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.

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The isolation of Cryptococcosis agents from environmental samples may be difficult due to the presence of groups of fast-growing fungi. We propose a new culture medium based on a modification of Dichloran Rose-Bengal Chloramphenicol Medium (DRBCm) to detect colonies of Cryptococcus neoformans. Our results indicate that DRBCm is superior to the classical Bird Seed Agar in its ability to detect colonies of C. neoformans.

Cryptococcal infection is transmitted by the inhalation of Cryptococcus spp. propagules. Information about the Cryptococcus species inhabiting plants might be clinically relevant due to the epidemiological role of these habitats as possible sources of human infection. The aim of this study was to increase the knowledge about the environmental occurrence of cryptococcosis agents. Hollow tree vegetal debris of nine plant species was sampled quarterly over a 12-month period. Melanized colonies were screened for Cryptococcus neoformans and C. gattii by biochemical tests, followed by URA5-RFLP molecular analysis, M13 fingerprinting assays, and mating-typing with the specific a and α primers. The susceptibility to fluconazole of all of the confirmed species colonies was determined using the AFST-EUCAST broth dilution method. We found that the typical Brazilian flora tree Hymenaea courbaril yielded a high cryptococcal burden (median, 10(2) CFU/g) during the summer, autumn and winter seasons. C. neoformans VNI molecular type MAT alpha was identified in all of the samples. The fingerprinting analyses showed great molecular variability with no correlation with the susceptibility profile to fluconazole (MIC range 4 to ≥64 mg/l). To our knowledge, this study is the first describing the association between C. neoformans and Hymenaea courbaril. These observations extend the known geographic distribution of and substantiate a new urban environmental niche for C. neoformans and also emphasize the genetic diversity of the environmental C. neoformans VNI molecular type isolates.

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