• Title/Summary/Keyword: Cryptococcus

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Delineating Transcription Factor Networks Governing Virulence of a Global Human Meningitis Fungal Pathogen, Cryptococcus neoformans

  • Jung, Kwang-Woo;Yang, Dong-Hoon;Maeng, Shinae;Lee, Kyung-Tae;So, Yee-Seul;Hong, Joohyeon;Choi, Jaeyoung;Byun, Hyo-Jeong;Kim, Hyelim;Bang, Soohyun;Song, Min-Hee;Lee, Jang-Won;Kim, Min Su;Kim, Seo-Young;Ji, Je-Hyun;Park, Goun;Kwon, Hyojeong;Cha, Sooyeon;Meyers, Gena Lee;Wang, Li Li;Jang, Jooyoung;Janbon, Guilhem;Adedoyin, Gloria;Kim, Taeyup;Averette, Anna K.;Heitman, Joseph;Cheong, Eunji;Lee, Yong-Hwan;Lee, Yin-Won;Bahn, Yong-Sun
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.59-59
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    • 2015
  • Cryptococcus neoformans causes life-threatening meningoencephalitis in humans, but the treatment of cryptococcosis remains challenging. To develop novel therapeutic targets and approaches, signaling cascades controlling pathogenicity of C. neoformans have been extensively studied but the underlying biological regulatory circuits remain elusive, particularly due to the presence of an evolutionarily divergent set of transcription factors (TFs) in this basidiomycetous fungus. In this study, we constructed a high-quality of 322 signature-tagged gene deletion strains for 155 putative TF genes, which were previously predicted using the DNA-binding domain TF database (http://www.transcriptionfactor.org/). We tested in vivo and in vitro phenotypic traits under 32 distinct growth conditions using 322 TF gene deletion strains. At least one phenotypic trait was exhibited by 145 out of 155 TF mutants (93%) and approximately 85% of the TFs (132/155) have been functionally characterized for the first time in this study. Through high-coverage phenome analysis, we discovered myriad novel TFs that play critical roles in growth, differentiation, virulence-factor (melanin, capsule, and urease) formation, stress responses, antifungal drug resistance, and virulence. Large-scale virulence and infectivity assays in insect (Galleria mellonella) and mouse host models identified 34 novel TFs that are critical for pathogenicity. The genotypic and phenotypic data for each TF are available in the C. neoformans TF phenome database (http://tf.cryptococcus.org). In conclusion, our phenome-based functional analysis of the C. neoformans TF mutant library provides key insights into transcriptional networks of basidiomycetous fungi and ubiquitous human fungal pathogens.

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Microbe Hunting: A Curious Case of Cryptococcus

  • Bartlett, Karen H.;Kidd, Sarah;Duncan, Colleen;Chow, Yat;Bach, Paxton;Mak, Sunny;MacDougall, Laura;Fyfe, Murray
    • Proceedings of the Korean Environmental Health Society Conference
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    • 2005.06a
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    • pp.45-72
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    • 2005
  • C. neoformans-associated cryptococcosis is primarily a disease of immunocompromised persons, has a world-wide distribution, and is often spread by pigeons in the urban environment. In contrast, C. gattii causes infection in normal hosts, has only been described in tropical and semi-tropical areas of the world, and has a unique niche in river gum Eucalyptus trees. Cryptococcosis is acquired through inhalation of the yeast propagules from the environment. C. gattii has been identified as the cause of an emerging infectious disease centered on Vancouver Island, British Columbia, Canada. No cases of C. gattii-disease were diagnosed prior to 1999; the current incidence rate is 36 cases per million population. A search was initiated in 2001 to find the ecological niche of this basidiomycetous yeast. C. gaftii was found in the environment in treed areas of Vancouver Island. The highest percentage of colonized-tree clusters were found around central Vancouver Island, with decreasing rates of colonization to the north and south. Climate, soil and vegetation cover of this area, called the Coastal Douglas fir biogeoclimatic zone, is unique to British Columbia and Canada. The concentration of airborne C. gattii was highest in the dry summer months, and lowest during late fall, winter, and early spring, months which have heavy rainfall. The study of the emerging colonization of this organism and subsequent cases of environmentally acquired disease will be informative in planning public health management of new routes of exposure to exotic agents in areas impacted by changing climate and land use patterns. Cryptococcosis is an infection associated with an encapsulated, basidiomycetous yeast Cryptococcus neoformans. The route of entry for this organism is through the lungs, with possible systemic spread via the circulatory system to the brain and meninges. There are four cryptococcal serogroups associated with disease in humans and animals, distinguished by capsular polysaccharide antigens. Cryptococcus neoformans: variety grubii (serotype A), variety neoformans (serotype D), and variety gattii (serotypes B and C) (Franzot et at. 1999). C. neoformans variety gattii has recently been elevated to species status, C. gattii. C. neoformans val. grubii and var. neoformans have a world-wide distribution, and are particularly associated with soil and weathered bird droppings. In contrast, C. gattii (CG) is not associated with bird excrement, is primarily found in tropical and subtropical climates, and has a restricted environmental niche associated with specific tree species. (Ellis & Pfiffer 1990) Ellis and Pfeiffer theorize that, as a basidiomycete, CG requires an association with a tree in order to become pathogenic to mammals. In Australia, CG has been found to be associated with five species of Eucalypts, Eucalyptus camaldulensis, E. tereticornis, E. blakelyi, E. gomphocephala, and E. rudis. Eucalypts, although originally native to Australia, now have a world-wide distribution. CG has been found associated with imported eucalypts in India, California, Brazil, and Egypt. In addition, in Brazil and Columbia, where eucalypts have been naturalized, native trees have been shown to harbour CG (Callejas et al. 1998; Montenegro et al. 2000). In British Columbia, Canada, since the beginning of 1999, there have been 120 confirmed cases of cryptococcal mycoses associated with CG in humans, including 4 fatalities (data from British Columbia Centre for Disease Control), and over 200 cases in animal pets in BC (data from Central Laboratory for Veterinarians). What is remarkable about the BC outbreak of C. gattii-cryptococcosis is that all of the cases have been residents of, or visitors to, a narrow area along the eastern coast of Vancouver Island, BC, from the tip of the island in the south (Victoria) to Courtenay on the north-central island as illustrated in Figure 1. Of the first 38 human cases, 58% were male with a mean age of 59.7 years (range 20 - 82): 36 cases (95%) were Caucasian. Ten cases (26%) presented with meningitis, the remainder presented with respiratory symptoms. Cultures recovered from cases of cryptococcosis associated with the outbreak were typed as serogroup B, which is specific to CG (Bartlett et al. 2003). This was the first reported outbreak of CVG in Canada, or indeed, the world. Where infection with CG is endemic, for example, Australia, the incidence of cryptococcosis ranges from 1.8 - 4.7 per million between the southern and northern states (Sorrell 2001). However, the overall incidence of cryptococcosis in immunocompenent individuals has been estimated at 0.2 per million population per year (Kwon-Chung et al. 1984). The population of Vancouver Island is approximately 720,000,consequently, even if the organism were endemic, one would expect a maximum of 0.15 cases of cryptococcal disease annually.

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Isolation of Wild Yeasts from the Water and Riverside Soil of Geumgang Midstream in Sejong City, Korea, and Characterization of Unrecorded Wild Yeasts (세종특별자치시 주변의 금강 중류 물과 토양에서 야생 효모의 분리 및 국내 미기록 효모의 특성)

  • Han, Sang-Min;Kim, Ji-Yoon;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.51-61
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    • 2019
  • The goal of this study was to elucidate wild yeast diversity of Geumgang midstream near Sejong metropolitan autonomous city, Korea. Thirty-seven strains of 32 species of wild yeasts were isolated from 43 water and soil samples under the Bulti bridge of Sejong city, Korea. Seven yeasts of each Candida spp. and Cryptococcus spp. were the predominant species isolated from samples near the Bulti bridge. Holtermanniella takashimae SW048 (NNIBRFG9314), Cystofilobasidium infirmominiatum SW013 (NNIBRFG9310), Mrakia cryoconite SW015 (NNIBRFG9316), Pichia sporocuriosa SW085 (NNIBRFG9326) and Cryptococcus aspenensis SW008 (NNIBRFG9309) represented novel yeast strains found in Korea for the first time. All of these previously unrecorded yeasts, except for Mrakia cryoconite SW015 had ascospores and grew well in yeast extract-peptone-dextrose (YPD), yeast extract-malt extract (YM) and potato-extrose (PD) media. Pichia sporocuriosa SW085 grew well in vitamin-free medium and Holtermanniella takashimae SW048, which was a halotolerant wild yeast, grew well YPD medium containing 5 % NaCl. Twenty-six strains representing eight species of wild yeast were isolated from 22 water and soil samples under the Haetmuri bridge of Sejong city, Korea. Candida pseudolambica (12 strains) and Aureobasidium pullulans (11 strains) were the predominant isolates from samples near the Haetmuri bridge. Occultifur kilbournensis HB060 (NNIBRFG9317), Sampaiozyma vanillica HB014 (NNIBRFG9332), Xenoramularia neerlandica HB039 (NNIBRFG9335), Candida norvegica HB315 (NNIBRFG9306), C. melibiosica HB316 (NNIBRFG9305), C. quercuum GB014 (NNIBRFG9307), and C. succiphila GB015 (NNIBRFG9308) represented novel yeast strains recorded in Korea for the first time. O. kilbournensis HB060 and X. neerlandica HB039 did not form ascospores or pseudo-mycelia. All of these previously unrecorded yeasts, except S. vanillica HB014 and X. neerlandica HB039, grew well in vitaminfree medium, and C. norvegica HB315 and C. succiphila GB015, which were halotolerant wild yeasts, which grew well in YPD medium containing 5 % NaCl.

Phylogeny of the Yeast Species Isolated from Wild Tiger Lily (Lilium lancifolium Thunb.) (야생 참나리(Lilium lancifolium Thunb.)로부터 분리한 효모의 분자계통학적 분석)

  • Kim, Jong-Shik;Kim, Dae-Shin
    • Korean Journal of Environmental Agriculture
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    • v.34 no.2
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    • pp.149-154
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    • 2015
  • BACKGROUND: Yeast isolates associated with the leaves, stems, and flowers of the tiger lily needed to be identified using isolation methods that have previously been used effectively in yeast biotechnology. A culture-based approach was necessary for the isolation of many yeast strains associated with tiger lily. METHODS AND RESULTS: In this study, the homogenized leaves, stems, and flowers of tiger lily were spreaded onto GPY medium containing chloramphenicol, streptomycin, Triton X-100, and L-sorbose. A total of 82 yeast strains from the leaves, 94 and 97 yeast strains from the stems and flowers were isolated, respectively. Yeast isolates were identified by phylogenetic analysis based on internal transcribed spacer region sequencing. The yeast species isolated from the leaves comprised of 31 isolates of the genus Pseudozyma, 28 of Aureobasidium pullulans, and 11 of the genus Cryptococcus. Those isolated from the stems comprised of 40 of A. pullulans and 11 of Cryptococcus, and 95 of A. pullulans While, 1 isolate each of the genera Rhodotorula and Metschnikowia were isolated from the flowers. CONCLUSION: We identified site-specific yeast communities associated with tiger lily. These yeast isolates may have high potential for application in the field of biotechnology.

Genetic Diversity of Culturable Endophytic Fungi Isolated from Halophytes Naturally Growing in Muan Salt Marsh (무안 염습지에 자생하는 염생식물(halophyte)의 뿌리로부터 분리된 내생진균(endophytic fungi)의 유전학적 다양성)

  • You, Young-Hyun;Yoon, Hyeok-Jun;Seo, Yeong-Gyo;Kim, Mi-Ae;Kang, Myung-Suk;Kim, Chang-Mu;Ha, Sang-Chul;Cho, Ga-Youn;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.7
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    • pp.970-980
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    • 2012
  • Native halophytes, such as Suaeda maritima, Limonium tetragonum, S. japonica, Zoysia sinica, and Phragmites australis were collected from the Muan salt marsh. Ninety endophytic fungi were isolated from the roots of the collected halophytes. Molecular insights inferred by internal transcribed spacer containing ITS1, 5.8s, and the ITS2 region showed that all the fungal strains belong to ten orders, i.e., Capnodiales (4.44%), Cystofilobasidiales (1.11%), Dothideales (3.33%), Eurotiales (53.33%), Glomerellales (3.33%), Hypocreales (8.89%), Mucorales (1.11%), Pleosporales (15.56%), Sordariales (1.11%), and Trichosphaeriales (1.11%). The rest (6.67%) of all fungal isolates were not identified. Ninety fungal strains were confirmed at the genus level, containing Acremonium, Alternaria, Aspergillus, Aureobasidium, Cephalosporium, Chaetomium, Cladosporium, Colletotrichum, Cryptococcus, Didymella, Dothideomycete, Emericellopsis, Epicoccum, Eupenicillium, Fusarium, Gibberella, Gongronella, Macrophoma, Microsphaeropsis, Nigrospora, Paecilomyces, Paraconiothyrium, Penicillium, Phaeomyces, Phoma, Pleosporales, Purpureocillium, and Talaromyces. Of all the endophytic fungi identified from the various halophytes, Aspergillus and Penicillium of Eurotiales had the highest abundance.

Selective Isolation and Phylogeny of the Yeast Species Associated with Aloe vera and Aloe saponaria (알로에 베라(A. vera)와 알로에 사포나리아(A. saponaria)로 부터 효모의 분리 및 계통분석)

  • Choi, Sungchang;Kim, Myung-Uk;Kim, Jong-Shik
    • Korean Journal of Environmental Agriculture
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    • v.32 no.3
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    • pp.240-243
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    • 2013
  • BACKGROUND: Several yeast species have potential applications in biotechnology and the identification of such yeast species is of great interest. The first step in the identification of yeasts is the establishment of an effective isolation method. Thus, we compared the efficacy of different yeast media in the isolation of yeast associated with Aloe vera and Aloe saponaria. METHODS AND RESULTS: In this study, we spread homogenized A. vera and A. saponaria leaves onto 4 different yeast selective media containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose. We observed high selectivity for yeast and many colonies on media. We isolated 67 yeast strains from A. vera and 42 yeast strains from A. saponaria. We used phylogenetic analysis to identify the yeast isolates based on ITS region sequencing and performed sequence analysis on representative isolates from each agar plate. Further, we compared the sequences obtained with reference sequences. The yeast species isolated from A. vera were as follows: 56 isolates of Meyerozyma, 9 isolates of Cryptococcus, and 1 isolate each of Rhodotorula and Sporobolomyces. Those isolated from A. saponaria were as follows: 41 isolates of Rhodosporidium and 1 isolate of Sporobolomyces. CONCLUSION(S): All the isolates obtained using large agar plate containing chloramphenicol, streptomycin, Triton X-100 and L-sorbose were identified as yeast. Therefore, we concluded that this method is useful for selective screening of yeast species.

Isolation of Antifungal Substances by Bacillus amyloliquefaciens IUB158-03 and Antagonistic Activity against Pathogenic Fungi (Bacillus amyloliquefaciens IUB158-03이 생산하는 항진균물질의 분리와 항균활성)

  • Kim, Hye-Young;Lee, Tae-Soo
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.96-103
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    • 2009
  • For the control of pathogenic microorganisms, Bacillus spp. were isolated from diseased pepper fruits in Korea. Among them, Bacillus sp. IUB158-03 showed high inhibitory effect on mycelial growth and spore germination of C. gloeosporioides and Botrytis cinerea. The strain was identified as B. amyloliquefaciens IUB158-03 based on its physiological, biochemical characteristics and Microlog analysis. The highest level of antifungal substances by B. amyloliquefaciens IUB158-03 were obtained when the bacterium was cultured in medium containing 2% soluble starch, 3% yeast extract, 0.5% tryptone, 0.5% $NH_4H_2PO_4$, and 1% NaCl (pH 6.0) at $25^{\circ}C$ for 72 hrs. The antifungal substances were purified by butanol extraction, silica gel column chromatography, preparative thin layer chromatography, and high performance liquid chromatography. The purified antifungal substance was confirmed $R_f$ 0.27 by TLC. This substance exhibited antifungal activity against Fusarium solani, Rhizoctonia solani, Botrytis cineria, Alternata alternaria of plant pathogenic fungi and Trichophyton mentagrophytes, Epidermophyton floccosum, Cryptococcus neoformans of human pathogenic fungi.

An Etiologic Study of Rabbit Dermatitis at Large Rabbit Farms in South Korea (집단 사육 되는 토끼에서 호발하는 피부병에 관한 병인론적 연구)

  • Kim, Sung-Ho;Lee, Jae-Hoon;Chang, Hwa-Seok;Kang, Eun-Hee;Chung, Dai-Jung;Kim, Hwi-Yool
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1499-1505
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    • 2009
  • This study was carried out between August and September 2007 to determine the causative agents and epidemiologic features of rabbit dermatitis in Korea. Rabbits were shipped to the laboratory in the College of Veterinary Medicine from 10 rabbit farms. A total of 520 hair, blood, and skin specimens collected from skin lesions of 40 rabbits with suspected dermatopathy were examined mycologically, bacteriologically, and parasitologically. The positive rates of dermatophytosis, bacterial skin dermatitis, and ectoparasite dermatitis were 95, 92.5, and 7.5%, respectively. The etiologic agents of dermatophytosis were identified as Trichophyton mentagrophyte (95%), non-dermatophytic filamentous fungi such as Aspergillus s(5%), and Cryptococcus humilocus (2.5%). With respect to bacteria-related skin dermatitis, Staphylococcus coagulase negative was the most common etiological agent. Staphylococcus aureus was the second most frequent causative agent. Most of the pathogenic isolates were resistant to tetracycline, and aminoglycosides such as amikacin and gentamicin were the most effective drugs against the pathologic bacteria isolated. Ectoparasites were rarely detected in this study. Only Psoroptes cuniculis was detected in 3 (7.5%) out of the 40 tested rabbits. The role of ectoparasites as a causative agent of dermatitis in rabbits in this study was minimal. Our results provide important information related to rabbit dermatitis treatments and researches.

Antimicrobial and Antitumor Activities of Seed Extracts of Camellia sinensis L. (차나무 종자 추출물의 항균활성 및 항종양활성)

  • Yoon, Won-Ho;Choi, Jae-Hoon;Lee, Keyong-Ho;Kim, Chang-Han
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.108-112
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    • 2005
  • Antimicrobial and antitumor activities of Camellia sinensis L seed extracts were investigated. Seed extracts showed antifungal activities against Candida albicans IFO 1594 and Cryptococcus neoformans. Inhibition zone of 20 mm was shown by 70% ethanol extract against C. albicans IFO 1594 at 100 mg/mL. Antifungal activity of seed extract was not decreased by heating at 80 and $100^{\circ}C$ for 30 min or at $121^{\circ}C$ for 15 min, indicating heat-stability of seed component. Growth-inhibitory effects were observed in 70 and 10% of tumor cell line SK-OV-3 and normal ceil line NIH/3T3 at $50{\mu}g/mL$, respectively.

Distribution and Physiological Characteristics of Yeasts in Traditional and Commercial Kochujang (재래식 및 개량식 고추장 효모의 분포 및 생리특성)

  • Jung, Yoon-Chang;Choi, Won-Jin;Oh, Nam-Soon;Han, Min-Su
    • Korean Journal of Food Science and Technology
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    • v.28 no.2
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    • pp.253-259
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    • 1996
  • To investigate the yeast florae in the traditional and commercial Kochujang, computer identification systems, Vitek, API kit and conventional identification methods were used. Yeast florae of each process were compared and their typical physiological characteristics were also tested. Various process intervals yielded 330 colonies, which resulted in 11 species 184 strains classified. They were identified into Candida glabrata C. guilliermondii. C. humicola. C. rugosa, C. zeylanoides, Cryptococcus uniguttulatus, Pichia farinosa, Rhodotorula glutinis, Saccharomyces cerevisiae and Zygosaccharomyces rouxii. The strains of Candida, Pichia, Saccharomyces and Zygosaccharomyces were existing in both processes. In case of commercial process, the maximum distribution of Z. rouxii and S. cerevisiae were 33% at 15 day fermentation and 13% at 21 day, respectively. The distribution of Candida spp. was gradually decreased throughtout the fermentation period from 40% to 10%. In the traditional process, the maximum distribution of Z. rouxii and S. cerevisiae were 53% after 3 months and 26% after 7 months, respectively, S. cerevisiae and Z. rouxii showed distintive growth pattern at the high concentration of glucose and sodium chloride and played important roles in both processes of fermentation. Physiological tests revealed that only two major yeasts. S. cerevisiae and Z. rouxii, showed vigorous carbon dioxide formation under the tested conditions.

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