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[Ecological and epidemiological relationships between Sporothrix schenckii, a fungus pathogenic for man and the Ceratocystis genus].

Sporothrix schenckii is a pathogenic fungus for man and animal. Its perfect form is not yet known. Many studies have been made to compare this fungus to ascomycetes belonging to the genus Ceratocystis. This paper summarizes some important data about the ecology and the epidemiology of the fungal complex Sporothrix-Ceratocystis. Previous results obtained by several groups of workers are strongly in favor of a relationship between the two genera. The epidemiological studies carried out in different areas have shown that Sporothrix schenckii and Ceratocystis stenoceras are frequently isolated from the same fragments of plants such as pine or eucalyptus. According to the time and areas, it appears that there is or there is not a relation between the presence of the fungus on the plants or in the soil and the disease. The former case is well illustrated by the epidemic of the Transvaal between 1940 and 1944. Sporothrix schenckii has been found to be present either in endemic areas like Guatemala (around the Ayarza Lake) or in Alsace, or in Corse where no case has been reported for a long time. In France, except for the curious epidemic observed between the years 1903 and 1912, no or very rare cases have been observed although Sporothrix schenckii is abundant in the soil and injuries with contaminated materials are probably extremely frequent. The presence or even the abundance of the fungus in nature is not enough to explain the development of the disease. Some not well known factors are necessary. So as for all disease, the immunological reaction of the host is important and the defects of these mechanisms must be suspected.

Ascomycota

The combined effect of a cutaneo-lymphatic fungus, Sporothrix schenckii and a lymphatic-dwelling nematode, Brugia Malayi.

Domestic cats were infected with third-stage Brugia malayi in such a way that the parasites were restricted to the regional lymphatics of one hind limb. Later, these cats were exposed on the same leg to the yeast phase of Sporothrix schenckii. Edema and fibrosis were more extensive in cats infected with both Brugia and Sporothrix than in cats with either of these organisms alone. Lesions tended to appear earlier, more consistently and progressed more rapidly in cats with dual infections than in Sporothrix controls. No septic lesions were seen in any Brugia controls. Dissemination of the organism was also greater in animals with dual infections than in those infected only with Sporothrix. On the basis of this study, it appears probable that in certain istances fungi, as well as bacteria, may play an important role in transforming a mild subclinical case of lymphatic filariasis into a serious medical problem and that a filarial infection may greatly exacerbate an existing Sporothrix infection.

Animals

[Occurrence of neuraminidase in Sporothrix schenckii and Ceratocystis stenoceras and its role in ecology and pathomechanism of these fungi (author's transl)].

Some strains of Sporothrix schenckii and one strain of Ceratocystis stenoceras were investigated and the occurrence of the enzyme neuraminidase (= sialidase, N-acetylneuraminate glycohydrolase, EC 3.2.1.18) was proved not only by Warren's thiobarbituric acid assay but also by immunoelectrophoresis technique using monospecific antisera against 25 human glycoproteins, lipoproteins, immunoglobulins, fibrinogen and some other proteins (Table 1). There are also fibrinolytic and lipolytic enzyme activities in strains of Sporothrix chenckii corresponding with similar findings of other authors and also in the Ceratocystis stenoceras strain. The temperature optimum of production of neuraminidase seems to be lower than 37 degrees C (Table 2) and all strains failed to produce the enzyme after cultivation at 37 degrees C during some months. The findings of the existence of neuraminidase in the both species confirm also their close relationship. Because the occurrence of N-acylneuraminate was shown only in animals and in some microorganisms but not in plants it can be suggested that also Sporothrix schenckii and Ceratocystis stenoceras grow on animal or neuraminate containing microbial material but not on plants as it is assumed hitherto. Corresponding to this hypothesis the pathogenicity of Sporothrix schenckii and also the facultative pathogenicity of Ceratocystis stenoceras are to see in connexion with the action of neuraminidase and other enzymes metabolizing animal substrates. It can be concluded that neuraminidase is a possible additional pathogenic factor in sporotrichosis below 37 degrees C.

Ascomycota

Ex vivo determination of potentially virulent Sporothrix schenckii,.

Hyphae from 30 isolants of Sporothrix and Ophiostoma species were washed, dried and pyrolyzed at 350 degrees C. Pyrolysis products were separated on a Carbowax column heated 7.5 degrees C/min to and maintained for 50 min at 160 degrees C. Hydrogen flame detector responses were recorded graphically. Fifteen clinical isolants of S. schenckii from geographically separated sources produced qualitatively identical pyrograms. S. foliorum, 8 avirulent S. schenckii and other Sporothrix species isolants from soils, and Sporothrix states of 6 Ophiostoma species yielded pyrograms readily distinguished from each other and from those of virulent S. schenckii. Taxonomic and clinical implications of the pyrograms are mentioned.

Ascomycota

Granulomatous synovitis and osteitis caused by Sporothrix schenckii.

Sporotrichosis must be considered in the differential diagnosis of granulomatous inflammation involving bones and joints. The organisms are difficult to demonstrate in direct smears and in histiologic sections, but they grow readily on routine fungal culture media. The cases of two patients, one with sporothrix arthritis and one with sporothrix arthritis and osteitis, are presented. The latter patient underwent ten surgical procedures over a period of 6 1/2 years and was treated for tuberculous arthritis without a definite diagnosis before fungal cultures were obtained and Sporothrix schenckii isolated.

Diagnostic Errors

Isolation and characterization of Sporothrix schenckii from clinical and environmental sources associated with the largest U.S. epidemic of sporotrichosis.

The largest recorded epidemic of sporotrichosis in the United States occurred in 1988 and involved a total of 84 cases in 15 states. All cases were associated with Wisconsin-grown sphagnum moss. Twenty-one clinical isolates of Sporothrix schenckii and 69 environmental isolates of Sporothrix spp. from the epidemic were characterized and compared. The environmental isolates were recovered from 102 samples of sphagnum moss and other material by using direct plating techniques. Characteristics examined included macroscopic and microscopic morphology, conversion to a yeast phase, exoantigen reactions, and virulence in mice. On the basis of these studies, eight environmental isolates were identified as S. schenckii, five were identified as Ophiostoma stenoceras, and the remainder were identified as Sporothrix species. The environmental isolates of S. schenckii were recovered from moss samples from one Pennsylvania nursery and from three New York State Soil and Water Conservation districts, but none were recovered from moss directly from the bogs in Wisconsin.

Disease Outbreaks

Sporothrix tenosynovitis--differential diagnosis of granulomatous inflammatory disease of the joints.

Sporothrix tenosynovitis is difficult to distinguish from rheumatoid arthritis--the organisms are scarce in biopsy specimens, and both diseases cause a granulomatous synovitis which can affect multiple joints, have associated subcutaneous nodules, similar radiographic findings, and respond to heat and salicylates. This report describes a patient treated for over 2 years as having rheumatoid arthritis. Multiple tenosynovectomy specimens revealed granulomatous synovitis with sterile cultures. Eventually a fungal culture was positive for Sporothrix. This report discusses the histopathologic features of Sporothrix synovitis, and stresses the importance of repeated fungal cultures from patients with granulomatous synovitis unresponsive to steroids.

Antifungal Agents

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

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

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

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

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

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

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

Effects of zinc on macromolecular synthesis and nuclear division during the yeast to mycelium transition in Sporothrix schenckii.

Zinc ions (10 mM) have been reported previously to inhibit the yeast to mycelium transition in Sporothrix schenckii. Yeast cells of this fungus were harvested, selected by filtration and allowed to form germ tubes in a basal medium with glucose in the presence of 10 mM zinc and the effects of this ion on protein, RNA and DNA synthesis and nuclear division recorded. All of these processes were affected by the addition of 10 mM zinc to the medium. Nevertheless, the inhibition of protein synthesis was observed earlier than that of RNA or DNA synthesis and was of a greater magnitude than that observed for both of these processes. Protein synthesis was inhibited within the first hour after inoculation, at which time this process begins in the control cells. RNA synthesis was inhibited during the 3 to 6 h interval after inoculation, that is, 3 h after the start of this process in the control cells. After 9 h of incubation, the inhibition of protein synthesis had reached its maximum at 70%, while that of RNA synthesis was only 52%. DNA synthesis was slightly inhibited, with maximum inhibition being observed 9 h after inoculation. Nuclear division in cells forming germ tubes in the presence of 10 mM zinc took place with a 3 h delay in relation to the control cells. These observations suggest that the inhibition of protein synthesis might be the most important mechanism by which zinc inhibits the yeast to mycelium transition in S. schenckii.

Cell Division

Detection of cellular immunity with the soluble antigen of the fungus Sporothrix schenckii in the systemic form of the disease.

Sporothrix schenckii is the etiologic agent of sporotrichosis, a mycosis of world-wide distribution more commonly occurring in tropical regions. The immunological mechanisms involved in the prevention and control of sporotrichosis are not fully understood but apparently include both the humoral and cellular responses. In the present investigation, cellular immunity was evaluated by in vivo and in vitro tests in mice infected with yeast-like forms of S. schenckii. The disease developed systemically and cellular immunity was evaluated for a period of 10 weeks. The soluble antigen utilized in the tests was prepared from yeast form of the fungus through the sonication (20 min: 10 sonications at 50 W at 2-min intervals). Delayed hypersensitivity and lymphocyte transformation tests showed that the cellular immune response was depressed between the 4th and 6th week of infection when the animals were challenged with the soluble fungal antigen. This depression frequently indicates worsening of the disease, with greater involvement of the host. This is a promising field of research for a better understanding of the pathogeny of this mycosis.

Animals

Experimentally induced mycetoma: species of Sporotrichum and Sporothrix.

The potential pathogenicity of four saprophytic species of Sporotrichum, viz; S. carnis, S. cerebriforme, S. pruinosum and S. thermophile were compared to S. schenckii and all could survive in vivo and S. pruinosum may possess limited virulence. Thielavia thermophila and Sporothrix albicans were able to cause local mycetoma.

Animals

Spore ultrastructure in Sporothrix schenckii.

Pathogenic strains of Sporothrix schenkii may show triangular spores, whose angular shape is maintained by a tie-beam effect in the inner cell wall structure. This difference in wall structure lies adjacent to a folded and possibly more active part of the spore cytoplasm. The supposed generation of asci in old cultures was simulated by the death of hyphae which are reinvaded by intrahyphal growth with intrahyphal spore production, while true asci were not seen.

Cell Membrane