Virulence profile of ten Paracoccidioides brasiliensis isolates: association with morphologic and genetic patterns.
Ten isolates of Paracoccidioides brasiliensis were examined for differences in virulence in outbred mice intravenously inoculated with the fungus, associated with mycelial morphology, and genetic patterns measured by random amplified polymorphic DNA (RAPD). Virulence was evaluated by viable yeast cell recovery from lungs and demonstration of histopathologic lesions in different organs. The results showed that the isolates presented four virulence degrees: high virulence, intermediate, low and non-virulence. RAPD clustered the isolates studied in two main groups with 56% of genetic similarity. Strains with low virulence, Pb265 or the non-virulent, Pb192, showed glabrous/cerebriform morphology and high genetic similarity (98.7%) when compared to the other isolates studied. The same was observed with Bt79 and Bt83 that shared 96% genetic similarity, cottony colonies and high virulence. The RAPD technique could only discriminate P. brasiliensis isolates according to glabrous/cerebriform or cottony colonies, and also high from low virulence strains. Isolates with intermediate virulence such as Pb18, Pb18B6, Bt32 and Bt56 showed variability in their similarity coefficient suggesting that RAPD was able to detect genetic variability in this fungal species. Virulence profile of P. brasiliensis demonstrated that both mycelial morphologic extreme phenotypes may be associated with fungal virulence and their in vitro subculture time. Thus, RAPD technique analysis employed in association with virulence, morphologic and immunologic aspects might prove adequate to detect differences between P. brasiliensis isolates.
Kurokawa
Cilmery S
CS
Departamento de Pediatria, Faculdade de Medicina, UNESP, Botucatu, São Paulo, SP, Brazil. kurokawa@fmb.unesp.br
Lopes
Catalina R
CR
Sugizaki
Maria F
MF
Kuramae
Eiko E
EE
Franco
Marcello F
MF
Peraçoli
Maria T S
MT
D000818:Animals; D004271:DNA, Fungal / Q000032:analysis; D006801:Humans; D008172:Lung Diseases, Fungal / Q000382:microbiology / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010228:Paracoccidioides / Q000235:genetics / Q000472:pathogenicity*; D010229:Paracoccidioidomycosis / Q000382:microbiology / Q000473:pathology; D010641:Phenotype; D019105:Random Amplified Polymorphic DNA Technique; D014774:Virulence
D004271:DNA, Fungal