Cryptococcus neoformans-Cryptococcus gattii species complex: an international study of wild-type susceptibility endpoint distributions and epidemiological cutoff values for fluconazole, itraconazole, posaconazole, and voriconazole.

Epidemiological cutoff values (ECVs) for the Cryptococcus neoformans-Cryptococcus gattii species complex versus fluconazole, itraconazole, posaconazole, and voriconazole are not available. We established ECVs for these species and agents based on wild-type (WT) MIC distributions. A total of 2,985 to 5,733 CLSI MICs for C. neoformans (including isolates of molecular type VNI [MICs for 759 to 1,137 isolates] and VNII, VNIII, and VNIV [MICs for 24 to 57 isolates]) and 705 to 975 MICs for C. gattii (including 42 to 260 for VGI, VGII, VGIII, and VGIV isolates) were gathered in 15 to 24 laboratories (Europe, United States, Argentina, Australia, Brazil, Canada, Cuba, India, Mexico, and South Africa) and were aggregated for analysis. Additionally, 220 to 359 MICs measured using CLSI yeast nitrogen base (YNB) medium instead of CLSI RPMI medium for C. neoformans were evaluated. CLSI RPMI medium ECVs for distributions originating from at least three laboratories, which included ≥95% of the modeled WT population, were as follows: fluconazole, 8 µg/ml (VNI, C. gattii nontyped, VGI, VGIIa, and VGIII), 16 µg/ml (C. neoformans nontyped, VNIII, and VGIV), and 32 µg/ml (VGII); itraconazole, 0.25 µg/ml (VNI), 0.5 µg/ml (C. neoformans and C. gattii nontyped and VGI to VGIII), and 1 µg/ml (VGIV); posaconazole, 0.25 µg/ml (C. neoformans nontyped and VNI) and 0.5 µg/ml (C. gattii nontyped and VGI); and voriconazole, 0.12 µg/ml (VNIV), 0.25 µg/ml (C. neoformans and C. gattii nontyped, VNI, VNIII, VGII, and VGIIa,), and 0.5 µg/ml (VGI). The number of laboratories contributing data for other molecular types was too low to ascertain that the differences were due to factors other than assay variation. In the absence of clinical breakpoints, our ECVs may aid in the detection of isolates with acquired resistance mechanisms and should be listed in the revised CLSI M27-A3 and CLSI M27-S3 documents.

Environmental isolation and characterisation of Cryptococcus species from living trees in Havana city, Cuba.

Cryptococcus isolates from Cuban patients were identified as C. neoformans var. grubii. Although this species has since long been associated with bird droppings, a recent genotyping study provided strong evidence for additional origins of exposure. We sampled different species of trees in Havana, Cuba to identify other potential sources of exposure to this fungus. A total of 662 samples were collected from 331 trees and cacti from Havana, Cuba. Initial selection of the isolates was carried out by conventional techniques. Isolates were further characterised using a combination of AFLP analysis and DNA sequence analysis. Identification by conventional methods yielded 121 C. neoformans and 61 C. gattii isolates. Molecular analyses showed that none of these isolates was C. gattii and only one isolate proved to be C. neoformans var. grubii. A total of 27 different other species were identified. The most prevalent species was C. heveanensis (33%). Sixty-five unidentifiable isolates segregated into ten potentially novel species. Conventional cultivation methods have a low specificity for C. neoformans complex and molecular analyses need to be applied to confirm identification of isolates from environmental sources. Environmental niches responsible for most of human cryptococcal infections in Cuba remain to be identified.

[Isolation of anti-Cryptococcus neoformans immunoglobulin G]. / Obtención de inmunoglobulina G anti-Cryptococcus neoformans.

Two immunization schemes were applied to rabbits with an autoctonous strain aimed at obtaining an anti-Crytococcus neoformans IgG. Whole cells treated with formalin were used. Dosages and immunization routes were different. The sera obtained were titered and that with the highest titer was selected (1:1 024 by the laminar agglutination method) and purified by saline precipitation and ion exchange chromatography. An IgG of high purity and biological activity was obtained.

[Minimum inhibitory concentration of amphotericin B in yeasts of medical interest]. / Determinación de la concentración mínima inhibitoria de anfotericina B en levaduras de interés médico.

In order to know the sensitivity of Candida and Crytococcus to amphotericin B, main drug for the treatment od systemic mycosis, it was determined the minimum inhibitory concentration (MIC) in 90 clinical isolates of Candida and Crytococcus by a micromethod for the broth dilution. According to the results, Crytococcus neoformas was more sensitive then Candida albicans (geometrical means 0.24 and 0.41 respectively). Only one resistant strain was found (CMI = 16 micrograms/mL), corresponding to the Candida krusei species. The introduction of this technique in the Mycology Laboratory of the "Pedro Kourí" Institute of Tropical Medicine will allow to establish the sensitivity patterns and to detect the possible appearance of resistance in the main species of pathogenic fungus for men in our environment.

[The identification of Cryptococcus neoformans var. neoformans in Cuban clinical isolates]. / Identificación de Cryptococcus neoformans var. neoformans en aislamientos clínicos cubanos.

50 strains of Cryptococcus neoformans of clinical origin were studied by using the canavanine-glycine-bromothymol blue (CGB) medium. 56% of the strains were isolated from AIDS patients, and 16% belonged to patients with kidney transplantation. 90% of the samples were obtained from the samples of cerebrospinal fluid, which corresponded to the classical form of presentation of cryptococcosis. All the strains were identified as C. neoformans var. neoformans, coinciding with previous reports made in Cuba. Knowing the varieties of C. neoformans, inferences can be drawn on the epidemiology, clinics and response to the treatment of cryptococcosis.