Environmental Mapping of Paracoccidioides spp. in Brazil Reveals New Clues into Genetic Diversity, Biogeography and Wild Host Association

PLoS Negl Trop Dis. 2016 Apr 5;10(4):e0004606. doi: 10.1371/journal.pntd.0004606. eCollection 2016 Apr.

Abstract

Background: Paracoccidioides brasiliensis and Paracoccidioides lutzii are the etiological agents of Paracoccidioidomycosis (PCM), and are easily isolated from human patients. However, due to human migration and a long latency period, clinical isolates do not reflect the spatial distribution of these pathogens. Molecular detection of P. brasiliensis and P. lutzii from soil, as well as their isolation from wild animals such as armadillos, are important for monitoring their environmental and geographical distribution. This study aimed to detect and, for the first time, evaluate the genetic diversity of P. brasiliensis and P. lutzii for Paracoccidioidomycosis in endemic and non-endemic areas of the environment, by using Nested PCR and in situ hybridization techniques.

Methods/principal findings: Aerosol (n = 16) and soil (n = 34) samples from armadillo burrows, as well as armadillos (n = 7) were collected in endemic and non-endemic areas of PCM in the Southeastern, Midwestern and Northern regions of Brazil. Both P. brasiliensis and P. lutzii were detected in soil (67.5%) and aerosols (81%) by PCR of Internal Transcribed Spacer (ITS) region (60%), and also by in situ hybridization (83%). Fungal isolation from armadillo tissues was not possible. Sequences from both species of P. brasiliensis and P. lutzii were detected in all regions. In addition, we identified genetic Paracoccidioides variants in soil and aerosol samples which have never been reported before in clinical or armadillo samples, suggesting greater genetic variability in the environment than in vertebrate hosts.

Conclusions/significance: Data may reflect the actual occurrence of Paracoccidioides species in their saprobic habitat, despite their absence/non-detection in seven armadillos evaluated in regions with high prevalence of PCM infection by P. lutzii. These results may indicate a possible ecological difference between P. brasiliensis and P. lutzii concerning their wild hosts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Armadillos / microbiology*
  • Brazil
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • DNA, Ribosomal Spacer / chemistry
  • DNA, Ribosomal Spacer / genetics
  • Environmental Microbiology*
  • Genetic Variation*
  • In Situ Hybridization
  • Paracoccidioides / classification*
  • Paracoccidioides / genetics
  • Paracoccidioides / isolation & purification*
  • Phylogeography*
  • Polymerase Chain Reaction

Substances

  • DNA, Fungal
  • DNA, Ribosomal Spacer

Grants and funding

This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP grant number 2012/03233-3 and 2012/14047-6) and by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.