Assessment of the Phytochemicals and Antibacterial Potential of Azadirachta indica (Neem) Leaves Extract against Selected Clinical Isolates
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112-131
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Jun 6, 2026
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10.58578/ajstm.v3i2.10476
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Agbo Anthony Ogbonnia1, Mbah-Omeje Kelechi Nkechinyere2, Eze Vincent C.3 
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Abstract
The rapid emergence of antimicrobial resistance has intensified the search for alternative therapeutic agents from medicinal plants. Azadirachta indica (neem) is widely recognized for its broad-spectrum biological activities; however, comparative evidence on aqueous and ethanolic leaf extracts against clinical isolates, accompanied by phytochemical profiling, remains limited. This study aimed to evaluate the phytochemical constituents and antibacterial potential of aqueous and ethanolic leaf extracts of A. indica against selected clinical bacteria. Mature neem leaves were collected, authenticated, air-dried, pulverized, and extracted using distilled water and ethanol. The extracts were concentrated using a rotary evaporator. Qualitative and quantitative phytochemical analyses were conducted using standard procedures, while antibacterial activity was assessed through the agar well diffusion method against Escherichia coli, Shigella sp., Lactobacillus sp., and Staphylococcus aureus. Zones of inhibition were measured, and minimum inhibitory concentrations were determined. Phytochemical screening revealed the presence of alkaloids, saponins, flavonoids, tannins, and glycosides, with glycosides being the most abundant constituent (22.85 ± 0.084) and flavonoids the least abundant (6.55 ± 0.087). At 200 mg/mL, the aqueous extract demonstrated higher antibacterial activity than the ethanolic extract, with inhibition zones of 27 mm against E. coli, 26 mm against Shigella sp., 23 mm against Lactobacillus sp., and 20 mm against S. aureus. The minimum inhibitory concentration results indicated resistance of E. coli to both extracts and reduced sensitivity of Lactobacillus sp. to the ethanolic extract. These findings suggest that aqueous extraction may be more effective for harnessing the antibacterial properties of neem leaves. This study contributes to phytomedicine and antimicrobial research by highlighting the potential of A. indica leaf extract as a cost-effective plant-based antimicrobial agent for managing bacterial infections, particularly in resource-limited settings.
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Azadirachta indica; Antibacterial Activity; Phytochemical Constituents; Clinical Isolates; Antimicrobial Resistance
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Ogbonnia, A. A., Nkechinyere, M.-O. K., & C., E. V. (2026). Assessment of the Phytochemicals and Antibacterial Potential of Azadirachta indica (Neem) Leaves Extract against Selected Clinical Isolates. African Journal of Sciences and Traditional Medicine, 3(2), 112-131. https://doi.org/10.58578/ajstm.v3i2.10476
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
References
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Mickymaray, S. (2019). Efficacy and mechanism of traditional medicinal plants and bioactive compounds against clinically important pathogens. Antibiotics, 8(4), Article 257. https://doi.org/10.3390/antibiotics8040257
Muhammad, S. A., & Abubakar, S. M. (2016). Qualitative and quantitative determination of phytochemicals in aqueous extract of Chrysophyllum albidum seed kernel. Biosciences Biotechnology Research Asia, 13(2), 1201–1206. https://doi.org/10.13005/bbra/2153
Obodo, N. L., Tasie, F. O., & Mbah-Omeje, K. N. (2025). Antimicrobial activities of some medicinal plants used in folklore remedies in Enugu State, Nigeria. GSC Biological and Pharmaceutical Sciences. https://doi.org/10.30574/gscbps.2025.30.3.0020
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Tsaku, P. A., Ehinmidu, J. O., & Mohammed, S. A. (2017). Antibiotic susceptibility and plasmid profile of Escherichia coli from door handles in two tertiary institutions in Nasarawa State, Nigeria. Journal of Advances in Microbiology, 3(3), 1–13. https://doi.org/10.9734/JAMB/2017/33612
Vaou, N., Stavropoulou, E., Voidarou, C., Tsigalou, C., & Bezirtzoglou, E. (2021). Towards advances in medicinal plant antimicrobial activity: A review study on challenges and future perspectives. Microorganisms, 9(10), Article 2041. https://doi.org/10.3390/microorganisms9102041
Virshette, S. J., Patil, M. K., Deshmukh, A. A., Shaikh, J. R., & Dhas, M. S. (2020). Phytochemical constituents of different extracts of Azadirachta indica leaves in urine solvent of a non-pregnant cow. Journal of Pharmacognosy and Phytochemistry, 9(2), 1324–1328.
Waziri, M., & Saleh, I. A. (2015). Proximate analysis and phytochemical screening of Psidium guajava (guava) and Cucumis sativus (cucumber) grown in Gashua Fadama Area of Yobe State, Nigeria. International Research Journal of Pure and Applied Chemistry, 6(2), 77–83. https://doi.org/10.9734/IRJPAC/2015/13775
WHO. (2023). Traditional medicine has a long history of contributing to conventional medicine and continues to hold promise. https://www.who.int/news-room/feature-stories/detail/traditional-medicine-has-a-long-history-of-contributing-to-conventional-medicine-and-continues-to-hold-promise
Wifek, M., Saeed, A., Rehman, R., & Nisar, S. (2016). Lemon grass: A review on its botany, properties, applications and active components. International Journal of Chemical and Biochemical Sciences, 9, 79–84.
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Yildirim, A. B., Karakas, F. P., & Turker, A. U. (2013). In vitro antibacterial and antitumor activities of some medicinal plant extracts growing in Turkey. Asian Pacific Journal of Tropical Medicine, 6(8), 616–624. https://doi.org/10.1016/S1995-7645(13)60106-6
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Adewuyi, G. M., Iyevhobu, B. T., Momoh, A. M., & Samuel, S. O. (2024). Evaluation of African neem plant (Dogonyaro) antibacterial activity against certain pathogenic organisms. World Journal of Advanced Research and Reviews, 24(1), 2208–2214.
Agbo, A. O., Mbah-Omeje, K. N., Ikegwu, T. M., & Ameh, O. A. (2025). Proximate, phytochemical and gas chromatography flame ionization detection (GC-FID) profiling of ethanolic extract of guava (Psidium guajava) leaves and its antibacterial activities on some bacterial isolates. International Journal of Research and Innovation in Applied Sciences, 10(8), 1254–1298. https://doi.org/10.51584/IJRIAS.2025.100800111
Alekshun, M. N., & Levy, S. B. (2007). Molecular mechanisms of antibacterial multidrug resistance. Cell, 128(6), 1037–1050. https://doi.org/10.1016/j.cell.2007.03.004
Altemimi, A., Watson, D. G., Choudhary, R., Dasari, M. R., & Lightfoot, D. A. (2016). Ultrasound assisted extraction of phenolic compounds from peaches and pumpkins. PLoS ONE, 11(2), Article e0148758. https://doi.org/10.1371/journal.pone.0148758
Amadi, B., Emelieze, M., Agomuo, E., Ogunka-Nnoka, C., & Amadi, P. (2017). Proximate, GC-FID, and micronutrient analysis of extracts of Azadirachta indica. International Journal of Advanced Chemistry, 5(2), 73–79. https://doi.org/10.14419/ijac.v5i2.8124
Ankita, S., & Sapan, P. (2018). Preliminary phytochemical screening and quantitative analysis of secondary metabolites of Mentha arvensis and Azadirachta indica. International Journal of Advanced Research and Development, 3(1), 114–118.
Atanasov, A. G., Zotchev, S. B., Dirsch, V. M., & Supuran, C. T. (2021). Natural products in drug discovery: Advances and opportunities. Nature Reviews Drug Discovery, 20(3), 200–216. https://doi.org/10.1038/s41573-020-00114-z
Bamishaiye, E. I., Olayemi, F. F., Awagu, E. F., & Bamishaiye, O. M. (2011). Proximate and phytochemical composition of Moringa oleifera leaves at three stages of maturation. Advance Journal of Food Science and Technology, 3(4), 233–237.
Banerjee, S., Kim, L. M., Shariff, M., Khatoon, H., & Yusoff, F. M. (2013). Antibacterial activity of neem (Azadirachta indica) leaves on Vibrio spp. isolated from cultured shrimp. Asian Journal of Animal and Veterinary Advances, 8(2), 355–361. https://doi.org/10.3923/ajava.2013.355.361
Basnayake, P. I., & Gunatilake, M. (2024). Pharmacological effects of Pterocarpus marsupium. BLDE University Journal of Health Sciences, 9(1), 3–15. https://doi.org/10.4103/bjhs.bjhs_43_23
Clinical and Laboratory Standards Institute. (2012). Performance standards for antimicrobial susceptibility testing: Twenty-second informational supplement (CLSI document M100-S22). Clinical and Laboratory Standards Institute.
Cowan, M. M. (1999). Plant products as antimicrobial agents. Clinical Microbiology Reviews, 12(4), 564–582. https://doi.org/10.1128/cmr.12.4.564
Dhanani, T., Shah, S., Gajbhiye, N. A., & Kumar, S. (2017). Effect of extraction methods on yield, phytochemical constituents and antioxidant activity of Withania somnifera. Arabian Journal of Chemistry, 10, S1193–S1199. https://doi.org/10.1016/j.arabjc.2013.02.015
Gonelimali, F. D., Lin, J., Miao, W., Xuan, J., Charles, F., Chen, M., & Hatab, S. R. (2018). Antibacterial properties and mechanism of action of some plant extracts against food pathogens and spoilage microorganisms. Frontiers in Microbiology, 9, Article 1639. https://doi.org/10.3389/fmicb.2018.01639
Harborne, J. B. (1973). Phenolic compounds. In Phytochemical Methods (pp. 33–88). Springer. https://doi.org/10.1007/978-94-009-5921-7_2
Herrera-Calderon, O., Arroyo-Acevedo, J. L., Rojas-Armas, J., Chumpitaz-Cerrate, V., Figueroa-Salvador, L., Enciso-Roca, E., & Tinco-Jayo, J. (2017). Phytochemical screening, total phenolic content, antioxidant and cytotoxic activity of Chromolaena laevigata on human tumor cell lines. Annual Research & Review in Biology, 21(1), 1–9.
Kebede, T., Gadisa, E., & Tufa, A. (2021). Antimicrobial activities evaluation and phytochemical screening of some selected medicinal plants: A possible alternative in the treatment of multidrug-resistant microbes. PLoS ONE, 16(3), Article e0249253. https://doi.org/10.1371/journal.pone.0249253
Khanal, S. (2021). Qualitative and quantitative phytochemical screening of Azadirachta indica Juss. plant parts. International Journal of Applied Sciences and Biotechnology, 9(2), 122–127. https://doi.org/10.3126/ijasbt.v9i2.38050
Kumar, M., Tomar, M., Amarowicz, R., Saurabh, V., Nair, M. S., Maheshwari, C., Sasi, M., Prajapati, U., Hasan, M., Singh, S., Changan, S., Kumar, R., Prajapat, R. K., Berwal, M. K., & Satankar, V. (2021). Guava (Psidium guajava L.) leaves: Nutritional composition, phytochemical profile, and health-promoting bioactivities. Foods, 10(4), Article 752. https://doi.org/10.3390/foods10040752
Kunle, O. F., & Egharevba, H. O. (2013). Chemical constituents and biological activity of medicinal plants used for the management of sickle cell disease: A review. Journal of Medicinal Plants Research, 7(48), 3452–3476. https://doi.org/10.5897/JMPR2013.5333
Mamman, P. H., Mshelia, W. P., Susbatrus, S. C., & Sambo, K. W. (2013). Antibacterial effects of crude extract of Azadirachta indica against Escherichia coli, Salmonella spp., and Staphylococcus aureus. International Journal of Medicine and Medical Sciences, 5(1), 14–18. https://doi.org/10.5897/IJMMS12.017
Mehta, A., & Diwakar, M. (2021). Characterization and antimicrobial susceptibility profile of non-lactose fermenting gram-negative bacterial isolates in a tertiary care teaching hospital of central India. Asian Journal of Pharmaceutical and Clinical Research, 14(10), 1–7. https://doi.org/10.22159/ajpcr.2021.v14i10.42822
Mehta, A., Saxena, G., & Mani, A. (2016). Comparative analysis of antibacterial activity of aqueous, ethanolic, methanolic and acetone extracts of commercial green tea and black tea against standard bacterial strains. International Journal of Current Microbiology and Applied Sciences, 5(11), 145–152. https://doi.org/10.20546/ijcmas.2016.511.017
Mickymaray, S. (2019). Efficacy and mechanism of traditional medicinal plants and bioactive compounds against clinically important pathogens. Antibiotics, 8(4), Article 257. https://doi.org/10.3390/antibiotics8040257
Muhammad, S. A., & Abubakar, S. M. (2016). Qualitative and quantitative determination of phytochemicals in aqueous extract of Chrysophyllum albidum seed kernel. Biosciences Biotechnology Research Asia, 13(2), 1201–1206. https://doi.org/10.13005/bbra/2153
Obodo, N. L., Tasie, F. O., & Mbah-Omeje, K. N. (2025). Antimicrobial activities of some medicinal plants used in folklore remedies in Enugu State, Nigeria. GSC Biological and Pharmaceutical Sciences. https://doi.org/10.30574/gscbps.2025.30.3.0020
Prabagar, S., Nandakumar, Thuraisingam, S., & Prabagar, J. (2020). Antimicrobial activity of leaf and bark extracts of Cassia fistula. World Journal of Agricultural Sciences, 16(4), 227–232. https://doi.org/10.5829/idosi.wjas.2020.227.232
Radji, M., Agustama, R. A., Elya, B., & Tjampakasari, C. R. (2013). Antimicrobial activity of green tea extract against isolates of methicillin-resistant Staphylococcus aureus and multidrug-resistant Pseudomonas aeruginosa. Asian Pacific Journal of Tropical Biomedicine, 3(8), 663–667. https://doi.org/10.1016/S2221-1691(13)60133-1
Santosh, V. K., & Varadhan, V. (2013). Neem: Prehistory to contemporary medicinal uses to humankind. Asian Pacific Journal of Tropical Biomedicine, 3(7), 505–514. https://doi.org/10.1016/S2221-1691(13)60105-7
Shirajum, M., Luthfun, N., Manirul, I., Sharmin, S., Sarwar, H., Habiba, K., Humayun, K., Rubel, A., & Mammunar, R. (2015). Analgesic, anti-inflammatory and CNS activities of the methanolic extract of Artocarpus heterophyllus seed. World Journal of Pharmacy and Pharmaceutical Sciences, 4(5), 91–103.
Shrinet, K., Singh, K., Chaurasia, R. K., Tripathi, A. K., & Kumar, A. (2021). Bioactive compounds and their future therapeutic applications. In Natural Bioactive Compounds: Technological Advancements (pp. 337–355). Elsevier. https://doi.org/10.1016/B978-0-12-820655-3.00017-3
Stefanello, M. É. A., Cervi, A. C., Ito, I. Y., Salvador, M. J., Wisniewski, A., Jr., & Simionatto, E. L. (2008). Chemical composition and antimicrobial activity of essential oils of Eugenia chlorophylla (Myrtaceae). Journal of Essential Oil Research, 20(1), 75–78. https://doi.org/10.1080/10412905.2008.9699427
Tajkarimi, M. M., Ibrahim, S. A., & Cliver, D. O. (2010). Antimicrobial herb and spices compounds in food. Food Control, 21(9), 1199–1218. https://doi.org/10.1016/j.foodcont.2010.02.003
Tsaku, P. A., Ehinmidu, J. O., & Mohammed, S. A. (2017). Antibiotic susceptibility and plasmid profile of Escherichia coli from door handles in two tertiary institutions in Nasarawa State, Nigeria. Journal of Advances in Microbiology, 3(3), 1–13. https://doi.org/10.9734/JAMB/2017/33612
Vaou, N., Stavropoulou, E., Voidarou, C., Tsigalou, C., & Bezirtzoglou, E. (2021). Towards advances in medicinal plant antimicrobial activity: A review study on challenges and future perspectives. Microorganisms, 9(10), Article 2041. https://doi.org/10.3390/microorganisms9102041
Virshette, S. J., Patil, M. K., Deshmukh, A. A., Shaikh, J. R., & Dhas, M. S. (2020). Phytochemical constituents of different extracts of Azadirachta indica leaves in urine solvent of a non-pregnant cow. Journal of Pharmacognosy and Phytochemistry, 9(2), 1324–1328.
Waziri, M., & Saleh, I. A. (2015). Proximate analysis and phytochemical screening of Psidium guajava (guava) and Cucumis sativus (cucumber) grown in Gashua Fadama Area of Yobe State, Nigeria. International Research Journal of Pure and Applied Chemistry, 6(2), 77–83. https://doi.org/10.9734/IRJPAC/2015/13775
WHO. (2023). Traditional medicine has a long history of contributing to conventional medicine and continues to hold promise. https://www.who.int/news-room/feature-stories/detail/traditional-medicine-has-a-long-history-of-contributing-to-conventional-medicine-and-continues-to-hold-promise
Wifek, M., Saeed, A., Rehman, R., & Nisar, S. (2016). Lemon grass: A review on its botany, properties, applications and active components. International Journal of Chemical and Biochemical Sciences, 9, 79–84.
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