RESUMO
The biosynthesis of chondroitinase and hyaluronidase by different isolates of Paracoccidioides brasiliensis was investigated in 20 strains isolated from patients (17 strains), a penguin (Pygocelis adeliae, one strain), an armadillo (Dasypus novemcinctus, one strain) and the environment (dog food, one strain). All the P. brasiliensis isolates studied had the ability to produce chondroitinase and hyaluronidase, although differences in colony morphology and enzyme production were detected among them. These results suggest that further investigations should be carried out in the clinical field in order to clarify the potential role of P. brasiliensis enzyme production in the pathogenesis of paracoccidioidomycosis.
Assuntos
Condroitina ABC Liase/biossíntese , Hialuronoglucosaminidase/biossíntese , Paracoccidioides/enzimologia , Ração Animal/microbiologia , Animais , Tatus , Aves , Humanos , Paracoccidioides/patogenicidade , Paracoccidioidomicose/enzimologia , Paracoccidioidomicose/microbiologia , VirulênciaRESUMO
We have investigated the production of proteinase and phospholipase by 20 different isolates of Paracoccidioides brasiliensis. Isolates were grown in Bacto-peptone, Dextrose, pH 5.5, agar slants, at 27 degrees C for 30 days, and cultures were transferred onto Petri dishes containing basis medium and bovine serum albumin fraction V and sterile egg yolk as substrates for enzyme production, and incubated at 27 degrees C. After 30 days net enzyme activity was visualized and quantitatively evaluated, measuring a ratio between colony diameter and diameter of the transparent (proteinase) or white (phospholipase) ring zone surrounding it. Results demonstrated that all isolates had the ability to produce proteinase and phospholipase, even though variability in enzyme production was noted among different isolates of P. brasiliensis.
Assuntos
Endopeptidases/metabolismo , Paracoccidioides/enzimologia , Fosfolipases/metabolismo , Humanos , Paracoccidioides/citologia , Paracoccidioidomicose/microbiologia , Especificidade da EspécieRESUMO
Sera from two patients with chronic active paracoccidioidomycosis yielded negative double immunodiffusion results with a culture filtrate antigen from Paracoccidioides brasiliensis routinely used in our laboratory. Complement fixation tests were positive for both sera using a polysaccharide-rich antigen. This study reports the results of a more extensive serological investigation of these two sera. Both a somatic antigen and a saline extract from the fungus yielded positive results in the double immunodiffusion. However, the immunodominant 43 kDa glycoprotein antigen showed negative results, although it was recognized by both sera in the Western blot assay. The value of the double immunodiffusion as a single serological test in paracoccidioidomycosis diagnosis is discussed.
Assuntos
Antígenos de Fungos/imunologia , Paracoccidioidomicose/imunologia , Adulto , Idoso , Reações Falso-Negativas , Humanos , Imunodifusão , Masculino , Paracoccidioidomicose/sangueRESUMO
A sample of P. brasiliensis isolated from the spleen and the liver of an armadillo (Dasipus novencinctus) has been analysed under a mycological and immunochemical viewpoint. The armadillo was captured in an area of Tucuruí (State of Pará, Brazil), the animal being already established as an enzootic reservoir of P. brasiliensis at that region of the country. This sample maintained in the fungal collection of the Tropical Medicine Institute of São Paulo (Brazil) numbered 135, has got all the characteristics of P. brasiliensis, with a strong antigenic power and low virulence for guinea-pigs and Wistar rats. The specific exoantigen of P. brasiliensis--the glycoprotein with a molecular weight of 43 kDa--was easily demonstrated with double immunodiffusion, immunoelectrophoresis, SDS-PAGE and immunobloting techniques.
Assuntos
Tatus/microbiologia , Paracoccidioides/isolamento & purificação , Animais , Antígenos de Fungos/isolamento & purificação , Cricetinae , Reservatórios de Doenças , Cobaias , Imunoquímica , Imunoglobulina G , Masculino , Paracoccidioides/imunologia , Ratos , Ratos WistarRESUMO
The Authors show the results obtained through the study of a Paracoccidioides strain isolated from a penguin in the Uruguaian Antartide by GEZUELE et al. (1989). From the fecal mater it was isolated a fungus which was recently considered as a new species of the genus Paracoccidioides--P. antarcticus. However, the mycological and immunochemical studies including the demonstration of the 43 kDa glycoprotein by immunodiffusion test, SDS-PAGE and immunoelectrophoresis disclosed that such strain is similar to P. brasiliensis. Other studies, based on molecular taxonomy, including karyotyping, are the only tools to confirm the possibility of such strain to be a variant of P. brasiliensis. The Authors report the epidemiological significance of that finding and suggest a review in the knowledge of the ecological "niche" of P. brasiliensis.
Assuntos
Aves , Fezes/microbiologia , Paracoccidioides/isolamento & purificação , Animais , Imunoquímica , Imunodifusão , Imunoeletroforese , Masculino , Técnicas de Tipagem Micológica , Paracoccidioides/classificação , Paracoccidioides/imunologia , Testículo/patologiaRESUMO
Antigens from three strains of Aspergillus fumigatus (354, 356, and JIG) and an antiserum against the mixing of these antigens have been produced, and evaluated immunochemically. The antigens were obtained through a modified Coleman & Kaufman technique (culture filtrate concentrated by acetone). Analysis by the immunodiffusion test (ID) against homologous serum has yielded 100% sensitivity (with the studied sera). Concerning heterologous sera we found reactivity with a serum of a patient of candidiasis and another with histoplasmosis. The same result was obtained with a reference antigen in immunodiffusion, showing similar standards of response. Titration of the antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique) a title of 1:128. Using immunoelectrophoresis (IEF), the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one). In SDS-PAGE at 12.5% the antigen has presented a rather complex electrophoretic profile (26 proteic subunits with a molecular weight ranging from 18 a > 100 kDa). Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immunoblotting against the antiserum was carried out.
Assuntos
Antígenos de Fungos/isolamento & purificação , Aspergillus fumigatus/imunologia , Animais , Antígenos de Fungos/análise , Aspergilose/diagnóstico , Humanos , Soros Imunes/análise , Soros Imunes/isolamento & purificação , Imunização , Imunoquímica , Técnicas Imunológicas , Masculino , CoelhosRESUMO
The purpose of this work is obtaining exocellular antigens H and M from 4 H. capsulatum strains using NGTA medium (neopeptone, glucose, thiamine and asparagine) for periods of 1, 2 and 3 months, at 36 degrees C and continuously shaken. The exocellular antigens were evaluated by double immunodiffusion test against H. capsulatum rabbit antiserum, 7 histoplasmosis sera, 4 paracoccidioidomycosis sera and a reference antigen and antibody furnished by C.D.C. (Atlanta--USA). Except for the exocellular antigen from strain B.679 with 1 month of culture, all exocellular antigens obtained from the strains B.679, 58 and O187 showed the H and M bands. The A.811 strain demonstrated only the fraction H. All the exocellular antigens reacted positively with sera from histoplasmosis patients, except those obtained from strains 58 and B.679 with 1 month of culture. With regard to paracoccidioidomycosis patients sera, the exocellular antigens from strains 58 and O187 did not cross-react with them.
Assuntos
Antígenos de Fungos/biossíntese , Histoplasma/imunologia , Meios de Cultura , Imunodifusão/métodosRESUMO
We report the isolation of a strain of Paracoccidioides brasiliensis from a dogfood, probably contaminated with soil, in a Brazilian city. The fungus was isolated on appropriate culture media, and when inoculated into a guinea-pig testis produced orchitis with abundant fungal elements. Histopathology of sections of the testicle showed an inflammatory reaction with P. brasiliensis inside monocytes. Immunological identification confirmed the identity of the isolate.