for deterting Malassezia. Both Malassezia and Candida could be stained by methylene blue; nevertheless, only Malassezia could be stained by parker ink. CONCLUSION: Both methylene blue and parker ink staining can be used to detect Malassezia, but parker ink has higher specificity in Malassezia staining.

Dermatomycoses↗

[Conidiobolomycosis: a case report with histophathologic findings].

Conidiobolomycosis is a subcutaneous mycosis of the facial midline affecting primarily adult males. It is caused by the saprophytic fungus, Conodiobolus coronatus, present in soils and dried vegetables, characteristic of intertropical regions. C. coronatus belongs to the class Zygomycetes, order Entomophthorales; it is a fungus composed of thick, short hyphae that grows at temperatures between 30 degrees C and 37 degrees C and causes nasal granulomas. The histologic findings are described of a case of conidiobolomycosis in a 31-year-old male, born and resident in the Urabá region of Colombia. He presented with a mid-facial deformity of the nose and upper lip edema, and polypoid images in the maxillary sinuses with destruction of the nasal septum. The biopsy revealed a diffuse inflammatory lesion located in the deep dermis and in the hypodermis corresponding to a necrotizing granuloma. Associated eosinophils and the presence of the Splendore-Hoeppli phenomenon were noted in the vacant central zone which apparently corresponded to location of the fungal hyphae. They did not stain with HE stain, but reacted to the PAS and Grocott staining techniques and appeared as rigid, thick-walled hyphae, centrally twisted and with cone-shaped endings.

Adult↗