Abstract

P. brasiliensis (Pb) can be isolated frequently from its parasitic yeast form of patients with paracoccidioidomycosis (PCM), naturally infected armadillos and dogs, but the same does not occur for its saprobe condition in nature, where the fungus occur in the mycelial form, producing the infective propagula. Evidences indicate that the fungus grows in soil; however, its isolation from this material is rare and casual. Spider webs are biomaterials with a typical diffuse network and adhesiveness, which may be found frequently at the entrance of armadillo's burrows. The aim of this study was to evaluate the presence of Pb in spider web samples related to the habitat of armadillos, in an area where the fungus was previously isolated from these animals, in Botucatu endemic area of PCM. Spider webs were collected using sterile wooden sticks, packed in test tubes, which were used both for culture in Mycosel Agar (at 35ºC) and also for molecular detection through Nested-PCR with specific primers (ITS rDNA region) for Pb; it was also collected in microscopic slides, which were stained with lacto-phenol cotton blue and microscopically analyzed. It was observed the culture of several dimorphic and/or yeast colonies, which were identified, morphologically and also molecularly by sequencing of ITS-5.8S rDNA, as belonging to the genera Aspergillus, Blastobotrys, Penicillium, Candida, and Sporothrix. Pb was not obtained by culture. The nested-PCR analyses in spider webs showed positive amplification for Pb, with identity confirmed by sequencing of the amplicons, in four samples from a total of 78 evaluated. The microscopic analyses showed different fungal spores adhered to the webs. Spider webs works as a natural air filter and, due to its adhesiveness, represent a simple and effective strategy for capturing aerosols for environmental study of pathogenic and / or opportunistic airborne fungi.਀䘀椀渀愀渀挀椀愀氀 匀甀瀀瀀漀爀琀㨀 䘀愀瀀攀猀瀀 ⴀ ㈀　　㤀⼀㄀㠀㌀㄀㔀ⴀ㈀