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PubMed1979

Isolation of Sporothrix fungorum from a 500-year-old mummy found in Greenland.

A 500-year-old mummy of a seven-month-old Eskimo infant recently removed from a tomb north of Umanak presented problems of preservation. Among these was fungal coating on the skin, especially on the face. The fungus was isolated and diagnosed as a Sporothrix fungorum. The skin was successfully treated with nystatine. The species Sporothrix fungorum isolated is considered to be very rare, having been reported previously only from Europe and South Africa, but never from Greenland.

Greenland↗
PubMed1989

Immunoblot analysis of antibody responses to Sporothrix schenckii.

The serologic response to Sporothrix schenckii was investigated in patients with sporotrichosis by solid-phase enzyme-linked immunosorbent assays (ELISAs) and Western immunoblot techniques. A soluble antigen preparation derived from an S. schenckii isolate contained 15 protein staining components ranging in molecular size from 22 to 70 kilodaltons (kDa) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Sera from 40 patients with sporotrichosis demonstrated Sporothrix immunoglobulin G antibody by ELISA with titers between 128 and 65,200. No sera from 300 healthy individuals or 100 patients with various systemic mycoses other than sporotrichosis had ELISA titers greater than 64. By Western immunoblotting of the antigens separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, sera from 10 patients with cutaneous sporotrichosis reacted with 8 to 10 antigen components (range, 40 to 70 kDa), while sera from 15 patients with extracutaneous sporotrichosis reacted with a greater number of antigen components (15 to 20 bands) over a wider range of molecular sizes (22 to 70 kDa). Antibody to 40- and 70-kDa antigen components was detected by immunoblots in all sera tested from patients with sporotrichosis. Antibody to 22- to 36-kDa antigen components was present in sera from 13 of 15 patients with extracutaneous sporotrichosis, but these lower-molecular-weight components were not detected by sera from patients with cutaneous sporotrichosis. Antibody to these components was not detected by Western blotting in sera from 19 of 20 patients with other fungal diseases or from 30 healthy individuals. Purification of these specific antigen fractions could provide the basis of a sensitive and specific serodiagnostic test to indicate the presence and activity of extracutaneous sporotrichosis.

Antibodies, Fungal↗
PubMed1992

Use of a mouse model to evaluate clinical and environmental isolates of Sporothrix spp. from the largest U.S. epidemic of sporotrichosis.

Five clinical and 69 environmental isolates from the largest U.S. epidemic of sporotrichosis were evaluated in NYLAR male mice following intravenous injection of 5 x 10(6) to 2 x 10(8) conidia per mouse. The clinical isolates and eight environmental isolates produced 100% mortality in groups of three mice each between 12 and 24 days after injection. These virulent isolates grew at 37 degrees C, were dematiaceous by virtue of melanin (melanized) on permissive media (e.g., potato dextrose agar), produced ovoid conidia borne sympodially on lateral conidiophores and pleurogenously about the main hyphal axis, and were identified as Sporothrix schenckii. Two melanized environmental isolates that grew at 35 degrees C but not at 37 degrees C were not virulent and had subtle morphological differences from S. schenckii. The remaining environmental isolates were not melanized, were not virulent, and were not S. schenckii; five were identified as Ophiostoma stenoceras and the remainder were identified as Sporothrix spp. Quantitative organ cultures revealed that clinical isolates grew exponentially in livers and testes, in contrast to an isolate of O. stenoceras that was eliminated from liver, lung, and spleen but that persisted in the testes throughout the 14-day sample period. This model helped to confirm the identification of S. schenckii isolates obtained from the environment.

Animals↗
PubMed2002

Phenotyping and genotyping of Sporothrix schenckii isolates according to geographic origin and clinical form of Sporotrichosis.

Sporothrix schenckii isolates of fixed and lymphocutaneous clinical forms from Mexico (MX), Guatemala (GT), and Colombia (CO) as well as environmental isolates from MX were studied by analyzing their phenotypic characteristics (conidial length, thermotolerance by percent growth inhibition [GI] at 35 and 37 degrees C, median lethal dose [LD(50)]) and genotypic characteristics (by random amplified polymorphic DNA [RAPD] analysis-PCR). A significant difference (P < 0.01) in the mean conidial length of S. schenckii clinical isolates from CO ( = 4.03 +/- 1.04 microm) compared with those of clinical isolates from MX ( = 2.06 +/- 0.53 microm) and GT ( = 2.68 +/- 0.83 microm) was observed. The lowest thermotolerance, as determined by measurement of percent GI, was exhibited by isolates from CO at 35 degrees C ( = 50.1% +/- 15.9%) and 37 degrees C ( = 72.7% +/- 10.9%). In general, the highest virulence, as determined by measurement of the LD(50) for mice, was observed for the MX environmental isolates. RAPD analysis-PCR with 10-mer primers OPBG-01, OPBG-14, and OPBG-19 generated 52 reproducible bands. The 44 Sporothrix isolates fell into four major groups by hierarchical cluster analysis. The first group (group I), formed by 25 (of 27) isolates from MX, had two subgroups: subgroup Ia with 10 environmental isolates and subgroup Ib with 14 clinical isolates. The second group (group II) had two subgroups: subgroup IIa, formed by isolates from CO, and subgroup IIb, formed by isolates from GT. Groups III and IV each had only one clinical isolate from MX. A principal-component analysis of the same data yielded three distinct groups, depending on the geographical origins of the isolates, including the isolates in groups III and IV from MX, which were grouped with the isolates from MX by principal-component analysis. This study revealed that isolates from CO had low thermotolerances at 35 and 37 degrees C and could be associated with superficial skin lesions in patients with fixed clinical forms of sporotrichosis, the most frequent form of the disease in CO. Distinct patterns dependent on geographical origins were also revealed by RAPD analysis-PCR, but these had no relation to the clinical form of the disease.

Colombia↗
PubMed1987

Freeze-fracture electron microscopic studies of age-related plasma membrane changes in Sporothrix schenckii.

Characteristics of the plasma membrane of Sporothrix scheckii cells as revealed by freeze-fracture techniques have been classified into eight types (Y1, Y2a, Y2b, Y3a, Y3b, Y4a, Y4b, and Y5) in yeastlike cells grown under the following two conditions: brain heart infusion agar medium at 27 degrees C, and brain heart infusion agar medium at 37 degrees C. Type Y1 cells are yeastlike cells having smooth plasma membranes without any invagination. Typical characteristics of the other types are as follows: type Y2a, smooth plasma membranes with few trenchlike invaginations; type Y2b, wavy plasma membranes with few oval or irregularly formed invaginations; type Y3a, plasma membranes with many randomly distributed trenchlike invaginations; type Y3b, plasma membranes with many cocoonlike or irregularly formed invaginations; type Y4a, plasma membranes with longer trenchlike invaginations; type Y4b, plasma membranes with irregularly formed, enlarged invaginations; and type Y5, smooth or wavy plasma membranes with aggregations of intramembranous particles and with many vacuoles between cell walls and plasma membranes or in the cytoplasm in some cells. By counting the proportion of each type of yeastlike cell under the two conditions and with different cultivation periods, it appears that plasma membrane types change as aging progresses in the following order: type Y1, Y2a, Y3a, Y4a, and Y5 in conidia and type Y1, Y2b, Y3b, Y4b, and Y5 in yeastlike vegetative cells. These observations provide us with an important advantage when studying the effects of antifungal agents on the plasma membrane of Sporothrix scheckii, as it is important to know the natural course of changes in membrane structure during aging.

Cell Membrane↗
PubMed2000

[Molecular epidemiology of Sporothrix schenckii].

Restriction fragment length polymorphism (RFLP) in mitochondrial DNA (mtDNA) of clinical and environmental isolates of Sporothrix schenckii was investigated. Among mtDNA RFLP patterns with Hae III, of 14 environmental isolates morphologically identified as S. schenckii, only 2 isolates were confirmed as S. schenckii, while of more than 500 clinical isolates, all were confirmed to have this condition. Therefore, RFLP analysis of mtDNA is essential for the identification of environmental, but not clinical, isolates of S. schenckii. Isolates of Sporothrix schenckii were classified into 23 mtDNA types (Types l-23) based on mtDNA RFLP patterns with HaeIII and clustered into two major groups by phylogeny, Group A (Types 1-3, 11, 14-19, 22 and 23) and Group B (Types 4-10, 12, 13, 20 and 21). Group A isolates are predominant in South Africa, North America, Central America and South America, while Group B isolates are predominant in Australia and Japan. In Japan, the relative distribution of the mtDNA types varied with geographic region: Types 4, 6 are comparatively abundant in West Japan (Kansai and Kyushu districts), Type 5 is comparatively abundant in East Japan (Tokai, Kanto and Tohoku districts) and Type 2 is abundant in the Hokuriku district. Type 1 is found only in the Hokuriku district.

Africa↗
PubMedDate not supplied

[Isolation and characterization of wild Sporothrix schenkii strains and investigation of sporototrichin reactors].

We conducted a study in the southern mountains of the Mexican State of Oaxaca that consisted of isolation of wild Sporothrix schenckii strains obtained from soil samples and investigation of positive reactors to skin test reaction with sprotrichin antigen. The study was conducted by means of recollection of soil samples and processing of these with dilution methods and fungal isolation in ordinary culture media Sabouraud simple Agar with and without antibiotics (SS, SA). Suspected strains underwent dimorphism, melanin formation, and virulence confirmation tests. Investigation of positive reactors to sporotrichin Y (yeast) was also conducted. Three supposed strains were identified due to their reproductive characteristics, melanin production, and virulence. In the community, 144 individuals were studied, of whom 6.25% were positive to sporotichin. Isolation of virulent strains of Sporothrix schenkii from nature (soil) and primoinfection of a percentage of the studied population were confirmed.

Adult↗
PubMed1976

Atigenic similarity between Ceratocystis species and Sporothrix schenckii as observed by immunofluorescence.

Antigenic properties of Sporothrix schenckiii and 2 species of Ceratocystis (C. stenoceras and C. ulmi) were compared by indirect immunofluorescent staining technique. Both species of Ceratocystis and 2 strains of S. schenckii reacted equally well with the unabsorbed antisera against 2 strains of S. schenckii, indicating that these three species share common antigen. The absorption experiments showed that only S. schencki human strain demonstrated a unique antigen. These results support the hypothesis of Ceratocystis-Sporothrix complex in the point that both species have one major common antigen.

Antigens, Fungal↗
PubMedDate not supplied

Possible pathogenic role for Sporothrix cyanescens isolated from a lung lesion in a heart transplant patient.

Sporothrix cyanescens is a fungus rarely isolated from human specimens. Its pathogenic role has never been demonstrated but has been postulated on the basis of its occurrence in normally sterile body sites, its isolation from debilitated individuals and its thermotolerance. A first case of nodular pulmonary lesions developing in an immunosuppressed, heart transplant patient is reported. Sporothrix cyanescens was isolated from a fine needle lung biopsy. The patient failed to respond to itraconazole therapy, whereas he was successfully treated with amphotericin B.

Amphotericin B↗
PubMed1999

Ploidy study in sporothrix schenkii

Sporothrix schenckii is a dimorphic fungus that is pathogenic for humans. No sexual cycle has been reported for this fungus and little is known of its genetic constitution. To inquire into the ploidy state of Sporothrix schenckii, different approaches were followed: DNA content during transition from conidia to yeast, survival to ultraviolet irradiation, chemical mutagenesis, and induction of mitotic recombination. No change in ploidy was detected between the conidia and yeast phases of the fungi. Resistance to cell inactivation by UV irradiation was higher in S. schenckii and in Sacharomyces cerevisiae in its diploid state than in isogenic haploids a and alpha from S. cerevisiae that were inactivated at lower doses. Two mutant phenotypes, auxotrophy and albinism, were screened after chemical mutagenesis. One-step mutagenesis with either nitrous acid or ultraviolet light was unsuccessful in inducing auxotrophy but was sufficient to induce albino colonies. Two-step mutagenesis with nitrous acid in combination with UV light was necessary to attain two auxotrophic requirements: adenine and methionine. Prototrophic and pigmented revertants behaved as heterozygotes; after exposure to UV light they gave rise to derivatives which resemble the original mutation. The experiments presented in this work suggest that S. schenckii is a diploid, although aneuploidy cannot be excluded. Copyright 1999 Academic Press.

Journal Article↗
PubMed2004

Leptographium bistatum sp. nov., a new species with a Sporothrix synanamorph from Pinus radiata in Korea.

Radiata pine (Pinus radiata) lumber from Australia, Chile, and New Zealand is imported into Korea where it represents one of the most important sources of timber. Blue stain of this timber is a serious problem for which an integrated control strategy is being developed. One of the fungi that has been isolated from stained radiata pine sapwood in Korea is a Leptographium sp. that has a distinct Sporothrix synanamorph. The aim of this study was to classify this fungus. Morphological comparisons showed that this fungus is distinct from all other species of Leptographium and especially L. elegans and L. francke-grosmanniae that also have Sporothrix synanamorphs. Comparisons of sequence data for the ITS2 and part of the 28S rDNA genes as well as the beta-tubulin gene also confirmed that this fungus represents an undescribed taxon, described here as Leptographium bistatum sp. nov.

Ascomycota↗
PubMed1990

Sporothrix schenckii--a freeze-fracture study.

Freeze-fracture electron microscopy of a pathogenic dimorphic fungus Sporothrix schenckii revealed planar views of cell structures corresponding to those described already on thin sections. In addition to the characteristic differences in cell wall thickness between conidia, yeast forms and filaments, variations in plasma membrane invaginations were found. In conidia the invaginations were short and abundant, while in yeast forms they were scarce and longer. The plasma membrane of the filaments was smooth without invaginations. No differences were found in the frequency of intramembrane particles among the three forms. In the region of the septal pore the particles were circularly arranged with a characteristic partitioning on the P and E fracture faces.

Cell Membrane↗
PubMed2001

Structure elucidation of sphingolipids from the mycopathogen Sporothrix schenckii: identification of novel glycosylinositol phosphorylceramides with core manalpha1-->6Ins linkage.

Acidic glycosphingolipid components were extracted from the mycelium form of the thermally dimorphic mycopathogen Sporothrix schenckii. Two fractions from the mycelium form (Ss-M1 and Ss-M2), having the highest Rf values on HPTLC analysis, were isolated and their structures elucidated by 1- and 2-D 13C- and 1H-nuclear magnetic resonance spectroscopy, and electrospray ionization mass spectrometry with lithium adduction of molecular ions. The structures of Ss-M1 and Ss-M2 were determined to be Manalpha1-->Ins1-P-1Cer and Manalpha1--> 3Manalpha1-->Ins1-P-1Cer, respectively (where Ins = myo-inositol, P = phosphodiester). The Manalpha1-->6Ins motif is found normally in diacylglycerol-based glycophosphatidylinositols of Mycobacteria, but this is the first unambiguous identification of the same linkage making up the core structure of fungal glycosylinositol phosphorylceramides (GIPCs). These results are discussed in relation to the structures of GIPCs of other mycopathogens, including Histoplasma capsulatum and Paracoccidioides brasiliensis.

Agaricus↗
PubMed1989

Sporothrix schenckii var luriei as the cause of sporotrichosis in Italy.

The second known case of sporotrichosis caused by Sporothrix schenckii var. luriei in a patient living in Piacenza, Italy is described. In the absence of cultures, the diagnosis was based on histologic studies. Stained tissue sections (Hematoxylin and eosin, & Gomori methenamine silver) revealed hyaline, large, thick walled tissue form cells that had divided by septation or a budding process. These forms, along with the striking "eyeglass" configuration of incompletely separated cells that were also present, are the diagnostic features of this apparently rare variety. The use of a fluorescent antibody reagent, specific for S. schenckii, confirmed the identity of the etiologic agent.

Aged↗
PubMed1991

Phylogeny and molecular epidemiology of Sporothrix schenckii in Japan.

Mitochondrial DNA(mtDNA) diversity was investigated in 257 clinical isolants of Sporothrix schenckii obtained from 4 districts in Japan. S. schenckii was classified into 10 types based on Hae III restriction profiles. Phylogeny of types constructed by the method of Fitch and Margoliash [1] on the estimated sequence divergence within mtDNA using the methods of Nei and Li [2], showed that S. schenckii are grouped into 2 clusters, one group consisting of types 1, 2 and 3, and the other group consisting of the other seven types. In addition, types 1, 2, and 5 were correlated with their geographic origin, whereas type 4 was present throughout Japan.

DNA, Fungal↗
PubMed1986

Foodborne Sporothrix schenckii: infectivity for mice by intraperitoneal and intragastric inoculation with conidia.

Experimental infections with a foodborne isolate of the fungus Sporothrix schenckii were administered to mice by intraperitoneal or intragastric injection and gavage. All injected mice showed evidence of systemic sporotrichosis. Granulomas were observed from day 3 to day 12 in the organs of neonates inoculated by injection; in mice infected by gavage, granulomas were observed only in those inoculated with 10(7) conidia. Susceptibility (based on cultural recovery) of the neonates to infections with 6 X 10(6) conidia of the fungus was 100% with intragastric injection, 91% with intraperitoneal injection, and 21 and 24% (2 X 10(7) conidia) with oral intubation. With both intragastric (59%) and intraperitoneal (25%) injections, more neonates died or were cannibalized by the mother than with intubation (14.5%). S. schenckii infected neonatal mice and caused illness by the oral route as well as by injection into the tissues or stomach. Adult mice, however, were susceptible to S. schenckii only by injection into the tissues, but not by gavage.

Animals↗
PubMed1988

Analysis of restriction profiles of mitochondrial DNA from Sporothrix schenckii and related fungi.

Restriction profiles by HaeIII of mitochondrial DNA were studied for classification and distinction of Sporothrix schenckii (100 strains), S. schenckii var. luriei (1), S. curviconia (1), S. inflata (7), Ceratocystis stenoceras (17) and C. minor (7). These 6 species showed unique restriction profiles which could be discriminated from each other S. schenckii was further separable into 11 types, S. inflata into 4 types, C. stenoceras into 4 types and C. minor into 7 types based on restriction profile heterogeneity.

Ascomycota↗
PubMed1987

Ultrastructure of Sporothrix schenckii treated with iodine-potassium iodide solution.

The ultrastructural changes produced by iodine-potassium iodide solution on yeast cells of Sporothrix schenckii were investigated by transmission electron microscopy in order to clarify the mechanism of oral potassium iodide therapy for sporotrichosis. Yeast cells were dipped with solutions containing various concentrations of iodine. The rate of germination decreased markedly between the range of iodine concentrations from 0.63 microgram/ml to 5.0 micrograms/ml. No significant ultrastructural changes were seen at the concentration of the iodine of 1.25 microgram/ml (80% germination) or less. In the concentration of 2.5 micrograms/ml (50% germination), normal cells and degenerated cells coexisted. When the cells were treated with 5.0 micrograms of iodine per ml (0% germination) or more, their interior structures were completely destroyed. It is assumed that iodine treatment of the organism causes rapid destruction in the whole cell.

Iodine↗