[Results of cultural vaginal smear studies as a proof of pathogenous Blastomyces. Laboratory diagnosis of Blastomyces infection of the female genitalia].
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Bacteriological Techniques↗
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[Comparative studies on the occurrence of blastomyces on healthy and sick skin. 2. Epidemiology of propagation of blastomyces on healthy and sick skin].
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Blastomyces↗
[Comparative studies on the occurrence of blastomyces on healthy and diseased skin. 1. The occurrence of blastomyces on healthy and diseased skin].
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Blastomyces↗
[Infusion liquids as the cause of blastomyces septicemia in childhood. On the development, management and prevention of iatrogenic blastomyces septicemia].
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Amphotericin B↗
Characterization of uptake of 2-deoxy-2-[18F]fluoro-D-glucose by fungal-associated inflammation: the differential uptake ratio for blastomyces-associated lesions is as high as for lymphoma and higher than for turpentine abscesses in experimentally induced lesions in rats.
PURPOSE: The objective of this investigation was to determine the biologic basis and the significance of uptake of 2-deoxy-2-[18F]fluoro-D-glucose (FDG) using experimentally created fungal lesions in rats. PROCEDURES: Uptake of FDG by experimentally induced Blastomyces granulomas was compared with uptake by turpentine abscesses (Group 1) and by lymphomas (Group 2) using the differential uptake ratio (DUR) measured one hour after administration of 2 mCi FDG intravenously. Frozen tissue sections of Blastomyces lesions and turpentine abscesses were placed in contact with radiographic film for macroautoradiography. RESULTS: In rats in Group 1, the median (range) DUR for the Blastomyces granulomas was 1.9 (1.1-2.6) and was significantly higher than the DUR for turpentine abscesses 0.9 (0.6-1.4) and muscle 0.2 (0.1-0.5; P < 0.001). In Group 2, the median (range) DUR for the Blastomyces granulomas, lymphomas, and muscle from the rats in Group 2 were 1.8 (1.2-3.4), 1.9 (1.0-4.0), and 0.2 (0.1-0.3), respectively. There was no significant difference between the DUR of Blastomyces granulomas and lymphomas. Macroautoradiographs of the Blastomyces granulomas revealed intense uptake of FDG in the region occupied by the yeast organisms and the granulomatous inflammation. CONCLUSIONS: Blastomyces granulomas typically have high uptake of FDG associated with the region composed of the granulomatous inflammatory reaction and Blastomyces yeast organisms.
Journal Article↗
A technique for creating localized subcutaneous Blastomyces granulomas in rats.
Our purpose was to develop a simple, reliable method for creating subcutaneous Blastomyces dermatitidis nodules in rats and to describe the histologic appearance of these lesions. We used B. dermatitidis isolated from a dog with blastomycosis to prepare a Blastomyces yeast suspension. Four rats were used to test initial dose concentrations of 10(5), 10(6), 10(9), and 10(10) yeast organisms. The dose was administered subcutaneously over the distal tibia in a volume of 0.1 ml. We then inoculated 35 additional rats with 10(9) or 10(10) yeast organisms. Rats were euthanized 7, 10, 14, 21, or 28 days after inoculation, and the histologic appearance of the nodules was described. A full post-mortem examination sought evidence of systemic spread of Blastomyces organisms. We successfully induced subcutaneous Blastomyces abscesses in 34 of 37 rats injected with 10(9) or 10(10) organisms. Nodules first appeared 3 to 7 days after injection and reached 2 to 15 mm in diameter by 7 to 28 days after inoculation. Histologically the lesions were characterized by a necrotic center surrounded by a layer of viable yeast and granulomatous inflammation. Live yeast organisms were recovered from all lesions. No adverse effects or systemic spread of Blastomyces organisms were observed. We conclude that subcutaneous Blastomyces abscesses can be induced safely and reliably in rats after injection of 10(9) and 10(10) organisms. Histologically, the experimentally induced lesions share both similarities to and differences with lesions of naturally occurring blastomycosis.
Animals↗
Purification and characterization of the major antigen WI-1 from Blastomyces dermatitidis yeasts and immunological comparison with A antigen.
The lack of well-defined antigens from Blastomyces dermatitidis has hampered the ability to reliably diagnose human infection and study the immunobiology of blastomycosis. We recently discovered a novel surface protein on B. dermatitidis yeasts, designated WI-1, and demonstrated it to be a key antigenic target of humoral and cellular responses during infection. In the present article, we purified and characterized WI-1 and compared it immunologically with the only Blastomyces antigen commercially available, A antigen. WI-1 was purified by high-performance liquid chromatography over a DEAE-cellulose column. It eluted from the column at a point on the salt gradient corresponding to 460 to 490 mM NaCl, reflecting its acidic pI of approximately equal to 5.2. Purified WI-1 had a molecular mass of 120 kDa and contained a large amount of cysteine (85 residues) and aromatic amino acids but undetectable carbohydrate. In contrast, A antigen had a molecular mass of 135 kDa and contained 37% carbohydrate. Immunological comparison of the two antigens showed that, when radiolabeled, WI-1 was more reactive with anti-Blastomyces antisera than A antigen but did not cross-react with anti-Histoplasma antisera. Proteinase digestion of WI-1 eliminated its recognition by anti-WI-1 and anti-Blastomyces antisera. Proteinase treatment of A antigen had no effect on its recognition by anti-Blastomyces or anti-Histoplasma antisera, but periodate treatment abolished recognition by anti-Histoplasma antisera, indicating that the cross-reactive determinant(s) of A antigen is displayed on the accompanying carbohydrate. In further studies, anti-WI-1 antiserum reacted with A antigen and, conversely, anti-A antiserum and monoclonal antibodies (MAbs) reacted with WI-1, indicating a shared determinant on the two antigens. A recombinant 25-amino-acid repeat, recently cloned from WI-1 and found to be the major target of antibody recognition of WI-1, reacted strongly with anti-A antiserum and MAbs. In MAb competition tests, MAbs specific for the 25-residue repeat abolished binding of anti-A antiserum to A antigen. In antigen inhibition tests, the recombinant repeat abolished binding of anti-A antiserum to A antigen. These results demonstrate that the repeat is the major site of antibody recognition of both WI-1 and A antigen and that the recombinant, nonglycosylated peptide could replace either native antigen in formatting better diagnostic tests for blastomycosis. Moreover, they suggest that producing fungal protein antigens as nonglycosylated peptides in a procaryotic expression system may circumvent problems of antigen cross-reactivity that are due to posttranslational modification.
Animals↗
Attempted isolation of Blastomyces dermatitidis from native shrews in northern Wisconsin, USA.
The precise ecological niche of Blastomyces dermatitidis is unknown. The related dimorphic fungus, Paracoccidioides brasiliensis, has been isolated from South American ground-dwelling insectivorous armadillos. We attempted to isolate Blastomyces from shrews, North American ground-dwelling insectivores that have been shown to harbor Histoplasma capsulatum in endemic areas. Forty-seven masked shrews (Sorex cinereus) and 13 northern short-tailed shrews (Blarina brevicauda) were collected in endemic areas of northern Wisconsin and Michigan using pitfall traps. Specimens were collected between 1998 and summer 2002, stored frozen, then necropsied. Cultures of nasopharynx, lungs, liver, spleen and large and small bowel were placed on yeast extract phosphate agar with one or two drops of ammonium hydroxide. Cultures for Blastomyces were negative from all 60 shrews and two deer mice (Peromyscus maniculatus) and three southern red-backed voles (Clethrionomys gapperi), which were trapped inadvertently. Histological examination of 36 of these specimens revealed no Blastomyces yeast forms. Northern Wisconsin shrews do not appear to be carriers of B. dermatitidis.
Animals↗
Molecular taxonomy and epidemiology of Blastomyces and Histoplasma species.
Cladistic analysis of partial 26S rRNA sequences was used to estimate evolutionary distances among species and varieties of the dimorphic onygenalean genera Blastomyces, Coccidioides, Emmonsia, Histoplasma and Paracoccidioides. With the exception of Coccidioides, all genera were closely related, with about 5% base differences and even less (1-2%) between Blastomyces and Emmonsia. These data were supported by a teleomorph in the same genus Ajellomyces. In a phylogenic study of a wide range of ascomycete orders and families, Coccidioides immitis was found to be closest to Aphanoascus fulvescens and Chrysosporium keratinophilum, and to have relative distances to the remaining dimorphic genera (family Onygenaceae) similar to those of the dermatophytes (family Arthrodermataceae). The sequencing data were confirmed by genomic comparisons. All dimorphic genera had a nuclear DNA base composition in the same range of 46.6-47.3% G + C. The DNA melting curves of Blastomyces and Histoplasma strains showed irregularities that were ascribed to the presence of AT-rich stretches in satellite DNA rather than in mitochondrial DNA. Derivative profiles proved to be highly reproducible within regional populations and coincided with differences in clinical behaviour of each species. Blastomyces dermatitidis generated two kinds of curves, corresponding to the geographically distinct serotypes 1 and 2. The African type (serotype 2) was characterized by a classical sigmoidal melting curve similar to that for all strains of Coccidioides, Emmonsia and Paracoccidioides. In contrast, the American type (serotype 1) contained satellite DNA (27% G + C). A rRNA base difference of 1.5% was observed between geographical types, a value slightly higher than that noted between Histoplasma capsulatum and its variety farciminosum (0.9%). All three H. capsulatum varieties presented irregularities in their DNA melting curves. The molecular data support the recognition of two of them as agents of blastomycosis and the assignment of more than one species and two varieties to the genus Emmonsia.
Animals↗
In vivo and in vitro cell-mediated immune responses to a cell wall antigen of Blastomyces dermatitidis.
An alkali-soluble, water-soluble cell wall fraction of Blastomyces dermatitidis, designated B-ASWS, was evaluated as an antigen for detecting in vivo (skin tests) and in vitro migration inhibition factor (MIF) production and lymphocyte transformation (LT) responses in Blastomyces-infected guinea pigs. The biological activity of B-ASWS was compared with that of blastomycin KCB-26. The superiority of B-ASWS, in terms of its sensitivity and specificity, was evident in in vivo and in vitro assays. Skin tests responses were obtained in 21 of the 24 Blastomyces-infected guinea pigs, whereas only one of the 14 Histoplasma-infected guinea pigs were significantly greater than those obtained using cell populations from Histoplasma-infected or noninfected guinea pigs. The con-MIF and LT in peritoneal exudate cells and lymph node cells of homologuosly infected animals. In each biological system, the response of the Blastomyces-infected guinea pigs were significantly greater than those obtained using cell populations from Histoplasma-infected or non-infected guinea pigs. The contrasting efficacy of B-ASWS as compared with blastomycin KCB-26, suggests that the cell wall antigen will be a useful tool for detecting cell-mediated immune responses in blastomycosis.
Animals↗
Cutaneous lesions showing giant yeast forms of Blastomyces dermatitidis.
BACKGROUND: The yeast forms of Blastomyces dermatitidis usually range from 8 to 15-20 micro m in diameter. Larger yeast forms have previously been reported only twice in immunosuppressed patients. In both patients these large forms were seen within the lung. CASE REPORT: We present a 14-year-old cardiac transplant patient, who presented 36 days following his transplantation with acute respiratory distress followed a few days later by erythematous cutaneous papules. RESULTS: Biopsy of a skin lesion showed yeast forms, some greater than 40 micro m in diameter, within and surrounding dermal vessels. Cultures later grew Blastomyces dermatitidis. CONCLUSIONS: To our knowledge this is the first reported case of giant forms of Blastomyces dermatitidis within the skin. With increased iatrogenic immunosuppression, we may expect to see more diverse morphologic forms with deep fungal infections.
Adolescent↗
Intracellular growth and phagocytosis of Blastomyces dermatitidis by monocyte-derived macrophages from previously infected and normal subjects.
Blastomyces dermatitidis evokes responses of human cellular immunity typical of other intracellular fungal pathogens. Differences in growth rates of intracellular Blastomyces yeast and the differences in amounts of yeast phagocytized by macrophages were determined for macrophages derived from peripheral blood monocytes from 11 persons with treated blastomycosis and 11 normal, healthy persons. Cellular immunity was examined by lymphocyte uptake of [3H]thymidine in response to a specific antigen of Blastomyces yeast. Yeast were more readily phagocytized by macrophages from the previously treated donors when compared with those from the normal donors; the yeast were confirmed to be intracellular by transmission electron microscopy. Likewise, a decrease in growth rates of yeast was demonstrable in cultures of macrophages from previously treated donors as compared with normal donors. This greater efficiency of phagocytosis and growth inhibition of B. dermatitidis reflects another mechanism of human cellular immunity to this fungal infection.
Blastomyces↗
Passive transfer of delayed hypersensitivity to Blastomyces dermatitidis between mice.
This study further characterized the delayed hypersensitivity state induced in animals by Blastomyces dermatitidis exposure. Passive transfer of delayed hypersensitivity by transfer of cells and inhibition of migration of peritoneal exudate cells were studied, using sensitized mice of two inbred strains. Donor mice were subcutaneously inoculated with viable B. dermatitidis yeast cells. After 15 days, spleen cells or serum from these animals were injected intravenously into normal recipients of the same strain. After 24 h these mice were footpad tested with killed B. dermatitidis yeast cell antigen. Mice receiving spleen cells from sensitized animals had a significant increase in footpad thickness 24 to 48 h after testing. Those receiving only serum remained negative. Migration of peritoneal exudate cells from blastomyces-sensitive donor mice was inhibited by presence of blastomycin but not by mycobacterial antigen. Neither blastomyces-sensitive nor control animals reacted to footpad or migration inhibition testing with mycobacterial antigen.
Animals↗
Growth inhibition of Blastomyces dermatitidis in alveolar and peripheral macrophages from patients with blastomycosis.
Blastomyces dermatitidis evokes a pyogranulomatous disorder with organisms frequently found inside giant cells. Macrophages from bronchoalveolar lavage fluid and peripheral blood in monolayer cell cultures were challenged with live yeast organisms to examine phagocytosis and intracellular growth. A greater number of macrophages from patients recovering from blastomycosis had phagocytized Blastomyces compared with macrophages from healthy control donors. No differences were detected within the groups between alveolar and peripheral macrophages. Intracellular growth of the fungus was reduced in cultures of both cell types from patients compared with those from control subjects. Supernatants from specific Blastomyces antigen-stimulated lymphocyte cultures were collected, and treatment with the supernatant to control donors' macrophages resulted in increased phagocytosis and inhibition of intracellular growth. Antigen-induced lymphocyte stimulation as a correlate of cellular immunity is qualitatively related to alveolar or peripheral macrophage phagocytosis and growth inhibition of this fungus.
Blastomyces↗
Identification of Blastomyces dermatitidis in the stool of a dog with acute pulmonary blastomycosis.
We report the identification of Blastomyces dermatitidis by microscopic examination of a direct faecal smear from a dog with pulmonary blastomycosis. A simultaneously obtained faecal culture grew Blastomyces dermatitidis. The fungus was also cultured from a transtracheal sample from this same dog. This report suggests that yeast-phase cells of B. dermatitidis may be recovered in the stool of dogs with pulmonary blastomycosis following transit through the gastrointestinal tract of swallowed infected sputum. Implications regarding the ecology of Blastomyces dermatitidis are discussed.
Animals↗
RAPID QUANTITATIVE METHOD FOR MEASURING THE SENSITIVITY OF BLASTOMYCES DERMATITIDIS TO FUNGISTATIC AGENTS.
Guidry, D. J. (Louisiana State University School of Medicine, New Orleans) and Findlay Maier. Rapid quantitative method for measuring the sensitivity of Blastomyces dermatitidis to fungistatic agents. J. Bacteriol. 85:504-508. 1963.-Isolates of Blastomyces dermatitidis (yeast phase) were grown in continuous shake liquid culture. When sufficient cells were added to impart an initial optical density of 0.100 to the inoculated medium, growth was rapid, and the time required to assay fungistatic agents was reduced to 48 hr. Standard curves on light transmittance and direct cell count facilitated preparation of inoculum and measurement of growth. Generation time was determined for various growth phases of B. dermatitidis, and the data were applied to selection of the optimal period of growth for assaying fungistatic agents. Four isolates of B. dermatitidis were tested for their sensitivity to amphotericin and 2-hydroxystilbamidine. An amphotericin B concentration of 1.4 mug/ml was sufficient for maximal inhibition of all isolates tested; there were differences among isolates in sensitivity to lower concentrations of the drug. A 2-hydroxystilbamidine concentration of 62.5 mug/ml was sufficient for maximal inhibition of two isolates; the two remaining isolates required a concentration of 125 mug/ml. A latent period in inhibition of growth occurred when amphotericin B and 2-hydroxystilbamidine were added to cultures of B. dermatitidis. This latent period, together with generation time of the yeast cells, determined the maximal degree of inhibition which could be attained.
Amphotericin B↗
LIPID CONTENT OF FOUR STRAINS OF BLASTOMYCES DERMATITIDIS OF DIFFERENT MOUSE VIRULENCE.
Di Salvo, Arthur F. (Medical College of Georgia, Augusta) and J. Fred Denton. Lipid content of four strains of Blastomyces dermatitidis of different mouse virulence. J. Bacteriol. 85:927-931. 1963.-Four strains of Blastomyces dermatitidis showing marked differences in virulence for mice were studied to determine whether there was any relationship between lipid content and virulence. Replicate samples of heat-killed yeast and mycelial cells, after rupturing by grinding, were extracted with a chloroform-methanol mixture (2:1) in a Soxhlet apparatus for 48 hr. Total extractable lipid was computed by evaporating samples of the extracts to dryness and weighing; the phospholipid fraction was determined by a modified photometric technique. The amount of lipid in the yeast phase of each strain was relatively constant and characteristic as determined by extraction of four samples of each. The extractable lipid increased from 6.8% in the avirulent strain Ga-1 to 12.3% in the highly virulent strain SL-1, the lipid content correlating directly with increased mouse virulence. There was no similar correlation of phospholipid content with mouse virulence. The mycelial phases of two strains contained more than double the lipid and phospholipid present in the corresponding yeast phases. Data presented suggest that the lipid fraction of the yeast phase of B. dermatitidis, in addition to being related to the granulomatous reaction, is possibly related to virulence.
Animals↗
SEROLOGICAL CHARACTERIZATION OF PATHOGENIC FUNGI BY MEANS OF FLUORESCENT ANTIBODIES. I. ANTIGENIC RELATIONSHIPS BETWEEN YEAST AND MYCELIAL FORMS OF HISTOPLASMA CAPSULATUM AND BLASTOMYCES DERMATITIDIS.
Kaufman, Leo (Communicable Disease Center, Atlanta, Ga.) and William Kaplan. Serological characterization of pathogenic fungi by means of fluorescent antibodies. I. Antigenic relationships between yeast and mycelial forms of Histoplasma capsulatum and Blastomyces dermatitidis. J. Bacteriol. 85:986-991. 1963.-Four antiglobulins prepared against the yeast and mycelial forms of Histoplasma capsulatum and Blastomyces dermatitidis were labeled with fluorescein isothiocyanate. Cross-staining and adsorption techniques enabled the discernment of five antigenic factors in the fungi studied. The yeast-phase cells of H. capsulatum and B. dermatitidis contained the respective distinct antigens B and D, and a common factor E. In addition, the yeast-phase cells of these two species shared antigen C with the mycelial elements of H. capsulatum. The mycelial cells of H. capsulatum and B. dermatitidis shared antigen A with the yeast-phase cells of these two organisms and contained no distinct antigens.
Antigens↗