Dárcio Luiz de Sousa Júnior

Objective: The study aimed to optimize protocols for the joint extraction of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from 0.025 × 10⁶ CFU of Candida albicans, targeting to overcome the challenges in the extraction of these genetic materials. Materials and methods: From this, treated silicon carbide (SiC) granules were added to fungal samples from methods 1, 2, and 3 obtained from aliquots of BHI or Sabouraud medium to cause cell lysis and enable the isolation of these macromolecules by phenol and chloroform. The concentration and integrity of the extracted nucleic acids were analyzed, respectively, by spectrophotometry using the A₂₆₀/A₂₈₀ ratios and 1% agarose gel electrophoresis. Results: Therefore, method 3 is the one that most comprises samples considered pure of both DNA and RNA, simultaneously. Furthermore, the presence of intact RNAs corresponding to the base pair size such as 5.8 S rRNA and tRNA was verified during electrophoresis, considering the particularities of RNA, which makes it very unstable and easily degraded. Conclusions: Thus, it results in a faster and simpler method in addition to obtain promising results using minimal amounts of biological sample and offering a valuable alternative for small laboratories to work with molecular biology.

The objective of this study was to evaluate the antifungal activity of eugenol against the genomic DNA of Candida albicans. A quantitative experimental study was conducted with the C. albicans INCQS 40006 strain. Eugenol was tested in concentrations of 12.5%, 25%, 50%, 75% and 100%. The test substance remained in contact with the fungus for 24 hours in a BHI medium in a microbio-logical greenhouse, then the DNA extraction was performed and part of the sample obtained was washed with TE to remove the Eugenol. The fungus was placed in BHI again and placed for 24 hours in a microbiological greenhouse. At the end, the samples with 100% eugenol went through seeds in Sabouraud Dextrose for confirmatory tests. Thus, from the electrophoresis, which admits evaluating the quality of the extracted material, preserving its characteristics and integrity, it was observed in the eugenol concentration test (test 1), in which the eugenol was in direct contact with the pathogen, that in concentrations of equal to or greater than 75% of the test substance the pathogen had no proliferative power. In which the fungus's ability to recover is evaluated (test 2), eugenol was removed from the sample, noting that in 100% of the test substance the fungus did not have the ability to recover, resulting in definitive cell death.

The study aimed to optimize protocols for the joint extraction of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) from 0.025 x 106 CFU of Candida albicans, aiming to overcome challenges in the extraction of these genetic materials. From this, treated Silicon Carbide (SiC) granules were added to fungal samples from methods 1, 2 and 3 obtained from aliquots of BHI or Sabouraud medium to cause cell lysis and enable the isolation of these macromolecules by phenol and chloroform. The concentration and integrity of nucleic acids were obtained, respectively, by spectrophotometry using A260/A280 ratios and 1% agarose gel electrophoresis. Therefore, method 3 is the one that most comprises samples considered pure of both DNA and RNA, simultaneously. Furthermore, the presence of intact RNAs corresponding to the base pair size, such as 5.8 S rRNA and tRNA, was verified during electrophoresis, taking into account the particularities of RNA, which makes it very unstable and easily degraded. Therefore, it results in a faster and simpler method in addition to obtaining promising results using minimal amounts of biological sample, thus offering a valuable alternative for small laboratories working with molecular biology.