Lippia origanoides Essential Oil or Thymol in Combination with Fluconazole Produces Damage to Cells and Reverses the Azole-Resistant Phenotype of a Candida tropicalis Strain
Abstract
:1. Introduction
2. Materials and Methods
2.1. Compounds
2.2. Minimum Inhibitory Concentration
2.3. Interaction Assay
2.4. Assessment of Plasmatic Membrane Integrity
2.5. Evaluation of Mitochondrial Membrane Potential (∆ψm)
2.6. Mitochondrial Superoxide Indicator
2.7. Nuclear Effect Assessment
2.8. Cell Cycle Assessment
2.9. Morphology Assessment by Scanning Electron Microscopy
2.10. Ultrastructure Assessment by Transmission Electron Microscopy
3. Results
3.1. Lippia Origanoides EO or Thymol in Combination with FLC Reduce Mitochondrial Membrane Potential (∆ψm) but Do Not Affect the Plasmatic Membrane
3.2. Thymol Used Alone and in Combination with FLC Increased Mitochondrial Superoxide Production
3.3. Lippia Origanoides EO or Thymol in Combination with FLC Causes Nuclear Alterations
3.4. Lippia origanoides EO or Thymol in Combination with FLC Causes Alterations in the Cell Cycle
3.5. Lippia origanoides EO or Thymol in Combination with FLC Cause Both Superficial and Ultrastructural Alterations
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Plant Species and Chemotype | Collection Site | Voucher Number | Principal Compounds (Relative Amount, %) |
---|---|---|---|
Lippia origanoides (Thymol + p-cymene) chemotype | Bucaramanga—Santander, Colombia | UIS * herbarium 22039 | Thymol (49.4), p-cymene (19.1), γ-terpinene (9.2), β-myrcene (5.2), α-terpinene (2.9), carvacrol (2.7), thymyl methyl ether (1.8), trans-β-caryophyllene (1.6), cis-β-ocimene (1.2), and limonene (0.9). |
Combination | FLC MIC (X) | Thymol or EO MIC (X) |
---|---|---|
1 | 0 | 8 |
2 | 0.5 | 4 |
3 | 1 | 2 |
4 | 2 | 1 |
5 | 4 | 0.5 |
6 | 8 | 0 |
Strain | Compound/ Combination | GM-MIC (µg/mL) | FICI | Interaction Type | No. Times ↓ MIC | ||
---|---|---|---|---|---|---|---|
FLC | EO | Thymol | |||||
C. tropicalisATCC 200956 * | FLC | >64 | - | - | - | - | - |
EO | 128 | - | - | - | - | - | |
Thymol | 128 | - | - | - | - | - | |
EO/FLC | 32/2 | 0.28 | Synergistic | 32 | 4 | - | |
Thymol/FLC | 32/2 | 0.28 | Synergistic | 32 | - | 4 | |
C. tropicalisATCC 750 ** | FLC | 2 | - | - | - | - | - |
EO | 512 | - | - | - | - | - | |
Thymol | 256 | - | - | - | - | - | |
EO/FLC | 181/0.25 | 0.53 | Additive | 8.0 | 2.8 | - | |
Thymol/FLC | 42/0.60 | 0.47 | Synergistic | 3.3 | - | 6 |
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Zapata-Zapata, C.; Rojas-López, M.; García, L.T.; Quintero, W.; Terrón, M.C.; Luque, D.; Mesa-Arango, A.C. Lippia origanoides Essential Oil or Thymol in Combination with Fluconazole Produces Damage to Cells and Reverses the Azole-Resistant Phenotype of a Candida tropicalis Strain. J. Fungi 2023, 9, 888. https://doi.org/10.3390/jof9090888
Zapata-Zapata C, Rojas-López M, García LT, Quintero W, Terrón MC, Luque D, Mesa-Arango AC. Lippia origanoides Essential Oil or Thymol in Combination with Fluconazole Produces Damage to Cells and Reverses the Azole-Resistant Phenotype of a Candida tropicalis Strain. Journal of Fungi. 2023; 9(9):888. https://doi.org/10.3390/jof9090888
Chicago/Turabian StyleZapata-Zapata, Carolina, Mauricio Rojas-López, Liliana T. García, Wendy Quintero, María C. Terrón, Daniel Luque, and Ana C. Mesa-Arango. 2023. "Lippia origanoides Essential Oil or Thymol in Combination with Fluconazole Produces Damage to Cells and Reverses the Azole-Resistant Phenotype of a Candida tropicalis Strain" Journal of Fungi 9, no. 9: 888. https://doi.org/10.3390/jof9090888