It is possible that Coccidioides spp., Cryptococcus neoformans and C. gattii, interacting together and/or with other living elements of the soil microbiota, as well as with several other hosts, may generate adaptations and select lineages of these pathogenic fungi.
this website The demonstration of naturally acquired coccidioidomycosis in D. novemcinctus armadillos captured in Piauí reinforces the complexity of this subject [23]. Nevertheless, there have been no investigations of naturally acquired coccidioidomycosis in other species of armadillos, or in other animals such as rodents, foxes, goats, horses, donkeys, cattle and other mammals. Molecular biological techniques have been used to identify pathogenic fungi. Sandhu et al. (1995) analyzed 116 cultures of several human pathogenic fungi using the universal primers U1 and U2 to amplify the conserved 28S rDNA region of fungi, which was then hybridized with probes specific for each fungal species [18]. Sixteen clinical isolates of C. immitis tested by this method demonstrated 100% positivity in identifying this species. Another approach used for the identification of isolates of C. immitis is direct PCR using primers with nucleotide sequences based on the gene csa, which is a 19-kDa specific C. immitis antigen secreted in the growth phase
of fungal cultures that generates a product of about 519 bp [24]. In another study, Bezerra et al. (2006) obtained 100% positivity analyzing the DNA of 19 cultures of C. immitis: twelve clinical isolates from the state of Vorinostat chemical structure Piauí and seven isolates preserved for see more 50-75 years in the culture collection of the Department
of Mycology from the Instituto Oswaldo Cruz at FIOCRUZ in Rio de Janeiro [19]. Regarding the development of molecular methods for the detection of Coccidioides spp. directly in soil samples, obtaining an adequate DNA preparation represented a large challenge. Using mechanical selleck chemicals llc agitation followed by direct cellular enzymatic lysis, we obtained DNA samples with a molecular weight concentrated above 1.5 kb, which were suitable for the amplification reactions by PCR. It should be mentioned that only recently adequate equipment and a Fast DNA SPIN kit for soil (QBIOgene, Carlsbad, CA, USA) allowed the attainment of this suitable DNA from soil samples. In the present study, the primers designed to detect Coccidioides spp. 28S rDNA in soil took into consideration the low number of copies of the target DNA present in soil. This permitted the detection of Coccidioides spp. 28S rDNA in six isolates from the USA and two from Argentina, as well as in thirteen Brazilian isolates. The molecular detection of any of the Coccidioides species in soil or in clinical specimens is of equal importance. Optimization of direct PCR with specific primers to detect C. immitis was first performed with DNA extracted from 21 lineages of Coccidioides spp.