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

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

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

Genotype and mating type distribution within clinical Cryptococcus neoformans and Cryptococcus gattii isolates from patients with cryptococcal meningitis in Uberaba, Minas Gerais, Brazil

We molecularly characterized 81 cryptococcal isolates recovered from cerebrospinal fluid samples of 77 patients diagnosed between 1998 and 2007 as having cryptococcal meningitis in Uberaba Minas Gerais, Brazil. Fifty-seven (74%) were male with a mean age 35.6 years. Seventy-two (88.9%) of the isolates were from 68 AIDS patients and cryptococcosis was the first AIDS-defining condition in 38 (55.9%) patients. Cryptococcosis and AIDS were simultaneously diagnosed in 25 (65.8%) of these 38 patients. Genotypes were characterized through the use of URA5 restriction fragment length polymorphisms analysis, the genetic variability was determined using PCR-fingerprinting with the minisatellite-specific primer M13, and the mating type and serotypes were established by PCR. Seventy-six of the 81 isolates were Cryptococcus neoformans (93.8%), while the remaining five were C. gattii (6.1%), but all were mating type a. C. neoformans isolates were genotype VNI (serotype A), while C. gattii isolates were VGII. Four of the latter isolates were identical, but only two were from AIDS patients. Six of the nine isolates from non-AIDS patients were VNI. PCR fingerprints of the isolates from two of the three AIDS patients with clinical relapse were 100% identical. The predominance of VNI and mating type a is in accordance with data from other parts of the world. The occurrence of VGII in Minas Gerais indicates a geographical expansion within Brazil.Fundacao de Amparo a Pesquisa de Minas Gerais (FAPEMIG)[CDS-1881/6

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

Manifestações neurológicas nas endocardites infecciosas - revisão de 103 casos estudados - 84 casos do Hospital dos Servidores do Estado-R.J. (1947-1978) e 19 casos do Hospital Estadual São Sebastião-R.J. (1975-1978)

Os autores fazem uma revisão de 103 casos de endocardites infecciosas, dos quais 52 apresentaram manifestações neurológicas. Dos 52 que não tiveram problemas neurológicos, 39 faleceram 175%), enquanto que dos 51 que não tiveram alterações do sistema nervoso, apenas 18 evoluíram para o óbito 135,3%). A endocardite infecciosa com manifestações neurológicas, na presente série, foi dominante no sexo masculino. Os grupos etários mais envolvidos foram as crianças, adolescentes e adultos jovens. Houve outro pico importante em pacientes acima de 50 anos. Os critérios para o diagnóstico de endocardite infecciosa utilizados no presente estudo, foram os clássicos, acrescidos de outros considerados de extrema utilidade, à medida que a experiência foi sendo acumulada, acompanhando paralelamente as modificações dos padrões etiológicos, assim como as alterações dos espectros clínicos da doença, conseqüentes a muitos fatores aqui discutidos. Os principais distúrbios neurológicos observados foram as manifestações meningeias, 25/52 148,1%), alterações do comportamento, 20/52 (38,4%), fenômenos motores (paralisias e paresias), 20/52 (38,4%). Alterações do nível de consciência (torpor ou coma), ao lado de outras menos comuns, como cefaléia 16/52 (26,9%), convulsões, 10/52 (19,2%), afasia 7/52 (13,5%) e manchas de Roth, 7/53 (13,5%). As manifestações neurológicas frequentemente foram múltiplas em um mesmo paciente. Nas formas de endocardite, foram em muitos casos praticamente as alterações que abriram e dominaram a cena clínica. As mais comuns foram: síndrome meningeia, síndrome vascular e encefalopatia tóxica. Foi difícil, em conseqüência de múltiplas manifestações neurológicas num mesmo paciente, estabelecer critérios entre a lesão neurológica e o prognóstico, embora o coma profundo, as convulsões, os distúrbios motores acentuados, a meningite e as alterações do comportamento, isoladamente ou em associação, façam com que o mesmo seja bem mais sério. O germe mais encontrado em nossa série foi oStaphylococcus aureus, relacionado com formas agudas da infecção endocárdica, aliado a processos destrutivos valvulares e a sérias alterações neurológicas tais como meningite, encefalite, infartos, hemorragias e abcessos cerebrais. Os principais achados neurológicos referentes à patologia, no estudo de 15 casos, são apresentados ao lado de conclusões tiradas de reflexões sobre o material analisado e da experiência vivida. Uma revisão da literatura é feita desde os trabalhos iniciais sobre a doença até os dias de hoje, ficando evidenciada a importância do tema pelas grandes contribuições apresentadas pelos diferentes autores.<br>The authors make a revision of 103 cases of infectious endocarditis, 52 of which brought about neurologic manifestations. In the 52 cases showing neurologic problems, 39 died (75%), while in the other 51 cases showing no alterations of the nervous system only 18 evolved to death (35,3%). The present series of infectious endocarditis with neurologic manifestations was dominat in the mate sex. The more affected age groups were children, adolescents and young adults. Another significant peak was shown by patients more than 50 years old. For the diagnosis of infectious endocarditis in the present study, classical criteria were adopted in addition to a few other ones which were recognized to be extremely useful in the process of accumulating experience, as paralel observation was applied to those changes of etiological patterns and to those alterations of clinical specters of the disease, which resulted from many of the factors here described. The major neurologic diserders detected were meningeal manifestations, 25/52 (48,1%), behavior alterations, 20/52 (38,4%) motor phenomena (paralysis and paresis), 20/52 (38,4%), alterations of the level of consciousness (torpor or coma), plus a few less commonones, such as cephalea, 16/52 (30,7%), alterations of cranial pai rs, 14/52 (26,9%), convulsions, 10/52 (19,2%),aphasia, 7/52 (13,5%). Multiple neurologic manifestations were frequent in the same patient. These alterations were practically the opening and dominant feature of the clinical scene in many cases of acute endocardites. Most common were: meningeal syndrome, vascular syndrome and toxic encephalopathy. Due to multiple neurologic manifestations in the same patient it was difficult to establish criteria linking the neurologic lesion to the prognostic although in states of profound coma, convulsion, pronounced motor disorders, meningitis and behavior alterations, isolated or in association, this prognostic becomes rather more serious. The germ more often found in our series was the staphylococcus aureus, related to acute forms of endocardiac infection, associated with valve destructive process and serious neurologic alterations, such as meningitis, encephalitis, infarcts, hemorrhages and cerebral abscesses. A description is given of the major neurologic findings related to the pathology and of the conclusions draw upon reflexion over the material analysed and the experience collected. A review of the existing literature on the disease, from the initial to the lates studies, clearly points out to the importance of the theme to which different authors have made remarkable contributions