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Molecular characterization of Pythium insidiosum by the sequencing of genes coding for Tef-1± and ± and ² tubulin and analysis of the restriction pattern by pulse-field gel electrophoresis (PFGE)

Grant number: 18/24507-0
Support type:Scholarships in Brazil - Master
Effective date (Start): May 01, 2019
Effective date (End): February 29, 2020
Field of knowledge:Biological Sciences - Microbiology - Applied Microbiology
Principal researcher:Sandra de Moraes Gimenes Bosco
Grantee:Ana Carolina do Prado
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil


Pythium insidiosum is the etiological agent of pythiosis, a chronic granulomatous infection, prevalent in tropical and subtropical regions that affect mammals, mainly horses, dogs and humans. This false fungus has a wide geographical distribution being very prevalent in South America (equine and canine pythosis) and in Thailand (human pythosis). Molecular studies have allowed early diagnosis and understanding of phylogenetic relationships on this pathogen. For these purposes some genomic regions have been shown to be efficient targets, such as the cytochrome oxidase (COX II) region and exo-1,3-²-glucanase. Sequence polymorphisms between the COX II and exo-1,3-²-glucanase genes allowed differentiation, at cluster level, between the American and Asian strains, suggesting a possible clonal expansion among the population of P. insidiosum in the Americas, since the greatest genetic diversity is observed in isolates from the Asian region. Other genes, such as translation elongation factor (Tef 1-±) and ± and ²-tubulin were efficiently used in other fungal groups, but were not yet evaluated in P. insidiosum. Genomic studies using the PFGE technique were also not used in the analysis of P. insidiosum populations, and such approach would be useful in the intraspecific comparison. The present project aims to establish phylogenetic relationships among isolates of P. insidiosum by sequencing the gene coding regions for the Tef-1± and ± and ² tubulin regions, in order to identify a new marker to characterize the differences between Brazilian (American) and Thai (Asian), in addition to standardizing the PFGE technique for this species, allowing to estimate the distribution and phylogeography of the clones in both regions. (AU)