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Sample records for cryptococcus gattii fungemia

  1. Cryptococcus gattii fungemia: report of a case with lung and brain lesions mimicking radiological features of malignancy Fungemia por Cryptococcus gattii: relato de um caso com lesões cerebrais e pulmonares nos achados radiológicos mimetizando câncer

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    Flávio de Mattos Oliveira

    2007-08-01

    Full Text Available A 64-year-old apparently immunocompetent white man developed lung and brain lesions of disseminated cryptococcosis. The radiologic features mimicked those of lung cancer metastatic to the central nervous system. C. gattii was recovered from cultures of bronchoalveolar lavage fluid, brain biopsy, and blood. The same fungus was recovered from pulmonary and brain specimens at autopsy. Serum and cerebrospinal fluid cryptococcal antigen tests were diagnostic in our case and should be included in the diagnostic evaluation of unexplained pulmonary and cerebral lesions. A literature search showed few reports of fungemia by this species of Cryptococcus, contrasting to C. neoformans.Homem branco de 64 anos, aparentemente imunocompetente, desenvolveu lesões pulmonares e cerebrais por criptococose disseminada. Os achados radiológicos foram similares àqueles encontrados em pacientes com câncer de pulmão e metástase no sistema nervoso central. C. gattii foi isolado de cultivos de lavado broncoalveolar, biópsia cerebral e sangue. O mesmo fungo foi encontrado em fragmentos pulmonares e cerebrais obtidos da autópsia. Testes de antígeno no soro e no líquido cefalorraquidiano foram diagnóstico no nosso caso e devem ser incluídos na avaliação diagnóstica de lesões pulmonares e cerebrais indefinidas. Pesquisa na literatura mostrou poucos relatos de fungemia por esta espécie de Cryptococcus, contrastando com C. neoformans.

  2. Unisexual reproduction of Cryptococcus gattii.

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    Sujal S Phadke

    Full Text Available Cryptococcus gattii is a basidiomycetous human fungal pathogen that typically causes infection in tropical and subtropical regions and is responsible for an ongoing outbreak in immunocompetent individuals on Vancouver Island and in the Pacific Northwest of the US. Pathogenesis of this species may be linked to its sexual cycle that generates infectious propagules called basidiospores. A marked predominance of only one mating type (α in clinical and environmental isolates suggests that a-α opposite-sex reproduction may be infrequent or geographically restricted, raising the possibility of an alternative unisexual cycle involving cells of only α mating type, as discovered previously in the related pathogenic species Cryptococcus neoformans. Here we report observation of hallmark features of unisexual reproduction in a clinical isolate of C. gattii (isolate 97/433 and describe genetic and environmental factors conducive to this sexual cycle. Our results are consistent with population genetic evidence of recombination in the largely unisexual populations of C. gattii and provide a useful genetic model for understanding how novel modes of sexual reproduction may contribute to evolution and virulence in this species.

  3. Diversity of the Cryptococcus neoformans-Cryptococcus gattii species complex.

    NARCIS (Netherlands)

    Bovers, M.; Hagen, F.; Boekhout, T.

    2008-01-01

    More than 110 years of study of the Cryptococcus neoformans and Cryptococcus gattii species complex has resulted in an enormous accumulation of fundamental and applied biological and clinical knowledge. Recent developments in our understanding of the diversity within the species complex are

  4. Cryptococcus gattii and Cryptococcus neoformans - Cosmopolitans on the move

    NARCIS (Netherlands)

    Hagen, F.

    2011-01-01

    The fungal pathogens Cryptococcus neoformans and Cryptococcus gattii are the leading cause of fungal meningitis worldwide. Until three decades ago both pathogenic yeast species were, however, rarely encountered. The onset of the HIV-pandemic during the early 1980s showed the deadly potential of the

  5. Equine nasopharyngeal cryptococcoma due to Cryptococcus gattii

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    Raquel Aparecida Sales da Cruz

    2017-08-01

    Full Text Available ABSTRACT: Cryptococcus gattii is often associated with pulmonary and systemic infections in humans and animals. In this research we report a case of nasopharyngeal cryptococoma caused by C. gatti in an equine. A 10-year-old mare presented a mass obstructing the oropharynx. Macroscopically the mass was asymmetric, and was attached to the ethmoidal sinuses and obstructed the oropharynx. Histopathological examination of the mass revealed multiple yeast cells ranging from spherical to oval, 4-8μm in diameter, with some of them showing narrow base polar budding. Cryptococcus gattii growth in mycological culture (Sabouraud Dextrose Agar and was L-canavanine-glycine-bromothymol blue Agar positive. The molecular identification confirmed the isolate as C. gattii by means of the amplification of universal primers. C. gattii is considered an emerging fungal agent, as it affects human and animals and does not respond efficiently to commonly established treatments.

  6. Cryptococcus gattii : A dilemma in diagnosis and treatment in sub ...

    African Journals Online (AJOL)

    Sub-Saharan Africa contributes at least 70% of the global cryptococcal meningoencephalitis cases each year and the majority of cases are caused by the Cryptococcus neoformans species. We present a case of meningoencephalitis due to Cryptococcus gattii in an 18 year old apparently immunocompetent male patient ...

  7. Fundamental niche prediction of the pathogenicyeasts Cryptococcus neoformans and Cryptococcus gattii in Europe

    NARCIS (Netherlands)

    Cogliati, M.; Puccianti, E.; Montagna, M.T.; De Donno, A.; Susever, S.; Ergin, C.; Velegraki, A.; Ellabib, M.S.; Nardoni, S.; Macci, C.; Trovato, L.; Dipineto, L.; Rickerts, V.; Akcaglar, S.; Mlinaric-Missoni, E.; Bertout, S.; Venca, A.C.F.; Sampaio, A.C.; Criseo, G.; Ranque, S.; Cerikcioglu, N.; Marchese, A.; Vezzulli, L.; Ilkit, M.; Desnos-Ollivier, M.; Pasquale, V.; Polacheck, I.; Scopa, A.; Meyer, W.; Ferreira-Paim, K.; Hagen, F.; Boekhout, T.; Dromer, F.; Varma, A.; Kwon-Chung, K.J.; Inacio, J.; Colom, M.F.

    2017-01-01

    Fundamental niche prediction of Cryptococcus neoformans and Cryptococcus gattii in Europe is an important tool to understand where these pathogenic yeasts have a high probability to survive in the environment and therefore to identify the areas with high risk of infection. In this study, occurrence

  8. Is Climate influencing Cryptococcus gattii on Vancouver Island?

    Centers for Disease Control (CDC) Podcasts

    2016-03-24

    Dr. Christopher Uejio, Department of Geography and Program in Public Health, Florida State University, discusses Cryptococcus gattii on Vancouver Island.  Created: 3/24/2016 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 3/24/2016.

  9. Identification of genes expressed by Cryptococcus gattii during iron deprivation

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    Daphine Ariadne Jesus de Paula

    2014-09-01

    Full Text Available Cryptococcus neoformans and C. gattii are pathogenic yeasts that cause life-threatening diseases in humans and animals. Iron is an essential nutrient for virtually every organism as it functions as a cofactor in numerous essential enzymatic reactions. In the literature, the competition for iron between microbes and mammalian hosts during infection is well documented. In this study, we used representational difference analysis (RDA in order to gain a better understanding of how C. gattii responds to iron starvation. A total of 15 and 29 genes were identified as having altered expression levels due to iron depletion after 3 h and 12 h, respectively. Of these, eight genes were identified in both libraries. The transcripts were related to many biological processes, such as cell cycle, ergosterol metabolism, cell wall organization, transportation, translation, cell respiration and the stress response. These data suggest a remodeling of C. gattii metabolism during conditions of iron deprivation.

  10. Long-term survival of Cryptococcus neoformans and Cryptococcus gattii in stored environmental samples from Colombia.

    Science.gov (United States)

    Escandón, Patricia; Castañeda, Elizabeth

    2015-01-01

    Both Cryptococcus neoformans and Cryptococcus gattii have been isolated from a variety of environmental sources in Colombia. To determine the viability of C. neoformans/C. gattii isolates in stored soil samples, filtrates and bird droppings from which these yeasts were previously recovered. A total of 964 samples collected between 2003 and 2009, and kept at room temperature were processed. From them, 653 samples were from trees decaying wood, 274 from soil filtrates and 37 from bird droppings. When C. neoformans or C. gattii were recovered, the molecular type of each isolate was established by PCR fingerprinting using the single primer (GTG)5. Among the processed samples, 161 isolates were recovered. From those, 81 (50.3%) corresponded to C. gattii recovered from decaying wood of Eucalyptus spp., Corymbia ficifolia, Terminalia catappa and Ficus spp. trees, and 80 (49.7%) corresponded to C. neoformans recovered from Ficus spp. and eucalyptus trees, as well as from bird droppings. The most prevalent molecular type among the C. gattii and C. neoformans isolates was VGII and VNI, respectively. The re-isolation of C. neoformans/C. gattii from 10-year stored samples suggests that these yeasts are able to keep viable in naturally colonized samples. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  11. Epidemiologische trends in cryptokokkose : De Cryptococcus gattii-uitbraak in Canada

    NARCIS (Netherlands)

    Hagen, F; Boekhout, T

    2006-01-01

    Cryptococcosis is, when untreated, a fatal disease, which in the Netherlands is mainly caused by the basidiomycetous yeast Cryptococcus neoformans, and in more rare cases by Cryptococcus gattii. Infections with C. gattii occur almost only in immunocompetent individuals, while C. neoformans has a

  12. Mitochondrial DNA inheritance in the human fungal pathogen Cryptococcus gattii.

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    Wang, Zixuan; Wilson, Amanda; Xu, Jianping

    2015-02-01

    The inheritance of mitochondrial DNA (mtDNA) is predominantly uniparental in most sexual eukaryotes. In this study, we examined the mitochondrial inheritance pattern of Cryptococcus gattii, a basidiomycetous yeast responsible for the recent and ongoing outbreak of cryptococcal infections in the US Pacific Northwest and British Columbia (especially Vancouver Island) in Canada. Using molecular markers, we analyzed the inheritance of mtDNA in 14 crosses between strains within and between divergent lineages in C. gattii. Consistent with results from recent studies, our analyses identified significant variations in mtDNA inheritance patterns among strains and crosses, ranging from strictly uniparental to biparental. For two of the crosses that showed uniparental mitochondrial inheritance in standard laboratory conditions, we further investigated the effects of the following environmental variables on mtDNA inheritance: UV exposure, temperature, and treatments with the methylation inhibitor 5-aza-2'-deoxycytidine and with the ubiquitination inhibitor ammonium chloride. Interestingly, one of these crosses showed no response to these environmental variables while the other exhibited diverse patterns ranging from complete uniparental inheritance of the MATa parent mtDNA, to biparental inheritance, and to a significant bias toward inheritance of the MATα parental mtDNA. Our results indicate that mtDNA inheritance in C. gattii differs from that in its closely related species Cryptococcus neoformans. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Fundamental niche prediction of the pathogenic yeasts Cryptococcus neoformans and Cryptococcus gattii in Europe.

    Science.gov (United States)

    Cogliati, Massimo; Puccianti, Erika; Montagna, Maria T; De Donno, Antonella; Susever, Serdar; Ergin, Cagri; Velegraki, Aristea; Ellabib, Mohamed S; Nardoni, Simona; Macci, Cristina; Trovato, Laura; Dipineto, Ludovico; Rickerts, Volker; Akcaglar, Sevim; Mlinaric-Missoni, Emilija; Bertout, Sebastien; Vencà, Ana C F; Sampaio, Ana C; Criseo, Giuseppe; Ranque, Stéphane; Çerikçioğlu, Nilgün; Marchese, Anna; Vezzulli, Luigi; Ilkit, Macit; Desnos-Ollivier, Marie; Pasquale, Vincenzo; Polacheck, Itzhack; Scopa, Antonio; Meyer, Wieland; Ferreira-Paim, Kennio; Hagen, Ferry; Boekhout, Teun; Dromer, Françoise; Varma, Ashok; Kwon-Chung, Kyung J; Inácio, Joäo; Colom, Maria F

    2017-10-01

    Fundamental niche prediction of Cryptococcus neoformans and Cryptococcus gattii in Europe is an important tool to understand where these pathogenic yeasts have a high probability to survive in the environment and therefore to identify the areas with high risk of infection. In this study, occurrence data for C. neoformans and C. gattii were compared by MaxEnt software with several bioclimatic conditions as well as with soil characteristics and land use. The results showed that C. gattii distribution can be predicted with high probability along the Mediterranean coast. The analysis of variables showed that its distribution is limited by low temperatures during the coldest season, and by heavy precipitations in the driest season. C. neoformans var. grubii is able to colonize the same areas of C. gattii but is more tolerant to cold winter temperatures and summer precipitations. In contrast, the C. neoformans var. neoformans map was completely different. The best conditions for its survival were displayed in sub-continental areas and not along the Mediterranean coasts. In conclusion, we produced for the first time detailed prediction maps of the species and varieties of the C. neoformans and C. gattii species complex in Europe and Mediterranean area. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

  14. Pigment Production on L-Tryptophan Medium by Cryptococcus gattii and Cryptococcus neoformans

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    Chaskes, Stuart; Cammer, Michael; Nieves, Edward; Casadevall, Arturo

    2014-01-01

    In recent years strains previously grouped within Cryptococcus neoformans have been divided into two species C. neoformans and C. gattii, with Cryptococcus neoformans comprising serotypes A, D, and AD and C. gattii comprising serotypes B and C. Cryptococcus neoformans have also been subdivided into two varieties C. neoformans var. grubii, serotype A, and C. neoformans var. neoformans, serotype D. We analyzed the growth and pigment production characteristics of 139 strains of Cryptococcus spp. in L-tryptophan containing media. Nearly all strains of Cryptococcus, including each variety and serotype tested produced a pink water-soluble pigment (molecular weight of 535.2 Da) from L-tryptophan. Consequently, the partial separation of the species was based on whether the pink pigment was secreted into the medium (extracellular) or retained as an intracellular pigment. On L-tryptophan medium C. neoformans var. grubii and serotype AD produced a pink extracellular pigment. In contrast, for C. gattii, the pink pigment was localized intracellularly and masked by heavy production of brown pigments. Pigment production by C. neoformans var. neoformans was variable with some strains producing the pink extracellular pigment and others retained the pink pigment intracellularly. The pink intracellular pigment produced by strains of C. neoformans var. neoformans was masked by production of brown pigments. Cryptococcus laccase mutants failed to produce pigments from L-tryptophan. This is the first report that the enzyme laccase is involved in tryptophan metabolism. Prior to this report Cryptococcus laccase produced melanin or melanin like-pigments from heterocyclic compounds that contained ortho or para diphenols, diaminobenzenes and aminophenol compounds. The pigments produced from L-tryptophan were not melanin. PMID:24736553

  15. Pigment production on L-tryptophan medium by Cryptococcus gattii and Cryptococcus neoformans.

    Science.gov (United States)

    Chaskes, Stuart; Cammer, Michael; Nieves, Edward; Casadevall, Arturo

    2014-01-01

    In recent years strains previously grouped within Cryptococcus neoformans have been divided into two species C. neoformans and C. gattii, with Cryptococcus neoformans comprising serotypes A, D, and AD and C. gattii comprising serotypes B and C. Cryptococcus neoformans have also been subdivided into two varieties C. neoformans var. grubii, serotype A, and C. neoformans var. neoformans, serotype D. We analyzed the growth and pigment production characteristics of 139 strains of Cryptococcus spp. in L-tryptophan containing media. Nearly all strains of Cryptococcus, including each variety and serotype tested produced a pink water-soluble pigment (molecular weight of 535.2 Da) from L-tryptophan. Consequently, the partial separation of the species was based on whether the pink pigment was secreted into the medium (extracellular) or retained as an intracellular pigment. On L-tryptophan medium C. neoformans var. grubii and serotype AD produced a pink extracellular pigment. In contrast, for C. gattii, the pink pigment was localized intracellularly and masked by heavy production of brown pigments. Pigment production by C. neoformans var. neoformans was variable with some strains producing the pink extracellular pigment and others retained the pink pigment intracellularly. The pink intracellular pigment produced by strains of C. neoformans var. neoformans was masked by production of brown pigments. Cryptococcus laccase mutants failed to produce pigments from L-tryptophan. This is the first report that the enzyme laccase is involved in tryptophan metabolism. Prior to this report Cryptococcus laccase produced melanin or melanin like-pigments from heterocyclic compounds that contained ortho or para diphenols, diaminobenzenes and aminophenol compounds. The pigments produced from L-tryptophan were not melanin.

  16. Pigment production on L-tryptophan medium by Cryptococcus gattii and Cryptococcus neoformans.

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    Stuart Chaskes

    Full Text Available In recent years strains previously grouped within Cryptococcus neoformans have been divided into two species C. neoformans and C. gattii, with Cryptococcus neoformans comprising serotypes A, D, and AD and C. gattii comprising serotypes B and C. Cryptococcus neoformans have also been subdivided into two varieties C. neoformans var. grubii, serotype A, and C. neoformans var. neoformans, serotype D. We analyzed the growth and pigment production characteristics of 139 strains of Cryptococcus spp. in L-tryptophan containing media. Nearly all strains of Cryptococcus, including each variety and serotype tested produced a pink water-soluble pigment (molecular weight of 535.2 Da from L-tryptophan. Consequently, the partial separation of the species was based on whether the pink pigment was secreted into the medium (extracellular or retained as an intracellular pigment. On L-tryptophan medium C. neoformans var. grubii and serotype AD produced a pink extracellular pigment. In contrast, for C. gattii, the pink pigment was localized intracellularly and masked by heavy production of brown pigments. Pigment production by C. neoformans var. neoformans was variable with some strains producing the pink extracellular pigment and others retained the pink pigment intracellularly. The pink intracellular pigment produced by strains of C. neoformans var. neoformans was masked by production of brown pigments. Cryptococcus laccase mutants failed to produce pigments from L-tryptophan. This is the first report that the enzyme laccase is involved in tryptophan metabolism. Prior to this report Cryptococcus laccase produced melanin or melanin like-pigments from heterocyclic compounds that contained ortho or para diphenols, diaminobenzenes and aminophenol compounds. The pigments produced from L-tryptophan were not melanin.

  17. Ancient dispersal of the human fungal pathogen Cryptococcus gattii from the Amazon rainforest

    NARCIS (Netherlands)

    Hagen, F.; Ceresini, P.C.; Polacheck, I.; Ma, H.; van Nieuwerburgh, F.; Gabaldon, T.; Kagan, S.; Pursall, E.R.; Hoogveld, H.L.; van Iersel, L.J.; Klau, G.W.; Kelk, S.M.; Stougie, L.; Bartlett, K.H.; Voelz, K.; Pryszcz, L.P.; Castaneda, E.; Lazera, M.; Meyer, W.; Deforce, D.; Meis, J.F.G.M.; May, R.C.; Klaassen, C.H.W.; Boekhout, T.

    2013-01-01

    Over the past two decades, several fungal outbreaks have occurred, including the high-profile 'Vancouver Island' and 'Pacific Northwest' outbreaks, caused by Cryptococcus gattii, which has affected hundreds of otherwise healthy humans and animals. Over the same time period, C. gattii was the cause

  18. Ancient dispersal of the human fungal pathogen Cryptococcus gattii from the Amazon rainforest.

    NARCIS (Netherlands)

    Hagen, F.; Ceresini, P.C.; Polacheck, I.; Ma, H.; Nieuwerburgh, F. van; Gabaldón, T.; Kagan, S.; Pursall, E.R.; Hoogveld, H.L.; Iersel, L.J. van; Klau, G.W.; Kelk, S.M.; Stougie, L.; Bartlett, K.H.; Voelz, K.; Pryszcz, L.P.; Castañeda, E.; Lazera, M.; Meyer, W.; Deforce, D.; Meis, J.F.G.M.; May, R.C.; Klaassen, C.H.; Boekhout, T.

    2013-01-01

    Over the past two decades, several fungal outbreaks have occurred, including the high-profile 'Vancouver Island' and 'Pacific Northwest' outbreaks, caused by Cryptococcus gattii, which has affected hundreds of otherwise healthy humans and animals. Over the same time period, C. gattii was the cause

  19. Meningitis secondary to Cryptococcus gattii,an emerging pathogen affecting immunocompetent hosts

    Institute of Scientific and Technical Information of China (English)

    Laura Cookman; Maria Hugi

    2013-01-01

    BACKGROUND:Meningitis continues to be one of the most important infections diagnosed and treated by emergency physicians.Despite the advent of anti-infective therapy,meningitis carries a mortality rate of 20%-40%.In this study,we describe the first reported emergency medicine case of meningitis associated with Cryptococcus gattii to alert providers of this insidious,emerging global pathogen infecting immunocompetent individuals.METHODS:We provided a case report and accompanying review of the literature.A MEDLINE search for the term Cryptoccocus gattii was performed to obtain background information on Cryptococcus gattii.RESULTS:After two months of hospitalization,the patient was eventually discharged neurologically intact except for a continued mild bilateral hearing deficit.CONCLUSION:Cryptococcus gattii is an emerging world pathogen,which affects otherwise healthy,immunocompetent patients and requires timely identification and treatment in order to prevent severe neurological sequelae.

  20. Clinical and microbiological characteristics of cryptococcosis in Singapore: predominance of Cryptococcus neoformans compared with Cryptococcus gattii.

    Science.gov (United States)

    Chan, Monica; Lye, David; Win, Mar Kyaw; Chow, Angela; Barkham, Tim

    2014-09-01

    To describe the clinical features, treatments, outcomes, and subtype prevalence of cryptococcosis in Singapore. All patients with laboratory confirmed cryptococcal infections admitted from 1999 to 2007 to a teaching hospital in Singapore were reviewed retrospectively. Identification and molecular types of Cryptococcus neoformans variants and Cryptococcus gattii were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Serotypes were inferred with a multiplex PCR method. Of 62 patients with cryptococcosis, C. neoformans var. grubii was the predominant subtype (in 95%), affecting mainly immunocompromised hosts (91%) with HIV infection (80%). Patients with HIV were younger (median age 36.5 vs. 49.5 years, p=0.006) and less likely to present with an altered mental status (14% vs. 50%, p=0.013). In contrast, delayed treatment (median 7 days vs. 2 days, p=0.03), pulmonary involvement (58% vs. 14%, p=0.03), and initial treatment with fluconazole (25% vs. 2%, p=0.02) were more common in HIV-negative patients. C. gattii was uncommon, affecting only three patients, all of whom were immunocompetent and had disseminated disease with pulmonary and neurological involvement. All C. gattii were RFLP type VG II, serotype B and all C. neoformans var. grubii were RFLP type VN I, serotype A, except for one that was RFLP type VN II. C. neoformans var. grubii, subtype VN I, was the predominant subtype in Singapore, infecting younger, mainly immunocompromised hosts with HIV. C. gattii was uncommon, causing pulmonary manifestations in older, immunocompetent patients and were RFLP type VG II. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

  1. Vaccine-mediated immune responses to experimental pulmonary Cryptococcus gattii infection in mice.

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    Ashok K Chaturvedi

    Full Text Available Cryptococcus gattii is a fungal pathogen that can cause life-threatening respiratory and disseminated infections in immune-competent and immune-suppressed individuals. Currently, there are no standardized vaccines against cryptococcosis in humans, underlying an urgent need for effective therapies and/or vaccines. In this study, we evaluated the efficacy of intranasal immunization with C. gattii cell wall associated (CW and/or cytoplasmic (CP protein preparations to induce protection against experimental pulmonary C. gattii infection in mice. BALB/c mice immunized with C. gattii CW and/or CP protein preparations exhibited a significant reduction in pulmonary fungal burden and prolonged survival following pulmonary challenge with C. gattii. Protection was associated with significantly increased pro-inflammatory and Th1-type cytokine recall responses, in vitro and increased C. gattii-specific antibody production in immunized mice challenged with C. gattii. A number of immunodominant proteins were identified following immunoblot analysis of C. gattii CW and CP protein preparations using sera from immunized mice. Immunization with a combined CW and CP protein preparation resulted in an early increase in pulmonary T cell infiltrates following challenge with C. gattii. Overall, our studies show that C. gattii CW and CP protein preparations contain antigens that may be included in a subunit vaccine to induce prolonged protection against pulmonary C. gattii infection.

  2. Native trees of the Northeast Argentine: natural hosts of the Cryptococcus neoformans-Cryptococcus gattii species complex.

    Science.gov (United States)

    Cattana, Maria Emilia; Sosa, María de Los Ángeles; Fernández, Mariana; Rojas, Florencia; Mangiaterra, Magdalena; Giusiano, Gustavo

    2014-01-01

    In Argentina, information about epidemiology and environmental distribution of Cryptococcus is scarce. The city of Resistencia borders with Brazil and Paraguay where this fungus is endemic. All these supported the need to investigate the ecology of the genus and the epidemiology of cryptococcosis in this area. The aim was to investigate the presence of species of Cryptococcus neoformans-Cryptococcus gattii complex and their genotypes in trees of the city of Resistencia. One hundred and five trees were sampled by swabbing technique. The isolates were identified using conventional and commercial methods and genotyped by PCR-RFLP (Restriction Fragment Length Polymorphism). Cryptococcus was found in 7 out of the total trees. 6 out of 7 Cryptococcus isolates were identified as C. neoformans and one as C. gattii. C. gattii was isolated from Grevillea robusta. C. neoformans strains were isolated from Tabebuia avellanedae and Peltophorum dubium. Genotyping showed that all C. neoformans belonged to the VNI type and C. gattii belonged to the VGI type. This represents the first study on the ecology of Cryptococcus spp. associated to trees from northeastern Argentina, and the first report describing Grevillea robusta as a host of members of this fungal genus. Another finding is the isolation of C. neoformans from Tabebuia avellanedae and Peltophorum dubium, both tree species native to northeastern Argentina. Copyright © 2012 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  3. Continental Drift and Speciation of the Cryptococcus neoformans and Cryptococcus gattii Species Complexes

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    Arturo Casadevall

    2017-04-01

    Full Text Available Genomic analysis has placed the origins of two human-pathogenic fungi, the Cryptococcus gattii species complex and the Cryptococcus neoformans species complex, in South America and Africa, respectively. Molecular clock calculations suggest that the two species separated ~80 to 100 million years ago. This time closely approximates the breakup of the supercontinent Pangea, which gave rise to South America and Africa. On the basis of the geographic distribution of these two species complexes and the coincidence of the evolutionary divergence and Pangea breakup times, we propose that a spatial separation caused by continental drift resulted in the emergence of the C. gattii and C. neoformans species complexes from a Pangean ancestor. We note that, despite the spatial and temporal separation that occurred approximately 100 million years ago, these two species complexes are morphologically similar, share virulence factors, and cause very similar diseases. Continuation of these phenotypic characteristics despite ancient separation suggests the maintenance of similar selection pressures throughout geologic ages.

  4. Isolation of Cryptococcus gattii from a Castanopsis argyrophylla tree hollow (Mai-Kaw), Chiang Mai, Thailand.

    Science.gov (United States)

    Khayhan, Kantarawee; Hagen, Ferry; Norkaew, Treepradab; Puengchan, Tanpalang; Boekhout, Teun; Sriburee, Pojana

    2017-04-01

    The pathogenic yeast Cryptococcus gattii was isolated from a tree hollow of a Castanopsis argyrophylla King ex Hook.f. (Fagaceae) in Chiang Mai, Thailand. Molecular characterization with amplified fragment length polymorphism analysis and multi-locus sequence typing showed that this isolate belonged to genotype AFLP4/VGI representing C. gattii sensu stricto. Subsequent comparison of the environmental isolate with those from clinical samples from Thailand showed that they grouped closely together in a single cluster.

  5. Environmental distribution of Cryptococcus neoformans and C. gattii around the Mediterranean basin

    NARCIS (Netherlands)

    Cogliati, Massimo; D'Amicis, Roberta; Zani, Alberto; Montagna, Maria Teresa; Caggiano, Giuseppina; De Giglio, Osvalda; Balbino, Stella; De Donno, Antonella; Serio, Francesca; Susever, Serdar; Ergin, Cagri; Velegraki, Aristea; Ellabib, Mohamed S; Nardoni, Simona; Macci, Cristina; Oliveri, Salvatore; Trovato, Laura; Dipineto, Ludovico; Rickerts, Volker; McCormick-Smith, Ilka; Akcaglar, Sevim; Tore, Okan; Mlinaric-Missoni, Emilija; Bertout, Sebastien; Mallié, Michele; Martins, Maria da Luz; Vencà, Ana C F; Vieira, Maria L; Sampaio, Ana C; Pereira, Cheila; Griseo, Giuseppe; Romeo, Orazio; Ranque, Stéphane; Al-Yasiri, Mohammed H Y; Kaya, Meltem; Cerikcioglu, Nilgun; Marchese, Anna; Vezzulli, Luigi; Ilkit, Macit; Desnos-Ollivier, Marie; Pasquale, Vincenzo; Korem, Maya; Polacheck, Itzhack; Scopa, Antonio; Meyer, Wieland; Ferreira-Paim, Kennio; Hagen, Ferry; Theelen, Bart; Boekhout, Teun; Lockhart, Shawn R; Tintelnot, Kathrin; Tortorano, Anna Maria; Dromer, Françoise; Varma, Ashok; Kwon-Chung, Kyung J; Inácio, Joäo; Alonso, Beatriz; Colom, Maria F

    In order to elucidate the distribution of Cryptococcus neoformans and C. gattii in the Mediterranean basin, an extensive environmental survey was carried out during 2012-2015. A total of 302 sites located in 12 countries were sampled, 6436 samples from 3765 trees were collected and 5% of trees were

  6. Isolation of Cryptococcus gattii from a Castanopsis argyrophylla tree hollow (Mai-Kaw), Chiang Mai, Thailand

    NARCIS (Netherlands)

    Khayhan, Kantarawee; Hagen, Ferry; Norkaew, Treepradab; Puengchan, Tanpalang; Boekhout, Teun; Sriburee, Pojana

    2017-01-01

    The pathogenic yeast Cryptococcus gattii was isolated from a tree hollow of a Castanopsis argyrophylla King ex Hook.f. (Fagaceae) in Chiang Mai, Thailand. Molecular characterization with amplified fragment length polymorphism analysis and multi-locus sequence typing showed that this isolate belonged

  7. Genotypic and Phenotypic Diversity of Cryptococcus gattii VGII Clinical Isolates and Its Impact on Virulence

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    Vanessa A. Barcellos

    2018-02-01

    Full Text Available The Cryptococcus gattii species complex harbors the main etiological agents of cryptococcosis in immunocompetent patients. C. gattii molecular type VGII predominates in the north and northeastern regions of Brazil, leading to high morbidity and mortality rates. C. gattii VGII isolates have a strong clinical relevance and phenotypic variations. These phenotypic variations among C. gattii species complex isolates suggest that some strains are more virulent than others, but little information is available related to the pathogenic properties of those strains. In this study, we analyzed some virulence determinants of C. gattii VGII strains (CG01, CG02, and CG03 isolated from patients in the state of Piauí, Brazil. The C. gattii R265 VGIIa strain, which was isolated from the Vancouver outbreak, differed from C. gattii CG01, CG02 and CG03 isolates (also classified as VGII when analyzed the capsular dimensions, melanin production, urease activity, as well as the glucuronoxylomannan (GXM secretion. Those differences directly reflected in their virulence potential. In addition, CG02 displayed higher virulence compared to R265 (VGIIa strain in a cryptococcal murine model of infection. Lastly, we examined the genotypic diversity of these strains through Multilocus Sequence Type (MLST and one new subtype was described for the CG02 isolate. This study confirms the presence and the phenotypic and genotypic diversity of highly virulent strains in the Northeast region of Brazil.

  8. Clonality and α-a recombination in the Australian Cryptococcus gattii VGII population - an emerging outbreak in Australia

    NARCIS (Netherlands)

    Carriconde, Fabian; Gilgado, Félix; Arthur, Ian; Ellis, David; Malik, Richard; van de Wiele, Nathalie; Robert, Vincent; Currie, Bart J.; Meyer, Wieland

    2011-01-01

    Cryptococcus gattii is a basidiomycetous yeast that causes life-threatening disease in humans and animals. Within C. gattii, four molecular types are recognized (VGI to VGIV). The Australian VGII population has been in the spotlight since 2005, when it was suggested as the possible origin for the

  9. Ancient dispersal of the human fungal pathogen Cryptococcus gattii from the Amazon rainforest.

    Science.gov (United States)

    Hagen, Ferry; Ceresini, Paulo C; Polacheck, Itzhack; Ma, Hansong; van Nieuwerburgh, Filip; Gabaldón, Toni; Kagan, Sarah; Pursall, E Rhiannon; Hoogveld, Hans L; van Iersel, Leo J J; Klau, Gunnar W; Kelk, Steven M; Stougie, Leen; Bartlett, Karen H; Voelz, Kerstin; Pryszcz, Leszek P; Castañeda, Elizabeth; Lazera, Marcia; Meyer, Wieland; Deforce, Dieter; Meis, Jacques F; May, Robin C; Klaassen, Corné H W; Boekhout, Teun

    2013-01-01

    Over the past two decades, several fungal outbreaks have occurred, including the high-profile 'Vancouver Island' and 'Pacific Northwest' outbreaks, caused by Cryptococcus gattii, which has affected hundreds of otherwise healthy humans and animals. Over the same time period, C. gattii was the cause of several additional case clusters at localities outside of the tropical and subtropical climate zones where the species normally occurs. In every case, the causative agent belongs to a previously rare genotype of C. gattii called AFLP6/VGII, but the origin of the outbreak clades remains enigmatic. Here we used phylogenetic and recombination analyses, based on AFLP and multiple MLST datasets, and coalescence gene genealogy to demonstrate that these outbreaks have arisen from a highly-recombining C. gattii population in the native rainforest of Northern Brazil. Thus the modern virulent C. gattii AFLP6/VGII outbreak lineages derived from mating events in South America and then dispersed to temperate regions where they cause serious infections in humans and animals.

  10. Isolation of Cryptococcus gattii molecular type VGIII, from Corymbia ficifolia detritus in Colombia.

    Science.gov (United States)

    Escandón, P; Sánchez, A; Firacative, C; Castañeda, E

    2010-06-01

    An environmental sampling survey was carried out in different areas of Bogotá, Colombia, to obtain isolates of members of the Cryptococcus neoformans/C. gattii species complex from Corymbia ficifolia trees. During a 6-month period in 2007, 128 samples consisting of bark, soil around trunk bases, detritus, seeds and flowers were collected from 91 trees and processed according to standard procedures. The molecular type was determined using URA5 restriction fragment length polymorphism (RFLP) analysis and the mating type was established by PCR using specific primers for Mfalpha and Mfa C. gattii was isolated from 15 of the 128 (11.7%) samples, of which three (20%) were recovered from the red flower extract and the remaining 12 from C. ficifolia detritus. URA5 RFLP analysis revealed that all 15 isolates belonged to the molecular type VGIII and mating type specific PCR revealed that all were mating type a. The isolation of C. gattii from C. ficifolia represents an important finding since this is the first report revealing C. ficifolia as a habitat for C. gattii and adds additional information to the ever growing spectrum of tree species from which C. gattii can be recovered.

  11. First environmental isolation of Cryptococcus gattii serotype B, from Cúcuta, Colombia.

    Science.gov (United States)

    Firacative, Carolina; Torres, Germán; Rodríguez, María Claudia; Escandón, Patricia

    2011-03-01

    In Cúcuta, Cryptococcus gattii serotype B is commonly recovered from immunocompetent patients with cryptococcosis, but it has not been recovered from the environment in spite of its high incidence which is 77% out of reported cases. The aim of this work was to carry out an extensive environmental sampling in Cúcuta, in an attempt to isolate C. gattii serotype B and to expand our knowledge about the ecology and epidemiology of this important yeast. Samples associated with 3,634 trees from 40 zones of Cúcuta were collected and processed with 28 samples collected near the houses of four patients with cryptococcosis caused by C. gattii serotype B. The serotype of the recovered isolates was done using multiplex PCR, molecular patterns were determined by RFLP of the URA5 gene and mating type was determined using the primers MfαU, MfαL, MFa2U and MFa2L. In total, 4,389 samples were processed and one isolate of C. gattii serotype B (VGI/a), two isolates of C. gattii serotype C (VGIII/α) and three isolates of C. neoformans var. grubii, serotype A (VNI/α), were recovered. The density of the recovered isolates varied from 50 to 350 cfu/g of soil. This is the first report on the environmental isolation of C. gattii serotype B from Cúcuta. However, because of the low rate of recovery of isolates from soil only, the environmental niche of C. gattii has not been established and further environmental studies in Cúcuta are necessary, owing that this serotype is not only causing cryptococcosis but also has shown a higher virulence after the Vancouver outbreak.

  12. Transcriptional Analysis Allows Genome Reannotation and Reveals that Cryptococcus gattii VGII Undergoes Nutrient Restriction during Infection

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    Patrícia Aline Gröhs Ferrareze

    2017-08-01

    Full Text Available Cryptococcus gattii is a human and animal pathogen that infects healthy hosts and caused the Pacific Northwest outbreak of cryptococcosis. The inhalation of infectious propagules can lead to internalization of cryptococcal cells by alveolar macrophages, a niche in which C. gattii cells can survive and proliferate. Although the nutrient composition of macrophages is relatively unknown, the high induction of amino acid transporter genes inside the phagosome indicates a preference for amino acid uptake instead of synthesis. However, the presence of countable errors in the R265 genome annotation indicates significant inhibition of transcriptomic analysis in this hypervirulent strain. Thus, we analyzed RNA-Seq data from in vivo and in vitro cultures of C. gattii R265 to perform the reannotation of the genome. In addition, based on in vivo transcriptomic data, we identified highly expressed genes and pathways of amino acid metabolism that would enable C. gattii to survive and proliferate in vivo. Importantly, we identified high expression in three APC amino acid transporters as well as the GABA permease. The use of amino acids as carbon and nitrogen sources, releasing ammonium and generating carbohydrate metabolism intermediaries, also explains the high expression of components of several degradative pathways, since glucose starvation is an important host defense mechanism.

  13. Cryptococcus gattii: Emergence in Western North America: Exploitation of a Novel Ecological Niche

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    Kausik Datta

    2009-01-01

    Full Text Available The relatively uncommon fungal pathogen Cryptococcus gattii recently emerged as a significant cause of cryptococcal disease in human and animals in the Pacific Northwest of North America. Although genetic studies indicated its possible presence in the Pacific Northwest for more than 30 years, C. gattii as an etiological agent was largely unknown in this region prior to 1999. The recent emergence may have been encouraged by changing conditions of climate or land use and/or host susceptibility, and predictive ecological niche modeling indicates a potentially wider spread. C. gattii can survive wide climatic variations and colonize the environment in tropical, subtropical, temperate, and dry climates. Long-term climate changes, such as the significantly elevated global temperature in the last 100 years, influence patterns of disease among plants and animals and create niche microclimates habitable by emerging pathogens. C. gattii may have exploited such a hitherto unrecognized but clement environment in the Pacific Northwest to provide a wider exposure and risk of infection to human and animal populations.

  14. Polymorphism in Mitochondrial Group I Introns among Cryptococcus neoformans and Cryptococcus gattii Genotypes and Its Association with Drug Susceptibility

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    Felipe E. E. S. Gomes

    2018-02-01

    Full Text Available Cryptococcosis, one of the most important systemic mycosis in the world, is caused by different genotypes of Cryptococcus neoformans and Cryptococcus gattii, which differ in their ecology, epidemiology, and antifungal susceptibility. Therefore, the search for new molecular markers for genotyping, pathogenicity and drug susceptibility is necessary. Group I introns fulfill the requisites for such task because (i they are polymorphic sequences; (ii their self-splicing is inhibited by some drugs; and (iii their correct splicing under parasitic conditions is indispensable for pathogen survival. Here, we investigated the presence of group I introns in the mitochondrial LSU rRNA gene in 77 Cryptococcus isolates and its possible relation to drug susceptibility. Sequencing revealed two new introns in the LSU rRNA gene. All the introns showed high sequence similarity to other mitochondrial introns from distinct fungi, supporting the hypothesis of an ancient non-allelic invasion. Intron presence was statistically associated with those genotypes reported to be less pathogenic (p < 0.001. Further virulence assays are needed to confirm this finding. In addition, in vitro antifungal tests indicated that the presence of LSU rRNA introns may influence the minimum inhibitory concentration (MIC of amphotericin B and 5-fluorocytosine. These findings point to group I introns in the mitochondrial genome of Cryptococcus as potential molecular markers for antifungal resistance, as well as therapeutic targets.

  15. Cryptococcus neoformans and Cryptococcus gattii isolated from the excreta of psittaciformes in a southern Brazilian zoological garden.

    Science.gov (United States)

    Abegg, Maxwel Adriano; Cella, Fabiana Lucila; Faganello, Josiane; Valente, Patrícia; Schrank, Augusto; Vainstein, Marilene Henning

    2006-02-01

    Cryptococcus neoformans, a major pathogen in immunocompromised patients, is a ubiquitous free-living fungus that can be isolated from soils, avian excreta and plant material. To further study potential saprophytic sources of this yeast in the Southern Brazilian State Rio Grande do Sul, we analyzed fecal samples from 59 species of captive birds kept in cages at a local Zoological Garden, belonging to 12 different orders. Thirty-eight environmental isolates of C. neoformans were obtained only from Psittaciformes (Psittacidae, Cacatuidae and Psittacula). Their variety and serotype were determined, and the genetic structure of the isolates was analyzed by use of the simple repetitive microsatellite specific primer M13 and the minisatellite specific primer (GACA)(4) as single primers in the PCR. The varieties were confirmed by pulsed-field gel electrophoresis (PFGE). Thirty-three isolates (87%) were from the var. grubii, serotype A, molecular type VNI and five (13%) were Cryptococcus gattii, serotype B, molecular type VGI. All the isolates were mating type alpha. Isolates were screened for some potential virulence factors. Quantitative urease production by the environmental isolates belonging to the C. gattii was similar to the values usually obtained for clinical ones.

  16. Clinical and microbiological characteristics of cryptococcosis in Singapore: predominance of Cryptococcus neoformans compared with Cryptococcus gattii

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    Monica Chan

    2014-09-01

    Conclusion: C. neoformans var. grubii, subtype VN I, was the predominant subtype in Singapore, infecting younger, mainly immunocompromised hosts with HIV. C. gattii was uncommon, causing pulmonary manifestations in older, immunocompetent patients and were RFLP type VG II.

  17. Modulation of Zinc Homeostasis in Acanthamoeba castellanii as a Possible Antifungal Strategy against Cryptococcus gattii

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    Nicole S. Ribeiro

    2017-08-01

    Full Text Available Cryptococcus gattii is a basidiomycetous yeast that can be found in the environment and is one of the agents of cryptococcosis, a life-threatening disease. During its life cycle, cryptococcal cells take hold inside environmental predators such as amoebae. Despite their evolutionary distance, macrophages and amoebae share conserved similar steps of phagocytosis and microbial killing. To evaluate whether amoebae also share other antifungal strategies developed by macrophages, we investigated nutritional immunity against cryptococcal cells. We focused on zinc homeostasis modulation in Acanthamoeba castellanii infected with C. gattii. The intracellular proliferation rate (IPR in amoebae was determined using C. gattii R265 and mutants for the ZIP1 gene, which displays defects of growth in zinc-limiting conditions. We detected a reduced IPR in cells lacking the ZIP1 gene compared to wild-type strains, suggesting that amoebae produce a low zinc environment to engulfed cells. Furthermore, flow cytometry analysis employing the zinc probe Zinpyr-1 confirmed the reduced concentration of zinc in cryptococcal-infected amoebae. qRT-PCR analysis of zinc transporter-coding genes suggests that zinc export by members of the ZnT family would be involved in the reduced intracellular zinc concentration. These results indicate that amoebae may use nutritional immunity to reduce fungal cell proliferation by reducing zinc availability for the pathogen.

  18. Preshipment testing success: resolution of a nasal sinus granuloma in a captive koala (Phascolarctos cinereus) caused by Cryptococcus gattii.

    Science.gov (United States)

    Wynne, Janna; Klause, Stephen; Stadler, Cynthia K; Pye, Geoffrey W; Meyer, Wieland; Sykes, Jane E

    2012-12-01

    A 3-yr-old female koala (Phascolarctos cinereus) was diagnosed with a nasal sinus granuloma caused by Cryptococcus gattii after a pre-shipment examination revealed a latex cryptococcal agglutination titer of 1:512. Successful medical and surgical treatment of the granuloma was monitored using serial latex cryptococcal agglutination titers, serum levels of antifungal drugs, and advanced imaging.

  19. Galleria mellonella model identifies highly virulent strains among all major molecular types of Cryptococcus gattii.

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    Carolina Firacative

    Full Text Available Cryptococcosis is mainly caused by Cryptococcus neoformans. However, the number of cases due to C. gattii is increasing, affecting mainly immunocompetent hosts. C. gattii is divided into four major molecular types, VGI to VGIV, which differ in their host range, epidemiology, antifungal susceptibility and geographic distribution. Besides studies on the Vancouver Island outbreak strains, which showed that the subtype VGIIa is highly virulent compared to the subtype VGIIb, little is known about the virulence of the other major molecular types. To elucidate the virulence potential of the major molecular types of C. gattii, Galleria mellonella larvae were inoculated with ten globally selected strains per molecular type. Survival rates were recorded and known virulence factors were studied. One VGII, one VGIII and one VGIV strain were more virulent (p 0.05, 21 (five VGI, five VGII, four VGIII and seven VGIV were less virulent (p <0.05 while one strain of each molecular type were avirulent. Cell and capsule size of all strains increased markedly during larvae infection (p <0.001. No differences in growth rate at 37°C were observed. Melanin synthesis was directly related with the level of virulence: more virulent strains produced more melanin than less virulent strains (p <0.05. The results indicate that all C. gattii major molecular types exhibit a range of virulence, with some strains having the potential to be more virulent. The study highlights the necessity to further investigate the genetic background of more and less virulent strains in order to recognize critical features, other than the known virulence factors (capsule, melanin and growth at mammalian body temperature, that maybe crucial for the development and progression of cryptococcosis.

  20. Impact of Resistance to Fluconazole on Virulence and Morphological Aspects of Cryptococcus neoformans and Cryptococcus gattii Isolates.

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    Suélen Andreia eRossi

    2016-02-01

    Full Text Available Cryptococcus spp. are responsible for around one million cases of meningitis every year. Fluconazole (FLU is commonly used in the treatment of cryptococcosis, mainly in immunocompromised patients and the resistance is usually reported after long periods of treatment. In this study, the morphological characterization and virulence profile of FLU-susceptible and FLU-resistant clinical and environmental isolates of C. neoformans and C. gattii were performed both in vitro and in vivo using the Galleria mellonella model. FLU-susceptible isolates from C. neoformans were significantly more virulent than the FLU-resistant isolates. FLU-susceptible C. gattii isolates showed a different virulence profile from C. neoformans isolates where only the environmental isolate, CL, was more virulent compared with the resistant isolates. Cell morphology and capsule size were analyzed and the FLU-resistant isolates did not change significantly compared with the most sensitive isolates. Growth at 37°C was also evaluated and in both species, the resistant isolates showed a reduced growth at this temperature, indicating that FLU resistance can affect their growth. Based on the results obtained is possible suggest that FLU resistance can influence the morphology of the isolates and consequently changed the virulence profiles. The most evident results were observed for C. neoformans showing that the adaptation of isolates to antifungal selective pressure influenced the loss of virulence.

  1. Cryptococcus neoformans in the koala (Phascolarctos cinereus): colonization by C n. var. gattii and investigation of environmental sources.

    Science.gov (United States)

    Krockenberger, M B; Canfield, P J; Malik, R

    2002-06-01

    This study is the one in a series looking at the relationship among Cryptococcus neoformans var. gattii, koalas and the environment. The koala was used as a natural biological sampler in an attempt to understand the dynamics of C. neoformans var. gattii in Australian environments. Evidence of asymptomatic nasal and skin colonization for extended periods by large numbers of C. n. var. gattii was obtained and geographical factors assessed. The key finding was the ability of koalas to amplify numbers of C. n. var. gattii in certain environments. Koalas were not found to be obligatory for the survival of the organism in all environments. Geographical factors alone could not explain differing rates of nasal and skin colonization in koalas in different environments. A strong association between healthy koalas and C. n. var. gattii was confirmed and C n. var. gattii was isolated from novel sources, including the turpentine gum tree (Syncarpia glomulifera), tallowwood (Eucalyptus microcorys) and flooded gum (E. grandis). It seems likely that as yet undiscovered environmental sources of C. n. var. gattii exist in eastern Australia. Further investigation of host, environmental and organism factors integral to the hostpathogen relationship will assist an understanding of the progression from colonization to tissue invasion and cryptococcosis in all species.

  2. [Presumptive identification of Cryptococcus gattii isolated from Terminalia catappa in Montería city, Córdoba, Colombia].

    Science.gov (United States)

    Contreras Martínez, Orfa Inés; Aycardi Morinelli, María Paulina; Alarcón Furnieles, Jany Luz; Jaraba Ramos, Aparicio Manuel

    2011-01-01

    the members of the Cryptococcus neoformans species complex are responsible for cryptococcosis in animals and humans. Human infection is thought to be acquired by inhalation of airborne propagules from an environmental source; therefore it is greatly important to study their habitat. to determine the ecological relationship of Cryptococcus gattii with Terminalia catappa trees present in urban areas of Montería city in Colombia. a total of 163 Terminalia catappa trees were selected; some samples were taken from the bark, the leaves, the flowers, the fruits of these trees and from the surrounding soil. The yeast was isolated using the Guizotia abyssinica seed agar medium; it was identified thanks to biochemical and morphologic tests whereas the right variety was determined by L-canavanine-glycine-bromothymol blue (CGB), D-proline and D-tryptophan tests. there was obtained 9.050 CFU/g isolate of Cryptococcus spp., 5.795 CFU/g of which were presumptively identified as Cryptococcus gattii. The highest percentage of isolates was found in flowers, followed by bark and fruits, presenting small cellular and capsular sizes. These isolates were more frequent in the south of the city, followed by the center zone and the lowest percentage in the northern zone. these findings confirmed the close relationship of Cryptococcus gattii and Terminalia catappa, being this the first study conducted in Monteria city. These results give us meaningful information for understanding and analyzing the epidemiology of cryptococcosis in Monteria city, Colombia.

  3. Antifungal activity of eicosanoic acids isolated from the endophytic fungus Mycosphaerella sp. against Cryptococcus neoformans and C. gattii.

    Science.gov (United States)

    Pereira, Cristiane Bigatti; Pereira de Sá, Nívea; Borelli, Beatriz Martins; Rosa, Carlos Augusto; Barbeira, Paulo Jorge Sanches; Cota, Betania Barros; Johann, Susana

    2016-11-01

    The antifungal effects of two eicosanoic acids, 2-amino-3,4-dihydroxy-2-25-(hydroxymethyl)-14-oxo-6,12-eicosenoic acid (compound 1) and myriocin (compound 2), isolated from Mycosphaerella sp. were evaluated against Cryptococcus neoformans and C. gattii. The compounds displayed antifungal activities against several isolates of C. neoformans and C. gattii, with minimal inhibitory concentration (MIC) values ranging from 0.49 to 7.82 μM for compound 1 and 0.48-1.95 μM for compound 2. In the checkerboard microtiter test, both compounds exhibited synergistic activity with amphotericin B against C. gattii. Ultrastructural analysis revealed several signs of damage in C. gattii and C. neoformans cells treated with compounds 1 and 2, including deformities in cell shape, depressions on the surface, and withered cells. The cells of C. gattii treated with compounds 1 and 2 showed less loss of cellular material in comparison to those treated with amphotericin B. The difference in cellular material loss increased in a test compound concentration-dependent manner. Consistent with this observation, compounds 1 and 2 were able to internalize propidium iodide (PI) in C. gattii cells. In addition, compound 2 induced the formation of several pseudohyphae, suggesting that it could reduce virulence in C. gattii cells. The study results show that these natural products led to membrane damage; however, this may not be the main target of action. These compounds have potential antifungal activity and could be useful in further studies for developing more effective combination therapies with amphotericin B and reducing side effects in patients. Copyright © 2016 Elsevier Ltd. All rights reserved.

  4. Cryptococcus gattii urease as a virulence factor and the relevance of enzymatic activity in cryptococcosis pathogenesis.

    Science.gov (United States)

    Feder, Vanessa; Kmetzsch, Lívia; Staats, Charley Christian; Vidal-Figueiredo, Natalia; Ligabue-Braun, Rodrigo; Carlini, Célia Regina; Vainstein, Marilene Henning

    2015-04-01

    Ureases (EC 3.5.1.5) are Ni(2+) -dependent metalloenzymes produced by plants, fungi and bacteria that hydrolyze urea to produce ammonia and CO2 . The insertion of nickel atoms into the apo-urease is better characterized in bacteria, and requires at least three accessory proteins: UreD, UreF, and UreG. Our group has demonstrated that ureases possess ureolytic activity-independent biological properties that could contribute to the pathogenicity of urease-producing microorganisms. The presence of urease in pathogenic bacteria strongly correlates with pathogenesis in some human diseases. Some medically important fungi also produce urease, including Cryptococcus neoformans and Cryptococcus gattii. C. gattii is an etiological agent of cryptococcosis, most often affecting immunocompetent individuals. The cryptococcal urease might play an important role in pathogenesis. It has been proposed that ammonia produced via urease action might damage the host endothelium, which would enable yeast transmigration towards the central nervous system. To analyze the role of urease as a virulence factor in C. gattii, we constructed knockout mutants for the structural urease-coding gene URE1 and for genes that code the accessory proteins Ure4 and Ure6. All knockout mutants showed reduced multiplication within macrophages. In intranasally infected mice, the ure1Δ (lacking urease protein) and ure4Δ (enzymatically inactive apo-urease) mutants caused reduced blood burdens and a delayed time of death, whereas the ure6Δ (enzymatically inactive apo-urease) mutant showed time and dose dependency with regard to fungal burden. Our results suggest that C. gattii urease plays an important role in virulence, in part possibly through enzyme activity-independent mechanism(s). © 2015 FEBS.

  5. Intestinal Lesion in a Dog Due to Cryptococcus gattii Type VGII and Review of Published Cases of Canine Gastrointestinal Cryptococcosis.

    Science.gov (United States)

    de Abreu, Daniel Paiva Barros; Machado, Carlos Henrique; Makita, Mário Tatsuo; Botelho, Camila Flávia Magalhães; Oliveira, Fernanda Gomes; da Veiga, Cristiano Chaves Pessoa; Martins, Marilena Dos Anjos; Baroni, Francisco de Assis

    2017-06-01

    Cryptococcosis is a mycosis caused by yeasts of genus Cryptococcus, mainly the species C. neoformans and C. gattii that can affect humans and animals. These yeasts are widely distributed in the environment and are typically associated with avian droppings and decaying wood. Most infections are related to the respiratory tract, but the central nervous system and cutaneous lesions are also reported in the literature. The present report is a case of cryptococcosis in an 18-month-old unspayed female English Bulldog with the main complaint of weight loss and diarrhea. The presence of two large masses observed in an ultrasound examination leads us to perform an exploratory laparotomy. Considering the size of the lesion and the impossibility of owner to provide intensive care, the consent for euthanasia was requested. The postmortem diagnosis of cryptococcosis was revealed by cytological evaluation, and the involvement of C. gattii VGII was confirmed by isolation and identification tests as well as by the detection of the URA5 gene restriction fragment length polymorphism PCR analysis. Reports in the literature of the involvement of Cryptococcus in gastrointestinal lesions are rare in both human and veterinary medicine. Data about different forms of cryptococcosis are important to provide more knowledge of uncommon clinical presentations of this yeast and therefore improve the diagnoses and decisions for the best therapy.

  6. Clonality and α-a recombination in the Australian Cryptococcus gattii VGII population--an emerging outbreak in Australia.

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    Fabian Carriconde

    Full Text Available BACKGROUND: Cryptococcus gattii is a basidiomycetous yeast that causes life-threatening disease in humans and animals. Within C. gattii, four molecular types are recognized (VGI to VGIV. The Australian VGII population has been in the spotlight since 2005, when it was suggested as the possible origin for the ongoing outbreak at Vancouver Island (British Columbia, Canada, with same-sex mating being suggested as the driving force behind the emergence of this outbreak, and is nowadays hypothesized as a widespread phenomenon in C. gattii. However, an in-depth characterization of the Australian VGII population is still lacking. The present work aimed to define the genetic variability within the Australian VGII population and determine processes shaping its population structure. METHODOLOGY/PRINCIPAL FINDINGS: A total of 54 clinical, veterinary and environmental VGII isolates from different parts of the Australian continent were studied. To place the Australian population in a global context, 17 isolates from North America, Europe, Asia and South America were included. Genetic variability was assessed using the newly adopted international consensus multi-locus sequence typing (MLST scheme, including seven genetic loci: CAP59, GPD1, LAC1, PLB1, SOD1, URA5 and IGS1. Despite the overall clonality observed, the presence of MATa VGII isolates in Australia was demonstrated for the first time in association with recombination in MATα-MATa populations. Our results also support the hypothesis of a "smouldering" outbreak throughout the Australian continent, involving a limited number of VGII genotypes, which is possibly caused by a founder effect followed by a clonal expansion. CONCLUSIONS/SIGNIFICANCE: The detection of sexual recombination in MATα-MATa population in Australia is in accordance with the natural life cycle of C. gattii involving opposite mating types and presents an alternative to the same-sex mating strategy suggested elsewhere. The potential

  7. First reported case of Cryptococcus gattii in the Southeastern USA: implications for travel-associated acquisition of an emerging pathogen.

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    Edmond J Byrnes

    2009-06-01

    Full Text Available In 2007, the first confirmed case of Cryptococcus gattii was reported in the state of North Carolina, USA. An otherwise healthy HIV negative male patient presented with a large upper thigh cryptococcoma in February, which was surgically removed and the patient was started on long-term high-dose fluconazole treatment. In May of 2007, the patient presented to the Duke University hospital emergency room with seizures. Magnetic resonance imaging revealed two large CNS lesions found to be cryptococcomas based on brain biopsy. Prior chest CT imaging had revealed small lung nodules indicating that C. gattii spores or desiccated yeast were likely inhaled into the lungs and dissemination occurred to both the leg and CNS. The patient's travel history included a visit throughout the San Francisco, CA region in September through October of 2006, consistent with acquisition during this time period. Cultures from both the leg and brain biopsies were subjected to analysis. Based on phenotypic and molecular methods, both isolates were C. gattii, VGI molecular type, and distinct from the Vancouver Island outbreak isolates. Based on multilocus sequence typing of coding and noncoding regions and virulence in a heterologous host model, the leg and brain isolates are identical, but the two differed in mating fertility. Two clinical isolates, one from a transplant recipient in San Francisco and the other from Australia, were identical to the North Carolina clinical isolate at all markers tested. Closely related isolates that differ at only one or a few noncoding markers are present in the Australian environment. Taken together, these findings support a model in which C. gattii VGI was transferred from Australia to California, possibly though an association with its common host plant E. camaldulensis, and the patient was exposed in San Francisco and returned to present with disease in North Carolina.

  8. Environmental sampling of Ceratonia siliqua (carob) trees in Spain reveals the presence of the rare Cryptococcus gattii genotype AFLP7/VGIV.

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    Linares, Carlos; Colom, María Francisca; Torreblanca, Marina; Esteban, Violeta; Romera, Álvaro; Hagen, Ferry

    2015-01-01

    Cryptococcus gattii is a pathogenic basidiomycetous yeast that is emerging in temperate climate zones worldwide. C. gattii has repetitively been isolated from numerous tree species. Ongoing environmental sampling and molecular characterization is essential to understand the presence of this primary pathogenic microorganism in the Mediterranean environment. To report the first isolation of the rare C. gattii genotype AFLP7/VGIV from the environment in Europe. Samples were collected from woody debris of carob trees (Ceratonia siliqua) and olive trees (Olea europaea) in El Perelló, Tarragona, Spain. Cryptococcus species were further characterized by using URA5-RFLP, MALDI-TOF, AFLP and MLST. The antifungal susceptibility profile to amphotericin B, 5-fluorocytosine, fluconazole, itraconazole, posaconazole and voriconazole was determined using Sensititre Yeast One and E-test. Cultures from one carob tree revealed the presence of ten Cryptococcus-like colonies. One colony was identified as C. gattii, and subsequent molecular characterization showed that it was an α mating-type that belonged to the rare genotype AFLP7/VGIV. Antifungal susceptibility testing showed values within the range of sensitivity described for other isolates of the same genotype and within the epidemiological cutoff values for this species. The isolation of the rare C. gattii genotype AFLP7/VGIV in Spain is the first report in the European environment, implying the possible presence in other regions of the Mediterranean area, and underlines that clinicians must be aware for C. gattii infections in healthy individuals. Copyright © 2014 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  9. Cryptococcus gattii in an Immunocompetent Patient in the Southeastern United States

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    John W. Amburgy

    2016-01-01

    Full Text Available Cryptococcal infections are seen throughout the United States in both immunocompromised and immunocompetent patients. The most common form is C. neoformans. In the Northwestern United States, C. gattii has received considerable attention secondary to increased virulence resulting in significant morbidity and mortality. There are no cases in the extant literature describing a patient with C. gattii requiring neurosurgical intervention in Alabama. A middle-aged immunocompetent male with no recent travel or identifiable exposure presented with meningitis secondary to C. gattii. The patient underwent 12 lumbar punctures and a ventriculoperitoneal shunt and required 83 days of inpatient therapy with 5-flucytosine and amphotericin B. The patient was found to have multiple intracranial lesions and a large intramedullary spinal cryptococcoma within his conus. Following an almost 3-month hospitalization the patient required treatment with oral voriconazole for one year. In the United States meningitis caused by C. gattii infection is not isolated to the Northwestern region.

  10. Carbon dioxide is a powerful inducer of monokaryotic hyphae and spore development in Cryptococcus gattii and carbonic anhydrase activity is dispensable in this dimorphic transition.

    Science.gov (United States)

    Ren, Ping; Chaturvedi, Vishnu; Chaturvedi, Sudha

    2014-01-01

    Cryptococcus gattii is unique among human pathogenic fungi with specialized ecological niche on trees. Since leaves concentrate CO2, we investigated the role of this gaseous molecule in C. gattii biology and virulence. We focused on the genetic analyses of β-carbonic anhydrase (β-CA) encoded by C. gattii CAN1 and CAN2 as later is critical for CO2 sensing in a closely related pathogen C. neoformans. High CO2 conditions induced robust development of monokaryotic hyphae and spores in C. gattii. Conversely, high CO2 completely repressed hyphae development in sexual mating. Both CAN1 and CAN2 were dispensable for CO2 induced morphogenetic transitions. However, C. gattii CAN2 was essential for growth in ambient air similar to its reported role in C. neoformans. Both can1 and can2 mutants retained full pathogenic potential in vitro and in vivo. These results provide insight into C. gattii adaptation for arboreal growth and production of infectious propagules by β-CA independent mechanism(s).

  11. Genotipificación de aislamientos clínicos del complejo Cryptococcus neoformans/Cryptococcus gattii obtenidos en el Hospital «Dr. Julio C. Perrando», de la ciudad de Resistencia (Chaco, Argentina

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    Maria E Cattana

    Full Text Available La criptococosis es una infección fúngica causada por levaduras del género Cryptococcus, particularmente las del complejo Cryptococcus neoformans/Cryptococcus gattii. El conocimiento sobre la casuística de la criptococosis en el nordeste argentino es exiguo y no se tiene información sobre los tipos moleculares circulantes. El objetivo de este estudio fue realizar la caracterización genética de los aislamientos pertenecientes al complejo C. neoformans/C. gattii obtenidos en el Hospital «Dr. Julio C. Perrando» de la ciudad de Resistencia (Chaco, Argentina, con el fin de determinar especie, variedad y genotipo. Durante dos años y un mes se estudiaron 26 aislamientos clínicos. Mediante métodos convencionales y moleculares, un aislamiento fue identificado como C. gattii genotipo VGI y los 25 restantes como C. neoformans var.grubii, 23 de los cuales correspondieron al genotipo VNI y dos al genotipo VNII. Estos datos son una contribución al conocimiento de la epidemiología de la criptococosis en la Argentina y el primer informe sobre genotipos del complejo C. neoformans/C. gattii de origen clínico en el nordeste argentino.

  12. Cryptococcus gattii VGIII isolates causing infections in HIV/AIDS patients in Southern California: identification of the local environmental source as arboreal.

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    Deborah J Springer

    2014-08-01

    Full Text Available Ongoing Cryptococcus gattii outbreaks in the Western United States and Canada illustrate the impact of environmental reservoirs and both clonal and recombining propagation in driving emergence and expansion of microbial pathogens. C. gattii comprises four distinct molecular types: VGI, VGII, VGIII, and VGIV, with no evidence of nuclear genetic exchange, indicating these represent distinct species. C. gattii VGII isolates are causing the Pacific Northwest outbreak, whereas VGIII isolates frequently infect HIV/AIDS patients in Southern California. VGI, VGII, and VGIII have been isolated from patients and animals in the Western US, suggesting these molecular types occur in the environment. However, only two environmental isolates of C. gattii have ever been reported from California: CBS7750 (VGII and WM161 (VGIII. The incongruence of frequent clinical presence and uncommon environmental isolation suggests an unknown C. gattii reservoir in California. Here we report frequent isolation of C. gattii VGIII MATα and MATa isolates and infrequent isolation of VGI MATα from environmental sources in Southern California. VGIII isolates were obtained from soil debris associated with tree species not previously reported as hosts from sites near residences of infected patients. These isolates are fertile under laboratory conditions, produce abundant spores, and are part of both locally and more distantly recombining populations. MLST and whole genome sequence analysis provide compelling evidence that these environmental isolates are the source of human infections. Isolates displayed wide-ranging virulence in macrophage and animal models. When clinical and environmental isolates with indistinguishable MLST profiles were compared, environmental isolates were less virulent. Taken together, our studies reveal an environmental source and risk of C. gattii to HIV/AIDS patients with implications for the >1,000,000 cryptococcal infections occurring annually for which

  13. MALDI-TOF MS enables the rapid identification of the major molecular types within the Cryptococcus neoformans/C. gattii species complex.

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    Carolina Firacative

    Full Text Available BACKGROUND: The Cryptococcus neoformans/C. gattii species complex comprises two sibling species that are divided into eight major molecular types, C. neoformans VNI to VNIV and C. gattii VGI to VGIV. These genotypes differ in host range, epidemiology, virulence, antifungal susceptibility and geographic distribution. The currently used phenotypic and molecular identification methods for the species/molecular types are time consuming and expensive. As Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry (MALDI-TOF MS offers an effective alternative for the rapid identification of microorganisms, the objective of this study was to examine its potential for the identification of C. neoformans and C. gattii strains at the intra- and inter-species level. METHODOLOGY: Protein extracts obtained via the formic acid extraction method of 164 C. neoformans/C. gattii isolates, including four inter-species hybrids, were studied. RESULTS: The obtained mass spectra correctly identified 100% of all studied isolates, grouped each isolate according to the currently recognized species, C. neoformans and C. gattii, and detected potential hybrids. In addition, all isolates were clearly separated according to their major molecular type, generating greater spectral differences among the C. neoformans molecular types than the C. gattii molecular types, most likely reflecting a closer phylogenetic relationship between the latter. The number of colonies used and the incubation length did not affect the results. No spectra were obtained from intact yeast cells. An extended validated spectral library containing spectra of all eight major molecular types was established. CONCLUSIONS: MALDI-TOF MS is a rapid identification tool for the correct recognition of the two currently recognized human pathogenic Cryptococcus species and offers a simple method for the separation of the eight major molecular types and the detection of hybrid strains within this

  14. Retrospective Study of the Epidemiology and Clinical Manifestations of Cryptococcus gattii Infections in Colombia from 1997–2011

    Science.gov (United States)

    Lizarazo, Jairo; Escandón, Patricia; Agudelo, Clara Inés; Firacative, Carolina; Meyer, Wieland; Castañeda, Elizabeth

    2014-01-01

    Background Cryptococcosis due to Cryptococcus gattii is endemic in various parts of the world, affecting mostly immunocompetent patients. A national surveillance study of cryptococcosis, including demographical, clinical and microbiological data, has been ongoing since 1997 in Colombia, to provide insights into the epidemiology of this mycosis. Methodology/Principal Findings From 1,209 surveys analyzed between 1997–2011, 45 cases caused by C. gattii were reported (prevalence 3.7%; annual incidence 0.07 cases/million inhabitants/year). Norte de Santander had the highest incidence (0.81 cases/million/year), representing 33.3% of all cases. The male: female ratio was 3.3∶1. Mean age at diagnosis was 41±16 years. No specific risk factors were identified in 91.1% of patients. HIV infection was reported in 6.7% of patients, autoimmune disease and steroids use in 2.2%. Clinical features included headache (80.5%), nausea/vomiting (56.1%) and neurological derangements (48.8%). Chest radiographs were taken in 21 (46.7%) cases, with abnormal findings in 7 (33.3%). Cranial CT scans were obtained in 15 (33.3%) cases, with abnormalities detected in 10 (66.7%). Treatment was well documented in 30 cases, with most receiving amphotericin B. Direct sample examination was positive in 97.7% cases. Antigen detection was positive for all CSF specimens and for 75% of serum samples. C. gattii was recovered from CSF (93.3%) and respiratory specimens (6.6%). Serotype was determined in 42 isolates; 36 isolates were serotype B (85.7%), while 6 were C (14.3%). The breakdowns of molecular types were VGII (55.6%), VGIII (31.1%) and VGI (13.3%). Among 44 strains, 16 MLST sequence types (ST) were identified, 11 of them newly reported. Conclusions/Significance The results of this passive surveillance study demonstrate that cryptococcosis caused by C. gattii has a low prevalence in Colombia, with the exception of Norte de Santander. The predominance of molecular type VGII is of concern

  15. Population Genetic Analysis Reveals a High Genetic Diversity in the Brazilian Cryptococcus gattii VGII Population and Shifts the Global Origin from the Amazon Rainforest to the Semi-arid Desert in the Northeast of Brazil.

    Science.gov (United States)

    Souto, Ana C P; Bonfietti, Lucas X; Ferreira-Paim, Kennio; Trilles, Luciana; Martins, Marilena; Ribeiro-Alves, Marcelo; Pham, Cau D; Martins, Liline; Dos Santos, Wallace; Chang, Marilene; Brito-Santos, Fabio; Santos, Dayane C S; Fortes, Silvana; Lockhart, Shawn R; Wanke, Bodo; Melhem, Márcia S C; Lazéra, Márcia S; Meyer, Wieland

    2016-08-01

    Cryptococcus neoformans and Cryptococcus gattii are responsible globally for almost one million cryptococcosis cases yearly, mostly in immunocompromised patients, such as those living with HIV. Infections due to C. gattii have mainly been described in tropical and subtropical regions, but its adaptation to temperate regions was crucial in the species evolution and highlighted the importance of this pathogenic yeast in the context of disease. Cryptococcus gattii molecular type VGII has come to the forefront in connection with an on-going emergence in the Pacific North West of North America. Taking into account that previous work pointed towards South America as an origin of this species, the present work aimed to assess the genetic diversity within the Brazilian C. gattii VGII population in order to gain new insights into its origin and global dispersal from the South American continent using the ISHAM consensus MLST typing scheme. Our results corroborate the finding that the Brazilian C. gattii VGII population is highly diverse. The diversity is likely due to recombination generated from sexual reproduction, as evidenced by the presence of both mating types in clinical and environmental samples. The data presented herein strongly supports the emergence of highly virulent strains from ancestors in the Northern regions of Brazil, Amazonia and the Northeast. Numerous genotypes represent a link between Brazil and other parts of the world reinforcing South America as the most likely origin of the C. gattii VGII subtypes and their subsequent global spread, including their dispersal into North America, where they caused a major emergence.

  16. Endogenous endophthalmitis caused by Cryptococcus neoformans Var. Gattii mimicking choroidal tumor: From positron-emission tomography/computed tomography to histopathology

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    Chun-Ju Lin

    2017-01-01

    Full Text Available A 60-year-old immunocompetent female with pneumonia history about 10 years ago suffered from blurred vision overall survival for 3 weeks. Common cold with headache and unintentional body weight loss was also noted recently. Choroidal detachment simulating choroidal tumor was observed in the temporal quadrant. The 18F-fluoro-2-deoxyglucose (FDG positron-emission tomography/computed tomography (CT scan showed focal, mild to moderate FDG uptake in the left lateral orbit, and an enhanced lesion was seen on the recent CT scan. The suspicious choroidal tumor became larger in a week. Phacoemulsification, vitrectomy, and retinal biopsies were performed. Histochemical study of the retinal specimens established the diagnosis of endogenous cryptococcal endophthalmitis. Vitreous culture yielded Cryptococcus neoformans var. gattii. Systemic and topical voriconazole eliminated the infection. In the literature, endogenous endophthalmitis caused by C. neoformans var. gattii has not been reported in detail. The key to successful management lies in early diagnosis. If clinical improvement could not be achieved after conventional management and imaging studies have failed to yield a definite diagnosis, retinal biopsy can be considered.

  17. Molecular Epidemiology Reveals Genetic Diversity amongst Isolates of the Cryptococcus neoformans/C. gattii Species Complex in Thailand

    Science.gov (United States)

    Kaocharoen, Sirada; Ngamskulrungroj, Popchai; Firacative, Carolina; Trilles, Luciana; Piyabongkarn, Dumrongdej; Banlunara, Wijit; Poonwan, Natteewan; Chaiprasert, Angkana; Meyer, Wieland; Chindamporn, Ariya

    2013-01-01

    To gain a more detailed picture of cryptococcosis in Thailand, a retrospective study of 498 C. neoformans and C. gattii isolates has been conducted. Among these, 386, 83 and 29 strains were from clinical, environmental and veterinary sources, respectively. A total of 485 C. neoformans and 13 C. gattii strains were studied. The majority of the strains (68.9%) were isolated from males (mean age of 37.97 years), 88.5% of C. neoformans and only 37.5% of C. gattii strains were from HIV patients. URA5-RFLP and/or M13 PCR-fingerprinting analysis revealed that the majority of the isolates were C. neoformans molecular type VNI regardless of their sources (94.8%; 94.6% of the clinical, 98.8% of the environmental and 86.2% of the veterinary isolates). In addition, the molecular types VNII (2.4%; 66.7% of the clinical and 33.3% of the veterinary isolates), VNIV (0.2%; 100% environmental isolate), VGI (0.2%; 100% clinical isolate) and VGII (2.4%; 100% clinical isolates) were found less frequently. Multilocus Sequence Type (MLST) analysis using the ISHAM consensus MLST scheme for the C. neoformans/C. gattii species complex identified a total of 20 sequence types (ST) in Thailand combining current and previous data. The Thai isolates are an integrated part of the global cryptococcal population genetic structure, with ST30 for C. gattii and ST82, ST83, ST137, ST141, ST172 and ST173 for C. neoformans being unique to Thailand. Most of the C. gattii isolates were ST7 = VGIIb, which is identical to the less virulent minor Vancouver island outbreak genotype, indicating Thailand as a stepping stone in the global spread of this outbreak strain. The current study revealed a greater genetic diversity and a wider range of major molecular types being present amongst Thai cryptococcal isolates than previously reported. PMID:23861989

  18. isolation and characterisation of cryptococcus neoformans

    African Journals Online (AJOL)

    2014-08-01

    Aug 1, 2014 ... and Cryptococcus gattii from several environmental sites in Nairobi, Kenya .This could probably ... Globally, the risk of cryptococcal meningitis in HIV/. AIDS is estimated ..... gattii from the Amazon rainforest. PLoS One. 2013 ...

  19. Cryptococcal transmigration across a model brain blood-barrier: evidence of the Trojan horse mechanism and differences between Cryptococcus neoformans var. grubii strain H99 and Cryptococcus gattii strain R265.

    Science.gov (United States)

    Sorrell, Tania C; Juillard, Pierre-Georges; Djordjevic, Julianne T; Kaufman-Francis, Keren; Dietmann, Anelia; Milonig, Alban; Combes, Valery; Grau, Georges E R

    2016-01-01

    Cryptococcus neoformans (Cn) and Cryptococcus gattii (Cg) cause neurological disease and cross the BBB as free cells or in mononuclear phagocytes via the Trojan horse mechanism, although evidence for the latter is indirect. There is emerging evidence that Cn and the North American outbreak Cg strain (R265) more commonly cause neurological and lung disease, respectively. We have employed a widely validated in vitro model of the BBB, which utilizes the hCMEC/D3 cell line derived from human brain endothelial cells (HBEC) and the human macrophage-like cell line, THP-1, to investigate whether transport of dual fluorescence-labelled Cn and Cg across the BBB occurs within macrophages. We showed that phagocytosis of Cn by non-interferon (IFN)-γ stimulated THP-1 cells was higher than that of Cg. Although Cn and Cg-loaded THP-1 bound similarly to TNF-activated HBECs under shear stress, more Cn-loaded macrophages were transported across an intact HBEC monolayer, consistent with the predilection of Cn for CNS infection. Furthermore, Cn exhibited a higher rate of expulsion from transmigrated THP-1 compared with Cg. Our results therefore provide further evidence for transmigration of both Cn and Cg via the Trojan horse mechanism and a potential explanation for the predilection of Cn to cause CNS infection. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  20. MLST and Whole-Genome-Based Population Analysis of Cryptococcus gattii VGIII Links Clinical, Veterinary and Environmental Strains, and Reveals Divergent Serotype Specific Sub-populations and Distant Ancestors

    Science.gov (United States)

    Firacative, Carolina; Roe, Chandler C.; Malik, Richard; Ferreira-Paim, Kennio; Escandón, Patricia; Sykes, Jane E.; Castañón-Olivares, Laura Rocío; Contreras-Peres, Cudberto; Samayoa, Blanca; Sorrell, Tania C.; Castañeda, Elizabeth; Lockhart, Shawn R.; Engelthaler, David M.; Meyer, Wieland

    2016-01-01

    The emerging pathogen Cryptococcus gattii causes life-threatening disease in immunocompetent and immunocompromised hosts. Of the four major molecular types (VGI-VGIV), the molecular type VGIII has recently emerged as cause of disease in otherwise healthy individuals, prompting a need to investigate its population genetic structure to understand if there are potential genotype-dependent characteristics in its epidemiology, environmental niche(s), host range and clinical features of disease. Multilocus sequence typing (MLST) of 122 clinical, environmental and veterinary C. gattii VGIII isolates from Australia, Colombia, Guatemala, Mexico, New Zealand, Paraguay, USA and Venezuela, and whole genome sequencing (WGS) of 60 isolates representing all established MLST types identified four divergent sub-populations. The majority of the isolates belong to two main clades, corresponding either to serotype B or C, indicating an ongoing species evolution. Both major clades included clinical, environmental and veterinary isolates. The C. gattii VGIII population was genetically highly diverse, with minor differences between countries, isolation source, serotype and mating type. Little to no recombination was found between the two major groups, serotype B and C, at the whole and mitochondrial genome level. C. gattii VGIII is widespread in the Americas, with sporadic cases occurring elsewhere, WGS revealed Mexico and USA as a likely origin of the serotype B VGIII population and Colombia as a possible origin of the serotype C VGIII population. Serotype B isolates are more virulent than serotype C isolates in a murine model of infection, causing predominantly pulmonary cryptococcosis. No specific link between genotype and virulence was observed. Antifungal susceptibility testing against six antifungal drugs revealed that serotype B isolates are more susceptible to azoles than serotype C isolates, highlighting the importance of strain typing to guide effective treatment to improve the

  1. Molecular characterization and antifungal susceptibility of Cryptococcus neoformans strains collected from a single institution in Lima, Peru : Revista Iberoamericana De Micologia

    NARCIS (Netherlands)

    Bejar, V.; Tello, M.; Garcia, R.; Guevara, J. M.; Gonzales, S.; Vergaray, G.; Valencia, E.; Abanto, E.; Ortega-Loayza, A. G.; Hagen, F.; Gutierrez, E. L.

    2015-01-01

    Background: Cryptococcosis is a fungal infection with a worldwide distribution, mainly caused by Cryptococcus neoformans and Cryptococcus gattii. Aims: To molecularly characterize the mating-types, serotypes, genotypes and antifungal susceptibility profiles of a set of retrospectively isolated C.

  2. Cryptococcosis (C. gattii)

    Science.gov (United States)

    ... Foodborne, Waterborne, and Environmental Diseases Mycotic Diseases Branch C. gattii Infection Recommend on Facebook Tweet Share Compartir ... as British Columbia and the U.S. Pacific Northwest. C. gattii cryptococcosis is a rare infection that people ...

  3. Adaptive Immunity to Cryptococcus neoformans Infections

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    Liliane Mukaremera

    2017-11-01

    Full Text Available The Cryptococcus neoformans/Cryptococcus gattii species complex is a group of fungal pathogens with different phenotypic and genotypic diversity that cause disease in immunocompromised patients as well as in healthy individuals. The immune response resulting from the interaction between Cryptococcus and the host immune system is a key determinant of the disease outcome. The species C. neoformans causes the majority of human infections, and therefore almost all immunological studies focused on C. neoformans infections. Thus, this review presents current understanding on the role of adaptive immunity during C. neoformans infections both in humans and in animal models of disease.

  4. Sporothrix schenckii fungemia without disseminated sporotrichosis.

    OpenAIRE

    Kosinski, R M; Axelrod, P; Rex, J H; Burday, M; Sivaprasad, R; Wreiole, A

    1992-01-01

    Fungemia is a rare complication of Sporothrix schenckii infection and has always been associated with disseminated sporotrichosis. We describe an immunocompetent patient with localized lymphocutaneous sporotrichosis from whose blood the fungus was isolated. A lysis-centrifugation blood culture system may have improved our ability to detect low-level S. schenckii fungemia.

  5. Fungemia in a university hospital: an epidemiological approach Fungemia em hospital universitário: uma abordagem epidemiológica

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    Fabíola Maria Marques do Couto

    2011-12-01

    Full Text Available INTRODUCTION: Fungemia corresponds to the isolation of fungi in the bloodstream and occurs mostly in immunosuppressed patients. The early diagnosis and treatment of these infections are relevant given the serious threat to the affected patients and possible spread to other organs, often becoming fatal. The growing number of fungemia associated with poor prognosis resulted in this research aiming to diagnose and assess the epidemiological aspects of hematogenous infections by fungi. METHODS: The study included 58 blood samples collected within a 1-year period, from patients at the Hospital das Clinicas, Federal University of Pernambuco, by venipuncture in vacuum tubes. Blood samples were processed for direct examination and culture and identification, conducted by observing the macroscopic and microscopic characteristics, as well as physiological characteristics when necessary. RESULTS: Eight (13.8% episodes of fungemia were identified, accounting for the total sample, and these pathogens were Candida, Histoplasma, Trichosporon, Cryptococcus, and a dematiaceous fungus. C. albicans was the prevalent species, accounting for 37.5% of the cases. Most affected patients were adult males. There was no predominance for any activity, and the risk of acquired immunodeficiency syndrome was the underlying pathology most often cited. CONCLUSIONS: The isolation of fungi considered as emergent species, such as C. membranifaciens and dematiaceous species, highlights the importance of epidemiological monitoring of cases of fungemia in immunocompromised patients, as the therapy of choice depends on the knowledge of the aethiological agent.INTRODUÇÃO: Fungemia corresponde ao isolamento de fungos na corrente sanguínea e ocorre, sobretudo, em pacientes imunossuprimidos. O diagnóstico e tratamento precoce destas infecções são relevantes diante da grave ameaça aos pacientes acometidos e possível disseminação via hematogênica para outros órgãos, tornando

  6. Molecules at the interface of Cryptococcus and the host that determine disease susceptibility.

    Science.gov (United States)

    Wozniak, Karen L; Olszewski, Michal A; Wormley, Floyd L

    2015-05-01

    Cryptococcus neoformans and Cryptococcus gattii, the predominant etiological agents of cryptococcosis, are fungal pathogens that cause disease ranging from a mild pneumonia to life-threatening infections of the central nervous system (CNS). Resolution or exacerbation of Cryptococcus infection is determined following complex interactions of several host and pathogen derived factors. Alternatively, interactions between the host and pathogen may end in an impasse resulting in the establishment of a sub-clinical Cryptococcus infection. The current review addresses the delicate interaction between the host and Cryptococcus-derived molecules that determine resistance or susceptibility to infection. An emphasis will be placed on data highlighted at the recent 9th International Conference on Cryptococcus and Cryptococcosis (ICCC). Copyright © 2015. Published by Elsevier Inc.

  7. [Cyptococcus gattii isolated from a cheetah (Acinonyx jubatus) in the National Zoo of Cuba].

    Science.gov (United States)

    Polo Leal, Jorge Luis; Fernández Andreu, Carlos Manuel; Martínez Machín, Gerardo; Illnait Zaragozi, María Teresa; Perurena Lancha, Mayda Rosa

    2010-01-01

    Cryptoccosis--systemic mycosis caused by Cryptococcus species--has considerably raised its incidence in the last years, mainly associated with the human immunodeficiency virus infection. It has also been described in animals, but rare cases. a case of a female cheetah (Acinonyx jubatus) kept in the Nacional Zoo of Havana was presented. The animal came from South Africa. She began losing weight, and suffering asthenia, anorexia and breathing problems with abundant nasal secretion. mycological testing of these secretions disclosed the presence of serotype B Cryptococcus gattii. Because of the origin and captive condition of the animal, it was believed that the infection had been latent for 16 months at least. up to the present, in Cuba, all clinical Cryptococcus isolates were C. neoformans var. grubii, so it is considered that the infection was caught in the country of origin of the female cheetah. This is the first C. gattii isolate in Cuba from an animal coming from South Africa where this fungus is endemic.

  8. Diversity of DNA fingerprints in Cryptococcus neoformans.

    Science.gov (United States)

    Varma, A; Swinne, D; Staib, F; Bennett, J E; Kwon-Chung, K J

    1995-07-01

    DNA fingerprint patterns of 156 Cryptococcus neoformans isolates (26 AIDS patients, 46 non-AIDS patients, and 40 environmental sources) from both varieties (126 C. neoformans var. neoformans and 30 C. neoformans var. gattii isolates) and from seven countries were analyzed by using the DNA probe UT-4p. Nine and twelve distinct DNA fingerprint patterns were observed for isolates of the C. neoformans var. neoformans and var. gattii, respectively. No pattern was unique to AIDS patients, non-AIDS patients, or the environment. Pattern II was observed more often in non-AIDS patients (8 of 23) than in AIDS patients (0 of 25). Pattern V was the most prevalent pattern (42 of 82) in clinical and environmental isolates. Isolates from three AIDS patients in Burundi and Zaire exhibited patterns identical to each other but different from those of isolates collected from their houses (i.e., dust of floors, walls, etc.) or a nearby pigeon coop. DNA fingerprint stability was determined for 53 isolates from nine non-AIDS patients at different time intervals during 5 to 128 weeks of antifungal therapy. For eight patients, the fingerprint pattern was stable while the ninth may have had a mixed infection. Pattern II was observed in 4 of 9 patients, which is similar to 4 of 14 in other non-AIDS patients as reported here. In spite of the extensive pattern heterogeneity among 15 C. neoformans var. gattii isolates in Australia, the patterns observed in seven California isolates were quite different from those in Australia. Among isolates of C. neoformans var. gattii, one fingerprint pattern (designated b) was observed in several countries of the Far East. The fingerprint patterns of two of three environmental isolates from Eucalyptus camaldulensis trees in Australia were identical to those of 2 of the 12 clinical isolates from the country.

  9. Environmental isolation, biochemical identification, and antifungal drug susceptibility of Cryptococcus species

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    Valter Luis Iost Teodoro

    2013-12-01

    Full Text Available Introduction The incidence of opportunistic fungal infections has increased in recent years and is considered an important public health problem. Among systemic and opportunistic mycoses, cryptococcosis is distinguished by its clinical importance due to the increased risk of infection in individuals infected by human immunodeficiency virus. Methods To determine the occurrence of pathogenic Cryptococcus in pigeon excrement in the City of Araraquara, samples were collected from nine environments, including state and municipal schools, abandoned buildings, parks, and a hospital. The isolates were identified using classical tests, and susceptibility testing for the antifungal drugs (fluconazole, itraconazole, voriconazole, and amphotericin B independently was also performed. After collection, the excrement samples were plated on Niger agar and incubated at room temperature. Results A total of 87 bird dropping samples were collected, and 66.6% were positive for the genus Cryptococcus. The following species were identified: Cryptococcus neoformans (17.2%, Cryptococcus gattii (5.2%, Cryptococcus ater (3.5%, Cryptococcus laurentti (1.7%, and Cryptococcus luteolus (1.7%. A total of 70.7% of the isolates were not identified to the species level and are referred to as Cryptococcus spp. throughout the manuscript. Conclusions Although none of the isolates demonstrated resistance to antifungal drugs, the identification of infested areas, the proper control of birds, and the disinfection of these environments are essential for the epidemiological control of cryptococcosis.

  10. Saccharomyces cerevisiae var. boulardii fungemia following probiotic treatment

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    Marcelo C. Appel-da-Silva

    2017-12-01

    Full Text Available Probiotics are commonly prescribed as an adjuvant in the treatment of antibiotic-associated diarrhea caused by Clostridium difficile. We report the case of an immunocompromised 73-year-old patient on chemotherapy who developed Saccharomyces cerevisiae var. boulardii fungemia in a central venous catheter during treatment of antibiotic-associated pseudomembranous colitis with the probiotic Saccharomyces cerevisiae var. boulardii. Fungemia was resolved after interruption of probiotic administration without the need to replace the central venous line. Keywords: Saccharomyces, Probiotics, Fungemia, Critical illness, Clostridium difficile

  11. Cryptococcus Neoformans: The First Reported Case of Isolation in Pigeon Excreta in the City of Votuporanga-São Paulo

    Directory of Open Access Journals (Sweden)

    Cátia Rezende

    2015-12-01

    Full Text Available Cryptococcocus neoformans and Cryptococcus gattii are closely related pathogenic fungi that cause pneumonia and meningitis in both immunocompromised and immunocompetent hosts. Both species are found in the environment, and are acquired via inhalation, leading to an initial pulmonary infection. We evaluated 48 air samples and 32 samples of pigeon excreta from June to August, 2007. The presence of capsule was examined with Indian ink preparation. They were also tested for urease and phenoloxidase enzymes. Cryptococcus neoformans was recovered from pigeon excreta in 3.1%. The results suggest that climatic conditions can affect the occurrence of the yeast in different environmental sources.

  12. Plants promote mating and dispersal of the human pathogenic fungus Cryptococcus.

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    Deborah J Springer

    Full Text Available Infections due to Cryptococcus are a leading cause of fungal infections worldwide and are acquired as a result of environmental exposure to desiccated yeast or spores. The ability of Cryptococcus to grow, mate, and produce infectious propagules in association with plants is important for the maintenance of the genetic diversity and virulence factors important for infection of animals and humans. In the Western United States and Canada, Cryptococcus has been associated with conifers and tree species other than Eucalyptus; however, to date Cryptococcus has only been studied on live Arabidopsis thaliana, Eucalyptus sp., and Terminalia catappa (almond seedlings. Previous research has demonstrated the ability of Cryptococcus to colonize live plants, leaves, and vasculature. We investigated the ability of Cryptococcus to grow on live seedlings of the angiosperms, A. thaliana, Eucalyptus camaldulensis, Colophospermum mopane, and the gymnosperms, Pseudotsuga menziesii (Douglas fir, and Tsuga heterophylla (Western hemlock. We observed a broad-range ability of Cryptococcus to colonize both traditional infection models as well as newly tested conifer species. Furthermore, C. neoformans, C. deneoformans, C. gattii (VGI, C. deuterogattii (VGII and C. bacillisporus (VGIII were able to colonize live plant leaves and needles but also undergo filamentation and mating on agar seeded with plant materials or in saprobic association with dead plant materials. The ability of Cryptococcus to grow and undergo filamentation and reproduction in saprobic association with both angiosperms and gymnosperms highlights an important role of plant debris in the sexual cycle and exposure to infectious propagules. This study highlights the broad importance of plants (and plant debris as the ecological niche and reservoirs of infectious propagules of Cryptococcus in the environment.

  13. Saccharomyces cerevisiae var. boulardii fungemia following probiotic treatment

    OpenAIRE

    Appel-da-Silva, Marcelo C.; Narvaez, Gabriel A.; Perez, Leandro R.R.; Drehmer, Laura; Lewgoy, Jairo

    2017-01-01

    Probiotics are commonly prescribed as an adjuvant in the treatment of antibiotic-associated diarrhea caused by Clostridium difficile. We report the case of an immunocompromised 73-year-old patient on chemotherapy who developed Saccharomyces cerevisiae var. boulardii fungemia in a central venous catheter during treatment of antibiotic-associated pseudomembranous colitis with the probiotic Saccharomyces cerevisiae var. boulardii. Fungemia was resolved after interruption of probiotic administrat...

  14. Asociacion de leveduras del genero Cryptococcus con especies de Eucalyptus en Santafe de Bogota Isolation of Cryptococcus sp. associated with Eucalyptus trees in Santafé de Bogota

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    A. Duarte

    1994-04-01

    Full Text Available El aislamiento de Cryptococcus neoformans var. gattii, serotipo B, a partir del medio ambiente se estableció inicialmente en Australia en 1989, en asocio con el Eucalyptus camaldulensis y posteriormente con E. tereticornis. Con estos hallazgos se postuló que desde allí, el hongo se ha podido exportar, por medio de las semillas contaminadas, a otras regiones geográficas, incluyendo Colombia. El objetivo de éste estudio fue identificar las levaduras del género Cryptococcus asociadas con especies de Eucalyptus sp., como primera evaluación en la ecología de C. neoformans var. gattii en nuestro país. Se realizó en Santafé de Bogotá, con una población de 100 árboles ubicados al centro, nororiente, oriente y occidente de la ciudad, recolectando de cada uno de ellos flores, frutos, hojas, cortezas y detritos; el procesamiento de las muestras incluyó extracción del material con una solución salina con antibióticos, siembra en medios selectivos e identificación de las especies con base en las características morfológicas, macro y microscópicas y bioquímicas. Se aislaron 27 cepas de Cryptococcus pertenecientes a 9 especies de Cryptococcus, a partir de 21 árboles ubicados en 5 zonas diferentes de la ciudad. Se aisló C. neoformans y se identificó como C. neoformans var. neoformans serotipo A. Estos datos iniciales son importantes como primera evaluación de la asociación de Cryptococcus sp. con los Eucalyptus en nuestro país.Environmental isolation of Cryptococcus neoformans var. gattii was first made in Australia in 1989 by ELLIS. He established a specific association with the tree species Eucalyptus camaldulensis and E. tereticornis. Based on his findings, ELLIS proposed that the fungus could be exported from Australia to others regions, including Colombia, by means of infected seeds. The purpose of this study was to isolate and identify Cryptococcus sp., associated with Eucalyptus trees; this is the first ecological evaluation

  15. Isolation and molecular characterization of Cryptococcus species isolated from pigeon nests and Eucalyptus trees.

    Science.gov (United States)

    Kamari, A; Sepahvand, A; Mohammadi, R

    2017-06-01

    Cryptococcus species are pathogenic and non-pathogenic basidiomycete yeasts that are found widely in the environment. Based on phenotypic methods, this genus has many species; however, its taxonomy is presently being re-evaluated by modern techniques. The Cryptococcus species complex includes two sibling taxa of Cryptococcus neoformans and Cryptococcus gattii . We aimed to investigate the possible distribution of Cryptococcus species in pigeon nests and Eucalyptus trees in Ilam, Iran, using molecular techniques. Two hundred and seventy-four specimens were collected from pigeon nests and Eucalyptus trees during 2016-2017. All the specimens were sub-cultured on Sabouraud Glucose Agar with chloramphenicol and bird seed agar. For molecular identification, the ITS15.8SITS2 rDNA region was amplified using the first and fourth internal transcribed spacer (ITS1 and ITS4, respectively) primers. The purified products were applied for cycle sequencing reactions in forward direction with ITS1 primer. The obtained results were analyzed with Chromas 2.3. Thirty-three out of 186 cultures (17.7%) and 11 out of 88 cultures (12.5%) were positive among pigeon nest and Eucalyptus tree specimens, respectively. Cryptococcus albidus (17.2%), C. albidus var. kuetzingii (3.4%), C. adeliensis (3.4%), C. uzbekistanensis (3.4%), and C. neoformans var. grubii (3.4%) were isolated from pigeon nests, and Cryptococcus adeliensis (25%) was the only Cryptococcus species isolated from Eucalyptus trees. The presence of pigeons and Eucalyptus trees in the vicinity of some particular places such as rest homes and hospitals should be considered as a risk factor for the immunocompromised population.

  16. Rhodotorula minuta fungemia in a ewe lamb.

    Science.gov (United States)

    Chitko-McKown, C G; Leymaster, K A; Heaton, M P; Griffin, D D; Veatch, J K; Jones, S A; Clawson, M L

    2014-12-01

    An 8-month-old crossbred ewe, normal upon physical examination, was humanely euthanized for tissue collection. After approximately 3 weeks in tissue culture, fungi began budding out of cells obtained from the choroid plexus. After an additional 3 weeks, budding was observed in kidney cell cultures and eventually in monocyte cultures as well. Serum from the lamb was submitted to the Veterinary Diagnostic Laboratory at Colorado State University for fungal diagnosis and was found negative for Aspergillus, Blastomyces, Coccidioidomycosis and Histoplasmosis. DNA was isolated from fungi collected from tissue culture supernatants and used in a set of pan-fungal PCR assays with DNA from Candida acting as a positive control. PCR products were sequenced and BLAST analysis performed. The unknown fungal sequence aligned with 100% identity to Rhodotorula minuta an emerging opportunistic pathogen. Samples were submitted to The Fungal Testing Laboratory at The University of Texas Health Science Center at San Antonio for additional validation. We believe this to be the first report of Rhodotorula fungemia in a sheep in the United States. © 2013 Blackwell Verlag GmbH.

  17. Central Venous Catheter-related Fungemia Caused by Rhodotorula glutinis.

    Science.gov (United States)

    Miglietta, Fabio; Letizia Faneschi, Maria; Braione, Adele; Palumbo, Claudio; Rizzo, Adriana; Lobreglio, Giambattista; Pizzolante, Maria

    2015-01-01

    Bloodstream infection due to Rhodotorula glutinis is extremely rare and mostly associated with underlying immunosuppression or cancer. Vascular access devices provide the necessary surfaces for biofilm formation and are currently responsible for a significant percentage of human infections. In this work, we describe a rare case of central venous catheter-related Rhodotorula glutinis fungemia in a female patient with acute myelogenous leukemia in remission. The timely removal of central venous catheter was an essential element for overcoming this CVC-related Rhodotorula fungemia.

  18. National Surveillance of fungemia in Denmark (2004 to 2009)

    DEFF Research Database (Denmark)

    Arendrup, Maiken Cavling; Bruun, Brita; Christensen, Jens Jørgen

    2011-01-01

    to fluconazole (MIC>4 μg/ml) occurred in: C. albicans 7/1183 (0.6%), C. dubliniensis 2/65 (3.1%), C. parapsilosis 5/83 (6.0%) and C. tropicalis 7/104 (6.7%). Overall, 70.8% of fungemia isolates were fully fluconazole susceptible but the proportion decreased (79.7% to 68.9%, P=0.02). The study confirmed a three...... times higher incidence rate of fungemia in Denmark compared to other Nordic countries and identified marked differences related to age and gender. Decreased susceptibility to fluconazole was frequent and increasing....

  19. The mating type locus (MAT and sexual reproduction of Cryptococcus heveanensis: insights into the evolution of sex and sex-determining chromosomal regions in fungi.

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    Banu Metin

    2010-05-01

    Full Text Available Mating in basidiomycetous fungi is often controlled by two unlinked, multiallelic loci encoding homeodomain transcription factors or pheromones/pheromone receptors. In contrast to this tetrapolar organization, Cryptococcus neoformans/Cryptococcus gattii have a bipolar mating system, and a single biallelic locus governs sexual reproduction. The C. neoformans MAT locus is unusually large (>100 kb, contains >20 genes, and enhances virulence. Previous comparative genomic studies provided insights into how this unusual MAT locus might have evolved involving gene acquisitions into two unlinked loci and fusion into one contiguous locus, converting an ancestral tetrapolar system to a bipolar one. Here we tested this model by studying Cryptococcus heveanensis, a sister species to the pathogenic Cryptococcus species complex. An extant sexual cycle was discovered; co-incubating fertile isolates results in the teleomorph (Kwoniella heveanensis with dikaryotic hyphae, clamp connections, septate basidia, and basidiospores. To characterize the C. heveanensis MAT locus, a fosmid library was screened with C. neoformans/C. gattii MAT genes. Positive fosmids were sequenced and assembled to generate two large probably unlinked MAT gene clusters: one corresponding to the homeodomain locus and the other to the pheromone/receptor locus. Strikingly, two divergent homeodomain genes (SXI1, SXI2 are present, similar to the bE/bW Ustilago maydis paradigm, suggesting one or the other homeodomain gene was recently lost in C. neoformans/C. gattii. Sequencing MAT genes from other C. heveanensis isolates revealed a multiallelic homeodomain locus and at least a biallelic pheromone/receptor locus, similar to known tetrapolar species. Taken together, these studies reveal an extant C. heveanensis sexual cycle, define the structure of its MAT locus consistent with tetrapolar mating, and support the proposed evolutionary model for the bipolar Cryptococcus MAT locus revealing

  20. Malassezia pachydermatis fungemia in an adult with multibacillary leprosy

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    Jorge Roman

    2016-06-01

    Full Text Available Malassezia pachydermatis is a relatively rare agent of bloodstream infections. We describe an unusual case of Malassezia fungemia in an adult patient hospitalized for Staphylococcus aureus bacteremia who was also found to have multibacillary leprosy. Treatment of the patient required extensive medical management but resulted in a good outcome.

  1. Trichosporon fungemia in a pediatric patient with acute lymphoblastic leukemia

    OpenAIRE

    Luke Maxfield; Juliana J. Matthews; David Ryan Ambrosetti; Issa E. Ephtimios

    2015-01-01

    Trichosporon fungemia is a life-threatening opportunistic infection that is increasing in frequency. Invasive disease occurs almost exclusively in immunocompromised hosts, particularly in neutropenic adults with hematological malignancies and uncommonly in children. We report the case of a pediatric patient where disseminated trichosporonosis progressed while on micafungin, between treatments with voriconazole and amphotericin B, demonstrating the difficulty with and importance of prolonged a...

  2. Cryptococcus: isolamento ambiental e caracterização bioquímica

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    E.C. Araújo Júnior

    2015-08-01

    Full Text Available O gênero Cryptococcus caracteriza-se por ser uma levedura responsável por infecção sistêmica, causada pelas espécies Cryptococcus neoformans e Cryptococcus gattii. O fungo é encontrado em substratos de origem animal e vegetal, e a infecção ocorre com a inalação de basidiósporos ou leveduras desidratadas infectantes presentes no ambiente. O presente trabalho teve por objetivo pesquisar a existência de microfocos de Cryptococcussp.em amostras ambientais da cidade de Araçatuba, São Paulo, com a finalidade de minimizar os riscos de contaminação do homem e dos animais, buscando o conhecimento da ecoepidemiologia do Cryptococcus. Foram colhidas 50 amostras oriundas de ocos e troncos de árvores (Cassiasp., Ficussp., Caesalpinea peltophorides de 10 locais representativos do perímetro urbano, as quais foram encaminhadas ao Laboratório de Bacteriologia e Micologia da Faculdade de Medicina Veterinária de Araçatuba-Unesp, onde foram processadas e semeadas em placas de Petri contendo ágar semente de Níger e Sabouraud dextrose com clorafenicol e incubadas à temperatura de 30ºC, por um período não inferior a cinco dias. Posteriormente, foram submetidas às provas bioquímicas: produção de urease, termotolerância a 37ºC e quimiotipagem em ágar CGB (L-canavanina-glicina-azul de bromotimol. A análise dos resultados revelaram que 17 (34% dos cultivos foram positivos para o gênero Cryptococcus, sendo nove (18% para Cryptococcus gattiie oito (16% para Cryptococcus neoformans. Outras leveduras correlacionadas, como Rhodotorula sp. e Candida sp., também foram isoladas. Conclui-se que os basidiósporos de Cryptococcusencontram-se dispersos na natureza, constituindo microfocos ambientais, não vinculados necessariamente a um único hospedeiro.

  3. Seminational surveillance of fungemia in Denmark: Notably high rates of fungemia and numbers of isolates with reduced azole susceptibility

    DEFF Research Database (Denmark)

    Arendrup, M.C.; Fuursted, K.; Gahrn-Hansen, B.

    2005-01-01

    The aim of this study was to present the first set of comprehensive data on fungemia in Denmark including the distribution of species and range of susceptibility to major antifungal compounds based on a seminational surveillance study initiated in 2003. The catchment area of the participating hos...

  4. [Preservation of high risk fungal cultures of Histoplasma and Cryptococcus].

    Science.gov (United States)

    Fernández Andreu, C Carlos Manuel; Díaz Suárez, Luis Alberto; Ilnait Zaragozi, María Teresa; Aragonés López, Carlos; Martínez Machín, Gerardo; Perurena Lancha, Mayda R

    2012-01-01

    culture collections are responsible for providing the microbial resources for development of biological sciences. Storage in distilled water is one of the easiest and least expensive method for long-term fungal preservation. to evaluate the usefulness of this preservation method in fungal culture of Histoplasma and Cryptococcus. the preservation condition of the highest biological risk species from Histoplasma y Cryptococcus genera, included in the fungal culture collection of "Pedro Kouri" Institute of Tropical Medicine in Havana, was evaluated in this study. One hundred and two strains stored in distilled water, 92% of which had been preserved for more than 10 years, were analyzed. the percentages of recovered strains from H. capsulatum, C. neoformans and C. gattii were 64.3%; 79.1% and 100% respectively. This method of preservation proved to be satisfactory for fungal culture in labs with limited financial resources. A web-based database with interesting information about the collection was made. The importance of strict compliance with the biosafety measures in these collections, particularly with high risk pathogens. preservation of fungal cultures in distilled water is a very useful method for laboratories with limited resources. Culture collections should be assumed as an essential activity in order to solve increasing challenges in the development of biomedical sciences.

  5. Characterization of the antigenicity of Cpl1, a surface protein of Cryptococcus neoformans var. neoformans.

    Science.gov (United States)

    Cai, Jian-Piao; Liu, Ling-Li; To, Kelvin K W; Lau, Candy C Y; Woo, Patrick C Y; Lau, Susanna K P; Guo, Yong-Hui; Ngan, Antonio H Y; Che, Xiao-Yan; Yuen, Kwok-Yung

    2015-01-01

    Cryptococcus neoformans var. neoformans is an important fungal pathogen. The capsule is a well established virulence factor and a target site for diagnostic tests. The CPL1 gene is required for capsular formation and virulence. The protein product Cpl1 has been proposed to be a secreted protein, but the characteristics of this protein have not been reported. Here we sought to characterize Cpl1. Phylogenetic analysis showed that the Cpl1 of C. neoformans var. neoformans and the Cpl1 orthologs identified in C. neoformans var. grubii and C. gattii formed a distinct cluster among related fungi; while the putative ortholog found in Trichosporon asahii was distantly related to the Cryptococcus cluster. We expressed Cpl1 abundantly as a secreted His-tagged protein in Pichia pastoris. The protein was used to immunize guinea pigs and rabbits for high titer mono-specific polyclonal antibody that was shown to be highly specific against the cell wall of C. neoformans var. neoformans and did not cross react with C. gattii, T. asahii, Aspergillus spp., Candida spp. and Penicillium spp. Using the anti-Cpl1 antibody, we detected Cpl1 protein in the fresh culture supernatant of C. neoformans var. neoformans and we showed by immunostaining that the Cpl1 protein was located on the surface. The Cpl1 protein is a specific surface protein of C. neoformans var. neoformans. © 2015 by The Mycological Society of America.

  6. Cryptococcus spp isolated from dust microhabitat in Brazilian libraries

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    Leite Diniz P

    2012-06-01

    Full Text Available Abstract Background The Cryptococcus spp is currently composed of encapsulated yeasts of cosmopolitan distribution, including the etiological agents of cryptococcosis. The fungus are found mainly in substrates of animal and plant origin. Human infection occurs through inhalation of spores present in the environment. Methods Eighty-four swab collections were performed on dust found on books in three libraries in the city of Cuiabá, state of Mato Grosso, Brazil. The material was seeded in Sabouraud agar and then observed for characteristics compatible with colonies with a creamy to mucous aspect; the material was then isolated in birdseed (Niger agar and cultivated at a temperature of 37°C for 5 to 7 days. Identification of isolated colonies was performed by microscopic observation in fresh preparations dyed with India ink, additional tests performed on CGB (L-canavanine glycine bromothymol blue, urea broth, and carbohydrate assimilation tests (auxanogram. Results Of the 84 samples collected from book dust, 18 (21.4% were positive for Cryptococcus spp totalizing 41 UFC’s. The most frequently isolated species was C. gattii 15 (36.6%; followed by C. terreus, 12 (29.3%; C. luteolus 4 (9.8%; C. neoformans, and C. uniguttulatus 3 (7.3%, and C. albidus and C. humiculus with 2 (4.6% of the isolates. Conclusion The high biodiversity of the yeasts of the Cryptococcus genus, isolated from different environmental sources in urban areas of Brazil suggests the possibility of individuals whose immune systems have been compromised or even healthy individuals coming into sources of fungal propagules on a daily bases throughout their lives. This study demonstrates the acquisition possible of cryptococcosis infection from dust in libraries.

  7. Cryptococcus spp isolated from dust microhabitat in Brazilian libraries.

    Science.gov (United States)

    Leite, Diniz P; Amadio, Janaina V R S; Martins, Evelin R; Simões, Sara A A; Yamamoto, Ana Caroline A; Leal-Santos, Fábio A; Takahara, Doracilde T; Hahn, Rosane C

    2012-06-08

    The Cryptococcus spp is currently composed of encapsulated yeasts of cosmopolitan distribution, including the etiological agents of cryptococcosis. The fungus are found mainly in substrates of animal and plant origin. Human infection occurs through inhalation of spores present in the environment. Eighty-four swab collections were performed on dust found on books in three libraries in the city of Cuiabá, state of Mato Grosso, Brazil. The material was seeded in Sabouraud agar and then observed for characteristics compatible with colonies with a creamy to mucous aspect; the material was then isolated in birdseed (Niger) agar and cultivated at a temperature of 37°C for 5 to 7 days. Identification of isolated colonies was performed by microscopic observation in fresh preparations dyed with India ink, additional tests performed on CGB (L-canavanine glycine bromothymol blue), urea broth, and carbohydrate assimilation tests (auxanogram). Of the 84 samples collected from book dust, 18 (21.4%) were positive for Cryptococcus spp totalizing 41 UFC's. The most frequently isolated species was C. gattii 15 (36.6%); followed by C. terreus, 12 (29.3%); C. luteolus 4 (9.8%); C. neoformans, and C. uniguttulatus 3 (7.3%), and C. albidus and C. humiculus with 2 (4.6%) of the isolates. The high biodiversity of the yeasts of the Cryptococcus genus, isolated from different environmental sources in urban areas of Brazil suggests the possibility of individuals whose immune systems have been compromised or even healthy individuals coming into sources of fungal propagules on a daily bases throughout their lives. This study demonstrates the acquisition possible of cryptococcosis infection from dust in libraries.

  8. Rhodotorula fungemia: two cases and a brief review.

    Science.gov (United States)

    Duggal, Shalini; Jain, Hemant; Tyagi, Amit; Sharma, Anuradha; Chugh, T D

    2011-11-01

    Rhodotorula is emerging as an important cause of nosocomial and opportunistic infections. We present two cases of Rhodotorula mucilaginosa fungemia diagnosed over a period of 3 months at our hospital. The first case was of a pre-term neonate in the neonatal ICU who presented with respiratory failure and sepsis. The second involved an adult female who had been injured in a road traffic accident requiring an operation for a hematoma and was later shifted to the medical ICU. For a new hospital like ours, finding two cases of Rhodotorula fungemia within a span of 3 months prompted us to describe them in this report. These cases emphasize the emerging importance of Rhodotorula mucilaginosa as a pathogen and the importance of identification and MIC testing for all fungal isolates recovered from the blood stream.

  9. Discovery of a modified tetrapolar sexual cycle in Cryptococcus amylolentus and the evolution of MAT in the Cryptococcus species complex.

    Directory of Open Access Journals (Sweden)

    Keisha Findley

    Full Text Available Sexual reproduction in fungi is governed by a specialized genomic region called the mating-type locus (MAT. The human fungal pathogenic and basidiomycetous yeast Cryptococcus neoformans has evolved a bipolar mating system (a, α in which the MAT locus is unusually large (>100 kb and encodes >20 genes including homeodomain (HD and pheromone/receptor (P/R genes. To understand how this unique bipolar mating system evolved, we investigated MAT in the closely related species Tsuchiyaea wingfieldii and Cryptococcus amylolentus and discovered two physically unlinked loci encoding the HD and P/R genes. Interestingly, the HD (B locus sex-specific region is restricted (∼2 kb and encodes two linked and divergently oriented homeodomain genes in contrast to the solo HD genes (SXI1α, SXI2a of C. neoformans and Cryptococcus gattii. The P/R (A locus contains the pheromone and pheromone receptor genes but has expanded considerably compared to other outgroup species (Cryptococcus heveanensis and is linked to many of the genes also found in the MAT locus of the pathogenic Cryptococcus species. Our discovery of a heterothallic sexual cycle for C. amylolentus allowed us to establish the biological roles of the sex-determining regions. Matings between two strains of opposite mating-types (A1B1×A2B2 produced dikaryotic hyphae with fused clamp connections, basidia, and basidiospores. Genotyping progeny using markers linked and unlinked to MAT revealed that meiosis and uniparental mitochondrial inheritance occur during the sexual cycle of C. amylolentus. The sexual cycle is tetrapolar and produces fertile progeny of four mating-types (A1B1, A1B2, A2B1, and A2B2, but a high proportion of progeny are infertile, and fertility is biased towards one parental mating-type (A1B1. Our studies reveal insights into the plasticity and transitions in both mechanisms of sex determination (bipolar versus tetrapolar and sexual reproduction (outcrossing versus inbreeding with

  10. Trichosporon fungemia in a pediatric patient with acute lymphoblastic leukemia

    Directory of Open Access Journals (Sweden)

    Luke Maxfield

    2015-01-01

    Full Text Available Trichosporon fungemia is a life-threatening opportunistic infection that is increasing in frequency. Invasive disease occurs almost exclusively in immunocompromised hosts, particularly in neutropenic adults with hematological malignancies and uncommonly in children. We report the case of a pediatric patient where disseminated trichosporonosis progressed while on micafungin, between treatments with voriconazole and amphotericin B, demonstrating the difficulty with and importance of prolonged and continuous treatment.

  11. Fungemia Due to Fusarium sacchari in an Immunosuppressed Patient

    Science.gov (United States)

    Guarro, Josep; Nucci, Marcio; Akiti, Tiyomi; Gené, Josepa; Barreiro, M. Da Gloria C.; Gonçalves, Renato T.

    2000-01-01

    The fungus Fusarium sacchari was isolated repeatedly from the blood of an immunosuppressed host. The infection was treated successfully with a small dose of amphotericin B. The strain was resistant to this antifungal in vitro. MICs and minimum fungicidal concentrations of six antifungals for the clinical isolate are provided. To our knowledge, this is the first report involving this fungus in a case of fungemia. PMID:10618130

  12. Ecological surveys of the Cryptococcus species complex in China.

    Science.gov (United States)

    Li, An-Sheng; Pan, Wei-Hua; Wu, Shao-Xi; Hideaki, Taguchi; Guo, Ning-Ru; Shen, Yong-Nian; Lu, Gui-Xia; Pan, Ru-Gui; Zhu, Miao-Chang; Chen, Min; Shi, Wei-Ming; Liao, Wan-Qing

    2012-02-01

    Despite recent reports on the molecular epidemiology of cryptococcal infections in China, clinical isolates have been mostly reported from human immunodeficiency virus (HIV)-negative patients, and environmental isolates from China have rarely been included. The aim of this study was to investigate the ecological profile of Cryptococcus (C.) neoformans and C. gattii in China. A survey was performed in 10 cities from 20°N (North latitude) to 50°N and in a Eucalyptus (E.) camaldulensis forestry farm at the Guixi forestry center, China. Six hundred and twenty samples of pigeon droppings from 10 cities and 819 E. camaldulensis tree samples were collected and inoculated on caffeic acid cornmeal agar (CACA). The brown-colored colonies were recultured to observe their morphology, growth on canavanine-glycine-bromothymol-blue (CGB) medium, phenol oxidase and urease activities, serotype and mating type. There were obvious differences in the positive sample rates of C. neoformans in pigeon droppings collected from the different cities, ranging from 50% in the cities located at latitudes from 30°N - 40°N, 29% at 20°N - 30°N and 13% at 40°N - 50°N. There were no differences in positive bevy rates (approximately 80%) among the three grouped cities. Mycological tests of 101 isolates purified from pigeon droppings revealed that they were C. neoformans var. grubii. We also observed variable capsular size around the C. neoformans cells in colonies with variable melanin production and the bio-adhesion of the natural C. neoformans cells with other microorganisms. One urease-negative C. neoformans isolate was isolated from pigeon droppings in Jinan city. No C. gattii was isolated in this study.

  13. Repurposing of Aspirin and Ibuprofen as Candidate Anti-Cryptococcus Drugs

    Science.gov (United States)

    Ogundeji, Adepemi O.; Pohl, Carolina H.

    2016-01-01

    The usage of fluconazole and amphotericin B in clinical settings is often limited by, among other things, drug resistance development and undesired side effects. Thus, there is a constant need to find new drugs to better manage fungal infections. Toward this end, the study described in this paper considered the repurposing of aspirin (acetylsalicylic acid) and ibuprofen as alternative drugs to control the growth of cryptococcal cells. In vitro susceptibility tests, including a checkerboard assay, were performed to assess the response of Cryptococcus neoformans and Cryptococcus gattii to the above-mentioned anti-inflammatory drugs. Next, the capacity of these two drugs to induce stress as well as their mode of action in the killing of cryptococcal cells was determined. The studied fungal strains revealed a response to both aspirin and ibuprofen that was dose dependent, with ibuprofen exerting greater antimicrobial action. More importantly, the MICs of these drugs did not negatively (i) affect growth or (ii) impair the functioning of macrophages; rather, they enhanced the ability of these immune cells to phagocytose cryptococcal cells. Ibuprofen was also shown to act in synergy with fluconazole and amphotericin B. The treatment of cryptococcal cells with aspirin or ibuprofen led to stress induction via activation of the high-osmolarity glycerol (HOG) pathway, and cell death was eventually achieved through reactive oxygen species (ROS)-mediated membrane damage. The presented data highlight the potential clinical application of aspirin and ibuprofen as candidate anti-Cryptococcus drugs. PMID:27246782

  14. Recent and innovative methods for detection of bacteremia and fungemia

    International Nuclear Information System (INIS)

    Reller, L.B.

    1983-01-01

    Advances continue to be made in methods for more reliable or more rapid means of detecting bacteremia and fungemia. The importance of blood sample volume and broth dilution has been established in controlled studies. New technology includes the use of resins that remove antimicrobials from blood samples, detection of radioactivity from organisms given radiolabeled substrate, use of dyes that stain microbial DNA and RNA, use of slides coated with growth media, and lysis-centrifugation for trapping microorganisms. Technology now being considered includes counterimmunoelectrophoresis, head-space gas chromatography, electrical impedance, microcalorimetry, and the use of lasers to detect pH changes and turbidity

  15. Diagnostic issues, clinical characteristics, and outcomes for patients with fungemia

    DEFF Research Database (Denmark)

    Arendrup, Maiken Cavling; Sulim, Sofia; Holm, Anette

    2011-01-01

    This study investigated microbiological, clinical, and management issues and outcomes for Danish fungemia patients. Isolates and clinical information were collected at six centers. A total of 334 isolates, 316 episodes, and 305 patients were included, corresponding to 2/3 of the national episodes...... needed longer incubation. Species distribution varied by age, prior antifungal treatment (57% occurrence of C. glabrata, Saccharomyces cerevisiae, or C. krusei in patients with prior antifungal treatment versus 28% occurrence in those without it; P = 0.007), and clinical specialty (61% occurrence of C...

  16. Diagnostic Issues, Clinical Characteristics, and Outcomes for Patients with Fungemia

    DEFF Research Database (Denmark)

    Arendrup, Maiken Cavling; Sulim, Sofia; Holm, Anette

    2011-01-01

    This study investigated microbiological, clinical, and management issues and outcomes for Danish fungemia patients. Isolates and clinical information were collected at six centers. A total of 334 isolates, 316 episodes, and 305 patients were included, corresponding to 2/3 of the national episodes...... needed longer incubation. Species distribution varied by age, prior antifungal treatment (57% occurrence of C. glabrata, Saccharomyces cerevisiae, or C. krusei in patients with prior antifungal treatment versus 28% occurrence in those without it; P = 0.007), and clinical specialty (61% occurrence of C...

  17. Role of Sterylglucosidase 1 (Sgl1) on the pathogenicity of Cryptococcus neoformans: potential applications for vaccine development.

    Science.gov (United States)

    Rella, Antonella; Mor, Visesato; Farnoud, Amir M; Singh, Ashutosh; Shamseddine, Achraf A; Ivanova, Elitza; Carpino, Nicholas; Montagna, Maria T; Luberto, Chiara; Del Poeta, Maurizio

    2015-01-01

    Cryptococcosis caused by Cryptococcus neoformans and Cryptococcus gattii affects a large population and is a cause of significant morbidity and mortality. Despite its public health burden, there are currently no vaccines against cryptococcosis and new strategies against such infections are needed. In this study, we demonstrate that C. neoformans has the biochemical ability to metabolize sterylglucosides (SGs), a class of immunomodulatory glycolipids. Genetic manipulations that eliminate cryptococccal sterylglucosidase lead to the accumulation of SGs and generate a mutant strain (Δsgl1) that is non-pathogenic in the mouse models of cryptococcosis. Interestingly, this mutant strain acts as a vaccine strain and protects mice against cryptococcosis following infection with C. neoformans or C. gattii. The immunity induced by the Δsgl1 strain is not CD4(+) T-cells dependent. Immunocompromised mice, which lack CD4(+) T-cells, are able to control the infection by Δsgl1 and acquire immunity against the challenge by wild-type C. neoformans following vaccination with the Δsgl1 strain. These findings are particularly important in the context of HIV/AIDS immune deficiency and suggest that the Δsgl1 strain might provide a potential vaccination strategy against cryptococcosis.

  18. Rhodotorula mucilaginosa Fungemia and Pleural Tuberculosis in an Immunocompetent Patient: An Uncommon Association.

    Science.gov (United States)

    Pereira, Cláudia; Ribeiro, Sofia; Lopes, Virgínia; Mendonça, Teresa

    2016-02-01

    We report an atypical case of Rhodotorula mucilaginosa fungemia coexisting with pleural tuberculosis, in an immunocompetent host. The patient was an inhaled drug abuser and worked in a fruit market. The diagnosis of Rhodotorula mucilaginosa infection was established by the isolation of the yeast in two blood cultures followed by a good response to amphotericin B treatment. Persistent evening fever and pleural effusion led to the second diagnosis-pleural tuberculosis. In the last 5 years, this was the only case of Rhodotorula mucilaginosa fungemia in our hospital and the first case in the literature that documents Rhodotorula mucilaginosa fungemia associated with pleural tuberculosis.

  19. Vaccination with Recombinant Cryptococcus Proteins in Glucan Particles Protects Mice against Cryptococcosis in a Manner Dependent upon Mouse Strain and Cryptococcal Species

    Directory of Open Access Journals (Sweden)

    Charles A. Specht

    2017-11-01

    Full Text Available Development of a vaccine to protect against cryptococcosis is a priority given the enormous global burden of disease in at-risk individuals. Using glucan particles (GPs as a delivery system, we previously demonstrated that mice vaccinated with crude Cryptococcus-derived alkaline extracts were protected against lethal challenge with Cryptococcus neoformans and Cryptococcus gattii. The goal of the present study was to identify protective protein antigens that could be used in a subunit vaccine. Using biased and unbiased approaches, six candidate antigens (Cda1, Cda2, Cda3, Fpd1, MP88, and Sod1 were selected, recombinantly expressed in Escherichia coli, purified, and loaded into GPs. Three mouse strains (C57BL/6, BALB/c, and DR4 were then vaccinated with the antigen-laden GPs, following which they received a pulmonary challenge with virulent C. neoformans and C. gattii strains. Four candidate vaccines (GP-Cda1, GP-Cda2, GP-Cda3, and GP-Sod1 afforded a significant survival advantage in at least one mouse model; some vaccine combinations provided added protection over that seen with either antigen alone. Vaccine-mediated protection against C. neoformans did not necessarily predict protection against C. gattii. Vaccinated mice developed pulmonary inflammatory responses that effectively contained the infection; many surviving mice developed sterilizing immunity. Predicted T helper cell epitopes differed between mouse strains and in the degree to which they matched epitopes predicted in humans. Thus, we have discovered cryptococcal proteins that make promising candidate vaccine antigens. Protection varied depending on the mouse strain and cryptococcal species, suggesting that a successful human subunit vaccine will need to contain multiple antigens, including ones that are species specific.

  20. Serotipificación de aislamientos clínicos y del medio ambiente de Cryptococcus neoformans en Colombia

    Directory of Open Access Journals (Sweden)

    N. Ordóñez

    1994-09-01

    Full Text Available Se determinó l variedad y el serotipo de 192 aislamientos de Cryptococcus neoformans recuperados de 1972 a 1992; 163 de origen clínico y 29 del medio ambiente. En los aislamientos clínicos se determinaron las dos variedades de C. neoformans: la variedad neoformans en el 92,6% de los pacientes y la variedad gattii en el 7,4%. En el medio ambiente se encontró la variedad neoformans en el 100% de los aislamientos. El serotipo encontrado con más frecuencia fue el serotipo A, el cual se observó en el 92% de los aislamientos clínicos y en el 100% del medio ambiente, seguido por el serotipo B (6,8%; también, se determinaron aislamientos de los serotipos C (0,6% y D (0,6%. C. neoformans var. neoformans se recuperó en todos los años de los que se tuvieron aislamientos y C. neoformans var. gattii solamente se observó en aislamientos clínicos a partir de 1986. C. neoformans var. neoformans fue aislado durante los 12 meses del año y C. neoformans var. gattii se encontró distribuido en los meses defebrero, abril, junio, agosto, septiembre y diciembre. C. neoformans var. neoformans se observó en 109 (66,9% cepas de pacientes del sexo masculino y en 36 (22,1% del sexo femenino; C. neoformans var. gattii se observó en 5 (3% cepas de hombres y en 6 (3,7% mujeres.

  1. Immunotherapy of Cryptococcus infections.

    Science.gov (United States)

    Antachopoulos, C; Walsh, T J

    2012-02-01

    Despite appropriate antifungal treatment, the management of cryptococcal disease remains challenging, especially in immunocompromised patients, such as human immunodeficiency virus-infected individuals and solid organ transplant recipients. During the past two decades, our knowledge of host immune responses against Cryptococcus spp. has been greatly advanced, and the role of immunomodulation in augmenting the response to infection has been investigated. In particular, the role of 'protective' Th1 (tumour necrosis factor-α, interferon (IFN)-γ, interleukin (IL)-12, and IL-18) and Th17 (IL-23 and IL-17) and 'non-protective' Th2 (IL-4, IL-10, and IL-13) cytokines has been extensively studied in vitro and in animal models of cryptococcal infection. Immunomodulation with monoclonal antibodies against the capsular polysaccharide glucuronoxylomannan, glucosylceramides, melanin and β-glucan and, lately, with radioimmunotherapy has also yielded promising results in animal models. As a balance between sufficiently protective Th1 responses and excessive inflammation is important for optimal outcome, the effect of immunotherapy may range from beneficial to deleterious, depending on factors related to the host, the infecting organism, and the immunomodulatory regimen. Clinical evidence supporting immunomodulation in patients with cryptococcal infection remains too limited to allow firm recommendations. Limited human data suggest a role for IFN-γ. Identification of surrogate markers characterizing patients' immunological status could possibly suggest candidate patients for immunotherapy and the type of immunomodulation to be administered. © 2011 The Authors. Clinical Microbiology and Infection © 2011 European Society of Clinical Microbiology and Infectious Diseases.

  2. Seminational surveillance of fungemia in Denmark: notably high rates of fungemia and numbers of isolates with reduced azole susceptibility

    DEFF Research Database (Denmark)

    Arendrup, Maiken Cavling; Fuursted, Kurt; Gahrn-Hansen, Bente

    2005-01-01

    laboratory systems documented a continuous increase of candidemia cases since the early 1990s. For the 272 susceptibility-tested isolates, MICs of amphotericin B and caspofungin were within the limits expected for the species or genus. However, decreased azole susceptibility, defined as a fluconazole MIC...... of >8 microg/ml and/or itraconazole MIC of >0.125 microg/ml, was detected for 11 Candida isolates that were neither C. glabrata nor C. krusei. Including intrinsically resistant fungi, we detected decreased susceptibility to fluconazole and/or itraconazole in 87 (32%) current Danish bloodstream fungal...... isolates. We showed a continuous increase of fungemia in Denmark and an annual rate in 2003 to 2004 higher than in most other countries. The proportion of bloodstream fungal isolates with reduced susceptibility to fluconazole and/or itraconazole was also notably high....

  3. Cryptococcus neoformans hyperfilamentous strain is hypervirulent in a murine model of cryptococcal meningoencephalitis.

    Directory of Open Access Journals (Sweden)

    Marianna Feretzaki

    Full Text Available Cryptococcus neoformans is a human fungal pathogen that causes lethal infections of the lung and central nervous system in immunocompromised individuals. C. neoformans has a defined bipolar sexual life cycle with a and α mating types. During the sexual cycle, which can occur between cells of opposite mating types (bisexual reproduction or cells of one mating type (unisexual reproduction, a dimorphic transition from yeast to hyphal growth occurs. Hyphal development and meiosis generate abundant spores that, following inhalation, penetrate deep into the lung to enter the alveoli, germinate, and establish a pulmonary infection growing as budding yeast cells. Unisexual reproduction has been directly observed only in the Cryptococcus var. neoformans (serotype D lineage under laboratory conditions. However, hyphal development has been previously associated with reduced virulence and the serotype D lineage exhibits limited pathogenicity in the murine model. In this study we show that the serotype D hyperfilamentous strain XL280α is hypervirulent in an animal model. It can grow inside the lung of the host, establish a pulmonary infection, and then disseminate to the brain to cause cryptococcal meningoencephalitis. Surprisingly, this hyperfilamentous strain triggers an immune response polarized towards Th2-type immunity, which is usually observed in the highly virulent sibling species C. gattii, responsible for the Pacific Northwest outbreak. These studies provide a technological advance that will facilitate analysis of virulence genes and attributes in C. neoformans var. neoformans, and reveal the virulence potential of serotype D as broader and more dynamic than previously appreciated.

  4. Antifungal susceptibilities of Cryptococcus neoformans.

    Science.gov (United States)

    Archibald, Lennox K; Tuohy, Marion J; Wilson, Deborah A; Nwanyanwu, Okey; Kazembe, Peter N; Tansuphasawadikul, Somsit; Eampokalap, Boonchuay; Chaovavanich, Achara; Reller, L Barth; Jarvis, William R; Hall, Gerri S; Procop, Gary W

    2004-01-01

    Susceptibility profiles of medically important fungi in less-developed countries remain uncharacterized. We measured the MICs of amphotericin B, 5-flucytosine, fluconazole, itraconazole, and ketoconazole for Cryptococcus neoformans clinical isolates from Thailand, Malawi, and the United States and found no evidence of resistance or MIC profile differences among the countries.

  5. Estudio multicéntrico de fungemias por levaduras en la República Argentina Multicenter study of bloodstream infections due to yeasts in Argentina

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    L. Rodero

    2005-12-01

    Full Text Available La incidencia de candidemias aumentó aproximadamente en un 500% en hospitales de alta complejidad y se observó un cambio en la distribución de especies del género Candida, con un incremento de las levaduras no Candida albicans. Con el objeto de conocer la distribución de especies asociadas a fungemias por levaduras en Argentina y determinar su sensibilidad a los antifúngicos de uso convencional, se realizó un estudio multicéntrico durante el período abril 1999 a abril 2000. Participaron 36 instituciones del país. Se colectaron 265 aislamientos de levaduras provenientes de hemocultivos, que se identificaron utilizando pruebas morfológicas, fisiológicas y bioquímicas y la determinación de la concentración inhibitoria mínima se realizó en base al estándar del NCCLS. La distribución de especies fue: Candida albicans (40,75%, Candida parapsilosis (28,67%, Candida tropicalis (15,84%, Candida famata (3,77%, Cryptococcus neoformans (3,77%, Candida glabrata (2,64% y otras (4,53%. La mayoría de los aislamientos fueron sensibles a anfotericina B, fluconazol e itraconazol. La mortalidad asociada a las fungemias por levaduras estudiadas (n=265 fue del 30%, siendo más baja a lo descrito (33-54% y fue menor en los pacientes que recibieron tratamiento antifúngico (26,3%, que en los no tratados (47%.The incidence of candidemia has increased approximately 500% in high-complexity hospitals. A change in the spectrum of Candida infections due to species other than Candida albicans has also been detected. Between April 1999 and April 2000 a multicenter study was performed in order to determine the species distribution associated to candidemias in Argentina and the susceptibility profile of the isolates to the current antifungal drugs. Thirty six institutions have participated. All the 265 yeast strains isolated from blood cultures were identified by morphological, physiological, and biochemical tests. The antifungal susceptibility testing of

  6. Mesenteric cryptococcal granuloma in a dog caused by Cryptococcus neoformans var. grubii

    Directory of Open Access Journals (Sweden)

    Rodrigues-Hoffmann A

    2012-09-01

    Full Text Available Jennifer R Cook, Karen E Russell, Kristin B Eden, Aline Rodrigues-HoffmannDepartment of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX, USAAbstract: Although cryptococcosis is usually associated with respiratory and neurologic signs in domestic species (such as sneeze, cough, nasal discharge, seizures, ataxia, clinical manifestations of the disease may be more subtle and nonspecific. A 3-year-old male castrated Boxer dog presented with a history of chronic vomiting, diarrhea, weight loss, and lethargy. At no time had respiratory or neurologic signs been noted by the owners or the primary care veterinarian. Palpation of an abdominal mass revealed an atypical lesion location: a large (16 × 9 × 7 cm mass at the root of the mesentery. Diagnosis was achieved through cytology of this mass and a positive serologic Cryptococcus capsular antigen titer; polymerase chain reaction was utilized for speciation of the abdominal isolate as Cryptococcus neoformans variety grubii. The animal was euthanized due to poor prognosis. After necropsy and histopathologic analysis, the mesenteric mass and associated lymph nodes were identified as large fungal granulomas. This is a rare manifestation of cryptococcosis, involving several visceral organs, with no remaining evidence of the route of entry of the organism. As prompt diagnosis of mycotic illness is paramount to successful management, this case indicates that cryptococcal infection should be considered as a differential diagnosis in dogs with gastrointestinal signs and lymphadenopathy. The protean nature of cryptococcosis is discussed within the context of a brief review of emerging and unresolved issues in pathogenesis.Keywords: Cryptococcus gattii, granuloma, lymphadenitis

  7. Rhodotorula fungemia of an intensive care unit patient and review of published cases.

    Science.gov (United States)

    Spiliopoulou, Anastasia; Anastassiou, Evangelos D; Christofidou, Myrto

    2012-10-01

    Rhodotorula species are commensal yeasts that have emerged as a cause of life-threatening fungemia in severely immunocompromised patients. A case of Rhodotorula mucilaginosa fungemia in a 48-year-old woman that had undergone consecutive abdominal surgeries due to ovarian cancer and bowel necrosis while she was receiving fluconazole prophylaxis is presented. Several risk factors were identified such as presence of central venous catheters, solid organ neoplasm, abdominal surgery and administration of antibiotics. Identification was performed using commercial systems. The yeast was resistant to fluconazole, posaconazole and voriconazole and to echinocandins, whereas MIC to amphotericin B was 1.5 mg/L. Furthermore, published cases of Rhodotorula spp fungemia during the last decade are reviewed. In conclusion, Rhodotorula spp must be considered a potential pathogen in patients with immunosupression and central venous catheters. Correct identification is mandatory for appropriate management, as Rhodotorula spp are resistant to antifungal agents, such as fluconazole and echinocandins.

  8. [Cryptococcus neoformans chemotyping. Review of the literature. New epidemiologic data on cryptococcosis. Our experience with the use of C.G.B. media in the study of this yeast].

    Science.gov (United States)

    de Melo, N T; Lacaz, C da S; Charbel, C E; Pereira, A D; Heins-Vaccari, E M; França-Netto, A S; Machado, L dos R; Livramento, J A

    1993-01-01

    The purpose of this work was to collect the main information from the literature about the biotyping of Cryptococcus neoformans. The more up-to date research concerning the epidemiology of cryptococcosis comprising quite a few articles, mainly after the advent of AIDS, was also reviewed. The Cryptococcus neoformans varieties neoformans and gattii are well defined biochemically nowadays chiefly through the C.G.B. medium, according to Kwon-Chung et al. (1982). The isolation of C. neoformans var. gattii from flowers and leaves of Eucalyptus camaldulensis and Eucalyptus tereticornis, specially in Australia, through the works of Ellis & Pfeiffer (1990) and Pfeiffer & Ellis (1992) permitted very interesting epidemiological investigations on C. neoformans, a capsulated yeast by which Sanfelice, in Italy (1894; 1895) attracted attention of medical class. Busse, in 1894, described the first human case of cryptococcosis under the presentation of a bone lesion simulating sarcoma. In this paper, the Brazilian researchers focused on this subject were pointed out, followed by the Author's experience with the C.G.B. medium (L-canavanine, glycine and bromothymol blue) proposed by Kwon-Chung et al. (1982) with very good results. It was possible with such medium the study of 50 C.N.S. liquor samples, being 39 from AIDS patients (78%) and 11 from non-AIDS ones (22%). Thirty-seven out of the 39 HIV-positive patients (74%) were identified as C. neoformans var. gattii. From the negative HIV, 8 (16%) were classified as C. neoformans var. neoformans and 3 (6%) as C. neoformans var. gattii. We could not perform the serotyping of the above referred samples. It is evident anyway that in Brazil there exist both varieties gattii and neoformans, agents of neurocryptococcosis, including AIDS patients. The importance of neurocryptococcosis, mainly among AIDS patients, is stressed here, showing once more the value of C.G.B. medium in the typing of C. neoformans in its two varieties. Also, it is of

  9. Rhodotorula fungemia: Report of two cases in Sfax (Tunisia).

    Science.gov (United States)

    Guidara, R; Trabelsi, H; Neji, S; Cheikhrouhou, F; Sellami, H; Makni, F; Ayadi, A

    2016-06-01

    Rhodotorula is emerging as an important cause of nosocomial and opportunistic infections. We present two cases of Rhodotorula mucilaginosa fungemia diagnosed at our hospital during the last decade. The first case was of a term neonate who presented congenital heart disease (interventricular communication) and body dysmorphic disorder. He was admitted for respiratory failure and sepsis. The second case involved in a 33-year-old woman that had Hodgkinien lymphoma associated to tuberculosis. Identification was performed using commercial systems and confirmed by PCR sequencing of internal transcribed spacer, ITS1 and ITS2 regions of rDNA. Antifungal susceptibility tested by sensititre yeast revealed susceptibility to amphotericin B and resistance to fluconazole for the two strains. These cases emphasize the emerging importance of Rhodotorula sp. as a pathogen and it must be considered a potential pathogen in patients with immunosupression and with central venous catheters. Correct identification is mandatory for appropriate management, as Rhodotorula spp. are resistant to antifungal agents, such as fluconazole. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  10. Creatinine metabolism in Cryptococcus neoformans and Cryptococcus bacillisporus.

    OpenAIRE

    Polacheck, I; Kwon-Chung, K J

    1980-01-01

    The pathogenic species of Cryptococcus, C. neoformans and C. bacillisporus, utilized creatinine as a source of nitrogen but not of carbon. Chromatographic and autoradiographic studies suggest that creatinine metabolism in both species involves a single step resulting in the production of methylhydantoin and ammonia. The enzyme responsible for this step, creatinine deiminase, was produced by the cells only in the presence of creatinine in both species. The synthesis of creatinine deiminase was...

  11. Saccharomyces boulardii probiotic-associated fungemia: questioning the safety of this preventive probiotic's use.

    Science.gov (United States)

    Martin, Isabella W; Tonner, Rita; Trivedi, Julie; Miller, Heather; Lee, Richard; Liang, Xinglun; Rotello, Leo; Isenbergh, Elena; Anderson, Jennifer; Perl, Trish; Zhang, Sean X

    2017-03-01

    We report a case of fungemia in an immunocompetent patient after administration of probiotic containing Saccharomyces boulardii. We demonstrated the strain relatedness of the yeast from the probiotic capsule and the yeast causing fungal infection using genomic and proteomic typing methods. Our study questions the safety of this preventative biotherapy. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. FIRST REPORT ON Cryptococcus neoformans IN PIGEON EXCRETA FROM PUBLIC AND RESIDENTIAL LOCATIONS IN THE METROPOLITAN AREA OF CUIABÁ, STATE OF MATO GROSSO, BRAZIL

    Directory of Open Access Journals (Sweden)

    Doracilde Terumi Takahara

    2013-12-01

    Full Text Available SUMMARY Cryptococcosis is a severe systemic mycosis caused by two species of Cryptococcus that affect humans and animals: C. neoformans and C. gattii. Cosmopolitan and emergent, the mycosis results from the interaction between a susceptible host and the environment. The occurrence of C. neoformans was evaluated in 122 samples of dried pigeon excreta collected in 49 locations in the City of Cuiabá, State of Mato Grosso, Brazil, including public squares (n = 5, churches (n = 4, educational institutions (n = 3, health units (n = 8, open areas covered with asbestos (n = 4, residences (n = 23, factory (n = 1 and a prison (n = 1. Samples collected from July to December of 2010 were seeded on Niger seed agar (NSA. Dark brown colonies were identified by urease test, carbon source assimilation tests and canavanine-glycine-bromothymol blue medium. Polymerase chain reaction primer pairs specific for C. neoformans were also used for identification. Cryptococcus neoformans associated to pigeon excreta was isolated from eight (6.6% samples corresponding to six (12.2% locations. Cryptococcus neoformans was isolated from urban areas, predominantly in residences, constituting a risk of acquiring the disease by immunocompromised and immunocompetent individuals.

  13. First report on Cryptococcus neoformans in pigeon excreta from public and residential locations in the metropolitan area of Cuiabá, State of Mato Grosso, Brazil.

    Science.gov (United States)

    Takahara, Doracilde Terumi; Lazéra, Márcia dos Santos; Wanke, Bodo; Trilles, Luciana; Dutra, Valéria; Paula, Daphine Ariadne Jesus de; Nakazato, Luciano; Anzai, Mariana Caselli; Leite Júnior, Diniz Pereira; Paula, Claudete Rodrigues; Hahn, Rosane Christine

    2013-01-01

    Cryptococcosis is a severe systemic mycosis caused by two species of Cryptococcus that affect humans and animals: C. neoformans and C. gattii. Cosmopolitan and emergent, the mycosis results from the interaction between a susceptible host and the environment. The occurrence of C. neoformans was evaluated in 122 samples of dried pigeon excreta collected in 49 locations in the City of Cuiabá, State of Mato Grosso, Brazil, including public squares (n = 5), churches (n = 4), educational institutions (n = 3), health units (n = 8), open areas covered with asbestos (n = 4), residences (n = 23), factory (n = 1) and a prison (n = 1). Samples collected from July to December of 2010 were seeded on Niger seed agar (NSA). Dark brown colonies were identified by urease test, carbon source assimilation tests and canavanine-glycine-bromothymol blue medium. Polymerase chain reaction primer pairs specific for C. neoformans were also used for identification. Cryptococcus neoformans associated to pigeon excreta was isolated from eight (6.6%) samples corresponding to six (12.2%) locations. Cryptococcus neoformans was isolated from urban areas, predominantly in residences, constituting a risk of acquiring the disease by immunocompromised and immunocompetent individuals.

  14. Cryptococcus neoformans granuloma in the lung and spinal cord of a free-ranging cheetah (Acinonyx jubatus). A clinical report and literature review.

    Science.gov (United States)

    Millward, I R; Williams, M C

    2005-12-01

    A 6-year-old, male, wild-born, free-ranging cheetah (Acinonyx jubatus) was evaluated for acute onset of progressive lameness in the right hind limb. Survey radiographs were unrewarding and myelography indicated an intramedullary compressive mass at the L3-L4 region. A fine needle aspirate of the lesion indicated the presence of Cryptococcus organisms. Necropsy confirmed the presence of granulomas (cryptococcoma) in the lung and the spinal cord (meningomyelitis) caused by Cryptococcus neoformans var. gattii. Cryptococcus neoformans is a yeast-like organism that is a potential pathogen to many species. Initial infection is thought to be of respiratory origin and then it commonly disseminates systemically from the nasal cavity or lungs to the skin, eyes and central nervous system in particular. The cheetah tested negative for both feline leukaemia virus (FeLV) and feline immunodeficiency virus (FIV), as have all the previously reported cheetah cases. C. neoformans is a non-contagious, opportunistic organism and is the most common systemic mycoses in domestic cats and the cheetah.

  15. Cryptococcus neoformans granuloma in the lung and spinal cord of a free-ranging cheetah (Acinonyx jubatus. A clinical report and literature review : clinical communication

    Directory of Open Access Journals (Sweden)

    I.R. Millward

    2005-06-01

    Full Text Available A 6-year-old, male, wild-born, free-ranging cheetah (Acinonyx jubatus was evaluated for acute onset of progressive lameness in the right hind limb. Survey radiographs were unrewarding and myelography indicated an intramedullary compressive mass at the L3-L4 region. A fine needle aspirate of the lesion indicated the presence of Cryptococcus organisms. Necropsy confirmed the presence of granulomas (cryptococcoma in the lung and the spinal cord (meningomyelitis caused by Cryptococcus neoformans var. gattii. Cryptococcus neoformans is a yeast-like organism that is a potential pathogen to many species. Initial infection is thought to be of respiratory origin and then it commonly disseminates systemically from the nasal cavity or lungs to the skin, eyes and central nervous system in particular. The cheetah tested negative for both feline leukaemia virus (FeLV and feline immunodeficiency virus (FIV, as have all the previously reported cheetah cases. C. neoformans is a non-contagious, opportunistic organism and is the most common systemic mycoses in domestic cats and the cheetah.

  16. Quimiotipagem do Cryptococcus neoformans. Revisão da literatura. Novos dados epidemiológicos sobre a criptococose. Nossa experiência com o emprego do meio de C.G.B. no estudo daquela levedura Biotyping of Cryptococcus neoformans. Review of the literature. New epidemiologic informations about cryptococcosis. Our experience with the utilization of C.G.B medium in this yeast

    Directory of Open Access Journals (Sweden)

    Natalina Takahashi de Melo

    1993-10-01

    Full Text Available O presente trabalho, além da revisão da literatura sobre quimiotipagem do C. neoformans, com novos dados sobre a epidemiologia da criptococose, teve por finalidade principal a caracterização das duas variedades desta levedura em pacientes com neurocriptococose, HIV + e HIV -. As variedades neoformans e gattii estão hoje bem definidas bioquimicamente, com o emprego do meio C.G.B., proposto por KWON-CHUNG et al. (1982 24. O isolamento do C. neoformans var. gattii das flores e folhas do Eucalyptus camaldulensis e do Eucalyptus tereticornis, na Austrália, através dos trabalhos de ELLIS & PFEIFFER (199016 e PFEIFFER & ELLIS (199241, possibilitou investigações epidemiológicas das mais interessantes sobre este microrganismo, levedura capsulada a qual SANFELICE50, 51, na Itália, em 1894 e 1895 despertou a atenção do meio médico. BUSSE8, em 1894, descrevia o primeiro caso de criptococose humana sob a forma de lesão óssea, simulando sarcoma. As pesquisas nacionais sobre o assunto em foco foram destacadas, seguindo-se a experiência dos Autores com o meio de C.G.B. (L - canavanina, glicina e azul de bromotimol. Foi possível, através deste meio o estudo de 50 amostras de líquor, sendo 39 procedentes de aidéticos (78% e 11 de não aidéticos (22%. De pacientes HIV+, 37 (74% foram identificados como C. neoformans var. neoformans e 2 (4% como C. neoformans var. gattii. Dos HIV- 8 ( 16% foram classificados como C. neoformans var. neoformans e 3 (6% como C. neoformans var. gattii. Através deste trabalho, evidencia-se a importância da neurocriptococose, principalmente entre os aidéticos, demonstrando-se mais uma vez o interesse do meio CGB na quimiotipagem do C. neoformans em suas duas variedades, ganhando em importância a demonstração de que duas espécies de eucalipto podem funcionar como "árvores-hospedeiras" para o Cryptococcus neoformans var. gattii.The purpose of this work was to collect the main information from the literature

  17. Molecular characterization of Cryptococcus neoformans isolated from the environment in Beijing, China.

    Science.gov (United States)

    Dou, Hongtao; Wang, Huizhu; Xie, Shaowei; Chen, Xinxin; Xu, Zhipeng; Xu, Yingchun

    2017-10-01

    The molecular type of environmental Cryptococcus neoformans in Beijing was not clear. Our study aims to reveal the molecular characterization of C. neoformans complex from environment in Beijing, China. A total of 435 samples of pigeon droppings from 11 different homes in Beijing were collected from August to November in 2015. Pigeon droppings were inoculated onto caffeic acid cornmeal agar (CACA) to screen C. neoformans complex. Bruker Biotyper matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was performed for species identification. Serotype and mating type was determined by specific primers. Restriction fragment length polymorphisms of URA5 (URA5-RFLP) were applied to genotype. Multi-locus sequence typing (MLST) was done for further identification and sequence type (ST) determination. Altogether, 81 isolates of C. neoformans AFLP1/VNI were recognized from 435 pigeon droppings in this study. The positive rate for C. neoformans AFLP1/VNI from pigeon droppings in different homes varied from 5.0% to 52.6%, the average was 20.2%. All of these cryptococcal strains were serotype A, MATα. They were genotyped as VNI by URA5-RFLP and were confirmed by MLST. No other molecular types of C. neoformans and Cryptococcus gattii isolates were isolated. Their STs were identified as ST 31 (n = 54, 66.7%), followed by ST 53 (n = 10), ST 191 (n = 8), ST 5 (n = 5), ST 57 (n = 3), and ST 38 (n = 1). We concluded that not only clinical but also environmental isolates of C. neoformans need to be investigated more deeply and more extensively. The virulence difference between ST 5 and ST 31 need to be explored in the future. © The Author 2017. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Molecular characterization and antifungal susceptibility of Cryptococcus neoformans strains collected from a single institution in Lima, Peru.

    Science.gov (United States)

    Bejar, Vilma; Tello, Mercedes; García, Ruth; Guevara, José M; Gonzales, Sofia; Vergaray, German; Valencia, Esther; Abanto, Enma; Ortega-Loayza, Alex G; Hagen, Ferry; Gutierrez, Ericson L

    2015-01-01

    Cryptococcosis is a fungal infection with a worldwide distribution, mainly caused by Cryptococcus neoformans and Cryptococcus gattii. To molecularly characterize the mating-types, serotypes, genotypes and antifungal susceptibility profiles of a set of retrospectively isolated C. neoformans strains from Lima, Peru. A set of 32 Cryptococcus spp. strains from the Institute of Tropical Medicine of the National University of San Marcos, Lima, Peru, were included in this retrospective study. Twenty-four strains were isolated from patients, while the remaining 8 were isolated from the environment. Using conventional PCR, 27 (84.4%) of the isolates were identified as C. neoformans var. grubii mating-type alpha and serotype A. Using the AFLP fingerprinting, it was shown that 16 (50%) of the C. neoformans strains were genotype AFLP1, 13 (40.6%) were genotype AFLP1B, 2 (6.3%) were genotype AFLP2, and 1 (3.1%) was found to be a hybrid between both C. neoformans varieties (genotype AFLP3). The antifungal susceptibility profiles for amphotericin B, fluconazole and voriconazole showed that all the 32 C. neoformans are sensitive to these antifungal compounds. In this study we observed that C. neoformans var. grubii (AFLP1 and AFLP1B) and C. neoformans var. neoformans (AFLP2) were the only cryptococcal varieties involved. All strains were found to be sensitive to the antifungals tested, results that are consistent with those found in the international literature. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  19. Malassezia pachydermatis fungemia in a preterm neonate resistant to fluconazole and flucytosine

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    Noura Al-Sweih

    2014-07-01

    Full Text Available A case of Malassezia pachydermatis fungemia in a preterm neonate is described. The isolate was identified by rDNA sequencing and was resistant to fluconazole and flucytosine. Since M. pachydermatis does not require lipid supplementation for growth, it can be misidentified as a Candida species. The report highlights M. pachydermatis as a cause of late onset sepsis in preterm neonates and emphasizes the need for prior antifungal susceptibility testing.

  20. Comparison of the lysis-centrifugation and agitated biphasic blood culture systems for detection of fungemia.

    Science.gov (United States)

    Murray, P R

    1991-01-01

    Although the detection of fungemia has been improved by the use of vented or biphasic blood culture bottles, the best recovery and earliest detection have been reported in the Isolator lysis-centrifugation system. It was recently demonstrated that improved detection of both bacteria and fungi was accomplished by mechanically agitating blood culture bottles for the first 24 h of incubation. In this study the detection of fungemia by use of the Isolator system was compared with that of an agitated biphasic system. A total of 182 fungi were isolated from blood specimens inoculated into both culture systems. No difference in the overall recovery of fungi or individual species of yeasts was observed between the two systems. However, all seven isolates of Histoplasma capsulatum were recovered in the Isolator system only. The time required to detect fungemia with each of the two systems was also compared. No statistically significant difference was observed. From the data collected during this 18-month study, it can be concluded that the overall recovery and time of detection of yeasts are equivalent in the lysis-centrifugation system and the agitated biphasic blood culture system. The lysis-centrifugation system is still superior for the detection of filamentous fungi such as H. capsulatum. PMID:1993772

  1. Caracterización fenotípica de aislamientos ambientales de Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Sandra Huérfano

    2003-09-01

    Full Text Available La criptococosis es causada por las tres variedades de Cryptococcus neoformans, las cuales presentan diferencias fisiológicas y de virulencia, algunas de las cuales se han estudiado para reconocer aspectos de su biología. Este trabajo evaluó las características fenotípicas de aislamientos ambientales de las variedades grubii y gattii, con el fin de establecer diferencias asociadas con el ciclo de vida y la virulencia. Se estudiaron 28 aislamientos serotipo A y 31 serotipo C. Se evaluaron la morfología macroscópica y microscópica de blastoconidias cultivadas en agar Sabouraud y suelos, el crecimiento a 37 °C, la producción de 22 enzimas extracelulares, la frecuencia del fenómeno de fructificación haploide, la pareja sexual, el patrón de sensibilidad a toxinas asesinas (killer y la virulencia en ratones Balb/c. No se observaron diferencias entre las dos variedades en la morfología macroscópica, la microscópica, ni en el crecimiento a 37 °C (p>0,05; sin embargo, se observó disminución del tamaño celular y capsular de blastoconidias cultivadas en suelo comparado con el tamaño en Sabouraud (p

  2. Cryptococcus

    Indian Academy of Sciences (India)

    Vikas Yadav

    The Molecular Mycology Laboratory. CSIR, DBT, SERB,. CEFIPRA (Indo-French), IJSPS (Indo-Japan). JNCASR (intramural). TATA innovation Fellowship. Rahul Siddharthan. DD Dubey. German Larriba. Joseph Heitman.

  3. Incidence and Clinical Predictors of Ocular Candidiasis in Patients with Candida Fungemia

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    Ayesha Khalid

    2014-01-01

    Full Text Available Purpose. The aim of this study is to determine the incidence and the predictors of ocular candidiasis among patient with Candida fungemia. Methods. We retrospectively reviewed the charts of all patients diagnosed with candidemia at the University of Kansas Medical Center during February 2000–March 2010. Data regarding patients’ demographics, clinical characteristics, laboratory results, and ophthalmology examination findings were collected. Results. A total of 283 patients with candidemia were enrolled. The mean age (± standard deviation was 55 ± 18 years; 66% were male. The most commonly isolated Candida species were C. albicans (54%, C. parapsilosis (20%, C. glabrata (13%, and C. tropicalis (8%. Only 144 (51% patients were evaluated by ophthalmology; however, the proportion of patients who were formally evaluated by an ophthalmologist increased during the study period (9%in 2000 up to 73%in 2010; P<0.0001. Evidence of ocular candidiasis was present in 18 (12.5% patients. Visual symptoms were reported by 5 of 18 (28% patients. In multivariable analysis, no predictors of ocular candidiasis were identified. Conclusions. The incidence of ocular candidiasis among patients with fungemia remains elevated. Most patients are asymptomatic and therefore all patients with candidemia should undergo fundoscopic examination to rule out ocular involvement.

  4. A dedicated fungal culture medium is useful in the diagnosis of fungemia: a retrospective cross-sectional study.

    Science.gov (United States)

    Zheng, Shuwei; Ng, Tong Yong; Li, Huihua; Tan, Ai Ling; Tan, Thuan Tong; Tan, Ban Hock

    2016-01-01

    Mortality for candidemia ranges from 15% to 35%. Current guidelines recommend inoculating blood into three aerobic and three anaerobic blood culture bottles when candidemia is suspected, without mention of a fungal blood culture bottle. To determine the value of the BACTEC Myco/F Lytic blood culture media in the diagnosis of fungemia. A two-year retrospective cross-sectional study was performed for patients who had fungemia with submitted BACTEC Plus Aerobic/F (Aer), BACTEC Plus Anaerobic/F (Anaer) or Myco/F Lytic (Myco) blood culture bottles. The detection rate of fungemia was 77.4% in 93 patients with contemporaneously submitted blood culture bottles when limited to only Aer/Anaer culture results. The detection rate improved significantly with the addition of the Myco culture bottle results (pculture bottle submissions were less useful for patients with appropriate anti-fungal therapy administered within 48 hours [OR = 0.18, 95% CI = (0.06, 0.49), p = 0.001] and those with fungal growth detected within 48 hours [OR = 0.33, 95% CI = (0.12, 0.89), p = 0.001]. Among a subset of patients with concordant blood culture results, those with Myco culture bottles submission allowed earlier fungal detection and speciation by at least one day in 27.5% and 25.0% of the cases respectively. Our study highlights the importance of a dedicated fungal blood culture when fungemia is clinically suspected. Nearly a quarter of fungemias may be missed if a fungal blood culture is not performed.

  5. Cryptococcus neoformans carried by Odontomachus bauri ants

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    Mariana Santos de Jesus

    2012-06-01

    Full Text Available Cryptococcus neoformans is the most common causative agent of cryptococcosis worldwide. Although this fungus has been isolated from a variety of organic substrates, several studies suggest that hollow trees constitute an important natural niche for C. neoformans. A previously surveyed hollow of a living pink shower tree (Cassia grandis positive for C. neoformans in the city of Rio de Janeiro, Brazil, was chosen for further investigation. Odontomachus bauri ants (trap-jaw ants found inside the hollow were collected for evaluation as possible carriers of Cryptococcus spp. Two out of 10 ants were found to carry phenoloxidase-positive colonies identified as C. neoformans molecular types VNI and VNII. The ants may have acted as a mechanical vector of C. neoformans and possibly contributed to the dispersal of the fungi from one substrate to another. To the best of our knowledge, this is the first report on the association of C. neoformans with ants of the genus Odontomachus.

  6. Infecciones fúngicas emergentes: fungemia por Sporobolomyces salmonicolor. Reporte de un caso Emerging fungal infections: Sporobolomyces salmonicolor fungemia. Case Report

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    Adriana Paola Franco Rodríguez

    2010-12-01

    Full Text Available En los últimos años, la emergencia de infecciones por hongos se ha dado en parte por el aumento en las infecciones por microrganismos que normalmente no son patógenos. El siguiente reporte describe la fungemia y meningoencefalitis por Sporobolomyces salmonicolor, una levadura frecuente en el medio ambiente y las plantas, en un paciente sin factores de riesgo conocidos, con desenlace fatal a pesar del manejo con antifúngicos. Este caso nos recuerda que dentro de las infecciones micóticas emergentes se encuentra el S. salmonicolor y se debe tener en cuenta dentro del diagnóstico diferencial de levaduras en sangre, como causa infrecuente, potencialmente mortal.The emergence of pathogens, mainly fungi, in recent years has been partly due to overappearance of organisms that normally are not pathogenic in humans. The following report describes Sporobolomyces salmonicolor fungaemia and proven meningoencephalitis in a patient not showing risk factors who suffers a fatal outcome despite management with antifungal agents. The said yeast lives mainly in tree environments and plants. This case reminds us that there is S. salmonicolor within the emerging fungal infections, and this fact should be taken into account in the differential diagnosis of yeasts in blood as a rare and lifethreatening cause.

  7. Fungemia due to Rhodotorula mucilaginosa after allogeneic hematopoietic stem cell transplantation.

    Science.gov (United States)

    Mori, T; Nakamura, Y; Kato, J; Sugita, K; Murata, M; Kamei, K; Okamoto, S

    2012-02-01

    Rhodotorula species have been increasingly recognized as emerging pathogens, particularly in immunocompromised patients. We herein report on a patient with myelodysplastic syndrome who developed fungemia due to Rhodotorula mucilaginosa after allogeneic hematopoietic stem cell transplantation (HSCT) from an unrelated donor. He developed severe acute graft-versus-host disease requiring high-dose steroids, and had serially been administered fluconazole and micafungin for the prophylaxis of fungal infection. Although several cases of Rhodotorula infection after HSCT have been reported, all of them were recipients of autologous HSCT, not allogeneic HSCT. A review of all the reported cases of Rhodotorula infection after HSCT revealed that all patients had received fluconazole or echinocandins before the onset of infection. The findings suggest that Rhodotorula species could be causative yeasts, particularly in patients receiving fluconazole or echinocandins, both of which are inactive against the species. © 2011 John Wiley & Sons A/S.

  8. Successful Treatment of Candida parapsilosis Fungemia in Two Preterms with Voriconazole

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    Emel Altuncu

    2015-01-01

    Full Text Available Herein, we report two preterms with invasive candidiasis refractory to liposomal amphotericin B (AMB treatment in spite of low MIC levels (MIC: 0.5 mcg/mL. Both of the patients’ blood cultures were persistently positive for C. parapsilosis despite high therapeutic doses (AMB: 7 mg/kg per day. After starting voriconazole blood cultures became negative and both of the patients were treated successfully without any side effects. In conclusion, although it is not a standard treatment in neonatal patients, our limited experience with these patients suggests that voriconazole appears to be a safe antifungal agent to be used in critically ill preterm infants with persistent fungemia despite AMB treatment.

  9. A prospective descriptive study of cryptococcal meningitis in HIV uninfected patients in Vietnam - high prevalence of Cryptococcus neoformans var grubii in the absence of underlying disease

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    Baker Stephen

    2010-07-01

    Full Text Available Abstract Background Most cases of cryptococcal meningitis occur in patients with HIV infection: the course and outcome of disease in the apparently immunocompetent is much more poorly understood. We describe a cohort of HIV uninfected Vietnamese patients with cryptococcal meningitis in whom underlying disease is uncommon, and relate presenting features of patients and the characteristics of the infecting species to outcome. Methods A prospective descriptive study of HIV negative patients with cryptococcal meningitis based at the Hospital for Tropical Diseases, Ho Chi Minh City. All patients had comprehensive clinical assessment at baseline, were cared for by a dedicated study team, and were followed up for 2 years. Clinical presentation was compared by infecting isolate and outcome. Results 57 patients were studied. Cryptococcus neoformans var grubii molecular type VN1 caused 70% of infections; C. gattii accounted for the rest. Most patients did not have underlying disease (81%, and the rate of underlying disease did not differ by infecting species. 11 patients died while in-patients (19.3%. Independent predictors of death were age ≥ 60 years and a history of convulsions (odds ratios and 95% confidence intervals 8.7 (1 - 76, and 16.1 (1.6 - 161 respectively. Residual visual impairment was common, affecting 25 of 46 survivors (54.3%. Infecting species did not influence clinical phenotype or outcome. The minimum inhibitory concentrations of flucytosine and amphotericin B were significantly higher for C. neoformans var grubii compared with C. gattii (p Conclusion In HIV uninfected individuals in Vietnam, cryptococcal meningitis occurs predominantly in people with no clear predisposing factor and is most commonly due to C. neoformans var grubii. The rates of mortality and visual loss are high and independent of infecting species. There are detectable differences in susceptibility to commonly used antifungal drugs between species, but the clinical

  10. Interspecies differences in the enantioselectivity of epoxide hydrolases in Cryptococcus laurentii (Kufferath) C.E. Skinner and Cryptococcus podzolicus (Bab'jeva & Reshetova) Golubev

    CSIR Research Space (South Africa)

    Botes, AL

    2005-01-01

    Full Text Available Isolates representing Cryptococcus laurentii and Cryptococcus podzolicus, originating from soil of a heath land indigenous to South Africa, were screened for the presence of enantioselective epoxide hydrolases for 2, 2-disubstituted epoxides...

  11. Isolation of 2-deoxy-scyllo-inosose (DOI)-assimilating yeasts and cloning and characterization of the DOI reductase gene of Cryptococcus podzolicus ND1.

    Science.gov (United States)

    Ara, Satoshi; Yamazaki, Harutake; Takaku, Hiroaki

    2018-04-01

    2-Deoxy-scyllo-inosose (DOI) is the first intermediate in the 2-deoxystreptamine-containing aminoglycoside antibiotic biosynthesis pathway and has a six-membered carbocycle structure. DOI is a valuable material because it is easily converted to aromatic compounds and carbasugar derivatives. In this study, we isolated yeast strains capable of assimilating DOI as a carbon source. One of the strains, Cryptococcus podzolicus ND1, mainly converted DOI to scyllo-quercitol and (-)-vibo-quercitol, which is a valuable compound used as an antihypoglycemia agent and as a heat storage material. An NADH-dependent DOI reductase coding gene, DOIR, from C. podzolicus ND1 was cloned and successfully overexpressed in Escherichia coli. The purified protein catalyzed the irreversible reduction of DOI with NADH and converted DOI into (-)-vibo-quercitol. The enzyme had an optimal pH of 8.5 and optimal temperature of 35°C, respectively. The k cat of this enzyme was 9.98 s -1 , and the K m values for DOI and NADH were 4.38 and 0.24 mM, respectively. The enzyme showed a strong preference for NADH and showed no activity with NADPH. Multiple-alignment analysis of DOI reductase revealed that it belongs to the GFO_IDH_MocA protein family and is an inositol dehydrogenase homolog in other fungi, such as Cryptococcus gattii, and bacteria, such as Bacillus subtilis. This is the first identification of a DOI-assimilating yeast and a gene involved in DOI metabolism in fungi. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  12. Serotype, mating type and ploidy of Cryptococcus neoformans strains isolated from patients in Brazil Sorotipos, "mating type" e ploidia de amostras de C. neoformans isoladas de pacientes no Brasil

    Directory of Open Access Journals (Sweden)

    Misako OHKUSU

    2002-12-01

    Full Text Available Serotype, mating type and ploidy of 84 strains of Cryptococcus neoformans isolated from 61 AIDS and 23 non-AIDS patients admitted in a tertiary teaching hospital in São Paulo, Brazil were examined. Among 61 strains isolated from AIDS patients, 60 strains were var. grubii (serotype A. Only one strain was var. gattii (serotype B. No var. neoformans (serotype D was found. Among 23 strains isolated from non-AIDS patients, 15 were var. grubii (serotype A and the remaining 8 were var. gattii, all of which were serotype B. Seventy-three of the 75 serotype A strains were the heterothallic alpha type (MATalpha and the remaining 2 were untypable (asexual. Most of the MATalpha strains (69/73 were haploid and the remaining 4 strains were diploid. Similarly, both of the 2 asexual strains among the 75 serotype A strains were haploid. There were no alpha-mating type (MATalpha strains among the 84 isolates. All of the 8 var. gattii strains were serotype B and haploid. Among a total of 84 strains tested, neither serotype AD nor serotype D were found. Neither triploid nor tetraploid were found. These results suggest that the serological, sexual and ploidy characteristics in C. neoformans strains isolated from AIDS patients in São Paulo were rather simple, whereas strains isolated from non-AIDS patients presented serotype A and B with predominance of serotype A.Foram estudados os sorotipos, "mating type" e ploidia de 84 amostras de C. neoformans isoladas de 61 pacientes com AIDS e 23 não-AIDS em São Paulo. Das amostras isoladas de pacientes com AIDS, 60 foram identificadas como var. grubii (sorotipo A e 1 como var. gattii (sorotipo B. Não houve isolamento do sorotipo D. Entre as amostras isoladas, de pacientes não-AIDS, 15 foram de var. grubii (sorotipo A e as 8 restantes de var. gattii, todos do sorotipo B. Setenta e três dos 75 sorotipos A foram identificadas como cepas heterotálicas do fenótipo alfa (MATalfa e as 2 remanescentes n

  13. Sterylglucoside catabolism in Cryptococcus neoformans with endoglycoceramidase-related protein 2 (EGCrP2), the first steryl-β-glucosidase identified in fungi.

    Science.gov (United States)

    Watanabe, Takashi; Ito, Tomoharu; Goda, Hatsumi M; Ishibashi, Yohei; Miyamoto, Tomofumi; Ikeda, Kazutaka; Taguchi, Ryo; Okino, Nozomu; Ito, Makoto

    2015-01-09

    Cryptococcosis is an infectious disease caused by pathogenic fungi, such as Cryptococcus neoformans and Cryptococcus gattii. The ceramide structure (methyl-d18:2/h18:0) of C. neoformans glucosylceramide (GlcCer) is characteristic and strongly related to its pathogenicity. We recently identified endoglycoceramidase-related protein 1 (EGCrP1) as a glucocerebrosidase in C. neoformans and showed that it was involved in the quality control of GlcCer by eliminating immature GlcCer during the synthesis of GlcCer (Ishibashi, Y., Ikeda, K., Sakaguchi, K., Okino, N., Taguchi, R., and Ito, M. (2012) Quality control of fungus-specific glucosylceramide in Cryptococcus neoformans by endoglycoceramidase-related protein 1 (EGCrP1). J. Biol. Chem. 287, 368-381). We herein identified and characterized EGCrP2, a homologue of EGCrP1, as the enzyme responsible for sterylglucoside catabolism in C. neoformans. In contrast to EGCrP1, which is specific to GlcCer, EGCrP2 hydrolyzed various β-glucosides, including GlcCer, cholesteryl-β-glucoside, ergosteryl-β-glucoside, sitosteryl-β-glucoside, and para-nitrophenyl-β-glucoside, but not α-glucosides or β-galactosides, under acidic conditions. Disruption of the EGCrP2 gene (egcrp2) resulted in the accumulation of a glycolipid, the structure of which was determined following purification to ergosteryl-3β-glucoside, a major sterylglucoside in fungi, by mass spectrometric and two-dimensional nuclear magnetic resonance analyses. This glycolipid accumulated in vacuoles and EGCrP2 was detected in vacuole-enriched fraction. These results indicated that EGCrP2 was involved in the catabolism of ergosteryl-β-glucoside in the vacuoles of C. neoformans. Distinct growth arrest, a dysfunction in cell budding, and an abnormal vacuole morphology were detected in the egcrp2-disrupted mutants, suggesting that EGCrP2 may be a promising target for anti-cryptococcal drugs. EGCrP2, classified into glycohydrolase family 5, is the first steryl

  14. Pseudomonas aeruginosa inhibits the growth of Cryptococcus species.

    Science.gov (United States)

    Rella, Antonella; Yang, Mo Wei; Gruber, Jordon; Montagna, Maria Teresa; Luberto, Chiara; Zhang, Yong-Mei; Del Poeta, Maurizio

    2012-06-01

    Pseudomonas aeruginosa is a ubiquitous and opportunistic bacterium that inhibits the growth of different microorganisms, including Gram-positive bacteria and fungi such as Candida spp. and Aspergillus fumigatus. In this study, we investigated the interaction between P. aeruginosa and Cryptococcus spp. We found that P. aeruginosa PA14 and, to a lesser extent, PAO1 significantly inhibited the growth of Cryptococcus spp. The inhibition of growth was observed on solid medium by the visualization of a zone of inhibition of yeast growth and in liquid culture by viable cell counting. Interestingly, such inhibition was only observed when P. aeruginosa and Cryptococcus were co-cultured. Minimal inhibition was observed when cell-cell contact was prevented using a separation membrane, suggesting that cell contact is required for inhibition. Using mutant strains of Pseudomonas quinoline signaling, we showed that P. aeruginosa inhibited the growth of Cryptococcus spp. by producing antifungal molecules pyocyanin, a redox-active phenazine, and 2-heptyl-3,4-dihydroxyquinoline (PQS), an extracellular quorum-sensing signal. Because both P. aeruginosa and Cryptococcus neoformans are commonly found in lung infections of immunocompromised patients, this study may have important implication for the interaction of these microbes in both an ecological and a clinical point of view.

  15. Neurocriptococose por Cryptococcus neoformans não capsulado

    OpenAIRE

    Lacaz, C.S.; Heins-Vaccari, Elisabeth M.; Melo, Natalina T.; Moreno-Carvalho, O.A.; Sampaio, M.L.S.; Nogueira, L.S.; Badaró, R.J.S.; Livramento, J.A.

    1993-01-01

    Os autores registram um caso de neurocriptococose em paciente HIV-negativo, por Cryptococcus neoformans acapsulado ou deficiente em cápsula. O quadro neurológico era de meningoencefalite subaguda, compatível ao diagnóstico de neurotuberculose, pelo exame do líquido cefalorraqueano (LCR), Estruturas leveduriformes foram encontradas no interior de macrófagos, ao exame citomorfológico do LCR. Cultivo do sedimento do LCR revelou a presença de Cryptococcus neoformans não capsulado (identificação b...

  16. Cryptococcus laurentii fungaemia in a cervical cancer patient

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    Rejane Pereira Neves

    2015-11-01

    Full Text Available Infections caused by emerging Cryptococcus non-neoformans species are being reported with increasingly frequency. Here, we present a case of fungaemia by Cryptococcus laurentii in a woman receiving aggressive immunosuppressive therapy for cervical neoplasia. Three venous blood samples were aseptically collected on consecutive days and C. laurentii was isolated and identified through phenotypic and molecular methods. After central venous catheter removal and appropriate antifungal therapy, the patient showed significant improvement and blood culture became negative. Thus, patients following immunosuppressive therapies and using invasive medical devices are at risk of C. laurentii blood infections.

  17. Cryptococcus neoformans modulates extracellular killing by neutrophils

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    Asfia eQureshi

    2011-09-01

    Full Text Available We recently established a key role for host sphingomyelin synthase (SMS in the regulation of the killing activity of neutrophils against Cryptococcus neoformans. In this work, we studied the effect of C. neoformans on the killing activity of neutrophils and whether SMS would still be a player against C. neoformans in immunocompromised mice lacking T and NK cells (Tgε26 mice. To this end, we analyzed whether C. neoformans would have any effect on neutrophil survival and killing in vitro and in vivo. We show that unlike C. albicans, neither the presence nor the capsule size of C. neoformans cells have any effect on neutrophil viability. Interestingly, melanized C. neoformans cells totally abrogated the killing activity of neutrophils. Next, we monitored how exposure of neutrophils to C. neoformans cells would interfere with any further killing activity of the medium and found that pre-incubation with live but not heat-killed fungal cells significantly inhibits further killing activity of the medium. We next studied whether activation of SMS at the site of C. neoformans infection is dependent on T and NK cells. Using matrix-assisted laser desorption-ionization (MALDI tissue imaging in infected lung we found that similarly to previous observations in the isogenic wild type CBA/J mice, SM 16:0 levels are significantly elevated at the site of infection in mice lacking T and NK cells but only at early time points. This study highlights that C. neoformans may negatively regulate the killing activity of neutrophils and that SMS activation in neutrophils appears to be partially independent of T and/or NK cells.

  18. A novel xylosylphosphotransferase activity discovered in Cryptococcus neoformans

    DEFF Research Database (Denmark)

    Reilly, Morgann C; Levery, Steven B; Castle, Sherry A

    2009-01-01

    Cryptococcus neoformans is a fungal pathogen that causes serious disease in immunocompromised individuals. The organism produces a distinctive polysaccharide capsule that is necessary for its virulence, a predominantly polysaccharide cell wall, and a variety of protein- and lipid-linked glycans...

  19. Peripheral blood and bone marrow cells cultivated in Novy-Mcneal-Nicolle medium for visceral leishmaniosis diagnosis revealed Rhodotorula fungemia in an AIDS patient with lymphoma.

    Science.gov (United States)

    Paugam, Andre; Lebuisson, Agathe; Lortholary, Olivier; Baixench, Marie-Therese; Lanternier, Fanny

    2013-01-01

    Rhodotorula is a ubiquitous yeast that can infect immunocompromised patients. Here, we present the case of a 45-year-old patient with AIDS who developed a Rhodotorula mucilaginosa fungemia. The patient had a past history of visceral leishmaniasis (VL) and was hospitalised to receive chemotherapy for a B-cell lymphoma of the sinonasal cavities. The patient had no fever and no signs of VL. A systematic research for Leishmania by blood and bone marrow cultures was made and he received liposomal amphtotericin B (3 mg/kg in a single dose) to prevent a VL relapse. Rhodotorula fungemia was accidentally detected after 17 days of blood culture using a specific medium for leishmaniasis diagnosis. This long culture incubation time was probably facilitated by amphotericin B treatment. Rhodotorula is an emerging pathogen that may escape detection due its slow growth in culture.

  20. Catheter-related Saccharomyces cerevisiae Fungemia Following Saccharomyces boulardii Probiotic Treatment: In a child in intensive care unit and review of the literature

    Directory of Open Access Journals (Sweden)

    Serkan Atıcı

    2017-03-01

    Full Text Available Although Saccharomyces boulardii is usually a non-pathogenic fungus, in rare occasions it can cause invasive infection in children. We present the case of an 8-year-old patient in pediatric surgical intensive care unit who developed S. cerevisiae fungemia following probiotic treatment containing S. boulardii. Caspofungin was not effective in this case and he was treated with amphotericin B. We want to emphasize that physicians should be careful about probiotic usage in critically ill patients.

  1. Catheter-related Saccharomyces cerevisiae Fungemia Following Saccharomyces boulardii Probiotic Treatment: In a child in intensive care unit and review of the literature.

    Science.gov (United States)

    Atıcı, Serkan; Soysal, Ahmet; Karadeniz Cerit, Kıvılcım; Yılmaz, Şerife; Aksu, Burak; Kıyan, Gürsu; Bakır, Mustafa

    2017-03-01

    Although Saccharomyces boulardii is usually a non-pathogenic fungus, in rare occasions it can cause invasive infection in children. We present the case of an 8-year-old patient in pediatric surgical intensive care unit who developed S. cerevisiae fungemia following probiotic treatment containing S. boulardii . Caspofungin was not effective in this case and he was treated with amphotericin B. We want to emphasize that physicians should be careful about probiotic usage in critically ill patients.

  2. DNA probe for strain typing of Cryptococcus neoformans.

    OpenAIRE

    Varma, A; Kwon-Chung, K J

    1992-01-01

    A 7-kb linear plasmid, harbored by a URA5 transformant, hybridized to all the chromosomes of Cryptococcus neoformans separated by contour-clamped homogeneous electric field electrophoresis. Its linear maintenance was determined to have been facilitated by the presence of telomere-like sequences at its free ends. Hybridization of this plasmid to AccI-digested genomic DNAs of 26 C. neoformans strains generated 21 unique DNA fingerprints. The DNA fingerprints of isolates within the same serotype...

  3. Identification of Cryptococcus neoformans isolates using Staib agar without creatinine

    OpenAIRE

    Nardelli, Vanessa; Pérez, Celina; Mata-Essayag, Sofía; Colella, María Teresa; Roselló, Arantza; Hartung de Capriles, Claudia; Landaeta, María Eugenia; Olaizola, Carolina; Magaldi, Sylvia

    2005-01-01

    In mycology, culture is the best way to demonstrate and identify pathogenic fungi. Many kinds of media have been developed and modified, using fungal biochemical properties to recognize them. The most common media are Sabouraud, Malt Agar, Cornmeal agar, Potato-dextrose agar, and Staib agar. Staib agar has been widely used for identification of yeasts from the genus Cryptococcus and other fungi. The aim of this study was to demonstrate the usefulness of Staib agar, and of a modified Staib med...

  4. Can smoking cause melanization of Cryptococcus neoformans in vivo?

    Directory of Open Access Journals (Sweden)

    Sabesan, G.

    2011-01-01

    Full Text Available Earlier studies have reported on the ability of Cryptococcus neoformans to synthesize melanin from tobacco extracts / nicotine incorporated in to the medium. However a study on the utilization of components in tobacco smoke by C. neoformans for melanin production was unreported. The present study reports on ability of C. neoformans for melanization using tobacco smoke and therefore substantiate the possible link between smoking and pathogenecity in clinical cryptococcal infections as reported by several researchers.

  5. Meningoradiculitis due to Cryptococcus neofermans in an immunocompetent patient Meningorradiculite lombossacra causada por Cryptococcus neoformans em paciente imunocompetente

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    Leonardo Deus-Silva

    2004-03-01

    Full Text Available Meningoradiculitis refers to combined involvement of meninges and nerve roots. The most frequent location is the lumbosacral region. Etiology is diverse, including inflammatory, infectious and neoplastic disorders. Meningoradiculitis is a rare form of involvement in cryptococcal infection. We describe a case of subacute lower limbs flaccid paresis diagnosed as lumbosacral meningoradiculitis in view of cerebrospinal fluid (CSF inflammatory changes and typical enhancement on MRI of lumbar spine. Cryptococcus neoformans was isolated from CSF. Extensive screening yielded no immunodeficiencies.Meningorradiculite refere-se ao envolvimento simultâneo das meninges e das raízes dos nervos. O local mais freqüentemente acometido é a região lombossacra. Patologias inflamatórias, infecciosas e neoplásicas são as causas mais freqüentes. Meningorradiculite é manifestação rara de infecção por Cryptococcus neoformans. Descrevemos um caso de paresia flácida dos membros inferiores, com diagnóstico de meningorradiculite lombossacra baseado nos achados clínicos, de ressonância magnética da coluna lombar e em alterações inflamatórias do líquido cefalorraqueano (LCR. Avaliação microbiológica do LCR revelou a presença de Cryptococcus neoformans e extensa investigação clínica e laboratorial excluiu imunodeficiências primárias e adquiridas.

  6. Susceptibility testing of Cryptococcus diffluens against amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole and posaconazole

    NARCIS (Netherlands)

    Kantarcioglu, A.S.; Boekhout, T.; Yucel, A.; Altas, K.

    2009-01-01

    Cryptococcus diffluens is a recently re-established species that shares several phenotypic features with Cryptococcus neoformans. We evaluated the application of the Clinical Laboratory Standards Institute (CLSI, formerly NCCLS) macro- and microbroth dilution methods and the E-test agar diffusion

  7. BRONQUIOLITIS RESPIRATORIA Y NEUMONÍA DESCAMATIVA ASOCIADA A FUNGEMIA POR TRICHOSPORON ASAHII EN INMUNOCOMPETENTE: A PROPÓSITO DE UN CASO

    OpenAIRE

    BASTIDAS, ALIRIO R.; PANTOJA, JAIME A.; MUNAR, MARÍA A.; GIRALDO, LUIS F.; YURGAKI, JAMES

    2016-01-01

    Trichosporon asahii es un hongo patógeno emergente reportado en la literatura médica principalmente en pacientes inmunocomprometidos. No obstante, el presente caso es inusual debido a que se trata de un paciente adulto joven inmunocompetente que presentó fungemia por T. asahii y al mismo tiempo desarrolló insuficiencia respiratoria aguda por bronquiolitis respiratoria y neumonía descamativa, la cual resolvió posterior al tratamiento antimicótico instaurado, soporte ventilatorio y vigilancia e...

  8. Mechanisms of Pulmonary Escape and Dissemination by Cryptococcus neoformans

    Directory of Open Access Journals (Sweden)

    Steven T. Denham

    2018-02-01

    Full Text Available Cryptococcus neoformans is a common environmental saprophyte and human fungal pathogen that primarily causes disease in immunocompromised individuals. Similar to many environmentally acquired human fungal pathogens, C. neoformans initiates infection in the lungs. However, the main driver of mortality is invasive cryptococcosis leading to fungal meningitis. After C. neoformans gains a foothold in the lungs, a critical early step in invasion is transversal of the respiratory epithelium. In this review, we summarize current knowledge relating to pulmonary escape. We focus on fungal factors that allow C. neoformans to disseminate from the lungs via intracellular and extracellular routes.

  9. [Fungemia and septic arthritis caused by Saprochaete capitata in a patient with fanconi aplastic anemia: a case report].

    Science.gov (United States)

    Parkan, Ömür Mustafa; Atalay, Mustafa Altay; Koç, Ayşe Nedret; Pala, Çiğdem; Aydemir, Gonca; Kaynar, Leylagül

    2017-01-01

    Saprochaete capitata (formerly known as Blastoschizomyces capitatus, Trichosporon capitatum, Geotrichum capitatum) is a rare but emerging yeast-like fungus. It is commonly found in environmental sources and can be isolated from skin, gastrointestinal system and respiratory tract of healthy individuals as well. It mainly infects patients with hematological malignancies such as acute myeloid leukemia (AML), especially in the presence of neutropenia; and mortality rates are high in those patients. Although the data about the in vitro antifungal susceptibility are limited, it is being reported that amphotericin B and voriconazole are more effective on S.capitata isolates whereas caspofungin had no activity. Here, we report a case of fungemia and septic arthritis due to S.capitata in a patient with Fanconi aplastic anemia. A 22-year-old male patient with Fanconi aplastic anemia was hospitalized in our hematology department for bone marrow transplantation. Two days after the hospitalization, neutropenic fever developed and multiple nodules similar to candidiasis were detected in his liver with the whole abdomen magnetic resonance imaging (MRI). Caspofungin treatment (single 70 mg/kg loading dose, followed by 1 x 50 mg/kg/day) was started. The patient remained febrile, and his blood culture yielded S.capitata. The treatment regimen was changed to a combination of liposomal amphotericin B (3 mg/kg/day) and voriconazole (2 x 4 mg/kg/day). A few days later, pain and swelling came out on patient's left knee and he underwent a surgical process with the prediagnosis of septic arthritis. Culture of synovial fluid was also positive for S.capitata. On the 26th day of the hospitalization, the patient died due to sepsis and multiple organ failure. Patient's blood and synovial fluid samples were incubated in BacT/Alert automated blood culture system (bioMérieux, France). After receiving the growth signal, yeast cells were seen in Gram staining and cream-coloured, wrinkled, yeast

  10. Antifungal and Anti-Biofilm Activity of Essential Oil Active Components against Cryptococcus neoformans and Cryptococcus laurentii

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    Poonam Kumari

    2017-11-01

    Full Text Available Cryptococcosis is an emerging and recalcitrant systemic infection occurring in immunocompromised patients. This invasive fungal infection is difficult to treat due to the ability of Cryptococcus neoformans and Cryptococcus laurentii to form biofilms resistant to standard antifungal treatment. The toxicity concern of these drugs has stimulated the search for natural therapeutic alternatives. Essential oil and their active components (EO-ACs have shown to possess the variety of biological and pharmacological properties. In the present investigation the effect of six (EO-ACs sourced from Oregano oil (Carvacrol, Cinnamon oil (Cinnamaldehyde, Lemongrass oil (Citral, Clove oil (Eugenol, Peppermint oil (Menthol and Thyme oil (thymol against three infectious forms; planktonic cells, biofilm formation and preformed biofilm of C. neoformans and C. laurentii were evaluated as compared to standard drugs. Data showed that antibiofilm activity of the tested EO-ACs were in the order: thymol>carvacrol>citral>eugenol=cinnamaldehyde>menthol respectively. The three most potent EO-ACs, thymol, carvacrol, and citral showed excellent antibiofilm activity at a much lower concentration against C. laurentii in comparison to C. neoformans indicating the resistant nature of the latter. Effect of the potent EO-ACs on the biofilm morphology was visualized using scanning electron microscopy (SEM and confocal laser scanning microscopy (CLSM, which revealed the absence of extracellular polymeric matrix (EPM, reduction in cellular density and alteration in the surface morphology of biofilm cells. Further, to realize the efficacy of the EO-ACs in terms of human safety, cytotoxicity assays and co-culture model were evaluated. Thymol and carvacrol as compared to citral were the most efficient in terms of human safety in keratinocyte- Cryptococcus sp. co-culture infection model suggesting that these two can be further exploited as cost-effective and non-toxic anti

  11. Outbreak of Fusarium oxysporum infections in children with cancer: an experience with 7 episodes of catheter-related fungemia.

    Science.gov (United States)

    Carlesse, Fabianne; Amaral, Anna-Paula C; Gonçalves, Sarah S; Xafranski, Hemilio; Lee, Maria-Lucia M; Zecchin, Victor; Petrilli, Antonio S; Al-Hatmi, Abdullah M; Hagen, Ferry; Meis, Jacques F; Colombo, Arnaldo L

    2017-01-01

    Fusarium species are widely spread in nature as plant pathogens but are also able to cause opportunistic fungal infections in humans. We report a cluster of Fusarium oxysporum bloodstream infections in a single pediatric cancer center. All clinical and epidemiological data related to an outbreak involving seven cases of fungemia by Fusarium oxysporum during October 2013 and February 2014 were analysed. All cultured isolates ( n  = 14) were identified to species level by sequencing of the TEF1 and RPB2 genes. Genotyping of the outbreak isolates was performed by amplified fragment length polymorphism fingerprinting. In a 5-month period 7 febrile pediatric cancer patients were diagnosed with catheter-related Fusarium oxysporum bloodstream infections. In a time span of 11 years, only 6 other infections due to Fusarium were documented and all were caused by a different species, Fusarium solani . None of the pediatric cancer patients had neutropenia at the time of diagnosis and all became febrile within two days after catheter manipulation in a specially designed room. Extensive environmental sampling in this room and the hospital did not gave a clue to the source. The outbreak was terminated after implementation of a multidisciplinary central line insertion care bundle. All Fusarium strains from blood and catheter tips were genetically related by amplified fragment length polymorphism fingerprinting. All patients survived the infection after prompt catheter removal and antifungal therapy. A cluster with, genotypical identical, Fusarium oxysporum strains infecting 7 children with cancer, was most probably catheter-related. The environmental source was not discovered but strict infection control measures and catheter care terminated the outbreak.

  12. Outbreak of Fusarium oxysporum infections in children with cancer: an experience with 7 episodes of catheter-related fungemia

    Directory of Open Access Journals (Sweden)

    Fabianne Carlesse

    2017-09-01

    Full Text Available Abstract Background Fusarium species are widely spread in nature as plant pathogens but are also able to cause opportunistic fungal infections in humans. We report a cluster of Fusarium oxysporum bloodstream infections in a single pediatric cancer center. Methods All clinical and epidemiological data related to an outbreak involving seven cases of fungemia by Fusarium oxysporum during October 2013 and February 2014 were analysed. All cultured isolates (n = 14 were identified to species level by sequencing of the TEF1 and RPB2 genes. Genotyping of the outbreak isolates was performed by amplified fragment length polymorphism fingerprinting. Results In a 5-month period 7 febrile pediatric cancer patients were diagnosed with catheter-related Fusarium oxysporum bloodstream infections. In a time span of 11 years, only 6 other infections due to Fusarium were documented and all were caused by a different species, Fusarium solani. None of the pediatric cancer patients had neutropenia at the time of diagnosis and all became febrile within two days after catheter manipulation in a specially designed room. Extensive environmental sampling in this room and the hospital did not gave a clue to the source. The outbreak was terminated after implementation of a multidisciplinary central line insertion care bundle. All Fusarium strains from blood and catheter tips were genetically related by amplified fragment length polymorphism fingerprinting. All patients survived the infection after prompt catheter removal and antifungal therapy. Conclusion A cluster with, genotypical identical, Fusarium oxysporum strains infecting 7 children with cancer, was most probably catheter-related. The environmental source was not discovered but strict infection control measures and catheter care terminated the outbreak.

  13. Cryptococcus neoformans infective endocarditis of native valves in an immunocompetent host

    Directory of Open Access Journals (Sweden)

    Moni Roy

    Full Text Available With the emergence of Human immunodeficiency virus (HIV and the resulting immunocompromised state, Cryptococcus neoformans infections have gained more importance in clinical practice. Cryptococcal infections in immunocompetent hosts continue to be uncommon. We present a rare case of Cryptococcus neoformans infective endocarditis (IE in a young immunocompetent male. As per our literature review, this is the first reported case of native valve Cryptococcus neoformans endocarditis in an immunocompetent host. All cases till date have been reported in patients with underlying immunocompromised state or prosthetic valve.

  14. Neurocriptococose por Cryptococcus neoformans não capsulado

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    C.S. Lacaz

    1993-09-01

    Full Text Available Os autores registram um caso de neurocriptococose em paciente HIV-negativo, por Cryptococcus neoformans acapsulado ou deficiente em cápsula. O quadro neurológico era de meningoencefalite subaguda, compatível ao diagnóstico de neurotuberculose, pelo exame do líquido cefalorraqueano (LCR, Estruturas leveduriformes foram encontradas no interior de macrófagos, ao exame citomorfológico do LCR. Cultivo do sedimento do LCR revelou a presença de Cryptococcus neoformans não capsulado (identificação bioquímica. A inoculação da amostra em camundongo, por via intraperitoneal, permitiu a produção de cápsula que desaparecia em cultivos. Foi estudada a micromorfologia do fungo à microscopia eletrônica de varredura. A evolução foi favorável com o emprego da anfotericina B associada a 5-fluorocitosina. Não foi caracterizada a variadade de Criptococcus neoformans agente do processo.

  15. Cryptococcus neoformans var neoformans isolated from droppings of captive birds in Nigeria.

    Science.gov (United States)

    Irokanulo, E O; Makinde, A A; Akuesgi, C O; Ekwonu, M

    1997-04-01

    The yeast, Cryptococcus neoformans, was found in apparently healthy birds at the Jos Wildlife Park and Zoo in Jos, Nigeria. Cryptococcus neoformans var neoformans was isolated from feces of four captive bird species. Five isolates belonged to serotype A while two were serotype D. Serotype A of C. neoformans was isolated from a white face duck (Dendrocygna viduata), eagle owl (Bubo africanus cinerascene) and peacock (Pavo cristatus). The other two (serotype D), were isolated from a spotted eagle owl.

  16. Two rapid pigmentation tests for identification of Cryptococcus neoformans.

    Science.gov (United States)

    Kaufmann, C S; Merz, W G

    1982-01-01

    Two tests were developed for the rapid identification of Cryptococcus neoformans based on pigment produced by the organism's phenoloxidase activity. Caffeic acid was incorporated into cornmeal agar, a medium used routinely for yeast identification. When tested on this medium, only C. neoformans isolates produced brown pigment. All other yeasts maintained their normal morphology and did not produce the reaction product. A non-medium-based test was developed for same-day identification of C. neoformans isolates. Paper strips saturated with a buffered L-beta-3,4-dihydroxyphenylalanine-ferric citrate solution were inoculated with isolates and incubated at 37 degrees C. Pigment production occurred only with C. neoformans isolates, many within 60 to 90 min. All other yeasts remained negative. PMID:7040452

  17. The first reported case of canine subcutaneous Cryptococcus flavescens infection.

    Science.gov (United States)

    Kano, Rui; Ishida, Rinei; Nakane, Shinsuke; Sekiguchi, Maiko; Hasegawa, Atsuhiko; Kamata, Hiroshi

    2012-03-01

    This report describes the first documented case of subcutaneous infection due to Cryptococcus flavescens in a dog. The chief symptoms of the patient dog were abscessed lesions on the dorsal muzzle, right eyelid, and lower jaw. Biopsy specimens from the lesions on the dorsal muzzle and lower jaw showed pyogranulomatous inflammation with numerous yeast cells. The patient dog was diagnosed with a subcutaneous fungal infection and orally received 5 mg/kg itraconazole once a day for 2 months, the abscesses disappeared. After 1 month at the end of treatment, the skin lesions did not redevelop. Isolates from the biopsy specimens were identified as C. flavescens by molecular analysis as well as morphologic and biochemical examination, indicating that C. flavescens is a potential canine pathogen.

  18. Biodiesel production from yeast Cryptococcus sp. using Jerusalem artichoke.

    Science.gov (United States)

    Sung, Mina; Seo, Yeong Hwan; Han, Shin; Han, Jong-In

    2014-03-01

    Jerusalem artichoke was investigated as a cheap substrate for the heterotrophic production using a lab yeast strain Cryptococcus sp. Using Response Surface Method, 54.0% of fructose yield was achieved at 12% of dried Jerusalem artichoke powder, 0.57% of nitric acid concentration, 117°C of reaction temperature, and 49min of reaction time. At this optimal condition, nitric acid showed the best catalytic activity toward inulin hydrolysis and also the resulting fructose hydrolyte supported the highest microbial growth compared with other acids. In addition, lipid productivity of 1.73g/L/d was achieved, which is higher than a defined medium using pure fructose as a substrate. Lipid quality was also found to be generally satisfactory as a feedstock for fuel, demonstrating Jerusalem artichoke could indeed be a good and cheap option for the purpose of biodiesel production. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Fenoloxidasa Modificada: Clave para identificar cepas de Cryptococcus neoformans

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    Carlos Canelo D

    2000-01-01

    Full Text Available Cryptococcus neoformans es la única levadura patógena capaz de sintetizar pigmentos como la melanina mediante la actividad de su enzima llamada fenoloxidasa. El objetivo del presente estudio fue implementar y estandarizar la prueba de la fenoloxidasa, como técnica complementaria en la identificación de cepas de C. neoformans. Se estudiaron 21 cepas, identificadas previamente con métodos convencionales. La prueba de la fenoloxidasa fue modificada debido a que su empleo originaba 9,6% (2/21 de falsa negatividad. Esta prueba modificada se optimizó a 28°C a partir de un medio con baja concentración de glucosa. Ningún aislamiento falso negativo fue encontrado luego de repetir tres veces el ensayo, y el pigmento melanina fue detectado con mayor rapidez.

  20. [A comparative study of blood culture conventional method vs. a modified lysis/centrifugation technique for the diagnosis of fungemias].

    Science.gov (United States)

    Santiago, Axel Rodolfo; Hernández, Betsy; Rodríguez, Marina; Romero, Hilda

    2004-12-01

    The purpose of this work was to compare the efficacy of blood culture conventional method vs. a modified lysis/centrifugation technique. Out of 450 blood specimens received in one year, 100 where chosen for this comparative study: 60 from patients with AIDS, 15 from leukemic patients, ten from febrile neutropenic patients, five from patients with respiratory infections, five from diabetics and five from septicemic patients. The specimens were processed, simultaneously, according to the above mentioned methodologies with daily inspections searching for fungal growth in order to obtain the final identification of the causative agent. The number (40) of isolates recovered was the same using both methods, which included; 18 Candida albicans (45%), ten Candida spp. (25%), ten Histoplasma capsulatum (25%), and two Cryptococcus neoformans (5%). When the fungal growth time was compared by both methods, growth was more rapid when using the modified lysis/centrifugation technique than when using the conventional method. Statistical analysis revealed a significant difference (pcentrifugation technique showed to be more efficacious than the conventional one, and therefore the implementation of this methodology is highly recommended for the isolation of fungi from blood.

  1. Cryptococcus haglerorum, sp. nov., an anamorphic basidiomycetous yeast isolated from nests of the leaf-cutting ant Atta sexdens.

    NARCIS (Netherlands)

    Middelhoven, W.J.; Fonseca, A.; Carreiro, S.C.; Pagnocca, F.C.; Bueno, O.C.

    2003-01-01

    A yeast strain (CBS 8902) was isolated from the nest of a leaf-cutting ant and was shown to be related to Cryptococcus humicola. Sequencing of the D1/D2 region of the 26S ribosomal DNA and physiological characterization revealed a separate taxonomic position. A novel species named Cryptococcus

  2. CARACTERIZACION FENOTIPICA Y TIPIFICACION MOLECULAR DE CEPAS DE CRYPTOCOCCUS NEOFORMANS AISLADAS DE MUESTRAS CLINICAS EN HOSPITALES TERCIARIOS DEL SUR DE CHILE

    OpenAIRE

    BARRIA I., YOLANDA B.

    2012-01-01

    Criptococosis es una micosis sistémica oportunista causada por dos especies levaduriformes capsuladas, C. neoformans causan infecciones en individuos immunodeprimidos ptincipalmente y Criptococcus gattii lo hace tanto en immunocompetentes como en personas inmunodeprimidas. C. neoformans es comúnmente asociado a excretas de palomas mientras que la especie C. gattii es encontrado en especies arbóreas como eucaliptos, ficus y otras. C neoformcms se subdivide en dos variedades distintas neofor...

  3. The link between morphotype transition and virulence in Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Linqi Wang

    Full Text Available Cryptococcus neoformans is a ubiquitous human fungal pathogen. This pathogen can undergo morphotype transition between the yeast and the filamentous form and such morphological transition has been implicated in virulence for decades. Morphotype transition is typically observed during mating, which is governed by pheromone signaling. Paradoxically, components specific to the pheromone signaling pathways play no or minimal direct roles in virulence. Thus, the link between morphotype transition and virulence and the underlying molecular mechanism remain elusive. Here, we demonstrate that filamentation can occur independent of pheromone signaling and mating, and both mating-dependent and mating-independent morphotype transition require the transcription factor Znf2. High expression of Znf2 is necessary and sufficient to initiate and maintain sex-independent filamentous growth under host-relevant conditions in vitro and during infection. Importantly, ZNF2 overexpression abolishes fungal virulence in murine models of cryptococcosis. Thus, Znf2 bridges the sex-independent morphotype transition and fungal pathogenicity. The impacts of Znf2 on morphological switch and pathogenicity are at least partly mediated through its effects on cell adhesion property. Cfl1, a Znf2 downstream factor, regulates morphogenesis, cell adhesion, biofilm formation, and virulence. Cfl1 is the first adhesin discovered in the phylum Basidiomycota of the Kingdom Fungi. Together with previous findings in other eukaryotic pathogens, our findings support a convergent evolution of plasticity in morphology and its impact on cell adhesion as a critical adaptive trait for pathogenesis.

  4. Hydroxyurea treatment inhibits proliferation of Cryptococcus neoformans in mice

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    Kaushlendra eTripathi

    2012-05-01

    Full Text Available The fungal pathogen Cryptococcus neoformans (Cn is a serious threat to immunocompromised individuals, especially for HIV patients who develop meningoencephalitis. For effective cryptococcal treatment, novel antifungal drugs or innovative combination therapies are needed. Recently, sphingolipids have emerged as important bioactive molecules in the regulation of microbial pathogenesis. Previously we reported that the sphingolipid pathway gene, ISC1, which is responsible for ceramide production, is a major virulence factor in Cn infection. Here we report our studies of the role of ISC1 during genotoxic stress induced by the antineoplastic hydroxyurea (HU and methylmethane sulfonate (MMS, which affect DNA replication and genome integrity. We observed that Cn cells lacking ISC1 are highly sensitive to HU and MMS in a rich culture medium. HU affected cell division of Cn cells lacking the ISC1 gene, resulting in cell clusters. Cn ISC1, when expressed in a Saccharomyces cerevisiae (Sc strain lacking its own ISC1 gene, restored HU resistance. In macrophage-like cells, although HU affected the proliferation of WT Cn cells by 50% at the concentration tested, HU completely inhibited Cn isc1-delta cell proliferation. Interestingly, our preliminary data show that mice infected with WT or Cn isc1-delta cells and subsequently treated with HU had longer lifespans than untreated, infected control mice. Our work suggests that the sphingolipid pathway gene, ISC1, is a likely target for combination therapy with traditional drugs such as HU.

  5. Terbinafine inhibits Cryptococcus neoformans growth and modulates fungal morphology

    Directory of Open Access Journals (Sweden)

    Caroline Rezende Guerra

    2012-08-01

    Full Text Available Cryptococcus neoformans is an encapsulated fungus that causes cryptococcosis. Central nervous system infection is the most common clinical presentation followed by pulmonary, skin and eye manifestations. Cryptococcosis is primarily treated with amphotericin B (AMB, fluconazole (FLC and itraconazole (ITC. In the present work, we evaluated the in vitro effect of terbinafine (TRB, an antifungal not commonly used to treat cryptococcosis. We specifically examined the effects of TRB, either alone or in conjunction with AMB, FLC and ITC, on clinical C. neoformans isolates, including some isolates resistant to AMB and ITC. Broth microdilution assays showed that TRB was the most effective drug in vitro. Antifungal combinations demonstrated synergism of TRB with AMB, FLC and ITC. The drug concentrations used for the combination formulations were as much as 32 and 16-fold lower than the minimum inhibitory concentration (MIC values of FLC and AMB alone, respectively. In addition, calcofluor white staining revealed the presence of true septa in hyphae structures that were generated after drug treatment. Ultrastructural analyses demonstrated several alterations in response to drug treatment, such as cell wall alterations, plasma membrane detachment, presence of several cytoplasmic vacuoles and mitochondrial swelling. Therefore, we believe that the use of TRB alone or in combination with AMB and azoles should be explored as an alternative treatment for cryptococcosis patients who do not respond to standard therapies.

  6. Extracellular glycosylphosphatidylinositol-anchored mannoproteins and proteases of Cryptococcus neoformans.

    Science.gov (United States)

    Eigenheer, Richard A; Jin Lee, Young; Blumwald, Eduardo; Phinney, Brett S; Gelli, Angie

    2007-06-01

    Extracellular proteins of Cryptococcus neoformans are involved in the pathogenesis of cryptococcosis, and some are immunoreactive antigens that may potentially serve as candidates for vaccine development. To further study the extracellular proteome of the human fungal pathogen Cry. neoformans, we conducted a proteomic analysis of secreted and cell wall-bound proteins with an acapsular strain of Cry. neoformans. Proteins were identified from both intact cells and cell walls. In both cases, extracellular proteins were removed with trypsin or beta-glucanase, and then all proteins/peptides were purified by solid-phase extraction, spin dialysis, and HPLC, and identified by liquid chromatography-mass spectrometry. This study identified 29 extracellular proteins with a predicted N-terminal signal sequence and also a predicted glycosylphosphatidylinositol anchor motif in more than half. Among the novel proteins identified were five glycosylphosphatidylinositol-anchored proteins with extensive Ser/Thr-rich regions but no apparent functional domains, a glycosylphosphatidylinositol-anchored aspartic protease, and a metalloprotease with structural similarity to an elastinolytic metalloprotease of Aspergillus fumigatus. This study suggests that Cry. neoformans has the machinery required to target glycosylphosphatidylinositol-anchored proteins to the cell wall, and it confirms the extracellular proteolytic ability of Cry. neoformans.

  7. Cryptococcus neoformans: Tripping on Acid in the phagolysosome

    Directory of Open Access Journals (Sweden)

    Carlos Miguel De Leon Rodriguez

    2016-02-01

    Full Text Available Cryptococcus neoformans (Cn is a basidiomycetous pathogenic yeast that is a frequent cause of meningoencephalitis in immunocompromised individuals. Cn is a facultative intracellular pathogen in mammals, insects and amoeba. Cn infection occurs after inhalation of spores or desiccated cells from the environment. After inhalation Cn localizes to the lungs where it can be phagocytosed by alveolar macrophages. Cn is surrounded by a polysaccharide capsule that helps the fungus survive in vivo by interfering with phagocytosis, quenching free radical bursts and shedding polysaccharides that negatively modulates the immune system. After phagocytosis, Cn resides within the phagosome that matures to become a phagolysosome, a process that results in the acidification of the phagolysosomal lumen. Cn replicates at a higher rate inside macrophages than in the extracellular environment, possibly as a result that the phagosomal pH is near that optimal for growth. Cn increases the phagolysosomal pH and modulates the dynamics of Rab GTPases interaction with the phagolysosome. Chemical manipulation of the phagolysosomal pH with drugs can result in direct and indirect killing of Cn and reduced non-lytic exocytosis. Phagolysosomal membrane damage after Cn infection occurs both in vivo and in vitro, and is required for Cn growth and survival. Macrophage treatment with IFN-γ reduces the phagolysosomal damage and increases intracellular killing of Cn. Studies on mice and humans show that treatment with IFN-γ can improve host control of the disease. However the mechanism by which Cn mediates phagolysosomal membrane damage remains unknown but likely candidates are phospholipases and mechanical damage from an enlarging capsule. Here we review Cn intracellular interaction with a particular emphasis on phagosomal interactions and develop the notion that the extent of damage of the phagosomal membrane is a key determinant of the outcome of the Cn-macrophage interaction.

  8. Toward an integrated model of capsule regulation in Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Brian C Haynes

    2011-12-01

    Full Text Available Cryptococcus neoformans is an opportunistic fungal pathogen that causes serious human disease in immunocompromised populations. Its polysaccharide capsule is a key virulence factor which is regulated in response to growth conditions, becoming enlarged in the context of infection. We used microarray analysis of cells stimulated to form capsule over a range of growth conditions to identify a transcriptional signature associated with capsule enlargement. The signature contains 880 genes, is enriched for genes encoding known capsule regulators, and includes many uncharacterized sequences. One uncharacterized sequence encodes a novel regulator of capsule and of fungal virulence. This factor is a homolog of the yeast protein Ada2, a member of the Spt-Ada-Gcn5 Acetyltransferase (SAGA complex that regulates transcription of stress response genes via histone acetylation. Consistent with this homology, the C. neoformans null mutant exhibits reduced histone H3 lysine 9 acetylation. It is also defective in response to a variety of stress conditions, demonstrating phenotypes that overlap with, but are not identical to, those of other fungi with altered SAGA complexes. The mutant also exhibits significant defects in sexual development and virulence. To establish the role of Ada2 in the broader network of capsule regulation we performed RNA-Seq on strains lacking either Ada2 or one of two other capsule regulators: Cir1 and Nrg1. Analysis of the results suggested that Ada2 functions downstream of both Cir1 and Nrg1 via components of the high osmolarity glycerol (HOG pathway. To identify direct targets of Ada2, we performed ChIP-Seq analysis of histone acetylation in the Ada2 null mutant. These studies supported the role of Ada2 in the direct regulation of capsule and mating responses and suggested that it may also play a direct role in regulating capsule-independent antiphagocytic virulence factors. These results validate our experimental approach to dissecting

  9. PERIORBITAL NECROTIZING FASCIITIS DUE TO CRYPTOCOCCUS NEOFORMANS IN A HEALTHY-YOUNG MAN

    NARCIS (Netherlands)

    DOORENBOSBOT, ACC; HOOYMANS, JMM; BLANKSMA, LJ

    1990-01-01

    A case report is presented of a healthy 25-year-old man who developed a periorbital necrotising fasciitis after a trivial trauma with a wooden splinter. Necrotising fasciitis of the eyelids occurs rarely. Cryptococcus neoformans is not described as a causative factor of necrotizing fasciitis.

  10. Wood and bark anatomy of young beech in relation to Cryptococcus attack

    Science.gov (United States)

    David. Lonsdale

    1983-01-01

    Within a sample of European beech, partial resistance to attack by the beech scale, Cryptococcus fagisuga, was associated with a smooth bark which had a regular, vertical pattern in its surface 'growth lines'. Such bark contained relatively little lignified outer parenchyma, and the main stone cell layer was strongly developed. The '...

  11. Cryptococcus sp abdominal infection in a patient with acquired immunodeficiency under secondary prophylaxis with fluconazole; Infeccao abdominal por Cryptococcus sp em paciente com imunodeficiencia adquirida em uso de profilaxia secundaria com fluconazol

    Energy Technology Data Exchange (ETDEWEB)

    Coutinho, Rosane Luiza; Zarehdinne, Monica [Infectologia do Hospital Eduardo de Menezes Belo Horizonte, MG (Brazil); Peixoto, Arley; Tardieu Junior, Jose [Medicina da Faculdade de Saude e Ecologia Humana - FASEH, Vespasiano, MG (Brazil); Pedroso, Enio Roberto Pietra [Departamento de Clinica Medica da Faculdade de Medicina da Universidade Federal de Minas Gerais - UFMG, Belo Horizonte, MG (Brazil)

    2011-10-15

    This paper reports on a Cryptococcus sp infection relapse in a patient with acquired immunodeficiency three years after the first episode, when her T CD4+ lymphocyte count was high and she was making regular use of antiretroviral drugs. (author)

  12. Modulation of Replicative Lifespan in Cryptococcus neoformans: Implications for Virulence

    Science.gov (United States)

    Bouklas, Tejas; Jain, Neena; Fries, Bettina C.

    2017-01-01

    The fungal pathogen, Cryptococcus neoformans, has been shown to undergo replicative aging. Old cells are characterized by advanced generational age and phenotypic changes that appear to mediate enhanced resistance to host and antifungal-based killing. As a consequence of this age-associated resilience, old cells accumulate during chronic infection. Based on these findings, we hypothesized that shifting the generational age of a pathogenic yeast population would alter its vulnerability to the host and affect its virulence. SIR2 is a well-conserved histone deacetylase, and a pivotal target for the development of anti-aging drugs. We tested its effect on C. neoformans’ replicative lifespan (RLS). First, a mutant C. neoformans strain (sir2Δ) was generated, and confirmed a predicted shortened RLS in sir2Δ cells consistent with its known role in aging. Next, RLS analysis showed that treatment of C. neoformans with Sir2p-agonists resulted in a significantly prolonged RLS, whereas treatment with a Sir2p-antagonist shortened RLS. RLS modulating effects were dependent on SIR2 and not observed in sir2Δ cells. Because SIR2 loss resulted in a slightly impaired fitness, effects of genetic RLS modulation on virulence could not be compared with wild type cells. Instead we chose to chemically modulate RLS, and investigated the effect of Sir2p modulating drugs on C. neoformans cells in a Galleria mellonella infection model. Consistent with our hypothesis that shifts in the generational age of the infecting yeast population alters its vulnerability to host cells, we observed decreased virulence of C. neoformans in the Galleria host when RLS was prolonged by treatment with Sir2p agonists. In contrast, treatment with a Sir2p antagonist, which shortens RLS enhanced virulence in Galleria. In addition, combination of Sir2p agonists with antifungal therapy enhanced the antifungal’s effect. Importantly, no difference in virulence was observed with drug treatment when sir2Δ cells

  13. Susceptibilidad "in vitro" de cepas de Cryptococcus a 5 drogas antifungicas "In vitro" susceptibility of Cryptococcus strains to 5 antifungal drugs

    Directory of Open Access Journals (Sweden)

    A. J. Bava

    1989-10-01

    Full Text Available Se estudió la susceptibilidad "in vitro" de 24 cepas de 3 especies del género Cryptococcus a 5 drogas antifúngicas (anfotericina B, 5 fluorocitosina, ketoconazol, itraconazol y miconazol. Las mismas se agruparon según su especie, variedad y origen de aislamiento. Para determinar la concentración inhibitoria mínima (C.I.M. de cada droga se empleó el método de dilución en agar con el medio básico nitrogenado para levaduras, adicionado de glucosa. Se obtuvo además la media geométrica de estos valores para cada grupo y se comparó cada uno de ellos. Los resultados obtenidos fueron homogéneos con la sola excepción de las cepas de Cryptococcus sp (no neoformans, en las cuales se detectaron elevados valores de C.I.M. para la 5 fluorocitosina.A comparative study of the "in vitro" susceptibility of 24 Cryptococcus strains to 5 antifungal drugs (amphotericin B, 5 fluorocytosine, miconazole, itraconazole and ketoconazole, was carried out. These strains were grouped according to species, varieties and isolation's origins. The minimum inhibitory concentration (M.I.C. was determinated by the agar dilution technique in yeast nitrogen base agar with dextrose. The mean geometrical of the M.I.C. values of each group was compared with the others. The results obtained were homogeneous with the only exception of the "non neoformans" strains, in which, higher M.I.C. to 5 fluorocytosine values were detected.

  14. Life History Responses and Gene Expression Profiles of the Nematode Pristionchus pacificus Cultured on Cryptococcus Yeasts.

    Directory of Open Access Journals (Sweden)

    Gaurav V Sanghvi

    Full Text Available Nematodes, the earth's most abundant metazoa are found in all ecosystems. In order to survive in diverse environments, they have evolved distinct feeding strategies and they can use different food sources. While some nematodes are specialists, including parasites of plants and animals, others such as Pristionchus pacificus are omnivorous feeders, which can live on a diet of bacteria, protozoans, fungi or yeast. In the wild, P. pacificus is often found in a necromenic association with beetles and is known to be able to feed on a variety of microbes as well as on nematode prey. However, in laboratory studies Escherichia coli OP50 has been used as standard food source, similar to investigations in Caenorhabditis elegans and it is unclear to what extent this biases the obtained results and how relevant findings are in real nature. To gain first insight into the variation in traits induced by a non-bacterial food source, we study Pristionchus-fungi interactions under laboratory conditions. After screening different yeast strains, we were able to maintain P. pacificus for at least 50-60 generations on Cryptococcus albidus and Cryptococcus curvatus. We describe life history traits of P. pacificus on both yeast strains, including developmental timing, survival and brood size. Despite a slight developmental delay and problems to digest yeast cells, which are both reflected at a transcriptomic level, all analyses support the potential of Cryptococcus strains as food source for P. pacificus. In summary, our work establishes two Cryptococcus strains as alternative food source for P. pacificus and shows change in various developmental, physiological and morphological traits, including the transcriptomic profiles.

  15. Detection of Cryptococcus neoformans DNA in Tissue Samples by Nested and Real-Time PCR Assays

    OpenAIRE

    Bialek, Ralf; Weiss, Michael; Bekure-Nemariam, Kubrom; Najvar, Laura K.; Alberdi, Maria B.; Graybill, John R.; Reischl, Udo

    2002-01-01

    Two PCR protocols targeting the 18S rRNA gene of Cryptococcus neoformans were established, compared, and evaluated in murine cryptococcal meningitis. One protocol was designed as a nested PCR to be performed in conventional block thermal cyclers. The other protocol was designed as a quantitative single-round PCR adapted to LightCycler technology. One hundred brain homogenates and dilutions originating from 20 ICR mice treated with different azoles were examined. A fungal burden of 3 × 101 to ...

  16. Synergy and antagonism between iron chelators and antifungal drugs in Cryptococcus.

    Science.gov (United States)

    Lai, Yu-Wen; Campbell, Leona T; Wilkins, Marc R; Pang, Chi Nam Ignatius; Chen, Sharon; Carter, Dee A

    2016-10-01

    Fungal infections remain very difficult to treat, and developing new antifungal drugs is difficult and expensive. Recent approaches therefore seek to augment existing antifungals with synergistic agents that can lower the therapeutic dose, increase efficacy and prevent resistance from developing. Iron limitation can inhibit microbial growth, and iron chelators have been employed to treat fungal infections. In this study, chequerboard testing was used to explore combinations of iron chelators with antifungal agents against pathogenic Cryptococcus spp. with the aim of determining how disruption to iron homeostasis affects antifungal susceptibility. The iron chelators ethylenediaminetetraacetic acid (EDTA), deferoxamine (DFO), deferiprone (DFP), deferasirox (DSX), ciclopirox olamine and lactoferrin (LF) were paired with the antifungal agents amphotericin B (AmB), fluconazole, itraconazole, voriconazole and caspofungin. All chelators except for DFO increased the efficacy of AmB, and significant synergy was seen between AmB and LF for all Cryptococcus strains. Addition of exogenous iron rescued cells from the antifungal effect of LF alone but could not prevent inhibition by AmB + LF, indicating that synergy was not due primarily to iron chelation but to other properties of LF that were potentiated in the presence of AmB. Significant synergy was not seen consistently for other antifungal-chelator combinations, and EDTA, DSX and DFP antagonised the activity of azole drugs in strains of Cryptococcus neoformans var. grubii. This study highlights the range of interactions that can be induced by chelators and indicates that most antifungal drugs are not enhanced by iron limitation in Cryptococcus. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  17. Microstructure of Cell Wall-Associated Melanin in the Human Pathogenic Fungus cryptococcus neoformans

    OpenAIRE

    Eisenman, H.C.; Nosanchuk, J.D.; Webber, J. Beau W.; Emerson, T.A.; Casadevall, A.

    2005-01-01

    Melanin is a virulence factor for many pathogenic fungal species,including Cryptococcus neoformans. Melanin is deposited in the cell wall, and melanin isolated from this fungus retains the shape of the cells, resulting in hollow spheres called ``ghosts''. In this study, atomic force, scanning electron, and transmission electron microscopy revealed that melanin ghosts are covered with roughly spherical granular particles approximately 40-130 nm in diameter, and that the melanin is arranged in ...

  18. The Genome of the Basidiomycetous Yeast and Human Pathogen Cryptococcus neoformans

    OpenAIRE

    Loftus, Brendan J.; Fung, Eula; Roncaglia, Paola; Rowley, Don; Amedeo, Paolo; Bruno, Dan; Vamathevan, Jessica; Miranda, Molly; Anderson, Iain J.; Fraser, James A.; Allen, Jonathan E.; Bosdet, Ian E.; Brent, Michael R.; Chiu, Readman; Doering, Tamara L.

    2005-01-01

    Cryptococcus neoformans is a basidiomycetous yeast ubiquitous in the environment, a model for fungal pathogenesis, and an opportunistic human pathogen of global importance. We have sequenced its ~20-megabase genome, which contains ~6500 intron-rich gene structures and encodes a transcriptome abundant in alternatively spliced and antisense messages. The genome is rich in transposons, many of which cluster at candidate centromeric regions. The presence of these transposons may drive karyotype i...

  19. Pleiotropic effects of deubiquitinating enzyme Ubp5 on growth and pathogenesis of Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Wei Fang

    Full Text Available Ubiquitination is a reversible protein modification that influences various cellular processes in eukaryotic cells. Deubiquitinating enzymes remove ubiquitin, maintain ubiquitin homeostasis and regulate protein degradation via the ubiquitination pathway. Cryptococcus neoformans is an important basidiomycete pathogen that causes life-threatening meningoencephalitis primarily in the immunocompromised population. In order to understand the possible influence deubiquitinases have on growth and virulence of the model pathogenic yeast Cryptococcus neoformans, we generated deletion mutants of seven putative deubiquitinase genes. Compared to other deubiquitinating enzyme mutants, a ubp5Δ mutant exhibited severely attenuated virulence and many distinct phenotypes, including decreased capsule formation, hypomelanization, defective sporulation, and elevated sensitivity to several external stressors (such as high temperature, oxidative and nitrosative stresses, high salts, and antifungal agents. Ubp5 is likely the major deubiquitinating enzyme for stress responses in C. neoformans, which further delineates the evolutionary divergence of Cryptococcus from the model yeast S. cerevisiae, and provides an important paradigm for understanding the potential role of deubiquitination in virulence by other pathogenic fungi. Other putative deubiquitinase mutants (doa4Δ and ubp13Δ share some phenotypes with the ubp5Δ mutant, illustrating functional overlap among deubiquitinating enzymes in C. neoformans. Therefore, deubiquitinating enzymes (especially Ubp5 are essential for the virulence composite of C. neoformans and provide an additional yeast survival and propagation advantage in the host.

  20. Evaluation of SOC for the presumptive identification of Candida albicans and Cryptococcus neoformans.

    Science.gov (United States)

    Fleming, W H; Knezek, K L; Dorn, G L

    1987-01-01

    SOC, a fungal growth medium composed of Solryth, oxgall, and caffeic acid, was evaluated as a medium to provide rapid, differential identification of Candida albicans and Cryptococcus neoformans. Using a variety of common isolation media to produce the yeast inocula, the germ tube methods tested ranked in the following order of decreasing sensitivity: SOC (97% +/- 1), serum (92% +/- 5), rabbit coagulase plasma with EDTA in combination with tryptic soy broth (89% +/- 5), TOC (89% +/- 6), and rabbit coagulase plasma with EDTA (83% +/- 4). In chlamydospore production, SOC also proved to be the most sensitive after 24 h incubation: SOC (96% +/- 2), TOC (80% +/- 2), and cornmeal-Tween 80 agar (14% +/- 3). Other medically important yeasts showed normal patterns of growth within 24 h on SOC, thus assisting in their identification. Eighty strains of Cryptococcus neoformans showed characteristic brown pigmentation on SOC and TOC within 18 h, while all other species of the genus Cryptococcus and 229 Candida isolates did not show a change in pigmentation.

  1. Molecular identity and prevalence of Cryptococcus spp. nasal carriage in asymptomatic feral cats in Italy.

    Science.gov (United States)

    Danesi, Patrizia; Furnari, Carmelo; Granato, Anna; Schivo, Alice; Otranto, Domenico; Capelli, Gioia; Cafarchia, Claudia

    2014-10-01

    Cryptococcosis is a life-threatening fungal disease that infects humans and animals worldwide. Inhalation of fungal particles from an environmental source can cause primary infection of the respiratory system. As animals can be considered a sentinel for human diseases, the aim of this study was to determine the prevalence and molecular identity of Cryptococcus spp. in the nasal cavity of feral cats. Cats from 162 urban and rural feral cat colonies were sampled over 3 years. Of 766 cats from which nasal swabs were obtained, Cryptococcus spp. were recovered from 95 (12.6%), including 37 C. magnus (4.8%), 16 C. albidus (2.0%), 15 C. carnescens (1.9%), 12 C. neoformans (1.6%), as well as C. oeirensis (n = 3), C. victoriae (n = 3), C. albidosimilis (n = 2), Filobasidium globisporum (n = 2), C. adeliensis (n = 1), C. flavescens (n = 1), C. dimnae (n = 1), C. saitoi (n = 1), and C. wieringae (n = 1) with prevalence feral cats may carry C. neoformans and other Cryptococcus species in their sinonasal cavity. Genotyping of the specific cryptococcal isolates provides a better understanding of the epidemiology of these yeasts. © The Author 2014. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  2. Microbial lipid production by oleaginous yeast Cryptococcus sp. in the batch cultures using corncob hydrolysate as carbon source

    International Nuclear Information System (INIS)

    Chang, Yi-Huang; Chang, Ku-Shang; Lee, Ching-Fu; Hsu, Chuan-Liang; Huang, Cheng-Wei; Jang, Hung-Der

    2015-01-01

    To realize the feasibility of biodiesel production from high-lipid cell culture, microbial lipid production by the oleaginous yeasts was studied using glucose and sucrose as carbon source. Among the tested strains, Cryptococcus sp. SM5S05 accumulated the highest levels of intracellular lipids. The crude lipid contents of Cryptococcus sp. cultured in yeast malt agar reached 30% on a dry weight basis. The accumulation of lipids strongly depended on carbon/nitrogen ratio and nitrogen concentration. The highest content of lipids, measured at a carbon/nitrogen ratio of 60–90 and at a nitrogen concentration of 0.2%, was 60–57% lipids in the dry biomass. Batch cultures using corncob hydrolysate demonstrated that there was minimal inhibitory effect with a reducing sugar concentration of 60 g l −1 or higher. Batch cultures of Cryptococcus sp. SM5S05 in the corncob hydrolysate medium with 60 g l −1 glucose resulted in a dry biomass, lipid yields, and content of 12.6 g l −1 , 7.6 g l −1 , and 60.2%, respectively. The lipids contained mainly long-chain saturated and unsaturated fatty acids with 16 and 18 carbon atoms. The fatty acid profile of Cryptococcus oils was quite similar to that of conventional vegetable oil. The cost of lipid production could be further reduced with corncob hydrolysate being utilized as the raw material for the oleaginous yeast. The results showed that the microbial lipid from Cryptococcus sp. was a potential alternative resource for biodiesel production. - Highlights: • Microbial oil production from oleaginous yeast Cryptococcus sp. was studied. • Accumulation of lipid strongly depended on C/N ratio and nitrogen concentration. • Cultures in hydrolysate medium with 60 g/l glucose resulted in maximum lipid yields. • Maximal lipid content in the Cryptococcus sp. were 60.2% on dried weight basis

  3. Cryptococcus neoformans Is Internalized by Receptor-Mediated or ‘Triggered’ Phagocytosis, Dependent on Actin Recruitment

    Science.gov (United States)

    Guerra, Caroline Rezende; Seabra, Sergio Henrique; de Souza, Wanderley; Rozental, Sonia

    2014-01-01

    Cryptococcosis by the encapsulated yeast Cryptococcus neoformans affects mostly immunocompromised individuals and is a frequent neurological complication in AIDS patients. Recent studies support the idea that intracellular survival of Cryptococcus yeast cells is important for the pathogenesis of cryptococcosis. However, the initial steps of Cryptococcus internalization by host cells remain poorly understood. Here, we investigate the mechanism of Cryptococcus neoformans phagocytosis by peritoneal macrophages using confocal and electron microscopy techniques, as well as flow cytometry quantification, evaluating the importance of fungal capsule production and of host cell cytoskeletal elements for fungal phagocytosis. Electron microscopy analyses revealed that capsular and acapsular strains of C. neoformans are internalized by macrophages via both ‘zipper’ (receptor-mediated) and ‘trigger’ (membrane ruffle-dependent) phagocytosis mechanisms. Actin filaments surrounded phagosomes of capsular and acapsular yeasts, and the actin depolymerizing drugs cytochalasin D and latrunculin B inhibited yeast internalization and actin recruitment to the phagosome area. In contrast, nocodazole and paclitaxel, inhibitors of microtubule dynamics decreased internalization but did not prevent actin recruitment to the site of phagocytosis. Our results show that different uptake mechanisms, dependent on both actin and tubulin dynamics occur during yeast internalization by macrophages, and that capsule production does not affect the mode of Cryptococcus uptake by host cells. PMID:24586631

  4. Genome sequence of a microbial lipid producing fungus Cryptococcus albidus NT2002.

    Science.gov (United States)

    Yong, Xiaoyu; Yan, Zhiying; Xu, Lin; Zhou, Jun; Wu, Xiayuan; Wu, Yuandong; Li, Yang; Chen, Zugeng; Zhou, Hua; Wei, Ping; Jia, Honghua

    2016-04-10

    Cryptococcus albidus NT2002, isolated from the soil in Xinjiang, China, appeared to have the ability to accumulate microbial lipid by utilizing various carbon sources. The predominant properties make it as a potential bio-platform for biodiesel production. Here, we report the complete genome sequence of C. albidus NT2002, which might provide a basis for further elucidation of the genetic background of this promising strain for developing metabolic engineering strategies to produce biodiesel in a green and sustainable manner. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. The Role of Ceramide Synthases in the Pathogenicity of Cryptococcus neoformans.

    Science.gov (United States)

    Munshi, Mansa A; Gardin, Justin M; Singh, Ashutosh; Luberto, Chiara; Rieger, Robert; Bouklas, Tejas; Fries, Bettina C; Del Poeta, Maurizio

    2018-02-06

    Cryptococcus neoformans (C. neoformans) is estimated to cause about 220,000 new cases every year in patients with AIDS, despite advances in antifungal treatments. C. neoformans possesses a remarkable ability to disseminate through an immunocompromised host, making treatment difficult. Here, we examine the mechanism of survival of C. neoformans under varying host conditions and find a role for ceramide synthase in C. neoformans virulence. This study also provides a detailed lipidomics resource for the fungal lipid research community in addition to discovering a potential target for antifungal therapy. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  6. Sterol composition of Cryptococcus neoformans in the presence and absence of fluconazole.

    OpenAIRE

    Ghannoum, M A; Spellberg, B J; Ibrahim, A S; Ritchie, J A; Currie, B; Spitzer, E D; Edwards, J E; Casadevall, A

    1994-01-01

    Analysis of the sterol compositions of 13 clinical isolates of the pathogenic yeast Cryptococcus neoformans obtained from five patients with recurring cryptococcal meningitis showed that, unlike Candida albicans, the major sterols synthesized by this yeast were obtusifoliol (range, 21.1 to 68.2%) and ergosterol (range, 0.0 to 46.5%). There was considerable variation in the sterol contents among the 13 isolates, with total sterol contents ranging from 0.31 to 5.9% of dry weight. The isolates f...

  7. Endogenous Cryptococcus neoformans endophthalmitis with subretinal abscess in a HIV-infected man.

    Science.gov (United States)

    Joseph, Joveeta; Sharma, Savitri; Narayanan, Raja

    2018-07-01

    To report a rare case of Cryptococcus neoformans endogenous endophthalmitis with subretinal abscess in a 36-year-old HIV-positive man, referred with progressive blurred vision in his right eye for the last 6 months. Vitreous biopsy followed by intravitreal ganciclovir did not result in significant improvement. Microbiology revealed the presence of C. neoformans, and intravitreal amphotericin B was then administered. The patient was treated aggressively with systemic and intravitreal antifungals but had a poor visual and anatomical outcome. A high degree of clinical suspicion combined with microbiological evaluation helped to arrive at an appropriate diagnosis.

  8. Contemplating the murine test tube: lessons from natural killer cells and Cryptococcus neoformans.

    Science.gov (United States)

    Marr, Kaleb J; Jones, Gareth J; Mody, Christopher H

    2006-06-01

    Murine experimentation has provided many useful tools, including the ability to knockout or over-express genes and to perform experiments that are limited by ethical considerations. Over the past century, mice have imparted valuable insights into the biology of many systems, including human immunity. However, although there are many similarities between the immune response of humans and mice, there are also many differences; none is more prominent than when examining natural killer cell biology. These differences include tissue distribution, effector molecules, receptor repertoire, and cytokine responses, all of which have important implications when extrapolating the studies to the human immune responses to Cryptococcus neoformans.

  9. The Genome of the Basidiomycetous Yeast and Human Pathogen Cryptococcus neoformans

    Science.gov (United States)

    Loftus, Brendan J.; Fung, Eula; Roncaglia, Paola; Rowley, Don; Amedeo, Paolo; Bruno, Dan; Vamathevan, Jessica; Miranda, Molly; Anderson, Iain J.; Fraser, James A.; Allen, Jonathan E.; Bosdet, Ian E.; Brent, Michael R.; Chiu, Readman; Doering, Tamara L.; Donlin, Maureen J.; D’Souza, Cletus A.; Fox, Deborah S.; Grinberg, Viktoriya; Fu, Jianmin; Fukushima, Marilyn; Haas, Brian J.; Huang, James C.; Janbon, Guilhem; Jones, Steven J. M.; Koo, Hean L.; Krzywinski, Martin I.; Kwon-Chung, June K.; Lengeler, Klaus B.; Maiti, Rama; Marra, Marco A.; Marra, Robert E.; Mathewson, Carrie A.; Mitchell, Thomas G.; Pertea, Mihaela; Riggs, Florenta R.; Salzberg, Steven L.; Schein, Jacqueline E.; Shvartsbeyn, Alla; Shin, Heesun; Shumway, Martin; Specht, Charles A.; Suh, Bernard B.; Tenney, Aaron; Utterback, Terry R.; Wickes, Brian L.; Wortman, Jennifer R.; Wye, Natasja H.; Kronstad, James W.; Lodge, Jennifer K.; Heitman, Joseph; Davis, Ronald W.; Fraser, Claire M.; Hyman, Richard W.

    2012-01-01

    Cryptococcus neoformans is a basidiomycetous yeast ubiquitous in the environment, a model for fungal pathogenesis, and an opportunistic human pathogen of global importance. We have sequenced its ~20-megabase genome, which contains ~6500 intron-rich gene structures and encodes a transcriptome abundant in alternatively spliced and antisense messages. The genome is rich in transposons, many of which cluster at candidate centromeric regions. The presence of these transposons may drive karyotype instability and phenotypic variation. C. neoformans encodes unique genes that may contribute to its unusual virulence properties, and comparison of two phenotypically distinct strains reveals variation in gene content in addition to sequence polymorphisms between the genomes. PMID:15653466

  10. Evidence that the human pathogenic fungus Cryptococcus neoformans var. grubii may have evolved in Africa.

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    Anastasia P Litvintseva

    2011-05-01

    Full Text Available Most of the species of fungi that cause disease in mammals, including Cryptococcus neoformans var. grubii (serotype A, are exogenous and non-contagious. Cryptococcus neoformans var. grubii is associated worldwide with avian and arboreal habitats. This airborne, opportunistic pathogen is profoundly neurotropic and the leading cause of fungal meningitis. Patients with HIV/AIDS have been ravaged by cryptococcosis--an estimated one million new cases occur each year, and mortality approaches 50%. Using phylogenetic and population genetic analyses, we present evidence that C. neoformans var. grubii may have evolved from a diverse population in southern Africa. Our ecological studies support the hypothesis that a few of these strains acquired a new environmental reservoir, the excreta of feral pigeons (Columba livia, and were globally dispersed by the migration of birds and humans. This investigation also discovered a novel arboreal reservoir for highly diverse strains of C. neoformans var. grubii that are restricted to southern Africa, the mopane tree (Colophospermum mopane. This finding may have significant public health implications because these primal strains have optimal potential for evolution and because mopane trees contribute to the local economy as a source of timber, folkloric remedies and the edible mopane worm.

  11. Antifungal Activity of the Volatiles of High Potency Cannabis sativa L. Against Cryptococcus neoformans

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    Amira S. Wanas

    2016-03-01

    Full Text Available The n-hexane extracted volatile fraction of high potency Cannabis sativa L (Cannabaceae . was assessed in vitro for antifungal, antibacterial and antileishmanial activities. The oil exhibited selective albeit modest, antifungal activity against Cryptococcus neoformans with an IC 50 value of 33.1 µg/mL. Biologically-guided fractionation of the volatile fraction resulted in the isolation of three major compounds (1-3 using various chromatographic techniques. The chemical structures of the isolated compounds were identified as α-humulene (1, b -caryophyllene (2 and caryophyllene oxide (3 using GC/FID, GC/MS, 1D- and 2D-NMR analyses, respectively. Compound 1 showed potent and selective antifungal activity against Cryptococcus neoformans with IC 50 and MIC values of 1.18 m g/mL and 5.0 m g/mL respectively. Whereas compound 2 showed weak activity (IC 50 19.4 µg/mL, while compound 3 was inactive against C. neoformans.

  12. Antifungal Activity of Thapsia villosa Essential Oil against Candida, Cryptococcus, Malassezia, Aspergillus and Dermatophyte Species

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    Eugénia Pinto

    2017-09-01

    Full Text Available The composition of the essential oil (EO of Thapsia villosa (Apiaceae, isolated by hydrodistillation from the plant’s aerial parts, was analysed by GC and GC-MS. Antifungal activity of the EO and its main components, limonene (57.5% and methyleugenol (35.9%, were evaluated against clinically relevant yeasts (Candida spp., Cryptococcus neoformans and Malassezia furfur and moulds (Aspergillus spp. and dermatophytes. Minimum inhibitory concentrations (MICs were measured according to the broth macrodilution protocols by Clinical and Laboratory Standards Institute (CLSI. The EO, limonene and methyleugenol displayed low MIC and MFC (minimum fungicidal concentration values against Candida spp., Cryptococcus neoformans, dermatophytes, and Aspergillus spp. Regarding Candida species, an inhibition of yeast–mycelium transition was demonstrated at sub-inhibitory concentrations of the EO (MIC/128; 0.01 μL/mL and their major compounds in Candida albicans. Fluconazole does not show this activity, and the combination with low concentrations of EO could associate a supplementary target for the antifungal activity. The association of fluconazole with T. villosa oil does not show antagonism, but the combination limonene/fluconazole displays synergism. The fungistatic and fungicidal activities revealed by T. villosa EO and its main compounds, associated with their low haemolytic activity, confirm their potential antimicrobial interest against fungal species often associated with human mycoses.

  13. Cryptosporidium,Giardia, Cryptococcus, Pneumocystis genetic variability: cryptic biological species or clonal near-clades?

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    Michel Tibayrenc

    2014-04-01

    Full Text Available An abundant literature dealing with the population genetics and taxonomy of Giardia duodenalis, Cryptosporidium spp., Pneumocystis spp., and Cryptococcus spp., pathogens of high medical and veterinary relevance, has been produced in recent years. We have analyzed these data in the light of new population genetic concepts dealing with predominant clonal evolution (PCE recently proposed by us. In spite of the considerable phylogenetic diversity that exists among these pathogens, we have found striking similarities among them. The two main PCE features described by us, namely highly significant linkage disequilibrium and near-clading (stable phylogenetic clustering clouded by occasional recombination, are clearly observed in Cryptococcus and Giardia, and more limited indication of them is also present in Cryptosporidium and Pneumocystis. Moreover, in several cases, these features still obtain when the near-clades that subdivide the species are analyzed separately ("Russian doll pattern". Lastly, several sets of data undermine the notion that certain microbes form clonal lineages simply owing to a lack of opportunity to outcross due to low transmission rates leading to lack of multiclonal infections ("starving sex hypothesis". We propose that the divergent taxonomic and population genetic inferences advanced by various authors about these pathogens may not correspond to true evolutionary differences and could be, rather, the reflection of idiosyncratic practices among compartmentalized scientific communities. The PCE model provides an opportunity to revise the taxonomy and applied research dealing with these pathogens and others, such as viruses, bacteria, parasitic protozoa, and fungi.

  14. Multilocus sequence typing analysis reveals that Cryptococcus neoformans var. neoformans is a recombinant population.

    Science.gov (United States)

    Cogliati, Massimo; Zani, Alberto; Rickerts, Volker; McCormick, Ilka; Desnos-Ollivier, Marie; Velegraki, Aristea; Escandon, Patricia; Ichikawa, Tomoe; Ikeda, Reiko; Bienvenu, Anne-Lise; Tintelnot, Kathrin; Tore, Okan; Akcaglar, Sevim; Lockhart, Shawn; Tortorano, Anna Maria; Varma, Ashok

    2016-02-01

    Cryptococcus neoformans var. neoformans (serotype D) represents about 30% of the clinical isolates in Europe and is present less frequently in the other continents. It is the prevalent etiological agent in primary cutaneous cryptococcosis as well as in cryptococcal skin lesions of disseminated cryptococcosis. Very little is known about the genotypic diversity of this Cryptococcus subtype. The aim of this study was to investigate the genotypic diversity among a set of clinical and environmental C. neoformans var. neoformans isolates and to evaluate the relationship between genotypes, geographical origin and clinical manifestations. A total of 83 globally collected C. neoformans var. neoformans isolates from Italy, Germany, France, Belgium, Denmark, Greece, Turkey, Thailand, Japan, Colombia, and the USA, recovered from different sources (primary and secondary cutaneous cryptococcosis, disseminated cryptococcosis, the environment, and animals), were included in the study. All isolates were confirmed to belong to genotype VNIV by molecular typing and they were further investigated by MLST analysis. Maximum likelihood phylogenetic as well as network analysis strongly suggested the existence of a recombinant rather than a clonal population structure. Geographical origin and source of isolation were not correlated with a specific MLST genotype. The comparison with a set of outgroup C. neoformans var. grubii isolates provided clear evidence that the two varieties have different population structures. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Different culture media containing methyldopa for melanin production by Cryptococcus species

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    Ralciane de Paula Menezes

    2011-10-01

    Full Text Available INTRODUCTION: Melanin production by species of Cryptococcus is widely used to characterize C. neoformans complex in mycology laboratories. This study aims to test the efficacy of methyldopa from pharmaceutical tablet as a substrate for melanin production, to compare the production of melanin using different agar base added with methyldopa, and to compare the melanin produced in those media with that produced in Niger seed agar and sunflower seed agar by C. neoformans, C. laurentii, and C. albidus. Two isolates of each species, C. neoformans, C. laurentii, and C. albidus, and one of Candida albicans were used to experimentally detect conditions for melanin production. METHODS: The following media were tested: Mueller-Hinton agar (MHA, brain and heart infusion agar (BHIA, blood agar base (BAB, and minimal medium agar (MMA, all added with methyldopa, and the media Niger seed agar (NSA and sunflower seed agar (SSA. RESULTS: All isolates grew in most of the culture media after 24h. Strains planted on media BAB and BHIA showed growth only after 48h. All isolates produced melanin in MMA, MHA, SSA, and NSA media. CONCLUSIONS: Methyldopa in the form pharmaceutical tablet can be used as a substrate for melanin production by Cryptococcus species; minimal medium plus methyldopa was more efficient than the BAB, MHA, and BHIA in the melanin production; and NSA and SSA, followed by MMA added with methyldopa, were more efficient than other media studied for melanin production by all strains studied.

  16. Scalp Lesions in a Pediatric Patient with Hyper IgM Syndrome: Clinical and Histologic Mimicry of Cryptococcus neoformans Infection.

    Science.gov (United States)

    Acker, Karen P; Fetch, Audrey; Schnell, Stephanie A; Hammond, Jennifer; Herrera, Christina; Niedt, George; Ratner, Adam J; Lauren, Christine T

    2018-01-01

    We report a case of cutaneous cryptococcosis due to Cryptococcus neoformans in a pediatric patient with hyper IgM syndrome with scalp lesions that resembled tinea capitis on gross examination and mimicked juvenile xanthogranuloma on histologic examination. This case highlights the importance of considering cutaneous cryptococcosis in patients with hyper IgM syndrome. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Targeting the Cryptococcus neoformans var. grubii Cell Wall Using Lectins: Study of the Carbohydrate-Binding Domain

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    Pamella de Brito Ximenes

    2015-02-01

    Full Text Available Cryptococcus neoformans var. grubii is considered to be the major cause of cryptococcosis in immunosuppressed patients. Understanding cell wall glycoproteins using lectins is of medical interest and can contribute to specific therapy. The aim of this study was to evaluate the carbohydrates on the cell wall of Cryptococcus neoformans var. grubii clinical isolates, using a fluorescein isothiocyanate-lectin binding protocol. Thirty yeast strains stocked in the culture collection were cultivated for 2 days at 30 °C with shaking. Cells were obtained by centrifugation, washed in phosphate-buffered saline, and a suspension of 107 cells/mL was obtained. To determine the binding profile of lectins, concanavalin A (Con A, wheat germ agglutinin (WGA, Ulex europaeus agglutinin I (UEA-I, and peanut agglutinin (PNA conjugated to fluorescein were used. All the tested clinical isolates of Cryptococcus neoformans var. grubii were intensely stained by WGA, moderately stained by Con A, and weakly stained by PNA and UEA-I. Thus, Cryptococcus can be detected in clinical specimens such as blood and cerebrospinal fluid using the fluorescent lectin WGA, which may be considered as an option for detection in cases of suspected cryptococcosis with low laboratory sensitivity. Future applications may be developed using this basic tool.

  18. Targeting the Mitochondrial Respiratory Chain of Cryptococcus through Antifungal Chemosensitization: A Model for Control of Non-Fermentative Pathogens

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    Kathleen L. Chan

    2013-07-01

    Full Text Available Enhanced control of species of Cryptococcus, non-fermentative yeast pathogens, was achieved by chemosensitization through co-application of certain compounds with a conventional antimicrobial drug. The species of Cryptococcus tested showed higher sensitivity to mitochondrial respiratory chain (MRC inhibition compared to species of Candida. This higher sensitivity results from the inability of Cryptococcus to generate cellular energy through fermentation. To heighten disruption of cellular MRC, octyl gallate (OG or 2,3-dihydroxybenzaldehyde (2,3-DHBA, phenolic compounds inhibiting mitochondrial functions, were selected as chemosensitizers to pyraclostrobin (PCS; an inhibitor of complex III of MRC. The cryptococci were more susceptible to the chemosensitization (i.e., PCS + OG or 2,3-DHBA than the Candida with all Cryptococcus strains tested being sensitive to this chemosensitization. Alternatively, only few of the Candida strains showed sensitivity. OG possessed higher chemosensitizing potency than 2,3-DHBA, where the concentration of OG required with the drug to achieve chemosensitizing synergism was much lower than that required of 2,3-DHBA. Bioassays with gene deletion mutants of the model yeast Saccharomyces cerevisiae showed that OG or 2,3-DHBA affect different cellular targets. These assays revealed mitochondrial superoxide dismutase or glutathione homeostasis plays a relatively greater role in fungal tolerance to 2,3-DHBA or OG, respectively. These findings show that application of chemosensitizing compounds that augment MRC debilitation is a promising strategy to antifungal control against yeast pathogens.

  19. Techniques for the detection of pathogenic Cryptococcus species in wood decay substrata and the evaluation of viability in stored samples

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    Christian Alvarez

    2013-02-01

    Full Text Available In this study, we evaluated several techniques for the detection of the yeast form of Cryptococcus in decaying wood and measured the viability of these fungi in environmental samples stored in the laboratory. Samples were collected from a tree known to be positive for Cryptococcus and were each inoculated on 10 Niger seed agar (NSA plates. The conventional technique (CT yielded a greater number of positive samples and indicated a higher fungal density [in colony forming units per gram of wood (CFU.g-1] compared to the humid swab technique (ST. However, the difference in positive and false negative results between the CT-ST was not significant. The threshold of detection for the CT was 0.05.10³ CFU.g-1, while the threshold for the ST was greater than 0.1.10³ CFU-1. No colonies were recovered using the dry swab technique. We also determined the viability of Cryptococcus in wood samples stored for 45 days at 25ºC using the CT and ST and found that samples not only continued to yield a positive response, but also exhibited an increase in CFU.g-1, suggesting that Cryptococcus is able to grow in stored environmental samples. The ST.1, in which samples collected with swabs were immediately plated on NSA medium, was more efficient and less laborious than either the CT or ST and required approximately 10 min to perform; however, additional studies are needed to validate this technique.

  20. Eucalyptus Tree: A Potential Source of Cryptococcus neoformans in Egyptian Environment

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    Mahmoud Elhariri

    2016-01-01

    Full Text Available In Egypt, the River Red Gum (Eucalyptus camaldulensis is a well-known tree and is highly appreciated by the rural and urban dwellers. The role of Eucalyptus trees in the ecology of Cryptococcus neoformans is documented worldwide. The aim of this survey was to show the prevalence of C. neoformans during the flowering season of E. camaldulensis at the Delta region in Egypt. Three hundred and eleven samples out of two hundred Eucalyptus trees, including leaves, flowers, and woody trunks, were collected from four governorates in the Delta region. Thirteen isolates of C. neoformans were recovered from Eucalyptus tree samples (4.2%. Molecular identification of C. neoformans was done by capsular gene specific primer CAP64 and serotype identification was done depending on LAC1 gene. This study represents an update on the ecology of C. neoformans associated with Eucalyptus tree in Egyptian environment.

  1. 3-Bromopyruvate: a novel antifungal agent against the human pathogen Cryptococcus neoformans.

    Science.gov (United States)

    Dyląg, Mariusz; Lis, Paweł; Niedźwiecka, Katarzyna; Ko, Young H; Pedersen, Peter L; Goffeau, Andre; Ułaszewski, Stanisław

    2013-05-03

    We have investigated the antifungal activity of the pyruvic acid analogue: 3-bromopyruvate (3-BP). Growth inhibition by 3-BP of 110 strains of yeast-like and filamentous fungi was tested by standard spot tests or microdilution method. The human pathogen Cryptococcus neoformans exhibited a low Minimal Inhibitory Concentration (MIC) of 0.12-0.15 mM 3-BP. The high toxicity of 3-BP toward C. neoformans correlated with high intracellular accumulation of 3-BP and also with low levels of intracellular ATP and glutathione. Weak cytotoxicity towards mammalian cells and lack of resistance conferred by the PDR (Pleiotropic Drug Resistance) network in the yeast Saccharomyces cerevisiae, are other properties of 3-BP that makes it a novel promising anticryptococcal drug. Copyright © 2013 Elsevier Inc. All rights reserved.

  2. Perfect state of Cryptococcus neoformans, Filobasidiella neoformans, on pigeon manure filtrate agar

    Energy Technology Data Exchange (ETDEWEB)

    Staib, F.

    1981-02-01

    To enable studies of the dependence of Cryptococcus neoformans and its perfect and imperfect states upon bird manure as a habitat of this pathogen, a nutrient medium closely resembling natural conditions was prepared. As sole nutrient, the water soluble ingredients of manure from pigeons (Columbia livia) were used. There was no heat sterilization of the manure filtrate. Using a standard pair of C. neoformans strains for mating, it could be demonstrated that the perfect state of the fungus developed on this so called pigeon manure filtrate agar within 48 h at 26 degrees C. This medium is supposed to help in the elucidation of the epidemiological significance of the perfect and imperfect states of this pathogen.

  3. Paramecium species ingest and kill the cells of the human pathogenic fungus Cryptococcus neoformans.

    Science.gov (United States)

    Frager, Shalom Z; Chrisman, Cara J; Shakked, Rachel; Casadevall, Arturo

    2010-08-01

    A fundamental question in the field of medical mycology is the origin of virulence in those fungal pathogens acquired directly from the environment. In recent years, it was proposed that the virulence of certain environmental animal-pathogenic microbes, such as Cryptococcus neoformans, originated from selection pressures caused by species-specific predation. In this study, we analyzed the interaction of C. neoformans with three Paramecium spp., all of which are ciliated mobile protists. In contrast to the interaction with amoebae, some Paramecium spp. rapidly ingested C. neoformans and killed the fungus. This study establishes yet another type of protist-fungal interaction supporting the notion that animal-pathogenic fungi in the environment are under constant selection by predation.

  4. PENAPISAN KHAMIR SELULOLITIK CRYPTOCOCCUS SP. YANG DIISOLASI DARI TANAH KEBUN BIOLOGI WAMENA, JAYA WIJAYA, PROPINSI PAPUA

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    Atit Kanti

    2007-06-01

    Full Text Available Cryptococcus sp. was isolated from Kebun Biologi Wamena, Papua. The isolate was able to grow in media with carboxymethyl cellulose as a sole carbon source implying that isolate produced 1-3 ? endo-glucanase. To study the effect of glucose and osmotic pressure, 0.1 % glucose and 0.1 % NaCl were amended into the medium containing CMC. Glucose significantly affected cellulolytic activity and biomass synthesis. At the beginning of cell cultivation glucose augmentation appear to slightly inhibit enzyme activity. Sodium chloride also significantly affected cellulolytic activity. Profile of pH varied dependent on cultivation media. Maximum growth of biomass was achieved after glucose addition, indicating that glucose stimulated cell growth.

  5. Antagonistic Activity of Probiotic Organism Against Vibrio cholerae and Cryptococcus neoformans

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    Vidya, R.

    2010-01-01

    Full Text Available The microbes are useful in many ways in the modern world. Probiotics one of them, which refers to, acid adherence bacteria in the intestinal cells, are able to survive at low pH and produce large amount of lactic acid. The present investigation deals with the antagonistic activity of Lactobacillus acidophilus organism against pathogens. The organism was isolated from the curd sample. Identification of bacteria was done by various biochemical testing. The present study revealed that L. acidophilus inhibits Vibrio cholerae more efficiently than Streptococcus pneumoniae and Shigella dysentriae. When L. acidophilus and V. cholerae were grown together, L. acidophilus dominated the growth and competitively inhibited the growth of V. cholerae. L. acidophilus was also found to inhibit Cryptococcus neoformans.

  6. Isolation of Cryptococcus neoformans var. neoformans from bird droppings, fruits and vegetables in Mexico City.

    Science.gov (United States)

    López-Martínez, R; Castañón-Olivares, L R

    1995-01-01

    The presence of Cryptococcus neoformans in various natural sources, such as bird droppings, fruits and vegetables, was investigated. A total of 711 samples were analyzed; C. neoformans var. neoformans was isolated from seven out of 74 bird droppings (9.5%), with parrots as one of the most significant sources. Fruits were positive in 9.5% of the 169 samples studied, specially citrus fruits, particularly grapefruit, in which the highest frequency was found. From the 468 vegetable samples, only 20 were positive (4.2%). It is emphasized that five of the positive vegetables species are autochthonous to Mexico: avocado (Nectandra salicifolia), beet (Beta vulgaris var. quinopodiace), chayote (Sechium edule), stringbean (Cassia sp), and nopal (Opuntia ficus-indica).

  7. Pilzinfektionen des Zentralnervensystems bei immunkompetentem Wirt [Fungal infections of the central nervous system in the immunocompetent host

    NARCIS (Netherlands)

    Tintelnot, K.; de Hoog, G.S.; Haase, G.

    2013-01-01

    The majority of mycoses which lead to mycotic tumors in patients without any predisposing underlying disease are either caused by Cryptococcus gattii and C. neoformans or by dematiaceous fungi which include Cladophialophora bantiana, Ramichloridium mackenziei, Exophiala and Fonsecaea species. The

  8. The Role of Cryptococcus in the Immune System of Pulmonary Cryptococcosis Patients.

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    Jinlin Wang

    Full Text Available To investigate the role of Cryptococcus in the immune system of immunocompetent patients with pulmonary cryptococcosis (PC by analysing the dynamic changes of patients' immune status before and after antifungal therapy.The level of the serum interferon-γ (IFN-γ and interleukin (IL-2, -4, -10 and -12 was measured before and after 6-months of treatment. Peripheral blood samples were obtained from 30 immunocompetent PC patients and 30 age- and gender-matched healthy controls. Peripheral blood mononuclear cells (PBMCs were isolated and incubated with recombinant human IL-12 (rhIL-12 for 48 h. Then the concentrations of IFN-γ and IL-4 in the supernatant were analysed.Baseline serum IFN-γ level was significantly lower in the PC patients as compared with the control group (P < 0.001. The serum IL-2 and IFN-γ of PC patients were significantly increased after appropriate treatments (P < 0.05 and P < 0.001 when compared to their baseline levels. The productions of IFN-γ in the culture supernatant of PBMCs showed no significant difference between the control and PC patients both before and after antifungal treatments. RhIL-12 is a potent stimulus for IFN-γ production. Culture PBMCs collected from PC patients before treatments had a smaller increase of IFN-γ production in the present of rhIL-12 than the control (P < 0.01; PBMCs from PC patients completing 6-months of treatment showed a comparable increase of IFN-γ production by rhIL-12 stimulation to the control group.In apparently immunocompetent patients with PC, a normalization of serum IFN-γ was achieved after recovery from infection. This suggests that Cryptococcus infection per se can suppress the immune system and its elimination contributes to the reestablishment of an immune equilibrium.

  9. Methamphetamine enhances Cryptococcus neoformans pulmonary infection and dissemination to the brain.

    Science.gov (United States)

    Patel, Dhavan; Desai, Gunjan M; Frases, Susana; Cordero, Radames J B; DeLeon-Rodriguez, Carlos M; Eugenin, Eliseo A; Nosanchuk, Joshua D; Martinez, Luis R

    2013-07-30

    Methamphetamine (METH) is a major addictive drug of abuse in the United States and worldwide, and its use is linked to HIV acquisition. The encapsulated fungus Cryptococcus neoformans is the most common cause of fungal meningitis in patients with AIDS. In addition to functioning as a central nervous system stimulant, METH has diverse effects on host immunity. Using a systemic mouse model of infection and in vitro assays in order to critically assess the impact of METH on C. neoformans pathogenesis, we demonstrate that METH stimulates fungal adhesion, glucuronoxylomannan (GXM) release, and biofilm formation in the lungs. Interestingly, structural analysis of the capsular polysaccharide of METH-exposed cryptococci revealed that METH alters the carbohydrate composition of this virulence factor, an event of adaptation to external stimuli that can be advantageous to the fungus during pathogenesis. Additionally, we show that METH promotes C. neoformans dissemination from the respiratory tract into the brain parenchyma. Our findings provide novel evidence of the impact of METH abuse on host homeostasis and increased permissiveness to opportunistic microorganisms. Methamphetamine (METH) is a major health threat to our society, as it adversely changes people's behavior, as well as increases the risk for the acquisition of diverse infectious diseases, particularly those that enter through the respiratory tract or skin. This report investigates the effects of METH use on pulmonary infection by the AIDS-related fungus Cryptococcus neoformans. This drug of abuse stimulates colonization and biofilm formation in the lungs, followed by dissemination of the fungus to the central nervous system. Notably, C. neoformans modifies its capsular polysaccharide after METH exposure, highlighting the fungus's ability to adapt to environmental stimuli, a possible explanation for its pathogenesis. The findings may translate into new knowledge and development of therapeutic and public health

  10. The Role of Amino Acid Permeases and Tryptophan Biosynthesis in Cryptococcus neoformans Survival.

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    João Daniel Santos Fernandes

    Full Text Available Metabolic diversity is an important factor during microbial adaptation to different environments. Among metabolic processes, amino acid biosynthesis has been demonstrated to be relevant for survival for many microbial pathogens, whereas the association between pathogenesis and amino acid uptake and recycling are less well-established. Cryptococcus neoformans is an opportunistic fungal pathogen with many habitats. As a result, it faces frequent metabolic shifts and challenges during its life cycle. Here we studied the C. neoformans tryptophan biosynthetic pathway and found that the pathway is essential. RNAi indicated that interruptions in the biosynthetic pathway render strains inviable. However, auxotroph complementation can be partially achieved by tryptophan uptake when a non preferred nitrogen source and lower growth temperature are applied, suggesting that amino acid permeases may be the target of nitrogen catabolism repression (NCR. We used bioinformatics to search for amino acid permeases in the C. neoformans and found eight potential global permeases (AAP1 to AAP8. The transcriptional profile of them revealed that they are subjected to regulatory mechanisms which are known to respond to nutritional status in other fungi, such as (i quality of nitrogen (Nitrogen Catabolism Repression, NCR and carbon sources (Carbon Catabolism Repression, CCR, (ii amino acid availability in the extracellular environment (SPS-sensing and (iii nutritional deprivation (Global Amino Acid Control, GAAC. This study shows that C. neoformans has fewer amino acid permeases than other model yeasts, and that these proteins may be subjected to complex regulatory mechanisms. Our data suggest that the C. neoformans tryptophan biosynthetic pathway is an excellent pharmacological target. Furthermore, inhibitors of this pathway cause Cryptococcus growth arrest in vitro.

  11. Magnesium Ion Acts as a Signal for Capsule Induction in Cryptococcus neoformans.

    Science.gov (United States)

    Rathore, Sudarshan S; Raman, Thiagarajan; Ramakrishnan, Jayapradha

    2016-01-01

    Cryptococcal meningitis caused by Cryptococcus neoformans, is a common opportunistic neural infection in immunocompromised individuals. Cryptococcus meningitis is associated with fungal burden with larger capsule size in cerebrospinal fluid (CSF). To understand the role of CSF constituents in capsule enlargement, we have evaluated the effect of artificial CSF on capsule induction in comparison with various other capsule inducing media. Two different strains of C. neoformans, an environmental and a clinical isolates were used in the present study. While comparing the various capsule inducing media for the two different strains of C. neoformans, it was observed that the capsule growth was significantly increased when grown in artificial CSF at pH 5.5, temperature 34°C for ATCC C. neoformans and 37°C for Clinical C. neoformans and with an incubation period of 72 h. In addition, artificial CSF supports biofilm formation in C. neoformans. While investigating the individual components of artificial CSF, we found that Mg(2+) ions influence the capsule growth in both environmental and clinical strains of C. neoformans. To confirm our results we studied the expression of four major CAP genes namely, CAP10, CAP59, CAP60, and CAP64 in various capsule inducing media and in different concentrations of Mg(2+) and Ca(2+). Our results on gene expression suggest that, Mg(2+) does have an effect on CAP gene expression, which are important for capsule biosynthesis and virulence. Our findings on the role of Mg(2+) ion as a signal for capsule induction will promote a way to elucidate the control mechanisms for capsule biosynthesis in C. neoformans.

  12. A new F-actin structure in fungi: actin ring formation around the cell nucleus of Cryptococcus neoformans.

    Science.gov (United States)

    Kopecká, Marie; Kawamoto, Susumu; Yamaguchi, Masashi

    2013-04-01

    The F-actin cytoskeleton of Cryptococcus neoformans is known to comprise actin cables, cortical patches and cytokinetic ring. Here, we describe a new F-actin structure in fungi, a perinuclear F-actin collar ring around the cell nucleus, by fluorescent microscopic imaging of rhodamine phalloidin-stained F-actin. Perinuclear F-actin rings form in Cryptococcus neoformans treated with the microtubule inhibitor Nocodazole or with the drug solvent dimethyl sulfoxide (DMSO) or grown in yeast extract peptone dextrose (YEPD) medium, but they are absent in cells treated with Latrunculin A. Perinuclear F-actin rings may function as 'funicular cabin' for the cell nucleus, and actin cables as intracellular 'funicular' suspending nucleus in the central position in the cell and moving nucleus along the polarity axis along actin cables.

  13. Cryptococcus neoformans en el contenido gástrico de un paciente con SIDA Cryptococcus neoformans in the gastric contents of an AIDS patient

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    S. Garro

    2006-12-01

    Full Text Available Se comunica la observación microscópica y el aislamiento de Cryptococcus neoformans de contenido gástrico de un paciente con SIDA, obtenido por aspiración con sonda nasogástrica y enviado para su estudio parasitológico. En el examen en fresco del concentrado del material, llamaron la atención escasas levaduras redondas. El agregado de tinta china reveló la cápsula característica de C. neoformans. El cultivo de la muestra en agar extracto de semillas de girasol incubado a 37 °C durante 7 días permitió el aislamiento de colonias pardas de C. neoformans. Los hemocultivos para hongos fueron negativos y ante la imposibilidad de obtener LCR, por negativa del paciente, se determinó el antígeno polisacarídico capsular de C. neoformans en sangre, que fue positivo hasta la dilución 1:100. Luego del hallazgo de C. neoformans en el contenido gástrico y del resultado positivo de la antigenemia, el paciente -sospechado inicialmente como portador de diarrea por Cryptosporidium sp.-, fue medicado con fluconazol por vía oral, a razón de 800 mg/día, tras negarse a recibir medicación endovenosa. La presente comunicación da cuenta del hallazgo de C. neoformans en un material clínico donde su presencia es infrecuente y destaca su valor en el diagnóstico de la criptococosis.The microscopic observation and isolation of Cryptococcus neoformans from the gastric contents of an AIDS patient, obtained by aspiration with a nasogastric catheter and parasitologically studied, is communicated. Because of the limited number of round yeasts visualized by wet mount of the sample concentrate, India ink was added: the typical capsules of C. neoformans were then observed. Dark brown colonies of C. neoformans were isolated from the clinical sample cultured on sunflower-seed-extract agar, incubated at 37 °C for 7 days. Bloodcultures for fungi were negative; it was impossible to obtain CSF due to the patient&'s refusal, then the capsular polysaccharide antigen

  14. [A pediatric case of HIV who diagnosed by virtue of disseminated cryptococcus infection].

    Science.gov (United States)

    Acar, Manolya; Sütçü, Murat; Aktürk, Hacer; Hançerli Törün, Selda; Karagöz, Nurinisa; Beka, Hayati; Yekeler, Ensar; Ağaçfidan, Ali; Salman, Nuran; Somer, Ayper

    2016-07-01

    Cryptococcus neoformans is an important opportunistic pathogen that causes serious mortality and morbidity in AIDS patients. Although its incidence has decreased with proper antiretroviral treatment (ART), it is still a major concern in areas with low socioeconomic HIV endemic countries with poor sources of therapy. In our country, pediatric HIV infection and so, HIV-related opportunistic infections are very rare. In order to pay attention to this unusual collaboration; herein, we presented a pediatric case who was diagnosed with HIV and disseminated cryptococcus infection concomitantly. A 6.5-year-old previously healthy girl has admitted to our hospital with the complaints of prolonged fever, cough and hemoptysis. On her physical examination she had oral candidiasis, generalized lymphadenopathy and hepatosplenomegaly. Laboratory findings were as follows; white blood cell count: 3170 µL (neutrophil: 2720 µL, lymphocyte: 366 µL), hemoglobin level: 7.8 gr/dl, hematocrit: 25.5% platelets: 170.000 µL, CRP: 15.2 mg/L and serum IgG level: 1865 mg/dl. Her anti-HIV test yielde,d positive result and confirmed by Western blot assay, together with a high viral load (HIV-RNA: 3.442.000 copies/ml). She was started ART (lamivudine, zidovudine and lopinavir/ritonavir combination) with the diagnosis of stage 3 HIV infection (AIDS). Posteroanterior chest radiograph showed mediastinal extension and nodular parenchyma. Since the patient was suspected to have pulmonary tuberculosis based on the clinical and radiological findings, empirical antituberculosis therapy was started. Because of the insistance of fever, three different blood specimens, bone marrow and gastric aspirates were collected for culture, in which all of them yielded C.neoformans growth. She was then diagnosed as disseminated cryptococcosis and treated with liposomal amphotericin B and fluconazole successfully. Although pediatric HIV infection is usually diagnosed secondary to maternal disease, it can rarely be

  15. Secondary cell wall formation in Cryptococcus neoformans as a rescue mechanism against acid-induced autolysis.

    Science.gov (United States)

    Farkas, Vladimír; Takeo, Kanji; Maceková, Danka; Ohkusu, Misako; Yoshida, Soichi; Sipiczki, Matthias

    2009-03-01

    Growth of the opportunistic yeast pathogen Cryptococcus neoformans in a synthetic medium containing yeast nitrogen base and 1.0-3.0% glucose is accompanied by spontaneous acidification of the medium, with its pH decreasing from the initial 5.5 to around 2.5 in the stationary phase. During the transition from the late exponential to the stationary phase of growth, many cells died as a consequence of autolytic erosion of their cell walls. Simultaneously, there was an increase in an ecto-glucanase active towards beta-1,3-glucan and having a pH optimum between pH 3.0 and 3.5. As a response to cell wall degradation, some cells developed an unusual survival strategy by forming 'secondary' cell walls underneath the original ones. Electron microscopy revealed that the secondary cell walls were thicker than the primary ones, exposing bundles of polysaccharide microfibrils only partially masked by an amorphous cell wall matrix on their surfaces. The cells bearing secondary cell walls had a three to five times higher content of the alkali-insoluble cell wall polysaccharides glucan and chitin, and their chitin/glucan ratio was about twofold higher than in cells from the logarithmic phase of growth. The cell lysis and the formation of the secondary cell walls could be suppressed by buffering the growth medium between pH 4.5 and 6.5.

  16. Iron regulation of the major virulence factors in the AIDS-associated pathogen Cryptococcus neoformans.

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    Won Hee Jung

    2006-11-01

    Full Text Available Iron overload is known to exacerbate many infectious diseases, and conversely, iron withholding is an important defense strategy for mammalian hosts. Iron is a critical cue for Cryptococcus neoformans because the fungus senses iron to regulate elaboration of the polysaccharide capsule that is the major virulence factor during infection. Excess iron exacerbates experimental cryptococcosis and the prevalence of this disease in Sub-Saharan Africa has been associated with nutritional and genetic aspects of iron loading in the background of the HIV/AIDS epidemic. We demonstrate that the iron-responsive transcription factor Cir1 in Cr. neoformans controls the regulon of genes for iron acquisition such that cir1 mutants are "blind" to changes in external iron levels. Cir1 also controls the known major virulence factors of the pathogen including the capsule, the formation of the anti-oxidant melanin in the cell wall, and the ability to grow at host body temperature. Thus, the fungus is remarkably tuned to perceive iron as part of the disease process, as confirmed by the avirulence of the cir1 mutant; this characteristic of the pathogen may provide opportunities for antifungal treatment.

  17. Bioconversion of mixed volatile fatty acids into microbial lipids by Cryptococcus curvatus ATCC 20509.

    Science.gov (United States)

    Liu, Jia; Yuan, Ming; Liu, Jia-Nan; Huang, Xiang-Feng

    2017-10-01

    The oleaginous yeast Cryptococcus curvatus ATCC 20509 can use 5-40g/L of acetic, propionic, or butyric acid as sole carbon source to produce lipids. High concentrations (30g/L) of mixed volatile fatty acids (VFAs) were used to cultivate C. curvatus to explore the effects of different ratios of mixed VFAs on lipid production and composition. When mixed VFAs (VFA ratio was 15:5:10) were used as carbon sources, the highest cell mass and lipid concentration were 8.68g/L and 4.93g/L, respectively, which were significantly higher than those when 30g/L of acetic acid was used as sole carbon source. The highest content and yield of odd-numbered fatty acids were 45.1% (VFA ratio was 0:15:15) and 1.62g/L (VFA ratio was 5:15:10), respectively. These results indicate that adjusting the composition ratios of mixed VFAs effectively improves microbial lipid synthesis and the yield of odd-numbered fatty acids. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. A vanillin derivative causes mitochondrial dysfunction and triggers oxidative stress in Cryptococcus neoformans.

    Science.gov (United States)

    Kim, Jin Hyo; Lee, Han-Ok; Cho, Yong-Joon; Kim, Jeongmi; Chun, Jongsik; Choi, Jaehyuk; Lee, Younghoon; Jung, Won Hee

    2014-01-01

    Vanillin is a well-known food and cosmetic additive and has antioxidant and antimutagenic properties. It has also been suggested to have antifungal activity against major human pathogenic fungi, although it is not very effective. In this study, the antifungal activities of vanillin and 33 vanillin derivatives against the human fungal pathogen Cryptococcus neoformans, the main pathogen of cryptococcal meningitis in immunocompromised patients, were investigated. We found a structural correlation between the vanillin derivatives and antifungal activity, showing that the hydroxyl or alkoxy group is more advantageous than the halogenated or nitrated group in benzaldehyde. Among the vanillin derivatives with a hydroxyl or alkoxy group, o-vanillin and o-ethyl vanillin showed the highest antifungal activity against C. neoformans. o-Vanillin was further studied to understand the mechanism of antifungal action. We compared the transcriptome of C. neoformans cells untreated or treated with o-vanillin by using RNA sequencing and found that the compound caused mitochondrial dysfunction and triggered oxidative stress. These antifungal mechanisms of o-vanillin were experimentally confirmed by the significantly reduced growth of the mutants lacking the genes involved in mitochondrial functions and oxidative stress response.

  19. Cryptococcus spp. isolation from excreta of pigeons (Columba livia) in and around Monterrey, Mexico.

    Science.gov (United States)

    Canónico-González, Yolanda; Adame-Rodríguez, Juan Manuel; Mercado-Hernández, Roberto; Aréchiga-Carvajal, Elva Teresa

    2013-01-01

    The presence of Cryptococcus spp. has been reported in Mexico's capital city; however, to our knowledge there are no reports of its presence in the state of Nuevo León located in northeast Mexico. This is presumed to be because the hot and dry climate in this region does not favor cryptococcal proliferation. This study confirmed the presence of C. neoformans and C. albidus in 20% (10/50) of randomly selected fecal samples of pigeons (Columba livia) in the Monterrey metropolitan area. The presence of this yeast in the state of Nuevo León is proof of its adaptation to the typically hot climate of the area and is consistent with recent reviews of cryptococcosis cases in several local hospitals. The two species were identified and characterized through microbiological tests and molecular identification by DNA extraction and PCR amplification of highly conserved 18S ribosomal DNA using ITS1 and ITS2 as target regions. The PCR products were sequenced and compared with those reported in GenBank.

  20. Optimization of lipids production by Cryptococcus laurentii 11 using cheese whey with molasses

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    Rodrigo Fernandes Castanha

    2014-06-01

    Full Text Available This study aimed the optimization of culture condition and composition for production of Cryptococcus laurentii 11 biomass and lipids in cheese whey medium supplemented with sugarcane molasses. The optimization of pH, fermentation time, and molasses concentration according to a full factorial statistical experimental design was followed by a Plackett-Burman experimental design, which was used to determine whether the supplementation of the culture medium by yeast extract and inorganic salts could provide a further enhancement of lipids production. The following conditions and composition of the culture medium were found to optimize biomass and lipids production: 360 h fermentation, 6.5 pH and supplementation of (g L-1: 50 molasses, 0.5 yeast extract, 4 KH2PO4, 1 Na2HPO4, 0.75 MgSO4•7H2O and 0.002 ZnSO4•H2O. Additional supplementation with inorganic salts and yeast extract was essential to optimize the production, in terms of product concentration and productivity, of neutral lipids by C. laurentii 11. Under this optimized condition, the production of total lipids increased by 133% in relation to control experiment (from 1.27 to 2.96 g L-1. The total lipids indicated a predominant (86% presence of neutral lipids with high content of 16- and 18- carbon-chain saturated and monosaturated fatty acids. This class of lipids is considered especially suitable for the production of biodiesel.

  1. Differences in Sirtuin Regulation in Response to Calorie Restriction in Cryptococcus neoformans.

    Science.gov (United States)

    Bouklas, Tejas; Masone, Lindsey; Fries, Bettina C

    2018-02-18

    Cryptococcus neoformans successfully replicates in low glucose in infected patients. In the serotype A strain, H99, growth in this condition prolongs lifespan regulated by SIR2, and can be modulated with SIR2-specific drugs. Previous studies show that lifespan modulation of a cryptococcal population affects its sensitivity to antifungals, and survival in an infection model. Sirtuins and their role in longevity are conserved among fungi; however, the effect of glucose starvation is not confirmed even in Saccharomyces cerevisiae. Lifespan analysis of C. neoformans strains in low glucose showed that 37.5% exhibited pro-longevity, and lifespan of a serotype D strain, RC2, was shortened. Transcriptome comparison of H99 and RC2 under calorie restriction demonstrated differences, confirmed by real-time PCR showing that SIR2 , TOR1 , SCH9 , and PKA1 expression correlated with lifespan response to calorie restriction. As expected, RC2 -sir2 Δ cells exhibited a shortened lifespan, which was reconstituted. However, shortened lifespan from calorie restriction was independent of SIR2 . In contrast to H99 but consistent with altered SIR2 regulation, SIR2 -specific drugs did not affect outcome of RC2 infection. These data suggest that SIR2 regulation and response to calorie restriction varies in C. neoformans, which should be considered when Sirtuins are investigated as potential therapy targets for fungal infections.

  2. Cryptococcus neoformans sexual reproduction is controlled by a quorum sensing peptide.

    Science.gov (United States)

    Tian, Xiuyun; He, Guang-Jun; Hu, Pengjie; Chen, Lei; Tao, Changyu; Cui, Ying-Lu; Shen, Lan; Ke, Weixin; Xu, Haijiao; Zhao, Youbao; Xu, Qijiang; Bai, Fengyan; Wu, Bian; Yang, Ence; Lin, Xiaorong; Wang, Linqi

    2018-06-01

    Bacterial quorum sensing is a well-characterized communication system that governs a large variety of collective behaviours. By comparison, quorum sensing regulation in eukaryotic microbes remains poorly understood, especially its functional role in eukaryote-specific behaviours, such as sexual reproduction. Cryptococcus neoformans is a prevalent fungal pathogen that has two defined sexual cycles (bisexual and unisexual) and is a model organism for studying sexual reproduction in fungi. Here, we show that the quorum sensing peptide Qsp1 serves as an important signalling molecule for both forms of sexual reproduction. Qsp1 orchestrates various differentiation and molecular processes, including meiosis, the hallmark of sexual reproduction. It activates bisexual mating, at least in part through the control of pheromone, a signal necessary for bisexual activation. Notably, Qsp1 also plays a major role in the intercellular regulation of unisexual initiation and coordination, in which pheromone is not strictly required. Through a multi-layered genetic screening approach, we identified the atypical zinc finger regulator Cqs2 as an important component of the Qsp1 signalling cascade during both bisexual and unisexual reproduction. The absence of Cqs2 eliminates the Qsp1-stimulated mating response. Together, these findings extend the range of behaviours governed by quorum sensing to sexual development and meiosis.

  3. Genome-Wide Transcription Study of Cryptococcus neoformans H99 Clinical Strain versus Environmental Strains.

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    Elaheh Movahed

    Full Text Available The infection of Cryptococcus neoformans is acquired through the inhalation of desiccated yeast cells and basidiospores originated from the environment, particularly from bird's droppings and decaying wood. Three environmental strains of C. neoformans originated from bird droppings (H4, S48B and S68B and C. neoformans reference clinical strain (H99 were used for intranasal infection in C57BL/6 mice. We showed that the H99 strain demonstrated higher virulence compared to H4, S48B and S68B strains. To examine if gene expression contributed to the different degree of virulence among these strains, a genome-wide microarray study was performed to inspect the transcriptomic profiles of all four strains. Our results revealed that out of 7,419 genes (22,257 probes examined, 65 genes were significantly up-or down-regulated in H99 versus H4, S48B and S68B strains. The up-regulated genes in H99 strain include Hydroxymethylglutaryl-CoA synthase (MVA1, Mitochondrial matrix factor 1 (MMF1, Bud-site-selection protein 8 (BUD8, High affinity glucose transporter 3 (SNF3 and Rho GTPase-activating protein 2 (RGA2. Pathway annotation using DAVID bioinformatics resource showed that metal ion binding and sugar transmembrane transporter activity pathways were highly expressed in the H99 strain. We suggest that the genes and pathways identified may possibly play crucial roles in the fungal pathogenesis.

  4. Cryptococcus neoformans-derived microvesicles enhance the pathogenesis of fungal brain infection.

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    Sheng-He Huang

    Full Text Available Cryptococcal meningoencephalitis is the most common fungal disease in the central nervous system. The mechanisms by which Cryptococcus neoformans invades the brain are largely unknown. In this study, we found that C. neoformans-derived microvesicles (CnMVs can enhance the traversal of the blood-brain barrier (BBB by C. neoformans invitro. The immunofluorescence imaging demonstrates that CnMVs can fuse with human brain microvascular endothelial cells (HBMECs, the constituents of the BBB. This activity is presumably due to the ability of the CnMVs to activate HBMEC membrane rafts and induce cell fusogenic activity. CnMVs also enhanced C. neoformans infection of the brain, found in both infected brains and cerebrospinal fluid. In infected mouse brains, CnMVs are distributed inside and around C. neoformans-induced cystic lesions. GFAP (glial fibrillary acidic protein-positive astrocytes were found surrounding the cystic lesions, overlapping with the 14-3-3-GFP (14-3-3-green fluorescence protein fusion signals. Substantial changes could be observed in areas that have a high density of CnMV staining. This is the first demonstration that C. neoformans-derived microvesicles can facilitate cryptococcal traversal across the BBB and accumulate at lesion sites of C. neoformans-infected brains. Results of this study suggested that CnMVs play an important role in the pathogenesis of cryptococcal meningoencephalitis.

  5. Isolation of Cryptococcus neoformans and other opportunistic fungi from pigeon droppings.

    Science.gov (United States)

    Soltani, Maryam; Bayat, Mansour; Hashemi, Seyed J; Zia, Mohammadali; Pestechian, Nader

    2013-01-01

    Invasive fungal infections cause considerable morbidity and mortality in immunocompromised hosts. Pigeon droppings could especially be a potential carrier in the spread of pathogenic yeasts and mold fungi into the environment. The objective of this study was to isolation of Cryptococcus neoformans and other opportunistic fungi from pigeon droppings. One hundred twenty samples of pigeon droppings were suspended 1:10 in saline solution and then cultured. Identification of C. neoformans was performed on bird seed agar, presence of a capsule on India ink preparation, urease production on urea agar medium and RapID yeast plus system. The identification of candida species was based on micro-morphological analysis on corn meal-Tween 80 agar, RapID yeast plus system and growth in CHROMagar candida. The identification of other fungi was based on macromorphologic, microscopic, biochemical and physiological characteristics. The highest frequency of yeasts and mold fungi were observed in Candida albicans 6.6% and Penicillium spp. 25%. The frequency rate of C. neoformans isolation was 2.5%. Several types of fungi are present in pigeon droppings that can spread in environment and transmit to children and elderly as well as immunocompromised patients who are at increased risk of contracting opportunistic diseases.

  6. Isolation of Cryptococcus neoformans and other opportunistic fungi from pigeon droppings

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    Maryam Soltani

    2013-01-01

    Full Text Available Background: Invasive fungal infections cause considerable morbidity and mortality in immunocompromised hosts. Pigeon droppings could especially be a potential carrier in the spread of pathogenic yeasts and mold fungi into the environment. The objective of this study was to isolation of Cryptococcus neoformans and other opportunistic fungi from pigeon droppings. Materials and Methods: One hundred twenty samples of pigeon droppings were suspended 1:10 in saline solution and then cultured. Identification of C. neoformans was performed on bird seed agar, presence of a capsule on India ink preparation, urease production on urea agar medium and RapID yeast plus system. The identification of candida species was based on micro-morphological analysis on corn meal-Tween 80 agar, RapID yeast plus system and growth in CHROMagar candida. The identification of other fungi was based on macromorphologic, microscopic, biochemical and physiological characteristics. Results: The highest frequency of yeasts and mold fungi were observed in Candida albicans 6.6% and Penicillium spp. 25%. The frequency rate of C. neoformans isolation was 2.5%. Conclusion: Several types of fungi are present in pigeon droppings that can spread in environment and transmit to children and elderly as well as immunocompromised patients who are at increased risk of contracting opportunistic diseases.

  7. DNA mutations mediate microevolution between host-adapted forms of the pathogenic fungus Cryptococcus neoformans.

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    Denise A Magditch

    Full Text Available The disease cryptococcosis, caused by the fungus Cryptococcus neoformans, is acquired directly from environmental exposure rather than transmitted person-to-person. One explanation for the pathogenicity of this species is that interactions with environmental predators select for virulence. However, co-incubation of C. neoformans with amoeba can cause a "switch" from the normal yeast morphology to a pseudohyphal form, enabling fungi to survive exposure to amoeba, yet conversely reducing virulence in mammalian models of cryptococcosis. Like other human pathogenic fungi, C. neoformans is capable of microevolutionary changes that influence the biology of the organism and outcome of the host-pathogen interaction. A yeast-pseudohyphal phenotypic switch also happens under in vitro conditions. Here, we demonstrate that this morphological switch, rather than being under epigenetic control, is controlled by DNA mutation since all pseudohyphal strains bear mutations within genes encoding components of the RAM pathway. High rates of isolation of pseudohyphal strains can be explained by the physical size of RAM pathway genes and a hypermutator phenotype of the strain used in phenotypic switching studies. Reversion to wild type yeast morphology in vitro or within a mammalian host can occur through different mechanisms, with one being counter-acting mutations. Infection of mice with RAM mutants reveals several outcomes: clearance of the infection, asymptomatic maintenance of the strains, or reversion to wild type forms and progression of disease. These findings demonstrate a key role of mutation events in microevolution to modulate the ability of a fungal pathogen to cause disease.

  8. Simulated in situ competitive ability and survival of a representative soil yeast, Cryptococcus albidus.

    Science.gov (United States)

    Vishniac, H S

    1995-11-01

    Microcosms containing an air-dried autoclaved loamy sand (Eufala A) with low salt and organic content were inoculated with a representative (obligately aerobic, encapsulated) soil yeast, Cryptococcus albidus var. albidus (T) ATCC 10666, singly (for growth rate and survival determinations) and together with the bacterial biota native to Eufala A. The yeast competed successfully with the more rapidly growing bacteria in the presence of added water from 1% (5.7% of field capacity) to 14% (80% of field capacity) but grew for shorter times than when grown alone; times correlated with the lag phase of the bacterial biota. When well-watered (10 and 14%) competition cultures were allowed to dry and used as inoculum for subcultures, the yeast made significant growth only at 1% added water but survived at the higher moisture concentrations. The competitive ability of Cr. albidus confirms the previously reported advantages of the cryptococcal capsule in hydration and desiccation and, together with lengthy survival, suggests that the importance of such yeasts in the biogeochemistry of arid soils has been seriously underestimated.

  9. Direct transesterification of wet Cryptococcus curvatus cells to biodiesel through use of microwave irradiation

    International Nuclear Information System (INIS)

    Cui, Yi; Liang, Yanna

    2014-01-01

    Highlights: • Direct transesterfication of wet yeast cells using methanol and microwave irradiation is feasible. • Methanol to biomass ratio, stirring speed and KOH concentration were critical to biodiesel yield. • Under optimal conditions, the crude biodiesel contained 64% of FAMEs and was 92% of yeast lipids. - Abstract: Cryptococcus curvatus is a highly promising oleaginous yeast strain that can accumulate intracellular lipids when grown on renewable carbon sources. In order to convert yeast lipids to biodiesel in a simple but cost-effective way, we aim to react whole yeast cells with methanol to produce biodiesel eliminating the step of drying and lipid extraction while adopting microwave energy for heating and disrupting cell walls. Through use of a screening test followed by response surface methodology, optimal parameters leading to the highest yield of crude biodiesel and FAMEs were identified. Under optimal conditions of reaction time (2 min), methanol/biomass ratio (50/1, v/m), stirring speed (966 rpm), KOH concentration (5%), and water content (80%), the yield of crude biodiesel (% of total lipids) was 56.1% after the first round reaction. A second round reaction using the residual yeast cells increased the total yield to 92%. Among the crude biodiesel, 63.88% was FAMEs as revealed by GC analysis. Results from this study indicated that it is feasible to produce biodiesel from wet microbial biomass directly without the steps of drying and lipid extraction. With the assistance of microwave, this process can be accomplished in minutes with good process efficiency

  10. Cryptococcus neoformans capsular enlargement and cellular gigantism during Galleria mellonella infection.

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    Rocío García-Rodas

    Full Text Available We have studied infection of Cryptococcus neoformans in the non-vertebrate host Galleria mellonella with particular interest in the morphological response of the yeast. Inoculation of C. neoformans in caterpillars induced a capsule-independent increase in haemocyte density 2 h after infection. C. neoformans manifested a significant increase in capsule size after inoculation into the caterpillar. The magnitude of capsule increase depended on the temperature, being more pronounced at 37°C than at 30°C, which correlated with an increased virulence of the fungus and reduced phagocytosis at 37°C. Capsule enlargement impaired phagocytosis by haemocytes. Incubation of the yeast in G. mellonella extracts also resulted in capsule enlargement, with the polar lipidic fraction having a prominent role in this effect. During infection, the capsule decreased in permeability. A low proportion of the cells (<5% recovered from caterpillars measured more than 30 µm and were considered giant cells. Giant cells recovered from mice were able to kill the caterpillars in a manner similar to regular cells obtained from in vivo or grown in vitro, establishing their capacity to cause disease. Our results indicate that the morphological transitions exhibited by C. neoformans in mammals also occur in a non-vertebrate host system. The similarities in morphological transitions observed in different animal hosts and in their triggers are consistent with the hypothesis that the cell body and capsular responses represent an adaptation of environmental survival strategies to pathogenesis.

  11. A Case of Cutaneous Plasmablastic Lymphoma in HIV/AIDS with Disseminated Cryptococcus

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    Jun Gong

    2013-01-01

    Full Text Available We present a case of a patient with HIV/AIDS who presented with a tender left lower extremity cutaneous mass over a site of previous cryptococcal infection and was found to have plasmablastic lymphoma (PBL. The incidence of PBL is estimated to account for less than 5% of all cases of non-Hodgkin lymphoma (NHL in HIV-positive individuals. In fact, there were only two reports of extraoral PBL at the time of a 2003 review. PBL in HIV-positive individuals is an aggressive malignancy that tends to occur in middle-aged males with low CD4 counts, high viral loads, and chronic HIV infection. The definitive diagnosis can be made with biopsy which typically shows malignant lymphoid cells that stain positive for plasma cell markers and negative for B-cell markers. The most common treatment is chemotherapy with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP or CHOP-like regimens, but the overall survival rate is poor despite its relative responsiveness to chemotherapy. This case highlights the challenges that remain in improving clinical outcomes, the importance of antiretroviral therapy and HIV disease control, and a potential association between a chronic inflammatory state caused by disseminated Cryptococcus and tumorigenesis in individuals with PBL.

  12. E-NTPDase and E-ADA activities in rats experimental infected by Cryptococcus neoformans.

    Science.gov (United States)

    de Azevedo, Maria Isabel; Ferreiro, Laerte; Da Silva, Aleksandro S; Tonin, Alexandre A; Ruchel, Jader B; Rezer, João F P; França, Raqueli T; Zimmermann, Carine E P; Leal, Daniela B R; Duarte, Marta M M F; Lopes, Sonia T A; Flores, Mariana M; Fighera, Rafael; Santurio, Janio M

    2014-11-07

    Cryptococcus neoformans, the etiological agent of cryptococcosis, is an opportunistic fungal pathogen of immunocompromised individuals. The aim of this study was to evaluate the activities of E-NTPDase and E-ADA in rats experimentally infected by C. neoformans var. grubii. Adult rats (35) were divided in two groups: 18 for the control group (uninfected) (A), and 17 for the infected group (B). Each group was separated into three sub-groups (A1, A2, A3-B1, B2, B3), and samples were collected on 10, 20, and 30 days post-infection (PI). Leukocyte counts, IFN-γ, TNF-α, IgM, IgG levels, and E-NTPDase and E-ADA activities were analyzed. It was possible to observe that IgG and IgM seric levels of infected rats were significantly elevated (PADA activity had a significant reduction (PADA activity in lymphocytes increased significantly (PADA), concomitantly with an inflammatory response (increased levels of cytokines and immunoglobulins) associated with inflammatory infiltrates and histological lesions in the lung. Copyright © 2014 Elsevier B.V. All rights reserved.

  13. Evaluación de varias técnicas de extracción de ADN de Cryptococcus spp. a partir de muestras ambientales.

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    Alexandra Castañeda

    2004-09-01

    Full Text Available El género Cryptococcus comprende, al menos, 38 especies, pero sólo 3 se han informado como patógenas para el hombre y los animales: Cryptococcus laurentii, Cryptococcus albidus y Cryptococcus neoformans; esta última es la más frecuente. La infección se adquiere por la inhalación de los propágulos infectantes del medio ambiente. Los estudios del hábitat se han realizado con técnicas de extracción con soluciones tampón y cultivos en medios selectivos. El objetivo del trabajo fue evaluar varias técnicas de extracción del ADN de Cryptococcus spp. a partir de muestras ambientales. Como controles se emplearon aislamientos de C. neoformans, C. albidus, C. laurentii y Paracoccidiodes brasiliensis. Se emplearon vermiculitas, suelos contaminados en el laboratorio con 10 a 106 blastoconidias/g y muestras naturalmente colonizadas con C. neoformans. El ADN se extrajo con métodos físicos, químicos y con un estuche comercial, y se purificó usando bloques de agarosa y columnas de sílica. Para la amplificación con PCR se emplearon los iniciadores CN4-CN5 específicos para C. neoformans. Sólo el estuche comercial permitió extraer y purificar el ADN de las muestras de suelos contaminados hasta una concentración de 10 blastoconidias/g de suelo y de una de las muestras naturalmente colonizadas. Con este trabajo se logró la extracción y amplificación de ADN de Cryptococcus spp. a partir de muestras ambientales lo cual constituye una herramienta importante para delimitar las áreas ecológicas de C. neoformans en nuestro país.

  14. Conversion of beet molasses and cheese whey into fatty acid methyl esters by the yeast Cryptococcus curvatus.

    Science.gov (United States)

    Takakuwa, Naoya; Saito, Katsuichi

    2010-01-01

    Eighty-one yeast isolates from raw milk were surveyed for the production of fatty acid methyl esters (FAME). Only one species, identified as Cryptococcus curvatus, produced FAME at a detectable level. Cr. curvatus TYC-19 produced more FAME from beet molasses and cheese whey medium than other strains of the same species. In both media, the major FAME produced were linoleic and oleic acid methyl esters. Sequence analysis of the internal transcribed spacer region of ribosomal DNA indicated that TYC-19 diverged from the same species.

  15. Passive immunization against Cryptococcus neoformans with an isotype-switch family of monoclonal antibodies reactive with cryptococcal polysaccharide.

    OpenAIRE

    Sanford, J E; Lupan, D M; Schlageter, A M; Kozel, T R

    1990-01-01

    The in vivo properties of an immunoglobulin isotype-switch family of monoclonal antibodies specific for the polysaccharide capsule of Cryptococcus neoformans were examined in a murine model of cryptococcosis. Subclass-switch variants were isolated by sequential sublining of an immunoglobulin G subclass 1 (IgG1)-secreting cell line. Antibodies of the IgG1, IgG2a, and IgG2b isotypes with identical reactivities with cryptococcal polysaccharide were prepared. The antibodies had the distinct biolo...

  16. ATYPICAL PRESENTATION OF CRYPTOCOCCUS NEOFORMANS IN A KOALA (PHASCOLARCTOS CINEREUS): A MAGNETIC RESONANCE IMAGING AND COMPUTED TOMOGRAPHY STUDY.

    Science.gov (United States)

    Martínez-Nevado, Eva; Alonso-Alegre, Elisa González; Martínez, M Ángeles Jiménez; Rodríguez-Álvaro, Alfonso; de Merlo, Elena Martínez; García, Juncal García; Real, Isabel García

    2017-03-01

    Cryptococcosis is a worldwide and potentially fatal mycosis documented in wild and captive koalas ( Phascolarctos cinereus ) caused by Cryptococcus neoformans . Though mainly a subclinical disease, when the nasal cavity is affected, epistaxis, mucopurulent nasal discharge, dyspnea, and facial distortion may occur. This report describes a case of cryptococcosis in a koala where unilateral exophthalmos was the only evident clinical sign and magnetic resonance imaging and computed tomography findings are described. Both advanced imaging techniques should be considered as standard and complementary techniques for nasal cavity evaluation in koalas.

  17. Interaction of Cryptococcus neoformans Rim101 and protein kinase A regulates capsule.

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    Teresa R O'Meara

    2010-02-01

    Full Text Available Cryptococcus neoformans is a prevalent human fungal pathogen that must survive within various tissues in order to establish a human infection. We have identified the C. neoformans Rim101 transcription factor, a highly conserved pH-response regulator in many fungal species. The rim101 multiply sign in circle mutant strain displays growth defects similar to other fungal species in the presence of alkaline pH, increased salt concentrations, and iron limitation. However, the rim101 multiply sign in circle strain is also characterized by a striking defect in capsule, an important virulence-associated phenotype. This capsular defect is likely due to alterations in polysaccharide attachment to the cell surface, not in polysaccharide biosynthesis. In contrast to many other C. neoformans capsule-defective strains, the rim101 multiply sign in circle mutant is hypervirulent in animal models of cryptococcosis. Whereas Rim101 activation in other fungal species occurs through the conserved Rim pathway, we demonstrate that C. neoformans Rim101 is also activated by the cAMP/PKA pathway. We report here that C. neoformans uses PKA and the Rim pathway to regulate the localization, activation, and processing of the Rim101 transcription factor. We also demonstrate specific host-relevant activating conditions for Rim101 cleavage, showing that C. neoformans has co-opted conserved signaling pathways to respond to the specific niche within the infected host. These results establish a novel mechanism for Rim101 activation and the integration of two conserved signaling cascades in response to host environmental conditions.

  18. Global transcriptome profile of Cryptococcus neoformans during exposure to hydrogen peroxide induced oxidative stress.

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    Rajendra Upadhya

    Full Text Available The ability of the opportunistic fungal pathogen Cryptococcus neoformans to resist oxidative stress is one of its most important virulence related traits. To cope with the deleterious effect of cellular damage caused by the oxidative burst inside the macrophages, C. neoformans has developed multilayered redundant molecular responses to neutralize the stress, to repair the damage and to eventually grow inside the hostile environment of the phagosome. We used microarray analysis of cells treated with hydrogen peroxide (H(2O(2 at multiple time points in a nutrient defined medium to identify a transcriptional signature associated with oxidative stress. We discovered that the composition of the medium in which fungal cells were grown and treated had a profound effect on their capacity to degrade exogenous H(2O(2. We determined the kinetics of H(2O(2 breakdown by growing yeast cells under different conditions and accordingly selected an appropriate media composition and range of time points for isolating RNA for hybridization. Microarray analysis revealed a robust transient transcriptional response and the intensity of the global response was consistent with the kinetics of H(2O(2 breakdown by treated cells. Gene ontology analysis of differentially expressed genes related to oxidation-reduction, metabolic process and protein catabolic processes identified potential roles of mitochondrial function and protein ubiquitination in oxidative stress resistance. Interestingly, the metabolic pathway adaptation of C. neoformans to H(2O(2 treatment was remarkably distinct from the response of other fungal organisms to oxidative stress. We also identified the induction of an antifungal drug resistance response upon the treatment of C. neoformans with H(2O(2. These results highlight the complexity of the oxidative stress response and offer possible new avenues for improving our understanding of mechanisms of oxidative stress resistance in C. neoformans.

  19. Unisexual reproduction drives meiotic recombination and phenotypic and karyotypic plasticity in Cryptococcus neoformans.

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    Sheng Sun

    2014-12-01

    Full Text Available In fungi, unisexual reproduction, where sexual development is initiated without the presence of two compatible mating type alleles, has been observed in several species that can also undergo traditional bisexual reproduction, including the important human fungal pathogens Cryptococcus neoformans and Candida albicans. While unisexual reproduction has been well characterized qualitatively, detailed quantifications are still lacking for aspects of this process, such as the frequency of recombination during unisexual reproduction, and how this compares with bisexual reproduction. Here, we analyzed meiotic recombination during α-α unisexual and a-α bisexual reproduction of C. neoformans. We found that meiotic recombination operates in a similar fashion during both modes of sexual reproduction. Specifically, we observed that in α-α unisexual reproduction, the numbers of crossovers along the chromosomes during meiosis, recombination frequencies at specific chromosomal regions, as well as meiotic recombination hot and cold spots, are all similar to those observed during a-α bisexual reproduction. The similarity in meiosis is also reflected by the fact that phenotypic segregation among progeny collected from the two modes of sexual reproduction is also similar, with transgressive segregation being observed in both. Additionally, we found diploid meiotic progeny were also produced at similar frequencies in the two modes of sexual reproduction, and transient chromosomal loss and duplication likely occurs frequently and results in aneuploidy and loss of heterozygosity that can span entire chromosomes. Furthermore, in both α-α unisexual and a-α bisexual reproduction, we observed biased allele inheritance in regions on chromosome 4, suggesting the presence of fragile chromosomal regions that might be vulnerable to mitotic recombination. Interestingly, we also observed a crossover event that occurred within the MAT locus during α-α unisexual

  20. Factors Required for Activation of Urease as a Virulence Determinant in Cryptococcus neoformans

    Science.gov (United States)

    Singh, Arpita; Panting, Robert J.; Varma, Ashok; Saijo, Tomomi; Waldron, Kevin J.; Jong, Ambrose; Ngamskulrungroj, Popchai; Chang, Yun C.; Rutherford, Julian C.; Kwon-Chung, Kyung J.

    2013-01-01

    ABSTRACT Urease in Cryptococcus neoformans plays an important role in fungal dissemination to the brain and causing meningoencephalitis. Although urea is not required for synthesis of apourease encoded by URE1, the available nitrogen source affected the expression of URE1 as well as the level of the enzyme activity. Activation of the apoenzyme requires three accessory proteins, Ure4, Ure6, and Ure7, which are homologs of the bacterial urease accessory proteins UreD, UreF, and UreG, respectively. A yeast two-hybrid assay showed positive interaction of Ure1 with the three accessory proteins encoded by URE4, URE6, and URE7. Metalloproteomic analysis of cryptococcal lysates using inductively coupled plasma mass spectrometry (ICP-MS) and a biochemical assay of urease activity showed that, as in many other organisms, urease is a metallocentric enzyme that requires nickel transported by Nic1 for its catalytic activity. The Ure7 accessory protein (bacterial UreG homolog) binds nickel likely via its conserved histidine-rich domain and appears to be responsible for the incorporation of Ni2+ into the apourease. Although the cryptococcal genome lacks the bacterial UreE homolog, Ure7 appears to combine the functions of bacterial UreE and UreG, thus making this pathogen more similar to that seen with the plant system. Brain invasion by the ure1, ure7, and nic1 mutant strains that lack urease activity was significantly less effective in a mouse model. This indicated that an activated urease and not the Ure1 protein was responsible for enhancement of brain invasion and that the factors required for urease activation in C. neoformans resemble those of plants more than those of bacteria. PMID:23653445

  1. Surface-associated plasminogen binding of Cryptococcus neoformans promotes extracellular matrix invasion.

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    Jamal Stie

    2009-06-01

    Full Text Available The fungal pathogen Cryptococcus neoformans is a leading cause of illness and death in persons with predisposing factors, including: malignancies, solid organ transplants, and corticosteroid use. C. neoformans is ubiquitous in the environment and enters into the lungs via inhalation, where it can disseminate through the bloodstream and penetrate the central nervous system (CNS, resulting in a difficult to treat and often-fatal infection of the brain, called meningoencephalitis. Plasminogen is a highly abundant protein found in the plasma component of blood and is necessary for the degradation of fibrin, collagen, and other structural components of tissues. This fibrinolytic system is utilized by cancer cells during metastasis and several pathogenic species of bacteria have been found to manipulate the host plasminogen system to facilitate invasion of tissues during infection by modifying the activation of this process through the binding of plasminogen at their surface.The invasion of the brain and the central nervous system by penetration of the protective blood-brain barrier is a prerequisite to the establishment of meningoencephalitis by the opportunistic fungal pathogen C. neoformans. In this study, we examined the ability of C. neoformans to subvert the host plasminogen system to facilitate tissue barrier invasion. Through a combination of biochemical, cell biology, and proteomic approaches, we have shown that C. neoformans utilizes the host plasminogen system to cross tissue barriers, providing support for the hypothesis that plasminogen-binding may contribute to the invasion of the blood-brain barrier by penetration of the brain endothelial cells and underlying matrix. In addition, we have identified the cell wall-associated proteins that serve as plasminogen receptors and characterized both the plasminogen-binding and plasmin-activation potential for this significant human pathogen.The results of this study provide evidence for the

  2. Nitrogen Metabolite Repression of Metabolism and Virulence in the Human Fungal Pathogen Cryptococcus neoformans

    Science.gov (United States)

    Lee, I. Russel; Chow, Eve W. L.; Morrow, Carl A.; Djordjevic, Julianne T.; Fraser, James A.

    2011-01-01

    Proper regulation of metabolism is essential to maximizing fitness of organisms in their chosen environmental niche. Nitrogen metabolite repression is an example of a regulatory mechanism in fungi that enables preferential utilization of easily assimilated nitrogen sources, such as ammonium, to conserve resources. Here we provide genetic, transcriptional, and phenotypic evidence of nitrogen metabolite repression in the human pathogen Cryptococcus neoformans. In addition to loss of transcriptional activation of catabolic enzyme-encoding genes of the uric acid and proline assimilation pathways in the presence of ammonium, nitrogen metabolite repression also regulates the production of the virulence determinants capsule and melanin. Since GATA transcription factors are known to play a key role in nitrogen metabolite repression, bioinformatic analyses of the C. neoformans genome were undertaken and seven predicted GATA-type genes were identified. A screen of these deletion mutants revealed GAT1, encoding the only global transcription factor essential for utilization of a wide range of nitrogen sources, including uric acid, urea, and creatinine—three predominant nitrogen constituents found in the C. neoformans ecological niche. In addition to its evolutionarily conserved role in mediating nitrogen metabolite repression and controlling the expression of catabolic enzyme and permease-encoding genes, Gat1 also negatively regulates virulence traits, including infectious basidiospore production, melanin formation, and growth at high body temperature (39°–40°). Conversely, Gat1 positively regulates capsule production. A murine inhalation model of cryptococcosis revealed that the gat1Δ mutant is slightly more virulent than wild type, indicating that Gat1 plays a complex regulatory role during infection. PMID:21441208

  3. Allelic exchange of pheromones and their receptors reprograms sexual identity in Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Brynne C Stanton

    2010-02-01

    Full Text Available Cell type specification is a fundamental process that all cells must carry out to ensure appropriate behaviors in response to environmental stimuli. In fungi, cell identity is critical for defining "sexes" known as mating types and is controlled by components of mating type (MAT loci. MAT-encoded genes function to define sexes via two distinct paradigms: 1 by controlling transcription of components common to both sexes, or 2 by expressing specially encoded factors (pheromones and their receptors that differ between mating types. The human fungal pathogen Cryptococcus neoformans has two mating types (a and alpha that are specified by an extremely unusual MAT locus. The complex architecture of this locus makes it impossible to predict which paradigm governs mating type. To identify the mechanism by which the C. neoformans sexes are determined, we created strains in which the pheromone and pheromone receptor from one mating type (a replaced the pheromone and pheromone receptor of the other (alpha. We discovered that these "alpha(a" cells effectively adopt a new mating type (that of a cells; they sense and respond to alpha factor, they elicit a mating response from alpha cells, and they fuse with alpha cells. In addition, alpha(a cells lose the alpha cell type-specific response to pheromone and do not form germ tubes, instead remaining spherical like a cells. Finally, we discovered that exogenous expression of the diploid/dikaryon-specific transcription factor Sxi2a could then promote complete sexual development in crosses between alpha and alpha(a strains. These data reveal that cell identity in C. neoformans is controlled fully by three kinds of MAT-encoded proteins: pheromones, pheromone receptors, and homeodomain proteins. Our findings establish the mechanisms for maintenance of distinct cell types and subsequent developmental behaviors in this unusual human fungal pathogen.

  4. Characterization of the complete uric acid degradation pathway in the fungal pathogen Cryptococcus neoformans.

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    I Russel Lee

    Full Text Available Degradation of purines to uric acid is generally conserved among organisms, however, the end product of uric acid degradation varies from species to species depending on the presence of active catabolic enzymes. In humans, most higher primates and birds, the urate oxidase gene is non-functional and hence uric acid is not further broken down. Uric acid in human blood plasma serves as an antioxidant and an immune enhancer; conversely, excessive amounts cause the common affliction gout. In contrast, uric acid is completely degraded to ammonia in most fungi. Currently, relatively little is known about uric acid catabolism in the fungal pathogen Cryptococcus neoformans even though this yeast is commonly isolated from uric acid-rich pigeon guano. In addition, uric acid utilization enhances the production of the cryptococcal virulence factors capsule and urease, and may potentially modulate the host immune response during infection. Based on these important observations, we employed both Agrobacterium-mediated insertional mutagenesis and bioinformatics to predict all the uric acid catabolic enzyme-encoding genes in the H99 genome. The candidate C. neoformans uric acid catabolic genes identified were named: URO1 (urate oxidase, URO2 (HIU hydrolase, URO3 (OHCU decarboxylase, DAL1 (allantoinase, DAL2,3,3 (allantoicase-ureidoglycolate hydrolase fusion protein, and URE1 (urease. All six ORFs were then deleted via homologous recombination; assaying of the deletion mutants' ability to assimilate uric acid and its pathway intermediates as the sole nitrogen source validated their enzymatic functions. While Uro1, Uro2, Uro3, Dal1 and Dal2,3,3 were demonstrated to be dispensable for virulence, the significance of using a modified animal model system of cryptococcosis for improved mimicking of human pathogenicity is discussed.

  5. Unisexual Reproduction Drives Meiotic Recombination and Phenotypic and Karyotypic Plasticity in Cryptococcus neoformans

    Science.gov (United States)

    Sun, Sheng; Billmyre, R. Blake; Mieczkowski, Piotr A.; Heitman, Joseph

    2014-01-01

    In fungi, unisexual reproduction, where sexual development is initiated without the presence of two compatible mating type alleles, has been observed in several species that can also undergo traditional bisexual reproduction, including the important human fungal pathogens Cryptococcus neoformans and Candida albicans. While unisexual reproduction has been well characterized qualitatively, detailed quantifications are still lacking for aspects of this process, such as the frequency of recombination during unisexual reproduction, and how this compares with bisexual reproduction. Here, we analyzed meiotic recombination during α-α unisexual and a-α bisexual reproduction of C. neoformans. We found that meiotic recombination operates in a similar fashion during both modes of sexual reproduction. Specifically, we observed that in α-α unisexual reproduction, the numbers of crossovers along the chromosomes during meiosis, recombination frequencies at specific chromosomal regions, as well as meiotic recombination hot and cold spots, are all similar to those observed during a-α bisexual reproduction. The similarity in meiosis is also reflected by the fact that phenotypic segregation among progeny collected from the two modes of sexual reproduction is also similar, with transgressive segregation being observed in both. Additionally, we found diploid meiotic progeny were also produced at similar frequencies in the two modes of sexual reproduction, and transient chromosomal loss and duplication likely occurs frequently and results in aneuploidy and loss of heterozygosity that can span entire chromosomes. Furthermore, in both α-α unisexual and a-α bisexual reproduction, we observed biased allele inheritance in regions on chromosome 4, suggesting the presence of fragile chromosomal regions that might be vulnerable to mitotic recombination. Interestingly, we also observed a crossover event that occurred within the MAT locus during α-α unisexual reproduction. Our results

  6. Protein Composition of Infectious Spores Reveals Novel Sexual Development and Germination Factors in Cryptococcus.

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    Mingwei Huang

    2015-08-01

    Full Text Available Spores are an essential cell type required for long-term survival across diverse organisms in the tree of life and are a hallmark of fungal reproduction, persistence, and dispersal. Among human fungal pathogens, spores are presumed infectious particles, but relatively little is known about this robust cell type. Here we used the meningitis-causing fungus Cryptococcus neoformans to determine the roles of spore-resident proteins in spore biology. Using highly sensitive nanoscale liquid chromatography/mass spectrometry, we compared the proteomes of spores and vegetative cells (yeast and identified eighteen proteins specifically enriched in spores. The genes encoding these proteins were deleted, and the resulting strains were evaluated for discernable phenotypes. We hypothesized that spore-enriched proteins would be preferentially involved in spore-specific processes such as dormancy, stress resistance, and germination. Surprisingly, however, the majority of the mutants harbored defects in sexual development, the process by which spores are formed. One mutant in the cohort was defective in the spore-specific process of germination, showing a delay specifically in the initiation of vegetative growth. Thus, by using this in-depth proteomics approach as a screening tool for cell type-specific proteins and combining it with molecular genetics, we successfully identified the first germination factor in C. neoformans. We also identified numerous proteins with previously unknown functions in both sexual development and spore composition. Our findings provide the first insights into the basic protein components of infectious spores and reveal unexpected molecular connections between infectious particle production and spore composition in a pathogenic eukaryote.

  7. Allelic exchange of pheromones and their receptors reprograms sexual identity in Cryptococcus neoformans.

    Science.gov (United States)

    Stanton, Brynne C; Giles, Steven S; Staudt, Mark W; Kruzel, Emilia K; Hull, Christina M

    2010-02-26

    Cell type specification is a fundamental process that all cells must carry out to ensure appropriate behaviors in response to environmental stimuli. In fungi, cell identity is critical for defining "sexes" known as mating types and is controlled by components of mating type (MAT) loci. MAT-encoded genes function to define sexes via two distinct paradigms: 1) by controlling transcription of components common to both sexes, or 2) by expressing specially encoded factors (pheromones and their receptors) that differ between mating types. The human fungal pathogen Cryptococcus neoformans has two mating types (a and alpha) that are specified by an extremely unusual MAT locus. The complex architecture of this locus makes it impossible to predict which paradigm governs mating type. To identify the mechanism by which the C. neoformans sexes are determined, we created strains in which the pheromone and pheromone receptor from one mating type (a) replaced the pheromone and pheromone receptor of the other (alpha). We discovered that these "alpha(a)" cells effectively adopt a new mating type (that of a cells); they sense and respond to alpha factor, they elicit a mating response from alpha cells, and they fuse with alpha cells. In addition, alpha(a) cells lose the alpha cell type-specific response to pheromone and do not form germ tubes, instead remaining spherical like a cells. Finally, we discovered that exogenous expression of the diploid/dikaryon-specific transcription factor Sxi2a could then promote complete sexual development in crosses between alpha and alpha(a) strains. These data reveal that cell identity in C. neoformans is controlled fully by three kinds of MAT-encoded proteins: pheromones, pheromone receptors, and homeodomain proteins. Our findings establish the mechanisms for maintenance of distinct cell types and subsequent developmental behaviors in this unusual human fungal pathogen.

  8. Surface localization of glucosylceramide during Cryptococcus neoformans infection allows targeting as a potential antifungal.

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    Ryan Rhome

    Full Text Available Cryptococcus neoformans (Cn is a significant human pathogen that, despite current treatments, continues to have a high morbidity rate especially in sub-Saharan Africa. The need for more tolerable and specific therapies has been clearly shown. In the search for novel drug targets, the gene for glucosylceramide synthase (GCS1 was deleted in Cn, resulting in a strain (Δgcs1 that does not produce glucosylceramide (GlcCer and is avirulent in mouse models of infection. To understand the biology behind the connection between virulence and GlcCer, the production and localization of GlcCer must be characterized in conditions that are prohibitive to the growth of Δgcs1 (neutral pH and high CO(2. These prohibitive conditions are physiologically similar to those found in the extracellular spaces of the lung during infection. Here, using immunofluorescence, we have shown that GlcCer localization to the cell surface is significantly increased during growth in these conditions and during infection. We further seek to exploit this localization by treatment with Cerezyme (Cz, a recombinant enzyme that metabolizes GlcCer, as a potential treatment for Cn. Cz treatment was found to reduce the amount of GlcCer in vitro, in cultures, and in Cn cells inhabiting the mouse lung. Treatment with Cz induced a membrane integrity defect in wild type Cn cells similar to Δgcs1. Cz treatment also reduced the in vitro growth of Cn in a dose and condition dependent manner. Finally, Cz treatment was shown to have a protective effect on survival in mice infected with Cn. Taken together, these studies have established the legitimacy of targeting the GlcCer and other related sphingolipid systems in the development of novel therapeutics.

  9. Rapid direct identification of Cryptococcus neoformans from pigeon droppings by nested PCR using CNLAC1 gene.

    Science.gov (United States)

    Chae, H S; Park, G N; Kim, S H; Jo, H J; Kim, J T; Jeoung, H Y; An, D J; Kim, N H; Shin, B W; Kang, Y I; Chang, K S

    2012-08-01

    Isolation and identification of Cryptococcus neoformans and pathogenic yeast-like fungi from pigeon droppings has been taken for a long time and requires various nutrients for its growth. In this study, we attempted to establish a rapid direct identification method of Cr. neoformans from pigeon dropping samples by nested-PCR using internal transcribed spacer (ITS) CAP64 and CNLAC1 genes, polysaccharide capsule gene and laccase-associated gene to produce melanin pigment, respectively, which are common genes of yeasts. The ITS and CAP64 genes were amplified in all pathogenic yeasts, but CNLAC1 was amplified only in Cr. neoformans. The ITS gene was useful for yeast genotyping depending on nucleotide sequence. Homology of CAP64 genes among the yeasts were very high. The specificity of PCR using CNLAC1 was demonstrated in Cr. neoformans environmental strains but not in other yeast-like fungi. The CNLAC1 gene was detected in 5 serotypes of Cr. neoformans. The nested-PCR amplified up to 10(-11) μg of the genomic DNA and showed high sensitivity. All pigeon droppings among 31 Cr. neoformans-positive samples were positive and all pigeon droppings among 348 Cr. neoformans-negative samples were negative by the direct nested-PCR. In addition, after primary enrichment of pigeon droppings in Sabouraud dextrose broth, all Cr. neoformans-negative samples were negative by the nested-PCR, which showed high specificity. The nested-PCR showed high sensitivity without culture of pigeon droppings. Nested-PCR using CNLAC1 provides a rapid and reliable molecular diagnostic method to overcome weak points such as long culture time of many conventional methods.

  10. Effects of radiation type and delivery mode on a radioresistant eukaryote Cryptococcus neoformans

    International Nuclear Information System (INIS)

    Shuryak, Igor; Bryan, Ruth A.; Broitman, Jack; Marino, Stephen A.; Morgenstern, Alfred; Apostolidis, Christos; Dadachova, Ekaterina

    2015-01-01

    Introduction: Most research on radioresistant fungi, particularly on human pathogens such as Cryptococcus neoformans, involves sparsely-ionizing radiation. Consequently, fungal responses to densely-ionizing radiation, which can be harnessed to treat life-threatening fungal infections, remain incompletely understood. Methods: We addressed this issue by quantifying and comparing the effects of densely-ionizing α-particles (delivered either by external beam or by 213 Bi-labeled monoclonal antibodies), and sparsely-ionizing 137 Cs γ-rays, on Cryptococus neoformans. Results: The best-fit linear-quadratic parameters for clonogenic survival were the following: α = 0.24 × 10 −2 Gy −1 for γ-rays and 1.07 × 10 −2 Gy −1 for external-beam α-particles, and β = 1.44 × 10 −5 Gy −2 for both radiation types. Fungal cell killing by radiolabeled antibodies was consistent with predictions based on the α-particle dose to the cell nucleus and the linear-quadratic parameters for external-beam α-particles. The estimated RBE (for α-particles vs. γ-rays) at low doses was 4.47 for the initial portion of the α-particle track, and 7.66 for the Bragg peak. Non-radiological antibody effects accounted for up to 23% of cell death. Conclusions: These results quantify the degree of C. neoformans resistance to densely-ionizing radiations, and show how this resistance can be overcome with fungus-specific radiolabeled antibodies

  11. HapX positively and negatively regulates the transcriptional response to iron deprivation in Cryptococcus neoformans.

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    Won Hee Jung

    2010-11-01

    Full Text Available The fungal pathogen Cryptococcus neoformans is a major cause of illness in immunocompromised individuals such as AIDS patients. The ability of the fungus to acquire nutrients during proliferation in host tissue and the ability to elaborate a polysaccharide capsule are critical determinants of disease outcome. We previously showed that the GATA factor, Cir1, is a major regulator both of the iron uptake functions needed for growth in host tissue and the key virulence factors such as capsule, melanin and growth at 37°C. We are interested in further defining the mechanisms of iron acquisition from inorganic and host-derived iron sources with the goal of understanding the nutritional adaptation of C. neoformans to the host environment. In this study, we investigated the roles of the HAP3 and HAPX genes in iron utilization and virulence. As in other fungi, the C. neoformans Hap proteins negatively influence the expression of genes encoding respiratory and TCA cycle functions under low-iron conditions. However, we also found that HapX plays both positive and negative roles in the regulation of gene expression, including a positive regulatory role in siderophore transporter expression. In addition, HapX also positively regulated the expression of the CIR1 transcript. This situation is in contrast to the negative regulation by HapX of genes encoding GATA iron regulatory factors in Aspergillus nidulans and Schizosaccharomyces pombe. Although both hapX and hap3 mutants were defective in heme utilization in culture, only HapX made a contribution to virulence, and loss of HapX in a strain lacking the high-affinity iron uptake system did not cause further attenuation of disease. Therefore, HapX appears to have a minimal role during infection of mammalian hosts and instead may be an important regulator of environmental iron uptake functions. Overall, these results indicated that C. neoformans employs multiple strategies for iron acquisition during infection.

  12. alpha AD alpha hybrids of Cryptococcus neoformans: evidence of same-sex mating in nature and hybrid fitness.

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    Xiaorong Lin

    2007-10-01

    Full Text Available Cryptococcus neoformans is a ubiquitous human fungal pathogen that causes meningoencephalitis in predominantly immunocompromised hosts. The fungus is typically haploid, and sexual reproduction involves two individuals with opposite mating types/sexes, alpha and a. However, the overwhelming predominance of mating type (MAT alpha over a in C. neoformans populations limits alpha-a mating in nature. Recently it was discovered that C. neoformans can undergo same-sex mating under laboratory conditions, especially between alpha isolates. Whether same-sex mating occurs in nature and contributes to the current population structure was unknown. In this study, natural alpha AD alpha hybrids that arose by fusion between two alpha cells of different serotypes (A and D were identified and characterized, providing definitive evidence that same-sex mating occurs naturally. A novel truncated allele of the mating-type-specific cell identity determinant SXI1 alpha was also identified as a genetic factor likely involved in this process. In addition, laboratory-constructed alpha AD alpha strains exhibited hybrid vigor both in vitro and in vivo, providing a plausible explanation for their relative abundance in nature despite the fact that AD hybrids are inefficient in meiosis/sporulation and are trapped in the diploid state. These findings provide insights on the origins, genetic mechanisms, and fitness impact of unisexual hybridization in the Cryptococcus population.

  13. Ultrastructural and chemotaxonomic analysis of a xylanolytic strain of Cryptococcus adeliensis isolated from sheep droppings in Spain.

    Science.gov (United States)

    Velázquez, Encarna; del Villar, María; Grondona, Isabel; Monte, Enrique; González-Villa, Tomás

    2006-09-01

    Cryptococcus adeliensis was initially described as a psycrophilic species containing a single strain CBS 8351(T) isolated from decayed algae in Terre Adelie (Antartida). Later, a second strain of this species was isolated from an immunosuppressed patient affected by leukaemia in Germany and recently several strains from this species have been found in human patients and pigeon droppings of the same country. In this study, we isolated from sheep droppings in Spain a xylanolytic strain named LEVX01 that was phenotypically related to the strain CBS 8351(T) and showed a 100% similarity in the D1/D2 domain and 5.8S-ITS region sequences with respect to the remaining described strains of C. adeliensis. These findings suggest that this species has a wide geographical distribution and that the animal faeces are a common habitat for C. adeliensis. The chemotaxonomic analyses showed the absence of detectable amounts of xylose in the cell walls of the strains LEVX01 and CBS8351(T) in contrast to other Cryptococcus species. Interestingly, the ultrastructural study showed the presence of fimbriae in these two strains that could be involved in the attachment to the host cells and, as occurs in Candida albicans, they could also be a pathogenicity factor for the man.

  14. MLST-Based Population Genetic Analysis in a Global Context Reveals Clonality amongst Cryptococcus neoformans var. grubii VNI Isolates from HIV Patients in Southeastern Brazil

    NARCIS (Netherlands)

    Ferreira-Paim, Kennio; Andrade-Silva, Leonardo; Fonseca, Fernanda; Ferreira, Thatiana; Mora, Delio; Andrade-Silva, Juliana; Khan, Aziza; Dao, Aiken; C. Reis, Eduardo; Almeida, Margarete; Maltos, André; Rodrigues Jr, Virmondes; Trilles, Luciana; Rickerts, Volker; Chindamporn, Ariya; Sykes, Jane; Cogliati, Massimo; Nielsen, Kirsten; Boekhout, Teun; L. Silva-Vergara, Mario

    2017-01-01

    Cryptococcosis is an important fungal infection in immunocompromised individuals, especially those infected with HIV. In Brazil, despite the free availability of antiretroviral therapy (ART) in the public health system, the mortality rate due to Cryptococcus neoformans meningitis is still high. To

  15. Classification of Cryptococcus neoformans and yeast-like fungus isolates from pigeon droppings by colony phenotyping and ITS genotyping and their seasonal variations in Korea.

    Science.gov (United States)

    Chae, H S; Jang, G E; Kim, N H; Son, H R; Lee, J H; Kim, S H; Park, G N; Jo, H J; Kim, J T; Chang, K S

    2012-03-01

    Cryptococcus neoformans (C neoformans) is a frequent cause of invasive fungal disease in immunocompromised human hosts. Ninety-eight samples of pigeon droppings were collected from the pigeon shelters in Seoul, and cultured on birdseed agar (BSA) and Sabouraud dextrose agar (SDA). One hundred yeast-like colonies were selected and identified via phenotype characteristics, such as colony morphology and biochemical characteristics. This was then followed with genotyping via sequencing of the internal transcribed spacer (ITS) region. The colonies were classified into four kinds of colony color types: brown type (BrT), beige type (BeT), pink type (PT), and white type (WT). Numbers of isolated BrT, BeT, PT, and WT colonies were 22 (22%), 30 (30%), 19 (19%), and 39 (39%), respectively. All BrT colonies were identified as C neoformans. BeT were identified as 19 isolates of Cryptococcus laurentii, 10 isolates of Malassezia furfur, and 1 isolate of Cryptococcus uniguttulatus. PT was divided into two colony color types: light-PT (l-PT) and deep-PT (d-PT). Eighteen of l-PT and one of d-PT were identified as Rhodotorula glutinis and Rhodotorula mucilaginosa, respectively. WT were identified as 34 isolates of Cryptococcus guilliermondii, 3 isolates of Cryptococcus zeylanoides, 1 isolate of Cryptococcus sake, and 1 isolate of Stephanoascus ciferrii. Most strains were classified identically with the use of either phenotype or genotyping techniques, but C uniguttulatus and C sake classified by phenotyping were Pseudozyma aphidis and Cryptococcus famata by genotyping. This rapid screening technique of pathogenic yeast-like fungi by only colony characteristics is also expected to be very useful for primary yeast screening. Additionally, we investigated the seasonal variations of C neoformans and other yeast-like fungi from 379 pigeon-dropping samples that were collected from February 2011 to March 2011. We isolated 685 yeast-like fungi from the samples. Almost all C neoformans and

  16. Two Distinct Approaches for CRISPR-Cas9-Mediated Gene Editing in Cryptococcus neoformans and Related Species.

    Science.gov (United States)

    Wang, Ping

    2018-06-27

    Cryptococcus neoformans and related species are encapsulated basidiomycetous fungi that cause meningoencephalitis in individuals with immune deficiency. This pathogen has a tractable genetic system; however, gene disruption via electroporation remains difficult, while biolistic transformation is often limited by lack of multiple genetic markers and the high initial cost of equipment. The approach using clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) has become the technology of choice for gene editing in many organisms due to its simplicity, efficiency, and versatility. The technique has been successfully demonstrated in C. neoformans and Cryptococcus deneoformans in which two DNA plasmids expressing either the Streptococcus pyogenes CAS9 gene or the guide RNA (gRNA) were employed. However, potential adverse effects due to constitutive expression and the time-consuming process of constructing vectors to express each gRNA remain as a primary barrier for wide adaptation. This report describes the delivery of preassembled CRISPR-Cas9-gRNA ribonucleoproteins (RNPs) via electroporation that is able to generate edited mutant alleles. RNP-mediated CRISPR-Cas9 was used to replace the wild-type GIB2 gene encoding a Gβ-like/RACK1 Gib2 protein with a gib2 :: NAT allele via homologous recombination in both C. neoformans and C. deneoformans In addition, a DNA plasmid (pCnCas9:U6-gRNA) that expresses both Cas9 and gRNA, allowing for convenient yet low-cost DNA-mediated gene editing, is described. pCnCas9:U6-gRNA contains an endogenous U6 promoter for gRNA expression and restriction sites for one-step insertion of a gRNA. These approaches and resources provide new opportunities to accelerate genetic studies of Cryptococcus species. IMPORTANCE For genetic studies of the Cryptococcus genus, generation of mutant strains is often hampered by a limited number of selectable genetic markers, the tedious process of vector

  17. Transmembrane transporter expression regulated by the glucosylceramide pathway in Cryptococcus neoformans.

    Science.gov (United States)

    Singh, Arpita; Rella, Antonella; Schwacke, John; Vacchi-Suzzi, Caterina; Luberto, Chiara; Del Poeta, Maurizio

    2015-11-16

    disrupt transmembrane signaling complex, which in turn contributes to cryptococcal osmotic, pH, ion homeostasis and its pathobiology. Six genes identified from gene expression microarrays by gene set enrichment analysis and validated by RT-PCR, are membrane located and associated with the growth defect at neutral-alkaline pH due to the absence and or presence of a structurally modified GlcCer. They may be involved in the transmembrane signaling network in Cryptococcus neoformans, and therefore the pathobiology of the fungus in these conditions.

  18. N-acetylglucosamine affects Cryptococcus neoformans cell-wall composition and melanin architecture.

    Science.gov (United States)

    Camacho, Emma; Chrissian, Christine; Cordero, Radames J B; Liporagi-Lopes, Livia; Stark, Ruth E; Casadevall, Arturo

    2017-11-01

    Cryptococcus neoformans is an environmental fungus that belongs to the phylum Basidiomycetes and is a major pathogen in immunocompromised patients. The ability of C. neoformans to produce melanin pigments represents its second most important virulence factor, after the presence of a polysaccharide capsule. Both the capsule and melanin are closely associated with the fungal cell wall, a complex structure that is essential for maintaining cell morphology and viability under conditions of stress. The amino sugar N-acetylglucosamine (GlcNAc) is a key constituent of the cell-wall chitin and is used for both N-linked glycosylation and GPI anchor synthesis. Recent studies have suggested additional roles for GlcNAc as an activator and mediator of cellular signalling in fungal and plant cells. Furthermore, chitin and chitosan polysaccharides interact with melanin pigments in the cell wall and have been found to be essential for melanization. Despite the importance of melanin, its molecular structure remains unresolved; however, we previously obtained critical insights using advanced nuclear magnetic resonance (NMR) and imaging techniques. In this study, we investigated the effect of GlcNAc supplementation on cryptococcal cell-wall composition and melanization. C. neoformans was able to metabolize GlcNAc as a sole source of carbon and nitrogen, indicating a capacity to use a component of a highly abundant polymer in the biospherenutritionally. C. neoformans cells grown with GlcNAc manifested changes in the chitosan cell-wall content, cell-wall thickness and capsule size. Supplementing cultures with isotopically 15 N-labelled GlcNAc demonstrated that the exogenous monomer serves as a building block for chitin/chitosan and is incorporated into the cell wall. The altered chitin-to-chitosan ratio had no negative effects on the mother-daughter cell separation; growth with GlcNAc affected the fungal cell-wall scaffold, resulting in increased melanin deposition and assembly. In

  19. The Crucial Role of Biofilms in Cryptococcus neoformans Survival within Macrophages and Colonization of the Central Nervous System

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    Lilit Aslanyan

    2017-02-01

    Full Text Available Cryptococcus neoformans is an encapsulated yeast-like fungus capable of causing life threatening meningoencephalitis in patients with impaired immunity. This microbe primarily infects the host via inhalation but has the ability to disseminate to the central nervous system (CNS either as a single cell or inside of macrophages. Upon traversing the blood brain barrier, C. neoformans has the capacity to form biofilm-like structures known as cryptococcomas. Hence, we will discuss the C. neoformans elements contributing to biofilm formation including the fungus’ ability to survive in the acidic environment of a macrophage phagosome and inside of the CNS. The purpose of this mini-review is to instill fresh interest in understanding the importance of biofilms on fungal pathogenesis.

  20. CRYPTOCOCCUS NEOFORMANS VAR. GRUBII-ASSOCIATED RENAL AMYLOIDOSIS CAUSING PROTEIN-LOSING NEPHROPATHY IN A RED KANGAROO (MACROPUS RUFUS).

    Science.gov (United States)

    Thurber, Mary Irene; Gjeltema, Jenessa; Sheley, Matthew; Wack, Ray F

    2017-09-01

    A 10-year-old male castrated red kangaroo (Macropus rufus) presented with mandibular swelling. Examination findings included pitting edema with no dental disease evident on examination or radiographs. The results of blood work were moderate azotemia, hypoalbuminemia, and severely elevated urine protein:creatinine ratio (9.9). Radiographs showed an interstitial pattern of the caudal right lung, and an abdominal ultrasound demonstrated scant effusion. Symptomatic and empirical therapy with antibiotics, anti-inflammatory drugs, and an angiotensin-converting enzyme (ACE) inhibitor did not resolve clinical signs. Due to poor prognosis and declining quality of life, euthanasia was elected. Necropsy revealed chronic granulomatous pneumonia of the caudal right lung lobe with intralesional Cryptococcus, identified as C. neoformans var. grubii by DNA sequencing. Severe bilateral glomerular and tubulointerstitial amyloidosis induced protein-losing nephropathy, leading to tri-cavitary effusion, subcutaneous edema, and cachexia. The authors speculate that renal amyloidosis was associated with chronic cryptococcal pneumonia in this red kangaroo.

  1. Isolation of human fungi from soil and identification of two endemic areas of Cryptococcus neoformans and Coccidioides immitis

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    Héctor Rubinstein

    1989-02-01

    Full Text Available The present study was carried out in two different areas of Province of Cordoba, Argentina, where there was a suspicious of endemic mycosis. The previous data were the presence of a clinical case of pulmonary cryptococcosis in one area (Alta Gracia and the previous findings of a high incidence of coccidioidin and cryptococcin reactors in the population of the second one (Villa Dolores. In both areas soil samples for fungi were studied and Cryptococcus neoformans was found in 2/25 samples from Alta Gracia. In Villa Dolores Coccidioides immitis was isolated in 2/40 samples, and C. neoformans in 1/40 samples. Delayed hypersensitivity test with cryptococcin was determined in the population from Alta Gracia and it was found to be 5.3%. Positive cutaneous tests with coccidioidin (33.8% and cryptococcin (31.9% in Villa Dolores were obtained. With these findings two endemic areas of systemic mycoses in Cordoba, Argentina were delimited.

  2. Production of an antimicrobial substance against Cryptococcus neoformans by Paenibacillus brasilensis Sa3 isolated from the rhizosphere of Kalanchoe brasiliensis.

    Science.gov (United States)

    Fortes, Tiago Oliveira; Alviano, Daniela Sales; Tupinambá, Gleiser; Padrón, Thaís Souto; Antoniolli, Angelo Roberto; Alviano, Celuta Sales; Seldin, Lucy

    2008-01-01

    An antifungal substance produced by Paenibacillus brasilensis strain Sa3 was preliminary characterized and showed to be stable after treatment with different enzymes and organic solvents and at a wide range of pH, and presented a molecular weight between 3 and 10 kDa. In vitro antagonism of this strain towards Cryptococcus neoformans was investigated by optical and electronic microscopic analyses and a fungicidal effect on C. neoformans was observed. Ultrastructural analysis showed intense changes on the fungus when it was paired cultured with strain Sa3, mainly the detachment of the capsule from the cell wall and the presence of altered organelles in the cytoplasm. This novel antifungal substance produced by P. brasilensis Sa3 may represent a new insight in antifungal therapy mainly against emergent fungi. Also, prospective studies on rhizobacteria of plants as Kalanchoe brasiliensis may offer a potential source for the discovery of bioactive compounds with medical value.

  3. Utilization of fish meal and fish oil for production of Cryptococcus sp. MTCC 5455 lipase and hydrolysis of polyurethane thereof

    OpenAIRE

    Thirunavukarasu, K.; Purushothaman, S.; Gowthaman, M. K.; Nakajima-Kambe, T.; Rose, C.; Kamini, N. R.

    2015-01-01

    Fish meal has been used as an additional nitrogen source and fish oil as inducer for the growth and production of lipase from Cryptococcus sp. MTCC 5455. A response surface design illustrated that the optimum factors influencing lipase production were fish meal, 1.5 %, w/v, Na2HPO4, 0.2 %, w/v, yeast extract, 0.25 %, w/v and sardine oil, 2.0 %, w/v with an activity of 71.23 U/mL at 96 h and 25 °C, which was 48.39 % higher than the conventional one-factor-at-a-time method. The crude concentrat...

  4. Gene Network Polymorphism Illuminates Loss and Retention of Novel RNAi Silencing Components in the Cryptococcus Pathogenic Species Complex.

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    Marianna Feretzaki

    2016-03-01

    Full Text Available RNAi is a ubiquitous pathway that serves central functions throughout eukaryotes, including maintenance of genome stability and repression of transposon expression and movement. However, a number of organisms have lost their RNAi pathways, including the model yeast Saccharomyces cerevisiae, the maize pathogen Ustilago maydis, the human pathogen Cryptococcus deuterogattii, and some human parasite pathogens, suggesting there may be adaptive benefits associated with both retention and loss of RNAi. By comparing the RNAi-deficient genome of the Pacific Northwest Outbreak C. deuterogattii strain R265 with the RNAi-proficient genomes of the Cryptococcus pathogenic species complex, we identified a set of conserved genes that were lost in R265 and all other C. deuterogattii isolates examined. Genetic and molecular analyses reveal several of these lost genes play roles in RNAi pathways. Four novel components were examined further. Znf3 (a zinc finger protein and Qip1 (a homolog of N. crassa Qip were found to be essential for RNAi, while Cpr2 (a constitutive pheromone receptor and Fzc28 (a transcription factor are involved in sex-induced but not mitosis-induced silencing. Our results demonstrate that the mitotic and sex-induced RNAi pathways rely on the same core components, but sex-induced silencing may be a more specific, highly induced variant that involves additional specialized or regulatory components. Our studies further illustrate how gene network polymorphisms involving known components of key cellular pathways can inform identification of novel elements and suggest that RNAi loss may have been a core event in the speciation of C. deuterogattii and possibly contributed to its pathogenic trajectory.

  5. Low diversity Cryptococcus neoformans variety grubii multilocus sequence types from Thailand are consistent with an ancestral African origin.

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    Sitali P Simwami

    2011-04-01

    Full Text Available The global burden of HIV-associated cryptococcal meningitis is estimated at nearly one million cases per year, causing up to a third of all AIDS-related deaths. Molecular epidemiology constitutes the main methodology for understanding the factors underpinning the emergence of this understudied, yet increasingly important, group of pathogenic fungi. Cryptococcus species are notable in the degree that virulence differs amongst lineages, and highly-virulent emerging lineages are changing patterns of human disease both temporally and spatially. Cryptococcus neoformans variety grubii (Cng, serotype A constitutes the most ubiquitous cause of cryptococcal meningitis worldwide, however patterns of molecular diversity are understudied across some regions experiencing significant burdens of disease. We compared 183 clinical and environmental isolates of Cng from one such region, Thailand, Southeast Asia, against a global MLST database of 77 Cng isolates. Population genetic analyses showed that Thailand isolates from 11 provinces were highly homogenous, consisting of the same genetic background (globally known as VNI and exhibiting only ten nearly identical sequence types (STs, with three (STs 44, 45 and 46 dominating our sample. This population contains significantly less diversity when compared against the global population of Cng, specifically Africa. Genetic diversity in Cng was significantly subdivided at the continental level with nearly half (47% of the global STs unique to a genetically diverse and recombining population in Botswana. These patterns of diversity, when combined with evidence from haplotypic networks and coalescent analyses of global populations, are highly suggestive of an expansion of the Cng VNI clade out of Africa, leading to a limited number of genotypes founding the Asian populations. Divergence time testing estimates the time to the most common ancestor between the African and Asian populations to be 6,920 years ago (95% HPD

  6. Scanning electron microscopy of the interaction between Cryptococcus magnus and Colletotrichum gloeosporioides on papaya fruit = Microscopia eletrônica de varredura da interação entre Cryptococcus magnus e Colletotrichum gloeosporioides em frutos de mamão

    NARCIS (Netherlands)

    Capdeville, G.; Souza, M.T.; Santos, J.R.P.; Miranda, S.P.; Caetano, A.R.; Falcao, R.; Gomes, A.C.M.M.

    2007-01-01

    The objective of this work was to investigate possible modes of action of the yeast Cryptococcus magnus in controlling anthracnose (Colletotrichum gloeosporioides) on post harvested papaya fruits. Scanning electron microscopy was used to analyze the effect of the yeast on inoculations done after

  7. Caracterização bioquímica e molecular de Cryptococcus spp. isolados de excretas ambientais de pombos (Columba livia domestica

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    G. Colombo

    2015-12-01

    Full Text Available Os pombos possuem uma rica diversidade de microrganismo, entre eles fungos sapróbios, como do gênero Cryptococcus, que podem atuar como agentes patogênicos para o homem e animais. Objetivou-se o isolamento, a caracterização bioquímica e a molecular de amostras de Cryptococcus spp. de excretas ambientas de pombos. Foram colhidas 100 amostras ambientais de pontos equidistantes e representativos da área da cidade de Araçatuba, São Paulo. As amostras foram rasteladas do solo de vias públicas, armazenadas em frasco coletor e encaminhadas para o Laboratório de Bacteriologia e Micologia da FMVA, onde foram processadas e cultivadas em duplicata, em placas de Petri contendo ágar Sabouraud dextrose a 4% e Niger. Em seguida, foram incubadas à temperatura ambiente e a 30ºC, respectivamente, por um período não inferior a 15 dias. Após a observação diária, as colônias sugestivas para levedura foram reisoladas em ágar Niger e submetidas a testes bioquímicos para posterior caracterização molecular pela técnica da PCR. Como resultado, a caracterização bioquímica e a molecular isolaram 32 colônias leveduriformes, sendo 8% dos cultivos positivos para Cryptococcus neoformans var. neoformans, 17% para Rhodotorula rubidae e 7% Candida albicans. Pelo exposto, concluiu-se que excretas ambientais de pombos constituem um microfoco para Cryptococcus neoformans var. neoformans e outras leveduras com potencial patogênico, representando um risco à saúde pública, sendo necessárias medidas preventivas, como a higienização com a correta remoção das excretas, a fim de minimizar os riscos de exposição ambiental.

  8. Cryptococcus lacticolor sp. nov. and Rhodotorula oligophaga sp. nov., novel yeasts isolated from the nasal smear microbiota of Queensland koalas kept in Japanese zoological parks.

    Science.gov (United States)

    Satoh, Kazuo; Maeda, Mari; Umeda, Yoshiko; Sugamata, Miho; Makimura, Koichi

    2013-07-01

    A total of 515 yeast strains were isolated from the nasal smears of Queensland koalas and their breeding environments in Japanese zoological parks between 2005 and 2012. The most frequent species in the basidiomycetous yeast biota isolated from koala nasal passages was Cryptococcus neoformans, followed by Rhodotorula minuta. R. minuta was the most frequent species in the breeding environments, while C. neoformans was rare. Seven strains representing two novel yeast species were identified. Analyses of the 26S rDNA (LSU) D1/D2 domain and nuclear ribosomal DNA internal transcribed spacer region sequences indicated that these strains represent new species with close phylogenetic relationships to Cryptococcus and Rhodotorula. A sexual state was not found for either of these two novel yeasts. Key phenotypic characters confirmed that these strains could be placed in Cryptococcus and Rhodotorula. The names Cryptococcus lacticolor sp. nov. (type strain TIMM 10013(T) = JCM 15449(T) = CBS 10915(T) = DSM 21093(T), DDBJ/EMBL/Genbank Accession No.; AB375774 (ITS) and AB375775 (26S rDNA D1/D2 region), MycoBank ID; MB 802688, Fungal Barcoding Database ID; 3174), and Rhodotorula oligophaga sp. nov. (type strain TIMM 10017(T) = JCM 18398(T) = CBS 12623(T) = DSM 25814(T), DDBJ/EMBL/Genbank Accession No.; AB702967 (ITS) and AB702967 (26S rDNA D1/D2 region), MycoBank ID; MB 802689, Fungal Barcoding Database ID; 3175) are proposed for these new species.

  9. Cryptococcus Neoformans en excretas de palomas, suelo y aire de los palomares del perímetro Urbano de Ica, 2002

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    Maria Curo

    2005-10-01

    Full Text Available Objetivos: Aislar e identificar las variedades de Cryptococcus neoformans que existen en excretas de palomas, suelo y ambientes aéreos del perímetro urbano de la ciudad de Ica, Perú. Materiales y métodos: Se colectaron muestras de excretas de palomas, suelo contaminado y aire de palomares entre mayo y julio del año 2002. Para el aislamiento primario se usó agar Sabouraud dextrosa con cloramfenicol. Para la identificación por especie se usaron pruebas convencionales y la determinación de la variedad se evaluó sobre el medio de cultivo agar canavanina glicina azul de bromotimol sódico. Resultados: Se obtuvieron 124 muestras procedentes de palomares de la Facultad de Medicina de la Universidad Nacional San Luis Gonzaga de Ica, capilla del Hospital Socorro, Los viñedos de Santa María, La Victoria y San Joaquín. Se aislaron 26 cepas del género Cryptococcus de las cuales nueve cepas correspondieron a C. neoformans var. neoformans y 17 a Cryptococcus spp. La mayor frecuencia se encontró en la zona del palomar de la Facultad de Medicina. Conclusión: C. neoformans var. neoformans se encuentra en excreta y suelo de las áreas protegidas de los palomares estudiados.

  10. Morphologic mimickers of Cryptococcus occurring within inflammatory infiltrates in the setting of neutrophilic dermatitis: a series of three cases highlighting clinical dilemmas associated with a novel histopathologic pitfall.

    Science.gov (United States)

    Ko, Jennifer S; Fernandez, Anthony P; Anderson, Kyle A; Burdick, Laura M; Billings, Steven D; Procop, Gary W; McMahon, James T; Bergfeld, Wilma F; Piliang, Melissa P

    2013-01-01

    A neutrophil-predominant inflammatory infiltrate in a cutaneous biopsy can be associated with a broad spectrum of diseases. Here we describe three cases showing a neutrophil-predominant dermal infiltrate admixed with abundant acellular bodies surrounded by capsule-like vacuolated spaces, which strikingly mimicked Cryptococcus. Two cases occurred within the settings of underlying hematologic malignancies; the third case was associated with immune dysregulation. Two patients were acutely ill in the medical intensive care unit. Fungal work-up, including cultures and multiple stains were negative. Because of clinical deterioration in these patients, transmission electron microscopy was pursued to definitively rule out fungal infection. In both cases, characteristics most compatible with autolysing human cells, not Cryptococcus, were identified. Chemotherapy and high-dose steroids were given, but both patients eventually succumbed to their diseases. To the best of our knowledge, these represent the first reported cases of autolysing human cells mimicking Cryptococcus organisms within neutrophilic infiltrates. They highlight the therapeutic dilemmas arising with histopathologic mimics, as well as the importance of thorough investigation to distinguish mimickers from true infectious organisms. We believe recognition of this microscopic pitfall will be useful to dermatopathologists faced with similar findings in the future, and may prevent unnecessary delay of appropriate therapy in acutely ill patients. Copyright © 2012 John Wiley & Sons A/S.

  11. Prevalência de Cryptococcus neoformans nos pombos urbanos da cidade de Porto Alegre, Rio Grande do Sul Prevalence of Cryptococcus neoformans in urban pigeons of Porto Alegre (RS, Brazil

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    Aline Reolon

    2004-10-01

    Full Text Available Cryptococcus neoformans, levedura encapsulada, é o agente etiológico da criptococose em humanos e animais. A variedade neoformans, encontrada em fontes ambientais, incluindo fezes de pombos, é importante causa de mortalidade em indivíduos imunodeprimidos em todo o mundo. Contudo, ainda não há estudos sobre sua ecologia na cidade de Porto Alegre, onde se tem registro da ocorrência de casos humanos dessa micose. Para pesquisar fontes saprofíticas de C. neoformans na cidade de Porto Alegre, foram coletadas 88 amostras de excretas de pombos em distintas praças da cidade. Suspensões das amostras em salina estéril foram semeadas em placas com meio ágar Sabouraud e incubadas em estufa a temperaturas entre 29 e 30,5°C. Após cinco dias, colônias mucóides foram subcultivadas para identificação através de provas morfofisiológicas. O fungo C. neoformans foi isolado em 88 (100% das amostras avaliadas, confirmando sua ocorrência nos pombos que habitam as praças da cidade.Cryptococcus neoformans is an encapsulated yeast, causing cryptococcosis in human beings and animals. The variety neoformans, found in environmental sources, including pigeon excrements, is an important cause of mortality in immunocompromised individuals in the whole world. However, there are still no studies on its ecology in the city of Porto Alegre, where register of the occurrence of human cases of this mycosis is found. To study the saprophitic sources of C. neoformans in the city of Porto Alegre, 88 samples of avian excreta have been collected in distinct squares. Suspension of the samples in sterile saline solution had been planted in plates with Sabouraud agar and incubated in stove at 29-30.5°C. After five days mucoid colonies were subcultived for identification through morphologic and physiologic tests. C. neoformans was isolated in 88 (100% of the samples, proving its occurrence in the pigeons that inhabit the squares of that city.

  12. Comparative genomics of Cryptococcus neoformans var. grubii associated with meningitis in HIV infected and uninfected patients in Vietnam.

    Science.gov (United States)

    Day, Jeremy N; Qihui, Seet; Thanh, Lam Tuan; Trieu, Phan Hai; Van, Anh Duong; Thu, Nha Hoang; Chau, Tran Thi Hong; Lan, Nguyen P H; Chau, Nguyen Van Vinh; Ashton, Philip M; Thwaites, Guy E; Boni, Maciej F; Wolbers, Marcel; Nagarajan, Niranjan; Tan, Patrick B O; Baker, Stephen

    2017-06-01

    The vast burden of cryptococcal meningitis occurs in immunosuppressed patients, driven by HIV, and is caused by Cryptococcus neoformans var. grubii. We previously reported cryptococcal meningitis in Vietnam arising atypically in HIV uninfected, apparently immunocompetent patients, caused by a single amplified fragment length polymorphism (AFLP) cluster of C. neoformans var. grubii (VNIγ). This variant was less common in HIV infected individuals; it remains unclear why this lineage is associated with apparently immunocompetent patients. To study this host tropism we aimed to further our understanding of clinical phenotype and genomic variation within Vietnamese C. neoformans var. grubii. After performing MLST on C. neoformans clinical isolates we identified 14 sequence types (STs); ST5 correlated with the VNIγ cluster. We next compared clinical phenotype by lineage and found HIV infected patients with cryptococcal meningitis caused by ST5 organisms were significantly more likely to have lymphadenopathy (11% vs. 4%, p = 0.05 Fisher's exact test) and higher blood lymphocyte count (median 0.76 versus 0.55 X109 cells/L, p = 0.001, Kruskal-Wallis test). Furthermore, survivors of ST5 infections had evidence of worse disability outcomes at 70 days (72.7% (40/55) in ST5 infections versus 57.1% (52/91) non-ST5 infections (OR 2.11, 95%CI 1.01 to 4.41), p = 0.046). To further investigate the relationship between strain and disease phenotype we performed genome sequencing on eight Vietnamese C. neoformans var. grubii. Eight genome assemblies exhibited >99% nucleotide sequence identity and we identified 165 kbp of lineage specific to Vietnamese isolates. ST5 genomes harbored several strain specific regions, incorporating 19 annotated coding sequences and eight hypothetical proteins. These regions included a phenolic acid decarboxylase, a DEAD-box ATP-dependent RNA helicase 26, oxoprolinases, a taurine catabolism dioxygenase, a zinc finger protein, membrane transport proteins

  13. Phialemonium Fungemia: Two Documented Nosocomial Cases

    OpenAIRE

    Guarro, Josep; Nucci, Marcio; Akiti, Tiyomi; Gené, Josepa; Cano, Josep; Barreiro, M. Da Gloria C.; Aguilar, Carme

    1999-01-01

    Two fungal isolates recovered from the blood of two immunosuppressed patients are described as Phialemonium curvatum. One patient died, while the other, who was infected with Exophiala jeanselmei at the same time, survived after successful treatment with itraconazole. Analysis of internal transcribed spacer sequences demonstrated that the isolates belonged to the same strain and that the source of infection was probably a catheter. The taxonomic position of P. curvatum is discussed, and Phial...

  14. Cativeiro de aves como fonte de Cryptococcus neoformans na cidade de Campo Grande, Mato Grosso do Sul, Brasil

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    Filiú Wander Fernando de Oliveira

    2002-01-01

    Full Text Available Cryptococcus neoformans é a levedura capsulada causadora de criptococose em humanos e animais. A variedade neoformans, encontrada em diversas fontes ambientais, inclusive habitats de aves, é importante causa de mortalidade em indivíduos com AIDS em todo o Mundo. Contudo, ainda não há estudos sobre a sua ecologia na região Centro Oeste brasileira, onde há registro da ocorrência de casos humanos da micose. Para estudar fontes saprofíticas de C. neoformans, na cidade de Campo Grande, foram coletadas 20 amostras de excretas de aves em distintos ambientes. Suspensão das amostras em salina estéril foram semeadas em placas com meio ágar níger. Após 5 dias, colônias mucóides marrom-escuro foram subcultivadas para identificação através de provas morfofisiológicas, determinação da variedade e sorotipagem. C. neoformans var. neoformans sorotipo A foi isolado de 10 (50% das amostras, comprovando a ocorrência saprofítica de C. neoformans na cidade de Campo Grande, relacionada a habitat de aves em cativeiro.

  15. A ‘suicide’ CRISPR-Cas9 system to promote gene deletion and restoration by electroporation in Cryptococcus neoformans

    Science.gov (United States)

    Wang, Yu; Wei, Dongsheng; Zhu, Xiangyang; Pan, Jiao; Zhang, Ping; Huo, Liang; Zhu, Xudong

    2016-01-01

    Loss-of-function mutagenesis is an important tool used to characterize gene functions, and the CRISPR-Cas9 system is a powerful method for performing targeted mutagenesis in organisms that present low recombination frequencies, such as the serotype D strains of Cryptococcus neoformans. However, when the CRISPR-Cas9 system persists in the host cells, off-target effects and Cas9 cytotoxicity may occur, which might block subsequent genetic manipulation. Here, we report a method of spontaneously eliminating the CRISPR-Cas9 system without impairing its robust editing function. We successfully expressed single guide RNA under the driver of an endogenous U6 promoter and the human codon-optimized Cas9 endonuclease with an ACT1 promoter. This system can effectively generate an indel mutation and efficiently perform targeted gene disruption via homology-directed repair by electroporation in yeast. We then demonstrated the spontaneous elimination of the system via a cis arrangement of the CRISPR-Cas9 expression cassettes to the recombination construct. After a system-mediated double crossover, the CRISPR-Cas9 cassettes were cleaved and degraded, which was validated by Southern blotting. This ‘suicide’ CRISPR-Cas9 system enables the validation of gene functions by subsequent complementation and has the potential to minimize off-target effects. Thus, this technique has the potential for use in functional genomics studies of C. neoformans. PMID:27503169

  16. Fostering triacylglycerol accumulation in novel oleaginous yeast Cryptococcus psychrotolerans IITRFD utilizing groundnut shell for improved biodiesel production.

    Science.gov (United States)

    Deeba, Farha; Pruthi, Vikas; Negi, Yuvraj S

    2017-10-01

    The investigation was carried out to examine the potential of triacylglycerol (TAG) accumulation by novel oleaginous yeast isolate Cryptococcus psychrotolerans IITRFD on utilizing groundnut shell acid hydrolysate (GSH) as cost-effective medium. The maximum biomass productivity and lipid productivity of 0.095±0.008g/L/h and 0.044±0.005g/L/h, respectively with lipid content 46% was recorded on GSH. Fatty acid methyl ester (FAME) profile obtained by GC-MS analysis revealed oleic acid (37.8%), palmitic (29.4%) and linoleic (32.8%) as major fatty acids representing balance between oxidative stability (OS) and cold flow filter properties (CFFP) for improved biodiesel quality. The biodiesel property calculated were correlated well with the fuel standards limits of ASTM D6751, EN 14214 and IS 15607. The present findings raise the possibility of using agricultural waste groundnut shell as a substrate for production of biodiesel by novel oleaginous yeast isolate C. psychrotolerans IITRFD. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. First report of urease activity in the novel systemic fungal pathogen Emergomyces africanus: a comparison with the neurotrope Cryptococcus neoformans.

    Science.gov (United States)

    Lerm, Barbra; Kenyon, Chris; Schwartz, Ilan S; Kroukamp, Heinrich; de Witt, Riaan; Govender, Nelesh P; de Hoog, G Sybren; Botha, Alfred

    2017-11-01

    Cryptococcus neoformans is an opportunistic pathogen responsible for the AIDS-defining illness, cryptococcal meningitis. During the disease process, entry of cryptococcal cells into the brain is facilitated by virulence factors that include urease enzyme activity. A novel species of an Emmonsia-like fungus, recently named Emergomyces africanus, was identified as a cause of disseminated mycosis in HIV-infected persons in South Africa. However, in contrast to C. neoformans, the enzymes produced by this fungus, some of which may be involved in pathogenesis, have not been described. Using a clinical isolate of C. neoformans as a reference, the study aim was to confirm, characterise and quantify urease activity in E. africanus clinical isolates. Urease activity was tested using Christensen's urea agar, after which the presence of a urease gene in the genome of E. africanus was confirmed using gene sequence analysis. Subsequent evaluation of colorimetric enzyme assay data, using Michaelis-Menten enzyme kinetics, revealed similarities between the substrate affinity of the urease enzyme produced by E. africanus (Km ca. 26.0 mM) and that of C. neoformans (Km ca. 20.6 mM). However, the addition of 2.5 g/l urea to the culture medium stimulated urease activity of E. africanus, whereas nutrient limitation notably increased cryptococcal urease activity. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Identification of genes from the fungal pathogen Cryptococcus neoformans related to transmigration into the central nervous system.

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    Hsiang-Kuang Tseng

    Full Text Available A mouse brain transmigration assessment (MBTA was created to investigate the central nervous system (CNS pathogenesis of cryptococcal meningoencephalitis.Two cryptococcal mutants were identified from a pool of 109 pre-selected mutants that were signature-tagged with the nourseothricin acetyltransferase (NAT resistance cassette. These two mutants displayed abnormal transmigration into the central nervous system. One mutant displaying decreased transmigration contains a null mutation in the putative FNX1 gene, whereas the other mutant possessing a null mutation in the putative RUB1 gene exhibited increased transmigration into the brain. Two macrophage adhesion-defective mutants in the pool, 12F1 and 3C9, showed reduced phagocytosis by macrophages, but displayed no defects in CNS entry suggesting that transit within macrophages (the "Trojan horse" model of CNS entry is not the primary mechanism for C. neoformans migration into the CNS in this MBTA.This research design provides a new strategy for genetic impact studies on how Cryptococcus passes through the blood-brain barrier (BBB, and the specific isolated mutants in this assay support a transcellular mechanism of CNS entry.

  19. A link between virulence and homeostatic responses to hypoxia during infection by the human fungal pathogen Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Cheryl D Chun

    2007-02-01

    Full Text Available Fungal pathogens of humans require molecular oxygen for several essential biochemical reactions, yet virtually nothing is known about how they adapt to the relatively hypoxic environment of infected tissues. We isolated mutants defective in growth under hypoxic conditions, but normal for growth in normoxic conditions, in Cryptococcus neoformans, the most common cause of fungal meningitis. Two regulatory pathways were identified: one homologous to the mammalian sterol-response element binding protein (SREBP cholesterol biosynthesis regulatory pathway, and the other a two-component-like pathway involving a fungal-specific hybrid histidine kinase family member, Tco1. We show that cleavage of the SREBP precursor homolog Sre1-which is predicted to release its DNA-binding domain from the membrane-occurs in response to hypoxia, and that Sre1 is required for hypoxic induction of genes encoding for oxygen-dependent enzymes involved in ergosterol synthesis. Importantly, mutants in either the SREBP pathway or the Tco1 pathway display defects in their ability to proliferate in host tissues and to cause disease in infected mice, linking for the first time to our knowledge hypoxic adaptation and pathogenesis by a eukaryotic aerobe. SREBP pathway mutants were found to be a hundred times more sensitive than wild-type to fluconazole, a widely used antifungal agent that inhibits ergosterol synthesis, suggesting that inhibitors of SREBP processing could substantially enhance the potency of current therapies.

  20. IL-4 receptor-alpha-dependent control of Cryptococcus neoformans in the early phase of pulmonary infection.

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    Andreas Grahnert

    Full Text Available Cryptococcus neoformans is an opportunistic fungal pathogen that causes lung inflammation and meningoencephalitis in immunocompromised people. Previously we showed that mice succumb to intranasal infection by induction of pulmonary interleukin (IL-4Rα-dependent type 2 immune responses, whereas IL-12-dependent type 1 responses confer resistance. In the experiments presented here, IL-4Rα⁻/⁻ mice unexpectedly show decreased fungal control early upon infection with C. neoformans, whereas wild-type mice are able to control fungal growth accompanied by enhanced macrophage and dendritic cell recruitment to the site of infection. Lower pulmonary recruitment of macrophages and dendritic cells in IL-4Rα⁻/⁻ mice is associated with reduced pulmonary expression of CCL2 and CCL20 chemokines. Moreover, IFN-γ and nitric oxide production are diminished in IL-4Rα⁻/⁻ mice compared to wild-type mice. To directly study the potential mechanism(s responsible for reduced production of IFN-γ, conventional dendritic cells were stimulated with C. neoformans in the presence of IL-4 which results in increased IL-12 production and reduced IL-10 production. Together, a beneficial role of early IL-4Rα signaling is demonstrated in pulmonary cryptococcosis, which contrasts with the well-known IL-4Rα-mediated detrimental effects in the late phase.

  1. Opsonization of Cryptococcus neoformans by a family of isotype-switch variant antibodies specific for the capsular polysaccharide.

    Science.gov (United States)

    Schlageter, A M; Kozel, T R

    1990-06-01

    A family of immunoglobulin isotype-switch variants was isolated by sib selection from a murine hybridoma which produced an immunoglobulin G subclass 1 (IgG1) antibody specific for the capsular polysaccharide of Cryptococcus neoformans. Antibodies of the IgG1, IgG2a, and IgG2b isotypes had similar serotype specificity patterns in double immunodiffusion assays which used polysaccharides of the four cryptococcal serotypes as antigens. A quantitative difference in the ability of the isotypes to form a precipitate with the polysaccharide was observed in a double immunodiffusion assay and confirmed in a quantitative precipitin assay. The relative precipitating activity of the antibodies was IgG2a greater than IgG1 much greater than IgG2b. Analysis by enzyme-linked immunosorbent assay of the reactivity of the three isotypes with cryptococcal polysaccharide showed identical titers and slopes, suggesting that the variable region of the class-switch antibodies was unaltered. This system allowed us to examine the effect of the Fc portion of the antibody on opsonization of encapsulated cryptococci. Yeast cells were precoated with antibodies of each isotype and incubated with murine macrophages or cultured human monocytes. Antibodies of all three isotypes exhibited a dose-dependent opsonization for phagocytosis by both human and murine phagocytes. The relative opsonic activity of the antibodies was IgG2a greater than IgG1 greater than IgG2b.

  2. Microsatellite typing of clinical and environmental Cryptococcus neoformans var. grubii isolates from Cuba shows multiple genetic lineages.

    Directory of Open Access Journals (Sweden)

    Maria-Teresa Illnait-Zaragozi

    Full Text Available BACKGROUND: Human cryptococcal infections have been associated with bird droppings as a likely source of infection. Studies toward the local and global epidemiology of Cryptococcus spp. have been hampered by the lack of rapid, discriminatory, and exchangeable molecular typing methods. METHODOLOGY/PRINCIPAL FINDINGS: We selected nine microsatellite markers for high-resolution fingerprinting from the genome of C. neoformans var. grubii. This panel of markers was applied to a collection of clinical (n = 122 and environmental (n = 68; from pigeon guano C. neoformans var. grubii isolates from Cuba. All markers proved to be polymorphic. The average number of alleles per marker was 9 (range 5-51. A total of 104 genotypes could be distinguished. The discriminatory power of this panel of markers was 0.993. Multiple clusters of related genotypes could be discriminated that differed in only one or two microsatellite markers. These clusters were assigned as microsatellite complexes. The majority of environmental isolates (>70% fell into 1 microsatellite complex containing only few clinical isolates (49 environmental versus 2 clinical. Clinical isolates were segregated over multiple microsatellite complexes. CONCLUSIONS/SIGNIFICANCE: A large genotypic variation exists in C. neoformans var. grubii. The genotypic segregation between clinical and environmental isolates from pigeon guano suggests additional source(s of human cryptococcal infections. The selected panel of microsatellite markers is an excellent tool to study the epidemiology of C. neoformans var. grubii.

  3. A Unique Fungal Two-Component System Regulates Stress Responses, Drug Sensitivity, Sexual Development, and Virulence of Cryptococcus neoformans

    Science.gov (United States)

    Bahn, Yong-Sun; Kojima, Kaihei; Cox, Gary M.

    2006-01-01

    The stress-activated mitogen-activated protein kinase (MAPK) pathway is widely used by eukaryotic organisms as a central conduit via which cellular responses to the environment effect growth and differentiation. The basidiomycetous human fungal pathogen Cryptococcus neoformans uniquely uses the stress-activated Pbs2-Hog1 MAPK system to govern a plethora of cellular events, including stress responses, drug sensitivity, sexual reproduction, and virulence. Here, we characterized a fungal “two-component” system that controls these fundamental cellular functions via the Pbs2-Hog1 MAPK cascade. A typical response regulator, Ssk1, modulated all Hog1-dependent phenotypes by controlling Hog1 phosphorylation, indicating that Ssk1 is the major upstream signaling component of the Pbs2-Hog1 pathway. A second response regulator, Skn7, governs sensitivity to Na+ ions and the antifungal agent fludioxonil, negatively controls melanin production, and functions independently of Hog1 regulation. To control these response regulators, C. neoformans uses multiple sensor kinases, including two-component–like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. Our findings highlight unique adaptations of this global two-component MAPK signaling cascade in a ubiquitous human fungal pathogen. PMID:16672377

  4. Utilization of fish meal and fish oil for production of Cryptococcus sp. MTCC 5455 lipase and hydrolysis of polyurethane thereof.

    Science.gov (United States)

    Thirunavukarasu, K; Purushothaman, S; Gowthaman, M K; Nakajima-Kambe, T; Rose, C; Kamini, N R

    2015-09-01

    Fish meal has been used as an additional nitrogen source and fish oil as inducer for the growth and production of lipase from Cryptococcus sp. MTCC 5455. A response surface design illustrated that the optimum factors influencing lipase production were fish meal, 1.5 %, w/v, Na2HPO4, 0.2 %, w/v, yeast extract, 0.25 %, w/v and sardine oil, 2.0 %, w/v with an activity of 71.23 U/mL at 96 h and 25 °C, which was 48.39 % higher than the conventional one-factor-at-a-time method. The crude concentrated enzyme hydrolyzed polyurethane (PUR) efficiently and hydrolysis was 94 % at 30 °C and 96 h. The products, diethylene glycol and adipic acid were quantified by HPLC and scanning electron microscopic studies of the degraded polymer showed significant increase in size of the holes from 24 to 72 h of incubation. Hydrolysis of PUR within 96 h makes the lipase novel for disposal of PUR and provides an innovative solution to the problems created by plastic wastes.

  5. Vacuolar zinc transporter Zrc1 is required for detoxification of excess intracellular zinc in the human fungal pathogen Cryptococcus neoformans.

    Science.gov (United States)

    Cho, Minsu; Hu, Guanggan; Caza, Mélissa; Horianopoulos, Linda C; Kronstad, James W; Jung, Won Hee

    2018-01-01

    Zinc is an important transition metal in all living organisms and is required for numerous biological processes. However, excess zinc can also be toxic to cells and cause cellular stress. In the model fungus Saccharomyces cerevisiae, a vacuolar zinc transporter, Zrc1, plays important roles in the storage and detoxification of excess intracellular zinc to protect the cell. In this study, we identified an ortholog of the S. cerevisiae ZRC1 gene in the human fungal pathogen Cryptococcus neoformans. Zrc1 was localized in the vacuolar membrane in C. neoformans, and a mutant lacking ZRC1 showed significant growth defects under high-zinc conditions. These results suggested a role for Zrc1 in zinc detoxification. However, contrary to our expectation, the expression of Zrc1 was induced in cells grown in zinc-limited conditions and decreased upon the addition of zinc. These expression patterns were similar to those of Zip1, the high-affinity zinc transporter in the plasma membrane of C. neoformans. Furthermore, we used the zrc1 mutant in a murine model of cryptococcosis to examine whether a mammalian host could inhibit the survival of C. neoformans using zinc toxicity. We found that the mutant showed no difference in virulence compared with the wildtype strain. This result suggests that Zrc1-mediated zinc detoxification is not required for the virulence of C. neoformans, and imply that zinc toxicity may not be an important aspect of the host immune response to the fungus.

  6. PRM1 and KAR5 function in cell-cell fusion and karyogamy to drive distinct bisexual and unisexual cycles in the Cryptococcus pathogenic species complex.

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    Ci Fu

    2017-11-01

    Full Text Available Sexual reproduction is critical for successful evolution of eukaryotic organisms in adaptation to changing environments. In the opportunistic human fungal pathogens, the Cryptococcus pathogenic species complex, C. neoformans primarily undergoes bisexual reproduction, while C. deneoformans undergoes both unisexual and bisexual reproduction. During both unisexual and bisexual cycles, a common set of genetic circuits regulates a yeast-to-hyphal morphological transition, that produces either monokaryotic or dikaryotic hyphae. As such, both the unisexual and bisexual cycles can generate genotypic and phenotypic diversity de novo. Despite the similarities between these two cycles, genetic and morphological differences exist, such as the absence of an opposite mating-type partner and monokaryotic instead of dikaryotic hyphae during C. deneoformans unisexual cycle. To better understand the similarities and differences between these modes of sexual reproduction, we focused on two cellular processes involved in sexual reproduction: cell-cell fusion and karyogamy. We identified orthologs of the plasma membrane fusion protein Prm1 and the nuclear membrane fusion protein Kar5 in both Cryptococcus species, and demonstrated their conserved roles in cell fusion and karyogamy during C. deneoformans α-α unisexual reproduction and C. deneoformans and C. neoformans a-α bisexual reproduction. Notably, karyogamy occurs inside the basidum during bisexual reproduction in C. neoformans, but often occurs earlier following cell fusion during bisexual reproduction in C. deneoformans. Characterization of these two genes also showed that cell fusion is dispensable for solo unisexual reproduction in C. deneoformans. The blastospores produced along hyphae during C. deneoformans unisexual reproduction are diploid, suggesting that diploidization occurs early during hyphal development, possibly through either an endoreplication pathway or cell fusion-independent karyogamy

  7. Identification of a major IP5 kinase in Cryptococcus neoformans confirms that PP-IP5/IP7, not IP6, is essential for virulence

    OpenAIRE

    Li, Cecilia; Lev, Sophie; Saiardi, Adolfo; Desmarini, Desmarini; Sorrell, Tania C.; Djordjevic, Julianne T.

    2016-01-01

    Fungal inositol polyphosphate (IP) kinases catalyse phosphorylation of IP3 to inositol pyrophosphate, PP-IP5/IP7, which is essential for virulence of Cryptococcus neoformans. Cryptococcal Kcs1 converts IP6 to PP-IP5/IP7, but the kinase converting IP5 to IP6 is unknown. Deletion of a putative IP5 kinase-encoding gene (IPK1) alone (ipk1?), and in combination with KCS1 (ipk1?kcs1?), profoundly reduced virulence in mice. However, deletion of KCS1 and IPK1 had a greater impact on virulence attenua...

  8. Repression of Proteases and Hsp90 Chaperone Expression Induced by an Antiretroviral in Virulent Environmental Strains of Cryptococcus neoformans.

    Science.gov (United States)

    Serafin, Cleber Fernando; Paris, Ana Paula; Paula, Claudete Rodrigues; Simão, Rita Cássia Garcia; Gandra, Rinaldo Ferreira

    2017-04-01

    This study evaluated the effect of the antiretroviral ritonavir on protease secretion in different strains of Cryptococcus neoformans isolated from the environment and investigated the expression of heat shock protein (Hsp90), classically described virulence factors in other yeast in the presence of the same antiretroviral. The presence of the enzyme was detected by the formation of a degradation of the halo around the colonies. The results were classified as follows: level 1 (without proteases), level 2 (positive for proteases), and level 3 (strongly positive for proteases). Total protein extract isolated from the cell walls of the 12 strains incubated in the absence and presence of ritonavir (0.3125 mg mL -1 ) were resolved by SDS-PAGE and analyzed by Western blot assays using an antiserum against Hsp90 from Blastocladiella emersonii. All strains tested showed inhibition of proteinase activity in the presence of ritonavir at 0.3125 to 1.25 mg mL -1 . High levels of Hsp90 were observed in the absence of ritonavir (0.3125 mg mL -1 ), except for the non-virulent control cells. In contrast, in the presence of the antiretroviral, a drastic reduction in the expression of the chaperone was observed. The data suggest that ritonavir, in addition to containing viral replication, could inhibit the expression of virulence factors in opportunistic yeast, as proteases and Hsp90. According to our current knowledge, this is the first time that the inhibition of Hsp90 by an antiretroviral was reported for environmental isolates of C. neoformans.

  9. Characterization of additional components of the environmental pH-sensing complex in the pathogenic fungus Cryptococcus neoformans.

    Science.gov (United States)

    Pianalto, Kaila M; Ost, Kyla S; Brown, Hannah E; Alspaugh, J Andrew

    2018-05-16

    Pathogenic microorganisms must adapt to changes in their immediate surroundings, including alterations in pH, to survive the shift from the external environment to that of the infected host. In the basidiomycete fungal pathogen Cryptococcus neoformans , these pH changes are primarily sensed by the fungal-specific, alkaline pH-sensing Rim/Pal pathway. The C. neoformans Rim pathway has diverged significantly from that described in ascomycete fungi. We recently identified the C. neoformans putative pH sensor Rra1, which activates the Rim pathway in response to elevated pH. In this study, we probed the function of Rra1 by analyzing its cellular localization and performing protein co-immunoprecipitation to identify potential Rra1 interactors. We found that Rra1 does not strongly colocalize or interact with immediate downstream Rim pathway components. However, these experiments identified a novel Rra1 interactor, the previously uncharacterized C. neoformans nucleosome assembly protein 1 (Nap1), which was required for Rim pathway activation. We observed that Nap1 specifically binds to the C-terminal tail of the Rra1 sensor, likely promoting Rra1 protein stability. This function of Nap1 is conserved in fungi closely related to C. neoformans that contain Rra1 orthologs, but not in the more distantly-related ascomycete fungus Saccharomyces cerevisiae In conclusion, our findings have revealed the sophisticated, yet distinct, molecular mechanisms by which closely and distantly related microbial phyla rapidly adapt to environmental signals and changes such as alterations in pH. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Protection of melanized Cryptococcus neoformans from lethal dose gamma irradiation involves changes in melanin's chemical structure and paramagnetism.

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    Abdelahad Khajo

    Full Text Available Certain fungi thrive in highly radioactive environments including the defunct Chernobyl nuclear reactor. Cryptococcus neoformans (C. neoformans, which uses L-3,4-dihydroxyphenylalanine (L-DOPA to produce melanin, was used here to investigate how gamma radiation under aqueous aerobic conditions affects the properties of melanin, with the aim of gaining insight into its radioprotective role. Exposure of melanized fungal cell in aqueous suspensions to doses of γ-radiation capable of killing 50 to 80% of the cells did not lead to a detectable loss of melanin integrity according to EPR spectra of melanin radicals. Moreover, upon UV-visible (Xe-lamp illumination of melanized cells, the increase in radical population was unchanged after γ-irradiation. Gamma-irradiation of frozen cell suspensions and storage of samples for several days at 77 K however, produced melanin modification noted by a reduced radical population and reduced photoresponse. More direct evidence for structural modification of melanin came from the detection of soluble products with absorbance maxima near 260 nm in supernatants collected after γ-irradiation of cells and cell-free melanin. These products, which include thiobarbituric acid (TBA-reactive aldehydes, were also generated by Fenton reagent treatment of cells and cell-free melanin. In an assay of melanin integrity based on the metal (Bi(+3 binding capacity of cells, no detectable loss in binding was detected after γ-irradiation. Our results show that melanin in C. neoformans cells is susceptible to some damage by hydroxyl radical formed in lethal radioactive aqueous environments and serves a protective role in melanized fungi that involves sacrificial breakdown.

  11. Binding of purified and radioiodinated capsular polysaccharides from Cryptococcus neoformans serotype A strains to capsule-free mutants

    International Nuclear Information System (INIS)

    Small, J.M.; Mitchell, T.G.

    1986-01-01

    Strains 6, 15, 98, 110, and 145 of Cryptococcus neoformans serotype A vary in capsule size, animal virulence, and susceptibility to in vitro phagocytosis. The isolated capsular polysaccharides (CPSs) differ in monosaccharide composition ratios and molecular size, as determined by gel filtration. The purpose of this investigation was to characterize the binding of CPSs to capsule-free mutants of C. neoformans and to examine CPSs from these strains for differences in their ability to bind, to determine whether such differences might explain the variation in the pathobiology of these strains. CPSs were partially periodate oxidized, tyraminated, iodinated with 125 I, and used in binding studies with two capsule-free mutants of C. neoformans, strain 602 and Cap59. Binding was specific for yeast species and for polysaccharide and was saturable, which is consistent with a receptor-mediated mechanism of attachment. Binding occurred rapidly and was only slowly reversible. Binding was also independent of pH from pH 5.5 to 8, of cation concentrations, and of competition by sugars up to 1.0 M concentrations. Only a portion of CPS was capable of binding, and strains varied in the extent to which their CPS bound. CPS-15-IV (peak IV was the major polysaccharide peak on DEAE-cellulose chromatography of CPS from strain 15) had the highest proportion of binding (40%), followed by CPS from strains 98, 6, 145, 110, and 15-III (peak III was an earlier eluting fraction of CPS from strain 15). The CPSs differed similarly in their ability to competitively inhibit binding. Treatment of CPS, but not yeast cells, with proteinase XIV abolished binding without altering the CPS gross structure. Treatment of yeast cells with proteases, heat, or formaldehyde did not alter binding, and both strain 602 and Cap59 bound CPS similarly. Binding to encapsulated yeast cells was minimal

  12. Solid-state NMR Reveals the Carbon-based Molecular Architecture of Cryptococcus neoformans Fungal Eumelanins in the Cell Wall*

    Science.gov (United States)

    Chatterjee, Subhasish; Prados-Rosales, Rafael; Itin, Boris; Casadevall, Arturo; Stark, Ruth E.

    2015-01-01

    Melanin pigments protect against both ionizing radiation and free radicals and have potential soil remediation capabilities. Eumelanins produced by pathogenic Cryptococcus neoformans fungi are virulence factors that render the fungal cells resistant to host defenses and certain antifungal drugs. Because of their insoluble and amorphous characteristics, neither the pigment bonding framework nor the cellular interactions underlying melanization of C. neoformans have yielded to comprehensive molecular-scale investigation. This study used the C. neoformans requirement of exogenous obligatory catecholamine precursors for melanization to produce isotopically enriched pigment “ghosts” and applied 2D 13C-13C correlation solid-state NMR to reveal the carbon-based architecture of intact natural eumelanin assemblies in fungal cells. We demonstrated that the aliphatic moieties of solid C. neoformans melanin ghosts include cell-wall components derived from polysaccharides and/or chitin that are associated proximally with lipid membrane constituents. Prior to development of the mature aromatic fungal pigment, these aliphatic moieties form a chemically resistant framework that could serve as the scaffold for melanin synthesis. The indole-based core aromatic moieties show interconnections that are consistent with proposed melanin structures consisting of stacked planar assemblies, which are associated spatially with the aliphatic scaffold. The pyrrole aromatic carbons of the pigments bind covalently to the aliphatic framework via glycoside or glyceride functional groups. These findings establish that the structure of the pigment assembly changes with time and provide the first biophysical information on the mechanism by which melanin is assembled in the fungal cell wall, offering vital insights that can advance the design of bioinspired conductive nanomaterials and novel therapeutics. PMID:25825492

  13. Solid-state NMR Reveals the Carbon-based Molecular Architecture of Cryptococcus neoformans Fungal Eumelanins in the Cell Wall.

    Science.gov (United States)

    Chatterjee, Subhasish; Prados-Rosales, Rafael; Itin, Boris; Casadevall, Arturo; Stark, Ruth E

    2015-05-29

    Melanin pigments protect against both ionizing radiation and free radicals and have potential soil remediation capabilities. Eumelanins produced by pathogenic Cryptococcus neoformans fungi are virulence factors that render the fungal cells resistant to host defenses and certain antifungal drugs. Because of their insoluble and amorphous characteristics, neither the pigment bonding framework nor the cellular interactions underlying melanization of C. neoformans have yielded to comprehensive molecular-scale investigation. This study used the C. neoformans requirement of exogenous obligatory catecholamine precursors for melanization to produce isotopically enriched pigment "ghosts" and applied 2D (13)C-(13)C correlation solid-state NMR to reveal the carbon-based architecture of intact natural eumelanin assemblies in fungal cells. We demonstrated that the aliphatic moieties of solid C. neoformans melanin ghosts include cell-wall components derived from polysaccharides and/or chitin that are associated proximally with lipid membrane constituents. Prior to development of the mature aromatic fungal pigment, these aliphatic moieties form a chemically resistant framework that could serve as the scaffold for melanin synthesis. The indole-based core aromatic moieties show interconnections that are consistent with proposed melanin structures consisting of stacked planar assemblies, which are associated spatially with the aliphatic scaffold. The pyrrole aromatic carbons of the pigments bind covalently to the aliphatic framework via glycoside or glyceride functional groups. These findings establish that the structure of the pigment assembly changes with time and provide the first biophysical information on the mechanism by which melanin is assembled in the fungal cell wall, offering vital insights that can advance the design of bioinspired conductive nanomaterials and novel therapeutics. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  14. SEC14 is a specific requirement for secretion of phospholipase B1 and pathogenicity of Cryptococcus neoformans

    Science.gov (United States)

    Chayakulkeeree, Methee; Johnston, Simon Andrew; Oei, Johanes Bijosono; Lev, Sophie; Williamson, Peter Richard; Wilson, Christabel Frewen; Zuo, Xiaoming; Leal, Ana Lusia; Vainstein, Marilene Henning; Meyer, Wieland; Sorrell, Tania Christine; May, Robin Charles; Djordjevic, Julianne Teresa

    2011-01-01

    Summary Secreted phospholipase B1 (CnPlb1) is essential for dissemination of Cryptococcus neoformans to the central nervous system (CNS) yet essential components of its secretion machinery remain to be elucidated. Using gene deletion analysis we demonstrate that CnPlb1 secretion is dependent on the CnSEC14 product, CnSec14-1p. CnSec14-1p is a homologue of the phosphatidylinositol transfer protein (PITP) ScSec14p, which is essential for secretion and viability in Saccharomyces cerevisiae. In contrast to CnPlb1, neither laccase 1 (Lac1)-induced melanization within the cell wall nor capsule induction were negatively impacted in CnSEC14-1 deletion mutants (CnΔsec14-1 and CnΔsec14-1CnΔsfh5). Similar to the CnPLB1 deletion mutant (CnΔplb1), CnΔsec14-1 was hypo-virulent in mice and did not disseminate to the CNS by day 14 post infection. Furthermore, macrophage expulsion of live CnΔsec14-1 and CnΔplb1 (vomocytosis) was reduced. Individual deletion of CnSEC14-2, a closely-related CnSEC14-1 homologue, and CnSFH5, a distantly-related SEC fourteen-like homologue, did not abrogate CnPlb1 secretion or virulence. However, reconstitution of CnΔsec14-1 with CnSEC14-1 or CnSEC14-2 restored both phenotypes, consistent with functional genetic redundancy. We conclude that CnPlb1 secretion is SEC14-dependent and that C. neoformans preferentially exports virulence determinants to the cell periphery via distinct pathways. We also demonstrate that CnPlb1 secretion is essential for vomocytosis. PMID:21453402

  15. Passive immunization against Cryptococcus neoformans with an isotype-switch family of monoclonal antibodies reactive with cryptococcal polysaccharide.

    Science.gov (United States)

    Sanford, J E; Lupan, D M; Schlageter, A M; Kozel, T R

    1990-01-01

    The in vivo properties of an immunoglobulin isotype-switch family of monoclonal antibodies specific for the polysaccharide capsule of Cryptococcus neoformans were examined in a murine model of cryptococcosis. Subclass-switch variants were isolated by sequential sublining of an immunoglobulin G subclass 1 (IgG1)-secreting cell line. Antibodies of the IgG1, IgG2a, and IgG2b isotypes with identical reactivities with cryptococcal polysaccharide were prepared. The antibodies had the distinct biological properties associated with the heavy chains of each respective isotype. The antibodies were used prophylactically or therapeutically in an attempt to alter the course of cryptococcal infection in mice. Survival of mice and a tissue census of the numbers of viable cryptococci in the lung, spleen, and brain were used as indicators of efficacy. Passive immunization with the IgG2a and IgG2b antibodies effected a reduction in the numbers of cryptococci in lung and spleen. Passive immunization with the IgG1 antibody was markedly less effective. Passive immunization had little or no effect on the numbers of cryptococci in brain tissue, regardless of the immunoglobulin isotype. Despite apparent efficacy with regard to reduction in the numbers of yeast cells in the lung and spleen, the results showed no improvement in survival from murine cryptococcosis. Our results indicate that passive immunization produces a modest effect on the course of murine cryptococcosis in tissues other than brain. However, under the experimental conditions used, such treatment does not have a measurable impact on the ultimate outcome of the infection. PMID:2341184

  16. Contribution of IL-1RI Signaling to Protection against Cryptococcus neoformans 52D in a Mouse Model of Infection

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    Mitra Shourian

    2018-01-01

    Full Text Available Interleukin-1 alpha (IL-1α and interleukin-1 beta (IL-1β are pro-inflammatory cytokines that are induced after Cryptococcus neoformans infection and activate the interleukin-1 receptor type I (IL-1RI. To establish the role of IL-1RI signaling in protection against cryptococcal infection, we analyzed wild-type (WT and IL-1RI-deficient (IL-1RI−/− mice on the BALB/c background. IL-1RI−/− mice had significantly reduced survival compared to WT mice after intratracheal challenge with C. neoformans 52D. Microbiological analysis showed a significant increase in the lung and brain fungal burden of IL-1RI−/− compared to WT mice beginning at weeks 1 and 4 postinfection, respectively. Histopathology showed that IL-1RI−/− mice exhibit greater airway epithelial mucus secretion and prominent eosinophilic crystals that were absent in WT mice. Susceptibility of IL-1RI−/− mice was associated with significant induction of a Th2-biased immune response characterized by pulmonary eosinophilia, M2 macrophage polarization, and recruitment of CD4+ IL-13+ T cells. Expression of pro-inflammatory [IL-1α, IL-1β, TNFα, and monocyte chemoattractant protein 1 (MCP-1], Th1-associated (IFNγ, and Th17-associated (IL-17A cytokines was significantly reduced in IL-1RI−/− lungs compared to WT. WT mice also had higher expression of KC/CXCL1 and sustained neutrophil recruitment to the lung; however, antibody-mediated depletion of these cells showed that they were dispensable for lung fungal clearance. In conclusion, our data indicate that IL-1RI signaling is required to activate a complex series of innate and adaptive immune responses that collectively enhance host defense and survival after C. neoformans 52D infection in BALB/c mice.

  17. Unisexual and heterosexual meiotic reproduction generate aneuploidy and phenotypic diversity de novo in the yeast Cryptococcus neoformans.

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    Min Ni

    2013-09-01

    Full Text Available Aneuploidy is known to be deleterious and underlies several common human diseases, including cancer and genetic disorders such as trisomy 21 in Down's syndrome. In contrast, aneuploidy can also be advantageous and in fungi confers antifungal drug resistance and enables rapid adaptive evolution. We report here that sexual reproduction generates phenotypic and genotypic diversity in the human pathogenic yeast Cryptococcus neoformans, which is globally distributed and commonly infects individuals with compromised immunity, such as HIV/AIDS patients, causing life-threatening meningoencephalitis. C. neoformans has a defined a-α opposite sexual cycle; however, >99% of isolates are of the α mating type. Interestingly, α cells can undergo α-α unisexual reproduction, even involving genotypically identical cells. A central question is: Why would cells mate with themselves given that sex is costly and typically serves to admix preexisting genetic diversity from genetically divergent parents? In this study, we demonstrate that α-α unisexual reproduction frequently generates phenotypic diversity, and the majority of these variant progeny are aneuploid. Aneuploidy is responsible for the observed phenotypic changes, as chromosome loss restoring euploidy results in a wild-type phenotype. Other genetic changes, including diploidization, chromosome length polymorphisms, SNPs, and indels, were also generated. Phenotypic/genotypic changes were not observed following asexual mitotic reproduction. Aneuploidy was also detected in progeny from a-α opposite-sex congenic mating; thus, both homothallic and heterothallic sexual reproduction can generate phenotypic diversity de novo. Our study suggests that the ability to undergo unisexual reproduction may be an evolutionary strategy for eukaryotic microbial pathogens, enabling de novo genotypic and phenotypic plasticity and facilitating rapid adaptation to novel environments.

  18. Insights into the mechanisms of protective immunity against Cryptococcus neoformans infection using a mouse model of pulmonary cryptococcosis.

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    Karen L Wozniak

    2009-09-01

    Full Text Available Cryptococcus neoformans is an opportunistic fungal pathogen that causes life-threatening pneumonia and meningoencephalitis in immune compromised individuals. Previous studies have shown that immunization of BALB/c mice with an IFN-gamma-producing C. neoformans strain, H99gamma, results in complete protection against a second pulmonary challenge with an otherwise lethal cryptococcal strain. The current study evaluated local anamnestic cell-mediated immune responses against pulmonary cryptococcosis in mice immunized with C. neoformans strain H99gamma compared to mice immunized with heat-killed C. neoformans (HKC.n.. Mice immunized with C. neoformans strain H99gamma had significantly reduced pulmonary fungal burden post-secondary challenge compared to mice immunized with HKC.n. Protection against pulmonary cryptococcosis was associated with increased pulmonary granulomatous formation and leukocyte infiltration followed by a rapid resolution of pulmonary inflammation, which protected the lungs from severe allergic bronchopulmonary mycosis (ABPM-pathology that developed in the lungs of mice immunized with HKC.n. Pulmonary challenge of interleukin (IL-4 receptor, IL-12p40, IL-12p35, IFN-gamma, T cell and B cell deficient mice with C. neoformans strain H99gamma demonstrated a requirement for Th1-type T cell-mediated immunity, but not B cell-mediated immunity, for the induction of H99gamma-mediated protective immune responses against pulmonary C. neoformans infection. CD4(+ T cells, CD11c(+ cells, and Gr-1(+ cells were increased in both proportion and absolute number in protected mice. In addition, significantly increased production of Th1-type/pro-inflammatory cytokines and chemokines, and conversely, reduced Th2-type cytokine production was observed in the lungs of protected mice. Interestingly, protection was not associated with increased production of cytokines IFN-gamma or TNF-alpha in lungs of protected mice. In conclusion, immunization with C

  19. Deteccion de antigeno del polisacarido capsular de Cryptococcus neoformans en pacientes con SIDA y neurocriptococosis en São Paulo, Brasil

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    B. Calvo

    1991-12-01

    Full Text Available Antígeno del polisacárido capsular (Ag PC de Cryptococcus neoformans fue detectado por la técnica de aglutinación de latex (AL en LCR y suero de pacientes con Sindrome de Inmunodeficiencia Adquirida (SIDA y primer episodio de neurocriptococosis, usando como patrón el examen micológico (examen directo y cultivo de LCR. Se obtuvo una sensibilidad del 100% de AL para detectar AgPC de C. neoformans, el cual por su rápidez permite tratamiento específico precoz. Títulos iniciales de AgPC de la levadura en esos pacientes pueden ser > 1.000.000, pareciendo que cuando esos títulos están presentes en suero, se relacionan con mortalidad durante el tratamiento. En los pacientes que sobrevivieron se observó que el examen micológico directo y AgPC de C. neoformans, en LCR y suero, permanecen positivos aún después de tratamiento y mejoría clínica del paciente.Capsular polysaccharide antigen (AgPC of Cryptococcus neoformans was detected by latex agglutination technique (LA in cerebrospinal fluid and serum of patients with AIDS during their first central nervous system manifestation of the disease. Direct mycological examination and culture were used as controls. Sensitivity was 100% by LA allowing an early specific treatment of cryptococcosis. Initial titres of AgPC in such patients can be > 1000000 and it appears that when such titres are present in the serum they are related to mortality during treatment. Surviving patients showed positivity of the direct mycological examination and AgPC of C. neoformans in cerebrospinal fluid and sera even after treatment and clinical recovery.

  20. Pilzinfektionen des Zentralnervensystems bei immunkompetentem Wirt

    NARCIS (Netherlands)

    Tintelnot, K.; de Hoog, G.S.; Haase, G.

    2014-01-01

    Die Mehrzahl von Pilzinfektionen, die zu mykotischen Tumoren führen und bei Patienten ohne jegliche prädisponierende Vorerkrankung auftreten, wird durch Cryptococcus gattii bzw. C. neoformans oder durch Schwärzepilze, insbesondere durch Cladophialophora bantiana, Ramichloridium mackenziei, Exophiala

  1. Spread of Rare Fungus from Vancouver Island

    Centers for Disease Control (CDC) Podcasts

    2006-12-20

    Cryptococcus gattii, a rare fungus normally found in the tropics, has infected people and animals on Vancouver Island, Canada. Dr. David Warnock, Director, Division of Foodborne, Bacterial, and Mycotic Diseases, CDC, discusses public health concerns about further spread of this organism.  Created: 12/20/2006 by Emerging Infectious Diseases.   Date Released: 12/29/2006.

  2. Spread of Rare Fungus from Vancouver Island

    Centers for Disease Control (CDC) Podcasts

    Cryptococcus gattii, a rare fungus normally found in the tropics, has infected people and animals on Vancouver Island, Canada. Dr. David Warnock, Director, Division of Foodborne, Bacterial, and Mycotic Diseases, CDC, discusses public health concerns about further spread of this organism

  3. A glycosylphosphatidylinositol anchor is required for membrane localization but dispensable for cell wall association of chitin deacetylase 2 in Cryptococcus neoformans.

    Science.gov (United States)

    Gilbert, Nicole M; Baker, Lorina G; Specht, Charles A; Lodge, Jennifer K

    2012-01-01

    Cell wall proteins (CWPs) mediate important cellular processes in fungi, including adhesion, invasion, biofilm formation, and flocculation. The current model of fungal cell wall organization includes a major class of CWPs covalently bound to β-1,6-glucan via a remnant of a glycosylphosphatidylinositol (GPI) anchor. This model was established by studies of ascomycetes more than a decade ago, and relatively little work has been done with other fungi, although the presumption has been that proteins identified in the cell wall which contain a predicted GPI anchor are covalently linked to cell wall glucans. The pathogenic basidiomycete Cryptococcus neoformans encodes >50 putatively GPI-anchored proteins, some of which have been identified in the cell wall. One of these proteins is chitin deacetylase 2 (Cda2), an enzyme responsible for converting chitin to chitosan, a cell wall polymer recently established as a virulence factor for C. neoformans infection of mammalian hosts. Using a combination of biochemistry, molecular biology, and genetics, we show that Cda2 is GPI anchored to membranes but noncovalently associated with the cell wall by means independent of both its GPI anchor and β-1,6-glucan. We also show that Cda2 produces chitosan when localized to the plasma membrane, but association with the cell wall is not essential for this process, thereby providing insight into the mechanism of chitosan biosynthesis. These results increase our understanding of the surface of C. neoformans and provide models of cell walls likely applicable to other undercharacterized basidiomycete pathogenic fungi. The surface of a pathogenic microbe is a major interface with its host. In fungi, the outer surface consists of a complex matrix known as the cell wall, which includes polysaccharides, proteins, and other molecules. The mammalian host recognizes many of these surface molecules and mounts appropriate responses to combat the microbial infection. Cryptococcus neoformans is a

  4. Scanning electron microscopy of the interaction between Cryptococcus magnus and Colletotrichum gloeosporioides on papaya fruit Microscopia eletrônica de varredura da interação entre Cryptococcus magnus e Colletotrichum gloeosporioides em frutos de mamão

    Directory of Open Access Journals (Sweden)

    Guy de Capdeville

    2007-11-01

    Full Text Available The objective of this work was to investigate possible modes of action of the yeast Cryptococcus magnus in controlling anthracnose (Colletotrichum gloeosporioides on post harvested papaya fruits. Scanning electron microscopy was used to analyze the effect of the yeast on inoculations done after harvest. Results showed that C. magnus is able to colonize wound surfaces much faster than the pathogen, outcompeting the later for space and probably for nutrients. In addition, C. magnus produces a flocculent matrix, which affects hyphae integrity. The competition for space and the production of substances that affect hyphae integrity are among the most important modes of action of this yeast.O objetivo deste trabalho foi investigar prováveis modos de ação da levedura Cryptococcus magnus, que resultam no controle da antracnose (Colletotrichum gloeosporioides em frutos de mamoeiro na póscolheita. A microscopia eletrônica de varredura foi utilizada para avaliar o efeito da levedura sobre inoculações realizadas após a colheita. Os resultados mostraram que C. magnus é capaz de colonizar a superfície de ferimentos nos frutos e vencer a competição por espaço e, provavelmente, por nutrientes. Além disso, C. magnus produz uma matriz de textura característica que afeta a integridade da hifa do patógeno. A competição por espaço e a produção de substâncias que afetam a integridade das hifas estão entre os mais importantes modos de ação desta levedura.

  5. Illuminating choices for library prep: a comparison of library preparation methods for whole genome sequencing of Cryptococcus neoformans using Illumina HiSeq.

    Directory of Open Access Journals (Sweden)

    Johanna Rhodes

    Full Text Available The industry of next-generation sequencing is constantly evolving, with novel library preparation methods and new sequencing machines being released by the major sequencing technology companies annually. The Illumina TruSeq v2 library preparation method was the most widely used kit and the market leader; however, it has now been discontinued, and in 2013 was replaced by the TruSeq Nano and TruSeq PCR-free methods, leaving a gap in knowledge regarding which is the most appropriate library preparation method to use. Here, we used isolates from the pathogenic fungi Cryptococcus neoformans var. grubii and sequenced them using the existing TruSeq DNA v2 kit (Illumina, along with two new kits: the TruSeq Nano DNA kit (Illumina and the NEBNext Ultra DNA kit (New England Biolabs to provide a comparison. Compared to the original TruSeq DNA v2 kit, both newer kits gave equivalent or better sequencing data, with increased coverage. When comparing the two newer kits, we found little difference in cost and workflow, with the NEBNext Ultra both slightly cheaper and faster than the TruSeq Nano. However, the quality of data generated using the TruSeq Nano DNA kit was superior due to higher coverage at regions of low GC content, and more SNPs identified. Researchers should therefore evaluate their resources and the type of application (and hence data quality being considered when ultimately deciding on which library prep method to use.

  6. Increases thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase by fusion of cellulose binding domain derived from Trichoderma reesei

    International Nuclear Information System (INIS)

    Thongekkaew, Jantaporn; Ikeda, Hiroko; Iefuji, Haruyuki

    2012-01-01

    Highlights: ► The CSLP and fusion enzyme were successfully expressed in the Pichia pastoris. ► The fusion enzyme was stable at 80 °C for 120-min. ► The fusion enzyme was responsible for cellulose-binding capacity. ► The fusion enzyme has an attractive applicant for enzyme immobilization. -- Abstract: To improve the thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase (CSLP), the cellulose-binding domain originates from Trichoderma reesei cellobiohydrolase I was engineered into C-terminal region of the CSLP (CSLP-CBD). The CSLP and CSLP-CBD were successfully expressed in the Pichia pastoris using the strong methanol inducible alcohol oxidase 1 (AOX1) promoter and the secretion signal sequence from Saccharomyces cerevisiae (α factor). The recombinant CSLP and CSLP-CBD were secreted into culture medium and estimated by SDS–PAGE to be 22 and 27 kDa, respectively. The fusion enzyme was stable at 80 °C and retained more than 80% of its activity after 120-min incubation at this temperature. Our results also found that the fusion of fungal exoglucanase cellulose-binding domain to CSLP is responsible for cellulose-binding capacity. This attribute should make it an attractive applicant for enzyme immobilization.

  7. Characterization of sophorolipid biosurfactant produced by Cryptococcus sp. VITGBN2 and its application on Zn(II) removal from electroplating wastewater.

    Science.gov (United States)

    Basak, Geetanjali; Das, Nilanjana

    2014-11-01

    The present study aimed at elucidating the role of biosurfactant produced by yeast for the removal of Zn(II) ions from electroplating wastewater. The yeast species isolated from CETP, Vellore, Tamilnadu was identified as Cryptococcus sp.VITGBN2, based on molecular techniques, and was found to be potent producer of biosurfactant in mineral salt media containing vegetable oil as additional carbon source. Chemical structure of the purified biosurfactant was identified as acidic diacetate sophorolipid through GC-MS analysis. Interaction of Zn(II) ions with biosurfactant was monitored using FT-IR, SEM and EDS analysis. Zn (II) removal at 100 mg l(-1) concentration was 84.8% compared were other synthetic surfactants (Tween 80 and sodium dodecyl sulphate), yeast mediated biosurfactant showed enhanced Zn (II) removal in batch mode. The role of biosurfactant on Zn(II) removal was evaluated in column mode packed with biosurfactant entrapped in sodium alginate beads. At a flow rate of 1 ml min(-1) and bed height of 12 cm, immobilized biosurfactant showed 94.34% Zn(II) removal from electroplating wastewater. The present study confirmed that Zn(II) removal was biosurfactant mediated. This is the first report establishing the involvement of yeast mediated biosurfactant in Zn(II) removal from wastewater.

  8. Analysis of the Protein Kinase A-Regulated Proteome of Cryptococcus neoformans Identifies a Role for the Ubiquitin-Proteasome Pathway in Capsule Formation

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    J. M. H. Geddes

    2016-01-01

    Full Text Available The opportunistic fungal pathogen Cryptococcus neoformans causes life-threatening meningitis in immunocompromised individuals. The expression of virulence factors, including capsule and melanin, is in part regulated by the cyclic-AMP/protein kinase A (cAMP/PKA signal transduction pathway. In this study, we investigated the influence of PKA on the composition of the intracellular proteome to obtain a comprehensive understanding of the regulation that underpins virulence. Through quantitative proteomics, enrichment and bioinformatic analyses, and an interactome study, we uncovered a pattern of PKA regulation for proteins associated with translation, the proteasome, metabolism, amino acid biosynthesis, and virulence-related functions. PKA regulation of the ubiquitin-proteasome pathway in C. neoformans showed a striking parallel with connections between PKA and protein degradation in chronic neurodegenerative disorders and other human diseases. Further investigation of proteasome function with the inhibitor bortezomib revealed an impact on capsule production as well as hypersusceptibility for strains with altered expression or activity of PKA. Parallel studies with tunicamycin also linked endoplasmic reticulum stress with capsule production and PKA. Taken together, the data suggest a model whereby expression of PKA regulatory and catalytic subunits and the activation of PKA influence proteostasis and the function of the endoplasmic reticulum to control the elaboration of the polysaccharide capsule. Overall, this study revealed both broad and conserved influences of the cAMP/PKA pathway on the proteome and identified proteostasis as a potential therapeutic target for the treatment of cryptococcosis.

  9. Increases thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase by fusion of cellulose binding domain derived from Trichoderma reesei

    Energy Technology Data Exchange (ETDEWEB)

    Thongekkaew, Jantaporn, E-mail: jantaporn_25@yahoo.com [Department of Biological Science, Faculty of Science, Ubon-Ratchathani University, Warinchumrab, Ubon-Ratchathani 34190 (Thailand); Ikeda, Hiroko; Iefuji, Haruyuki [Application Research Division, National Research Institute of Brewing, 3-7-1 Kagamiyama, Higashi-Hiroshima 739-0046 (Japan)

    2012-03-30

    Highlights: Black-Right-Pointing-Pointer The CSLP and fusion enzyme were successfully expressed in the Pichia pastoris. Black-Right-Pointing-Pointer The fusion enzyme was stable at 80 Degree-Sign C for 120-min. Black-Right-Pointing-Pointer The fusion enzyme was responsible for cellulose-binding capacity. Black-Right-Pointing-Pointer The fusion enzyme has an attractive applicant for enzyme immobilization. -- Abstract: To improve the thermal stability and cellulose-binding capacity of Cryptococcus sp. S-2 lipase (CSLP), the cellulose-binding domain originates from Trichoderma reesei cellobiohydrolase I was engineered into C-terminal region of the CSLP (CSLP-CBD). The CSLP and CSLP-CBD were successfully expressed in the Pichia pastoris using the strong methanol inducible alcohol oxidase 1 (AOX1) promoter and the secretion signal sequence from Saccharomyces cerevisiae ({alpha} factor). The recombinant CSLP and CSLP-CBD were secreted into culture medium and estimated by SDS-PAGE to be 22 and 27 kDa, respectively. The fusion enzyme was stable at 80 Degree-Sign C and retained more than 80% of its activity after 120-min incubation at this temperature. Our results also found that the fusion of fungal exoglucanase cellulose-binding domain to CSLP is responsible for cellulose-binding capacity. This attribute should make it an attractive applicant for enzyme immobilization.

  10. The Southeastern Asian house mouse (Mus musculus castaneus Linn.) as a new passenger host for Cryptococcus neoformans var. grubii molecular type VNI.

    Science.gov (United States)

    Singh, Karuna; Rani, Jyoti; Neelabh; Rai, Govind Kumar; Singh, Major

    2017-11-01

    We describe Mus musculus castaneus as a new mammalian host for Cryptococcus neoformans var. grubii (VNI). Eighteen apparently healthy adults and pups of the rodent were collected from human dwellings in Varanasi, a city of India. Both clinical and behavioral examinations of the rodents did not reveal any sign of the disease. Among visceral organs, histological examination of only liver exhibited the presence of single celled, encapsulated, Southgate's mucicarmine positive fungal structures consistent with C. neoformans. Nevertheless, culture of tissue homogenates of brain, lungs, liver, and kidneys yielded white colonies on Sabouraud's dextrose agar and brown mucoid colonies of C. neoformans on Staib's and Tobacco agar media. The pathogen was isolated from habitat soil as well as fresh faeces of the animals. All isolates were urease positive, nitrate and canavanine-glycine bromothymol blue negative, exhibited phenoloxidase activity and grew at 37°C. The isolates were identified as C. neoformans var. grubii with ITS primers and unique marker (GACA)4. The pathogen when inoculated in immunosuppressed mice showed low pathogenicity. To our knowledge, we for the first time report case cluster of Mus musculus castaneus as new passenger host for C. neoformans var. grubii (VNI). © The Author 2017. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Illuminating choices for library prep: a comparison of library preparation methods for whole genome sequencing of Cryptococcus neoformans using Illumina HiSeq.

    Science.gov (United States)

    Rhodes, Johanna; Beale, Mathew A; Fisher, Matthew C

    2014-01-01

    The industry of next-generation sequencing is constantly evolving, with novel library preparation methods and new sequencing machines being released by the major sequencing technology companies annually. The Illumina TruSeq v2 library preparation method was the most widely used kit and the market leader; however, it has now been discontinued, and in 2013 was replaced by the TruSeq Nano and TruSeq PCR-free methods, leaving a gap in knowledge regarding which is the most appropriate library preparation method to use. Here, we used isolates from the pathogenic fungi Cryptococcus neoformans var. grubii and sequenced them using the existing TruSeq DNA v2 kit (Illumina), along with two new kits: the TruSeq Nano DNA kit (Illumina) and the NEBNext Ultra DNA kit (New England Biolabs) to provide a comparison. Compared to the original TruSeq DNA v2 kit, both newer kits gave equivalent or better sequencing data, with increased coverage. When comparing the two newer kits, we found little difference in cost and workflow, with the NEBNext Ultra both slightly cheaper and faster than the TruSeq Nano. However, the quality of data generated using the TruSeq Nano DNA kit was superior due to higher coverage at regions of low GC content, and more SNPs identified. Researchers should therefore evaluate their resources and the type of application (and hence data quality) being considered when ultimately deciding on which library prep method to use.

  12. Perfil bioquímico do soro de frangos de corte alimentados com dieta suplementada com alfa-amilase de Cryptococcus flavus e Aspergillus niger HM2003 Biochemichal serum profile of broilers fed diets suplemented with alfa-amylase from Cryptococcus flavus and Aspergillus niger HM2003

    Directory of Open Access Journals (Sweden)

    Cibele Silva Minafra

    2010-12-01

    Full Text Available Avaliou-se o perfil bioquímico do soro de frangos de corte alimentados com a enzima α-amilase produzida por dois microrganismos. Produziram-se dois extratos, um com a-amilase obtida a partir de Cryptococcus flavus em meio de levedura comercial e outro com Aspergillus niger HM2003 em meio de proteína de soja e amido comercial, com atividade de 9,58 U/mL e 10,0 U/mL, respectivamente. Utilizaram-se 360 pintos de corte Cobb 500 de 1 dia de idade e com 49,72 ± 0,68 g de peso vivo inicial. As aves foram alojadas em baterias e foram criadas até os 21 dias de idade. Foram utilizados três dietas, cada uma com cinco repetições de 12 aves, em delineamento inteiramente casualizado. A primeira dieta (basal foi formulada sem adição de enzima e as outras duas receberam a suplementação de a-amilase produzida por cultivo de Cryptococcus flavus e Aspergillus niger HM2003. Dietas à base de milho e soja foram formuladas em duas fases: pré-inicial (1-7 dias e inicial (8-21 dias. Na fase pré-inicial, foram observados os seguintes valores médios para cálcio (6,90 e 5,99 mg/dL, proteína plasmática (2,0 e 2,50 g/dL e fosfatase alcalina (979,98 e 974,66 UI/L, respectivamente para Cryptococcus flavus e Aspergillus niger HM2003. A dieta acrescida de a-amilase obtida a partir de Aspergillus niger HM2003 determinou maior concentração sérica de fósforo. Na fase inicial, os resultados significativos relacionaram-se a potássio quando avaliadas dietas com adição de a-amilase pelas duas fontes. A incorporação das enzimas testadas não proporciona alterações metabólicas ou toxicidade nos animais.It was evaluated the biochemical serum profile of broilers fed rations supplemented with α-amylase produced by two microorganisms. Two extracts were produced, one was produced with a-amylase obtained from Cryptococcus flavus in a commercial yeast-based medium and the other with Aspergillus niger HM2003 produced in soybean protein and commercial starch medium

  13. Antifungal susceptibilities of Candida, Cryptococcus neoformans and Aspergillus fumigatus from the Asia and Western Pacific region: data from the SENTRY antifungal surveillance program (2010-2012).

    Science.gov (United States)

    Pfaller, Michael A; Messer, Shawn A; Jones, Ronald N; Castanheira, Mariana

    2015-09-01

    The SENTRY Antifungal Surveillance Program monitors global susceptibility rates of newer and established antifungal agents. We report the in vitro activity of seven antifungal agents against 496 contemporary clinical isolates of yeasts and molds. The isolates were obtained from 20 laboratories in the Asia-Western Pacific (APAC) region during 2010 through 2012. Anidulafungin, caspofungin, micafungin, fluconazole, itraconazole, posaconazole and voriconazole were susceptibility tested using CLSI methods and species-specific interpretive criteria. Sequencing of fks hot spots was performed for echinocandin-resistant strains. Isolates included 13 species of Candida (n=460), 5 species of non-Candida yeasts (21), 5 species of Aspergillus (11) and 4 other molds. Echinocandin resistance was uncommon among eight species of Candida and was only detected in three isolates of Candida glabrata, two from Australia harboring mutations in fks1 (F625S) and fks2 (S663P). Resistance to the azoles was much more common and was observed among all species with the exception of Candida dubliniensis. Fluconazole resistance rates observed with C. glabrata (6.8%) was comparable to that seen with Candida parapsilosis (5.7%) and Candida tropicalis (3.6%). Cross resistance among the triazoles was seen with each of these three species. The mold-active azoles and the echinocandins were all active against isolates of Aspergillus fumigatus. Azole resistance was not detected among the isolates of Cryptococcus neoformans. Antifungal resistance is uncommon among isolates of fungi causing invasive fungal infections in the APAC region. As in other regions of the world, emerging resistance to the echinocandins among invasive isolates of C. glabrata bears close monitoring.

  14. First report of environmental isolation of Cryptococcus neoformans and other fungi from pigeon droppings in Makkah, Saudi Arabia and in vitro susceptibility testing

    Directory of Open Access Journals (Sweden)

    Hussein Hasan Abulreesh

    2015-08-01

    Full Text Available Objective: To verify the occurrence of Cryptococcus neoformans (C. neoformans and other fungi in samples of pigeon droppings collected from Makkah city, Saudi Arabia. Methods: One hundred and twelve withered pigeon dropping samples were collected from 12 different districts. Using the dilution plate technique, samples were cultured on Sabouraud dextrose agar and esculin agar. Colonies were examined microscopically and C. neoformans identification is confirmed by India ink preparation, observation of urease activity and brown pigmentation on esculin medium. Susceptibility patterns of five yeast species and four molds against five antifungal drugs were tested using agar disk diffusion method. Results: C. neoformans was recovered from 38 samples (34%. Na'aman valley was recorded to be the highest contaminated site (66.7% with C. neoformans, while the samples collected from Al Awaly district were considered as the lowest contaminated samples (6.7%. Also, twenty species related to sixteen genera of fungi other than C. neoformans were recovered from which, three yeast genera were recorded. The antifungal susceptibility testing showed that the nine tested fungal species were sensitive to Mycosat, while Fungican exerted inhibition zones of four species only. C. neoformans was moderately sensitive towards all tested compounds but it can resist Flucoral where no inhibition zone could be detected. Conclusions: Our results are considered to be the first report on the environmental prevalence of C. neoformans in pigeon feces in Makkah, Saudi Arabia. The data indicated that pigeon droppings can be considered as a potential source of this basidiomycetous yeast in addition to other fungal species in this region.

  15. The production of monokaryotic hyphae by Cryptococcus neoformans can be induced by high temperature arrest of the cell cycle and is independent of same-sex mating.

    Science.gov (United States)

    Fu, Jianmin; Morris, Ian R; Wickes, Brian L

    2013-01-01

    Cryptococcus neoformans is a heterothallic fungal pathogen of humans and animals. Although the fungus grows primarily as a yeast, hyphae are produced during the sexual phase and during a process called monokaryotic fruiting, which is also believed to involve sexual reproduction, but between cells of the same mating type. Here we report a novel monokaryotic fruiting mechanism that is dependent on the cell cycle and occurs in haploid cells in the absence of sexual reproduction. Cells grown at 37°C were found to rapidly produce hyphae (∼4 hrs) and at high frequency (∼40% of the population) after inoculation onto hyphae-inducing agar. Microscopic examination of the 37°C seed culture revealed a mixture of normal-sized and enlarged cells. Micromanipulation of single cells demonstrated that only enlarged cells were able to produce hyphae and genetic analysis confirmed that hyphae did not arise from α-α mating or endoduplication. Cell cycle analysis revealed that cells grown at 37°C had an increased population of cells in G2 arrest, with the proportion correlated with the frequency of monokaryotic fruiting. Cell sorting experiments demonstrated that enlarged cells were only found in the G2-arrested population and only this population contained cells able to produce hyphae. Treatment of cells at low temperature with the G2 cell cycle arrest agent, nocodazole, induced hyphal growth, confirming the role of the cell cycle in this process. Taken together, these results reveal a mating-independent mechanism for monokaryotic fruiting, which is dependent on the cell cycle for induction of hyphal competency.

  16. Two cation transporters Ena1 and Nha1 cooperatively modulate ion homeostasis, antifungal drug resistance, and virulence of Cryptococcus neoformans via the HOG pathway

    Science.gov (United States)

    Jung, Kwang-Woo; Strain, Anna K; Nielsen, Kirsten; Jung, Kwang-Hwan; Bahn, Yong-Sun

    2012-01-01

    Maintenance of cation homeostasis is essential for survival of all living organisms in their biological niches. It is also important for the survival of human pathogenic fungi in the host, where cation concentrations and pH will vary depending on different anatomical sites. However, the exact role of diverse cation transporters and ion channels in virulence of fungal pathogens remains elusive. In this study we functionally characterized ENA1 and NHA1, encoding a putative Na+/ATPase and Na+/H+ antiporter, respectively, in Cryptococcus neoformans, a basidiomycete fungal pathogen which causes fatal meningoencephalitis. Expression of NHA1 and ENA1 is induced in response to salt and osmotic shock mainly in a Hog1-dependent manner. Phenotypic analysis of the ena1, nha1, and ena1 nha1 mutants revealed that Ena1 controls cellular levels of toxic cations, such as Na+ and Li+ whereas both Ena1 and Nha1 are important for controlling less toxic K+ ions. Under alkaline conditions, Ena1 was highly induced and required for growth in the presence of low levels of Na+ or K+ salt and Nha1 played a role in survival under K+ stress. In contrast, Nha1, but not Ena1, was essential for survival at acidic conditions (pH 4.5) under high K+ stress. In addition, Ena1 and Nha1 were required for maintenance of plasma membrane potential and stability, which appeared to modulate antifungal drug susceptibility. Perturbation of ENA1 and NHA1 enhanced capsule production and melanin synthesis. However, Nha1 was dispensable for virulence of C. neoformans although Ena1 was essential. In conclusion, Ena1 and Nha1 play redundant and discrete roles in cation homeostasis, pH regulation, membrane potential, and virulence in C. neoformans, suggesting that these transporters could be novel antifungal drug targets for treatment of cryptococcosis. PMID:22343280

  17. Activation and binding of opsonic fragments of C3 on encapsulated Cryptococcus neoformans by using an alternative complement pathway reconstituted from six isolated proteins.

    Science.gov (United States)

    Kozel, T R; Wilson, M A; Pfrommer, G S; Schlageter, A M

    1989-07-01

    Encapsulated Cryptococcus neoformans yeast cells are potent activators of the complement system. We examined the interaction of the yeast cells with an alternative complement pathway reconstituted from isolated factor D, factor B, factor H, factor I, C3, and properdin. Incubation of encapsulated cryptococci with the reconstituted pathway led to activation and binding of C3 fragments to the yeast cells that was quantitatively and qualitatively identical to that observed with normal human serum. Incubation with either normal serum or a mixture of isolated proteins led to binding of 4 x 10(7) to 5 x 10(7) C3 molecules to the yeast cells. The kinetics for activation and binding of C3 were identical, with maximum binding observed after a 20-min incubation. Immunoglobulin G was not needed for optimal activation kinetics. C3 fragments eluted from the yeast cells by treatment with hydroxylamine and subsequent analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated the presence primarily of iC3b on yeast cells incubated with either normal serum or the reconstituted pathway. Ultrastructural examination of the opsonized yeast cells showed that the cryptococcal capsule was the site for binding of C3 activated from normal serum or the reconstituted pathway, with a dense accumulation of C3 at the periphery of the capsule. Thus, incubation of encapsulated cryptococci in the reconstituted pathway led to deposition of opsonic complement fragments at a site that was appropriate for interaction with phagocyte receptors. Cryptococci opsonized with the reconstituted pathway showed a markedly enhanced interaction with cultured human monocytes compared with unopsonized yeast cells, indicating that the alternative pathway alone is opsonic for yeast cells. However, the results indicate that additional serum factors are needed for optimal opsonization of yeast cells because a 35% reduction in the number of cryptococci bound to macrophages was observed with

  18. Genome-Wide Identification of circRNAs in Pathogenic Basidiomycetous Yeast Cryptococcus neoformans Suggests Conserved circRNA Host Genes over Kingdoms

    Directory of Open Access Journals (Sweden)

    Liang Huo

    2018-02-01

    Full Text Available Circular RNAs (circRNAs, a novel class of ubiquitous and intriguing noncoding RNA, have been found in a number of eukaryotes but not yet basidiomycetes. In this study, we identified 73 circRNAs from 39.28 million filtered RNA reads from the basidiomycete Cryptococcus neoformans JEC21 using next-generation sequencing (NGS and the bioinformatics tool circular RNA identification (CIRI. Furthermore, mapping of newly found circRNAs to the genome showed that 73.97% of the circRNAs originated from exonic regions, whereas 20.55% were from intergenic regions and 5.48% were from intronic regions. Enrichment analysis of circRNA host genes was conducted based on the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway databases. The results reveal that host genes are mainly responsible for primary metabolism and, interestingly, ribosomal protein production. Furthermore, we uncovered a high-level circRNA that was a transcript from the guanosine triphosphate (GTPase gene CNM01190 (gene ID: 3255052 in our yeast. Coincidentally, YPT5, CNM01190′s ortholog of the GTPase in Schizosaccharomyces pombe, protists, and humans, has already been proven to generate circRNAs. Additionally, overexpression of RNA debranching enzyme DBR1 had varied influence on the expression of circRNAs, indicating that multiple circRNA biosynthesis pathways exist in C. neoformans. Our study provides evidence for the existence of stable circRNAs in the opportunistic human pathogen C. neoformans and raises a question regarding their role related to pathogenesis in this yeast.

  19. Rat eosinophils stimulate the expansion of Cryptococcus neoformans-specific CD4+ and CD8+ T cells with a T-helper 1 profile

    Science.gov (United States)

    Garro, Ana P; Chiapello, Laura S; Baronetti, José L; Masih, Diana T

    2011-01-01

    Experimental Cryptococcus neoformans infection in rats has been shown to have similarities with human cryptococcosis, revealing a strong granulomatous response and a low susceptibility to dissemination. Moreover, it has been shown that eosinophils are components of the inflammatory response to C. neoformans infections. In this in vitro study, we demonstrated that rat peritoneal eosinophils phagocytose opsonized live yeasts of C. neoformans, and that the phenomenon involves the engagement of FcγRII and CD18. Moreover, our results showed that the phagocytosis of opsonized C. neoformans triggers eosinophil activation, as indicated by (i) the up-regulation of major histocompatibility complex (MHC) class I, MHC class II and costimulatory molecules, and (ii) an increase in interleukin (IL)-12, tumour necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) production. However, nitric oxide (NO) and hydrogen peroxide (H2O2) synthesis by eosinophils was down-regulated after interaction with C. neoformans. Furthermore, this work demonstrated that CD4+ and CD8+ T lymphocytes isolated from spleens of infected rats and cultured with C. neoformans-pulsed eosinophils proliferate in an MHC class II- and class I-dependent manner, respectively, and produce important amounts of T-helper 1 (Th1) type cytokines, such as TNF-α and IFN-γ, in the absence of T-helper 2 (Th2) cytokine synthesis. In summary, the present study demonstrates that eosinophils act as fungal antigen-presenting cells and suggests that C. neoformans-loaded eosinophils might participate in the adaptive immune response. PMID:21039463

  20. Saccharomyces cerevisiae boulardii transient fungemia after intravenous self-inoculation

    OpenAIRE

    Cohen, Lola; Ranque, Stéphane; Raoult, Didier

    2013-01-01

    We report the case of a young psychotic intravenous drug user injecting herself with Saccharomyces cervisiae (boulardii). She experienced a 24 h fever, resolving spontaneously confirming, quasi experimentally, the inocuity of this yeast in a non-immunocompromised host.

  1. Susceptibility to heat and antifungal agents of Cryptococcus neoformans var. neoformans (serotype D isolated from Eucalyptus spp in Rio Grande do Sul, Brazil Susceptibilidade de Cryptococcus neoformans var. neoformans (sorotipo D isolados de Eucalyptus spp., no Rio Grande do Sul (Brasil, frente ao calor e a agentes antifúngicos

    Directory of Open Access Journals (Sweden)

    Jorge A Horta

    2005-03-01

    Full Text Available In this work we studied the susceptibility to heat and antifungal agents of the first strains of environmental Cryptococcus neoformans var neoformans (serotype D isolated in the state of Rio Grande do Sul, Brazil. In order to achieve a rigorous analysis, we employed the methodology recommended by NCCLS, Yeast Nitrogen Base (YNB proposed by Ghannoum et al (YNB-1, Antibiotic medium 3 (AM3 indicated by others, YNB adjusted to the NCCLS methodology (YNB-2 and Etest. Our results indicate that all strains were susceptible to amphotericin B (0.0625-0.5 µg/mL, fluconazole (0.125-8.0 µg/mL, itraconazole (0.031-0.25 µg/mL and flucytosine (0.125-4.0 µg/mL. The C. neoformans serotype D strains were more susceptible to heat (47ºC/30 min than C. neoformans serotype A.Este estudo foi realizado com os primeiros isolados ambientais de C. neoformans sorotipo D, obtidos no Rio Grande do Sul. Objetivando-se avaliar a susceptibilidade a agentes antifúngicos de forma mais rigorosa, utilizou-se a técnica de referência proposta pelo NCCLS, Caldo Yeast Nitrogen Base (YNB proposto por Ghannoum et al., Antibiotic medium 3, caldo YNB adequado à metodologia do NCCLS e o E-test. Os resultados indicaram que todos os isolados foram sensíveis à anfotericina B (0,0625-0,5 µg/mL, fluconazol (0,125-4,0 µg/mL, itraconazol (0,031-0,25 µg/ml e fluorocitosina (0,125-4,0 µg/mL através das técnicas empregadas. Nos testes de termotolerância (47ºC/30 min, observou-se que as culturas de C. neoformans sorotipo D são mais sensíveis do que as de C. neoformans sorotipo A.

  2. Cryptococcosis in captive cheetah (Acinonyx jubatus : two cases : case report

    Directory of Open Access Journals (Sweden)

    L.A. Bolton

    1999-07-01

    Full Text Available Cryptococcus neoformans is a yeast-like organism associated with pulmonary, meningoencephalitic, or systemic disease. This case report documents 2 cases of cryptococcosis with central nervous system involvement in captive cheetah (Acinonyx jubatus. In both cases the predominant post mortal lesions were pulmonary cryptococcomas and extensive meningoencephalomyelitis. Both cheetahs tested negative for feline immunodeficiency virus and feline leukaemia virus. The organism isolated in Case 2 was classified as Cryptococcus neoformans var. gattii, which is mainly associated with disease in immunocompetent hosts.

  3. Differential Antifungal Activity of Human and Cryptococcal Melanins with Structural Discrepancies

    OpenAIRE

    Néstor Correa; Néstor Correa; Néstor Correa; Cristian Covarrubias; Paula I. Rodas; Germán Hermosilla; Verónica R. Olate; Verónica R. Olate; Cristián Valdés; Wieland Meyer; Fabien Magne; Cecilia V. Tapia; Cecilia V. Tapia

    2017-01-01

    Melanin is a pigment found in all biological kingdoms, and plays a key role in protection against ultraviolet radiation, oxidizing agents, and ionizing radiation damage. Melanin exerts an antimicrobial activity against bacteria, fungi, and parasites. We demonstrated an antifungal activity of synthetic and human melanin against Candida sp. The members of the Cryptococcus neoformans and C. gattii species complexes are capsulated yeasts, which cause cryptococcosis. For both species melanin is an...

  4. The Case for Adopting the “Species Complex” Nomenclature for the Etiologic Agents of Cryptococcosis

    Directory of Open Access Journals (Sweden)

    Kyung J. Kwon-Chung

    2017-01-01

    Full Text Available Cryptococcosis is a potentially lethal disease of humans/animals caused by Cryptococcus neoformans and Cryptococcus gattii. Distinction between the two species is based on phenotypic and genotypic characteristics. Recently, it was proposed that C. neoformans be divided into two species and C. gattii into five species based on a phylogenetic analysis of 115 isolates. While this proposal adds to the knowledge about the genetic diversity and population structure of cryptococcosis agents, the published genotypes of 2,606 strains have already revealed more genetic diversity than is encompassed by seven species. Naming every clade as a separate species at this juncture will lead to continuing nomenclatural instability. In the absence of biological differences between clades and no consensus about how DNA sequence alone can delineate a species, we recommend using “Cryptococcus neoformans species complex” and “C. gattii species complex” as a practical intermediate step, rather than creating more species. This strategy recognizes genetic diversity without creating confusion.

  5. The status of cryptococcosis in Latin America

    Directory of Open Access Journals (Sweden)

    Carolina Firacative

    2018-04-01

    Full Text Available Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification of cryptococcal strains, in understanding their ecology, population genetics, host-pathogen interactions, and the clinical epidemiology of this important mycosis in Latin America.

  6. 新型隐球菌性脑膜炎与结核性脑膜炎的临床特点比较%Comparison of cli nical chara cteristics bet ween cryptococcus neoformans meningitis and tuberculous meningitis

    Institute of Scientific and Technical Information of China (English)

    李峤; 冯国栋; 高宇; 赵钢

    2016-01-01

    目的:对比和分析综合性医院新型隐球菌性脑膜炎(CNM)与结核性脑膜炎(TBM)的临床特点。方法分析62例 CNM 及219例 TBM 患者的一般资料、临床症状、脑脊液细胞学(CSFC)、头颅影像学的特点。结果两种脑膜炎均呈亚急性或慢性起病,相对于 TBM,CNM 延误诊断时间更长、临床症状中癫痫出现比率(24.2%)、头颅影像学中脑膜强化出现比率(17.7%)较高,腰穿脑脊液( CSF )压力(268.7±67.6mmH2 O)、CSF 单核细胞占比(21.0±17.5%)高,CSF 白细胞计数(84.8±88.1×106/L)、CSF 糖(1.9±1.4mmol・L-1)较低。结论两种脑膜炎临床表现不典型,两者之间不易鉴别,容易误诊漏诊。临床上需综合判断,CSF 瑞氏吉姆萨染色和改良抗酸染色有助于两者病原学确诊。%Objective To compare the clinical characteristics between cryptococcus neoformans meningitis and tuberculous meningitis in general hospitals .Methods Common date, clinical symptoms, cytology of cerebrospinal fluid, skull CT or MRI of 62 cases with cryptococcus neoformans meningitis and 219 cases with tuberculous meningitis were retrospectively analyzed .Results Both of these meningitis are sub-acute or chronic onset.Compared with tuberculous meningitis , duration of misdiagnosis is much longer in cryptococcus neoformans meningitis.There were significant differences in symptomatic epilepsy , cerebrospinal fluid pressure, proportion of the monocytes of CSF, white blood cell count of CSF ,glucose value of CSF, meningeal enhancement .Conclusion The clinical manifestations of these two meningitis are atypical .It is hard to identify and easily misdiagnosed .Careful observation and comprehensive judgments were required in the clinical work .Wright-Giemsa stain and modified acid-fast stain are conductive to etiological diagnosis .

  7. Colliding Epidemics and the Rise of Cryptococcosis

    Directory of Open Access Journals (Sweden)

    Christina C. Chang

    2015-12-01

    Full Text Available Discovered more than 100 years ago as a human pathogen, the Cryptococcus neoformans–Cryptococcus gattii (C. neoformans–C. gattii complex has seen a large global resurgence in its association with clinical disease in the last 30 years. First isolated in fermenting peach juice, and identified as a human pathogen in 1894 in a patient with bone lesions, this environmental pathogen has now found niches in soil, trees, birds, and domestic pets. Cryptococcosis is well recognized as an opportunistic infection and was first noted to be associated with reticuloendothelial cancers in the 1950s. Since then, advances in transplant immunology, medical science and surgical techniques have led to increasing numbers of solid organ transplantations (SOT and hematological stem cell transplantations being performed, and the use of biological immunotherapeutics in increasingly high-risk and older individuals, have contributed to the further rise in cryptococcosis. Globally, however, the major driver for revivification of cryptococcosis is undoubtedly the HIV epidemic, particularly in Sub-Saharan Africa where access to care and antiretroviral therapy remains limited and advanced immunodeficiency, poverty and malnutrition remains the norm. As a zoonotic disease, environmental outbreaks of both human and animal cryptococcosis have been reported, possibly driven by climate change. This is best exemplified by the resurgence of C. gattii infection in Vancouver Island, Canada, and the Pacific Northwest of the United States since 1999. Here we describe how the colliding epidemics of HIV, transplantation and immunologics, climate change and migration have contributed to the rise of cryptococcosis.

  8. Etude de quelques facteurs de pathogénicité liés aux infections fongiques hospitalières à Cryptococcus neoformans (production de biofilms et résistance

    OpenAIRE

    Mehiaoui, Samir

    2014-01-01

    ملخص عرفت السنوات الأخ رٌة انتشارا واسعا للأمراض الفطر ةٌ و الت أصبحت تشكل ف وقتنا الحاضر مضاعفات خط رٌة للمرضى ف المصالح الإستشفائ ةٌ. تعتبر الخم رٌة Cryptococcus neoformans واحدة من هذه الإصابات الجرثوم ةٌ القادرة على أن تسبب حالات من العدوى المكتسبة ف المستشف اٌت. مٌكن لهذه الخمائر أن تنمو ملامسة لبعض الأجهزة الطب ةٌ الحد ثٌة مثل القسطرات الور دٌ ةٌ الطرف ةٌ مشكلة الأغش ةٌ الح وٌ ةٌ و الت تتم زٌ بخاص ةٌ مقاومة للمضادات الفطر ةٌ. ف هذا الس اٌق، قمنا بإجراء دراسة عز ن لا من...

  9. Use of Pinus sylvestris L. (Pinaceae), Origanum vulgare L. (Lamiaceae), and Thymus vulgaris L. (Lamiaceae) essential oils and their main components to enhance itraconazole activity against azole susceptible/not-susceptible Cryptococcus neoformans strains.

    Science.gov (United States)

    Scalas, Daniela; Mandras, Narcisa; Roana, Janira; Tardugno, Roberta; Cuffini, Anna Maria; Ghisetti, Valeria; Benvenuti, Stefania; Tullio, Vivian

    2018-05-03

    Cryptococcal infections, besides being a problem for immunocompromised patients, are occasionally being a problem for immunocompetent patients. In addition, the lower susceptibility of this yeast to azoles is a growing problem in health care. To date, there are very few molecules with any activity towards Cryptococcus neoformans, leading to heightened interest in finding new alternatives or adjuvants to conventional drugs for the treatment of mycosis caused by this yeast. Since the essential oils (EOs) are considered as a potential rich source of bioactive antimicrobial compounds, we evaluated the antifungal activity of Origanum vulgare (oregano), Pinus sylvestris (pine), and Thymus vulgaris (thyme red) EOs, and their components (α-pinene, carvacrol, thymol) compared with fluconazole, itraconazole, and voriconazole, against C.neoformans clinical strains. Then, we investigated the effect of EOs and components in combination with itraconazole. EO composition was analysed by Gas chromatography-mass spectrometry (GC-MS). A broth microdilution method was used to evaluate the susceptibility of C.neoformans to azoles, EOs and components. Checkerboard tests, isobolograms and time-kill assays were carried out for combination studies. Six C.neoformans isolates were susceptible to azoles, while one C.neoformans exhibited a reduced susceptibility to all tested azole drugs. All EOs exerted a good inhibitory activity against all C.neoformans strains. Pine EO was the most effective. Among components, thymol exerted the most remarkable activity. By checkerboard testing and isobolographic analysis, combinations of itraconazole with oregano, pine, or thyme EOs, and carvacrol were found to be synergistic (FICI≤0.5) against azole susceptible C.neoformans. Regarding the azole not susceptible C.neoformans strain, the synergistic effect with itraconazole was observed with thyme EO (chemotype: thymol 26.52%; carvacrol 7.85%), and carvacrol. Time-kill assays confirmed the synergistic

  10. Implications of climate change (global warming) for the healthcare system.

    Science.gov (United States)

    Raffa, R B; Eltoukhy, N S; Raffa, K F

    2012-10-01

    Temperature-sensitive pathogenic species and their vectors and hosts are emerging in previously colder regions as a consequence of several factors, including global warming. As a result, an increasing number of people will be exposed to pathogens against which they have not previously needed defences. We illustrate this with a specific example of recent emergence of Cryptococcus gattii infections in more temperate climates. The outbreaks in more temperate climates of the highly virulent--but usually tropically restricted--C. gattii is illustrative of an anticipated growing challenge for the healthcare system. There is a need for preparedness by healthcare professionals in anticipation and for management of such outbreaks, including other infections whose recent increased prevalence in temperate climates can be at least partly associated with global warming. (Re)emergence of temperature-sensitive pathogenic species in more temperate climates will present new challenges for healthcare systems. Preparation for outbreaks should precede their occurrence. © 2012 Blackwell Publishing Ltd.

  11. Criptococoma pulmonar con invasión torácica en un varón inmunocompetente Pulmonary cryptococcoma with involvement of the chest wall in an immunocompetent patient

    Directory of Open Access Journals (Sweden)

    Ana A. Pisarevsky

    2010-04-01

    Full Text Available La criptococosis pulmonar en pacientes inmunocompetentes es una entidad poco habitual y su presentación como masas de gran tamaño, con compromiso de la pared torácica y de los tejidos blandos vecinos, no la encontramos descriptas en nuestra revisión bibliográfica. La variedad gattii (serotipoB/C está acotada geográficamente a regiones tropicales y subtropicales y parece afectar preferentemente a individuos inmunocompetentes. Presentamos el caso de un hombre de 51 años proveniente del noreste de la Argentina, fumador de 10 atados/año que consulta por aumento del volumen del hombro izquierdo e impotencia funcional por intenso dolor de seis meses de evolución. Mediante una biopsia percutánea de la masa, se diagnostica Cryptococcus neoformans, variedad gattii. El paciente recibe terapéutica antifúngica, mostrando una evolución favorable con disminución progresiva de la masa.The pulmonary cryptococcosis in immunocompetent patients is unusual, and its presentation as large masses with involvement of the chest wall and the neighboring soft tissues has not been found in our bibliographic research. The variety gattii (serotype B/C is limited geographically to tropical and subtropical regions and appears to affect particularly immunocompetent individuals. We describe the case of a 51-year-old man from the Northeast of Argentina, with a history of smoking 10 pack/year. He presented an increased volume of the left shoulder and reported that for the last six months he had been unable to move it due to the pain. A percutaneous biopsy of the mass provided a diagnosis of Cryptococcus neoformans, variety gattii. The patient was treated with antifungal therapy showing a favourable outcome with a progressive decrease of the mass.

  12. Differential Antifungal Activity of Human and Cryptococcal Melanins with Structural Discrepancies

    Directory of Open Access Journals (Sweden)

    Néstor Correa

    2017-07-01

    Full Text Available Melanin is a pigment found in all biological kingdoms, and plays a key role in protection against ultraviolet radiation, oxidizing agents, and ionizing radiation damage. Melanin exerts an antimicrobial activity against bacteria, fungi, and parasites. We demonstrated an antifungal activity of synthetic and human melanin against Candida sp. The members of the Cryptococcus neoformans and C. gattii species complexes are capsulated yeasts, which cause cryptococcosis. For both species melanin is an important virulence factor. To evaluate if cryptococcal and human melanins have antifungal activity against Cryptococcus species they both were assayed for their antifungal properties and physico-chemical characters. Melanin extracts from human hair and different strains of C. neoformans (n = 4 and C. gattii (n = 4 were investigated. The following minimum inhibitory concentrations were found for different melanins against C. neoformans and C. gattii were (average/range: 13.7/(7.8–15.6 and 19.5/(15.6–31.2 μg/mL, respectively, for human melanin; 273.4/(125–>500 and 367.2/(125.5–>500 μg/mL for C. neoformans melanin and 125/(62.5–250 and 156.2/(62–250 μg/mL for C. gattii melanin. Using Scanning Electron Microscopy we observed that human melanin showed a compact conformation and cryptococcal melanins exposed an amorphous conformation. Infrared spectroscopy (FTIR showed some differences in the signals related to C-C bonds of the aromatic ring of the melanin monomers. High Performance Liquid Chromatography established differences in the chromatograms of fungal melanins extracts in comparison with human and synthetic melanin, particularly in the retention time of the main compound of fungal melanin extracts and also in the presence of minor unknown compounds. On the other hand, MALDI-TOF-MS analysis showed slight differences in the spectra, specifically the presence of a minor intensity ion in synthetic and human melanin, as well as in some fungal

  13. Differential Antifungal Activity of Human and Cryptococcal Melanins with Structural Discrepancies.

    Science.gov (United States)

    Correa, Néstor; Covarrubias, Cristian; Rodas, Paula I; Hermosilla, Germán; Olate, Verónica R; Valdés, Cristián; Meyer, Wieland; Magne, Fabien; Tapia, Cecilia V

    2017-01-01

    Melanin is a pigment found in all biological kingdoms, and plays a key role in protection against ultraviolet radiation, oxidizing agents, and ionizing radiation damage. Melanin exerts an antimicrobial activity against bacteria, fungi, and parasites. We demonstrated an antifungal activity of synthetic and human melanin against Candida sp. The members of the Cryptococcus neoformans and C. gattii species complexes are capsulated yeasts, which cause cryptococcosis. For both species melanin is an important virulence factor. To evaluate if cryptococcal and human melanins have antifungal activity against Cryptococcus species they both were assayed for their antifungal properties and physico-chemical characters. Melanin extracts from human hair and different strains of C. neoformans ( n = 4) and C. gattii ( n = 4) were investigated. The following minimum inhibitory concentrations were found for different melanins against C. neoformans and C. gattii were (average/range): 13.7/(7.8-15.6) and 19.5/(15.6-31.2) μg/mL, respectively, for human melanin; 273.4/(125->500) and 367.2/(125.5->500) μg/mL for C. neoformans melanin and 125/(62.5-250) and 156.2/(62-250) μg/mL for C. gattii melanin. Using Scanning Electron Microscopy we observed that human melanin showed a compact conformation and cryptococcal melanins exposed an amorphous conformation. Infrared spectroscopy (FTIR) showed some differences in the signals related to C-C bonds of the aromatic ring of the melanin monomers. High Performance Liquid Chromatography established differences in the chromatograms of fungal melanins extracts in comparison with human and synthetic melanin, particularly in the retention time of the main compound of fungal melanin extracts and also in the presence of minor unknown compounds. On the other hand, MALDI-TOF-MS analysis showed slight differences in the spectra, specifically the presence of a minor intensity ion in synthetic and human melanin, as well as in some fungal melanin extracts. We

  14. ORIGINAL ARTICLES Cryptococcus and lymphocytic meningitis in ...

    African Journals Online (AJOL)

    2008-08-20

    Aug 20, 2008 ... of presumptive meningitis, we reviewed results of CSF cultures and cell .... AIDS patients with cryptococcal CSF infections.3 A third of the culture-positive cases ... is no indication that cryptococcal infection is unlikely. Thirdly,.

  15. presence of cryptococcus species in domestic chicken

    African Journals Online (AJOL)

    2009-05-25

    May 25, 2009 ... POSSIBLE RISK IT POSED TO HUMANS IN KABIGERIET VILLAGE, NAKURU COUNTY, KENYA. E.K. Kemoi, BSc, MSc, ... tree in the Brazilian Amazon rainforest (7). ... Sample collection: Environmental collection of domestic ...

  16. Comparison of BACTEC™ blood culture media for the detection of fungemia

    DEFF Research Database (Denmark)

    Datcu, Raluca; Boel, J; Jensen, I M

    2017-01-01

    with a positive blood culture with Candida species delivered to the Department of Clinical Microbiology, Herlev and Gentofte Hospital, Denmark in the 8-year period 2006 through 2014. The patients had at least one BACTEC™ aerobic and one Mycosis bottle sampled at the same time and at least one of the bottles...

  17. [Fungemia due to Trichosporon asahii in a patient with hematological malignancy].

    Science.gov (United States)

    Odero, Valle; Galán-Sánchez, Fátima; García-Agudo, Lidia; García-Tapia, Ana M; Guerrero-Lozano, Inmaculada; Rodríguez-Iglesias, Manuel A

    2015-01-01

    Trichosporonosis is an opportunistic infection caused by the genus Trichosporon. The majority of cases of invasive trichosporonosis occurs in immunocompromised individuals. We describe a case of disseminated infection by Trichosporon asahii in a hematology patient. A 52-year-old man diagnosed with acute lymphoblastic leukemia developed a febrile episode during the third cycle of the induction chemotherapy. The blood cultures were positive after 24h incubation, showing elongated structures compatible with fungal elements in the Gram stain. The identification of the fungus as Trichosporon asahii was carried out by the assimilation of compounds of carbon and the amplification and sequencing of the D1/D2 domain and the internal transcribed spacer of the ribosomal DNA. The fungus was also isolated from the pustular lesions that the patient had in the chest. After treatment with amphotericin B, the patient progressed satisfactorily. Trichosporon asahii is an emergent pathogen in immunosupressed patients and its presence should not be considered as colonization, as there is risk of invasive infection. Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  18. A case report describing detection of Rhodoturola minuta fungemia in an ewe lamb

    Science.gov (United States)

    An eight-month-old crossbred ewe that was normal upon physical examination was humanely euthanized for tissue collection. Prior to euthanasia, whole blood was collected via jugular venipuncture into 60-ml syringes containing EDTA anticoagulant. After sacrifice, the brain was removed and the choroi...

  19. Risk of Fungemia Due to Rhodotorula and Antifungal Susceptibility Testing of Rhodotorula Isolates

    OpenAIRE

    Zaas, Aimee K.; Boyce, Molly; Schell, Wiley; Lodge, Barbara Alexander; Miller, Jackie L.; Perfect, John R.

    2003-01-01

    Rhodotorula infections occur among patients with immunosuppression and/or central venous catheters. Using standardized methods (NCCLS M27-A), we determined the antifungal susceptibilities of 10 Rhodotorula bloodstream infection isolates. Patient information was collected for clinical correlation. The MICs of amphotericin B and posaconazole were the lowest, and the MICs of triazoles and echinocandins were higher than those of other antifungal agents.

  20. Risk of Fungemia Due to Rhodotorula and Antifungal Susceptibility Testing of Rhodotorula Isolates

    Science.gov (United States)

    Zaas, Aimee K.; Boyce, Molly; Schell, Wiley; Lodge, Barbara Alexander; Miller, Jackie L.; Perfect, John R.

    2003-01-01

    Rhodotorula infections occur among patients with immunosuppression and/or central venous catheters. Using standardized methods (NCCLS M27-A), we determined the antifungal susceptibilities of 10 Rhodotorula bloodstream infection isolates. Patient information was collected for clinical correlation. The MICs of amphotericin B and posaconazole were the lowest, and the MICs of triazoles and echinocandins were higher than those of other antifungal agents. PMID:14605170

  1. The analysis of protein differences in cerebrospinal fluid of tuberculous meningitis and cryptococcus meningitis by two-dimensional electrophoresis%应用双向电泳分析结核性脑膜炎与隐球菌性脑膜炎脑脊液的蛋白差异

    Institute of Scientific and Technical Information of China (English)

    黄绍梅; 杨瑜; 邱薇; 吴睿彦; 谢贝; 刘志辉; 陈志成

    2016-01-01

    Objective To screen the diagnostic markers between tuberculous meningitis and cryptococcus meningitis by analyzing the protein differences in cerebrospinal fluid (CSF).Methods The proteins in CSF of 4 cases of tuberculous meningitis and 4 cases of cryptococcal meningitis were separated by two-dimensional gel electrophoresis (2-DE).The differential proteins expression was descriptively analyzed according to the protein isoelectric point PI,molecular weight,matching rate,gray value by PDQuest8.0 software.Results 111 and 114 protein spots were captured from CSF of tuberculose meningitis and cryptococcus meningitis.The molecular weight of proteins ranged from 10 to 130 kDa.The isoelectric point (PI) was from 4.4 to 6.7.The matching rate of two groups was 95%.There were 24 differential proteins between two groups,of which 3 proteins were expressed only in tuberculous meningitis and 2 proteins expressed only in cryptococcal meningitis.19 matching protein point were two-fold differential expression.Conclusion 2-DE profiles with high resolution and reproducibility of CSF for tuberculous meningitis and cryptococcus meningitis are established,and some differences in protein expression exist between the two.%目的 比较分析结核性脑膜炎(TBM)与隐球菌性脑膜炎脑脊液的差异蛋白质,筛选二者的鉴别诊断标志物.方法 采用双向凝胶电泳技术对4例TBM和4例隐球菌性脑膜炎脑脊液蛋白进行电泳分离,以PDQuest8.0软件根据蛋白的等电点PI、分子量、匹配率、灰度值等进行差异蛋白描述性分析.结果 结核性、隐球菌性两组脑脊液分别检测到蛋白点数为111和114,蛋白点分子量均分布为10~130 kDa,等电点PI为4.4~6.7间.两组脑脊液蛋白图谱匹配率为95%,产生24个差异蛋白点,其中3个蛋白点只在TBM中表达,2个蛋白点只在隐球菌性脑膜炎中表达,19个匹配蛋白点存在2倍差异表达.结论 双向电泳技术可得到分辨率和重复率较高

  2. [Clinical problems in medical mycology: Problem number 51].

    Science.gov (United States)

    Romero, Mercedes; Messina, Fernando; Marín, Emmanuel; Arechavala, Alicia; Negroni, Ricardo; Depardo, Roxana; Walker, Laura; Benchetrit, Andrés; Santiso, Gabriela

    A 48 year-old immunocompetent woman, who had a nodular lesion in the neck and a dense infiltrate at the lower lobe of the left lung, presented at the Mycology Unit of Muñiz Hospital of Buenos Aires City. The pulmonary infiltrate disappeared spontaneously 3 months later. The histopathological study of the nodular lesion showed capsulated yeasts (mucicarmin and alcian blue positive stains) compatible with Cryptococcus. The mycological study of a new sample, obtained by a nodular puncture, allowed the isolation of yeasts, identified as Cryptococcus gattii (VGII). Latex test for Cryptococcus capsular antigen in serum was positive (1/100). CSF cultures rendered negative results. Fluconazole at a daily dose of 800mg was given during 45 days with partial improvement; as cultures from a new clinical sample were positive for Cryptococcus, the antimycotic was changed to itraconazole 400mg/day for 5 months, with an excellent clinical response. Copyright © 2017 Asociación Española de Micología. Publicado por Elsevier España, S.L.U. All rights reserved.

  3. Cryptococcosis outbreak in psittacine birds in Brazil.

    Science.gov (United States)

    Raso, T F; Werther, K; Miranda, E T; Mendes-Giannini, M J S

    2004-08-01

    An outbreak of cryptococcosis occurred in a breeding aviary in São Paulo, Brazil. Seven psittacine birds (of species Charmosyna papou, Lorius lory, Trichoglossus goldiei, Psittacula krameri and Psittacus erithacus) died of disseminated cryptococcosis. Incoordination, progressive paralysis and difficulty in flying were seen in five birds, whereas superficial lesions coincident with respiratory alterations were seen in two birds. Encapsulated yeasts suggestive of Cryptococcus sp. were seen in faecal smears stained with India ink in two cases. Histological examination of the birds showed cryptococcal cells in various tissues, including the beak, choana, sinus, lungs, air sacs, heart, liver, spleen, kidneys, intestines and central nervous system. High titres of cryptococcal antigen were observed in the serum of an affected bird. In this case, titres increased during treatment and the bird eventually died. Yeasts were isolated from the nasal mass, faeces and liver of one bird. Cryptococcus neoformans var. gattii serovar B was identified based on biochemical, physiological and serological tests. These strains were resistant (minimum inhibitory concentration 64 microg/ml) to fluconazole. This is the first report of C. neoformans var. gattii occurring in psittacine birds in Brazil.

  4. Fisetin as a promising antifungal agent against Cryptocococcus neoformans species complex.

    Science.gov (United States)

    Reis, M P C; Carvalho, C R C; Andrade, F A; Fernandes, O F L; Arruda, W; Silva, M R R

    2016-08-01

    The aim of this study was to investigate the mechanisms of action of fisetin, a flavonol with antifungal activity previously evaluated against the Cryptococcus neoformans species complex. Ergosterol content and flow cytometry analysis were determined for the C. neoformans species complex in the presence of fisetin and ultrastructural analysis of morphology was performed on Cryptococcus gattii and C. neoformans. Decrease in the total cellular ergosterol content after exposure to fisetin ranged from 25·4% after exposure to 128 μg ml(-1) to 21·6% after exposure to 64 μg ml(-1) of fisetin compared with the control (without fisetin). The fisetin effects obtained with flow cytometry showed metabolic impairment, and alterations in its normal morphology caused by fisetin in C. neoformans cells were verified using scanning electron microscopy. Fisetin is a compound that acts in the biosynthesis of ergosterol. Flow cytometry showed that fisetin reduced viability of the metabolically active cells of C. gattii, while morphological changes explain the action of fisetin in inhibiting growth of these fungi. This study supports the idea that fisetin may represent a good starting point for the development of future therapeutic substances for cryptococcosis. © 2016 The Society for Applied Microbiology.

  5. Bioactivity-guided isolation of laevicarpin, an antitrypanosomal and anticryptococcal lactam from Piper laevicarpu (Piperaceae).

    Science.gov (United States)

    da Silva A Maciel, Dayany; Freitas, Viviane P; Conserva, Geanne A Alves; Alexandre, Tatiana R; Purisco, Sonia U; Tempone, Andre G; Melhem, Márcia Souza C; Kato, Massuo J; Guimarães, Elsie F; Lago, João Henrique G

    2016-06-01

    Crude CH2Cl2 extract from leaves of Piper laevicarpu (Piperaceae) displayed antitrypanosomal activity against trypomastigote forms of Trypanosoma cruzi (Y strain) and antimicrobial potential against Cryptococcus gattii (strain-type WM 178). Bioactivity-guided fractionation of crude extract afforded one new natural bioactive lactam derivative, named laevicarpin. The structure of isolated compound, which displayed a very rare ring system, was elucidated based on NMR, IR and MS spectral analysis. Using MTT assay, the trypomastigotes of T. cruzi demonstrated susceptibility to laevicarpin displaying IC50 value of 14.7μg/mL (49.6μM), about 10-fold more potent than the standard drug benznidazole. The mammalian cytotoxicity of laevicarpin was verified against murine fibroblasts (NCTC cells) and demonstrated a CC50 value of 100.3μg/mL (337.7μM-SI=7). When tested against Cryptococcus gattii, laevicarpin showed an IC50 value of 2.3μg/mL (7.9μM) and a MIC value of 7.4μg/mL (25μM). Based in the obtained results, laevicarpin could be used as a scaffold for future drug design studies against the Chagas disease and anti-cryptococosis agents. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Cryptococcal infections over a 15 year period at a tertiary facility & impact of guideline management.

    Science.gov (United States)

    Gassiep, Ian; Douglas, Joel; Emeto, Theophilus I; Crawley, Katherine; Playford, Elliott G

    2018-04-17

    Cryptococcosis is an invasive fungal infection caused primarily by Cryptococcus neoformans and Cryptococcus gattii species, presenting predominantly as meningoencephalitis. The aim of this study is to assess all cryptococcal infections managed at our facility from 2001-2015 to determine incidence, risk factors, and comparison of outcomes prior to and following introduction of the 2010 Infectious Disease Society of America (IDSA) guidelines. Retrospective analysis of all patients diagnosed and treated for cryptococcal infection occurring between January 2001 and December 2015. Of 102 patients diagnosed with cryptococcal infection, 97 were eligible for study inclusion. There appears to be an overall increased incidence of cryptococcosis in both transplant and non-transplant cohorts with a peak in 2015 of 6 transplant and 13 non-transplant cases. In the meningitis cohort, 38/52 (73%) of identified isolates were C. neoformans, and 14/52 (27%) were C. gattii. Notably, 14/14 (100%) of C. gattii isolates were associated with meningitis, as compared to only 38/64 (59%) C. neoformans associated with meningitis (p: 0.003). It appears that patients presenting with cough are less likely to have meningitis, 17/27 (63%), (p: 0.005). When stratifying for culture positive meningitis lumbar puncture opening pressure, the median in the culture positive cohort was 31.5 cmH2O compared with 15.5 cmH2O (p: 0.036).Multiple admissions were required prior to diagnosis in the majority of cases with only 18/72 (25%) diagnosed on 1st presentation. Post-guideline mortality has improved from 15% to 6.1% (p: 0.046). Cryptococcal infection remains relatively uncommon, but there appears to be an increasing trend in incidence. Overall mortality is relatively low and has improved since introduction of the 2010 IDSA guidelines. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  7. The Effect of Novel Heterocyclic Compounds on Cryptococcal Biofilm

    Science.gov (United States)

    Korem, Maya; Kagan, Sarah

    2017-01-01

    Biofilm formation by microorganisms depends on their communication by quorum sensing, which is mediated by small diffusible signaling molecules that accumulate in the extracellular environment. During human infection, the pathogenic yeast Cryptococcus neoformans can form biofilm on medical devices, which protects the organism and increases its resistance to antifungal agents. The aim of this study was to test two novel heterocyclic compounds, S-8 (thiazolidinedione derivative, TZD) and NA-8 (succinimide derivative, SI), for their anti-biofilm activity against strains of Cryptococcus neoformans and Cryptococcus gattii. Biofilms were formed in a defined medium in 96-well polystyrene plates and 8-well micro-slides. The effect of sub-inhibitory concentrations of S-8 and NA-8 on biofilm formation was measured after 48 h by a metabolic reduction assay and by confocal laser microscopy analysis using fluorescent staining. The formation and development of cryptococcal biofilms was inhibited significantly by these compounds in concentrations below the minimum inhibitory concentration (MIC) values. These compounds may have a potential role in preventing fungal biofilm development on indwelling medical devices or even as a therapeutic measure after the establishment of biofilm. PMID:29371559

  8. Systemic fungal infections in patients with human inmunodeficiency virus.

    Science.gov (United States)

    Rodríguez-Cerdeira, C; Arenas, R; Moreno-Coutiño, G; Vásquez, E; Fernández, R; Chang, P

    2014-01-01

    Histoplasmosis is a systemic infection caused by the dimorphic fungus Histoplasma capsulatum. In immunocompromised patients, primary pulmonary infection can spread to the skin and meninges. Clinical manifestations appear in patients with a CD4(+) lymphocyte count of less than 150 cells/μL. Coccidioidomycosis is a systemic mycosis caused by Coccidioides immitis and Coccidioides posadasii. It can present as diffuse pulmonary disease or as a disseminated form primarily affecting the central nervous system, the bones, and the skin. Cryptococcosis is caused by Cryptococcus neoformans (var. neoformans and var. grubii) and Cryptococcus gattii, which are members of the Cryptococcus species complex and have 5 serotypes: A, B, C, D, and AD. It is a common opportunistic infection in patients with human immunodeficiency virus (HIV)/AIDS, even those receiving antiretroviral therapy. Histopathologic examination and culture of samples from any suspicious lesions are essential for the correct diagnosis of systemic fungal infections in patients with HIV/AIDS. Copyright © 2011 Elsevier España, S.L. and AEDV. All rights reserved.

  9. Análise proteômica de paracoccidioides sp. isolado de um caso de fungemia

    OpenAIRE

    Martins, Paulo Henrique Rosa

    2014-01-01

    Paracoccidioides spp. são agentes etiológicos da Paracoccidioidomicose uma micose sistêmica que afeta cerca de 10 milhões de pessoas nas regiões endêmicas. A incidência da doença é restrita à América Latina e a maioria dos casos são encontrados no Brasil. A doença é caracterizada por uma inflamação granulomatosa crônica, e os pacientes podem apresentar um amplo espectro de manifestações clínicas. Após a inalação de conídios, ocorre a instalação do fungo nos pulmões, que posteriormente através...

  10. Comparative evaluation of Oxoid Signal and BACTEC radiometric blood culture systems for the detection of bacteremia and fungemia

    International Nuclear Information System (INIS)

    Weinstein, M.P.; Mirrett, S.; Reller, L.B.

    1988-01-01

    The Oxoid Signal blood culture system is a newly described, innovative method for visually detecting growth of microorganisms. We did 5,999 paired comparisons of equal volumes (10 ml) of blood in the Oxoid Signal and BACTEC radiometric blood culture systems at two university hospitals that use identical methods of obtaining and processing specimens. Overall, more microorganisms were detected in the BACTEC system (P less than 0.001), in particular, streptococci (P less than 0.01), fungi (P less than 0.001), and nonfermentative gram-negative rods, especially Acinetobacter species (P less than 0.001). Trends favoring the BACTEC system for detection of Pseudomonas aeruginosa, Haemophilus species, and Neisseria species were noted. There were no differences in the yield of staphylococci, members of the family Enterobacteriaceae, and anaerobic bacteria. When both systems detected sepsis, the BACTEC did so earlier (P less than 0.001). This advantage was most notable at 24 h (70% of BACTEC positives detected versus 48% of Oxoid positives). The proportion of positives detected after 48 h, however, was similar (BACTEC, 84%; Oxoid, 78%). Revisions in the Oxoid Signal system itself or in the processing of Oxoid bottles appear to be necessary to improve its performance in detecting certain microorganism groups, especially fungi

  11. Brain and lung cryptococcoma and concurrent corynebacterium pseudotuberculosis infection in a goat: a case report

    Directory of Open Access Journals (Sweden)

    MCR Luvizotto

    2009-01-01

    Full Text Available A four-year-old male goat with a history of neurological disorder was euthanized. It presented uncommon nodules in the brain and lungs associated with multiple abscesses, predominantly in the spleen and liver. Histological examination of brain and lung sections revealed yeast forms confirmed to be Cryptococcus gattii after a combination of isolation and polymerase chain reaction (PCR procedures. Moreover, Corynebacterium pseudotuberculosis infection was diagnosed by PCR of samples from the lung, spleen and liver. The present report highlights the rare concurrent infection of C. gatti and C. pseudotuberculosis in an adult goat from São Paulo state, Brazil, and indicates the necessity of surveillance in the treatment of goats with atypical pulmonary infections associated with neurological disorders.

  12. Ebselen exerts antifungal activity by regulating glutathione (GSH) and reactive oxygen species (ROS) production in fungal cells.

    Science.gov (United States)

    Thangamani, Shankar; Eldesouky, Hassan E; Mohammad, Haroon; Pascuzzi, Pete E; Avramova, Larisa; Hazbun, Tony R; Seleem, Mohamed N

    2017-01-01

    Ebselen, an organoselenium compound and a clinically safe molecule has been reported to possess potent antifungal activity, but its antifungal mechanism of action and in vivo antifungal activity remain unclear. The antifungal effect of ebselen was tested against Candida albicans, C. glabrata, C. tropicalis, C. parapsilosis, Cryptococcus neoformans, and C. gattii clinical isolates. Chemogenomic profiling and biochemical assays were employed to identify the antifungal target of ebselen. Ebselen's antifungal activity in vivo was investigated in a Caenorhabditis elegans animal model. Ebselen exhibits potent antifungal activity against both Candida spp. and Cryptococcus spp., at concentrations ranging from 0.5 to 2μg/ml. Ebselen rapidly eradicates a high fungal inoculum within 2h of treatment. Investigation of the drug's antifungal mechanism of action indicates that ebselen depletes intracellular glutathione (GSH) levels, leading to increased production of reactive oxygen species (ROS), and thereby disturbs the redox homeostasis in fungal cells. Examination of ebselen's in vivo antifungal activity in two Caenorhabditis elegans models of infection demonstrate that ebselen is superior to conventional antifungal drugs (fluconazole, flucytosine and amphotericin) in reducing Candida and Cryptococcus fungal load. Ebselen possesses potent antifungal activity against clinically relevant isolates of both Candida and Cryptococcus by regulating GSH and ROS production. The potent in vivo antifungal activity of ebselen supports further investigation for repurposing it for use as an antifungal agent. The present study shows that ebselen targets glutathione and also support that glutathione as a potential target for antifungal drug development. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. The antimicrobial activity of lapachol and its thiosemicarbazone and semicarbazone derivatives

    Directory of Open Access Journals (Sweden)

    Marina Azevedo Souza

    2013-05-01

    Full Text Available Lapachol was chemically modified to obtain its thiosemicarbazone and semicarbazone derivatives. These compounds were tested for antimicrobial activity against several bacteria and fungi by the broth microdilution method. The thiosemicarbazone and semicarbazone derivatives of lapachol exhibited antimicrobial activity against the bacteria Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentrations (MICs of 0.05 and 0.10 µmol/mL, respectively. The thiosemicarbazone and semicarbazone derivatives were also active against the pathogenic yeast Cryptococcus gattii (MICs of 0.10 and 0.20 µmol/mL, respectively. In addition, the lapachol thiosemicarbazone derivative was active against 11 clinical isolates of Paracoccidioides brasiliensis, with MICs ranging from 0.01-0.10 µmol/mL. The lapachol-derived thiosemicarbazone was not cytotoxic to normal cells at the concentrations that were active against fungi and bacteria. We synthesised, for the first time, thiosemicarbazone and semicarbazone derivatives of lapachol. The MICs for the lapachol-derived thiosemicarbazone against S. aureus, E. faecalis, C. gattii and several isolates of P. brasiliensis indicated that this compound has the potential to be developed into novel drugs to treat infections caused these microbes.

  14. Clinical and microbiological features of cryptococcal meningitis

    Directory of Open Access Journals (Sweden)

    Lucia Kioko Hasimoto e Souza

    2013-06-01

    Full Text Available Introduction In this study, the clinical features, underlying diseases and clinical outcomes of patients with cryptococcosis were investigated. In addition, a molecular analysis of the Cryptococcus neoformans species complex isolated from these patients was performed. Methods A prospective study of 62 cases of patients with cryptococcal infection was conducted at the Hospital de Doenças Tropicais de Goiás Dr. Anuar Auad from 2009-2010. Cryptococcal meningitis cases were diagnosed by direct examination and cerebrospinal fluid (CSF sample culture. The profiling of these patients was assessed. The CSF samples were submitted to India ink preparation and cultured on Sabouraud dextrose agar, and C. neoformans was identified by the production of urease, a positive phenoloxidase test and assimilation of carbohydrates. C. neoformans and C. gattii isolates were distinguished by growth on L-canavanine-glycine-bromothymol blue medium, and molecular analysis was conducted via PCR fingerprinting reactions using M13 and (GACA4 primers. Results From the 62 patients with cryptococcosis, 71 isolates of CSF were obtained; 67 (94.4% isolates were identified as C. neoformans var. grubii/VNI, and 4 (5.6% were identified as C. gattii/VGII. Of these patients, 53 had an HIV diagnosis. The incidence of cryptococcosis was higher among patients 20-40 years of age, with 74.2% of the cases reported in males. Cryptococcus-related mortality was noted in 48.4% of the patients, and the symptoms were altered sensorium, headache, fever and stiff neck. Conclusions The high morbidity and mortality observed among patients with cryptococcosis demonstrate the importance of obtaining information regarding the epidemiological profile and clinical course of the disease in the State of Goiás, Brazil.

  15. [Cryptococcus neoformans meningitis in children and adolescents infected with HIV].

    Science.gov (United States)

    Quian, Jorge; Gutiérrez, Stella; González, Virginia; Sánchez, Mercedes; Abayian, Marina; Baccino, Fernando

    2012-10-01

    Three cases of meningitis due to criptococcus in children infected with HIV are reported. All patients had severe immunodeficiency. The first case occurred before antiretroviral therapy. The second case had poor adherence to treatment. The third case is a teenager with prolonged fever, who did not have a clear source of transmission of HIV and was immunodeficient at the time of diagnosis. Examination of cerebrospinal fluid with India ink dye was essential for making the diagnosis in all three cases. The disease course was adequate in all three patients despite flucytosine not being available.

  16. Amino Acid Permeases and Virulence in Cryptococcus neoformans.

    Directory of Open Access Journals (Sweden)

    Kevin Felipe Cruz Martho

    Full Text Available Fungal opportunistic pathogens colonize various environments, from plants and wood to human and animal tissue. Regarding human pathogens, one great challenge during contrasting niche occupation is the adaptation to different conditions, such as temperature, osmolarity, salinity, pressure, oxidative stress and nutritional availability, which may constitute sources of stress that need to be tolerated and overcome. As an opportunistic pathogen, C. neoformans faces exactly these situations during the transition from the environment to the human host, encountering nutritional constraints. Our previous and current research on amino acid biosynthetic pathways indicates that amino acid permeases are regulated by the presence of the amino acids, nitrogen and temperature. Saccharomyces cerevisiae and Candida albicans have twenty-four and twenty-seven genes encoding amino acid permeases, respectively; conversely, they are scarce in number in Basidiomycetes (C. neoformans, Coprinopsis cinerea and Ustilago maydis, where nine to ten permease genes can be found depending on the species. In this study, we have demonstrated that two amino acid permeases are essential for virulence in C. neoformans. Our data showed that C. neoformans uses two global and redundant amino acid permeases, Aap4 and Aap5 to respond correctly to thermal and oxidative stress. Double deletion of these permeases causes growth arrest in C. neoformans at 37°C and in the presence of hydrogen peroxide. The inability to uptake amino acid at a higher temperature and under oxidative stress also led to virulence attenuation in vivo. Our data showed that thermosensitivity caused by the lack of permeases Aap4 and Aap5 can be remedied by alkaline conditions (higher pH and salinity. Permeases Aap4 and Aap5 are also required during fluconazole stress and they are the target of the plant secondary metabolite eugenol, a potent antifungal inhibitor that targets amino acid permeases. In summary, our work unravels (i interesting physiological property of C. neoformans regarding its amino acid uptake system; (ii an important aspect of virulence, which is the need for amino acid permeases during thermal and oxidative stress resistance and, hence, host invasion and colonization; and (iii provides a convenient prototype for antifungal development, which are the amino acid permeases Aap4/Aap5 and their inhibitor.

  17. 21 CFR 866.3165 - Cryptococcus neoformans serological reagents.

    Science.gov (United States)

    2010-04-01

    ... SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3165... clinical specimens or from cultured isolates derived from clinical specimens. The identification aids in...

  18. cryptococcus meningitis in a cohort of hiv positive kenyan patients

    African Journals Online (AJOL)

    the acquired immunodeficiency syndrome. N. Engl. J. Med. 1997; 337:15-21. 7. Khanna N., Chandramuki A., Desai A., et al. Cryptococcol infections of the central nervous system: An analysis of predisposing factors, laboratory findings and outcome in patients from South India with special reference to HIV infection. J. Med.

  19. In silico approaches in the identification of Cryptococcus ...

    African Journals Online (AJOL)

    taylorsadmin

    2012-06-07

    Jun 7, 2012 ... alignment was performed using the Clustal W software. exigua and Spodoptera litura based on the knowledge of highly conserved chemoreceptors in Drosophila melanogaster. Their research group employed bioinformatics approaches to identify the desired sequences and web-based TMHMM software to.

  20. Development and validation of an extended database for yeast identification by MALDI-TOF MS in Argentina.

    Science.gov (United States)

    Taverna, Constanza Giselle; Mazza, Mariana; Bueno, Nadia Soledad; Alvarez, Christian; Amigot, Susana; Andreani, Mariana; Azula, Natalia; Barrios, Rubén; Fernández, Norma; Fox, Barbara; Guelfand, Liliana; Maldonado, Ivana; Murisengo, Omar Alejandro; Relloso, Silvia; Vivot, Matias; Davel, Graciela

    2018-05-11

    Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) has revolutionized the identification of microorganisms in clinical laboratories because it is rapid, relatively simple to use, accurate, and can be used for a wide number of microorganisms. Several studies have demonstrated the utility of this technique in the identification of yeasts; however, its performance is usually improved by the extension of the database. Here we developed an in-house database of 143 strains belonging to 42 yeast species in the MALDI Biotyper platform, and we validated the extended database with 388 regional strains and 15 reference strains belonging to 55 yeast species. We also performed an intra- and interlaboratory study to assess reproducibility and analyzed the use of the cutoff values of 1.700 and 2.000 to correctly identify at species level. The creation of an in-house database that extended the manufacturer's database was successful in view of no incorrect identification was introduced. The best performance was observed by using the extended database and a cutoff value of 1.700 with a sensitivity of .94 and specificity of .96. A reproducibility study showed utility to detect deviations and could be used for external quality control. The extended database was able to differentiate closely related species and it has potential in distinguishing the molecular genotypes of Cryptococcus neoformans and Cryptococcus gattii.

  1. Broad-Range PCR Coupled with Electrospray Ionization Time of Flight Mass Spectrometry for Detection of Bacteremia and Fungemia in Patients with Neutropenic Fever

    Science.gov (United States)

    Maertens, J.; Bueselinck, K.; Lagrou, K.

    2016-01-01

    Infection is an important complication in patients with hematologic malignancies or solid tumors undergoing intensive cytotoxic chemotherapy. In only 20 to 30% of the febrile neutropenic episodes, an infectious agent is detected by conventional cultures. In this prospective study, the performance of broad-range PCR coupled with electrospray ionization time of flight mass spectrometry (PCR/ESI-MS) technology was compared to conventional blood cultures (BC) in a consecutive series of samples from high-risk hematology patients. In 74 patients, BC and a whole-blood sample for PCR/ESI-MS (Iridica BAC BSI; Abbott, Carlsbad, CA, USA) were collected at the start of each febrile neutropenic episode and, in case of persistent fever, also at day 5. During 100 different febrile episodes, 105 blood samples were collected and analyzed by PCR/ESI-MS. There was evidence of a bloodstream infection (BSI) in 36/105 cases (34%), based on 14 cases with both PCR/ESI-MS and BC positivity, 17 cases with BC positivity only, and 5 cases with PCR/ESI-MS positivity only. The sensitivity of PCR/ESI-MS was 45%, specificity was 93%, and the negative predictive value was 80% compared to blood culture. PCR/ESI-MS detected definite pathogens (Fusobacterium nucleatum and Streptococcus pneumoniae) missed by BC, whereas it missed both Gram-negative and Gram-positive organisms detected by BC. PCR/ESI-MS testing detected additional microorganisms but showed a low sensitivity (45%) compared to BC in neutropenic patients. Our results indicate a lower concordance between BC and PCR/ESI-MS in the neutropenic population than what has been previously reported in other patient groups with normal white blood cell distribution, and a lower sensitivity than other PCR-based methods. PMID:27440820

  2. Deteksi Antigen pada Kriptokokosis

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    Robiatul  Adawiyah

    2014-12-01

    Full Text Available AbstrakKriptokokosis  merupakan  infeksi  sistemik  yang  disebabkan  Cryptococcus  sp.  Predileksi jamur tersebut adalah susunan saraf pusat dan selaput otak. Terdapat 5 spesies Cryptococcus sp. yang menyebabkan penyakit pada manusia; yang paling banyak adalah Cr. neoformans dan Cr.  gattii.  Diagnosis kriptokokosis ditegakkan berdasarkan gejala klinis, pemeriksaan laboratoris PCR. Pemeriksaan secara morfologi dengan tinta India positif  bila jumlah sel jamur 10 sel/ml spesimen. Kultur dilakukan di media sabouraud dextrose agar (SDA dan niger sheed agar (NSA, jamur  tumbuh  setelah  5­7  hari.  Deteksi  antigen  dan  antibodi  dilakukan  pada  cairan  tubuh  dan tidak membutuhkan waktu lama. Deteksi antibodi Cr.neoformans memiliki kelemahan yaitu tidak menunjukkan hasil positif pada infeksi akut, IgA masih positif setelah 1­2 tahun fase penyembuhan, IgG  dapat  persisten,  pada individu  imunokompromis menunjukkan hasil  yang  sangat  kompleks dan dalam menentukan diagnosis sering tidak  konsisten. Polisakarida  adalah komponen paling berperan dalam virulensi Cr. neoformans. Komponen polisakarida terutama glucuronoxylomannan merupakan petanda penting dalam diagnosis kriptokokosis secara serologis. Deteksi antigen Cr. neoformans memiliki kelebihan yaitu menunjukkan hasil positif pada infeksi akut/kronis, sensitivitas antigen yang minimal tetap dapat mendiagnosis kriptokokosis. Kata kunci: Cr. neoformans, glucuronoxylomannan, antigenAbstractCryptococcosis is systemic infection that caused by Cryptococcus sp. Predilection of this fungi is the central nervous system and brain membrane. There are 5 species of Cryptococcus sp. that cause  cryptococcosis  in  human;  but  the  majority  are  caused  by  Cr.  neoformans  and  Cr.  gattii. The diagnosis of cryptococcosis is made based on clinical symptoms, laboratory and radiological PCR. Morphological examination with India ink is positive when the

  3. Deteksi Antigen pada Kriptokokosis

    Directory of Open Access Journals (Sweden)

    Robiatul Adawiyah

    2014-12-01

    Full Text Available AbstrakKriptokokosis merupakan infeksi sistemik yang disebabkan Cryptococcus sp. Predileksi jamur tersebut adalah susunan saraf pusat dan selaput otak. Terdapat 5 spesies Cryptococcus sp. yang menyebabkan penyakit pada manusia; yang paling banyak adalah Cr. neoformans dan Cr. gattii. Diagnosis kriptokokosis ditegakkan berdasarkan gejala klinis, pemeriksaan laboratoris serta radiologis. Pemeriksaan laboratoris dilakukan dengan identifikasi morfologi, serologi danPCR. Pemeriksaan secara morfologi dengan tinta India positif  bila jumlah sel jamur 10  sel/ml spesimen. Kultur dilakukan di media sabouraud dextrose agar (SDA dan niger sheed agar (NSA, jamur tumbuh setelah 5-7 hari. Deteksi antigen dan antibodi dilakukan pada cairan tubuh dan tidak membutuhkan waktu lama. Deteksi antibodi Cr.neoformans memiliki kelemahan yaitu tidak menunjukkan hasil positif pada infeksi akut, IgA masih positif setelah 1-2 tahun fase penyembuhan, IgG dapat persisten, pada individu imunokompromis menunjukkan hasil yang sangat kompleks dan dalam menentukan diagnosis sering tidak konsisten. Polisakarida adalah komponen paling berperan dalam virulensi Cr. neoformans. Komponen polisakarida terutama glucuronoxylomannan merupakan petanda penting dalam diagnosis kriptokokosis secara serologis. Deteksi antigen Cr. neoformans memiliki kelebihan yaitu menunjukkan hasil positif pada infeksi akut/kronis, sensitivitas dan spesifisitas tinggi, dapat mendeteksi polisakarida hingga 10 ng/ml sehingga dengan kadarantigen yang minimal tetap dapat mendiagnosis kriptokokosis.Kata kunci: Cr. neoformans, glucuronoxylomannan, antigenAbstractCryptococcosis is systemic infection that caused by Cryptococcus sp. Predilection of this fungi is the central nervous system and brain membrane. There are 5 species of Cryptococcus sp. that cause cryptococcosis in human; but the majority are caused by Cr. neoformans and Cr. gattii. The diagnosis of cryptococcosis is made based on clinical symptoms

  4. Evidence of Cryptococcosis in cattle in Zaria Kaduna state, Nigeria

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    Emmanuella N. Akange

    2013-04-01

    Full Text Available Aim: Cryptococcosis is azoonotic infection caused by fungal of the Cryptococcus neoformans complex comprising of C. neoformans and C. gattii.The disease affects humans and animals worldwide causing morbidity and mortality. This work was carried out to determine the occurrence of cryptococcal antigens and factors associated with presence of antigens in cattle in Zaria, Nigeria. Materials and Methods: Three hundred and ninety (390 serum samples from cattle of various ages were collected from 11 farms in Zaria, Nigeria. The samples were analysed using alatex agglutination test and lateral flow assay kit which detectsthe polysaccharide capsular antigens of Cryptococcus species. Results:Out of the 390 samples tested 28 (7.17% were found to be positive using the latex agglutination test while only of these 22 (5.64% were positive using the lateral flow assay. There was a strong correlation (r=0.939, p=0.0002 between the results of the latex agglutination test and the lateral flow assay. There was no statistically significant difference (p>0.005 in positivity for cryptococcal antigens between sex, age and sex, though, there was a statistically significant difference (p<0.05 in positivity between management systems i.e. semi-intensive and intensive farming systems. Conclusions: The epidemiological value of this report lies in its demonstration that the risk of cattle and humans infection with cryptococcosis exist in farms in Zaria. The presence of this pathogen among these cattle poses an economic threat to the livestock industry due to the mastitis it causes. It also poses a significant public health threat because of its zoonotic nature and the increasing population of immunocompromised individuals. Large scale studies to determine specific risk factors and the role of the environment and experimental studies to determine what governs the transition from nasal colonisation to infection are recommended. [Vet World 2013; 6(2.000: 64-67

  5. Identification of cultured isolates of clinically important yeast species using fluorescent fragment length analysis of the amplified internally transcribed rRNA spacer 2 region

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    Muylaert An

    2002-07-01

    Full Text Available Abstract Background The number of patients with yeast infection has increased during the last years. Also the variety of species of clinical importance has increased. Correct species identification is often important for efficient therapy, but is currently mostly based on phenotypic features and is sometimes time-consuming and depends largely on the expertise of technicians. Therefore, we evaluated the feasibility of PCR-based amplification of the internally transcribed spacer region 2 (ITS2, followed by fragment size analysis on the ABI Prism 310 for the identification of clinically important yeasts. Results A rapid DNA-extraction method, based on simple boiling-freezing was introduced. Of the 26 species tested, 22 could be identified unambiguously by scoring the length of the ITS2-region. No distinction could be made between the species Trichosporon asteroides and T. inkin or between T. mucoides and T. ovoides. The two varieties of Cryptococcus neoformans (var. neoformans and var. gattii could be differentiated from each other due to a one bp length difference of the ITS2 fragment. The three Cryptococcus laurentii isolates were split into two groups according to their ITS2-fragment lengths, in correspondence with the phylogenetic groups described previously. Since the obtained fragment lengths compare well to those described previously and could be exchanged between two laboratories, an internationally usable library of ITS2 fragment lengths can be constructed. Conclusions The existing ITS2 size based library enables identification of most of the clinically important yeast species within 6 hours starting from a single colony and can be easily updated when new species are described. Data can be exchanged between laboratories.

  6. Cryptococcal cerebellitis in no-VIH patient.

    Science.gov (United States)

    Lasso, Fabricio Andres; Zamora Bastidas, Tomas Omar; Potosí García, Jorge Andrés; Díaz Idrobo, Bairon

    2017-06-30

    Cryptococcosis is an opportunistic fungal infection whose etiology is Cryptococcus neofromans / C. gattii, complex which affects immunocompromised patients mainly. Meningeal infection is one of the most common presentations, but cerebellar affection is rare. Male patient with 65 old years, from an area of subtropical climate with chronic exposure to poultry, without pathological antecedents, who presented clinical picture consistent with headache, fever, seizures and altered mental status. Initially without menigeal signs or intracranial hypertension and normal neurological examination. Later, the patient developed ataxia, dysdiadochokinesia and limb loss. By lumbar punction and image of nuclear magnetic resonance (NMR) cerebellitis cryptococcal was diagnosticated. Antifungal therapy with amphotericin B and fluconazole was performed, however the patient died. The cryptococcosis has different presentations, it´s a disease whose incidence has been increasing since the advent of the HIV / AIDS pandemy, however the commitment of the encephalic parenchyma and in particular the cerebellum is considered rare. In this way we are facing the first case of cryptococcal cerebellitis in our midst.

  7. Sexual Reproduction of Human Fungal Pathogens

    Science.gov (United States)

    Heitman, Joseph; Carter, Dee A.; Dyer, Paul S.; Soll, David R.

    2014-01-01

    We review here recent advances in our understanding of sexual reproduction in fungal pathogens that commonly infect humans, including Candida albicans, Cryptococcus neoformans/gattii, and Aspergillus fumigatus. Where appropriate or relevant, we introduce findings on other species associated with human infections. In particular, we focus on rapid advances involving genetic, genomic, and population genetic approaches that have reshaped our view of how fungal pathogens evolve. Rather than being asexual, mitotic, and largely clonal, as was thought to be prevalent as recently as a decade ago, we now appreciate that the vast majority of pathogenic fungi have retained extant sexual, or parasexual, cycles. In some examples, sexual and parasexual unions of pathogenic fungi involve closely related individuals, generating diversity in the population but with more restricted recombination than expected from fertile, sexual, outcrossing and recombining populations. In other cases, species and isolates participate in global outcrossing populations with the capacity for considerable levels of gene flow. These findings illustrate general principles of eukaryotic pathogen emergence with relevance for other fungi, parasitic eukaryotic pathogens, and both unicellular and multicellular eukaryotic organisms. PMID:25085958

  8. Cryptococcosis in the koala (Phascolarctos cinereus): pathogenesis and treatment in the context of two atypical cases.

    Science.gov (United States)

    Schmertmann, Laura J; Stalder, Kathryn; Hudson, Donald; Martin, Patricia; Makara, Mariano; Meyer, Wieland; Malik, Richard; Krockenberger, Mark B

    2018-02-24

    Disseminated cryptococcosis caused by Cryptococcus gattii (molecular type VGI) was diagnosed in an adult free-ranging female koala (Phascolarctos cinereus). Subclinical cryptococcosis was later diagnosed in this koala's joey. In the adult koala, a pathological fracture of the tibia was associated with the bone lysis of marked focal cryptococcal osteomyelitis. Limb-sparing orthopedic intervention, in the setting of disseminated cryptococcosis, was judged to have a poor prognosis, and the adult koala was euthanized. The joey was removed and hand-reared. Serological testing revealed persistent and increasing cryptococcal capsular antigenemia in the absence of clinical signs of disease and it was subsequently treated with oral fluconazole for approximately 16 months, rehabilitated and released into the wild. It was sighted 3 months post-release in a good state of health and again at 18 months post-release but was not recaptured on either occasion. This is the first published report of cryptococcal appendicular osteomyelitis in a koala. It is also the first report of concurrent disease in a dependent juvenile and the successful treatment of subclinical cryptococcosis to full resolution of the cryptococcal antigenemia in a free-ranging koala. This paper provides a discussion of cryptococcal osteomyelitis in animals, host-pathogen-environment interactions and treatment and monitoring protocols for cryptococcosis in koalas. Published reports describing the treatment of cryptococcosis in koalas are also collated and summarised.

  9. Aislamiento de cryptococcus neoformans var. Grubii en un felino doméstico. Reporte de caso

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    Marlyn Zapata Montoya

    2009-12-01

    Full Text Available Se halló evidencia clínica, citológica y serológica de una infección natural con Criptococcus en un felino criollo proveniente de La Mesa, Cundinamarca (Colombia. C. neoformans var. grubii fue aislado del cultivo de secreciones nasales del paciente. Para el tratamiento específico se utilizó fluconazol, que mostró buenos resultados a partir del primer mes de haber iniciado su administración.

  10. Aislamiento de cryptococcus neoformans var. Grubii en un felino doméstico. Reporte de caso

    OpenAIRE

    Zapata Montoya, Marlyn; Galindo Zamora, Vladimir

    2009-01-01

    Se halló evidencia clínica, citológica y serológica de una infección natural con Criptococcus en un felino criollo proveniente de La Mesa, Cundinamarca (Colombia). C. neoformans var. grubii fue aislado del cultivo de secreciones nasales del paciente. Para el tratamiento específico se utilizó fluconazol, que mostró buenos resultados a partir del primer mes de haber iniciado su administración. Clinical, cytologic and serologic evidence of a natural infection with Criptococcus was identified ...

  11. Aislamiento de cryptococcus neoformans var. Crubii en un felino doméstico. Reporte de caso

    OpenAIRE

    Zapata Montoya, Marlyn; Galindo Zamora, Vladimir

    2009-01-01

    Se halló evidencia clínica, citológica y serológica de una infección natural con Criptococcus en un felino criollo proveniente de La Mesa, Cundinamarca (Colombia). C. neoformans var. grubii fue aislado del cultivo de secreciones nasales del paciente. Para el tratamiento específico se utilizó fluconazol, que mostró buenos resultados a partir del primer mes de haber iniciado su administración. Clinical, cytologic and serologic evidence of a natural infection with Criptococcus was identified ...

  12. Combined therapy of interleukin-12 and interleukin-18 against cryptococcus neoformans infection in a murine model

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Objective To explore adverse effects of combined treatment of interleukin-12 (IL-12) and interleukin-18 (IL-18) against cryptococcosis in a murine model.Methods Infected mice were treated with a combination of IL-12 and IL-18. Their body weight and intake of water and food were observed and recorded. Serum levels of leptin were detected with an enzyme-linked immuno sorbent assay (ELISA).Results In the combined treatment group, the intake volume of water and food were reduced, leading to weight loss and undetectable levels of leptin in the serum. These adverse effects were more profound in mice that had received higher doses of cytokines, which sometimes led to a fatal outcome. There was a significant difference compared with the control group. Neutralization of endogenous tumor necrosis factor-α (TNF-α) by its specific mAb did not alter the wasting effect of this treatment.Conclusions The combined IL-12/IL-18 treatment may cause a number of adverse effects independent of TNF-α and leptin synthesis. Further investigations for resolving these adverse effects are required before clinical application of these cytokines.

  13. Dectin-3 Is Not Required for Protection against Cryptococcus neoformans Infection.

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    Althea Campuzano

    Full Text Available C-type lectin receptors (CLRs are diverse, trans-membrane proteins that function as pattern recognition receptors (PRRs which are necessary for orchestrating immune responses against pathogens. CLRs have been shown to play a major role in recognition and protection against fungal pathogens. Dectin-3 (also known as MCL, Clecsf8, or Clec4d is a myeloid cell-specific CLR that recognizes mycobacterial trehalose 6,6'-dimycolate (TDM as well as α-mannans present in the cell wall of fungal pathogens. To date, a potential role for Dectin-3 in the mediation of protective immune responses against C. neoformans has yet to be determined. Consequently, we evaluated the impact of Dectin-3 deficiency on the development of protective immune responses against C. neoformans using an experimental murine model of pulmonary cryptococcosis. Dectin-3 deficiency did not lead to increased susceptibility of mice to experimental pulmonary C. neoformans infection. Also, no significant differences in pulmonary leukocyte recruitment and cytokine production were observed in Dectin-3 deficient mice compared to wild type infected mice. In addition, we observed no differences in uptake and anti-cryptococcal activity of Dectin-3 deficient dendritic cells and macrophages. Altogether, our studies show that Dectin-3 is dispensable for mediating protective immune responses against pulmonary C. neoformans infection.

  14. Allelic exchange of pheromones and their receptors reprograms sexual identity in Cryptococcus neoformans.

    OpenAIRE

    Brynne C Stanton; Steven S Giles; Mark W Staudt; Emilia K Kruzel; Christina M Hull

    2010-01-01

    Cell type specification is a fundamental process that all cells must carry out to ensure appropriate behaviors in response to environmental stimuli. In fungi, cell identity is critical for defining "sexes" known as mating types and is controlled by components of mating type (MAT) loci. MAT-encoded genes function to define sexes via two distinct paradigms: 1) by controlling transcription of components common to both sexes, or 2) by expressing specially encoded factors (pheromones and their rec...

  15. Isolation and molecular characterization of Cryptococcus species isolated from pigeon nests and Eucalyptus trees

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    Abuzar Kamari

    2017-06-01

    Conclusion: The presence of pigeons and Eucalyptus trees in the vicinity of some particular places such as rest homes and hospitals should be considered as a risk factor for the immunocompromised population.  

  16. Naïve B cells reduce fungal dissemination in Cryptococcus neoformans infected Rag1-/- mice.

    Science.gov (United States)

    Dufaud, Chad; Rivera, Johanna; Rohatgi, Soma; Pirofski, Liise-Anne

    2018-01-01

    IgM and B-1 cell deficient mice exhibit early C. neoformans dissemination from lungs to brain, but a definitive role for B cells in conferring resistance to C. neoformans dissemination has not been established. To address this question, we developed an intranasal (i.n.) C. neoformans infection model in B and T cell deficient Rag1 -/- mice and found they also exhibit earlier fungal dissemination and higher brain CFU than wild-type C57Bl/6 (wild-type) mice. To probe the effect of B cells on fungal dissemination, Rag1 -/- mice were given splenic (intravenously) or peritoneal (intraperitoneally) B cells from wild-type mice and infected i.n. with C. neoformans 7 d later. Mice that received B cells had lung histopathology resembling wild type mice 14 d post-infection, and B-1, not B-2 or T cells in their lungs, and serum and lung IgM and IgG 21 d post-infection. Lung CFU were comparable in wild-type, Rag1 -/-, and Rag1 -/- mice that received B cells 21 d post-infection, but brain CFU were significantly lower in mice that received B cells than Rag1 -/- mice that did not. To determine if natural antibody can promote immunity in our model, we measured alveolar macrophage phagocytosis of C. neoformans in Rag1 -/- mice treated with naive wild-type IgM-sufficient or sIgM -/- IgM-deficient sera before infection. Compared to IgM-deficient sera, IgM-sufficient sera significantly increased phagocytosis. Our data establish B cells are able to reduce early C. neoformans dissemination in mice and suggest natural IgM may be a key mediator of early antifungal immunity in the lungs.

  17. Fatal Saccharomyces Cerevisiae Aortic Graft Infection

    Science.gov (United States)

    Meyer, Michael (Technical Monitor); Smith, Davey; Metzgar, David; Wills, Christopher; Fierer, Joshua

    2002-01-01

    Saccharomyces cerevisiae is a yeast commonly used in baking and a frequent colonizer of human mucosal surfaces. It is considered relatively nonpathogenic in immunocompetent adults. We present a case of S. cerevisiae fungemia and aortic graft infection in an immunocompetent adult. This is the first reported case of S. cerevisiue fungemia where the identity of the pathogen was confirmed by rRNA sequencing.

  18. Multidrug-resistant endemic clonal strain of Candida auris in India

    NARCIS (Netherlands)

    Chowdhary, A.; Kumar, V.; Sharma, C.; Prakash, A.; Agarwal, K.; Babu, R.; Dinesh, K.R.; Karim, S.; Singh, S.K.; Hagen, F.; Meis, J.F.G.M.

    2014-01-01

    Candida auris is a recently described rare agent of fungemia. It is notable for its antifungal resistance. A total of 15 C. auris isolates, originating from seven cases of fungemia, three cases of diabetic gangrenous foot, and one case of bronchopneumonia from a tertiary care hospital in south

  19. Pharmacokinetics of fluconazole following intravenous and oral administration to koalas (Phascolarctos cinereus).

    Science.gov (United States)

    Black, L A; Krockenberger, M B; Kimble, B; Govendir, M

    2014-02-01

    Clinically normal koalas (n = 12) received a single dose of 10 mg/kg fluconazole orally (p.o.; n = 6) or intravenously (i.v.; n = 6). Serial plasma samples were collected over 24 h, and fluconazole concentrations were determined using a validated HPLC assay. A noncompartmental pharmacokinetic analysis was performed. Following i.v. administration, median (range) plasma clearance (CL) and steady-state volume of distribution (Vss ) were 0.31 (0.11-0.55) L/h/kg and 0.92 (0.38-1.40) L/kg, respectively. The elimination half-life (t1/2 ) was much shorter than in many species (i.v.: median 2.25, range 0.98-6.51 h; p.o.: 4.69, range 2.47-8.01 h), and oral bioavailability was low and variable (median 0.53, range 0.20-0.97). Absorption rate-limited disposition was evident. Plasma protein binding was 39.5 ± 3.5%. Although fluconazole volume of distribution (Varea ) displayed an allometric relationship with other mammals, CL and t1/2 did not. Allometrically scaled values were approximately sevenfold lower (CL) and sixfold higher (t1/2 ) than observed values, highlighting flaws associated with this technique in physiologically distinct species. On the basis of fAUC/MIC pharmacodynamic targets, fluconazole is predicted to be ineffective against Cryptococcus gattii in the koala as a sole therapeutic agent administered at 10 mg/kg p.o. every 12 h. © 2013 John Wiley & Sons Ltd.

  20. [Antimycotic activity in vitro and in vivo of 5-fluorocytosine on pathogenic strains of Candida albicans and Cryptococcus neoformans].

    Science.gov (United States)

    Costa, A L; Valenti, A; Costa, G; Calogero, F

    1976-01-01

    The authors have analyzed the 5 Fluoro Cytosine (5FC) activity on strains of Candida albicans and Criptococcus neoformans, both in vitro and in vivo. In vitro the minimal inhibitory concentration (MIC) was determined; in vivo tests of pathogenicity on rabbit and mouse have been executed. The various findings obtained have shown a strong activity of the 5FC on strains of Candida and Criptococcus.

  1. Determinación de anticuerpos contra Cryptococcus neoformans en un grupo de colombófilos

    OpenAIRE

    Carmen Sofía Corrales; Nelly Ordóñez; Luisa Marcela Londoño; Elizabeth Castañeda

    1987-01-01

    Se evaluó la respuesta humoral, medida como anticuerpos específicos anti-Criptococcus neoformans, en 69 sueros de un grupo de colombófilos de Bogotá; grupo considerado como de alto riesgo dado el frecuente contacto con el material contaminado con el microorganismo. Además se estudiaron 65 sueros de un grupo control constituído por personas dedicadas a actividades diferentes. En las pruebas de inmunofluorescencia indirecta y aglutinación en tubo, no se evidenció diferencia entre los niveles de...

  2. Paradoxical immune reconstitution inflammatory syndrome associated with previous Cryptococcus neoformans infection in an HIV-positive patient requiring neurosurgical intervention.

    Science.gov (United States)

    Biagetti, Carlo; Nicola, Monica; Borderi, Marco; Pavoni, Michele; Tampellini, Livia; Verucchi, Gabriella; Chiodo, Francesco

    2009-04-01

    Immune reconstitution inflammatory syndrome (IRIS) in HIV-1-infected patients is associated with an exaggerated inflammatory response against an opportunistic infection during highly active antiretroviral therapy. The only review on IRIS associated with Criptococcus neoformans reported 21 episodes including lymphadenitis, necrotizing pneumonitis, breast and cutaneous abscess, and cryptococcomas. To our knowledge this is the first report of IRIS associated with previous meningeal criptococcal infection which required neurosurgical intervention with placement of a ventriculo-peritoneal shunt to drain a CSF cyst formed by exclusion of the temporal horn of the right lateral ventricle. We demonstrate that this procedure is possible without complications such as cryptococcal dissemination into the peritoneum.

  3. Determinación de anticuerpos contra Cryptococcus neoformans en un grupo de colombófilos

    Directory of Open Access Journals (Sweden)

    Carmen Sofía Corrales

    1987-12-01

    Full Text Available Se evaluó la respuesta humoral, medida como anticuerpos específicos anti-Criptococcus neoformans, en 69 sueros de un grupo de colombófilos de Bogotá; grupo considerado como de alto riesgo dado el frecuente contacto con el material contaminado con el microorganismo. Además se estudiaron 65 sueros de un grupo control constituído por personas dedicadas a actividades diferentes. En las pruebas de inmunofluorescencia indirecta y aglutinación en tubo, no se evidenció diferencia entre los niveles de anticuerpos de ambos grupos. Se estudiaron adicionalmente 15 palomares con el objeto de aislar el microorganismo lo que se logró en el 66,7% de ellos. En 8 de los aislamientos se hizo serotipificación del hongo encontrándose que la totalidad correspondía a C. neoformans var. neoformans.

  4. Genome-wide expression profiling of the response to short-term exposure to fluconazole in Cryptococcus neoformans serotype A

    Directory of Open Access Journals (Sweden)

    Sanguinetti Maurizio

    2011-05-01

    Full Text Available Abstract Background Fluconazole (FLC, a triazole antifungal drug, is widely used for the maintenance therapy of cryptococcal meningoencephalitis, the most common opportunistic infection in AIDS patients. In this study, we examined changes in the gene expression profile of the C. neoformans reference strain H99 (serotype A following FLC treatment in order to investigate the adaptive cellular responses to drug stress. Results Simultaneous analysis of over 6823 transcripts revealed that 476 genes were responsive to FLC. As expected up-regulation of genes involved in ergosterol biosynthesis was observed, including the azole target gene ERG11 and ERG13, ERG1, ERG7, ERG25, ERG2, ERG3 and ERG5. In addition, SRE1 which is a gene encoding a well-known regulator of sterol homeostasis in C. neoformans was up-regulated. Several other genes such as those involved in a variety of important cellular processes (i.e. lipid and fatty acid metabolism, cell wall maintenance, stress and virulence were found to be up-regulated in response to FLC treatment. Conversely, expression of AFR1, the major transporter of azoles in C. neoformans, was not regulated by FLC. Conclusions Short-term exposure of C. neoformans to FLC resulted in a complex altered gene expression profile. Some of the observed changes could represent specific adaptive responses to the antifungal agent in this pathogenic yeast.

  5. Genomic analysis of Bacillus subtilis OH 131.1 and coculturing with Cryptococcus flavescens for control of fusarium head blight

    Science.gov (United States)

    Bacillus subtilis OH131.1 is a bacterial antagonist of Fusarium graminearum, a plant pathogen which causes Fusarium head blight in wheat. The genome of B. subtilis OH131.1 was sequenced, annotated and analyzed to understand its potential to produce bioactive metabolites. The analysis identified 6 sy...

  6. Low diversity Cryptococcus neoformans variety grubii multilocus sequence types from Thailand are consistent with an ancestral African origin

    NARCIS (Netherlands)

    Simwami, S.P.; Khayhan, K.; Henk, D.A.; Aanensen, D.M.; Boekhout, T.; Hagen, F.; Brouwer, A.E.; Harrison, T.S.; Donnelly, C.A.; Fisher, M.D.

    2011-01-01

    The global burden of HIV-associated cryptococcal meningitis is estimated at nearly one million cases per year, causing up to a third of all AIDS-related deaths. Molecular epidemiology constitutes the main methodology for understanding the factors underpinning the emergence of this understudied, yet

  7. Heavy metal tolerance in the psychrotolerant Cryptococcus sp. isolated from deep-sea sediments of the Central Indian Basin

    Digital Repository Service at National Institute of Oceanography (India)

    Singh, P.; Raghukumar, C.; Parvatkar, R.R.; Mascarenhas-Pereira, M.B.L.

    of the heavy metal salts i.e., ZnSO sub(4) , CuSO sub(4), Pb(CH sub(3) COO) sub(2) and CdCl sub(2). It demonstrated considerable growth in the presence of 100 mg/l concentrations of the above-mentioned four heavy metal salts both at 30 degrees C and 15 degrees...

  8. Differentially expressed genes under simulated deep-sea conditions in the psychrotolerant yeast Cryptococcus sp. NIOCCPY13

    Digital Repository Service at National Institute of Oceanography (India)

    Singh, P.; Raghukumar, C.; Verma, A.K.; Meena, R.M.

    Epoxide hydrolase [Rhodotorula mucilaginosa], AAV64029.1 Arachidonic acid metabolism 58 2e-22 PH_13 Hypothetical Protein RTG_00533 [Rhodotorula glutinis ATCC 204091], EGU13356.1 Amp binding enzyme 85 2e-29 PH_15 C-4 methylsterol oxidase [Puccinia..., PTH_30 Hypothetical protein SNOG_09178 [Phaeosphaeria nodorum SN15], XP_001799479.1 AMP-activated protein kinase 52 2.7 PTH_25 Family 2 glycosyltransferase [Melampsora larici-populina 98AG31], EGF99634.1 Catalyzes the incorporation of GlcNAc from...

  9. Polysaccharides and phenolic compounds as substrate for yeasts isolated from rotten wood and description of Cryptococcus fagi sp.nov.

    NARCIS (Netherlands)

    Middelhoven, W.J.

    2006-01-01

    Pieces of rotten wood collected in the forest were screened for the presence of yeasts. In spring time 3 tree species were sampled, followed by 9 species in summer. Yeast strains were identified by traditional methods. Identifications were confirmed by sequencing of ribosomal DNA in case of doubt.

  10. Anti-cryptococcal activity of ethanol crude extract and hexane fraction from Ocimum basilicum var. Maria bonita: mechanisms of action and synergism with amphotericin B and Ocimum basilicum essential oil.

    Science.gov (United States)

    Cardoso, Nathalia N R; Alviano, Celuta S; Blank, Arie F; Arrigoni-Blank, Maria de Fátima; Romanos, Maria Teresa V; Cunha, Marcel M L; da Silva, Antonio Jorge R; Alviano, Daniela S

    2017-12-01

    Ocimum basilicum L. (Lamiaceae) has been used in folk medicine to treat headaches, kidney disorders, and intestinal worms. This study evaluates the anti-cryptococcal activity of ethanol crude extract and hexane fraction obtained from O. basilicum var. Maria Bonita leaves. The MIC values for Cryptococcus sp. were obtained according to Clinical and Laboratory Standards Institute in a range of 0.3-2500 μg/mL. The checkerboard assay evaluated the association of the substances tested (in a range of 0.099-2500 μg/mL) with amphotericin B and O. basilicum essential oil for 48 h. The ethanol extract, hexane fraction and associations in a range of 0.3-2500 μg/mL were tested for pigmentation inhibition after 7 days of treatment. The inhibition of ergosterol synthesis and reduction of capsule size were evaluated after the treatment with ethanol extract (312 μg/mL), hexane fraction (78 μg/mL) and the combinations of essential oil + ethanol extract (78 μg/mL + 19.5 μg/mL, respectively) and essential oil + hexane fraction (39.36 μg/mL + 10 μg/mL, respectively) for 24 and 48 h, respectively. The hexane fraction presented better results than the ethanol extract, with a low MIC (156 μg/mL against C. neoformans T 444 and 312 μg/mL against C. neoformans H99 serotype A and C. gattii WM779 serotype C). The combination of the ethanol extract and hexane fraction with amphotericin B and essential oil enhanced their antifungal activity, reducing the concentration of each substance needed to kill 100% of the inoculum. The substances tested were able to reduce the pigmentation, capsule size and ergosterol synthesis, which suggest they have important mechanisms of action. These results provide further support for the use of ethanol extracts of O. basilicum as a potential source of antifungal agents.

  11. Clinical utility of the cryptococcal antigen lateral flow assay in a diagnostic mycology laboratory.

    Directory of Open Access Journals (Sweden)

    Brendan J McMullan

    Full Text Available BACKGROUND: Cryptococcus neoformans causes life-threatening meningitis. A recently introduced lateral flow immunoassay (LFA to detect cryptococcal antigen (CRAG is reportedly more rapid and convenient than standard latex agglutination (LA, but has not yet been evaluated in a diagnostic laboratory setting. METHODS: One hundred and six serum, 42 cerebrospinal fluid (CSF, and 20 urine samples from 92 patients with known or suspected cryptococcosis were tested by LA and LFA, and titres were compared. Results were correlated with laboratory-confirmed cryptococcosis. Serial samples were tested in nine treated patients. RESULTS: Twenty-five of 92 patients had confirmed cryptococcosis; all sera (n = 56 from these patients were positive by LFA (sensitivity 100%, 95% confidence interval (CI 93.6-100% compared with 51/56 positive by LA (sensitivity 91.1%, 95% CI 80.7-96.1%. Fifty sera from 67 patients without cryptococcosis tested negative in both assays. While LA yielded more false negative results (5/56 this did not reach statistical significance (p = 0.063. Nine CSF samples from patients with cryptococcal meningitis yielded positive results using both assays while 17/18 urine samples from patients with cryptococcosis were positive by the LFA. The LFA detected CRAG in C. gattii infection (n = 4 patients. Agreement between titres obtained by both methods (n = 38 samples was imperfect; correlation between log-transformed titres (r was 0.84. Turn-around-time was 20 minutes for the LFA and 2 h for LA. The cost per qualitative sample was 18USD and 91 USD, respectively and per quantitative sample was 38USD and 144USD, respectively. CONCLUSIONS: Qualitative agreement between the LFA and LA assays performed on serum and CSF was good but agreement between titres was imperfect. Ease of performance of the LFA and the capacity for testing urine suggest it has a role in the routine laboratory as a rapid diagnostic test or point-of-care test.

  12. ORF Alignment: NC_003074 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available in CNBN0180 [Cryptococcus neoformans var. ... neoformans B-3501A] gb|AAW47011.1| nucleolus protein, ... ... ... putative [Cryptococcus neoformans var. neoformans JEC21] ... ref|XP_568528.1| nucleolus pro

  13. Infecção por Cryptococcus limitada à próstata em paciente aidético com micobacteriose disseminada. Relato de necropsia

    OpenAIRE

    Lima, Marcus Aurelho de; Santos, Jenner A. Modesto dos; Lazo, Javier; Silva-Vergara, Mario León; Santos, Lister A. Modesto dos; Santos, Vitorino Modesto dos

    1997-01-01

    Relata-se caso de infecção criptocócica confinada à próstata, como achado de necropsia, em homem de 32 anos portador da síndrome da imunodeficiência adquirida (SIDA) com micobacteriose disseminada. Enfatiza-se a importância do achado incidental em necropsia e a persistência dessa infecção micótica na próstata.This is a case report of asymptomatic and restrained cryptococcal prostatic infection in a 32-year old black male with the acquired immunodeficiency syndrome, whose death was caused by s...

  14. Integrated Activity and Genetic Profiling of Secreted Peptidases in Cryptococcus neoformans Reveals an Aspartyl Peptidase Required for Low pH Survival and Virulence

    Czech Academy of Sciences Publication Activity Database

    Clarke, S. C.; Dumesic, P. A.; Homer, C. M.; O'Donoghue, A.J.; La Greca, F.; Pallová, Lenka; Majer, Pavel; Madhani, H. D.; Craik, C. S.

    2016-01-01

    Roč. 12, č. 12 (2016), č. článku e1006051. ISSN 1553-7366 Institutional support: RVO:61388963 Keywords : protease inhibitors * Candida albicans * parasitic protozoa Subject RIV: CC - Organic Chemistry Impact factor: 6.608, year: 2016 http://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1006051&type=printable

  15. Impact of process parameters on the sourdough microbiota, selection of suitable starter strains, and description of the novel yeast Cryptococcus thermophilus sp. nov.

    OpenAIRE

    Vogelmann, Stephanie Anke

    2013-01-01

    The microbiota of a ripe sourdough consists of lactic acid bacteria (LAB), especially of the genus Lactobacillus, and yeasts. Their composition is influenced by the interplay of species or strains, the kind of substrate as well as the process parameters temperature, dough yield, redox potential, refreshment time, and number of propagation steps (Hammes and Gänzle, 1997). As taste and quality of sourdough breads are mainly influenced by the fermentation microbiota, intense research has been fo...

  16. Immune-Mediated Damage Completes the Parabola: Cryptococcus neoformans Pathogenesis Can Reflect the Outcome of a Weak or Strong Immune Response

    Directory of Open Access Journals (Sweden)

    Liise-anne Pirofski

    2017-12-01

    Full Text Available Cryptococcosis occurs most frequently in immunocompromised individuals. This has led to the prevailing view that this disease is the result of weak immune responses that cannot control the fungus. However, increasingly, clinical and experimental studies have revealed that the host immune response can contribute to cryptococcal pathogenesis, including the recent study of L. M. Neal et al. (mBio 8:e01415-17, 2017, https://doi.org/10.1128/mBio.01415-17 that reports that CD4+ T cells mediate tissue damage in experimental murine cryptococcosis. This finding has fundamental implications for our understanding of the pathogenesis of cryptococcal disease; it helps explain why immunotherapy has been largely unsuccessful in treatment and provides insight into the paradoxical observation that HIV-associated cryptococcosis may have a better prognosis than cryptococcosis in those with no known immune impairment. The demonstration that host-mediated damage can drive cryptococcal disease provides proof of concept that the parabola put forth in the damage-response framework has the flexibility to depict complex and changing outcomes of host-microbe interaction.

  17. Cryptococcosis

    Science.gov (United States)

    C. neoformans var. neoformans infection; C. neoformans var. gatti infection; C. neoformans var. grubii infection ... C. neoformans and C. gattii are the fungi that cause this disease. Infection with C. neoformans is ...

  18. Clinical, microbiological, and experimental animal studies of Candida lipolytica.

    Science.gov (United States)

    Walsh, T J; Salkin, I F; Dixon, D M; Hurd, N J

    1989-01-01

    Candida lipolytica was recovered from six patients in three different clinical centers. The index isolate caused a persistent fungemia with catheter-associated Candida thrombophlebitis, the second isolate was from a polymicrobial sinusitis, and the remaining four isolates were involved in tissue colonization. These and 20 other isolates were consistent in their morphological and physiological characteristics. All formed true hyphae and blastoconidia on cornmeal-Tween 80 agar and all assimilated glucose, glycerol, and erythritol. In a murine model of disseminated candidiasis, the index isolate that caused clinical fungemia caused no mortality and produced only two lesions on a kidney, as determined at necropsy. The nine isolates selected for in vitro antifungal susceptibility studies had intermediate susceptibilities to amphotericin B but were susceptible to ketoconazole. We conclude that C. lipolytica is a weakly virulent pathogen which may require an intravascular foreign body to cause fungemia. Images PMID:2745702

  19. Bloodstream infections by Malassezia and Candida species in critical care patients.

    NARCIS (Netherlands)

    Iatta, R.; Cafarchia, C.; Cuna, T.; Montagna, M.T.; Laforgia, N.; Gentile, O.; Rizzo, A.; Boekhout, T.; Otranto, D.

    2013-01-01

    Despite being considered an emerging yeast related to immunocompromised individuals, severe infections by Malassezia furfur have not been evaluated. During a one-year survey on yeasts fungemia, 290 neonatal and 17 pediatric patients with intravascular catheters, lipid parenteral nutrition, prolonged

  20. Uncommon opportunistic yeast bloodstream infections from Qatar

    NARCIS (Netherlands)

    Taj-Aldeen, S.J.; AbdulWahab, A.; Kolecka, A.; Deshmukh, A.; Meis, J.F.G.M.; Boekhout, T.

    2014-01-01

    Eleven uncommon yeast species that are associated with high mortality rates irrespective of antifungal therapy were isolated from 17/187 (201 episodes) pediatric and elderly patients with fungemia from Qatar. The samples were taken over a 6-year period (January 2004-December 2010). Isolated species

  1. Molecular Tools for Cryptic "Candida" Species Identification with Applications in a Clinical Laboratory

    Science.gov (United States)

    Gamarra, Soledad; Dudiuk, Catiana; Mancilla, Estefania; Vera Garate, Maria Veronica; Guerrero, Sergio; Garcia-Effron, Guillermo

    2013-01-01

    "Candida" spp. includes more than 160 species but only 20 species pose clinical problems. "C. albicans" and "C. parapsilosis" account for more than 75% of all the fungemias worldwide. In 1995 and 2005, one "C. albicans" and two "C. parapsilosis"-related species were described, respectively. Using…

  2. Lack of IL-6 increases blood–brain barrier permeability in fungal

    Indian Academy of Sciences (India)

    Interleukin (IL-6) is a multifunctional cytokine, and numerous studies have shown that IL‐6 influences the integrity of the blood–brain barrier. In this study we investigated the role of IL-6 in Cryptococcus meningitis. First, wild-type or IL-6−/− mice were injected with Cryptococcus neoformans (C. neoformans) and the survival ...

  3. Rapid Methods for the Laboratory Identification of Pathogenic Microorganisms.

    Science.gov (United States)

    1982-09-01

    coli Hemophilus influenzae Bacillus anthracis Bacillus circulans Bacillus coagulans Bacillus cereus T Candida albicans Cryptococcus neoformans Legionel...reveree aide If neceeeary and Identify by block number) Lectins: Rapid Identification, Bacillus anthracisjCryptococcus " neoformans. Neisseria...field-type kit for the rapid identification of Bacillus anthracis. We have shown that certain lectins will selectively interact with B. anthracis

  4. LOOP mediated isothermal AMPlification (LAMP) in diagnosis of ...

    African Journals Online (AJOL)

    We collected a quantity of 35 serum samples of HIVpositive patients and a number of 107 cerebrospinal fluid (CSF) samples of patients who had shown symptoms of meningitis. We designed target specific primers for PCR and LAMP techniques to trace C. neoformans and C. gattii. From the total 142 clinical specimens, five ...

  5. Candida dubliniensis spondylodiscitis in an immunocompetent patient. Case report and review of the literature

    OpenAIRE

    Oksi, Jarmo; Finnilä, Taru; Hohenthal, Ulla; Rantakokko-Jalava, Kaisu

    2014-01-01

    We describe what appears to be the first case of spondylodiscitis due to Candida dubliniensis. Our case adds to the current literature of the importance of C. dubliniensis as a cause of fungemia and subsequent deep seated infections. It highlights the importance of taking fungal as well as bacterial culture from biopsy specimens in patients with spondylodiscitis. We also review the literature covering the reported cases of Candida spondylodiscitis, which amount to about 100 over the last 5 de...

  6. Voriconazole in an infant with cryptococcal meningitis

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    @@ Cryptococcus neoformans (C. neoformans) is the most common cause of fungal meningitis worldwide.1 Cryptococcal meningitis is an opportunistic infection commonly found in immunocompromised hosts,especially HIV-infected adults. It also occurs in apparently immunocompetent individuals.

  7. LOOP mediated isothermal AMPlification (LAMP) in diagnosis of ...

    African Journals Online (AJOL)

    2012-02-28

    Feb 28, 2012 ... Due to disadvantages of known ... advantages, as compared to traditional diagnostic methods like ... Products of LAMP reaction are DNA fragments with stemloop ..... "Differentiation of Cryptococcus neoformans varieties and.

  8. CHARACTERIZATION ADN BIOLOGICAL ACTIVITY OF SECONDARY METABOLITES FROM ARMILLARIA TABESCENS

    Science.gov (United States)

    Ethyl acetate extracts from liquid cultures of Armillaria tabescens showed good antimicrobial activity against Candida albicans, Cryptococcus neoformans, Escherichia coli and Mycobacterium intracellulare. Chemical analyses of extract constituents led to the isolation and identification of two new co...

  9. Screening of antibacterial and antifungal activities in green and ...

    African Journals Online (AJOL)

    ... bacteria: Bacillus cereus, Bacillus thuringiensis, Bacillus subtillus, Clostridium sporogenes, Staphylococcus aureus, Mycobacterium smegmatis, Streptococcus faecalis and Bacillus sp, two Gram-negative bacteria: Escherichia coli and Pseudomonas sp and against fungi: Candida tropicalis and Cryptococcus neoformans.

  10. Cryptococcal capsular glucuronoxylomannan reduces ischaemia-related neutrophil influx

    NARCIS (Netherlands)

    Ellerbroek, PM; Schoemaker, RG; van Veghel, R; Hoepelman, AIM; Coenjaerts, FEJ

    Background The capsular polysaccharide glucuronoxylomannan (GXM) of Cryptococcus neoformans interferes with the chemotaxis and transendothelial migration of neutrophils. Intravenous administration of purified GXM has been shown to reduce the influx of inflammatory cells in an animal model of

  11. In vitro anidulafungin activity against yeasts – system and disseminated mycosis pathogens

    Directory of Open Access Journals (Sweden)

    A. B. Kulko

    2015-01-01

    Full Text Available We analyzed susceptibility to anidulafungin of yeasts clinical strains of Candida (14 species, Cryptococcus (1 species, Geotrichum (1 species, Rhodotorula (1 species and Saccharomyces (1 species. We revealed high anidulafungin activity against Candida spp., both common species and rare pathogens of candidiasis. It was found that over 99 % of Candida strains do not have an acquired resistance mechanisms to anidulafungin (microbiological criteria. The anidulafungin is not active against strains of Cryptococcus neoformans and Rhodotorula mucilaginosa.

  12. Sucessfull management of bilateral presumed Candida endogenous endophtalmitis following pancreatitis

    Directory of Open Access Journals (Sweden)

    Ricardo Evangelista Marrocos de Aragão

    2016-06-01

    Full Text Available ABSTRACT Endogenous endophthalmitis is a rare, and frequently devastating, ophthalmic disease. It occurs mostly in immunocompromised patients, or those with diabetes mellitus, cancer or intravenous drugs users. Candida infection is the most common cause of endogenous endophthalmitis. Ocular candidiasis develops within days to weeks of fungemia. The association of treatment for pancreatitis with endophthalmitis is unusual. Treatment with broad-spectrum antibiotics and total parenteral nutrition may explain endogenous endophthalmitis. We report the case of a patient with pancreatitis treated with broad-spectrum antibiotics and total parenteral nutrition who developed bilateral presumed Candida endogenous endophthalmitis that was successfully treated with vitrectomy and intravitreal amphotericin B.

  13. Isolated pulmonary candidiasis in a patient with diabetes mellitus: A rare case report.

    Science.gov (United States)

    Hakamifard, Atousa; Khorvash, Farzin; Raisi, Arash

    2016-01-01

    Fungal infections are as a cause of morbidity and mortality in immunocompromise patients. Because the respiratory tract is colonized with Candida, the presence of this agent in respiratory specimens makes the diagnosis of Candida pneumonia problematic. Candida pneumonia is a rare infection, and the majority of cases are secondary to hematogenous dissemination. Furthermore isolated Candida pneumonia originating from endotracheal inoculation is an extremely rare entity. We describe a case of isolated pulmonary candidiasis in the form multiple nodular lesions in a patient with long-term history of diabetes mellitus without evidence of fungemia or systemic involvement who responded to antifungal therapy.

  14. Isolated pulmonary candidiasis in a patient with diabetes mellitus: A rare case report

    Directory of Open Access Journals (Sweden)

    Atousa Hakamifard

    2016-01-01

    Full Text Available Fungal infections are as a cause of morbidity and mortality in immunocompromise patients. Because the respiratory tract is colonized with Candida, the presence of this agent in respiratory specimens makes the diagnosis of Candida pneumonia problematic. Candida pneumonia is a rare infection, and the majority of cases are secondary to hematogenous dissemination. Furthermore isolated Candida pneumonia originating from endotracheal inoculation is an extremely rare entity. We describe a case of isolated pulmonary candidiasis in the form multiple nodular lesions in a patient with long-term history of diabetes mellitus without evidence of fungemia or systemic involvement who responded to antifungal therapy.

  15. Virulence and antifungal therapy of murine disseminated infection by Rhodotorula mucilaginosa.

    Science.gov (United States)

    Thomson, Pamela; López-Fernández, Loida; Guarro, Josep; Capilla, Javier

    2017-09-01

    Rhodotorula infections have emerged in recent years causing mainly fungemia associated to high mortality. We have evaluated the in vitro activity of nine antifungal drugs against four clinical strains of Rhodotorula mucilaginosa, being amphotericin B, voriconazole and posaconazole the most active compounds. The experimental virulence of this fungus and the efficacy of the three mentioned drugs were evaluated in disseminated infections in neutropenic mice. Infection resulted in a high fungal load in all the organs studied without evident particular tropism. All treated animals showed reduced burden respect to the control in a strain dependent manner being voriconazole slightly superior to posaconazole and amphotericin B. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Intractable urinary tract infection in a renal transplant recipient

    International Nuclear Information System (INIS)

    Gokulnath, Renuka Satish

    2009-01-01

    Urinary tract infections (UTI) are the most common bacterial infections after renal transplantation and are associated with significant morbidity and mortality. Recurrent or relapsing infections are not uncommon in the early post-transplant period and superadded fungal UTI can occur in these patients, posing a difficult therapeutic problem. Literature on recurrent UTI after transplant as well as the ideal approach to such patients is scanty. We present the case of a renal allograft recipient who presented with relapsing bacterial UTI complicated by systemic fungemia; also, a brief review of fungal UTI is attempted. (author)

  17. Glucosylceramide Administration as a Vaccination Strategy in Mouse Models of Cryptococcosis.

    Directory of Open Access Journals (Sweden)

    Visesato Mor

    Full Text Available Cryptococcus neoformans is an opportunistic fungal pathogen and the causative agent of the disease cryptococcosis. Cryptococcosis is initiated as a pulmonary infection and in conditions of immune deficiency disseminates to the blood stream and central nervous system, resulting in life-threatening meningoencephalitis. A number of studies have focused on the development of a vaccine against Cryptococcus, primarily utilizing protein-conjugated components of the Cryptococcus polysaccharide capsule as antigen. However, there is currently no vaccine against Cryptococcus in the clinic. Previous studies have shown that the glycosphingolipid, glucosylceramide (GlcCer, is a virulence factor in C. neoformans and antibodies against this lipid inhibit fungal growth and cell division. In the present study, we have investigated the possibility of using GlcCer as a therapeutic agent against C. neoformans infections in mouse models of cryptococcosis. GlcCer purified from a non-pathogenic fungus, Candida utilis, was administered intraperitoneally, prior to infecting mice with a lethal dose of C. neoformans. GlcCer administration prevented the dissemination of C. neoformans from the lungs to the brain and led to 60% mouse survival. GlcCer administration did not cause hepatic injury and elicited an anti-GlcCer antibody response, which was observed independent of the route of administration and the strains of mouse. Taken together, our results suggest that fungal GlcCer can protect mice against lethal doses of C. neoformans infection and can provide a viable vaccination strategy against Cryptococcus.

  18. Glucosylceramide Administration as a Vaccination Strategy in Mouse Models of Cryptococcosis.

    Science.gov (United States)

    Mor, Visesato; Farnoud, Amir M; Singh, Ashutosh; Rella, Antonella; Tanno, Hiromasa; Ishii, Keiko; Kawakami, Kazuyoshi; Sato, Toshiya; Del Poeta, Maurizio

    2016-01-01

    Cryptococcus neoformans is an opportunistic fungal pathogen and the causative agent of the disease cryptococcosis. Cryptococcosis is initiated as a pulmonary infection and in conditions of immune deficiency disseminates to the blood stream and central nervous system, resulting in life-threatening meningoencephalitis. A number of studies have focused on the development of a vaccine against Cryptococcus, primarily utilizing protein-conjugated components of the Cryptococcus polysaccharide capsule as antigen. However, there is currently no vaccine against Cryptococcus in the clinic. Previous studies have shown that the glycosphingolipid, glucosylceramide (GlcCer), is a virulence factor in C. neoformans and antibodies against this lipid inhibit fungal growth and cell division. In the present study, we have investigated the possibility of using GlcCer as a therapeutic agent against C. neoformans infections in mouse models of cryptococcosis. GlcCer purified from a non-pathogenic fungus, Candida utilis, was administered intraperitoneally, prior to infecting mice with a lethal dose of C. neoformans. GlcCer administration prevented the dissemination of C. neoformans from the lungs to the brain and led to 60% mouse survival. GlcCer administration did not cause hepatic injury and elicited an anti-GlcCer antibody response, which was observed independent of the route of administration and the strains of mouse. Taken together, our results suggest that fungal GlcCer can protect mice against lethal doses of C. neoformans infection and can provide a viable vaccination strategy against Cryptococcus.

  19. Fungal genome and mating system transitions facilitated by chromosomal translocations involving intercentromeric recombination.

    Directory of Open Access Journals (Sweden)

    Sheng Sun

    2017-08-01

    Full Text Available Species within the human pathogenic Cryptococcus species complex are major threats to public health, causing approximately 1 million annual infections globally. Cryptococcus amylolentus is the most closely known related species of the pathogenic Cryptococcus species complex, and it is non-pathogenic. Additionally, while pathogenic Cryptococcus species have bipolar mating systems with a single large mating type (MAT locus that represents a derived state in Basidiomycetes, C. amylolentus has a tetrapolar mating system with 2 MAT loci (P/R and HD located on different chromosomes. Thus, studying C. amylolentus will shed light on the transition from tetrapolar to bipolar mating systems in the pathogenic Cryptococcus species, as well as its possible link with the origin and evolution of pathogenesis. In this study, we sequenced, assembled, and annotated the genomes of 2 C. amylolentus isolates, CBS6039 and CBS6273, which are sexual and interfertile. Genome comparison between the 2 C. amylolentus isolates identified the boundaries and the complete gene contents of the P/R and HD MAT loci. Bioinformatic and chromatin immunoprecipitation sequencing (ChIP-seq analyses revealed that, similar to those of the pathogenic Cryptococcus species, C. amylolentus has regional centromeres (CENs that are enriched with species-specific transposable and repetitive DNA elements. Additionally, we found that while neither the P/R nor the HD locus is physically closely linked to its centromere in C. amylolentus, and the regions between the MAT loci and their respective centromeres show overall synteny between the 2 genomes, both MAT loci exhibit genetic linkage to their respective centromere during meiosis, suggesting the presence of recombinational suppressors and/or epistatic gene interactions in the MAT-CEN intervening regions. Furthermore, genomic comparisons between C. amylolentus and related pathogenic Cryptococcus species provide evidence that multiple chromosomal

  20. Primær kutan kryptokokkose hos dueavler

    DEFF Research Database (Denmark)

    Andersen, Kjeld; Ravn, Pernille; Bangsborg, Jette Marie

    2011-01-01

    A Danish 73 year-old male patient with polymyalgia rheumatica and in prednisone treatment presented with a cutaneous ulcer on the hand caused by a scratch from a pigeon. Initial treatment with antibiotics was unsuccessful. Biopsy revealed Cryptococcus neoformans and the patient recovered on antif......A Danish 73 year-old male patient with polymyalgia rheumatica and in prednisone treatment presented with a cutaneous ulcer on the hand caused by a scratch from a pigeon. Initial treatment with antibiotics was unsuccessful. Biopsy revealed Cryptococcus neoformans and the patient recovered...

  1. Cryptococcal osteomyelitis: a report of 5 cases and a review of the recent literature.

    Science.gov (United States)

    Medaris, Leigh Ann; Ponce, Brent; Hyde, Zane; Delgado, Dennis; Ennis, David; Lapidus, William; Larrison, Matthew; Pappas, Peter G

    2016-06-01

    Cryptococcus neoformans is a fungal pathogen associated with advanced HIV disease and other disorders associated with immune dysfunction. The pulmonary and the central nervous system are the most common manifestations of the disease. Localised osteomyelitis as the sole manifestation of extrapulmonary disease is rare. Herein, we present five cases of Cryptococcus osteomyelitis as the only manifestation of extrapulmonary disease. We also identified 84 additional cases of isolated cryptococcal osteomyelitis in the literature. Using these data, we have made some general recommendations regarding an approach to treatment of this uncommon clinical entity. © 2016 Blackwell Verlag GmbH.

  2. Epidemiology of Rhodotorula: An Emerging Pathogen

    Science.gov (United States)

    Wirth, Fernanda; Goldani, Luciano Z.

    2012-01-01

    This is an updated paper focusing on the general epidemiological aspects of Rhodotorula in humans, animals, and the environment. Previously considered nonpathogenic, Rhodotorula species have emerged as opportunistic pathogens that have the ability to colonise and infect susceptible patients. Rhodotorula species are ubiquitous saprophytic yeasts that can be recovered from many environmental sources. Several authors describe the isolation of this fungus from different ecosystems, including sites with unfavourable conditions. Compared to R. mucilaginosa, R. glutinis and R. minuta are less frequently isolated from natural environments. Among the few references to the pathogenicity of Rhodotorula spp. in animals, there are several reports of an outbreak of skin infections in chickens and sea animals and lung infections and otitis in sheep and cattle. Most of the cases of infection due to Rhodotorula in humans were fungemia associated with central venous catheter (CVC) use. The most common underlying diseases included solid and haematologic malignancies in patients who were receiving corticosteroids and cytotoxic drugs, the presence of CVC, and the use of broad-spectrum antibiotics. Unlike fungemia, some of the other localised infections caused by Rhodotorula, including meningeal, skin, ocular, peritoneal, and prosthetic joint infections, are not necessarily linked to the use of CVCs or immunosuppression. PMID:23091485

  3. Rapid identification of moulds and arthroconidial yeasts from positive blood cultures by MALDI-TOF mass spectrometry.

    Science.gov (United States)

    de Almeida, João N; Sztajnbok, Jaques; da Silva, Afonso Rafael; Vieira, Vinicius Adriano; Galastri, Anne Layze; Bissoli, Leandro; Litvinov, Nadia; Del Negro, Gilda Maria Barbaro; Motta, Adriana Lopes; Rossi, Flávia; Benard, Gil

    2016-11-01

    Moulds and arthroconidial yeasts are potential life-threatening agents of fungemia in immunocompromised patients. Fast and accurate identification (ID) of these pathogens hastens initiation of targeted antifungal therapy, thereby improving the patients' prognosis. We describe a new strategy that enabled the identification of moulds and arthroconidial yeasts directly from positive blood cultures by MALDI-TOF mass spectrometry (MS). Positive blood cultures (BCs) with Gram staining showing hyphae and/or arthroconidia were prospectively selected and submitted to an in-house protein extraction protocol. Mass spectra were obtained by Vitek MS™ system, and identifications were carried out with in the research use only (RUO) mode with an extended database (SARAMIS™ [v.4.12] plus in-house database). Fusarium solani, Fusarium verticillioides, Exophiala dermatitidis, Saprochaete clavata, and Trichosporon asahii had correct species ID by MALDI-TOF MS analysis of positive BCs. All cases were related to critically ill patients with high mortality fungemia and direct ID from positive BCs was helpful for rapid administration of targeted antifungal therapy. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. 288-IJBCS-Article-Dr Tene Mathieu

    African Journals Online (AJOL)

    Dr Gatsing

    active antibacterial and antifungal principles supports the use of C. macrostachys in traditional medicine for the treatment of microbial infections. .... Virginia, USA), in addition to one fungus,. Cryptococcus neoformans IP 95026, from. Pasteur ... attention as far back as in the 19th century. (Tolstikova et al., 2006). For all the ...

  5. New secondary metabolites from bioactive extracts of the fungus Armillaria tabescens

    Science.gov (United States)

    H. M. T.Bandara Herath; Melissa Jacob; A. Alpus Wilson; Hamed K. Abbas; N.P. Dhammika Nanayakkara. Nanayakkara

    2012-01-01

    Ethyl acetate extracts of Armillaria tabescens (strain JNB-OZ344) showed significant fungistatic and bacteristatic activities against several major human pathogens including Candida albicans, Cryptococcus neoformans, Escherichia coli and Mycobacterium intracellulare. Chemical analysis of these extracts led to the isolation and identification of four new compounds,...

  6. Beech Bark Disease

    Science.gov (United States)

    David R. Houston; James T. O' Brien

    1983-01-01

    Beech bark disease causes significant mortality and defect in American beech, Fagus grandifolia (Ehrh.). The disease results when bark, attacked and altered by the beech scale, Cryptococcus fagisuga Lind., is invaded and killed by fungi, primarily Nectria coccinea var. faginata Lohman, Watson, and Ayers, and sometimes N. galligena Bres.

  7. In vitro antifungal activity of methanol extracts of some Indian ...

    African Journals Online (AJOL)

    The methanol extract of 9 Indian medicinal plants belonging to 9 different families were evaluated for in vitro antifungal activity against some yeasts including Candida albicans (1) ATCC2091, C. albicans (2) ATCC18804, Candida glabrata NCIM3448, Candida tropicalis ATCC4563, Cryptococcus luteolus ATCC32044, ...

  8. EAMJ April Profile.indd

    African Journals Online (AJOL)

    2009-04-04

    Apr 4, 2009 ... niger, Fusarium, Rhizopus, Altenaria spp, Candida spp and Cryptococcus were isolated. Conclusion: Money coins harbour potentially pathogenic bacteria and fungi that may pose a public health risk. Hand hygiene is therefore strongly recommended, especially for those who simultaneously handle food ...

  9. Outcome of HIV-1-associated cryptococcal meningitis, Denmark 1988-2008

    DEFF Research Database (Denmark)

    Mathiesen, Inger Hee Mabuza; Knudsen, Jenny Dahl; Gerstoft, Jan

    2012-01-01

    Abstract Introduction: The risk of HIV-1-associated Cryptococcus neoformans meningitis (CM) has decreased and the outcome has improved with the use of combination antiretroviral therapy (cART). Outcome has not been reported in Denmark in the cART era. Methods: A review of all cases of HIV-1...

  10. Cryptococcal Meningitis in a Newly Diagnosed AIDS Patient: A Case ...

    African Journals Online (AJOL)

    BACKGROUND: Cryptococcus neoformans is a very important cause of fungal meningitis in immunosuppressed patients. OBJECTIVE: To describe a case of cryptococcal meningoencephalitis in an HIV/AIDS patient from the University of Ilorin Teaching Hospital. METHODS: An 18 -year -old male student presented with ...

  11. Hepatic safety of itraconazole intravenous solution in treatment of invasive fungal infection

    Institute of Scientific and Technical Information of China (English)

    朱利平

    2006-01-01

    Objective To investigate the hepatic safety of itraconazole intravenous solution in the treatment of invasive fungal infection. Methods Forty-nine patients with invasive fungal infection, such as pneumonia, meningitis, endocarditis, and blood stream infection, caused by Aspergillus spp. Cryptococcus neoformans, Candida spp. Penicillium marneffei,and Prototheca wiekerhamii, 50 of which had underlying diseases, including hepatic disea-

  12. The yeast flora of some decaying mushrooms on trunks of living trees

    NARCIS (Netherlands)

    Middelhoven, W.J.

    2004-01-01

    Several ascomycetous and basidiomycetous yeasts were isolated from rotten mushrooms on the trunks of beech and tamarisk trees. One strain, identified as the novel species Cryptococcus allantoinivorans, assimilated allantoin as the sole carbon source. Phylogenetically it belongs to the C. laurentii

  13. Cryptococcal rib osteomyelitis as primary and only symptom of idiopathic CD4 penia

    DEFF Research Database (Denmark)

    Legarth, Rebecca A; Christensen, Merete; Calum, Henrik

    2014-01-01

    A 59-year old man with idiopathic CD4 lymphopenia presented with extensive disseminated Cryptococcus neoformans infection including a large rib cryptoccocoma, vertebral spondylitis and pleural empyema. Complete resection of the affected part of the rib was necessary after failure of initial antif...

  14. Criptococose em felino Cryptococcosis in cat

    Directory of Open Access Journals (Sweden)

    F.J.F. Sant’Ana

    1999-08-01

    Full Text Available A case of cryptococcosis in a cat refferred to the Hospital Veterinário da Universidade Federal Rural de Pernambuco is described. The cat was euthanized and the microscopic examination of a firm mass observed in the nasal cavity was accomplished. Cryptococcus sp. and a chronic inflammatory process was observed throughout the tissue.

  15. Hamacanthins A and B, new antifungal bis indole alkaloids from the deep-water marine sponge, Hamacantha sp.

    Science.gov (United States)

    Gunasekera, S P; McCarthy, P J; Kelly-Borges, M

    1994-10-01

    Hamacanthin A [1] and hamacanthin B [2] are two bioactive dihydropyrazinonediylbis(indole) alkaloids isolated from a new species of deep-water marine sponge, Hamacantha sp. The hamacanthins are growth inhibitors of Candida albicans and Cryptococcus neoformans. Isolation and structure elucidation of 1 and 2 by nmr spectroscopy are described.

  16. Screening of Methanol and Acetone Extracts of Fourteen Indian Medicinal Plants for Antimicrobial Activity

    OpenAIRE

    VAGHASIYA, Yogeshkumar; CHANDA, Sumitra V.

    2014-01-01

    The methanol and acetone extracts of 14 plants belonging to different families were evaluated for antimicrobial activity against five Gram-positive bacteria: Staphylococcus aureus, Staphylococcus epidermidis, Bacillus cereus, Bacillus subtilis, Micrococcus flavus; seven Gram-negative bacteria: Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Proteus vulgaris, Salmonella typhimurium, Citrobacter freundii; and three fungi: Candida tropicalis, Cryptococcus lut...

  17. Antifungal Activity of Hypericum havvae Against Some Medical ...

    African Journals Online (AJOL)

    ... potency against Candida albicans and Cryptococcus laurentii, with the same MIC value of 1.56 mg/ml. Conclusion: Our findings support the use of Hypericum havvae in traditional medicine for the treatment of fungal infections, especially Candidiasis. Keywords: Antifungal activity, Candida, Hypericum havvae, Candidiasis ...

  18. In-vitro Antimicrobial and Antitumor Activities of Stevia Rebaudiana ...

    African Journals Online (AJOL)

    subtilis, Aeromonas hydrophila and Vibrio cholerae by using agar well diffusion method. Candida albicans, Cryptococcus neoformans, Trichophyton mentagrophytes and Epidermophyton species were used to test anti-yeast and antifungal activity. The cytotoxic effects of the extracts on Vero and HEp2 cells were assayed ...

  19. Laccase-catalysed oxidations of naturally occurring phenols: from in vivo biosynthetic pathways to green synthetic applications

    Czech Academy of Sciences Publication Activity Database

    Jong-Rok, J.; Baldrian, Petr; Murugesan, K.; Chang, Y.-S.

    2012-01-01

    Roč. 5, č. 3 (2012), s. 318-332 ISSN 1751-7907 Institutional research plan: CEZ:AV0Z50200510 Keywords : POLYCYCLIC AROMATIC-HYDROCARBONS * CRYPTOCOCCUS-NEOFORMANS * LIGNIN BIOSYNTHESIS Subject RIV: EE - Microbiology, Virology Impact factor: 3.214, year: 2012

  20. Ethiopian Journal of Health Development - Vol 15, No 1 (2001)

    African Journals Online (AJOL)

    Histopathological findings of Post-Menopausal bleeding in Ethiopian women. Wondwossen Ergete, Abiye Tesfaye. http://dx.doi.org/10.4314/ejhd.v15i1.9895 · Isolation and characterization of Cryptococcus neoformans from environmental sources in Ethiopia. Yimtubezenash W. Amanuel, Leykun Jemaneh, Dawit Abate.

  1. Cryptococcal pyarthrosis and sarcoidosis

    Energy Technology Data Exchange (ETDEWEB)

    Geller, David S.; Pope, John B. [Albert Einstein College of Medicine, Montefiore Medical Center, Department of Orthopaedic Surgery, Bronx, NY (United States); Thornhill, Beverly A. [Albert Einstein College of Medicine, Montefiore Medical Center, Department of Radiology, Musculoskeletal Radiology, Bronx, NY (United States); Dorfman, Howard D. [Albert Einstein College of Medicine, Montefiore Medical Center, Section of Orthopaedic Pathology, Bronx, NY (United States)

    2009-07-15

    Cryptococcus neoformans is an infrequent cause of septic arthritis. Cryptococcal infections have been linked to sarcoidosis because of both inherent immunologic consequences of the disease and its typical immune modulating treatments. Cryptococcal infections should be suspected in patients with underlying immune deficiencies, and a high degree of vigilance should be exercised to avoid misdiagnosis, dissemination of infection, and meningitis. (orig.)

  2. Screening for resistance to beech bark disease: Improvements and results from seedlings and grafted field selections

    Science.gov (United States)

    Jennifer L. Koch; Mary E. Mason; David W. Carey

    2012-01-01

    Beech bark disease (BBD) is an insect-disease complex that has been killing American beech (Fagus grandifolia Ehrh.) trees since the accidental introduction of the beech scale insect (Cryptococcus fagisuga) to Canada around 1890. Insect infestation is followed by infection with Neonectria ditissima or

  3. Development of molecular tools for use in beech bark disease management

    Science.gov (United States)

    Jennifer L. Koch; David W. Carey; Mary E. Mason; C. Dana Nelson; Abdelali Barakat; John E. Carlson; David. Neale

    2011-01-01

    Beech bark disease (BBD) has been killing American beech trees in eastern North America since the late 1890s. The disease is initiated by feeding of the beech scale insect, Cryptococcus fagisuga, which leads to the development of small fissures in the bark.

  4. Assessment of beech scale resistance in full- and half-sibling American beech families

    Science.gov (United States)

    Jennifer L. Koch; David W. Carey; Mary E. Mason; C. Dana Nelson

    2010-01-01

    A beech bark disease infested American beech tree (Fagus grandifolia Ehrh.) and two uninfested trees were selected in a mature natural stand in Michigan, USA, and mated to form two full-sib families for evaluating the inheritance of resistance to beech scale (Cryptococcus fagisuga Lind.), the insect element of beech bark disease....

  5. Adventitious sporulation in Fusarium: The yeast that were not

    Directory of Open Access Journals (Sweden)

    Matthew B. Lockwood

    2016-01-01

    Full Text Available In immunocompromised patients, Fusarium species cause infections that lead to high mortality. Our case report describes a case of disseminated fusariosis in a neutropenic patient with AML after myelosuppressive chemotherapy, and a neutropenic multiple myeloma patient with Fusarium fungemia awaiting stem cell collection. Both cases highlight the fact that Fusarium can grow as yeast-like structures in the blood causing a delay in diagnosis, and that Fusarium has a tendency to be a resistant organism. Fusarium was only susceptible to amphotericin B in both cases, but we chose to continue treatment with voriconazole in the first case with disseminated infection, despite culture results, in view of his good clinical response. Despite high mortality rates in disseminated infection, our two patients had good outcomes.

  6. Utilidade do coágulo sangüíneo para o isolamento de Sporothrix schenckii de gatos naturalmente infectados

    Directory of Open Access Journals (Sweden)

    Tânia Maria Pacheco Schubach

    2004-11-01

    Full Text Available The diagnosis of disseminated sporotrichosis is usually obtained by necropsy and the isolation of Sporothrix schenckii from blood is rare. Fungemia was shown in vivo through the isolation of S. schenckii from peripheral blood of 13 (n=38, 34.2% cats with naturally acquired sporotrichosis. The coinfection with FIV and with FeLV found, respectively, in 6 (n=34, 17.6% cases and 1 (n=34, 2.9%, apparently did not alter the frequency of the isolation of S. schenckii from peripheral blood. There was agreement of 84.2% comparing these results to the blood culture results simultaneously achieved. In this way, we propose the clot culture as a practical alternative method, efficient and cheap for the diagnosis of disseminated sporotrichosis in cats in vivo.

  7. MALDI-TOF mass spectrometry in the clinical mycology laboratory: identification of fungi and beyond.

    Science.gov (United States)

    Posteraro, Brunella; De Carolis, Elena; Vella, Antonietta; Sanguinetti, Maurizio

    2013-04-01

    MALDI-TOF mass spectrometry (MS) is becoming essential in most clinical microbiology laboratories throughout the world. Its successful use is mainly attributable to the low operational costs, the universality and flexibility of detection, as well as the specificity and speed of analysis. Based on characteristic protein spectra obtained from intact cells - by means of simple, rapid and reproducible preanalytical and analytical protocols - MALDI-TOF MS allows a highly discriminatory identification of yeasts and filamentous fungi starting from colonies. Whenever used early, direct identification of yeasts from positive blood cultures has the potential to greatly shorten turnaround times and to improve laboratory diagnosis of fungemia. More recently, but still at an infancy stage, MALDI-TOF MS is used to perform strain typing and to determine antifungal drug susceptibility. In this article, the authors discuss how the MALDI-TOF MS technology is destined to become a powerful tool for routine mycological diagnostics.

  8. [Urinary infection by Saccharomyces cerevisiae: Emerging yeast?].

    Science.gov (United States)

    Elkhihal, B; Elhalimi, M; Ghfir, B; Mostachi, A; Lyagoubi, M; Aoufi, S

    2015-12-01

    Saccharomyces cerevisiae is a commensal yeast of the digestive, respiratory and genito-urinary tract. It is widely used as a probiotic for the treatment of post-antibiotic diarrhea. It most often occurs in immunocompromised patients frequently causing fungemia. We report the case of an adult diabetic patient who had a urinary tract infection due to S. cerevisiae. The disease started with urination associated with urinary frequency burns without fever. The diagnosis was established by the presence of yeasts on direct examination and positivity of culture on Sabouraud-chloramphenicol three times. The auxanogramme gallery (Auxacolor BioRad(®)) allowed the identification of S. cerevisiae. The patient was put on fluconazole with good outcome. This observation points out that this is an opportunistic yeast in immunocompromised patients. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  9. Refractory Post-Herpetic Neuralgia As An Initial Presentation Of Olfactory Neuroblastoma-Related Ectopic ACTH Syndrome

    Directory of Open Access Journals (Sweden)

    Hsiang-Hung Lin

    2009-03-01

    Full Text Available We report a woman aged 64 years with ectopic adrenocorticotropic hormone (ACTH syndrome caused by olfactory neuroblastoma as an initial presentation of refractory post-herpetic neuralgia. The manifestations such as cushingoid appearance and endocrine abnormalities are compatible with Cushing's syndrome. Brain computed tomography revealed a sellar mass. A biopsy revealed olfactory neuroblastoma. Immunohistochemical staining further defined the tumor as an ACTH-secreting neuroblastoma. Subsequent opportunistic infections by Candida glabrata fungemia and multiple drug-resistant Acinetobacter baumannii pneumonia occurred during hospitalization as a complication of severe hypercortisolism. Before any therapy for Cushing's syndrome and neuroblastoma could be initiated, the patient died from sepsis and multiorgan failure. We propose that Cushing's syndrome is more complex than what clinicians thought, and that meticulous cerebral imaging studies are crucial.

  10. Endocarditis due to Rhodotorula mucilaginosa in a kidney transplanted patient: case report and review of medical literature

    Science.gov (United States)

    Cabral, Andrea Maria; da Siveira Rioja, Suzimar; Brito-Santos, Fabio; Peres da Silva, Juliana Ribeiro; MacDowell, Maria Luíza; Melhem, Marcia S. C.; Mattos-Guaraldi, Ana Luíza; Hirata Junior, Raphael

    2017-01-01

    Introduction. Endocarditis caused by yeasts is currently an emerging cause of infective endocarditis and, when accompanied byfever of unknown origin, is more severe since interferes with proper diagnosis and endocarditis treatment. Case presentation. The Rio de Janeiro Infective Endocarditis Study Group reports a case of infectious endocarditis (IE) with negative blood cultures in a 45-year-old white female resident in Rio de Janeiro, Brazil, previously submitted to kidney transplantation. After diagnosis and intervention, the valve culture revealed Rhodotorula mucilaginosa. The clinical aspects and overview of endocarditis caused by Rhodotorula spp. demonstrated that R. muscilaginosa have been isolated from the last IE cases from kidney transplanted patients. Conclusion. Though most of the patients (in literature) recovered well from endocarditis caused by Rhodotorula spp., physicians must be aware for diagnosis of fungemia and fungal treatment in kidney transplanted patients suffering of fever of unknown origin in the modern immunosuppressive treatment. PMID:29255609

  11. Itraconazole-resistant Candida auris with phospholipase, proteinase and hemolysin activity from a case of vulvovaginitis.

    Science.gov (United States)

    Kumar, Dharmendra; Banerjee, Tuhina; Pratap, Chandra Bhan; Tilak, Ragini

    2015-04-15

    Since the emergence of pathogenic non-albicans Candida species, a number of new isolates have been added to the list. One such unusual species is Candida auris (C. auris), recently isolated and studied in few reports. In this study, a case of vulvovaginitis caused by Candida auris incidentally identified by molecular methods using internal transcribed spacer polymerase chain reaction (ITS PCR) is described. Antifungal susceptibility testing revealed the isolate to be resistant to itraconazole (MIC ≥ 2 µg/ml) and expressed important virulence factors including phospholipase, proteinase and hemolysin activity. The patient was successfully treated with oral fluconazole and did not have any invasive fungemia. Very few cases of this emerging pathogen have been reported. However, its isolation from clinical specimens reveals the significance of non-albicans candida species over C. albicans and the diversity of Candida spp causing infections.

  12. Mycotic aneurysm of the superior mesenteric artery after septic embolism

    International Nuclear Information System (INIS)

    Schmidt, F.; Dinkel, H.P.

    2002-01-01

    Mycotic aneurysms of the aorta and the visceral arteries are life-threatening diseases, due to potential rupture and organ or limb ischemia. They occur in endocarditis, immunodeficiency, bacteremia and fungemia, and have a poor prognosis.We report on a case of a 54-year-old male patient suffering from abdominal angina after mitral valve replacement for septic mycotic endocarditis. In presence of a mycotic-embolic occlusion of the left popliteal artery and multiple septic organ infarctions a mycotic aneurysm of the superior mesenteric artery was found in abdominal spiral-CT.Based on sequential spiral-CT examinations, this case demonstrates the development of a septic aneurysm of the superior mesenteric artery. (orig.) [de

  13. Outcomes, impact on management, and costs of fungal eye disease consults in a tertiary care setting.

    Science.gov (United States)

    Ghodasra, Devon H; Eftekhari, Kian; Shah, Ankoor R; VanderBeek, Brian L

    2014-12-01

    To determine the frequency of clinical management changes resulting from inpatient ophthalmic consultations for fungemia and the associated costs. Retrospective case series. Three hundred forty-eight inpatients at a tertiary care center between 2008 and 2012 with positive fungal blood culture results, 238 of whom underwent an ophthalmologic consultation. Inpatient charts of all fungemic patients were reviewed. Costs were standardized to the year 2014. The Student t test was used for all continuous variables and the Pearson chi-square test was used for categorical variables. Prevalence of ocular involvement, rate of change in clinical management, mortality rate of fungemic patients, and costs of ophthalmic consultation. Twenty-two (9.2%) of 238 consulted patients with fungemia had ocular involvement. Twenty patients had chorioretinitis and 2 had endophthalmitis. Only 9 patients (3.7%) had a change in management because of the ophthalmic consultation. One patient underwent bilateral intravitreal injections. Thirty percent of consulted patients died before discharge or were discharged to hospice. The total cost of new consults was $36 927.54 ($204.19/initial level 5 visit and $138.63/initial level 4). The cost of follow-up visits was $13 655.44 ($104.24/visit). On average, 26.4 patients were evaluated to find 1 patient needing change in management, with an average cost of $5620.33 per change in 1 patient's management. Clinical management changes resulting from ophthalmic consultation in fungemic patients were uncommon. Associated costs were high for these consults in a patient population with a high mortality rate. Together, these data suggest that the usefulness of routine ophthalmic consultations for all fungemic patients is likely to be low. Copyright © 2014 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.

  14. Microbial ecology of extreme environments: Antarctic dry valley yeasts and growth in substrate-limited habitats

    Science.gov (United States)

    Vishniac, H. S.

    1982-01-01

    The success of the Antarctic Dry Valley yeasts presumeably results from adaptations to multiple stresses, to low temperatures and substrate-limitation as well as prolonged resting periods enforced by low water availability. Previous investigations have suggested that the crucial stress is substrate limitation. Specific adaptations may be pinpointed by comparing the physiology of the Cryptococcus vishniacii complex, the yeasts of the Tyrol Valley, with their congeners from other habitats. Progress was made in methods of isolation and definition of ecological niches, in the design of experiments in competition for limited substrate, and in establishing the relationships of the Cryptococcus vishniacii complex with other yeasts. In the course of investigating relationships, a new method for 25SrRNA homology was developed. For the first time it appears that 25SrRNA homology may reflect parallel or convergent evolution.

  15. Cryptococcal meningitis

    Directory of Open Access Journals (Sweden)

    DING Wen-ting

    2013-01-01

    Full Text Available Cryptococcus neoformans is a kind of encapsulated fungal organism that widely exists in the nature. Because of its neurotropic nature, the central nervous system becomes its major target organ. Cryptococcus neoformans can use "transcellular pathway", "paracellular pathway" and "Trojan horse approach" to cross blood-brain barrier, and then make the devastating diffusion. Despite antifungal therapy, the mortality rate remains between 10% and 25% in patients with cryptococcal meningitis (CM and acquired immunodeficiency syndrome (AIDS, and at least one-third of patients have experienced failure of antifungal therapy. Consequently, it is very important for us to understand the pathogenesis of CM, to diagnose as soon as possible and to explore more reasonable treatment.

  16. Antifungal Activity and Composition of Essential Oils of Conyza canadensis Herbs and Roots

    Directory of Open Access Journals (Sweden)

    Katalin Veres

    2012-01-01

    Full Text Available Essential oils from herbs and roots of Conyza canadensis (horseweed, collected in Hungary, were obtained by hydrodistillation. The chemical compositions of the oils were analysed by combination of GC and GC/MS. The major constituent of the oil obtained from the aerial parts of horseweed was limonene (78%, while the main component of root oil was 2Z,8Z-matricaria ester. The antimicrobial activities of the oils were tested on Gram-positive (Enterococcus faecalis, Staphylococcus aureus, and Streptococcus pyogenes, Gram-negative (Escherichia coli, Pseudomonas aeruginosa bacteria, reference fungal strains, and fungal strains isolated from patients (Candida, Cryptococcus, Trichophyton, Rhodotorula, and Aspergillus by agar disc diffusion and broth dilution methods. None of the oils showed any activity against the tested bacterial strains, but exhibited moderate-to-strong activity against all fungi with the only exception of A. fumigatus. The highest zone of inhibition was observed in case of Cryptococcus neoformans and Trichophyton interdigitalis

  17. Yeast diversity isolated from grape musts during spontaneous fermentation from a Brazilian winery.

    Science.gov (United States)

    Bezerra-Bussoli, Carolina; Baffi, Milla Alves; Gomes, Eleni; Da-Silva, Roberto

    2013-09-01

    Saccharomyces and non-Saccharomyces yeast species from a winery located in Brazil were identified by ribosomal gene-sequencing analysis. A total of 130 yeast strains were isolated from grape surfaces and musts during alcoholic fermentation from Isabel, Bordeaux, and Cabernet Sauvignon varieties. Samples were submitted to PCR-RFLP analysis and genomic sequencing. Thirteen species were identified: Candida quercitrusa, Candida stellata, Cryptococcus flavescens, Cryptococcus laurentii, Hanseniaspora uvarum, Issatchenkia occidentalis, Issatchenkia orientalis, Issatchenkia terricola, Pichia kluyveri, Pichia guilliermondii, Pichia sp., Saccharomyces cerevisiae, and Sporidiobolus pararoseus. A sequential substitution of species during the different stages of fermentation, with a dominance of non-Saccharomyces yeasts at the beginning, and a successive replacement of species by S. cerevisiae strains at the final steps were observed. This is the first report about the yeast distribution present throughout the alcoholic fermentation in a Brazilian winery, providing supportive information for future studies on their contribution to wine quality.

  18. Sertraline demonstrates fungicidal activity in vitro for Coccidioides immitis

    Directory of Open Access Journals (Sweden)

    Simon Paul

    2016-07-01

    Full Text Available Coccidioidomycosis causes substantial morbidity in endemic areas. Disseminated coccidioidomycosis is an AIDS defining condition and treatment often requires lifelong antifungal therapy. Sertraline, a widely used serotonin-reuptake inhibitor anti-depressant, has demonstrated activity against Candida and Cryptococcus sp. both in vitro and in vivo. To evaluate if sertraline has activity against Coccidioides, the minimal inhibitory concentration (MIC and minimal fungicidal concentration (MFC of sertraline for four clinical isolates of C. immitis were determined. Sertraline was observed to have an MIC range of 4–8 µg/ml and MFC also of 4–8 µg/ml for Coccidioides. These MIC and MFC results for C. immitis are similar to those reported for Cryptococcus sp. suggesting sertraline may potentially have utility for the treatment of coccidioidomycosis.

  19. Immune reconstitution syndrome in a human immunodeficiency virus infected child due to giardiasis leading to shock

    Directory of Open Access Journals (Sweden)

    Sneha Nandy

    2015-01-01

    Full Text Available Human immunodeficiency virus (HIV-associated immune reconstitution inflammatory syndrome has been reported in association with tuberculosis, herpes zoster (shingles, Cryptococcus neoformans, Kaposi′s sarcoma, Pneumocystis pneumonia, hepatitis B virus, hepatitis C virus, herpes simplex virus, Histoplasma capsulatum, human papillomavirus, and Cytomegalovirus. However, it has never been documented with giardiasis. We present a 7-year-old HIV infected girl who developed diarrhea and shock following the initiation of antiretroviral therapy, and her stool showed the presence of giardiasis.

  20. Disseminated cryptococcosis and fluconazole resistant oral candidiasis in a patient with acquired immunodeficiency syndrome (AIDS).

    Science.gov (United States)

    Kothavade, Rajendra J; Oberai, Chetan M; Valand, Arvind G; Panthaki, Mehroo H

    2010-10-28

    Disseminated cryptococcosis and recurrent oral candidiasis was presented in a-heterosexual AIDS patient. Candida tropicalis (C.tropicalis) was isolated from the oral pseudomembranous plaques and Cryptococcus neoformans (C. neoformans) was isolated from maculopapular lesions on body parts (face, hands and chest) and body fluids (urine, expectorated sputum, and cerebrospinal fluid). In vitro drug susceptibility testing on the yeast isolates demonstrated resistance to fluconazole acquired by C. tropicalis which was a suggestive possible root cause of recurrent oral candidiasis in this patient.

  1. Cryptococcosis infection among HIV patients

    Directory of Open Access Journals (Sweden)

    Zineb Tlamcani

    2016-06-01

    Full Text Available Cryptococcosis is commonly known as a central nervous system infection due to Cryptococcus neoformans. It is one of the most frequent infections in AIDS patients. Disseminated cryptococcosis appears in almost one third of these patients. In this review, we will discuss the clinical presentation of cryptococcal infections among HIV patients and various methods of diagnosis, such as India ink, latex agglutination test and culture.

  2. Rapid Identification of Pathogenic Fungi Directly from Cultures by Using Multiplex PCR

    OpenAIRE

    Luo, Guizhen; Mitchell, Thomas G.

    2002-01-01

    A multiplex PCR method was developed to identify simultaneously multiple fungal pathogens in a single reaction. Five sets of species-specific primers were designed from the internal transcribed spacer (ITS) regions, ITS1 and ITS2, of the rRNA gene to identify Candida albicans, Candida glabrata, Candida parapsilosis, Candida tropicalis, and Aspergillus fumigatus. Another set of previously published ITS primers, CN4 and CN5, were used to identify Cryptococcus neoformans. Three sets of primers w...

  3. 结核性脑膜炎与新型隐球菌脑膜炎的鉴别分析%Differential Diagnosis of Tuberculous Meningitis and Cryptoeoecus Neoformans Meningitis

    Institute of Scientific and Technical Information of China (English)

    陈劲松; 郑建明; 黄天文; 林贵喜; 姚键

    2015-01-01

    Objective:To analyze the clinical characters and prognosis of tuberculous meningitis(TBM) and cryptococcus neoformans meningitis(CNM).Method:We retrospectively analyzed the clinical data of 115 patients with tuberculous meningitis and 24 patients with cryptococcus neoformans meningitis.The relevant medical literature was reviewed.Result:Compared with cryptococcus neoformans meningitis,the average body temperature from onset to hospitalized,the quantity of cells and the level of protein and glucose in CSF of tuberculous meningitis were higher(P0.05).Conclusion:The available indicators in CSF of cryptococcus neoformans meningitis are lower than that of tuberculous meningitis,except the levels of chloride in CSF.%目的:分析新型隐球菌脑膜炎和结核性脑膜炎的不同临床特点。方法:对115例结核性脑膜炎和24例新型隐球菌脑膜炎患者的临床资料进行回顾性分析。结果:结核性脑膜炎患者入院时平均体温、脑脊液白细胞数量、脑脊液蛋白浓度、脑脊液葡萄糖浓度均明显高于新型隐球菌脑膜炎组患者(P<0.05)。结论:除氯化物外,新型隐球菌脑膜炎患者脑脊液各项指标均低于结核性脑膜炎。

  4. The yeast flora of some decaying mushrooms on trunks of living trees

    OpenAIRE

    Middelhoven, W.J.

    2004-01-01

    Several ascomycetous and basidiomycetous yeasts were isolated from rotten mushrooms on the trunks of beech and tamarisk trees. One strain, identified as the novel species Cryptococcus allantoinivorans, assimilated allantoin as the sole carbon source. Phylogenetically it belongs to the C. laurentii complex, Papiliotrema bandonii being the closest relative. Some ascomycetous strains could not be distinguished from Pichia guillermondii, but deviated considerably in rDNA sequences. In addition to...

  5. Disseminated Cryptococcal Disease in a Patient with Chronic Lymphocytic Leukemia on Ibrutinib

    OpenAIRE

    Okamoto, Koh; Proia, Laurie A.; Demarais, Patricia L.

    2016-01-01

    Cryptococcus is a unique environmental fungus that can cause disease most often in immunocompromised individuals with defective cell-mediated immunity. Chronic lymphocytic leukemia (CLL) is not known to be a risk factor for cryptococcal disease although cases have been described mainly in patients treated with agents that suppress cell-mediated immunity. Ibrutinib is a new biologic agent used for treatment of CLL, mantle cell lymphoma, and Waldenstrom’s macroglobulinemia. It acts by inhibitin...

  6. Microalgal fatty acid methyl ester a new source of bioactive compounds with antimicrobial activity

    OpenAIRE

    Arumugham Suresh; Ramasamy Praveenkumar; Ramasamy Thangaraj; Felix Lewis Oscar; Edachery Baldev; Dharumadurai Dhanasekaran; Nooruddin Thajuddin

    2014-01-01

    Objective: To evaluate fatty acid composition and the antimicrobial activity of the major fraction of fatty acid methyl ester (FAME) extracts from three microalgae collected from freshwater lakes in Theni District, Tamil Nadu, India. Methods: Antimicrobial study was carried out by well diffusion method against bacterial as well as fungal pathogens such as Escherichia coli, Staphylococcus aureus, Enterobacter sp., Klebsiella sp., Salmonella typhi, Fusarium sp., Cryptococcus sp.,...

  7. THE COORDINATION COMPOUNDS OF COBALT (II, III) WITH DITHIOCARBAMIC ACID DERIVATIVES — MODIFICATORS OF HYDROLYTIC ENZYMES ACTIVITY

    OpenAIRE

    L. D. Varbanets; О. V. Matselyukh; N. А. Nidyalkova; Е. V. Аvdiyuk; А. V. Gudzenko; I. I. Seifullina; G. N. Маsаnоvets; N. V. Khitrich

    2013-01-01

    Chloride, bromide and isothiocyanate complexes of cobalt(II) with N-substituted thiocarbamoyl-N?-pentamethylenesulfenamides (1)–(12), and also complexes of cobalt(II, Ш) with derivatives of morpholine-4-carbodithioic acid (13)–(18) have been used as modificators of enzymes of hydrolytic action — Bacillus thurin-giensis ІМВ В-7324 peptidases, Bacillus subtilis 147 and Aspergillus flavus var. oryzae 80428 amylases, Eupenicillium erubescens 248 and Cryptococcus albidus 1001 rhamnosidases. It was...

  8. Acute lethal toxicity following passive immunization for treatment of murine cryptococcosis.

    OpenAIRE

    Savoy, A C; Lupan, D M; Manalo, P B; Roberts, J S; Schlageter, A M; Weinhold, L C; Kozel, T R

    1997-01-01

    Passive immunization with monoclonal antibodies (MAbs) specific for the major capsular polysaccharide of Cryptococcus neoformans alters the course of murine cryptococcosis. During studies of passive immunization for treatment of murine cryptococcosis, we noted the occurrence of an acute, lethal toxicity. Toxicity was characterized by scratching, lethargy, respiratory distress, collapse, and death within 20 to 60 min after injection of antibody. The toxic effect was observed only in mice with ...

  9. Differential Activation of Peritoneal Cells by Subcutaneous Treatment of Rats with Cryptococcal Antigens▿

    OpenAIRE

    Baronetti, José L.; Chiapello, Laura S.; Garro, Ana P.; Masih, Diana T.

    2009-01-01

    Previous studies in our laboratory have shown that the subcutaneous pretreatment of rats with heat-killed cells (HKC) of Cryptococcus neoformans emulsified in complete Freund adjuvant (CFA) promotes protective immunity against an intraperitoneal challenge with C. neoformans. In contrast, subcutaneous treatment with the capsular polysaccharide (PSC) emulsified in CFA exacerbates the cryptococcal infection. The purpose of this study was to analyze the mechanisms involved in these phenomena. Adh...

  10. Abdominal cryptococcosis in two dogs: diagnosis and medical management.

    Science.gov (United States)

    Tangeman, Lindsay; Davignon, Danielle; Patel, Reema; Littman, Meryl

    2015-01-01

    Canine cryptococcosis cases are typically reported as neurologic, disseminated, or both. There have been few reports of other parenchymal organ involvement. Dogs infected with Cryptococcus spp. are likely to develop central nervous system involvement, and those that are severely affected are treated aggressively with surgery and/or amphotericin B. This report describes two cases of canine abdominal cryptococcosis: one boxer with primary alimentary cryptococcosis alone and one miniature schnauzer with pancreatic and disseminated cryptococcosis. The boxer is unique in that the dog suffered from primary alimentary cryptococcosis without dissemination, secondary anemia due to gastrointestinal losses, and is the second case to have Cryptococcus spp. identified on fecal examination as part of the diagnostic workup. Unlike previous reports, surgery was not performed in either case, and both dogs were treated with fluconazole alone. Currently, both dogs are free from clinical signs, and Cryptococcus spp. antigen titers are negative at 17 and 15 mo after initial presentation. These cases suggest fluconazole may be effective therapy alone for canine abdominal cryptococcosis, negating the need for high-risk therapy options such as surgery and/or amphotericin B in some cases.

  11. The effects of chloramphenicol and 60Co irradiation on the experimental mycosis in mice inoculated intragastrically with two kinds of pathogenic yeast-like fungi

    International Nuclear Information System (INIS)

    Kitamura, Isamu

    1975-01-01

    Two kinds of pathogenic yeast, Cryptococcus neoformans and Candida albicans were inoculated into the stomachs of mice. The effects of intragastric administration of chloramphenicol (CM) and systemic irradiation with 60 Co on the acceralation of infection were studied in intragastrically inoculated mice. When Cryptococcus neoformans were simply inoculated, it was difficult for them to reside in the alimentary canal to cause infection. In case of candida albicans inoculated simply, they were also unable to induce infection though they remained in the alimentary canal for a long time. In the experiment combining intragastric administration of the fungi and CM some mice contracted the disease and died. The effect of CM in causing infection was not proved. In the experiments combining 60 Co irradiation and intragastric administration of CM and fungi, a high incidence of fungus infection was noted with both Cryptococcus neoformans and Candida albicans. Fungus lesions were observed in many organs, and in all of the mice which died systemic fungus lesions were noted. It was shown that systemic irradiation with 60 Co had a specific effect on facilitating the infection. In these studies no difference was observed in the incidence of the fungus disease in respect of the origin of the causative fungi. (J.P.N.)

  12. Silver nanoparticle production by the fungus Fusarium oxysporum: nanoparticle characterisation and analysis of antifungal activity against pathogenic yeasts

    Directory of Open Access Journals (Sweden)

    Kelly Ishida

    2014-04-01

    Full Text Available The microbial synthesis of nanoparticles is a green chemistry approach that combines nanotechnology and microbial biotechnology. The aim of this study was to obtain silver nanoparticles (SNPs using aqueous extract from the filamentous fungus Fusarium oxysporum as an alternative to chemical procedures and to evaluate its antifungal activity. SNPs production increased in a concentration-dependent way up to 1 mM silver nitrate until 30 days of reaction. Monodispersed and spherical SNPs were predominantly produced. After 60 days, it was possible to observe degenerated SNPs with in additional needle morphology. The SNPs showed a high antifungal activity against Candida and Cryptococcus , with minimum inhibitory concentration values ≤ 1.68 µg/mL for both genera. Morphological alterations of Cryptococcus neoformans treated with SNPs were observed such as disruption of the cell wall and cytoplasmic membrane and lost of the cytoplasm content. This work revealed that SNPs can be easily produced by F. oxysporum aqueous extracts and may be a feasible, low-cost, environmentally friendly method for generating stable and uniformly sized SNPs. Finally, we have demonstrated that these SNPs are active against pathogenic fungi, such as Candida and Cryptococcus .

  13. [Retrospective study of neuromeningeal cryptococcosis in patients infected with HIV in the infectious diseases unit of university hospital of Casablanca, Morocco].

    Science.gov (United States)

    Dollo, I; Marih, L; El Fane, M; Es-Sebbani, M; Sodqi, M; Oulad Lahsen, A; Chakib, A; El Kadioui, F; Hamdani, A; El Mabrouki, M J; Soussi Abdallaoui, M; Karima, Z; Hassoune, S; Maaroufi, A; Marhoum El Filali, K

    2016-12-01

    To report the cases of neuromeningeal cryptococcosis and to describe the clinical, paraclinical, therapeutic and outcomes of patients. Retrospective study of 43 patients infected with HIV admitted from January first 2010 to June 30th 2015 in the infectious disease unit of UHC Ibn Rochd, for neuromeningeal cryptococcus. The mean frequency of neuromeningeal cryptococcosis in patients infected with HIV was 1.4%. The mean age was 39 years and a sex ratio of 1.38. The mean CD4 count was 70 cells/mm 3 . The diagnosis of HIV was revealed by neuromeningeal cryptococcus in 77% of cases. Fifteen days interval was reported between the first symptom and hospital admission. Headache (77%) was the most represented clinical sign. The cerebrospinal fluid analysis showed hypoglycorachy (67%), hyperproteinorachy (65%) and lymphocytosis (63%). Chinese ink direct examination for Cryptococcus neoformans in CSF was positive in 86% of cases and all cases were positive after culture on Sabouraud's medium. Patients were treated with monotherapy amphotericin B (42%) or fluconazole (28%) and bitherapy amphotéricine B/fluconazole (28%). Fatal evolution was observed in 60% of cases. Neuromeningeal cryptococcosis remains a severe opportunistic infection in HIV patients with a heavy mortality rate. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  14. Biomass production by Antarctic yeast strains: an investigation on the lipid composition

    International Nuclear Information System (INIS)

    Zlatanov, M.; Antova, G.; Angelova-Romova, M.; Pavlova, K.; Georgieva, K.; Rousenova-Videva, S.

    2010-01-01

    Psychrophilic yeast strains Rhodotorula glutinis AL_1_0_7, Sporobolomyces roseus AL_1_0_8, Cryptococcus albidus AL_5_5, Cryptococcus laurentii AL_5_6 and Cryptococcus laurentii AL_5_8 isolated from soil sample taken from the region of the Bulgarien base on Livingston Island, Antarctica, were studied. The biomass production was followed after cultivation of the yeasts in a medium with pH 5.3 at 15°C for 120 h. The biomass concentration by psychrophilic yeast strains was: R. glutinis AL_1_0_7-6.05 g/l, S. roseus AL108-5.78 g/l, Cr. albidus AL_5_5, Cr. laurentii AL_5_6 and Cr. laurentii AL_5_8-6.52 g/l, 6.84 g/l and 6.24 g/l, respectively. The extracted and separated lipids from the samples were supplied to analysis and the compositions of fatty acids, phospholipids, sterols as well as tocopherols were determined. Unsaturated fatty acids, mainly oleic (58.6-63.5%) and of saturated palmitic (18.2-24.5%), predominated in triacylglycerols. Sterols (0.1-0.3%) were valued in the dry yeast biomass. The content of phospholipids, mainly phosphatidylcholine, phosphatidylinositole and phosphatidylethanolamine was found to be in the range of 0.2-1.6%. The quantity of tocopherols was 0-26.3 mg/kg. All of tocopherol classes were established.

  15. Fluconazole levels in serum and cerebrospinal fluid according to daily dosage in patients with cryptococcosis and other fungal infections

    Directory of Open Access Journals (Sweden)

    Letícia Aparecida Schiave

    2018-01-01

    Full Text Available Fluconazole is extensively used for the treatment of candidiasis and cryptococcosis. Among other factors, successful treatment is related to appropriate fluconazole levels in blood and cerebrospinal fluid. In the present study, fluconazole levels were determined in 15 patients, 14 of whom had AIDS and 13 had neurocryptococcosis. The only selection criterion was treatment with fluconazole, which was performed with a generic or similar form of the drug. Fluconazole level was determined by high performance liquid chromatography and the susceptibility profile of Cryptococcus spp. isolated from the patients was assessed by broth microdilution. Blood and cerebrospinal fluid fluconazole levels were found to be related to the fluconazole daily dose, and exceeded the minimum inhibitory concentration of this antifungal for the Cryptococcus spp. isolates. A good correlation was observed between serum and cerebrospinal fluid drug concentration. In conclusion, treatment with non-original fluconazole under usual medical practice conditions results in appropriate blood and cerebrospinal fluid levels of the drug for inhibiting Cryptococcus spp. susceptible to this antifungal drug. The relatively common failures of neurocryptococcosis treatment appear not to be due to insufficient fluconazole levels in the cerebrospinal fluid, especially with the use of daily doses of 400–800 mg.

  16. Fluconazole levels in serum and cerebrospinal fluid according to daily dosage in patients with cryptococcosis and other fungal infections.

    Science.gov (United States)

    Schiave, Letícia Aparecida; Nascimento, Erika; Vilar, Fernando Crivelenti; de Haes, Tissiana Marques; Takayanagui, Osvaldo Massaiti; Gaitani, Cristiane Masetto de; Martinez, Roberto

    Fluconazole is extensively used for the treatment of candidiasis and cryptococcosis. Among other factors, successful treatment is related to appropriate fluconazole levels in blood and cerebrospinal fluid. In the present study, fluconazole levels were determined in 15 patients, 14 of whom had AIDS and 13 had neurocryptococcosis. The only selection criterion was treatment with fluconazole, which was performed with a generic or similar form of the drug. Fluconazole level was determined by high performance liquid chromatography and the susceptibility profile of Cryptococcus spp. isolated from the patients was assessed by broth microdilution. Blood and cerebrospinal fluid fluconazole levels were found to be related to the fluconazole daily dose, and exceeded the minimum inhibitory concentration of this antifungal for the Cryptococcus spp. isolates. A good correlation was observed between serum and cerebrospinal fluid drug concentration. In conclusion, treatment with non-original fluconazole under usual medical practice conditions results in appropriate blood and cerebrospinal fluid levels of the drug for inhibiting Cryptococcus spp. susceptible to this antifungal drug. The relatively common failures of neurocryptococcosis treatment appear not to be due to insufficient fluconazole levels in the cerebrospinal fluid, especially with the use of daily doses of 400-800mg. Copyright © 2017 Sociedade Brasileira de Infectologia. Published by Elsevier Editora Ltda. All rights reserved.

  17. Chemical composition, toxicity and larvicidal and antifungal activities of Persea americana (avocado seed extracts Composição química, toxicidade, atividade larvicida e antifúngica de extratos de semente de Persea americana (abacate

    Directory of Open Access Journals (Sweden)

    João Jaime Giffoni Leite

    2009-04-01

    Full Text Available The present study had the aim of testing the hexane and methanol extracts of avocado seeds, in order to determine their toxicity towards Artemia salina, evaluate their larvicidal activity towards Aedes aegypti and investigate their in vitro antifungal potential against strains of Candida spp, Cryptococcus neoformans and Malassezia pachydermatis through the microdilution technique. In toxicity tests on Artemia salina, the hexane and methanol extracts from avocado seeds showed LC50 values of 2.37 and 24.13mg mL-1 respectively. Against Aedes aegypti larvae, the LC50 results obtained were 16.7mg mL-1 for hexane extract and 8.87mg mL-1 for methanol extract from avocado seeds. The extracts tested were also active against all the yeast strains tested in vitro, with differing results such that the minimum inhibitory concentration of the hexane extract ranged from 0.625 to 1.25mg L-¹, from 0.312 to 0.625mg mL-1 and from 0.031 to 0.625mg mL-1, for the strains of Candida spp, Cryptococcus neoformans and Malassezia pachydermatis, respectively. The minimal inhibitory concentration for the methanol extract ranged from 0.125 to 0.625mg mL-1, from 0.08 to 0.156mg mL-1 and from 0.312 to 0.625mg mL-1, for the strains of Candida spp., Cryptococcus neoformans and Malassezia pachydermatis, respectively.O presente estudo teve como objetivo testar os extratos hexânico e metanólico das sementes do abacate, a fim de determinar sua toxicidade em Artemia salina, avaliar a atividade larvicida frente ao Aedes aegypti, bem como verificar o potencial antifúngico in vitro contra cepas de Candida spp, Cryptococcus neoformans e Malassezia pachydermatis, através da técnica de microdiluição. Os extratos hexânico e metanólico das sementes de abacate apresentaram no teste de toxicidade frente à Artemia salina, valores de LC50 2,37 e 24,13mg L-1, respectivamente; contra as larvas do Aedes aegypti os resultados obtidos foram LC50 16,7mg L-1 para o extrato hexânico e 8,87mg

  18. The Frequency Of Fungi In Doubtful Appendicitis

    Directory of Open Access Journals (Sweden)

    J Hashemi

    2006-06-01

    Full Text Available Background and Aim: While nowadays,great attainments have been achieved in curing and preventing the pathogenic fungal infections, and some how there has been reduction in the number of occurrences, the occurrences of opportunistic infections have been increased. Since the study of fungal infections in various organs (e.g.digestive system is crucial ,and because of few study were done in this field in the world, it is decided to examine the apendectomide tissue for fungal contamination in Iran. Materials and Methods: The work has been done for six months. After oparation sergery the appendix tissue in two media (formalin & normal salin were carried out in the medical mycology laboratory at Tehran University of medical sciences. The specimens were examined directly and cultured in sabourauds dextrose agar with chloramphenicol (sc. In this experiment 200 appendicular tissues were examined. Results: Out of them some fungi were isolated in 10 cases included 4 Candida albican (40%, 2 Candida tropicalis (20%,1 Cryptococcus sp. (10%,1 Candida sp.and 2 Geotrichum sp. Cryptococcus sp. was identified with mycological methods. This isolation related to a young man that has a history for long contact to pigeon.some of the fungi specially yeast can be a part of mycoflora in digestive system but the finding of Cryptococcus is uncommon. Conclusion: In this study the fungi were isolated from 5% of appendisits and with pay attention to this finding that the most patients hadn.t background factors causing the proliferation of the fungal agents in the intestine, so with further studies it is probable to consider the fungi as the agents causing appendicitis in this patients.

  19. In vitro attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia homoeocarpa.

    Science.gov (United States)

    Allen, Tom W; Burpee, Leon L; Buck, James W

    2004-12-01

    The ability of yeasts to attach to hyphae or conidia of phytopathogenic fungi has been speculated to contribute to biocontrol activity on plant surfaces. Attachment of phylloplane yeasts to Botrytis cinerea, Rhizoctonia solani, and Sclerotinia homoeocarpa was determined using in vitro attachment assays. Yeasts were incubated for 2 d on potato dextrose agar (PDA) prior to experimentation. A total of 292 yeasts cultured on PDA were screened for their ability to attach to conidia of B. cinerea; 260 isolates (89.1%) attached to conidia forming large aggregates of cells, and 22 isolates (7.5%) weakly attached to conidia with 1 or 2 yeast cells attached to a few conidia. Ten yeasts (3.4%), including 8 isolates of Cryptococcus laurentii, 1 isolate of Cryptococcus flavescens, and an unidentified species of Cryptococcus, failed to attach to conidia. All non-attaching yeasts produced copious extracellular polysaccharide (EPS) on PDA. Seventeen yeast isolates did not attach to hyphal fragments of B. cinerea, R. solani, and S. homoeocarpa after a 1 h incubation, but attachment was observed after 24 h. Culture medium, but not culture age, significantly affected the attachment of yeast cells to conidia of B. cinerea. The 10 yeast isolates that did not attach to conidia when grown on agar did attach to conidia (20%-57% of conidia with attached yeast cells) when cultured in liquid medium. Attachment of the biocontrol yeast Rhodotorula glutinis PM4 to conidia of B. cinerea was significantly greater at 1 x 10(7) yeast cells x mL(-1) than at lower concentrations of yeast cells. The ability of yeast cells to attach to fungal conidia or hyphae appears to be a common phenotype among phylloplane yeasts.

  20. Relationship between intracranial pressure and antifungal agents levels in the CSF of patients with cryptococcal meningitis.

    Science.gov (United States)

    Wirth, Fernanda; de Azevedo, Maria Isabel; Pilla, Carmen; Aquino, Valério Rodrigues; Neto, Gustavo Wissmann; Goldani, Luciano Zubaran

    2018-04-01

    The purpose of this study was to evaluate the influence of intracranial hypertension in the cerebrospinal fluid (CSF) levels of amphotericin B and fluconazole levels of patients with cryptococcal meningitis. CSF samples and intracranial pressure were obtained by means of routine punctures performed at days 1, 7, and 14 of therapy, respectively. Amphotericin B and fluconazole CSF levels were measured by HPLC method as previously described. The minimum inhibitory concentration for amphotericin B, fluconazole, 5΄flucytosine, and voriconazole of each Cryptococcus isolate was performed according to CLSI. The predominant Cryptococcus species found was C. neoformans, and the major underlying condition was AIDS. Only one CSF sample had a detectable level for amphotericin B during the 14 days of therapy. Fluconazole CSF levels progressively increased from day 1 to day 14 of therapy for most cases. Fluconazole levels in the CSF were above the minimum inhibitory concentrations (MICs) for Cryptococcus during the initial 14 days of antifungal therapy. Variations of intracranial pressure did not affect amphotericin B and fluconazole levels in the CSF. The generalized estimating correlation (GEE) and Spearman correlation test (SCT) showed no significant correlation between the amphotericin B or fluconazole concentrations in the CSF and intracranial pressure (P = .953 and P = .093, respectively for GEE test and P = .477 and P = .847, respectively, for SCT). Combination therapy of amphotericin B with fluconazole was effective in 60% of the patients considering CSF cultures were negative in 9 of 15 patients after 14 days of therapy. Further studies are necessary to evaluate the role of intracranial hypertension on the therapeutic efficacy of different antifungal agents in patients with cryptococcal meningitis.

  1. The diversity, extracellular enzymatic activities and photoprotective compounds of yeasts isolated in Antarctica

    Directory of Open Access Journals (Sweden)

    Aline B. M Vaz

    2011-09-01

    Full Text Available The diversity of yeasts collected from different sites in Antarctica (Admiralty Bay, King George Island and Port Foster Bay and Deception Island and their ability to produce extracellular enzymes and mycosporines were studied. Samples were collected during the austral summer season, between November 2006 and January 2007, from the rhizosphere of Deschampsia antarctica, ornithogenic (penguin guano soil, soil, marine and lake sediments, marine water and freshwater from lakes. A total of 89 isolates belonging to the following genera were recovered: Bensingtonia, Candida, Cryptococcus, Debaryomyces, Dioszegia, Exophiala, Filobasidium, Issatchenkia (Pichia, Kodamaea, Leucosporidium, Leucosporidiella, Metschnikowia, Nadsonia, Pichia, Rhodotorula, and Sporidiobolus, and the yeast-like fungi Aureobasidium, Leuconeurospora and Microglossum. Cryptococcus victoriae was the most frequently identified species. Several species isolated in our study have been previously reported to be Antarctic psychophilic yeasts, including Cr. antarcticus, Cr. victoriae, Dioszegia hungarica and Leucosporidium scottii. The cosmopolitan yeast species A. pullulans, C. zeylanoides, D. hansenii, I. orientalis, K. ohmeri, P. guilliermondii, Rh. mucilaginosa, and S. salmonicolor were also isolated. Five possible new species were identified. Sixty percent of the yeasts had at least one detectable extracellular enzymatic activity. Cryptococcus antarcticus, D. aurantiaca, D. crocea, D. hungarica, Dioszegia sp., E. xenobiotica, Rh. glaciales, Rh. laryngis, Microglossum sp. 1 and Microglossum sp. 2 produced mycosporines. Of the yeast isolates, 41.7% produced pigments and/or mycosporines and could be considered adapted to survive in Antarctica. Most of the yeasts had extracellular enzymatic activities at 4ºC and 20ºC, indicating that they could be metabolically active in the sampled substrates.

  2. Interaction between ketoconazole, amphotericin B and terbinafin and three diazenumdiolates in concomitant uses against some fugal species

    Directory of Open Access Journals (Sweden)

    Mehraban Falahati

    2006-05-01

    Full Text Available A checkerboard broth microdilution method was performed to investigate the in vitro antifungal activities of three diazeniumdiolates derivatives (DETA/NO, DPTA/NO, DEA/NO alone and in combination with ketoconazole, amphotricin B or terbinafine against five Candida species, Cryptococcus neoformance and four dermatophyte strains. MICs and MLCs were recorded, and synergy was calculated by using fractional inhibitory and fractional lethal concentration index. DETA/NO with a half-life of 57h at 25°C showed antifungal activity against all tested dermatophyte species (MIC 0.150 to 2.5mg/ml, DPTA/NO with a half life of 3h at 37°C showed antifungal activity against five species of Candida and Cryptococcus neoformans, and DEA/NO with a half life of 2 min at 37°C and 16 min at 25°C did not show antifungal activity against tested strains. Combinations of DPTA-NO with either ketoconazole or amphotericin B were either synergistic or indifferent for all tested strain of Candida and Cryptococcus neoformance. DETA/NO was unable to enhance the antifungal activity of terbinafine against dermatophyte strains. Even where no synergistic activity was achieved, there was still a decrease in the MIC of one or both drugs which were used in combination. Antagonism was observed between terbinafine and DETA-NO against Trichophyton rubrum. Our result suggests that DETA/NO and DPTA/NO may be useful for development of new therapeutic strategies for treatment of dermatophyte and Candida infections. Clinical studies are warranted to elucidate the potential utility of these combination therapies.

  3. Positive selection for unpreferred codon usage in eukaryotic genomes

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    Galagan James E

    2007-07-01

    Full Text Available Abstract Background Natural selection has traditionally been understood as a force responsible for pushing genes to states of higher translational efficiency, whereas lower translational efficiency has been explained by neutral mutation and genetic drift. We looked for evidence of directional selection resulting in increased unpreferred codon usage (and presumably reduced translational efficiency in three divergent clusters of eukaryotic genomes using a simple optimal-codon-based metric (Kp/Ku. Results Here we show that for some genes natural selection is indeed responsible for causing accelerated unpreferred codon substitution, and document the scope of this selection. In Cryptococcus and to a lesser extent Drosophila, we find many genes showing a statistically significant signal of selection for unpreferred codon usage in one or more lineages. We did not find evidence for this type of selection in Saccharomyces. The signal of positive selection observed from unpreferred synonymous codon substitutions is coincident in Cryptococcus and Drosophila with the distribution of upstream open reading frames (uORFs, another genic feature known to reduce translational efficiency. Functional enrichment analysis of genes exhibiting low Kp/Ku ratios reveals that genes in regulatory roles are particularly subject to this type of selection. Conclusion Through genome-wide scans, we find recent selection for unpreferred codon usage at approximately 1% of genetic loci in a Cryptococcus and several genes in Drosophila. Unpreferred codons can impede translation efficiency, and we find that genes with translation-impeding uORFs are enriched for this selection signal. We find that regulatory genes are particularly likely to be subject to selection for unpreferred codon usage. Given that expression noise can propagate through regulatory cascades, and that low translational efficiency can reduce expression noise, this finding supports the hypothesis that translational

  4. Yeast species associated with wine grapes in China.

    Science.gov (United States)

    Li, Shuang-Shi; Cheng, Chao; Li, Zheng; Chen, Jing-Yu; Yan, Bin; Han, Bei-Zhong; Reeves, Malcolm

    2010-03-31

    Having more information on the yeast ecology of grapes is important for wine-makers to produce wine with high quality and typical attributes. China is a significant wine-consuming country and is becoming a serious wine-producer, but little has been reported about the yeast ecology of local ecosystems. This study provides the first step towards the exploitation of the yeast wealth in China's vine-growing regions. The aim of this study was to investigate the yeast population density and diversity on three grape varieties cultivated in four representative vine-growing regions of China. Yeast species diversity was evaluated by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequence analysis of the 5.8S internal transcribed spacer (ITS) ribosomal DNA (rDNA) region of cultivable yeasts. The grapes harbored yeast populations at 10(2)-10(6)CFU/mL, consisting mostly of non-Saccharomyces species. Seventeen different yeast species belonging to eight genera were detected on the grape samples tested, including Hanseniaspora uvarum, Cryptococcus flavescens, Pichia fermentans, Candida zemplinina, Cryptococcus carnescens, Candida inconpicua, Zygosaccharomyces fermentati, Issatchenkia terricola, Candida quercitrusa, Hanseniaspora guilliermondii, Candida bombi, Zygosaccharomyces bailii, Sporidiobolus pararoseus, Cryptococcus magnus, Metschnikowia pulcherrima, Issatchenkia orientalis and Pichia guilliermondii. H. uvarum and C. flavescens were the dominant species present on the grapes. For the first time Sporidiobolus pararoseus was discovered as an inhabitant of the grape ecosystem. The yeast community on grape berries was influenced by the grape chemical composition, vine-variety and vine-growing region. This study is the first to identify the yeast communities associated with grapes in China using molecular methods. The results enrich our knowledge of wine-related microorganisms, and can be used to promote the development of the local wine

  5. Chemical composition, toxicity and larvicidal and antifungal activities of Persea americana (avocado) seed extracts.

    Science.gov (United States)

    Leite, João Jaime Giffoni; Brito, Erika Helena Salles; Cordeiro, Rossana Aguiar; Brilhante, Raimunda Sâmia Nogueira; Sidrim, José Júlio Costa; Bertini, Luciana Medeiros; Morais, Selene Maia de; Rocha, Marcos Fábio Gadelha

    2009-01-01

    The present study had the aim of testing the hexane and methanol extracts of avocado seeds, in order to determine their toxicity towards Artemia salina, evaluate their larvicidal activity towards Aedes aegypti and investigate their in vitro antifungal potential against strains of Candida spp, Cryptococcus neoformans and Malassezia pachydermatis through the microdilution technique. In toxicity tests on Artemia salina, the hexane and methanol extracts from avocado seeds showed LC50 values of 2.37 and 24.13 mg mL-1 respectively. Against Aedes aegypti larvae, the LC50 results obtained were 16.7 mg mL-1 for hexane extract and 8.87 mg mL-1 for methanol extract from avocado seeds. The extracts tested were also active against all the yeast strains tested in vitro, with differing results such that the minimum inhibitory concentration of the hexane extract ranged from 0.625 to 1.25mg L-(1), from 0.312 to 0.625 mg mL-1 and from 0.031 to 0.625 mg mL-1, for the strains of Candida spp, Cryptococcus neoformans and Malassezia pachydermatis, respectively. The minimal inhibitory concentration for the methanol extract ranged from 0.125 to 0.625 mg mL-1, from 0.08 to 0.156 mg mL-1 and from 0.312 to 0.625 mg mL-1, for the strains of Candida spp., Cryptococcus neoformans and Malassezia pachydermatis, respectively.

  6. Cryptococcal breast abscess in an HIV-positive patient: arguments for reviewing the definition of immune reconstitution inflammatory syndrome.

    Science.gov (United States)

    Haddow, Lewis J; Sahid, Faieza; Moosa, Mahomed-Yunus S

    2008-07-01

    Atypical manifestations of Cryptococcus neoformans disease have been reported in patients with HIV-1 infection as part of the spectrum of the immune reconstitution inflammatory syndrome (IRIS). We describe a cryptococcal breast abscess in a patient presenting after 11 months of highly active antiretroviral therapy (HAART). The arguments for and against the case being a novel manifestation of IRIS are discussed. The potential hazards of using CD4 count as a surrogate marker of IRIS and the danger of misdiagnosing IRIS as failure of HAART are highlighted.

  7. Antifungal saponins from Swartzia langsdorffii; Saponinas antifungicas de Swartzia langsdorffii

    Energy Technology Data Exchange (ETDEWEB)

    Marqui, Sara Regina de; Lemos, Renata Brionizio; Santos, Luciana Avila; Castro-Gamboa, Ian; Cavalheiro, Alberto Jose; Bolzani, Vanderlan da Silva; Silva, Dulce Helena Siqueira [Universidade Estadual Paulista (UNESP), Araraquara, SP (Brazil). Inst. de Quimica]. E-mail: dhsilva@iq.unesp.br; Scorzoni, Liliana; Fusco-Almeida, Ana Maria; Mendes-Giannini, Maria Jose Soares [Universidade Estadual Paulista (UNESP), Araraquara, SP (Brazil). Faculdade de Ciencias Farmaceuticas; Young, Maria Claudia Marx; Torres, Luce Maria Brandao [Inst. de Botanica, SP (Brazil). Secao de Fisiologia e Bioquimica de Plantas

    2008-07-01

    Chromatographic fractionation of the EtOH extract from the leaves of Swartzia langsdorffii afforded the pentacyclic triterpenes oleanolic acid and lupeol, and two saponins: oleanolic acid 3-sophoroside and the new ester 3-O-{beta}-D-(6'-methyl)-glucopyranosyl-28-O-{beta}-D-glucopyranosyl-oleanate. Their structures were elucidated from spectral data, including 2D NMR and HRESIMS experiments. Antifungal activity of all isolated compounds was evaluated, using phytopathogens Cladosporium cladosporioides and C. sphaerospermum, and human pathogens Candida albicans, C. krusei, C. parapsilosis and Cryptococcus neoformans. (author)

  8. Antifungal Activity of Decyl Gallate against Several Species of Pathogenic Fungi

    Directory of Open Access Journals (Sweden)

    Ana Carolina Alves de Paula e Silva

    2014-01-01

    Full Text Available This work aims to demonstrate that the gallic acid structure modification to the decyl gallate (G14 compound contributed to increase the antifungal activity against several species of pathogenic fungi, mainly, Candida spp., Cryptococcus spp., Paracoccidioides spp., and Histoplasma capsulatum, according to standardized microdilution method described by Clinical Laboratory Standard Institute (CLSI documents. Moreover this compound has a particularly good selectivity index value, which makes it an excellent candidate for broad-spectrum antifungal prototype and encourages the continuation of subsequent studies for the discovery of its mechanism of action.

  9. Aurantoside K, a New Antifungal Tetramic Acid Glycoside from a Fijian Marine Sponge of the Genus Melophlus

    Directory of Open Access Journals (Sweden)

    Rohitesh Kumar

    2012-01-01

    Full Text Available A new tetramic acid glycoside, aurantoside K, was isolated from a marine sponge belonging to the genus Melophlus. The structure of the compound was elucidated on the basis of spectroscopic analysis (1H NMR, 1H–1H COSY, HSQC, and HMBC, as well as high-resolution ESILCMS. Aurantoside K did not show any significant activity in antimalarial, antibacterial, or HCT-116 cytotoxicity assays, but exhibited a wide spectrum of antifungal activity against wild type Candida albicans, amphotericin-resistant C. albicans, Cryptococcus neoformans, Aspergillus niger, Penicillium sp., Rhizopus sporangia and Sordaria sp.

  10. Effects of Food Based Yeast on Oxidant-Antioxidant Systems in Rats fed by High Cholesterol Diet

    OpenAIRE

    Savaş, Hasan Basri; Yüksel, Özlem; Şanlıdere Aloğlu, Hatice; Öner, Zübeyde; Demir Özer, Ezgi; Gültekin, Fatih

    2013-01-01

    In living organisms, oxidant and antioxidant systems are in a balance. In the present study, our aim was to study the effects of Cryptococcus humicola, which is a food based yeast whose cholesterol lowering activity is under investigation, on oxidant and antioxidant systems.31 adult male, Wistar albino rats weighing 200-250 gr were included in the study. Rats were divided into four groups based on their diets. Group 1(Control Group) was fed a normal diet, Group 2 was fed a high cholesterol di...

  11. Antifungal saponins from Swartzia langsdorffii

    International Nuclear Information System (INIS)

    Marqui, Sara Regina de; Lemos, Renata Brionizio; Santos, Luciana Avila; Castro-Gamboa, Ian; Cavalheiro, Alberto Jose; Bolzani, Vanderlan da Silva; Silva, Dulce Helena Siqueira; Scorzoni, Liliana; Fusco-Almeida, Ana Maria; Mendes-Giannini, Maria Jose Soares; Young, Maria Claudia Marx; Torres, Luce Maria Brandao

    2008-01-01

    Chromatographic fractionation of the EtOH extract from the leaves of Swartzia langsdorffii afforded the pentacyclic triterpenes oleanolic acid and lupeol, and two saponins: oleanolic acid 3-sophoroside and the new ester 3-O-β-D-(6'-methyl)-glucopyranosyl-28-O-β-D-glucopyranosyl-oleanate.Their structures were elucidated from spectral data, including 2D NMR and HRESIMS experiments. Antifungal activity of all isolated compounds was evaluated, using phytopathogens Cladosporium cladosporioides and C. sphaerospermum, and human pathogens Candida albicans, C. krusei, C. parapsilosis and Cryptococcus neoformans. (author)

  12. Antimicrobial and antiviral activities of polyphenolics from Cocos nucifera Linn. (Palmae) husk fiber extract.

    Science.gov (United States)

    Esquenazi, Daniele; Wigg, Marcia D; Miranda, Mônica M F S; Rodrigues, Hugo M; Tostes, João B F; Rozental, Sonia; da Silva, Antonio J R; Alviano, Celuta S

    2002-12-01

    The decoction of Cocos nucifera L. husk fiber has been used in northeastern Brazil traditional medicine for treatment of diarrhea and arthritis. Water extract obtained from coconut husk fiber and fractions from adsorption chromatography revealed antimicrobial activity against Staphylococcus aureus. The crude extract and one of the fractions rich in catechin also showed inhibitory activity against acyclovir-resistant herpes simplex virus type 1 (HSV-1-ACVr). All fractions were inactive against the fungi Candida albicans, Fonsecaea pedrosoi and Cryptococcus neoformans. Catechin and epicatechin together with condensed tannins (B-type procyanidins) were demonstrated to be the components of the water extract.

  13. Presentación sistémica de Aspergillus spp con semiología neurológica en un Pastor Alemán: informe de un caso clínico

    OpenAIRE

    Hernández Tapia, Ana Laura; Santoscoy Mejía, Eduardo Carlos; Ramírez Lezama, José; Cervantes Olivares, Roberto A.; Quiroz Mercado, Joaquín Arturo

    2010-01-01

    La infección del sistema nervioso en el perro es poco común; posibles causas son Aspergillus spp, Cryptococcus neoformans, Blastomyces dermatidis y Coccidioides immitis. La infección por Aspergillus spp afecta al aparato respiratorio y en pocas ocasiones se presenta de manera diseminada. Sin embargo, en el Pastor Alemán hay mayor incidencia de éste, pues dicha infección se asocia con una deficiencia inmunológica. Se describe el caso de una perra de raza Pastor Alemán, de 1.5 años de edad, ref...

  14. Disseminated cryptococcosis with splenic and meningeal involvement in a patient with AIDS

    International Nuclear Information System (INIS)

    Lizarazo, Jairo; Parra, Edgar; Parada, Oscar

    2010-01-01

    We present the case of a 37-year-old woman with a recent diagnosis of HIV-infection. The patient consulted because of headache, diarrhea, vomit and fever of two weeks' evolution. The patient had a wasting syndrome, and severe splenomegaly. Cerebral spinal fluid study showed meningitis by Cryptococcus neoformans. CT-scan revealed splenomegaly with focal lesions. Percutaneous biopsy with 18-gauge needle from one of the focal lesions confirmed cryptococcosis. The treatment was initiated with amphotericin B, followed by fluconazole. The patient's evolution has been satisfactory. Splenic lesions resolved and the patient is now asymptomatic and on secondary prophylaxis with fluconazole and antiretroviral treatment

  15. Comparison of BacT/Alert FAN and FAN Plus Bottles with Conventional Medium for Culturing Cerebrospinal Fluid.

    Science.gov (United States)

    Yoo, In Young; Chun, Sejong; Song, Dong Joon; Huh, Hee Jae; Lee, Nam Yong

    2016-11-01

    We compared the BacT/Alert system FAN and FAN Plus media to conventional media for culturing cerebrospinal fluid (CSF) with 2,545 samples. FAN/FAN Plus bottles showed better performance for isolating microorganisms in CSF than conventional media (positive rate, 7.2% [182/2,545] versus 3.1% [80/2,545]). The incremental recovery rate of Cryptococcus neoformans from FAN Plus bottles was higher than that from FAN bottles. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  16. Central Nervous System Strongyloidiasis and Cryptococcosis in an HIV-Infected Patient Starting Antiretroviral Therapy

    Directory of Open Access Journals (Sweden)

    Mónica Rodríguez

    2012-01-01

    Full Text Available We report a case of Strongyloides stercoralis hyperinfection syndrome with central nervous system involvement, in a patient with late human immunodeficiency virus (HIV infection starting antiretroviral therapy, in whom Strongyloides stercoralis larvae and Cryptococcus neoformans were isolated antemortem from cerebrospinal fluid. Our patient was not from an endemic region for the parasite, so strongyloidiasis was not originally suspected. For this reason, we conclude that Strongyloides stercoralis infection should be suspected in HIV-infected patients starting antiretroviral therapy in order to avoid potential fatal outcomes.

  17. Sesquiterpene-derived metabolites from the deep water marine sponge Poecillastra sollasi.

    Science.gov (United States)

    Killday, K B; Longley, R; McCarthy, P J; Pomponi, S A; Wright, A E; Neale, R F; Sills, M A

    1993-04-01

    Six sesquiterpene-derived compounds, 1-6, which we call sollasins a-f, have been isolated from a deep water specimen of the sponge Poecillastra sollasi. The structures were elucidated by comparison of spectral data to related metabolites and confirmed using spectroscopic methods. The compounds inhibit the growth of the pathogenic fungi Candida albicans and Cryptococcus neoformans and the P-388 and A-549 tumor cell lines. Compounds 3 and 4 show weak inhibition of binding of [125I] angiotensin II to rat aorta smooth muscle cell membranes.

  18. Industrially Important Carbohydrate Degrading Enzymes from Yeasts: Pectinases, Chitinases, and β-1,3-Glucanases

    Science.gov (United States)

    Gummadi, Sathyanarayana N.; Kumar, D. Sunil; Dash, Swati S.; Sahu, Santosh Kumar

    Polysaccharide degrading enzymes are hydrolytic enzymes, which have a lot of industrial potential and also play a crucial role in carbon recycling. Pectinases, chitinases and glucanases are the three major polysaccharide degrading enzymes found abundantly in nature and these enzymes are mainly produced by fungal strains. Production of these enzymes by yeasts is advantageous over fungi, because the former are easily amenable to genetic manipulations and time required for growth and production is less than that of the latter. Several yeasts belonging to Saccharomyces, Pichia, Rhodotorula and Cryptococcus produce extracellular pectinases, glucanases and chitinases. This chapter emphasizes on the biological significance of these enzymes, their production and their industrial applications.

  19. Epidemiologic study of bronchopulmonary mycosis in the province of cordoba, argentina

    Directory of Open Access Journals (Sweden)

    Diana T. Masih

    1987-02-01

    Full Text Available An epidemiological study for histoplasmosis coccidioidomycosis and cryptococcosis made in five areas of the province of Córdoba is presented. The data obtained showed a global positivity of 41.1% for histoplasmin 26.7% for coccidioidin and 14.1% for cryptococcin. In some areas, the Rio III basin and Traslasierra, the histoplasmosis infection indexes were much higher, 53,3% and 73.1% respectively. The index of positive skin tests with Cryptococcus antigen in Traslasierra was also very high: 31.9%.

  20. Yeasts in pigeon feacal droppings in Lisbon - Portugal, 1994

    Directory of Open Access Journals (Sweden)

    Herminia Maria Lourdes Martins

    1997-10-01

    Full Text Available In this work, the results of a preliminary survey held in city of Lisbon. Eighty faecal samples were examined between Summer and Autumn, 1994, from twelve different urban areas, mainly near churchs and monuments where birds nest, rest or eat. From each sample 1 g was weighted and suspensed in 10 ml of destilled sterilized water and consecutive decimal diluitions were executed. Yeasts were enumerated and grouped by species, based on morphological types. On eighty faecal samples the most prevalent yeasts identified were: Candida humicola (51.5%, Candida albicans (48.7%, Cryptococcus neoformans (5% and Trichosporon cutaneum (37.5%.