Multilocus sequence typing analysis was applied to determine the genotypes of 147 (137 clinical and 10 environmental) Cryptococcus neoformans and three clinical Cryptococcus gattii isolates from 1993 to 2014 in Korea. Among the 137 clinical isolates of C. neoformans, the most prevalent genotype was ST5 (n = 131), followed by ST31 (n = 5) and ST127 (n = 1). Three C. gattii strains were identified as ST57, ST7, and ST113. All environmental isolates were identified as C. neoformans with two genotypes, ST5 (n = 7) and ST31 (n = 3). Our results show that C. neoformans isolates in Korea are genetically homogeneous, and represent a close genetic relationship between clinical and environmental isolates.
Cryptococcus gattii* ; Cryptococcus neoformans* ; Cryptococcus* ; Genotype* ; Korea* ; Multilocus Sequence Typing

China ; epidemiology ; Cryptococcosis ; epidemiology ; Cryptococcus gattii ; Humans

BACKGROUNDInitially, Cryptococcus (C.) neoformans was previously divided into two varieties comprising C. neoformans var. neoformans and C. neoformans var. gattii. Currently, taxonomic studies defined C. neoformans as C. species complex, which contains C. neoformans var. neoformans (serotype D), the hybrid isolates (serotype AD), C. neoformans var. grubii (serotype A) and C. gattii (serotypes B and C). However, Liao and his team once isolated a unique C. gattii isolate, namely strain S8012 with unique phenotype from cerebrospinal fluid (CSF) of a 43-year-old male patient in the Shanghai Changzheng Hospital and described as C. neoformans var. shanghaiensis in 1980s. The aim of this study was to explore the genetic background and polymorphism of Chinese clinical C. gattii isolates.

METHODSS8012 was analyzed as representative strain using the M13-polymerase chain reaction (PCR) fingerprinting pattern and multilocus sequence analysis including internal transcribed spacers of rDNA (ITS region), the intergenic spacer 1 regions (IGS1), RPB1, RPB2, CNLAC1, and TEF1 genes.

RESULTSThe PCR fingerprinting pattern results showed strain S8012 belonged to molecular types VGI, and phylogenetic analysis suggested strain S8012 was grouped into the cluster of C. gattii environmental isolates originated from Eucalyptus camaldulensis trees in Australia.

CONCLUSIONC. gattii isolates from Chinese patients expresses high polymorphism on the phenotype, and molecular type VGI isolates from China have a close genetic relationship with the C. gattii isolates from Australia.

Cryptococcus ; classification ; genetics ; Cryptococcus gattii ; classification ; genetics ; DNA, Fungal ; genetics ; DNA, Ribosomal ; genetics ; Phylogeny ; Polymerase Chain Reaction

Zinc is an essential micronutrient required for many enzymes that play essential roles in a cell. It was estimated that approximately 3% of the total cellular proteins are required for zinc for their functions. Zinc has long been considered as one of the key players in host-pathogen interactions. The host sequesters intracellular zinc by utilizing multiple cellular zinc importers and exporters as a means of nutritional immunity. To overcome extreme zinc limitation within the host environment, pathogenic microbes have successfully evolved a number of mechanisms to secure sufficient concentrations of zinc for their survival and pathogenesis. In this review, we briefly discuss the zinc uptake systems and their regulation in the model fungus Saccharomyces cerevisiae and in major human pathogenic fungi such as Aspergillus fumigatus, Candida albicans, and Cryptococcus gattii.
Aspergillus fumigatus ; Candida albicans ; Cryptococcus gattii ; Fungi* ; Host-Pathogen Interactions ; Humans ; Micronutrients ; Saccharomyces cerevisiae ; Virulence ; Zinc*

BACKGROUNDDespite 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.

METHODSA 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.

RESULTSSix 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.

CONCLUSIONSThere 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.

Animals ; China ; Columbidae ; microbiology ; Cryptococcosis ; microbiology ; Cryptococcus ; isolation & purification ; Cryptococcus gattii ; isolation & purification ; Cryptococcus neoformans ; isolation & purification ; Eucalyptus ; microbiology ; Feces ; microbiology

Cryptococcus gattii causes life-threatening yeast infection in the pulmonary and central nervous systems of humans and animals, and traditionally has been considered to restrict into the tropical and subtropical areas. Despite rare incidence of cryptococcosis caused by C. gattii in Korea, three strains of C. gattii isolated from cryptococcosis patients between 1993 and 2010 were identified. To determine the genetic diversity, 3 strains of C. gattii were typed using PCR fingerprinting with primer M13 and the restriction fragment length polymorphism (RFLP) of orotidine monophosphosphate pyrophosphorylase (URA5) gene. All isolates were identified as serotype B and MATalpha mating type. The molecular types of each strain, on the other hand, turned out to be distinct belonging to VGI, VGII or III types, respectively. Although the travel histories of the patients were not available, clinical C. gattii strains isolated in Korea may represent the diverse molecular types existing worldwide.
Animals ; Central Nervous System ; Cryptococcosis ; Cryptococcus ; Cryptococcus gattii ; Dermatoglyphics ; Genetic Variation ; Hand ; Humans ; Incidence ; Korea ; Molecular Typing ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Sprains and Strains ; Uridine ; Yeasts

OBJECTIVETo understand the species, genotypes and mating types of Cryptococcus neoformans and Cryptococcus gattii isolated from clinical samples in Guigang, Guangxi Zhuang Autonomous Region.

METHODSA total of 20 Cryptococcus strains were isolated from clinical samples in Guigang from 2009 to 2012. The biological identification was conducted by polymerase chain reaction (PCR) to amplify internal transcribed spacer (ITS) sequences. The serotypes and mating types of C. neoformans and C. gattii were identified by PCR with serotype-specific and mating type-specific primers. The genotype was characterized by PCR fingerprinting and URA5 gene restriction fragment length polymorphism (URA5-RFLP). Phenotype study included growth test at 37 °C, melanin production test and urease test.

RESULTSAmong the 20 strains, 19 (95%) were identified as C. neoformans varieties (var.) grubii (serotype A, mating type α, genotype VN I), and only 1 was identified as C. gattii (mating type α, genotype VG I). The results of virulence test showed that all the strains grew well at 37 °C and positive in both urease test and melanin production test.

CONCLUSIONC. neoformans var. grubii (serotype A, genotype VN I and mating type α) was the predominant pathogen causing cryptococcosis in Guigang, and C. gattii strain was also detected.

China ; Cryptococcus gattii ; genetics ; isolation & purification ; pathogenicity ; Cryptococcus neoformans ; genetics ; isolation & purification ; pathogenicity ; Genotype ; Humans ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Virulence

Cryptococcus spp. other than Cryptococcus neoformans or Cryptococcus gattii were previously considered saprophytes and thought to be non-pathogenic to humans. However, opportunistic infections associated with non-neoformans and non-gattii species, such as Cryptococcus laurentii and Cryptococcus albidus, have increased over the past four decades. We experienced a case of cryptococcosis caused by non-neoformans and non-gattii spp. in a 47-year-old female with refractory acute myeloid leukemia after allogeneic hematopoietic stem cell transplantation. The patient underwent salvage chemotherapy with fluconazole prophylaxis and subsequently developed neutropenic fever with multiple erythematous umbilicated papules. A skin biopsy revealed fungal hyphae and repetitive blood cultures showed yeast microorganisms that were identified later as C. laurentii by Vitek-II®. Skin lesions and fever began to improve with conventional amphotericin B therapy. The treatment regimen was continued for 21 days until the disseminated cryptococcosis was completely controlled.
Amphotericin B ; Biopsy ; Cryptococcosis ; Cryptococcus gattii ; Cryptococcus neoformans ; Cryptococcus* ; Drug Therapy ; Female ; Fever ; Fluconazole ; Hematopoietic Stem Cell Transplantation ; Humans ; Hyphae ; Leukemia, Myeloid, Acute* ; Middle Aged ; Opportunistic Infections ; Skin Manifestations* ; Skin* ; Yeasts