Investigação de surto de Histoplasmose pulmonar aguda entre bombeiros em Brazlândia, Distrito Federal, 2017 / Investigation of acute histoplasmosis outbreak among firefighters, Distrito Federal, 2017

Objetivos: Investigar o surto de histoplasmose em bombeiros no Distrito federal/DF, ocorrido em junho de 2017. Métodos: Realizou-se um estudo de coorte por meio das entrevistas realizadas com os bombeiros mediante um questionário semiestruturado. Considerou-se infectado o bombeiro que apresentou tomografia de tórax sugestiva de histoplasmose ou reagente nos testes de imunodifusão e/ou Western blot. Coletou-se amostra ambiental e realizou-se Nested PCR específico para Histoplasma capsulatum. Resultados: Entre 35 bombeiros, 94,3% eram homens; com a mediana de idade de 37 (24-45) anos, 28 foram classificados como infectados. A média de permanência dentro da caverna foi 25 minutos. O fator de risco associado à infecção foi o ato de entrar na caverna (RR=3,86; RA=71,6; p<0,02). Entre 14 amostras ambientais, 50% foram positivas para H. capsulatum. Conclusão: Confirmou-se o surto de histoplasmose de bombeiros em Brazlândia-DF, e foram tomadas ações como a interdição da caverna e o tratamento dos bombeiros.

Objectives: To nvestigate the outbreak of histoplasmosis in firefighters in Federal District/DF, which occurred in June 2017. Methods: A cohort study was conducted through interviews with firefighters by means of a semi-structured questionnaire. Firefighters who presented chest tomography suggestive of histoplasmosis or reacted to immunodiffusion and/or Western blot tests were considered infected. Environmental samples were collected and Nested PCR specific for Histoplasma capsulatum was performed. Results: Among 35 firefighters, 94.3% were men; with a median age of 37 (24-45) years, 28 were classified as infected. The average length of stay inside the cave was 25 minutes. The risk factor associated with infection was the act of entering the cave (RR = 3.86, RA = 71.6, p <0.02). Among 14 environmental samples, 50% were positive for H. capsulatum. Conclusion: The outbreak of histoplasmosis in firefighters in Brazlândia-DF was confirmed, and actions were taken such as banning the cave and treating the firefighters

Paracoccidioides brasiliensis habitat: far beyond armadillo burrows?

Paracoccidioides spp. isolation from environmental samples is rare and hardly reproducible. Molecular techniques have facilitated the fungal detection. However, it can be still difficult. Some strategies to enhance the capacity of DNA detection have been adopted, including the analysis of soil samples belonging to the habitat of animals from which Paracoccidioides spp. have already been isolated, notably armadillo burrows. To date, the detection of Paracoccidioides spp. has not yet been reported from outbreak hotspots. Clusters and outbreaks of acute paracoccidioidomycosis (PCM), usually a more severe clinical form, have currently occurred in urban areas being associated to climate changes, deforestation, and great constructions. These occurrences potentially signalise the fungus' environmental niche, a riddle not yet solved. The authors performed an environmental investigation in a deeply disturbed area, after a highway construction in Rio de Janeiro, Brazil, where a recent outbreak of acute PCM occurred. Specific DNA sequences of Paracoccidioides brasiliensis were detected in shallow soil samples around the highway, reinforcing the association between the road construction and this PCM outbreak.

FIRST REPORT ON Cryptococcus neoformans IN PIGEON EXCRETA FROM PUBLIC AND RESIDENTIAL LOCATIONS IN THE METROPOLITAN AREA OF CUIABÁ, STATE OF MATO GROSSO, 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

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. .

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. .

Cryptococcus gattii VGI and Cryptococcus neoformans VNI Associated with Wood Decay in Ficus Hollow Trees in Rio de Janeiro, Brazil.

Aim: Considering the geographic expansion of Cryptococcus gattii, the aim of this study was to investigate hollows of living trees as a reservoir of C. gattii in Rio de Janeiro, Brazil. Place and Duration of the Study: In an urban quarter of Rio de Janeiro city, 80 samples of decaying wood were collected. In addition, 85 decaying wood samples were collected in the wild rainforest. The samples were analyzed at the Mycology Laboratory, Evandro Chagas Clinical Research Institute, Oswaldo Cruz Foundation, from 2008-2010. Methodology: Samples were collected by scraping the inner decaying wood of the hollows of the trunks of each tree. Pathogenic Cryptococcus species were identified by: brown colonies on niger seed agar (NSA) medium, thermotolerance at 35ºC, cycloheximide sensitivity, carbon and nitrogen assimilation tests performed by 32-Vitek System (Vitek ICB, bioMeriux, Durham, EUA). Canavanine-glycine-bromothymol blue medium (CGB) was used to determine the species of the isolates and the genotypes were determined by restriction fragment length polymorphism of URA5 gene. Results: After plating the samples on NSA, 584 colonies were obtained from the urban quarter. C. gattii VGI was identified in 98% of colonies, followed by C. neoformans VNI 2%. The positivity of the urban area was 7.8%. The concentrations of the fungi in hollows of ficus trees ranged from 50 to 56,250 colony-forming units per gram of sample (CFU/g). Conclusions: For the first time in Rio de Janeiro C. gattii VGI was isolated in a hollow of living tree.

Cryptococcus neoformans carried by Odontomachus bauri ants

Cryptococcus neoformans is the most common causative agent of cryptococcosis worldwide. Although this fungus has been isolated from a variety of organic substrates, several studies suggest that hollow trees constitute an important natural niche for C. neoformans. A previously surveyed hollow of a living pink shower tree (Cassia grandis) positive for C. neoformans in the city of Rio de Janeiro, Brazil, was chosen for further investigation. Odontomachus bauri ants (trap-jaw ants) found inside the hollow were collected for evaluation as possible carriers of Cryptococcus spp. Two out of 10 ants were found to carry phenoloxidase-positive colonies identified as C. neoformans molecular types VNI and VNII. The ants may have acted as a mechanical vector of C. neoformans and possibly contributed to the dispersal of the fungi from one substrate to another. To the best of our knowledge, this is the first report on the association of C. neoformans with ants of the genus Odontomachus.

Genotypes of Cryptococcus neoformans and Cryptococcus gattii as agents of endemic cryptococcosis in Teresina, Piauí (northeastern Brazil)

Throughout Brazil, Cryptococcus neoformans is the cause of cryptococcosis, whereas Cryptococcus gattii is endemic to the northern and northeastern states. In this study, the molecular types of 63 cryptococcal isolates recovered from the cerebrospinal fluid of meningitis patients diagnosed between 2008-2010 in Teresina, Piauí, Brazil, were analysed. Out of the 63 patients, 37 (58.7 percent) were human immunodeficiency virus (HIV)-positive and 26 (41.3 percent) were HIV-negative. URA5-restriction fragment length polymorphism analysis identified 37/63 (58.7 percent) isolates as the C. neoformans VNI genotype, predominantly in HIV-positive patients (32/37, 86.5 percent), and 24/63 (38.1 percent) as the C. gattii VGII genotype, mostly in HIV-negative patients (21/26, 80.8 percent). The occurrence of C. gattii VGII in six apparently healthy children and in seven adolescents/young adults in this region reaffirms the endemic occurrence of C. gattii VGII-induced primary cryptococcosis and early cryptococcal infection. Lethality occurred in 18/37 (48.6 percent) of the HIV-positive subjects and in 13/26 (50 percent) of the HIV-negative patients. Our results provide new information on the molecular epidemiology of C. neoformans and C. gattii in Brazilian endemic areas.

Cryptococcus gattii molecular type VGII as agent of meningitis in a healthy child in Rio de Janeiro, Brazil: report of an autochthonous case / Cryptococcus gattii tipo molecular VGII como agente causador de meningoencefalite em criança saudável no Rio de Janeiro, Brasil: relato de um caso autóctone

Cryptococcus gattii causes meningoencephalitis in immunocompetent hosts, occurring endemically in some tropical and subtropical regions. Recently, this fungus was involved in an outbreak in Vancouver Island and British Columbia (Canada). In this temperate region, the VGII type is predominant. The paper describes an autochthonous case of meningoencephalitis by C. gattii VGII in a previously health child in Rio de Janeiro, considered nonendemic region of Brazil. The fungus was identified by biochemical tests and the molecular type was determined by URA5-RFLP. The present report highlights the need for clinical vigilance for primary cryptococcal meningitis in nonendemic areas.

Cryptococcus gattii é causa de meningoencefalite em hospedeiros imunocompetentes, ocorrendo endemicamente em regiões tropicais e subtropicais. Recentemente foi causador de surtos na Ilha de Vancouver e na Columbia Britânica (Canadá). Nesta região de clima temperado, o tipo VGII é predominante. Relatamos um caso de meningoencefalite pelo C.gattii tipo VGII acometendo criança previamente saudável autóctone do Rio de Janeiro, região não endêmica do Brasil. O agente foi identificado por testes bioquímicos e o tipo molecular determinado através de URA5-RFLP. O presente relato enfatiza a necessidade de vigilância clínica para a meningite criptocóccica primária em áreas não endêmicas.

First isolation of Cryptococcus gattii molecular type VGII and Cryptococcus neoformans molecular type VNI from environmental sources in the city of Belém, Pará, Brazil

Cryptococcus neoformans and Cryptococcus gattii are important agents of meningoencephalitis in humans in the city of Belém. This clinical data suggests that the region may be a highly endemic area for the pathogenic Cryptococcus species within the state of Pará (PA), Northern Brazil. Preliminary analysis of 11 environmental samples from the city of Belém showed two positive locations, including a hollow of a kassod tree (Senna siamea) colonized simultaneously by C. gattii molecular type VGII and C. neoformans molecular type VNI, and a birdcage in a commercial aviary positive for C. neoformans, molecular type VNI. This is the first evidence of an environmental occurrence of molecular types VNI and VGII in PA.

Primary endemic Cryptococcosis gattii by molecular type VGII in the state of Pará, Brazil

In order to study the infectious agents causing human disseminated cryptococcosis in the state of Pará, North Brazil, 56 isolates of Cryptococcusspp. (54 isolated from cerebral spinal fluid and two from blood cultures) from 43 cases diagnosed between 2003-2007 were analysed. The species were determined through morphological and physiological tests and genotypes were determined by URA5-RFLP and PCR-fingerprinting (wild-type phage M13). The following species and genotypes were identified: Cryptococcus neoformans VNI (28/56, 50 percent), Cryptococcus gattii VGII (25/56, 44.64 percent) and C. gattii VGI (3/56, 5.26 percent). The genotype VNI occurred in 12 out of 14 HIV-positive adults, whereas the genotype VGII occurred in 11 out of 21 HIV-negative adults (p < 0.02, OR = 6.6 IC95 percent 0.98-56.0). All patients less than 12 years old were HIV negative and six cases were caused by the VGII genotype, one by the VGI and one by VNI. Therefore, endemic primary mycosis in HIV-negative individuals, including an unexpectedly high number of children, caused by the VGII genotype deserves further study and suggests the need for surveillance on cryptococcal infection in the state of Pará, Eastern Amazon.

Regional pattern of the molecular types of Cryptococcus neoformans and Cryptococcus gattii in Brazil

The molecular types of 443 Brazilian isolates of Cryptococcus neoformans and Cryptococcus gattii were analyzed to determine their geographic distribution within Brazil and their underlying host conditions. The following data, imported from previous epidemiological studies as well as two culture collections, were analyzed for: place of isolation, source (clinical or environmental), host risk factors, species, serotype, mating type, and molecular type. Molecular typing by PCR-fingerprinting using primers for the minisatellite-specific core sequence of the wild-type phage M13 or microsatellites [(GACA)4, (GTG)5], restriction fragment length polymorphism of URA5 gene analysis, and/or amplified fragment length polymorphism (AFLP) identified eight major genotypes: VNI/AFLP1, VNII/AFLP1A, VNIII/AFLP2, and VNIV/AFLP3 for C. neoformans, and VGI/AFLP4, VGII/AFLP6, VGIII/AFLP5, and VGIV/AFLP7 for C. gattii. The most common molecular type found in Brazil was VNI (64 percent), followed by VGII (21 percent), VNII (5 percent), VGIII (4 percent), VGI and VNIV (3 percent each), and VNIII (< 1 percent). Primary cryptococcosis caused by the molecular type VGII (serotype B, MAT) prevails in immunocompetent hosts in the North and Northeast regions, disclosing an endemic regional pattern for this specific molecular type in the Northern Brazil.

Diversidade genética e suscetibilidade antifúngica de cepas ambientais e clínicas de Cryptococcus neoformans e Cryptococcus gattii no Brasil / Genetic diversity and antifungal susceptibility of environmental and clinical strains of Cryptococcus neoformans and Cryptococcus gattii in Brazil

A criptococose representa importante problem a de saúde pública no Brasil. É micose de caráter sistêmico, causada por espécies do “complexo Cryptococcus ”: Cryptococcus neoformans (Cn) e Cryptococcus gattii (Cg). Cn é cosmopolita, causando principalmente meningoencefalite em hospedeiros imunodeprimidos. Já Cg causa meningite e pneumonia fúngica em hospedeiros imunocompetentes de ár eas tropicais e subtropicais, com expressiva letalidade no Brasil (56 por cento). Técnicas moleculares permitem identificar espécies e os principais tipos moleculares destes agentes, VNI, VN II, VNIII, VNIV (Cn) e VGI, VGII, VGIII e VGIV (Cg). O tipo VNI é oportunista e predominante em todo mundo, associado a AIDS e outras condições de imunodepressão; VGI é o tipo mo lecular de Cg mais comum no mundo, porém VGII parece ser o mais virulento, sendo o agente da única epidemia de criptococose em humanos e animais já relatada, e que ainda se encontra em evolução no Canadá. O objetivo deste trabalho foi realizar estudo do perfil de sensibilidade antifúngica e genotípico de isolados brasileiros de Cn e Cg. O método de microdiluição em caldo foi utilizado para os testes de suscetibilidade antifúngica de 30 Cn e 57 Cg frente a 9 drogas. A tipagem molecular foi realizada por “PCR fingerprinting” e URA5 -RFLP, o tipo sexuado foi determinado por PCR com primers específicos do gene de ferom ônio, e isolados atípicos foram caracterizados pela técnica de RCA (“Rolling Circle Amplifi cation”). Cg demonstrou ser menos suscetível do que Cn frente a 7 das 9 drogas testadas...