RESUMO
Cryptococcosis is a systemic fungal disease acquired from contaminated environments with propagules of the basidiomycetous yeasts of the Cryptococcus neoformans and C. gattii species complexes. The C. neoformans species complex classically comprises four major molecular types (VNI, VNII, VNIII, and VNIV), and the C. gattii species complex comprises another four (VGI, VGII, VGIII, and VGIV) and the newly identified molecular type VGV. These major molecular types differ in their epidemiological and ecological features, clinical presentations, and therapeutic outcomes. Generally, the most common isolated types are VNI, VGI, and VGII. The epidemiological profile of cryptococcosis in domestic cats is poorly studied and cats can be the sentinels for human infections. Therefore, the present study aimed to determine the molecular characterization of Cryptococcus spp. isolated from domestic cats and their dwellings in the metropolitan area of Rio de Janeiro, Brazil. A total of 36 Cryptococcus spp. strains, both clinical and environmental, from 19 cats were subtyped using multilocus sequence typing (MLST). The ploidy was identified using flow cytometry and the mating type was determined through amplification with specific pheromone primers. All strains were mating type alpha and 6/36 were diploid (all VNII). Most isolates (63.88%) were identified as VNII, a rare molecular type, leading to the consideration that this genotype is more likely related to skin lesions, since there was a high percentage (68.75%) of cats with skin lesions, which is also considered rare. Further studies regarding the molecular epidemiology of cryptococcosis in felines are still needed to clarify the reason for the large proportion of the rare molecular type VNII causing infections in cats.
RESUMO
Two cats infected by C. gattii, presented lesions on the nasal region and respiratory signs. Strains were typed as molecular type VGII, mating type alpha, MLST subtypes ST442 and ST185. Since Rio de Janeiro is known as an endemic area for C. neoformans VNI, these cases might be a warning for a possible emergence of C. gattii VGII in southeast Brazil.
Assuntos
Complicações Infecciosas na Gravidez/microbiologia , Adolescente , Adulto , Animais , Mordeduras e Picadas/complicações , Brasil , Gatos , Feminino , Humanos , Hipertermia Induzida , Gravidez , Complicações Infecciosas na Gravidez/terapia , Sporothrix , Esporotricose/microbiologia , Esporotricose/terapia , Adulto JovemRESUMO
Sporotrichosis is a subcutaneous mycosis diagnosed by isolation of the fungus in culture. Serological tests for help in diagnosis in general do not use purified or recombinant antigens, because there is a paucity of described immunoreactive proteins, especially for the new described Sporothrix species, such as Sporothrix brasiliensis. This study aims to characterise antigens from S. brasiliensis and verify their application in serodiagnosis of sporotrichosis. An immunoblot assay allied with computer-based analysis was used to identify putative antigenic molecules in a cell-free extracts of both morphological phases of this fungus, and to delineate antigenic polymorphism among seven S. brasiliensis isolates and one S. schenckii Brazilian strain. The mycelial and yeast phase of the fungus originated 14 and 23 reactive bands, respectively, which were variable in intensity. An 85 kDa antigen, verified in the yeast phase of the fungus, was observed in all strains used and the immunodominant protein was identified. This protein, however, cross-react with serum samples from patients infected with other pathogens. The results show that the S. brasiliensis cell-free antigen extract is a single and inexpensive source of antigens, and can be applied on the sporotrichosis serodiagnosis.
Assuntos
Antígenos de Fungos/análise , Immunoblotting/métodos , Sporothrix/isolamento & purificação , Esporotricose/microbiologia , Variação Antigênica , Antígenos de Fungos/imunologia , Brasil , Humanos , Sporothrix/crescimento & desenvolvimento , Sporothrix/imunologia , Esporotricose/diagnósticoRESUMO
BACKGROUND: Sporotrichosis is the most common subcutaneous mycosis in Rio de Janeiro. Histopathological examination reveals diffuse granulomatous and suppurative dermatitis, and the fungus is rarely identifiable in tissue. We describe the histopathological features of cutaneous sporotrichosis, and investigate the association between them and the lack of visualization of the fungus. METHODS: A total of 119 samples of confirmed sporotrichosis cases were studied. The characteristics of the inflammatory infiltrate, the presence of epidermal changes, necrosis and fibrosis, and the parasite burden were analyzed. The association between histopathological changes and the lack of visualization of the fungus was evaluated using prevalence ratios. RESULTS: Granulomas were observed in all samples, suppurative granulomas in 100 (84%) and diffuse dermatitis in 114 (95.8%). Liquefaction and caseous necrosis were present in 78 (65.5%) and 52 (43.7%) samples, respectively. The fungus was not seen in 77 (64.7%) samples. Epithelioid, tuberculoid or foreign-body-type granulomas, caseous, fibrinoid or absent necrosis, predominance of lymphocytes among nonphagocytic cells and fibrosis were associated with the lack of visualization of the fungus. CONCLUSION: The histopathological characteristics of sporotrichosis were variable when the causative fungus was not seen. In a proper context, the diagnosis of sporotrichosis remains a potential consideration even in the absence of demonstrable yeast.
Assuntos
Esporotricose/microbiologia , Esporotricose/patologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil , Criança , Feminino , Granuloma/microbiologia , Humanos , Masculino , Pessoa de Meia-Idade , Necrose , Pele/microbiologia , Pele/patologia , Adulto JovemRESUMO
An epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus Sporothrix schenckii, is ongoing in Rio de Janeiro, Brazil, in which cases of human infection are related to exposure to cats. In an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by DNA-based typing methods 19 human and 25 animal S. schenckii isolates from the epidemic, as well as two control strains. To analyse the isolates, the random amplified polymorphic DNA (RAPD) technique was performed using three different primers, together with DNA fingerprinting using the minisatellite derived from the wild-type phage M13 core-sequence. The analyses generated amplicons with considerable polymorphism. Although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. The RAPD profiles of epidemic S. schenckii isolates could be distinguished from that of the United States isolate, displaying 20% similarity to each primer and 60% when amplified with the M13 primer. DNA fingerprinting of S. schenckii isolated from the nails (42.8%) and the oral cavities (66%) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. It is clear that cats act as a vehicle for dissemination of S. schenckii.
Assuntos
Doenças do Gato/microbiologia , Surtos de Doenças , Sporothrix/isolamento & purificação , Esporotricose/microbiologia , Animais , Brasil/epidemiologia , Doenças do Gato/diagnóstico , Doenças do Gato/epidemiologia , Gatos , Impressões Digitais de DNA , DNA Fúngico , Humanos , Técnicas de Tipagem Micológica , Polimorfismo Genético , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sporothrix/genética , Esporotricose/diagnóstico , Esporotricose/epidemiologia , Esporotricose/veterináriaRESUMO
An epidemic of sporotrichosis, a subcutaneous mycosis caused by the fungus Sporothrix schenckii, is ongoing in Rio de Janeiro, Brazil, in which cases of human infection are related to exposure to cats. In an attempt to demonstrate the zoonotic character of this epidemic using molecular methodology, we characterised by DNA-based typing methods 19 human and 25 animal S. schenckii isolates from the epidemic, as well as two control strains. To analyse the isolates, the random amplified polymorphic DNA (RAPD) technique was performed using three different primers, together with DNA fingerprinting using the minisatellite derived from the wild-type phage M13 core-sequence. The analyses generated amplicons with considerable polymorphism. Although isolates exhibited high levels of genetic relatedness, they could be clustered into 5-10 genotypes. The RAPD profiles of epidemic S. schenckii isolates could be distinguished from that of the United States isolate, displaying 20 percent similarity to each primer and 60 percent when amplified with the M13 primer. DNA fingerprinting of S. schenckii isolated from the nails (42.8 percent) and the oral cavities (66 percent) of cats were identical to related human samples, suggesting that there is a common infection source for animals and humans in this epidemic. It is clear that cats act as a vehicle for dissemination of S. schenckii.
Assuntos
Animais , Gatos , Humanos , Doenças do Gato/microbiologia , Surtos de Doenças , Sporothrix/isolamento & purificação , Esporotricose/microbiologia , Brasil/epidemiologia , Doenças do Gato/diagnóstico , Doenças do Gato/epidemiologia , Impressões Digitais de DNA , DNA Fúngico , Técnicas de Tipagem Micológica , Polimorfismo Genético , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sporothrix/genética , Esporotricose/diagnóstico , Esporotricose/epidemiologia , Esporotricose/veterináriaRESUMO
Seventy-four dogs from the State of Rio de Janeiro with ulcerated cutaneous lesions were submitted to clinical, dermatological, parasitological, mycological, histopathological and cytopathological exams, a leishmanin skin test, an indirect immunofluorescence (IIF) test for leishmaniosis, and nonspecific laboratory tests such as blood count and serum biochemistry. Sporothrix schenckii was isolated from 41 dogs and Leishmania (Viannia) braziliensis was isolated from 33 animals. Most dogs with sporotrichosis were from the municipality of Rio de Janeiro (53.7%) and presented ulcerated cutaneous lesions on the head (68.3%). Laboratory alterations in these animals included anemia (58.5%), hypoalbuminemia (83%) and hyperglobulinemia (75.6%). Histopathology revealed the predominance of a chronic granulomatous inflammatory infiltrate (70.7%), and yeast-like structures were detected in 17% of the histopathological exams and in 32% of the cytological exams. Three of 41 dogs with sporotrichosis were seropositive by IIF for leishmaniosis and 2 of 20 animals tested within this group had a positive leishmanin skin test. Similarly, most of the 33 dogs with leishmaniosis were from the municipality of Rio de Janeiro (69.7%) and had ulcerated cutaneous lesions on the head (84.8%). Laboratory alterations in these animals included anemia (66.7%), hypoalbuminemia (100%) and hyperglobulinemia (91%). Histopathology showed the predominance of a chronic granulomatous inflammatory infiltrate (63.6%) and amastigote forms were detected in 30.3% of the histopathological exams and in 31.8% of the 22 cytological exams performed. About 72.7% of the dogs were seropositive by IIF and five of seven animals had a positive skin test. Due to the clinical similarities, histopathological and nonspecific laboratory results similarities, the serological and skin tests for leishmaniosis positive in dogs with sporotrichosis, and the overlapping endemic areas in Rio de Janeiro, the differential diagnosis between the two diseases requires the demonstration of their respective etiological agents.
Assuntos
Doenças do Cão/diagnóstico , Leishmaniose Cutânea/veterinária , Sporothrix/isolamento & purificação , Esporotricose/veterinária , Anemia/diagnóstico , Anemia/epidemiologia , Anemia/veterinária , Animais , Análise Química do Sangue/veterinária , Brasil/epidemiologia , Diagnóstico Diferencial , Doenças do Cão/epidemiologia , Cães , Feminino , Histocitoquímica , Hipergamaglobulinemia/diagnóstico , Hipergamaglobulinemia/epidemiologia , Hipergamaglobulinemia/veterinária , Hipoalbuminemia/diagnóstico , Hipoalbuminemia/epidemiologia , Hipoalbuminemia/veterinária , Leishmania braziliensis/imunologia , Leishmania braziliensis/isolamento & purificação , Leishmaniose Cutânea/diagnóstico , Leishmaniose Cutânea/epidemiologia , Masculino , Estudos Soroepidemiológicos , Pele/microbiologia , Pele/parasitologia , Pele/patologia , Úlcera Cutânea/microbiologia , Úlcera Cutânea/parasitologia , Úlcera Cutânea/patologia , Úlcera Cutânea/veterinária , Sporothrix/imunologia , Esporotricose/diagnóstico , Esporotricose/epidemiologiaRESUMO
In this retrospective study we analyzed 70 HIV patients with cryptococcosis over a 16-years period. The specimens with the best positivity were CSF (97.8%) followed by the culture of urine sediment (86.7%) and blood culture (58.8%). We conclude that the urine could be a useful tool for the diagnosis of cryptococcosis.