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Preservation Methods in Isolates of Sporothrix Characterized by Polyphasic Approach
Vanessa Brito de Souza Rabello1, Danielly Corrêa-Moreira2,3, Cledir Santos4
1Laboratory of Mycology, Evandro Chagas National Institute of Infectious Diseases, Fiocruz. Av. Brasil, 4365, Manguinhos, Rio de Janeiro 21040-360, Brazil.
Journal of Fungi (Basel, Switzerland)
|January 21, 2023
Summary
Preserving Sporothrix fungal strains on PDA slant at 4°C is most effective for maintaining survival, phenotypic traits, and genetic integrity. This simple, inexpensive method is ideal for resource-limited culture collections.
Area of Science:
- Medical Mycology
- Fungal Strain Preservation
- Molecular Biology
Background:
- Sporotrichosis is a globally distributed subcutaneous mycosis caused by *Sporothrix* species.
- Effective ex situ preservation methods are crucial for maintaining fungal strain viability, morphophysiology, and genetic traits.
- Various preservation techniques exist, but their long-term efficacy for *Sporothrix* requires evaluation.
Purpose of the Study:
- To assess the survival, phenotypic stability, and genotypic stability of *Sporothrix* strains under different preservation methods.
- To compare the effectiveness of PDA slant storage at 4°C, sterile water, and cryopreservation at -80°C over 24 months.
- To identify the optimal preservation method for clinical *Sporothrix* isolates.
Main Methods:
- Eight clinical *Sporothrix* isolates (*S. schenckii* and *S. brasiliensis*) were preserved using three methods: PDA slant at 4°C, sterile water, and cryopreservation at -80°C.
- Samples were evaluated at 6, 12, 18, and 24 months.
- Methods included macro- and micro-morphological analysis, biochemical assays, MALDI-TOF MS proteomic profiling, and partial β-tubulin gene sequencing.
Main Results:
- All tested preservation methods allowed for *Sporothrix* strain survival over 24 months.
- Preservation in sterile water showed low polymorphisms in β-tubulin sequencing.
- Cryopreservation at -80°C induced morphological changes in one isolate, and MALDI-TOF MS revealed differences in proteomic profiles among preservation methods.
- Preservation on PDA slant at 4°C resulted in minimal phenotypic changes and maintained genetic integrity.
Conclusions:
- Preservation on PDA slant at 4°C is the most effective method for long-term preservation of clinical *Sporothrix* strains.
- This method ensures fungal survival, preserves phenotypic characteristics, and maintains genetic stability.
- The simplicity and low cost of PDA slant storage make it highly suitable for culture collections with limited resources.
