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

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

Aged ; Cryptococcosis ; microbiology ; pathology ; surgery ; Cryptococcus neoformans ; isolation & purification ; Follow-Up Studies ; Humans ; Laryngeal Diseases ; microbiology ; pathology ; surgery ; Male

Pulmonary infection by capsule-deficient Cryptococcus neoformans (CDCN) is a very rare form of pneumonia and it is seldom seen in the immunocompetent host. The authors experienced a case of pulmonary cryptococcosis by CDCN in 25-year-old woman who was without any significant underlying disease. The diagnosis was made from the percutaneous lung biopsy and special tissue staining, including Fontana-Masson silver (FMS) staining. Fungal culture confirmed the diagnosis afterward. Her clinical and radiologic features improved under treatment with fluconazol. It's known that CDCN is not so readily confirmed because fungal culture does not always result in growth of the organism and the empirical fungal stain is not helpful for the differentiation between CDCN and the other infections that are caused by the nonencapsulated yeast-like organisms. In this report, we emphasize the diagnostic value of performing FMS staining for differentiating a CDCN infection from the other confusing nonencapsulated yeast-like organisms.
Silver Nitrate ; Lung Diseases/*diagnosis/*microbiology ; Humans ; Female ; Cryptococcus neoformans/*isolation & purification ; Cryptococcosis/*diagnosis/microbiology ; Cough ; Chest Pain ; Adult

BACKGROUND: We utilized results from the ARTEMIS DISK Global Antifungal Surveillance Program to evaluate the species distribution and fluconazole and voriconazole susceptibilities of yeast isolates from clinical specimens in South Korea from 2001 to 2007. METHODS: Data were collected on 5,665 yeast isolates from all body sites at three locations. All investigators tested clinical yeast isolates using the CLSI M44-A disk diffusion method. Test plates were automatically read and results were recorded using the BIOMIC image analysis plate reader system (Giles Scientific, USA). Species, drug, zone diameter, susceptibility category, and quality control results were collected quarterly via e-mail for analysis. RESULTS: Candida albicans was the most common isolate, but a progressive increase in non-C. albicans Candida and noncandidal yeast species has been observed in recent years. The overall percentages of isolates in each category (susceptible, susceptible dose dependent, and resistant) were 98.8%, 0.5%, and 0.7% and 99.2%, 0.2%, and 0.6% for fluconazole and voriconazole, respectively. Candida of 3 species exhibited decreased susceptibility to fluconazole (<90% S) in the order of that seen with the resistant (R) species: C. krusei, C. guilliermondii, and C. glabrata. Emerging resistance to fluconazole or voriconazole was documented among isolates of Cryptococcus neoformans, Trichosporon spp., and Rhodotorula spp. CONCLUSIONS: The species distribution and antifungal susceptibilities of yeasts may differ according to specimen type, testing method, hospital, and geographic region. Therefore, further large-scaled, long-term surveillance studies are needed to isolate yeasts and to confirm the species distribution and antifungal susceptibilities of yeast isolates from clinical specimens in Korea.
Antifungal Agents/*pharmacology ; Candida/isolation & purification ; Cryptococcus neoformans/isolation & purification ; Disk Diffusion Antimicrobial Tests ; *Drug Resistance, Fungal ; Fluconazole/pharmacology ; Hospitals ; Humans ; Pyrimidines/pharmacology ; Republic of Korea ; Rhodotorula/isolation & purification ; Triazoles/pharmacology ; Trichosporon/isolation & purification ; Yeasts/*drug effects/isolation & purification

OBJECTIVETo investigate the chemical constituents of Pyrola calliatha.

METHODThe chemical constituents were isolated by various column chromatographic methods. The structures were identified by spectral data.

RESULTTen compounds were isolated and identified as chimaphilin (1), uvaol(2), ursolic acid (3), 2beta,3beta,23-trihydroxy-12-ene-28-ursolic acid (4), daucosterol (5), 2alpha,3beta,23,24-tetrahydroxy-12-ene-28-ursolic acid (6), emodin (7), gallic acid (8), monotropein (9), adenosine (10).

CONCLUSIONCompounds 2,4,6,7,10 were obtained from this genus for the first time, compounds 5, 9 were obtained from this species for the first time. Antifungal activity of compounds 1-4, 6-9 were evaluated. Compound 1 showed the strong activity.

Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Cryptococcus neoformans ; drug effects ; Iridoids ; chemistry ; isolation & purification ; pharmacology ; Naphthoquinones ; chemistry ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Pyrola ; chemistry ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

OBJECTIVETo investigate diagnostic histopathology and ultrastructure features of primary pulmonary cryptococcosis (PC).

METHODSClinical data and pathologic findings of 27 cases of PC were retrospectively reviewed, light and electron microscopic evaluations and histochemistry stain studies were performed.

RESULTSThe specimens consisted of 2 fine-needle aspiration lung biopsies and 25 cases of open lung biopsies. Cryptococcosis granuloma formation was identifiable by histopathological examination in 25 of 27 cases, with gum-like lesion and fungi in the remaining 2 cases. The detection rates of cryptococcus neoformans (CN) by mucicarmine (MC), periodic acid-Schiff (PAS), alcian blue (AB) and Grocott methenamine-silver (GMS) were 87.0% (20/23), 100% (27/27), 66.7% (18/27), and 100% (23/23) respectively. Under the electron microscope, most CN had a simple structure with a few organelles. The capsule was seen in all organisms. A percentage of the organisms showed nuclei, nucleoli, mitochondria and vacuoles. The detection rate of CN by EM was 91.7% (11/12).

CONCLUSIONSThe clinical manifestation and imaging of PC are nonspecific for PC. Lung biopsy is the major diagnostic modality. The detection rate by electron microscopy was quite high. Therefore, a correct diagnosis of pulmonary cyrptococcosis should rely on the combination of histopathological evaluation, histochemistry staining and/or electron microscopic examination.

Adult ; Aged ; Biopsy, Fine-Needle ; Cryptococcosis ; microbiology ; pathology ; Cryptococcus neoformans ; isolation & purification ; ultrastructure ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Lung ; pathology ; ultrastructure ; Lung Diseases, Fungal ; classification ; microbiology ; pathology ; Male ; Microscopy, Electron ; Middle Aged ; Retrospective Studies

OBJECTIVETo research the isolation method, identification and screen for bioactivities endophytic fungi from ginkgo.

METHODEndophytic fungi from ginkgo were separated. By means of microdilution method, activities of endophytes against pathogenic fungi were tested. Then, using DPPH, the antioxidant activities were measured.

RESULTNine strains (16.1%) showed antifungal activities against Candida albicans, Cryptococcus neoformans, Trichophyton rubrum and Aspergillus fumigatus. Among these bioactive strains, the growth of T. rubrum was strongly inhibited by T-1-2-1, as the MIC80 was equal to fluconazole, the positive control. Five strains (8.9%) showed antioxidant activities. Among them sample T-3-2-2 and T-6-5-7 showed the strongest antioxidant activities.

CONCLUSIONEndophytic fungi of ginkgo would be potential and rich resources for drug development.

Antifungal Agents ; pharmacology ; Aspergillus fumigatus ; drug effects ; Candida albicans ; drug effects ; Cryptococcus neoformans ; drug effects ; Fluconazole ; pharmacology ; Fungi ; chemistry ; drug effects ; isolation & purification ; Ginkgo biloba ; microbiology ; Microbial Sensitivity Tests ; Trichophyton ; drug effects

OBJECTIVETo explore the pharmacokinetics of amphotericin B (AMB) in the cerebrospinal fluid (CSF) during continuous intrathecal administration of AMB for treatment of cryptococcal neoformans meningitis (CNM).

METHODSThe concentration of AMB in the CSF was measured using reversed phase high performance liquid chromatography (RP-HPLC) in 3 patients receiving continuous intrathecal infusion of AMB for CNM.

RESULTSAMB concentrations in the CSF of the 3 patients exceeded the minimal inhibitory concentration (MIC) of AMB against Cryptococcus neoformans. The concentration-time curve showed that AMB concentration in the CSF underwent obvious variations on the first day of intrathecal infusion and after additional AMB doses, but maintained a stable level (0.61-1.21 µg/ml) on the next day.

CONCLUSION[corrected] Continuous intrathecal administration of AMB can enhance the drug concentration in the CSF and maintain a stable and effective drug level for treatment of CNM.

Adolescent ; Amphotericin B ; administration & dosage ; pharmacokinetics ; Antifungal Agents ; administration & dosage ; pharmacokinetics ; Cerebrospinal Fluid ; metabolism ; Cryptococcus neoformans ; isolation & purification ; Female ; Humans ; Infusions, Spinal ; methods ; Male ; Meningitis, Cryptococcal ; drug therapy ; metabolism

A 36-year-old homosexual Mexican man was admitted to our hospital, with a 30-day history of fever and headache. Upon cerebrospinal fluid examination, the patient's white blood cell count was 1,580/L, total protein was 26 mg/dL, sugar was 17 mg/dL, and his intracranial pressure was 23 cmH2O. The patient was diagnosed with HIV (Human Immunodeficiency Virus) infection by serum Western blotting. Cryptococcus neoformans was isolated in cultures of the patient's blood and cerebrospinal fluids. Chest computerized tomography revealed diffuse reticulonodular infiltration and a ground-glass appearance in both perihilar regions, suggestive of either Pneumocystis carinii pneumonia or cryptococcal pneumonia. On the patient's 6th day in our hospital, bronchoalveolar lavage and transbronchial lung biopsy were conducted via bronchoscopy, and a pathologic examination of lung biopsy specimens revealed signs of cryptococcal pneumonia. This patient died on his 14th day in our hospital, as the result of acute respiratory failure, associated with cryptococcal pneumonia and disseminated cryptococcosis.
Respiratory Insufficiency/*diagnosis/etiology/*microbiology ; Pneumonia/*diagnosis/*microbiology ; Male ; Humans ; Fatal Outcome ; Cryptococcus neoformans/*isolation & purification ; Cryptococcosis/*diagnosis/microbiology ; Bronchoalveolar Lavage ; Blotting, Western ; Adult ; Acute Disease ; AIDS-Related Opportunistic Infections/complications/*microbiology