Detection of Paracoccidioides spp. in environmental samples and differentiation of the complex P. brasiliensis of the P. lutzii species by Nested PCR and in situ hybridization.

• Grant number: 12/03233-3
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: June 01, 2012
• End date: February 28, 2015
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Eduardo Bagagli
• Grantee: Thales Domingos Arantes
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The soil is the probable habitat of the pathogenic fungus Paracoccidioides brasiliensis, due to its molecular detection in such environmental sample, which may be associated to the high frequency of infection of rural workers as well as wild animals (Dasypus novencinctus, Cabassous centralis and Choloepus didactylus). In the last few years, the molecular detection showed to be promising for the ecological studies about this pathogen, mainly by PCR and Nested PCR techniques. This project aims to detect and differentiate the species Paracoccidioides brasiliensis (complex S1, PS2 and PS3) and Paracoccidioides lutzii in the environment by performing Nested PCR and TSA-FISH, respectively, in aerosol and soil samples from armadillo's burrows from endemic areas for Paracoccidioidomycosis in the Southeast and Mid-Western regions of Brazil. The hybridization with oligonucleotides probes, known as FISH (Fluorescence in situ Hybridization) can be associated with the TSA (Tyramide Signal Amplification) technique, which amplifies the signal emitted by the fluorescent probe. Besides the environmental detection of Paracoccidioides sp. by FISH-TSA, the occurrence of armadillos infected with P. lutzii in the Brazilian Mid-Western, where this species is prevalent, will be studied for the first time (no armadillo from the area has been evaluated so far). Since the current geographic distribution of isolates from Paracoccidioides genus is based mainly on clinical samples and therefore presents the interference of host migration, we believe that the approach here proposed shows a great potential for a better understanding of the biogeography of Paracoccidioides sp., given that the fungal detection will be carried out directly from the environment.

Scientific publications

(4)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

ARANTES, THALES DOMINGOS; THEODORO, RAQUEL CORDEIRO; TEIXEIRA, MARCUS DE MELO; BAGAGLI, EDUARDO. Use of fluorescent oligonucleotide probes for differentiation between Paracoccidioides brasiliensis and Paracoccidioides lutzii in yeast and mycelial phase. Memórias do Instituto Oswaldo Cruz, v. 112, n. 2, p. 140+, FEB 2017. (12/03233-3, 12/14047-6)

ARANTES, THALES DOMINGOS; THEODORO, RAQUEL CORDEIRO; TEIXEIRA, MARCUS DE MELO; GIMENES BOSCO, SANDRA DE MORAES; BAGAGLI, EDUARDO. Environmental Mapping of Paracoccidioides spp. in Brazil Reveals New Clues into Genetic Diversity, Biogeography and Wild Host Association. PLoS Neglected Tropical Diseases, v. 10, n. 4, APR 2016. (12/14047-6, 12/03233-3)

ARANTES, THALES DOMINGOS; BAGAGLI, EDUARDO; NINO-VEGA, GUSTAVO; SAN-BLAS, GIOCONDA; THEODORO, RAQUEL CORDEIRO. Paracoccidioides brasiliensis AND Paracoccidioides lutzii, A SECRET LOVE AFFAIR. Revista do Instituto de Medicina Tropical de São Paulo, v. 57, n. 19, p. 25-30, SEP 2015. (12/14047-6, 12/03233-3)

ARANTES, THALES DOMINGOS; THEODORO, RAQUEL CORDEIRO; TEIXEIRA, MARCUS DE MELO; BAGAGLI, EDUARDO. Use of fluorescent oligonucleotide probes for differentiation between Paracoccidioides brasiliensis and Paracoccidioides lutzii in yeast and mycelial phase. Memórias do Instituto Oswaldo Cruz, v. 112, n. 2, p. 8-pg., 2017-02-01. (12/14047-6, 12/03233-3)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

ARANTES, Thales Domingos. Detecção do Paracoccidioides spp. em amostras ambientais e diferenciação do complexo P. brasiliensis da espécie P. lutzii por Nested PCR e hibridização in situ. 2015. Doctoral Thesis - Universidade Estadual Paulista (Unesp). Instituto de Biociências. Botucatu Botucatu.