In Vitro Anti-Leptospiral Activity of Phyllanthus amarus Extracts and Their Combinations with Antibiotics
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sampling and Plant Extraction
2.2. Ellinghausen McCullough Johnson Harris Broth Preparation
2.3. Antibiotic Solution Preparation
2.4. Leptospira Inoculum Preparation
2.5. Antimicrobial Susceptibility Testing
2.6. Synergy Testing by Checkerboard Assay
2.7. Scanning Electron Microscope Analysis
2.8. Statistical Analysis
3. Results
3.1. Antimicrobial Susceptibility Testing
3.2. Synergy Testing
3.3. SEM Analysis
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Leptospira Serovars | AE (µg/mL) | ME (µg/mL) | ||
---|---|---|---|---|
MIC | MBC | MIC | MBC | |
L. interrogans serovar Australis | 400 ± 0.0 | 400 ± 0.0 | 800 ± 0.0 | 800 ± 0.0 |
L. interrogans serovar Bataviae | 100 ± 0.0 | 100 ± 0.0 | 400 ± 0.0 | 400 ± 0.0 |
L. interrogans serovar Caniola | 100 ± 0.0 | 100 ± 0.0 | 400 ± 0.0 | 400 ± 0.0 |
L. interrogans serovar Javanica | 200 ± 0.0 | 200 ± 0.0 | 800 ± 0.0 | 800 ± 0.0 |
Leptospira Serovars | Doxycycline (µg/mL) | Penicillin G (µg/mL) | Ceftriaxone (µg/mL) | |||
---|---|---|---|---|---|---|
MIC | MBC | MIC | MBC | MIC | MBC | |
L. interrogans serovar Australis | 0.78 ± 0.00 | 12.50 ± 0.00 | <0.01 ± 0.00 | <0.01 ± 0.00 | 0.20 ± 0.00 | 0.39 ± 0.00 |
L. interrogans serovar Bataviae | 0.39 ± 0.00 | 12.50 ± 0.00 | <0.01 ± 0.00 | 0.05 ± 0.00 | 0.05 ± 0.00 | 0.05 ± 0.00 |
L. interrogans serovar Canicola | 1.56 ± 0.00 | 12.50 ± 0.00 | 0.10 ± 0.00 | 0.20 ± 0.00 | 0.20 ± 0.00 | 0.39 ± 0.00 |
L. interrogans serovar Javanica | 3.13 ± 0.00 | 25.00 ± 0.00 | 0.78 ± 0.00 | 3.13 ± 0.00 | 0.78 ± 0.00 | 0.78 ± 0.00 |
Leptospira Serovars | Combination of AE and Doxycycline | Combination of ME and Doxycycline | ||||||
---|---|---|---|---|---|---|---|---|
MIC AE (µg/mL) | MIC DOX (µg/mL) | FICI | Activity | MIC ME (µg/mL) | MIC DOX (µg/mL) | FICI | Activity | |
L. interrogans serovar Australis | 800 ± 0.00 | 0.78 ± 0.00 | 1.59 ± 0.00 | Indifferent | 800 ± 0.00 | 0.39 ± 0.00 | 2.82 ± 0.00 | Indifferent |
L. interrogans serovar Bataviae | 50 ± 0.00 | 0.78 ± 0.00 | 3.09 ± 0.00 | Indifferent | 200 ± 0.00 | 0.39 ± 0.00 | 4.22 ± 0.00 | Antagonistic |
L. interrogans serovar Canicola | 100 ± 0.00 | 1.56 ± 0.00 | 2.09 ± 0.00 | Indifferent | 400 ± 0.00 | 1.56 ± 0.00 | 2.65 ± 0.00 | Indifferent |
L. interrogans serovar Javanica | 100 ± 0.00 | 1.56 ± 0.00 | 2.46 ± 0.00 | Indifferent | 400 ± 0.00 | 0.78 ± 0.00 | 3.58 ± 0.00 | Indifferent |
Leptospira Serovars | Combination of AE and Penicillin G | Combination of ME and Penicillin G | ||||||
---|---|---|---|---|---|---|---|---|
MIC AE (µg/mL) | MIC PEN (µg/mL) | FICI | Activity | MIC ME (µg/mL) | MIC PEN (µg/mL) | FICI | Activity | |
L. interrogans serovar Australis | 200 ± 0.00 | 0.02 ± 0.00 | 2.92 ± 0.00 | Indifferent | 800 ± 0.00 | 0.01 ± 0.00 | 3.32 ± 0.00 | Indifferent |
L. interrogans serovar Bataviae | 50 ± 0.00 | 0.02 ± 0.00 | 3.41 ± 0.00 | Indifferent | 400 ± 0.00 | 0.01 ± 0.00 | 2.95 ± 0.00 | Indifferent |
L. interrogans serovar Canicola | 100 ± 0.00 | 0.05 ± 0.00 | 2.42 ± 0.00 | Indifferent | 400 ± 0.00 | 0.05 ± 0.00 | 4.87 ± 0.00 | Antagonistic |
L. interrogans serovar Javanica | 100 ± 0.00 | 0.78 ± 0.00 | 2.61 ± 0.00 | Indifferent | 400 ± 0.00 | 0.78 ± 0.00 | 3.38 ± 0.00 | Indifferent |
Leptospira Serovars | Combination of AE and Ceftriaxone | Combination of ME and Ceftriaxone | ||||||
---|---|---|---|---|---|---|---|---|
MIC AE (µg/mL) | MIC CEF (µg/mL) | FICI | Activity | MIC ME (µg/mL) | MIC CEF (µg/mL) | FICI | Activity | |
L. interrogans serovar Australis | 400 ± 0.00 | 0.10 ± 0.00 | 2.39 ± 0.00 | Indifferent | 400 ± 0.00 | 0.20 ± 0.00 | 3.19 ± 0.00 | Indifferent |
L. interrogans serovar Bataviae | 100 ± 0.00 | 0.05 ± 0.00 | 2.24 ± 0.00 | Indifferent | 400 ± 0.00 | 0.02 ± 0.00 | 2.51 ± 0.00 | Indifferent |
L. interrogans serovar Canicola | 100 ± 0.00 | 0.20 ± 0.00 | 2.36 ± 0.00 | Indifferent | 100 ± 0.00 | 0.10 ± 0.00 | 3.22 ± 0.00 | Indifferent |
L. interrogans serovar Javanica | 200 ± 0.00 | 1.56 ± 0.00 | 2.40 ± 0.00 | Indifferent | 800 ± 0.00 | 1.56 ± 0.00 | 2.75 ± 0.00 | Indifferent |
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Ismail, C.A.M.; Deris, Z.Z.; Bakar, R.A.; Ismail, N. In Vitro Anti-Leptospiral Activity of Phyllanthus amarus Extracts and Their Combinations with Antibiotics. Int. J. Environ. Res. Public Health 2021, 18, 2834. https://doi.org/10.3390/ijerph18062834
Ismail CAM, Deris ZZ, Bakar RA, Ismail N. In Vitro Anti-Leptospiral Activity of Phyllanthus amarus Extracts and Their Combinations with Antibiotics. International Journal of Environmental Research and Public Health. 2021; 18(6):2834. https://doi.org/10.3390/ijerph18062834
Chicago/Turabian StyleIsmail, Che Ain Munirah, Zakuan Zainy Deris, Ruzilawati Abu Bakar, and Nabilah Ismail. 2021. "In Vitro Anti-Leptospiral Activity of Phyllanthus amarus Extracts and Their Combinations with Antibiotics" International Journal of Environmental Research and Public Health 18, no. 6: 2834. https://doi.org/10.3390/ijerph18062834