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PubMed1990

Utilization by yeasts of D-glucarate, galactarate, and L-tartarate is uncommon and occurs in strains of Cryptococcus and Trichosporon.

In 38 yeast genera tested, utilization of D-glucarate was uncommon, occurring with only 10 strains out of 373. The ability was prominent among Cryptococcus strains, with 8 out of 8 tested being positive, including the pathogen Cryptococcus neoformans. The ability was present also in Trichosporon where 2 out of the 4 strains tested were positive. There was a correlation between ability to utilize D-glucarate, galactarate, L-tartarate, and D-glucuronate. Use of L-ascorbate occurred in more genera than use of D-glucarate, but all strains that grew on D-glucarate grew on L-ascorbate. The utilization of certain hydroxylated carboxylates by strains, mainly found in two genera, is of interest in identifying the catabolic pathways involved, in taxonomic studies, and in developing rapid methods of yeast identification.

Ascorbic Acid↗
PubMedDate not supplied

Cytologic detection of Cryptococcus neoformans in cerebrospinal fluid. Rapid screening methods.

Yeasts of Cryptococcus neoformans in cerebrospinal fluid (CSF) were studied using polarized light and fluorescence microscopy. Ultraviolet-induced fluorescence of wet-fixed, Papanicolaou-stained smears and polarization of air-dried, Diff-Quik or Leishman-stained preparations are complementary methods that allow for rapid screening for and immediate identification of Cryptococcus in cytologic preparations of CSF.

Birefringence↗
PubMed1983

Isolation of Cryptococcus neoformans, Candida albicans and other yeasts from pigeon droppings in Egypt.

Two hundred pigeon droppings, unmixed with soil, and collected from various provinces in lower Egypt (Delta) were cultured to detect yeasts. Yeasts were isolated from 53.5% of the samples; Cryptococcus neoformans was recovered from 30 samples and Candida albicans from 19 samples. Other Cryptococcus and Candida species as well as species of Torulopsis and Rhodotorula were also encountered. Aspergillus fumigatus was isolated on 13 occasions.

Animals↗
PubMed1976

Six-hour pigmentation test for the identification of Cryptococcus neoformans.

Cryptococcus neoformans colonies can be identified within 6 h using paper disks containing caffeic acid and ferric citrate. Indentification is based on the development of a dark brown pigment. Saprophytic Cryptococcus species and common clinically isolated yeasts do not develop the brown color. The concentration of the reagents and the method of storage of the impregnated paper disks are critical for the rapid and specific development of the pigment.

Caffeic Acids↗
PubMed1984

Non-specific immunosuppression by Cryptococcus neoformans infection.

Cryptococcus neoformans-infected animals were found to be immunosuppressed when tested by a variety of assays for immune competence. Primary humoral immune responses and delayed-type hypersensitivity reactions to sheep erythrocytes were suppressed in animals which had been infected for two weeks. Lymphocyte proliferation (LP) assays to sRBC stroma were also significantly diminished at two weeks of infection. Spleen cells of infected mice suppressed the LP response of sRBC immunized, normal mice in vitro. At least a part of the suppression could be attributed to a nylon wool non-adherent cell. Suppressor cells continued to be present in spleen cell suspensions following treatment with anti-T cell serum or anti-immunoglobulin and complement. When infected spleen cells were separated by adherence to plastic, both the adherent and non-adherent fractions exhibited suppressive activity. Incubation of infected spleen cells in tissue culture for 48 hr resulted in the elaboration of soluble immunosuppressive factors into the tissue culture medium. These data indicated that immune suppression in cryptococcosis can occur as a result of infection with Cryptococcus neoformans, and that at least one mechanism involved is the induction of adherent and non-adherent suppressor cells in the spleens of infected mice.

Animals↗
PubMed1990

Polyamine depletion and growth inhibition of Cryptococcus neoformans by alpha-difluoromethylornithine and cyclohexylamine.

The ability of two known inhibitors of polyamine synthesis alpha-difluoromethylornithine (DFMO), an inhibitor of ornithine decarboxylase (ODC), and cyclohexylamine, an inhibitor of spermidine synthase, to inhibit the in vitro growth and polyamine synthesis of clinical isolates of Cryptococcus neoformans was examined. Treatment of C. neoformans with either DFMO or cyclohexylamine resulted in depletion of cellular polyamines and inhibition of growth. Cryptococcus neoformans was shown to lack detectable spermine and to require high concentrations of spermidine, but not putrescine, for growth. The growth inhibition by DFMO and cyclohexylamine was reversed by exogenous polyamines. These findings document the ability of cyclohexylamine and DFMO to inhibit polyamine synthesis and growth in clinically important isolates of C. neoformans.

Cryptococcus neoformans↗
PubMed1991

Immunity to a pulmonary Cryptococcus neoformans infection requires both CD4+ and CD8+ T cells.

The role of CD4+ and CD8+ T cells in mediating pulmonary clearance of a cryptococcal infection was investigated. Intratracheal inoculation of BALB/c and C.B-17 mice with a moderately virulent strain of Cryptococcus neoformans (52D) resulted in a pulmonary infection, which was cleared by a T cell-dependent mechanism. During this clearance, there was a significant influx of both CD4+ and CD8+ T cells into the lungs. Depletion of CD4+ T cells by injections of CD4-specific monoclonal antibody (mAb) prevented pulmonary clearance and also resulted in significant colonization of the brain and spleen of infected mice. CD4 depletion did not prevent the influx of CD8+ T cells into the lungs. Surprisingly, depletion of CD8+ T cells by mAb also ablated pulmonary clearance. CD8-depleted mice also had a small but significant increase in brain and spleen colony-forming unit compared to control mice by the end of the study. CD4+ T cell pulmonary influx was independent of the presence of CD8+ T cells. The lungs of T cell-depleted mice were examined histologically. CD4+ and CD8+ T cells each mediated a degree of inflammatory influx seen in the lungs of infected mice and raised the possibility that CD4+ and CD8+ T cells may synergize to generate the inflammatory response in the lungs. Numerous phagocytized but intact cryptococci were seen in the inflammatory foci of CD8-depleted mice but not in control or CD4-depleted mice. We propose that CD4+ T cells may recruit and activate effector phagocytes while CD8+ T cells predominantly function to lyse cryptococcus-laden unactivated phagocytes similar to the function of CD8+ T cells during listeria and mycobacteria infections.

Animals↗
PubMed1992

CD4+ T cells cause multinucleated giant cells to form around Cryptococcus neoformans and confine the yeast within the primary site of infection in the respiratory tract.

The possible mechanisms by which CD4+ T cells prevent the dissemination of Cryptococcus neoformans from the primary site of infection in the respiratory tract were examined. It was found that even before fungicidal mechanisms are fully induced in the lungs, the host generates a CD4+ T cell-dependent inflammatory response that sequesters yeast within the pulmonary alveoli. This confinement is evident histopathologically and demonstrable objectively as a rapid decline in the ability to dislodge yeast from the lungs by bronchopulmonary lavage. One striking component of this response is the enclosure of cryptococci within multinucleated giant cells in granulomas. Studies in severe combined immunodeficient mice that were engrafted with selected lymphocyte subpopulations show that B cells, and hence anti-Cryptococcus antibodies, are not necessary for the CD4+ T cell-dependent responses that isolate and subsequently destroy this opportunistic pathogen in the lung parenchyma.

Animals↗
PubMedDate not supplied

The ecology of Cryptococcus neoformans and the epidemiology of cryptococcosis.

The ecology of Cryptococcus neoformans and the epidemiology of cryptococcosis are reviewed. Two varieties of C. neoformans have been recognized. C. neoformans variety neoformans has been found in nature worldwide, primarily in association with bird droppings, although nonavian sources have also been found. Most cases of human cryptococcosis are caused by this variety. C. neoformans var. gattii has recently been isolated in nature in association with Eucalyptus trees. Infections caused by this variety occur mainly in tropical and subtropical regions. Because exposure to C. neoformans is probably common and clinically apparent cases of cryptococcosis in healthy hosts are rare, it is presumed that most people can mount adequate host defenses upon exposure to the organism. At least 5%-10% of patients with AIDS become infected with Cryptococcus; the epidemiology of this infection is different in many respects from that seen in patients without AIDS.

Acquired Immunodeficiency Syndrome↗
PubMed1988

Taxonomic relationships of Cryptococcus and Tremella based on fatty acid composition and other phenotypic characters.

The cellular long-chain fatty acids present in 33 strains, representing 15 species of Cryptococcus, and 4 species of Tremella, were determined by gas chromatography. According to the relative amounts of fatty acid methyl esters, the Cryptococcus species studied were divided into four main groups. Possible relationships between species representing the two genera are presented in a new model, where cellular long-chain fatty acid compositions and other phenotypic characteristics are included.

Chromatography, Gas↗
PubMed1966

Medium for selective isolation of Cryptococcus neoformans.

A medium has been developed that permits the selective recovery of Cryptococcus neoformans from heavily contaminated materials. It employs creatinine as a nitrogen source, diphenyl (C(6)H(5)C(6)H(5)) and chloramphenicol as mold and bacterial inhibitors, and Guizotia abyssinica seed extract as a specific color marker. The medium has proved to be effective in the direct isolation of Cryptococcus neoformans from pigeon nests and from the air.

Biphenyl Compounds↗
PubMed1968

Fluorescent-antibody reagent for the identification of Cryptococcus neoformans.

A sensitive and diagnostically applicable conjugate for the rapid and accurate detection and identification of Cryptococcus neoformans has been developed. C. neoformans rabbit antisera were produced by 14 daily intravenous injections of 36 million cells for a total dosage of approximately 500 million cells. Cross-staining reactions with species of Cryptococcus other than C. neoformans, as well as with Candida species, were eliminated by adsorption of the C. neoformans conjugate with cells of C. diffluens and C. krusei.

Adsorption↗
PubMed1971

Pigment formation for differentiating Cryptococcus neoformans from Candida albicans.

When 2,3- or 3,4-dihydroxybenzoic acid, 3,4-dihydroxyphenylalanine, and 3,4-dihydroxycinnamic acid are added to growth media, they are converted to a characteristic brown pigment by Cryptococcus neoformans. This pigment formation has hitherto been encountered only when this microorganism was cultivated on media containing Guizotia abyssinica seed. This phenomenon can be used for differentiating Cryptococcus neoformans from Candida albicans. Possible precursors of these o-diphenols (quinic acid, aromatic monohydroxy acids, or tyrosine) do not give rise to the brown pigmentation.

Agar↗
PubMed1992

Virulence, serotype, and molecular characteristics of environmental strains of Cryptococcus neoformans var. gattii.

Four strains of Cryptococcus neoformans var. gattii originating from Eucalyptus camaldulensis, three from Australia and one from San Francisco, were tested for their serotype, virulence for mice, and a number of genetic and molecular characteristics. All were found to be serotype B and showed significantly higher virulence for mice than did the type strains of C. neoformans var. gattii and Filobasidiella neoformans var. bacillispora, which were obtained from human cryptococcosis cases. Electrophoretic karyotypes of the strains from Australia were identical, although they were collected from sites at least 15 to 500 km apart. The electrophoretic karyotype of the strain from San Francisco was the same as that of the Australian isolates except for the mobility of one chromosome. On the contrary, no two isolates of serotype B (of a total of 11) from clinical sources were the same, regardless of their geographic origin. Furthermore, none of the clinical isolates showed a chromosomal banding pattern identical to that of Eucalyptus-originated strains. The Eucalyptus-originated strains failed to form dikaryons when crossed with the tester strains of the two varieties of F. neoformans. Hybridization analysis with a nucleic acid probe (AccuProbe C. neoformans Culture Confirmation Test; Gen-Probe Inc., San Diego, Calif.), however, showed signals of equal intensity for clinical strains and the Eucalyptus-originated strains. Various fungi phylogenetically related to C. neoformans, including a phenol oxidase-positive strain of Cryptococcus laurentii obtained from E. camaldulensis, were negative in the nucleic acid hybridization test. These observations confirm that, in spite of karyotypic differences and the lack of dikaryon formation with the tester strains of F. neoformans, Eucalyptus-originated C. neoformans var. gattii is the same organism as those isolated from cases of human infection. Furthermore, the C. neoformans culture confirmation test using a commercial nucleic acid probe is specific for C. neoformans.

Animals↗
PubMed1970

Rhodotorulic acid from species of Leucosporidium, Rhodosporidium, Rhodotorula, Sporidiobolus, and Sporobolomyces, and a new alanine-containing ferrichrome from Cryptococcus melibiosum.

An examination of 142 strains within 19 genera of yeasts and yeastlike organisms for formation of hydroxamic acids in low-iron culture showed production of hydroxamates by two unclassified strains and by 52 strains among the genera Aessosporon (3 of 3 strains), Cryptococcus (1 of 43), Leucosporidium (3 of 11), Rhodosporidium (4 of 4), Rhodotorula (27 of 39), Sporidiobolus (2 of 2), and Sporobolomyces (12 of 13). Crystalline rhodotorulic acid was isolated in amounts sufficient to account for most or all of the measured hydroxamate in culture supernatants of 16 strains representative of the five last-mentioned hydroxamate-producing genera. A new alanine-containing ferrichrome was isolated from one strain of Cryptococcus melibiosum. Rhodotorulic acid was a major metabolic product of many of the positive strains when grown in low-iron media, and iron was shown to repress its synthesis and excretion into the culture medium. The taxonomic significance of production of hydroxamic acids is described in connection with the position of these yeast species in the subclass Heterobasidiomycetidae.

Alanine↗
PubMed1976

Production of extracellular ribonuclease by yeasts and yeastlike fungi, and its repression by orthophosphate in species of Cryptococcus and Tremella.

A strain of Cryptococcus laurentii and a haploid isolate of Tremella foliacea were shown to produce orthophosphate-repressible ribonuclease in liquid culture. Addition of as little as 1 mM K2HPO4, pH 7.0, completely repressed enzyme production by both fungi. The orthophosphate-repressible enzyme was not produced by other species of the two genera tested. These results, together with other findings, suggest a close phylogenetic relationship between Cryptococcus laurentii and Tremella foliacea. The ability of other yeasts and yeastlike fungi to hydrolyze ribonucleic acid in a solid test medium was assessed. Based on the limited number of organisms available for study, extracellular ribonuclease activity was found in species having close affinity to the Basidiomycetes and in yeasts classified in the ascomycetous genera, Endomycopsis, Hansenula, and Kluyveromyces. Other ascomycetous yeasts did not exhibit extracellular ribonuclease.

Ascomycota↗
PubMed1990

Rapid genetic identification and mapping of enzymatically amplified ribosomal DNA from several Cryptococcus species.

Detailed restriction analyses of many samples often require substantial amounts of time and effort for DNA extraction, restriction digests, Southern blotting, and hybridization. We describe a novel approach that uses the polymerase chain reaction (PCR) for rapid simplified restriction typing and mapping of DNA from many different isolates. DNA fragments up to 2 kilobase pairs in length were efficiently amplified from crude DNA samples of several pathogenic Cryptococcus species, including C. neoformans, C. albidus, C. laurentii, and C. uniguttulatus. Digestion and electrophoresis of the PCR products by using frequent-cutting restriction enzymes produced complex restriction phenotypes (fingerprints) that were often unique for each strain or species. We used the PCR to amplify and analyze restriction pattern variation within three major portions of the ribosomal DNA (rDNA) repeats from these fungi. Detailed mapping of many restriction sites within the rDNA locus was determined by fingerprint analysis of progressively larger PCR fragments sharing a common primer site at one end. As judged by PCR fingerprints, the rDNA of 19 C. neoformans isolates showed no variation for four restriction enzymes that we surveyed. Other Cryptococcus spp. showed varying levels of restriction pattern variation within their rDNAs and were shown to be genetically distinct from C. neoformans. The PCR primers used in this study have also been successfully applied for amplification of rDNAs from other pathogenic and nonpathogenic fungi, including Candida spp., and ought to have wide applicability for clinical detection and other studies.

Base Sequence↗
PubMed1966

Epidemiological and immunological studies of Cryptococcus neoformans.

Walter, Jinks E. (University of Pittsburgh, Pittsburgh, Pa.), and Robert W. Atchison. Epidemiological and immunological studies of Cryptococcus neoformans. J. Bacteriol. 92:82-87. 1966.-The complement-fixation fluorescent-antibody test provided a means of differentiating between antibodies of Cryptococcus neoformans and Candida albicans. The test was applied to the sera of 134 pigeon fanciers for detection of antibodies to C. neoformans only. About 22% were positive as compared with 3% of a control group composed of 36 non-pigeon breeders. Positive reactions were observed only with C. neoformans types A and B cells. It was concluded that the pigeon fanciers had presumably been infected previously with C. neoformans type A or type B. Moreover, 48 of 49 isolates of C. neoformans cultured from the pigeon habitats of 72 fanciers studied were serotype A. These findings would seem to substantiate the hypothesis that pigeon habitats serve as reservoirs for human infections, and also that subclinical cryptococcosis is more prevalent than is realized.

Animals↗