Repositorio
Institucional

Santos, Cledir

Rabello, Vanessa Brito de Souza

Correa-Moreira, Danielly

Pinto, Tatiana Casto Abreu

Procopio-Azevedo, Anna Carolina

Boechat, Jessica

Coelho, Rowena Alves

Almeida-Paes, Rodrigo

Costa, Gisela

Lima, Nelson

Zancope-Oliveira, Rosely Maria

Oliveira, Manoel Marques Evangelista

Preservation Methods in Isolates of Sporothrix Characterized by Polyphasic Approach

MDPI

JOURNAL OF FUNGI

en

Sporotrichosis is a subcutaneous mycosis with worldwide distribution and caused by eight pathogenic species of the Sporothrix genus. Different ex situ preservation methods are used around the world to maintain the survival, morphophysiological and genetic traits of fungal strains isolated from patients with sporotrichosis for long terms. The main aim of the present study was to evaluate the survival, phenotypic and genotypic stability of Sporothrix strains after preservation on PDA slant stored at 4 degrees C, sterile water and cryopreservation at -80 degrees C, for a period of time of 6, 12, 18 and 24 months of storage. Eight clinical Sporothrix isolates were identified based on a polyphasic approach consisting of classical macro- and micro-morphological traits, biochemical assays, proteomic profiles by MALDI-TOF MS and molecular biology. According to the final identification, one strain was identified as S. schenckii (CMRVS 40428) and seven strains were re-identified as S. brasiliensis (CMRVS 40421, CMRVS 40423, CMRVS 40424, CMRVS 40425, CMRVS 40426, CMRVS 40427 and CMRVS 40433). In addition, it was observed that the isolates survived after the different time points of storage in distilled water, PDA slant and cryopreservation at -80 degrees C. For fungi preserved in water, low polymorphisms were detected by the partial sequencing of beta-tubulin. Cryopreservation at -80 degrees C induced morphological changes in one single isolate. The proteomic profiles obtained by MALDI-TOF MS after preservation showed differences among the methods. In conclusion, preservation on agar slant stored at 4 degrees C was the most effective method to preserve the eight clinical Sporothrix strains. This method produced less change in the phenotypic traits and kept the genetic integrity of all strains. Agar slant stored at 4 degrees C is a simple and inexpensive method and can be especially used in culture collections of limited funding and resources.

2023

artículo original

10.3390/jof9010034

PDF

preservation

Sporothrix

phenotype

genotype

proteomic

stability

Microbiología

Micología

preservación

Esporotrix

fenotipo

genotipo

proteómico

estabilidad

versión publicada

dorada

verde

acceso abierto

acceso abierto

WOS:000914789700001

artículo original

CC BY 4.0

CC BY 4.0

Microbiología

Micología