• AbdulGaniy Agbaje Department of Biological Sciences, Al-Hikmah University, Ilorin
  • Maryam Moyosore Muhammed
  • Sharafa Adegoke Bello
  • Oluwabukola Kudirat Jimoh-Hamza
  • Muinat Olanike Kazeem
  • Zainab Oluwatoyin Azeez
  • Khadijat Ndagi Jibril
Keywords: Azadirachta indica, Methanolic, Synthesis, Silver nanoparticles, MIC, MBC


Azadirachta indica is a plant commonly known as neem. It is native to the Indian subcontinent and most countries in Africa, including Nigeria. The aim of this study was to synthesise silver nanoparticles and examine antimicrobial activities of crude methanolic extract of Azadirachta indica leaves on clinical isolates of Staphylococcus aureus, Escherichia coli and Candida albicans. Methanol was the solvent used for the extraction. The characterization of synthesized AgNPs was carried out using the UV-Visible spectrophotometer. Antimicrobial activity was conducted using disc diffusion method while the Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Minimum Fungicidal Concentration (MFC) were determined using broth dilution method. The antimicrobial activity of the synthesized Azadirachta indica AgNPs was carried out using standard agar well diffusion method. Qualitative phytochemicals analysis revealed presence of tannins, phenols, alkaloids, phytosteroid, and terpenoid while flavonoids, steroids and saponin were absent.  Antibacterial activities of the extracts using the ditch method (concentrations of 100mg/mL, 50mg/mL, 25mg/mL, 12.5mg/mL, and 6.25mg/mL) recorded no zone of inhibition for Staphylococcus aureus and Escherichia coli. Antimicrobial activity of synthesized silver nanoparticles (AgNPs) from methanolic extract showed no zone of inhibition for Staphylococcus aureus. The zone of inhibition against Escherichia coli was 5.33mm while for Candida albicans the zone of inhibition was 15.33mm. Based on the results obtained from this study, it could be said that methanolic extract of Azadirachta indica leaves contains phytochemicals of pharmacological significance which could be exploited as antifungal agent.


Ahmed, S., Ahmad, M., Swami, B.L., Ikram, S. (2015). Plants extract mediated synthesis of silver nanoparticles for antimicrobial applications: a green expertise. Journal of Advanced Research.02.007.

Akinjogunla, O.J., Ekoi, O.H., Odeyemi, A.T., Etok, C.A., Oshoma, CE. (2011). Preliminary phytochemistry and in-vitro antibacterial efficacy of Hydro-ethanolic leaf extracts of Psidiumguajava on common urinary tract bacterial pathogens. Biology research Bulletin;5:329-336.

Akiyamam, H., Fujii, K., Yamasaki, O., Oono, T., Iwatsuki, K. (2001). Antibacterial action of several tannins against staphylococcus aureus. Journal of antimicrobial chemotherapy.48(4):487-491. DOI:

Akter, R., Mahabub-Uz-Zaman, M., Rahman, M.S. (2013). “Comparative studies on antidiabetic effect with phytochemical screening of Azadirachta indica and Andrographis paniculata,” IOSR Journal of Pharmacy and Biological Sciences, vol. 5, no. 2,pp. 122–128. DOI:

Banso, A and Ayodele, O.P (2001). Activities of extracts Garcinakola against Escherichia coli and Aspergillus niger. Journal of applied science and management,5:58-65.

Biswas, K., Chattopadhyay, I., Banerjee, R.K., Bandyopadhyay, U. (2002). Biological activities and medicinal properties of Neem (Azadirachta indica). Current Science, 82:1336-1345.

Chernousova, S. and Epple, M. (2013). Silver as antibacterial agent: Ion, nanoparticle, and metal. Angewandte Chemie International Edition, 52, 1636–1653. DOI:

Cowman, M.M (1999). Plant products as antimicrobial agents. Clinical microbiology review, 12(4):546-582. DOI:

Das, A.I, Kumar, R., Goutam, S.P. and Sagar, S.S (2016). Sunlight irradiation induced synthesis of silver nanoparticles using Glycolipid Bio-surfactant and exploring the antibacterial activity. Journal of Biological engineering and Biomedical sciences. 6(208), 2-10. DOI:

Del-Rio, A., Obdululio, B.G., Casfillo, J., Main, F.G. Ortuno, A. (1997). Uses and properties of citrus flavonoids. Journal of agricultural food chemical, 45:4505-4515. DOI:

Desmarchelier, P. and Fegan, N. (2002). Encyclopedia of Dairy Sciences.

Foster, T. J. (2002). Staphylococcus aureus. Molecular Medical Microbiology, 839-888. DOI:

Ghonmode, W.N., Balsaraf, O.D., Tambe, V.H., Saujanya, K.P., Patil, A.K., and Kakde, D.D. (2013). “Comparison of the antibacterial efficiency of neem leaf extracts, grape seed extracts and 3% sodium hypochlorite against E. feacalis—an in vitro study,”Journal of International Oral Health, vol. 5, no. 6, pp. 61–66.

Gurunathan, S., Han, J.W., Kim, E.S., Park, J.H., Kim, J.H. (2015). Reduction of graphene oxide by resveratrol:A novel and simple biological method for the synthesis of an effective anticancer nanotherapeutic molecule. International Journal of Nanomedicine 10, 2951–2969. DOI:

Jassim, A.M.N., Mohammed, M.T., Farhan, S.A., Dadoosh, R.M., Majeed, Z.N, Abdula, A.M. (2019). Green synthesis of silver nanoparticles using Carica papaya juice and study of their biochemical application. Journal of Pharmaceutical Science Research. 11:1025-1034.

Kalainila, P., Subha, V., Ernest Raindran, R.S., Renganathan, S. (2014).Synthesis and characterization of silver nanoparticles from Erythrina indica. Asian Journal of pharmaceutical clinical research, 7(2).

Kareem, S.O., Akpan, I., Ojo, O.P. (2008). Antimicrobial activities of Calotropis procera on selected pathogenic microorganisms. Africa journal of Biomedical research.

Kim, K.J., Sung, W.S., Moon, S.K., Choi, J.S., Kim, J.G., Lee, D.G. (2008). Antifungal effect of silver nanoparticles on dermatophytes. Journal of Microbiology and Biotechnology.18, 1482–1484

Lai, R., Sankarnaryanan, A., Mathur, V.S., Sharma, P.L. (2005). Antifertility effect of Neem oil in female albino rats by Intra vaginal and oral routes. Indian Journal of Medicine, 83:89-92.

Lateef, A., Azeez, M.A., Asafa, T.B., Yekeen, T.A., Akinboro, A., Oladipo, I.C., Azeez, L., Ajibade, S.E., Ojo, S.A., Gueguim-Kana, E.B., Beukes, L.S. (2016a). Biogenic synthesis of silver nanoparticles using a pod extract of Cola nitida: Antibacterial, antioxidant activities and application as a paint additive. Journal of Taibah University of Science 10:551-562. DOI:

Lateef, A., Akande, M.A., Azeez, M.A., Ojo, S.A., Folarin, B.I., Gueguim-Kana, E.B., Beukes, L.S. (2016b). Phytosynthesis of silver nanoparticles (AgNPs) using miracle fruit plant (Synsepalum dulcificum) for antimicrobial, catalytic, anticoagulant and thrombolytic applications. Nanotechnol Rev. 5:507-520. DOI:

Lateef, A., Akande ,M.A., Ojo, S.A., Folarin, B.I., Gueguim-Kana, E.B., Beukes, L.S. (2016c). Paper wasp nest-mediated biosynthesis of silver nanoparticles for antimicrobial, catalytic, anticoagulant and thrombolytic applications. 3Biotech. 6: 140. DOI:

Li, C.Y., Zhang, Y.J., Wang, M., Zhang, Y., Chen, G., Li, L., Wu, D., Wang, Q. (2014). In vivo real-time visualization of tissue blood flow and angiogenesis using Ag2S quantum dots in the NIR-II window. Biomaterials 35,393–400. DOI:

Mahfuzul, M.D., Hoque, M.L., Bari, Y., Inatsu, V.K., Juneja, Kawamoto, S. (2007).“Antibacterial activity of guava (Psidium guajava L.) and neem (Azadirachta indica A. Juss.) extracts against foodborne pathogens and spoilage bacteria,” Foodborne Pathogens and Disease, vol. 4, no. 4, pp. 481–488. DOI:

Mody, V.V., Siwale, R., Singh, A., Mody, H.R. (2010) Introduction to metallic nanoparticles. Journal of Pharmacy and Bioallied Sciences 2:282–289. DOI:

Mondali, N. K., Mojumdar, A., Chatterje, S. K., Banerjee, A., Datta, J. K., Gupta, S. (2009). Antifungal activities and chemical characterization of Neem leaf extracts on the growth of some selected fungal species in vitro culture medium. Journal of Applied Sciences and Environmental Management. 13(1):49–53.

Ogar, A., Tylko, G., Turnau, K. (2015). Antifungal properties of silver nanoparticles against indoor mould growth. Science of the Total Environment 521, 305–314. DOI:

Ogbuewu, I.P., Odoemenam, V.U., Obikaonu. H.O., Opara, M.N., Emenalom, O.O., Uchegbu M.C. (2011). The Growing Importance of Neem (Azadirachta indica A. Juss) In Agriculture, Industry, Medicine and Envirnment: A Review, Research Journal of Medicinal Plant 5(3),230-245. DOI:

Okwu, D.E (2004). Phytochemical and vitamins content of indigenous species of South-eastern Nigeria. Journal of sustain agricultural environment . 6(1):30-37.

Parekh, J and Chinda, S. (2008). Phytochemical screening of some plants from Western region of Indian plans. 8:657-662.

Parotta J.A., (2001). Healing plants of Peninsular India., New York, CABI Publishing, 495-96. DOI:

Ross I.A., Medicinal plants of the world: Chemical constituents, Traditional and modern medicinal uses, Totowa, New Jersey, 2, 81-85.

Peralta-Videa, J.R., Huang, Y., Jason, G., Parsons, J.G., Lijuan Zha, L., Lopez-Moreno, L.L., Hernandez-Viezcas, J.A., GardeaTorresdey, J.L. (2016). Plant-based green synthesis of metallic nanoparticles: scientific curiosity or a realistic alternative to chemical synthesis? Nanotechnology Environmental Engineer. 1:332. DOI:

Qayyum, S., Khan, A.U. (2016). Biofabrication of broad range antibacterial and antibiofilm silver nanoparticles. IET Nanobiotechnology 10:349-357. DOI:

Riar, S.S., Devkumar, C., Ilavazhagam, G., Bardham, J. (2007). Volatile fraction of Neem oil as a spermicide. Contraception (42):479-487. DOI:

Saha, J., Begum, A., Mukherjee, A., Kumar, S. (2017). A novel green synthesis of silver nanoparticles and their catalytic action in reduction of Methylene Blue dye. Sustainable Environment Research 554-557. DOI:

Salah, N., Miller, N.J., Pagange, G., Tijburg, L., Bowell, G.P. (1995). Polyphenolic flavonoids as scarvenger of aqueous phase radicals as chai breaking antioxidant. Archives of Biochemical and Biophysics 2:339-346. DOI:

Saratale, R.G, Benelli, G., Kumar, G. (2018). Bio-fabrication of silver nanoparticles using the leaf extract of an ancient herbal medicine, dandelion (Taraxacum officinale), evaluation of their antioxidant, anticancer potential and antimicrobial activity against phytopathogens. Environment Science Pollution Research. 25:10392-10406. DOI:

Saseed, A., Khan, Junaid Aslam . (2008). Study on the effect of Neem (Azhadirachta indica) leaves smoke in controlling airborne Bacteria in Residential premises. Current research in Bacteriology1 (2): 64-66. DOI:

Sathiyavimal, S., Vasantharaj, S., Bharathi, D., Saravanan, M., Manikandan, E., Kumar, S.S., Pugazhendhi, A. (2018). Biogenesis of copper oxide nanoparticles (CuONPs) using Sida acuta and their incorporation over cotton fabrics to prevent the pathogenicity of gram negative and gram positive bacteria. Journal of Photochemical Photobiological. 188: 126–134. DOI:

Sexton, J.A., Brown, V. and Johnston, M. (2007). “Regulation of sugar transport and metabolism by the Candida albicans Rgt1transcriptional repressor,” Yeast , vol. 24, no. 10 pp.847-860. DOI:

Sharma, V.K., Yngard, R.A., Lin, Y. (2009). Silver nanoparticles: Green synthesis and their antimicrobial activities. Advances in Colloid and Interface, 145, 83–96. DOI:

Shravan, Kumar, Mankala, Kannappan, Nagappan. (2011). In vivo Antidiabetic evaluation of Neem leaf extract in alloxan induced rats. Journal of applied Pharmaceutical science, 7, 100-105.

Sonia, Bajaj and Srinivasan, B.P. (2009). Investigation into the Anti diabetic activity of Azadirachta indica. Indian journal of pharmacology 31:138-141.

Tucker, G. S. (2022). Food preservation and biodeterioration. John Wiley & Sons.

Varghese, S., Narmadha, R., Gomathi, D., Kalaiselvi, M., Devaki, K., (2013). Phytochemical screening and HPTLC finger printing analysis of Citrus luslanatus (Thumb) seed. Journal of acute disease 2.(2) 122-126. DOI:

Vasantharaj, S., Sripriya, N., Shanmugavel, M., Manikandan, E., Gnanamani, A., Senthilkumar, P. (2018). Surface active gold nanoparticles biosynthesis by new approach for bionanocatalytic activity. Journal of Photochemical Photobiological 179:119–125. DOI:

Verkerk, R.H.J. and Wright, D.J. (2003). Biological activity of neem seed kernel extract and synthetic azadirachtin against larvae of Plutella xylostellal. Pesticide science., 37,83-91. DOI:

Vijayaraghavan, K., Ashokkumar, T. (2017). Plant-mediated biosynthesis of metallic nanoparticles: a review of literature, factors affecting synthesis, characterization techniques and applications. Journal of Environmental Chemical Eng. 5:4866-4883. DOI:

Willner, B. and Basnar B (2007) Nanoparticle-enzyme hybrid systems for nanobiotechnology. FEBS Journal 274:302–309. DOI:

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