Contributions towards the molecular approach of paracoccidoidoimycosis in South America

SOSA MA; GIUSIANO G; ROJAS F

Salvador de Bahia

Congreso; 17th Infocus and 1st ISHAM Latin American Congress; 2019

Paracoccidioides brasiliensis and the related phylogenetic cryptic species included in the brasiliensis clade (P. americana, P. restrepiensis, P. venezuelensis) and P. lutzii are the etiological agents of paracoccidoidoimycosis (PCM), one of the most important systemic mycoses in Latin America. They are dimorphic fungi, with a mycelial life cycle in soil and a yeast phase associated with tissues of mammalian hosts. Molecular epidemiology in South America is not well known; even more, the knowledge of PCM from certain countries is scarce. In order to know the current status of the clinical and epidemiological characteristics of PCM in Argentina, a multicentric study coordinated by Departamento Micología, Instituto de Medicina Regional, Universidad Nacional del Nordeste (Argentina) was started. Subsequently, other South American countries were included in the study. In this work 31 Paracoccidioides isolates from Argentina (15), Paraguay (9), Perú (6) and Bolivia (1) were genotyped by sequencing GP43-exon-2 region and by PCR-RFLP of alpha tubulin (tub1) gene. DNA was extracted from 14-day colonies using CTAB method. TUB1 gene fragments were amplified using primers α-Tub F (5-CTG GGA GGT ATG ATA ACACTG C-3) and α-Tub R (5-CGT CGG GCT ATT CAGATT TAA G-3) described by Roberto et al and Gp43 gene fragments using primers GP43-E2F ( 5-CCAGGAGGCGTGCAGGTGTCCC-3) and Gp43-E2R (5-GCCCCCTCCGTCTTCCATGTCC-3) described by Teixeira et al.P. brasiliensis S1 was the major group recognized followed by PS3. Phylogenomics analysis suggests that two different P. brasiliensis genotypes are endemic in Argentina: P. brasiliensis S1a and S1b. In contrast, paraguayan isolates clustered only with S1b. This results are according with a previous study using the whole genome sequencing typing suggesting multiple P. brasiliensis dispersion events into endemic areas of PCM in Argentina and Paraguay. All peruvian isolates clustered whit P. restrepensis. Even bolivian isolate was the first one identified by molecular methods, considering we studied only one, we cannot make any epidemiological conclusion.Our results confirm the genetic diversity and the geographical relationship in the distribution of the cryptic species of P. brasiliensis.