Deoxyribonucleic acid hybridization studies of Exophiala dermatitidis and Exophiala jeanselmei

Exophiala dermatitidis and Exophiala jeanselmei share similar morphological features and have been confused with each other. To clarify the relationship between the two fungi, we conducted a deoxyribonucleic acid (DNA)-DNA hybridization study using a dot blot method. Between E. dermatitidis and E. jeanselmei, only a very low level of DNA relatedness was seen and it was confirmed that these two fungi are distinct species based on DNA similarity. Close correspondence of DNA from the isolates of E. dermatitidis was obtained, whereas the isolates of E. jeanselmei were divided into 6 groups according to their DNA similarity and a possibility was shown that E. jeanselmei is composed of genetically heterogeneous groups. The subdivision of the species E. jeanselmei by the DNA-DNA hybridization method was in agreement with serotyping exoantigens. This result suggests that DNA-DNA hybridization studies provide an excellent tool for the identification and grouping of pathogenic dematiaceous fungi.     

An overview of macrophage-fungal interactions

A review of the literature (148 references) on the interactions of fungi with polymorphonuclear cells, monocytes and macrophages is presented. The interactions of Aspergillus species, Coccidioides immitis, Blastomyces dermatitidis, Histoplasma capsulatum, Cryptococcus neoformans, Candida albicans, and Candida species with human and experimental animal derived immune cells are examined in this overview. An effort has been made to present the reader with a comprehensive list of references with the intent of encouraging additional reading and research in this important area.     

Molecular basis of pathogenicity in Blastomyces dermatitidis: the importance of adhesion

An understanding of the molecular bases of pathogenicity in Blastomyces dermatitidis and related systemic dimorphic fungi has been limited until recent years. Yeast cells of B. dermatitidis display an adhesion promoting protein termed WI-1. Recent studies entailing homologous gene targeting and mutation of WI-1 have provided null mutants at this locus and demonstrated the crucial role of the WI-1 adhesin in pathogenesis of blastomycosis. Ongoing studies are pointing to a link between phase-specific expression of WI-1 and the observation that transition to yeast cells is essential for the acquisition of pathogenicity by B. dermatitidis. Recombinant attenuated yeast that lack WI-1 are serving as invaluable tools for induction of vaccine resistance and are pointing to new insights about adaptive immunity to B. dermatitidis.     

Blastomyces dermatitidis produces melanin in vitro and during infection

Melanin is made by several important pathogenic fungi and is implicated in the pathogenesis of a number of mycoses. This study investigates whether the thermally dimorphic fungal pathogen Blastomyces dermatitidis produces melanin. Using techniques developed to study melanization in other fungi, we demonstrate that B. dermatitidis conidia and yeast produce melanin in vitro and that yeast cells synthesize melanin or melanin-like pigment in vivo. Melanization reduced susceptibility to amphotericin B, but not to itraconazole or voriconazole. Since melanin is an important virulence factor in other pathogenic fungi, this pigment may affect the pathogenesis of blastomycosis.     

Cell Wall Changes During the Budding Process of Paracoccidioides brasiliensis and Blastomyces dermatitidis

The difference between the budding process of Paracoccidioides brasiliensis and Blastomyces dermatitidis is reported herein. A characteristic feature in P. brasiliensis is that the optical density of the cell wall increases at the site where budding begins and at the neck of the dividing cell, whereas B. dermatitidis does not undergo this alteration. The neck which is formed between the mother and daughter cell at the site of division is much wider in B. dermatitidis than in P. brasiliensis. The bud scar in P. brasiliensis appears as a truncated cone, the top of which is covered only by the inner layer of the cell wall; in comparison, in B. dermatitidis the bud scar exhibits a flattened surface covered by the cell wall. Both fungi show an increase in the number of mitochondria and infoldings of the cytoplasmic membrane at the site of separation, which indicates that at this site there is an increase of metabolic activity.     

Development and Use of a Polyvalent Conjugate to Differentiate Histoplasma capsulatum and Histoplasma duboisii from Other Pathogens

Previous investigations have demonstrated the existence of five Histoplasma capsulatum serotypes. Available specific fluorescent-antibody reagents stain only four of the five serotypes. Antibodies produced against the most complete H. capsulatum serotype were labeled with fluorescein isothiocyanate to develop a reagent specific for H. capsulatum that was reactive with all the known serotypes. The unadsorbed reagent not only stained all the H. capsulatum serotypes, but it also stained cultures of Blastomyces dermatitidis, H. duboisii, several Candida species, and a variety of other fungi. Adsorption of the conjugate with antigens of C. albicans produced a reagent that intensely stained only H. capsulatum, H. duboisii, and B. dermatitidis. Differentiation of B. dermatitidis from the Histoplasma species was accomplished by application of a B. dermatitidis specific fluorescent antibody to antigens positive with the H. capsulatum reagent. At present, differentiation of H. capsulatum from H. duboisii may be accomplished only by animal inoculation. Our data substantiate the antigenic relationships hypothesized earlier, and they indicate that H. capsulatum shares at least two antigens with the other fungi that were studied.     

Effect of soil fungistasis on zoopathogenic fungi

The inhibiting action of west-Siberian soils on spore germination and the growth and development of zoopathogenic fungi such as Emmonsia (Chrysosporium) crescens, Trichophyton mentagrophytes, Beauveria bassiana, Metarrhizium anisopliae, Paecilomyces farinosus, P. fumoso-roseus and Chrysosporium keratinophilum have been studied by the authors. The influence of carbon sources and the root exudates of plants on fungistasis have also been studied.     

Dermatophyte Test Medium: Evaluation with Nondermatophytic Pathogens

The growth and color change produced on dermatophyte test medium (DTM) by 25 strains of zoopathogenic fungi associated with human skin lesions were evaluated quantitatively. DTM only partially suppressed the development of nondermatophytes, with total growth in most instances comparable with that observed with dermatophytes. Whereas all dermatophytes induced a rapid color change on DTM, several nondermatophytic pathogens induced an equally intense and almost as rapid conversion. The rapidity of this color change and the colonial morphology of these forms on DTM could cause their misidentification as dermatophytes under clinical conditions.     

Disinfection of Aerosolized Pathogenic Fungi on Laboratory Surfaces: I. Tissue Phase

The effect of several fungicides on laboratory surfaces contaminated with the tissue phase of aerosolized Blastomyces dermatitidis, Coccidioides immitis, Cryptococcus neoformans, and Histoplasma capsulatum was ascertained. A statistical analysis of the data shows the correlation between fungi, surfaces, time, and concentration of disinfectant. All fungicides were effective at established times and concentrations, and the type of contaminated surface affected the fungicidal efficacy. By interpolating plotted graphs, laboratory personnel may determine, with a given fungicide, the concentration and time required to disinfect instruments, pipettes, gloves, bench tops, and floors contaminated with a tissue phase of pathogenic fungi.     

Coccidioidomycosis and blastomycosis: advances in molecular diagnosis

Clinical isolates of Coccidioides spp. and Blastomyces dermatitidis can be identified by chemiluminescent DNA probes and PCR assays targeting multicopy genes. In fixed tissue samples, cells of the two fungi are specified by in situ hybridization and PCR assays targeting 18S rDNA but sequencing of the products is mandatory. Nested PCR assays targeting genes encoding species- or genus-specific proteins like proline rich antigen of Coccidioides spp. and B. dermatitidis adhesin facilitate amplification of specific DNA from fixed tissue samples. The value of DNA amplification from native specimens of suspected cases of coccidioidomycosis or blastomycosis still needs to be determined.     

Effect of some soybean and bean phytoalexins and related isoflavonoids on growth of Candida albicans

The influence of a series of related isoflavonoids on growth of the zoopathogenic yeast, Candida albicans, in liquid media was studied. The order of effectiveness was identical with that previously found for a range of phytopathogenic fungi. Minimum inhibitory concentration of phaseollinisoflavan, the most active test compound, was 250 microM for 3 days after inoculation. However, C. albicans resumed growth thereafter. Phaseollinisoflavan, 500 microM, did not inhibit but only delayed growth of the yeast. Tolerance of C. albicans to phaseollin and phaseollinisoflavan was based on nondegradative mechanism(s), while tolerance to glyceollin appeared to be based on detoxification.     

The effect of papulacandin B on (1----3)-beta-D-glucan synthetases. A possible relationship between inhibition and enzyme conformation

The antibiotic, papulacandin B, inhibited growth or (1----3)-beta-D-glucan synthetase (or both) in the fungi Saccharomyces cerevisiae, Hansenula anomala, Neurospora crassa, Cryptococcus laurentii, Schizophyllum commune and Wangiella dermatitidis. No effect was observed on Achlya ambisexualis. There was no apparent correlation between the inhibition of growth and that of the synthetase. With most of the fungal extracts, the inhibition of glucan synthetase by papulacandin B became less pronounced as the substrate (UDP-glucose) concentration was decreased. At very low levels of UDP-glucose, with the enzymes from S. cerevisiae and W. dermatitidis, the antibiotic stimulated the activity of glucan synthetase. As further studied with the W. dermatitidis enzyme, those low concentrations of UDP-glucose corresponded to a sigmoidal portion of the rate vs. substrate curve. The sigmoid segment of the curve extended to higher concentrations of UDP-glucose as the temperature was increased. Concomitantly, the range of substrate concentrations at which papulacandin B stimulated the reaction or was noninhibitory was broadened. It is tentatively concluded that glucan synthetase may exist in more than one interconvertible form. The stimulatory effect of papulacandin B is possibly due to preferential binding to the active form of the enzyme. The equilibrium between these forms could be shifted by structural changes in the membrane in which the enzyme is embedded. The lack of correlation between the effects of papulacandin B in whole cells and in extracts is discussed in terms of the variations in membrane structure in the two situations.     

Nests of Marsh harrier (Circus aeruginosus L.) as refuges of potentially phytopathogenic and zoopathogenic fungi

Birds’ nests may be refuges for various species of fungi including that which are potentially phytopathogenic and zoopathogenic. Among the 2449 isolates of fungi obtained from nests of Marsh harriers 96.8% belonged to filamentous fungi. In total, 37 genera were identified from 63 fungi species. Within the mycobiotas of the examined nests populations of fungi which are potentially pathogenic for humans, homoiothermous animals and plants dominated. Among 63 species, 46 (72%) were potentially pathogenic fungi of which 18 species were potentially phytopathogenic and 32 species were pathogenic for homoiothermous animals. Inter alia species of fungi were found in the Marsh harriers nests: Aspergillus fumigatus, Aspergillus flavus, Scopulariopsis brevicaulis, Chrysosporium keratinophilum and Fusarium poae, Fusarium sporotrichioides. In terms of numbers, dominant in Marsh harrier nests were fungi pathogenic to birds, other homoiothermous animals and humans. On that basis it was concluded that Marsh harrier nests are both a source of fungal infections for that species and one of the links in the epidemiological cycle of opportunistic fungi for humans.     

Wangiella dermatitidis isolated from bats in Manaus Brazil

Wangiella dermatitidis was isolated for the first time from the macroscopically asymptomatic internal organs of 2 Phyllostomus discolor, 1 Molossus molossus, 1 Sturnira lilium and 1 Myotis albescens among bats captured in the region of Manaus, Brazil. The dermatiaceous fungi isolated exhibited polymorphism at 25 and 37 degrees C, showing toruloid hyphae, integrated and terminal phialides without collarettes and yeast-like cells. The fungi did not hydrolyze gelatin, casein or starch. Some ecological aspects of bats in relationship to the epidemiology of phaeohyphomycosis are discussed.     

Quantitative Studies of Total Lipids of Pathogenic Fungi

Ten species of dermatophytes and four of the systemic fungi were assayed for total lipids, acetone-soluble fraction, and phospholipid content in different types of cultures. The yeast phase of each of the systemic fungi grown on solid medium exhibited a higher total lipid content than did the mycelial growth in liquid medium, either shake or still. Shake cultures, in all the fungi tested, produced the least lipids. The yeasts were consistently higher also in the acetone-soluble fraction. Histoplasma duboisii in the yeast phase and Microsporum gypseum produced the greatest amount of phospholipid, and Blastomyces dermatitidis in the yeast phase and M. canis produced the largest acetone-soluble fraction among the systemic fungi and dermatophytes, respectively.     

Hydrolysis of vegetable oils and triglycerides by thermotolerant and zoopathogenic species ofAspergillus from Nigerian palm produce

The ability ofAspergillus fumigatus Fres. andAspergillus nidulans (Eidam) Wint obtained from Nigerian palm produce to degrade vegetable oils and triglycerides and the production and activity of their extracellular lipases were studied. Both species readily hydrolysed palm oil and palm kernel oil among others liberating free fatty acids in the process. Good growth with mycelia production of both fungi were also recorded on the triglycerides used as sources of carbon at 37 °C with the best results obtained on palmitic and oleic acids, the predominant fatty acids in palm oil. Extracellular lipases were detected in the culture filtrates of both fungi within 48 h of incubation on an oat-meal chaff medium at 37 °C. Peak enzyme production occurred within the 10-day incubation period. The upases of both fungal species were most active at a pH of 5.6 and a temperature of 45 °C. The best glyceride for assaying the lipase activities of these fungi was trihexanoin while palm oil was a better vegetable oil than the conventional groundnut oil used for the same purpose. Because of the zoopathogenic nature of these fungi, attention is drawn to the potential health risks which their presence on the palm products where they were obtained pose to the consumers.     

Iron gathering by zoopathogenic fungi

Iron is a metal required by most microorganisms and is prominently used in the transfer of electrons during metabolism. The gathering of iron is, then, an essential process and its fulfillment becomes a crucial pathogenetic event for zoopathogenic fungi. Iron is rather unavailable because it occurs on the earth's surface in its insoluble ferric form in oxides and hydroxides. In the infected host iron is bound to proteins such as transferrin and ferritin. Solubilization of ferric iron is the major problem confronting microorganisms. This process is achieved by two major mechanisms: ferric reduction and siderophore utilization. Ferric reductase is frequently accompanied by a copper oxidase transport system. There is one example of direct ferric iron transport apparently without prior reduction. Ferric reduction may also be accomplished by low molecular mass compounds. Some fungi have evolved a process of iron acquisition involving the synthesis of iron-gathering compounds called siderophores. Even those fungi that do not synthesize siderophores have developed permeases for transport of such compounds formed by other organisms. Fungi can also reductively release iron from siderophores and transport the ferrous iron often by the copper oxidase transport system. There is a great diversity of iron-gathering mechanisms expressed by pathogenic fungi and such diversity may be found even in a single species.