title: Molecular typing of environmental Cryptococcus neoformans/C. gattii species complex isolates from Manaus, Amazonas, Brazil.
abstract: 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.
authors.affiliation: Programa de Pós-Graduação em Saúde Sociedade e Endemias na Amazônia - UFAM and CPqLMD/FIOCRUZ, Manaus, Amazonas, Brazil.
authors.forename: Gleica Soyan Barbosa
authors.initials: GS
authors.lastname: Alves
authors.affiliation: Programa de Pós-Graduação em Medicina Tropical, Universidade do Estado do Amazonas (UEA) and Fundação de Medicina Tropical Dr. Heitor Vieira Dourado (FMT-HVD), Manaus, Amazonas, Brazil.
authors.forename: Ana Karla Lima
authors.initials: AK
authors.lastname: Freire
authors.affiliation: Programa de Pós-Graduação em Medicina Tropical, Universidade do Estado do Amazonas (UEA) and Fundação de Medicina Tropical Dr. Heitor Vieira Dourado (FMT-HVD), Manaus, Amazonas, Brazil.
authors.forename: Amaury Dos Santos
authors.initials: Ados S
authors.lastname: Bentes
authors.affiliation: Centro de Controle de Zoonoses de Manaus, Manaus, Amazonas, Brazil.
authors.forename: José Felipe de Souza
authors.initials: JF
authors.lastname: Pinheiro
authors.affiliation: Instituto Nacional de Pesquisas da Amazônia (INPA), Manaus, Amazonas, Brazil.
authors.forename: João Vicente Braga
authors.initials: JV
authors.lastname: de Souza
authors.affiliation: Laboratório de Micologia do Instituto Nacional de Infectologia Evandro Chagas (INI)/FIOCRUZ, Rio de Janeiro, Brazil.
authors.forename: Bodo
authors.initials: B
authors.lastname: Wanke
authors.affiliation: Laboratório de Microbiologia, Universidade Federal do Amazonas (UFAM), Manaus, Amazonas, Brazil.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Matsuura
authors.affiliation: Centro de Pesquisa Leônidas e Maria Deane (CPqLMD)/FIOCRUZ, Manaus, Amazonas, Brazil.
authors.forename: Ani Beatriz
authors.initials: AB
authors.lastname: Jackisch-Matsuura
mesh_terms: D000666:Amphotericin B / Q000494:pharmacology; D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D001717:Birds / Q000382:microbiology; D001938:Brazil; D003453:Cryptococcosis / Q000382:microbiology; D056285:Cryptococcus gattii / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification* / Q000472:pathogenicity; D003455:Cryptococcus neoformans / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification* / Q000472:pathogenicity; D004197:Disease Reservoirs / Q000382:microbiology; D004783:Environmental Microbiology; D005243:Feces / Q000382:microbiology; D015725:Fluconazole / Q000494:pharmacology; D006801:Humans; D017964:Itraconazole / Q000494:pharmacology; D007654:Ketoconazole / Q000494:pharmacology; D058889:Molecular Typing / Q000379:methods*; D016533:Mycological Typing Techniques / Q000379:methods; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D014197:Trees / Q000382:microbiology
chemical_list: D000935:Antifungal Agents; D017964:Itraconazole; D000666:Amphotericin B; D015725:Fluconazole; D007654:Ketoconazole
keywords: Amazonas; Brazil; Cryptococcus gattii; Cryptococcus neoformans; Manaus; environmental isolation; genotyping