Molecular typing of environmental Cryptococcus neoformans/C. gattii species complex isolates from Manaus, Amazonas, Brazil

• Published in: Mycoses Vol. 59; no. 8; pp. 509 - 515
• Main Authors: Alves, Gleica Soyan Barbosa, Freire, Ana Karla Lima, Bentes, Amaury dos Santos, Pinheiro, José Felipe de Souza, de Souza, João Vicente Braga, Wanke, Bodo, Matsuura, Takeshi, Jackisch-Matsuura, Ani Beatriz
• Format: Journal Article
• Language: English
• Published: Germany Blackwell Publishing Ltd 01-08-2016
• Subjects: Amazonas; Amphotericin B - pharmacology; Animals; Antifungal Agents - pharmacology; Birds - microbiology; Brazil; Cryptococcosis - microbiology; Cryptococcus gattii; Cryptococcus gattii - classification; Cryptococcus gattii - genetics; Cryptococcus gattii - isolation & purification; Cryptococcus gattii - pathogenicity; Cryptococcus neoformans; Cryptococcus neoformans - classification; Cryptococcus neoformans - genetics; Cryptococcus neoformans - isolation & purification; Cryptococcus neoformans - pathogenicity; Disease Reservoirs - microbiology; environmental isolation; Environmental Microbiology; Feces - microbiology; Fluconazole - pharmacology; genotyping; Humans; Itraconazole - pharmacology; Ketoconazole - pharmacology; Manaus; Molecular Typing - methods; Mycological Typing Techniques - methods; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Trees - microbiology; Cryptococcus gattii; environmental isolation; Amazonas; Cryptococcus neoformans; genotyping; Brazil; Manaus
• ISSN: 0933-7407; 1439-0507
• DOI: 10.1111/myc.12499

Summary
Cryptococcus neoformans and Cryptococcus gattii are the main causative agents of cryptococcosis, a systemic fungal disease that affects internal organs and skin, and which is acquired by inhalation of spores or encapsulated yeasts. It is currently known that the C. neoformans/C. gattii species complex has a worldwide distribution, however, some molecular types seem to prevail in certain regions. Few environmental studies of Cryptococcus have been conducted in the Brazilian Amazon. This is the first ecological study of the pathogenic fungi C. neoformans/C. gattii species complex in the urban area of Manaus, Amazonas, Brazil. A total of 506 samples from pigeon droppings (n = 191), captive bird droppings (n = 60) and tree hollows (n = 255) were collected from June 2012 to January 2014 at schools and public buildings, squares, pet shops, households, the zoo and the bus station. Samples were plated on niger seed agar (NSA) medium supplemented with chloramphenicol and incubated at 25°C for 5 days. Dark‐brown colonies were isolated and tested for thermotolerance at 37°C, cycloheximide resistance and growth on canavanine‐glycine‐bromothymol blue agar. Molecular typing was done by PCR‐RFLP. Susceptibility to the antifungal drugs amphotericin B, fluconazole, itraconazole and ketoconazole was tested using Etest® strips. In total, 13 positive samples were obtained: one tree hollow (C. gattiiVGII), nine pigeon droppings (C. neoformansVNI) and three captive bird droppings (C. neoformansVNI). The environmental cryptococcal isolates found in this study were of the same molecular types as those responsible for infections in Manaus.