In vitro study of the antibacterial activity of ethanolic extracts of Punica granatum and Syzygium aromaticum
Keywords:
antibacterial, clove, gram-negative bacteria, gram-positive bacteria, plant extracts, pomegranateAbstract
Introduction: Antibiotic resistance in pathogenic bacteria is a growing phenomenon in both hospital and community settings, hindering medical treatment and clinical recovery of patients. An alternative to this problem lies in plant species that possess biological properties to combat microbial infections.
Objective: To determine the in vitro antibacterial activity of ethanolic extracts of Punica granatum L. and Syzygium aromaticum L.
Methods: An experimental investigation was conducted using ethanolic extracts of pomegranate and clove at concentrations of 100, 200, 300, 400, and 500 mg/mL, tested against strains of Staphylococcus aureus, Pseudomonas aeruginosa, and Vibrio cholerae; three replicates were performed for each assay. In vitro susceptibility was determined using the Kirby-Bauer disk diffusion and Müller-Hinton broth macrodilution techniques.
Results: All bacterial strains were sensitive to the pomegranate and clove extracts; the greatest antibacterial activity was observed against Staphylococcus aureus. At a concentration of 500 mg/mL, inhibitory zones of up to 20.5 and 20.0 mm in diameter were obtained. A minimum inhibitory concentration of 50 mg/mL was established for Staphylococcus aureus and 100 mg/mL for the other tested bacteria.
Conclusion: Ethanolic extracts made from Punica granatum L. peel and Syzygium aromaticum L. flowers have antibacterial activity related to the dose applied (greater than 500 mg/mL) against Staphylococcus aureus, Pseudomonas aeruginosa, and Vibrio cholerae.
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References
1. Camacho LA. Resistencia bacteriana, una crisis actual [Internet]. Rev Esp Salud Publica. 2023 [acceso: 08/06/2025]; 97: 1-10. Disponible en: https://pmc.ncbi.nlm.nih.gov/articles/PMC10541255/
2. Barrantes Jiménez K, Chacón Jiménez L, Arias AM. El impacto de la resistencia a los antibióticos en el desarrollo sostenible [Internet]. PSM. 2022 [acceso: 08/06/2025]; 19(2): 305-29. DOI: http://dx.doi.org/10.15517/psm.v0i19.47590
3. Organización Panamericana de la Salud. Patógenos multirresistentes que son prioritarios para la OMS - OPS/OMS [Internet]. OPS/OMS; 2021. [acceso: 08/06/2025]. Disponible en: https://www.paho.org/es/noticias/4-3-2021-patogenos-multirresistentes-que-son-prioritarios-para-om
4. Giono-Cerezo S, Santos-Preciado JI, Rayo Morfín-Otero Mdel, Torres-López FJ, Alcántar-Curiel MD. Resistencia antimicrobiana. Importancia y esfuerzos por contenerla [Internet]. Gac. Méd. Méx 2020;156(2): 172-80. DOI: https://doi.org/10.24875/gmm.20005624
5. Barros Gabriel MR, Melo E, Oliveira MI, Germinio JE, Santos Rosiel J, Oliveira Sibele R. Impacto financiero de la terapia con antibióticos en la resistencia a múltiples fármacos bacterianos en un hospital de emergencia en Pernambuco, Brasil [Internet]. Ars Pharm. 2020; 61(2): 121-6. DOI: https://dx.doi.org/10.30827/ars.v61i2.115337
6. Coopoosamy R, Singh K, Naidoo K, Nadasan DS. The role of phytomedicine: Bridging the gap between the past, present, and future [Internet]. Journal of Medicinal Plants for Economic Development. 2023; 7(1): 197. DOI: https://doi.org/10.4102/jomped.v7i1.197
7. Santa Cruz - López CY, Chapoñan M, Limo JA, Moreno MC. Susceptibilidad in vitro de bacterias patógenas a los extractos de Rosmarinus officinalis y Caesalpinia spinosa [Internet]. Rev Cubana Med Milit. 2023 [acceso: 12/06/2025];52(3): e02302933. Disponible en: https://revmedmilitar.sld.cu/index.php/mil/article/view/2933
8. Sarmiento Zurita AJ. Revisión: alternativas terapéuticas a los fármacos antimicrobianos por la problemática de multidrogoresistencia actual [Internet]. Rev Cient Cienc Méd. 2023 [acceso: 15/06/2025]; 26(2): 78-83. Disponible en: http://www.scielo.org.bo/scielo.php?pid=S1817-74332023000200078&script=sci_arttext
9. Pérez Aguedo D, Fornaris Pérez YA, Pérez AD. La medicina natural y tradicional como complemento de la terapia antimicrobiana [Internet]. Rev Cuban Med Mil. 2024 [acceso: 12/06/2025]; 53(2): e024025492. Disponible en: http://scielo.sld.cu/scielo.php?pid=S0138-65572024000200039&script=sci_arttext&tlng=pt
10. Mahmood MS, Ashraf A, Ali S, Siddique AB, Asad F, Abbas RZ, et al. Portrayal of Punica granatum L. peel extract through High Performance Liquid Chromatography and antimicrobial activity evaluation [Internet]. Braz J Biol. 2021 [acceso: 12/06/2025]; 83: e244435. Disponible en: http://www.scielo.br/j/bjb/a/TzBBtWkhvRZkK57JnpNRDbm/?lang=en
11. Bustamante A, García-Díaz D, Jiménez P, Valenzuela Rodrigo PME, Echeverría F. Potencial efecto terapéutico de los polifenoles obtenidos de la cáscara de granada en la esteatosis hepática [Internet]. Rev. chil. nutr. 2022; 49(1): 89-99. DOI: http://dx.doi.org/10.4067/S0717-75182022000100089
12. Batiha GES, Alkazmi LM, Wasef LG, Beshbishy AM, Nadwa EH, Rashwan EK. Syzygium aromaticum L. (Myrtaceae): Traditional Uses, Bioactive Chemical Constituents, Pharmacological and Toxicological Activities. Biomolecules [Internet]. 2020 [acceso: 12/06/2025];10(2): E202. Disponible en: https://pubmed.ncbi.nlm.nih.gov/32019140/
13. Haro-González JN, Castillo-Herrera GA, Martínez-Velázquez M, Espinosa-Andrews H. Clove essential oil (Syzygium aromaticum L. Myrtaceae): extraction, chemical composition, food applications, and essential bioactivity for human health [Internet]. Molecules. 2021 [acceso: 15/06/2025]; 26(21):6387. Disponible en: https://www.mdpi.com/1420-3049/26/21/6387
14. Ulanowska M, Olas B. Propiedades biológicas y perspectivas de aplicación del eugenol: una revisión [Internet]. Revista Internacional de Ciencias Moleculares. 2021; 22(7): 3671. DOI: https://doi.org/10.3390/ijms22073671
15. Kowalska-Krochmal B, Dudek-Wicher R. The minimum inhibitory concentration of antibiotics: methods, interpretation, clinical relevance [Internet]. Pathogens. 2021; 10(2):165. DOI: https://doi.org/10.3390/pathogens10020165
16. Sacsaquispe R. Manual de procedimientos para la prueba de sensibilidad antimicrobiana por el método de Disco Difusión [Internet]. Perú: Instituto Nacional de Salud; 2002. [acceso: 15/06/2025]. Disponible en: https://cdn.www.gob.pe/uploads/document/file/417394/439893732843877347520191106-32001-1v6txak.pdf?v=1573077224
17. Thakur M, Khushboo YA, Dubey KK, Dakal TC, Yadav V. Antimicrobial activity against antibiotic-resistant pathogens and antioxidant activity and LCMS/MS phytochemical content analysis of selected medicinal plants [Internet]. J Pure Appl Microbiol. 2024;18(1):722-38. DOI: https://doi.org/10.22207/JPAM.18.1.62
18. Serquén LM, Mantilla MCM, Osores SAI. Susceptibilidad de «Candida albicans» a extracto etanólico de cáscara de «Punica granatum» [Internet]. Med Natur. 2020 [acceso: 19/06/2025]; 14(1):59-64. Disponible en: https://dialnet.unirioja.es/servlet/articulo?codigo=7248977
19. Faujdar SS, Bisht D, Sharma A. Antibacterial activity of Syzygium aromaticum (clove) against uropathogens producing ESBL, MBL, and AmpC beta-lactamase: Are we close to getting a new antibacterial agent? [Internet]. J Fam Med Prim Care. 2020 [acceso: 21/06/2025]; 9(1):180-6. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7014883/
20. Kupnik K, Primožič M, Vasić K, Knez Ž, Leitgeb M. A Comprehensive Study of the antibacterial activity of bioactive juice and extracts from pomegranate (Punica granatum L.) Peels and Seeds [Internet]. Plants. 2021 [acceso: 19/06/2025]; 10(8):1554. Disponible en: https://pubmed.ncbi.nlm.nih.gov/34451599/
21. Troncoso C, Pavez M, Santos A, Salazar R, Barrientos L, Troncoso C, et al. Implicancias estructurales y fisiológicas de la célula bacteriana en los mecanismos de resistencia antibiótica [Internet]. Int J Morphol. 2017 [acceso: 23/06/2025]; 35(4):1214-23. Disponible en: https://www.scielo.cl/scielo.php?pid=S0717-95022017000401214&script=sci_arttext
22. Ghai I. Una barrera de entrada: Análisis de la membrana externa bacteriana y la resistencia a los antibióticos [Internet]. Ciencias Aplicadas. 2023 [acceso: 23/06/2025]; 13(7): 4238. DOI: https://doi.org/10.3390/app13074238
23. Scaglione E, Sateriale D, Mantova G, Di Rosario M, Continisio L, Vitiello M, et al. Antimicrobial efficacy of Punica granatum Lythraceae peel extract against pathogens belonging to the ESKAPE group [Internet]. Front Microbiol. 2024;15: 1383027. DOI: https://doi.org/10.3389/fmicb.2024.1383027
24. Maggini V, Semenzato G, Gallo E, Nunziata A, Fani R, Firenzuoli F. Antimicrobial activity of Syzygium aromaticum essential oil in human health treatment [Internet]. Molecules. 2024; 29(5): 999. DOI: https://doi.org/10.3390/molecules29050999
25. Noreen S, Hashmi B, Aja PM, Atoki AV. Phytochemicals and pharmacology of pomegranate (Punica granatum L.): nutraceutical benefits and industrial applications: a review [Internet]. Front Nutr. 2025;12: 1528897. DOI: https://doi.org/10.3389/fnut.2025.1528897
26. Vivot EP, Sánchez C, Cacik F, Sequin C. Actividad antibacteriana en plantas medicinales de la flora de Entre Ríos (Argentina) [Internet]. Cienc Docencia Tecnol. 2012 [acceso: 19/06/2025]; 45:131-46. Disponible en: https://www.scielo.org.ar/scielo.php?pid=S1851-17162012000200008&script=sci_arttext&tlng=pt
27. Martínez SM, Jiménez M del C, García PE, Figueroa M, Maceiras MA, Martínez SM, et al. Actividad cicatrizante in vivo del polvo carbonizado de Punica granatum Linn y Eichhornia crassipes [Internet]. Rev Cuba Med Mil. 2019 [acceso: 21/06/2025]; 48(2):166-76. Disponible en: http://scielo.sld.cu/pdf/mil/v48n2/1561-3046-mil-48-02-e284.pdf
28. Qian W, Sun Z, Wang T, Yang M, Liu M, Zhang J, et al. Antimicrobial activity of eugenol against carbapenem-resistant Klebsiella pneumoniae and its effect on biofilms [Internet]. Microb Pathog. 2020 [acceso: 21/06/2025]; 139: 103924. Disponible en: https://www.sciencedirect.com/science/article/abs/pii/S0882401019316948?via%3Dihu
29. Ginting EV, Retnaningrum E, Widiasih DA. Antibacterial activity of clove (Syzygium aromaticum) and cinnamon (Cinnamomum burmannii) essential oil against extended-spectrum β-lactamase-producing bacteria [Internet]. Vet World. 2021 [acceso: 21/06/2025];14(8):2206-11. Disponible en: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8448639/
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