Primeiro registro de Cryptococcus neoformans em excretas de pombos provenientes de locais públicos e residenciais de área metropolitana de Cuiabá, Estado do Mato Grosso, Brasil

RESUMO A criptococose é micose sistêmica potencialmente grave causada por duas espécies do gênero Cryptococcus que acometem tanto homens como animais: Cryptococcus neoformans e C. gattii. São infecções cosmopolitas e emergentes, resultantes da interação do hospedeiro - humano e animal versus meio ambiente. A proposta deste trabalho foi avaliar a ocorrência de C. neoformans em 122 amostras de excretas secas de pombos coletadas em 49 locais na cidade de Cuiabá, Estado do Mato Grosso, Brasil, incluindo: praças públicas (n = 5), igrejas (n = 4), instituições de ensino (n = 3), unidades de saúde (n = 8), áreas abertas exibindo cobertura de amianto (n = 4), conjuntos residenciais domiciliares (n = 23), uma fábrica (n = 1) e um presídio (n = 1). Semeadura de suspensão de amostras em meio ágar niger (NSA), identificação fenotípica por provas bioquímicas e teste em meio de canavanina-glicina-azul de bromotimol, das colônias isoladas com pigmentação marrom escura. Foi também utilizada a técnica da reação em cadeia da polimerase com pares de iniciadores específicos para identificação de C. neoformans. As amostras foram coletadas de julho a dezembro de 2010. Cryptococcus neoformans foi isolado em oito (6,6%) de 122 amostras correspondendo a seis (12,2%) dos 49 sítios analisados. Cryptococcus neoformans associado a excretas de pombos ocorre em áreas de Cuiabá, predominando em residências nas amostras analisadas, constituindo fator de risco potencial para aquisição da doença tanto para indivíduos imunocomprometidos como imunocompetentes.SUMMARY Cryptococcosis is a severe systemic mycosis caused by two species of Cryptococcus that affect humans and animals: C. neoformans and C. gattii. Cosmopolitan and emergent, the mycosis results from the interaction between a susceptible host and the environment. The occurrence of C. neoformans was evaluated in 122 samples of dried pigeon excreta collected in 49 locations in the City of Cuiabá, State of Mato Grosso, Brazil, including public squares (n = 5), churches (n = 4), educational institutions (n = 3), health units (n = 8), open areas covered with asbestos (n = 4), residences (n = 23), factory (n = 1) and a prison (n = 1). Samples collected from July to December of 2010 were seeded on Niger seed agar (NSA). Dark brown colonies were identified by urease test, carbon source assimilation tests and canavanine-glycine-bromothymol blue medium. Polymerase chain reaction primer pairs specific for C. neoformans were also used for identification. Cryptococcus neoformans associated to pigeon excreta was isolated from eight (6.6%) samples corresponding to six (12.2%) locations. Cryptococcus neoformans was isolated from urban areas, predominantly in residences, constituting a risk of acquiring the disease by immunocompromised and immunocompetent individuals

Indoor Dust as a Source of Virulent Strains of the Agents of Cryptococcosis in the Rio Negro Micro-Region of the Brazilian Amazon.

Cryptococcosis, a potentially fatal mycosis in humans, is acquired via exposure to exogenous environmental sources. This study aimed to investigate the frequency, genetic diversity, and virulence of cryptococcal strains isolated from indoor dust in the Rio Negro micro-region of the Brazilian Amazon. A total of 8.9% of the studied houses were positive, recovering nine Cryptococcus neoformans VNI and 16 C. gattii VGII isolates, revealing an endemic pattern in domestic microenvironments. The International Society for Human and Animal Mycology (ISHAM) consensus multilocus sequence typing (MLST) scheme for the C. neoformans/C. gattii species complexes identified two sequence types (STs), ST93 and ST5, amongst C. neoformans isolates and six STs amongst C. gattii isolates, including the Vancouver Island Outbreak ST7 (VGIIa) and ST20 (VGIIb), the Australian ST5, and ST264, ST268 and ST445, being unique to the studied region. Virulence studies in the Galleria mellonella model showed that five C. gattii strains and one C. neoformans strain showed a similar pathogenic potential to the highly virulent Vancouver Island outbreak strain CDR265 (VGIIa). The findings of this study indicate that humans can be exposed to the agents of cryptococcosis via house dust, forming the basis for future studies to analyze the impact of early and continuous exposure to indoor dust on the development of subclinical or clinical infections

Performance of cryptococcal antigen lateral flow assay in serum, cerebrospinal fluid, whole blood, and urine in HIV-infected patients with culture-proven cryptococcal meningitis admitted at a Brazilian referral center

Cryptococcal meningitis is the most common cause of opportunistic meningitis in HIV-infected patients in Brazil and causes unacceptable high mortality rates. In this study, HIV-infected patients with a first episode of culture-proven cryptococcal meningitis in cerebrospinal fluid (CSF) were prospectively included in order to evaluate sensitivity of cryptococcal antigen (CrAg) lateral flow assay (LFA) in serum, CSF, whole blood (fingerstick), and fresh urine. In addition, HIV-infected patients with other neurological confirmed diseases were included in order to evaluate the specificity of CrAg LFA in serum. Twenty patients with cryptococcal meningitis were included and in 19 of them, CrAg LFA in CSF, serum, and whole blood were positive (95% sensitivity). In 18 patients, India ink test was positive in CSF (90% sensitivity), and in 16 cases, CrAg LFA was positive in urine (80% sensitivity). Thirty-six HIV-infected patients with other neurological diseases had negative results of CrAg LFA in serum (100% specificity). In conclusion, CrAg LFA in serum, CSF, and whole blood showed high sensitivity and specificity. Whole blood CrAg LFA seems to be a good and reliable strategy to improve AIDS-related cryptococcal meningitis diagnosis in Brazil

Cryptococcus gattii in North American Pacific Northwest: whole-population genome analysis provides insights into species evolution and dispersal

The emergence of distinct populations of Cryptococcus gattii in the temperate North American Pacific Northwest (PNW) was surprising, as this species was previously thought to be confined to tropical and semitropical regions. Beyond a new habitat niche, the dominant emergent population displayed increased virulence and caused primary pulmonary disease, as opposed to the predominantly neurologic disease seen previously elsewhere. Whole-genome sequencing was performed on 118 C. gattii isolates, including the PNW subtypes and the global diversity of molecular type VGII, to better ascertain the natural source and genomic adaptations leading to the emergence of infection in the PNW. Overall, the VGII population was highly diverse, demonstrating large numbers of mutational and recombinational events; however, the three dominant subtypes from the PNW were of low diversity and were completely clonal. Although strains of VGII were found on at least five continents, all genetic subpopulations were represented or were most closely related to strains from South America. The phylogenetic data are consistent with multiple dispersal events from South America to North America and elsewhere. Numerous gene content differences were identified between the emergent clones and other VGII lineages, including genes potentially related to habitat adaptation, virulence, and pathology. Evidence was also found for possible gene introgression from Cryptococcus neoformans var. grubii that is rarely seen in global C. gattii but that was present in all PNW populations. These findings provide greater understanding of C. gattii evolution in North America and support extensive evolution in, and dispersal from, South America. Importance: Cryptococcus gattii emerged in the temperate North American Pacific Northwest (PNW) in the late 1990s. Beyond a new environmental niche, these emergent populations displayed increased virulence and resulted in a different pattern of clinical disease. In particular, severe pulmonary infections predominated in contrast to presentation with neurologic disease as seen previously elsewhere. We employed population-level whole-genome sequencing and analysis to explore the genetic relationships and gene content of the PNW C. gattii populations. We provide evidence that the PNW strains originated from South America and identified numerous genes potentially related to habitat adaptation, virulence expression, and clinical presentation. Characterization of these genetic features may lead to improved diagnostics and therapies for such fungal infections. The data indicate that there were multiple recent introductions of C. gattii into the PNW. Public health vigilance is warranted for emergence in regions where C. gattii is not thought to be endemic

The status of cryptococcosis in Latin America

Cryptococcosis is a life-threatening fungal infection caused by the encapsulated yeasts Cryptococcus neoformans and C. gattii, acquired from the environment. In Latin America, as occurring worldwide, C. neoformans causes more than 90% of the cases of cryptococcosis, affecting predominantly patients with HIV, while C. gattii generally affects otherwise healthy individuals. In this region, cryptococcal meningitis is the most common presentation, with amphotericin B and fluconazole being the antifungal drugs of choice. Avian droppings are the predominant environmental reservoir of C. neoformans, while C. gattii is associated with several arboreal species. Importantly, C. gattii has a high prevalence in Latin America and has been proposed to be the likely origin of some C. gattii populations in North America. Thus, in the recent years, significant progress has been made with the study of the basic biology and laboratory identification of cryptococcal strains, in understanding their ecology, population genetics, host-pathogen interactions, and the clinical epidemiology of this important mycosis in Latin America.Fil: Firacative, Carolina. University of Sydney; AustraliaFil: Lizarazo, Jairo. Universidad de Pamplona; EspañaFil: Illnait Zaragozí, María Teresa. Tropical Medicine Institute Pedro Kourí; CubaFil: Castañeda, Maria Elizabeth. Instituto Nacional de Salud; Colombia. Latin American Cryptococcal Study Group; BrasilFil: Arechavala, Alicia. Gobierno de la Ciudad de Buenos Aires. Hospital de Infecciosas "Dr. Francisco Javier Muñiz"; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Córdoba, Susana Beatríz. Latin American Cryptococcal Study Group; Brasil. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; ArgentinaFil: Mazza, Mariana. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Taverna, Constanza Giselle. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Isla, Guillermina. Latin American Cryptococcal Study Group; Brasil. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud “Dr. C. G. Malbrán”; ArgentinaFil: Chiapello, Laura Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina. Latin American Cryptococcal Study Group; BrasilFil: Vergara, Mario León Silva. Universidade Federal do Triangulo Mineiro; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Melhem, Marcia S. C.. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Szeszs, Maria Walderez. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Martins, Marilena dos Anjos. Latin American Cryptococcal Study Group; Brasil. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; BrasilFil: Bonfietti, Lucas Xavier. Latin American Cryptococcal Study Group; Brasil. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; BrasilFil: Oliveira, Rogério Antonio de. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Oliveira, Lidiane de. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Santos, Dayane Christine Silva. Latin American Cryptococcal Study Group; Brasil. Governo do Estado de São Paulo. Secretaria da Saúde. Instituto Adolfo Lutz; BrasilFil: Lazera, Marcia S.. Latin American Cryptococcal Study Group; Brasil. Fundación Oswaldo Cruz; BrasilFil: Wanke, Bodo. Fundación Oswaldo Cruz; Brasil. Latin American Cryptococcal Study Group; BrasilFil: Díaz, María Cristina. Latin American Cryptococcal Study Group; Brasil. Universidad de Chile; ChileFil: Escandón, Patricia. Instituto Nacional de Salud; Colombia. Latin American Cryptococcal Study Group; BrasilFil: Noguera, María Clara. Latin American Cryptococcal Study Group; Brasil. Universidad Metropolitana; ColombiaFil: Andreu, Carlos Manuel Fernández. Latin American Cryptococcal Study Group; BrasilFil: Castril­Lón, Laura. Universidad Nacional Autónoma de México; México. Latin American Cryptococcal Study Group; BrasilFil: Bustamante, Beatriz. Hospital Cayetano Heredia. Instituto de Medicina Tropical Alexander von Humboldt; Perú. Hospital Cayetano Heredia; Perú. Latin American Cryptococcal Study Group; BrasilFil: Dolande, Maribel. Universidad Central de Venezuela; Venezuela. Latin American Cryptococcal Study Group; BrasilFil: Ferrara, Giussepe. Universidad Central de Venezuela; Venezuela. Latin American Cryptococcal Study Group; Brasi

Consenso em criptococose - 2008

Univ Sao Paulo, Fac Med, Div Clin Mol Infecciosas, Hosp Clin, Sao Paulo, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Dept Med, Sao Paulo, BrazilUniv Estadual Campinas, Hosp Clin, Inst Infectol Emilio Ribas, Campinas, SP, BrazilFundacao Oswaldo Cruz, Dept Microbiol Immunol & Parasitol, Inst Pesquisa Clin Evandro Chagas, Rio De Janeiro, BrazilUniv Fed Parana, Fac Med, Dept Saude Comunitaria, BR-80060000 Curitiba, Parana, BrazilUniv Fed Rio Grande do Sul, Fac Med, Dept Clin Med, Porto Alegre, RS, BrazilUniv Estadual Campinas, Fac Ciencias Med, Dept Clin Med, Sao Paulo, BrazilInst Doencas Trop Natan Portela, Teresina, PI, BrazilUniv Sao Paulo, Fac Med, Dept Mol Infecciosas & Parasitarias, Sao Paulo, BrazilUniv Estadual Campinas, Fac Ciencias Med, Dept Clin Med, Campinas, SP, BrazilUniv Fed Uberlandia, Fac Med, BR-38400 Uberlandia, MG, BrazilFac Med Triangulo Mineiro, Dept Clin Med, Uberaba, MG, BrazilInst Infectol Emilio Ribas, Sao Paulo, BrazilUniv Estadual Sao Paulo, Fac Med Botucatu, Dept Doencas Trop & Diagnost Imagem, Sao Paulo, BrazilUniv Sao Paulo, Fac Med Ribeirao Preto, Dept Clin Med, Ribeirao Preto, SP, BrazilUniv Fed Sao Paulo, Escola Paulista Med, Dept Med, Sao Paulo, BrazilWeb of Scienc

Ancient dispersal of the human fungal pathogen Cryptococcus gattii from the Amazon rainforest.

Over the past two decades, several fungal outbreaks have occurred, including the high-profile 'Vancouver Island' and 'Pacific Northwest' outbreaks, caused by Cryptococcus gattii, which has affected hundreds of otherwise healthy humans and animals. Over the same time period, C. gattii was the cause of several additional case clusters at localities outside of the tropical and subtropical climate zones where the species normally occurs. In every case, the causative agent belongs to a previously rare genotype of C. gattii called AFLP6/VGII, but the origin of the outbreak clades remains enigmatic. Here we used phylogenetic and recombination analyses, based on AFLP and multiple MLST datasets, and coalescence gene genealogy to demonstrate that these outbreaks have arisen from a highly-recombining C. gattii population in the native rainforest of Northern Brazil. Thus the modern virulent C. gattii AFLP6/VGII outbreak lineages derived from mating events in South America and then dispersed to temperate regions where they cause serious infections in humans and animals

Identification of Novel Hybrids Between Cryptococcus neoformans var. grubii VNI and Cryptococcus gattii VGII

Cryptococcus neoformans and Cryptococcus gattii are pathogenic yeasts causing meningoencephalitis in immunocompromised and immunocompetent hosts. The fungus is typically haploid, and sexual reproduction occurs normally between individuals with opposite mating types, α and a. C. neoformans var. grubii (serotype A) is comprised of molecular types VNI, VNII, and VNB, and C. neoformans var. neoformans (serotype D) contains the molecular type VNIV. Additionally, diploid or aneuploid AD hybrids (VNIII) have been reported. C. gattii contains the molecular types VGI, VGII, VGIII, and VGIV, which encompass both serotypes B and C. To identify possible hybrid strains, URA5-RFLP analysis was performed on 350 globally obtained clinical, environmental, and veterinary isolates. Four clinical isolates from cerebrospinal fluid showed combination patterns of C. neoformans var. grubii and C. gattii: Brazil (n = 2), Colombia (n = 1), and India (n = 1). These strains were monokaryotic and diploid or aneuploid. M13 PCR fingerprinting showed that they contained fragments of both proposed parental groups. Luminex IGS genotyping identified these isolates as hybrids with two different molecular type combinations: three VNI/VGII and one VNI/VGI. Blue color development on CGB agar was delayed in three isolates and absent in one. C. gattii-specific PCR confirmed the presence of C. gattii in the hybrids. CAP59 allele-specific PCR revealed that all the hybrids contained both serotype A and B alleles. Determination of mating-type allelic patterns by PCR revealed that the isolates were αA aB. This is the first study discovering novel natural hybrids between C. neoformans molecular type VNI and C. gattii molecular type VGII