Antimicrobial Activity of Crude Methanolic Extract Fractions from Balanites aegyptiaca (L.) Delile
Keywords:
Plant phenolics, Phenolic extracts, Antimicrobial activity, Phytochemicals, and Balanites aegyptiacaAbstract
Microbial infections remain a major global health concern, with mortality rates continuing to rise each decade. Projections estimate that by 2050, antimicrobial resistance could account for up to 10 million deaths annually. This study investigated the antimicrobial potential of phytochemical groups present in crude extracts of B. aegyptiaca as a possible alternative for managing microbial infections. Plant materials (leaves, stem bark, and root bark) were collected from two local varieties of B. aegyptica which are tentatively referred to as the “sweet” and “sour” varieties growing in Ajeluk village, Katakwi District, North-Eastern Uganda. The dried and pulverized samples (100 g each) were subjected to successive extraction with n-hexane, dichloromethane, and methanol. The resulting crude extracts were concentrated using a rotary evaporator, followed by fractionation through silica gel column chromatography. Fractions were further purified using preparative thin-layer chromatography, with separated bands visualized under Ultra Violet (UV) light, eluted, and tested for antimicrobial activity. Test organisms included multidrug-resistant American Type Culture Collection (ATCC) Escherichia coli ATCC 25922, Candida albicans ATCC 10231, and Aspergillus flavus ATCC 6275. The methanolic fractions demonstrated notable antimicrobial activity, with inhibition zones greater than 8 mm. Among the phytochemical groups, phenolics exhibited the strongest activity (23.67 ± 1.15 mm), while coumarin-containing fractions displayed the lowest activity (7.00 ± 0.00 mm). Statistical analysis revealed significant differences between inhibition zones of the phytochemical groups and the positive control (p < 0.05). Phytochemical screening confirmed the presence of phenolics, flavonoids, alkaloids, cardiac glycosides, and coumarins in the extracts. Overall, the findings suggest that B. aegyptica contains diverse bioactive compounds with antimicrobial potential, particularly against fungal pathogens. C. albicans and A. flavus were more susceptible to these extracts than E. coli, highlighting the plant’s promise as a source of antifungal agents.
References
Annunziata, F., Pinna, C., Dallavalle, S., Tamborini, L., & Pinto, A. (2020). An overview of coumarin as a versatile and readily accessible scaffold with broad-ranging biological activities. International Journal of Molecular Sciences, 21(14), 4618. https://doi.org/10.3390/ijms21134618
Bengtsson-Palme, J., Kristiansson, E., & Larsson, D. G. J. (2018). Environmental factors influencing the development and spread of antibiotic resistance. FEMS Microbiology Reviews, 42(1), 68–80. https://doi.org/10.1093/femsre/fux053
Bouarab-Chibane, L., Forquet, V., Lantéri, P., Clément, Y., Léonard-Akkari, L., Oulahal, N., Degraeve, P., & Bordes, C. (2019). Antibacterial properties of polyphenols: Characterization and QSAR (quantitative structure–activity relationship) models. Frontiers in Microbiology, 10, 829. https://doi.org/10.3389/fmicb.2019.00829
Cheke, R. S., Patel, H. M., Patil, V. M., Ansari, I. A., Shinde, S. D., Kadri, A., Snoussi, M., Adnan, M., Ambhore, J. P., Kharkar, P. S., Pasupuleti, V. R., & Deshmukh, P. K. (2022). Molecular insights into coumarin analogues as antimicrobial agents: Recent developments in drug discovery. Antibiotics, 11(5), 566. https://doi.org/10.3390/antibiotics11050566
Cushnie, T. P., Cushnie, B., & Lamb, A. J. (2014). Alkaloids: An overview of their antibacterial, antibiotic-enhancing and antivirulence activities. International Journal of Antimicrobial Agents, 44(5), 377–386. https://doi.org/10.1016/j.ijantimicag.2014.06.001
Eapen, M. S., Shukla, S. D., & Sohal, S. S. (2019). Flavonoids: A potential therapy for chronic diseases. Frontiers in Pharmacology, 10, 1206. https://doi.org/10.3389/fphar.2019.01206
Esposito, F., Carli, I., Del Vecchio, C., Xu, L., Corona, A., Grandi, N., Piano, D., Maccioni, E., Distinto, S., Parolin, C., & Tramontano, E. (2016). Sennoside A, derived from the traditional Chinese medicine plant Rheum L., is a new dual HIV-1 inhibitor effective on HIV-1 replication. Phytomedicine, 23(12), 1383–1391. https://doi.org/10.1016/j.phymed.2016.08.001
Gizaw, S. T., Tsegaye, A. T., & Asfaw, T. G. (2022). Cardiac glycosides: Natural products with potential antimicrobial applications. Journal of Ethnopharmacology, 282, 114626. https://doi.org/10.1016/j.jep.2021.114626
Ibraheem, O., & Maimako, R. F. (2014). Evaluation of alkaloids and cardiac glycosides contents of Ricinus communis Linn. (Castor) whole plant parts and determination of their biological properties. International Journal of Toxicological and Pharmacological Research, 6(3), 34–42.
Jia, C., Zhang, J., Yu, L., Wang, C., Yang, Y., Rong, X., Xu, K., & Chu, M. (2019). Antifungal activity of coumarin against Candida albicans is related to apoptosis. Frontiers in Cellular and Infection Microbiology, 8, 445. https://doi.org/10.3389/fcimb.2018.00445
Khameneh, B., Eskin, N. A. M., Iranshahy, M., & Fazly Bazzaz, B. S. (2021). Phytochemicals: A promising weapon in the arsenal against antibiotic-resistant bacteria. Antibiotics, 10(9), 1044. https://doi.org/10.3390/antibiotics10091044
Khamis, G., Saleh, A. M., Habeeb, T. H., Hozzein, W. N., Wadaan, M. A. M., Papenbrock, J., & Abdelgawad, H. (2020). Provenance effect on bioactive phytochemicals and nutritional and health benefits of the desert date Balanites aegyptiaca. Journal of Food Biochemistry, 44(1), e13229. https://doi.org/10.1111/jfbc.13229
Lee, J.-H., Kim, Y.-G., Park, I., & Lee, J. (2024). Antifungal and antibiofilm activities of flavonoids against Candida albicans: Focus on 3,2′-dihydroxyflavone as a potential therapeutic agent. Biofilm, 8, 100218. https://doi.org/10.1016/j.bioflm.2024.100218
Lobiuc, A., Pavăl, N.-E., Mangalagiu, I. I., Gheorghiță, R., Teliban, G.-C., Amriucai-Mantu, D., & Stoleru, V. (2023). Future antimicrobials: Natural and functionalized phenolics. Molecules, 28(3), 1114. https://doi.org/10.3390/molecules28031114
López-Jacome, L. E., Mercado-Casillas, Y. M., Méndez-Sotelo, B. J., Jiménez-Cortes, J. G., Tovar-García, A., Estrada-Velasco, A. Y., Almeida-Villegas, J. A., Martínez, J. D. P., & García-Contreras, R. (2022). Anti-bacterial agents. In Encyclopedia of Infection and Immunity (Vol. 4, pp. 494–509). https://doi.org/10.1016/B978-0-12-818731-9.00186-5
Mudenda, S., Chabalenge, B., Daka, V., Mfune, R. L., Salachi, K. I., Mohamed, S., Mufwambi, W., & Kasanga, M. (2023). Global strategies to combat antimicrobial resistance: A One Health perspective. Psychology and Psychiatry, 14(8), 271–328. https://doi.org/10.4236/pp.2023.148020
Namubiru, S., Migisha, R., Okello, P. E., Simbwa, B., Kabami, Z., Agaba, B., Zalwango, J. F., Naiga, H. N., Zalwango, M. G., Wanyana, M. W., Monje, F., King, P., Kawungezi, P. C., Kiggundu, T., Ninsiima, M., Akunzirwe, R., Namusosa, R., Mugerwa, I., Winfred, A. D., & Nabadda, S. (2024). Increasing trends of antibiotic resistance in Uganda: Analysis of the national antimicrobial resistance surveillance data, 2018–2021. BMC Infectious Diseases, 24(1), 1–17. https://doi.org/10.1186/s12879-024-09806-y
Othman, L., Sleiman, A., & Abdel-Massih, R. M. (2019). Antimicrobial activity of polyphenols and alkaloids in Middle Eastern plants. Frontiers in Microbiology, 10, 911. https://doi.org/10.3389/fmicb.2019.00911
Oulahal, N., & Degraeve, P. (2022). Phenolic-rich plant extracts with antimicrobial activity: An alternative to food preservatives and biocides? Frontiers in Microbiology, 12, 753518. https://doi.org/10.3389/fmicb.2021.753518
Parham, S., Kharazi, A. Z., Bakhsheshi-Rad, H. R., Nur, H., Ismail, A. F., Sharif, S., & Berto, F. (2020). Antioxidant, antimicrobial and antiviral properties of herbal materials. Antioxidants, 9(12), 1309. https://doi.org/10.3390/antiox9121309
Policegoudra, R. S., Saikia, S., Das, J., Chattopadhyay, P., Singh, L., & Veer, V. (2012). Phenolic content, antioxidant activity, antibacterial activity and phytochemical composition of Garcinia lancifolia. Indian Journal of Pharmaceutical Sciences, 74(3), 268–271.
Pudi, N., Varikuti, G. D., Badana, A. K., Gavara, M. M., Kumari, S., & Malla, R. (2016). Studies on optimization of growth parameters for enhanced production of antibiotic alkaloids by isolated marine actinomycetes. Journal of Applied Pharmaceutical Science, 6(10), 181–188. https://doi.org/10.7324/JAPS.2016.601025
Reen, F. J., Gutiérrez-Barranquero, J. A., Parages, M. L., & O’Gara, F. (2018). Coumarin: A novel player in microbial quorum sensing and biofilm formation inhibition. Applied Microbiology and Biotechnology, 102(5), 2063–2073. https://doi.org/10.1007/s00253-018-8787-x
Shamsudin, N. F., Ahmed, Q. U., Mahmood, S., Ali Shah, S. A., Khatib, A., Mukhtar, S., Alsharif, M. A., Parveen, H., & Zakaria, Z. A. (2022). Antibacterial effects of flavonoids and their structure-activity relationship study: A comparative interpretation. Molecules, 27(4), 1149. https://doi.org/10.3390/molecules27041149
Singh, P. K., Singh, J., Medhi, T., & Kumar, A. (2022). Phytochemical screening, quantification, FT-IR analysis, and in silico characterization of potential bio-active compounds identified in HR-LC/MS analysis of the polyherbal formulation from Northeast India. ACS Omega, 7(37), 33067–33078. https://doi.org/10.1021/acsomega.2c03117
Stan, D., Enciu, A.-M., Mateescu, A. L., Ion, A. C., Brezeanu, A. C., Stan, D., & Tanase, C. (2021). Natural compounds with antimicrobial and antiviral effect and nanocarriers used for their transportation. Frontiers in Pharmacology, 12, 723233. https://doi.org/10.3389/fphar.2021.723233
Škubník, J., Pavlíčková, V., & Rimpelová, S. (2021). Cardiac glycosides as immune system modulators. Biomolecules, 11(5), 659. https://doi.org/10.3390/biom11050659
Tagousop, C. N., Tamokou, J.-d.-D., Ekom, S. E., Ngnokam, D., & Voutquenne-Nazabadioko, L. (2018). Antimicrobial activities of flavonoid glycosides from Graptophyllum grandulosum and their mechanism of antibacterial action. BMC Complementary and Alternative Medicine, 18(1), 252. https://doi.org/10.1186/s12906-018-2321-7
Takó, M., Kerekes, E. B., Zambrano, C., Kotogán, A., Papp, T., Krisch, J., & Vágvölgyi, C. (2020). Plant phenolics and phenolic-enriched extracts as antimicrobial agents against food-contaminating microorganisms. Antioxidants, 9(2), 165. https://doi.org/10.3390/antiox9020165
Thornber, K., Verner-Jeffreys, D., Hinchliffe, S., Rahman, M. M., Bass, D., & Tyler, C. R. (2020). Evaluating antimicrobial resistance in the global shrimp industry. Reviews in Aquaculture, 12(2), 966–986. https://doi.org/10.1111/raq.12367
Thebti, A., Meddeb, A., Ben Salem, I., Bakary, C., Ayari, S., Rezgui, F., Essafi-Benkhadir, K., Boudabous, A., & Ouzari, H.-I. (2023). Antimicrobial activities and mode of flavonoid actions. Antibiotics, 12(2), 225. https://doi.org/10.3390/antibiotics12020225
Umar, A. (2023). Evaluation of antimicrobial activity and phytochemical analysis of ethanolic leaf extract of Balanites aegyptiaca. International Research Journal of Innovations in Engineering and Technology, 7(7), 178–181. https://doi.org/10.47001/IRJIET/2023.707027
Usman, A., Mohammed, Y., & Opaluwa, O. D. (2020). Phytochemicals, antibacterial, antioxidant and cytotoxic analyses of Balanites aegyptiaca stem bark extracts. Nigerian Research Journal of Chemical Sciences, 8(1).
Villanueva, X., Zhen, L., Ares, J. N., Vackier, T., Lange, H., Crestini, C., & Steenackers, H. P. (2023). Effect of chemical modifications of tannins on their antimicrobial and antibiofilm effect against Gram-negative and Gram-positive bacteria. Frontiers in Microbiology, 13, 987164. https://doi.org/10.3389/fmicb.2022.987164
Wu, T., He, M., Zang, X., Zhou, Y., Qiu, T., Pan, S., & Xu, X. (2013). A structure–activity relationship study of flavonoids as inhibitors of E. coli by membrane interaction effect. Biochimica et Biophysica Acta (BBA) – Biomembranes, 1828(11), 2751–2756. https://doi.org/10.1016/j.bbamem.2013.07.029
Yan, Y., Li, X., Zhang, C., Lv, L., Gao, B., & Li, M. (2021). Research progress on antibacterial activities and mechanisms of natural alkaloids: A review. Antibiotics, 10(3), 318. https://doi.org/10.3390/antibiotics10030318