Limonin Enhances the Antifungal Activity of Eugenol Nanoemulsion against Penicillium Italicum In Vitro and In Vivo Tests
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Nanoemulsion Preparation
2.3. Characterization of Nanoemulsions
2.3.1. Particle Size Determination
2.3.2. Optical Microscopy and Confocal Laser Scanning Microscopy
2.4. Antifungal Activity of the Nanoemulsions
2.4.1. Fungal Strains
2.4.2. Determination of MIC and MFC
2.4.3. Influence of the Emulsions on Conidial Germination of P. italicum
2.4.4. Inhibitory Effect of the Nanoemulsion on Mycelial Growth of P. italicum
2.5. Morphological Observation
2.6. Determination of Microbial Cell Membrane Permeability
2.6.1. Extracellular Electric Conductivity Measurement
2.6.2. Leakage of Protein and Nucleic Acid
2.7. Lipid Peroxidation Assay
2.8. Challenge Test of Citrus Fruits
2.9. Statistical Analyses
3. Results
3.1. Characterization of Nanoemulsions
3.2. Antifungal Activity
3.2.1. MICs and MFCs Determination
3.2.2. Inhibition of Spore Germination
3.2.3. Inhibition of Mycelial Growth
3.3. Micromorphological Analysis by SEM
3.4. Influence of Nanoemulsions on the Cell Membrane Permeability
3.4.1. Extracellular Conductivity
3.4.2. Leakage of Nucleic Acid and Protein
3.5. Lipid Peroxidation
3.6. Challenge Test
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Groups | Concentrations a | Mycelial Growth in PDB (3rd Day) | Mycelial Growth on PDA (6th Day) | MIC a | MFC a |
---|---|---|---|---|---|
EG | 1280 | − | − | 320 | 320 |
640 | − | − | |||
320 | − | − | |||
160 | + | ||||
80 | + | ||||
EGL | 1280 | − | − | 160 | 320 |
640 | − | − | |||
320 | − | − | |||
160 | − | + | |||
80 | + | ||||
PDB + Stain | 0 | + | |||
PDB | 0 | − |
Groups | Infected Oranges (%) | Infected Cuts (%) | ||
---|---|---|---|---|
3 d | 5 d | 3 d | 5 d | |
CK | 75.0 ± 12.5 a | 100.0 ± 0.0 a | 59.7 ± 6.4 a | 84.7 ± 6.4 a |
EG | 37.5 ± 12.5 b | 58.3 ± 7.2 b | 29.2 ± 11.0 b | 38.9 ± 8.7 b |
EGL | 20.8 ± 7.2 b | 29.2 ± 7.2 c | 12.5 ± 4.2 c | 19.4 ± 6.4 c |
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Li, Y.; Zhao, R.; Li, Y.; Zhou, Z. Limonin Enhances the Antifungal Activity of Eugenol Nanoemulsion against Penicillium Italicum In Vitro and In Vivo Tests. Microorganisms 2021, 9, 969. https://doi.org/10.3390/microorganisms9050969
Li Y, Zhao R, Li Y, Zhou Z. Limonin Enhances the Antifungal Activity of Eugenol Nanoemulsion against Penicillium Italicum In Vitro and In Vivo Tests. Microorganisms. 2021; 9(5):969. https://doi.org/10.3390/microorganisms9050969
Chicago/Turabian StyleLi, Yi, Runan Zhao, Yan Li, and Zhiqin Zhou. 2021. "Limonin Enhances the Antifungal Activity of Eugenol Nanoemulsion against Penicillium Italicum In Vitro and In Vivo Tests" Microorganisms 9, no. 5: 969. https://doi.org/10.3390/microorganisms9050969