Cytogenotoxicity and safe doses of Azadirachta indica L. (Neem tree) leaf extract obtained in Birnin Kebbi, Nigeria

Tajudeen Yahaya, Salisu Titilola Fausat, Angela Daniel, Muhammed Ndakogi Musa, Emmanuel John, Ridwan Sulaiman, Abubakar Saadu, ThankGod James

Abstract


Azadirachta indica (neem tree) is used to treat diseases in Nigeria, however, scarcity of documented information on the plant's toxicity exists in the country. Toxicity evaluation of medicinal plants in relation to geographical location is essential, as toxicity may vary due to differences in pollutants and environmental factors. This study assessed the phytochemical composition, heavy metal content, cytotoxic potential, and safe doses of methanolic leaf extracts of A. indica obtained from Birnin Kebbi, Nigeria. Twenty-one Allium cepa bulbs were divided equally into seven groups. The negative control (Group 1) and positive control (Group 2) bulbs were grown for five days over beakers containing distilled water and formaldehyde, respectively. Test groups (Groups 3–7) were grown over beakers containing A. indica extract solutions at concentrations of 0.25, 0.5, 1.0, 2.0, and 4.0 g, respectively. Phytochemical analysis revealed abundant flavonoids, phenols, and saponins, moderate levels of tannins, and trace amounts of quinones, cardiac glycosides, and terpenoids. Heavy metal analysis showed the presence of copper (3.8 mg/kg), lead (0.12 mg/kg), and cadmium (0.04 mg/kg), all of which exceeded the permissible limits established by WHO. Except for the 0.25 g group, the test groups exhibited dose-dependent (p ≤ 0.05) cytogenetic effects, including reduced root growth and mitotic index. Chromosomal abnormalities such as sticky, bridged, vagrant, laggard, and fragmented chromosomes were also observed. These findings suggest that A. indica may be safe at low concentrations but exhibits toxic effects at higher concentrations. The results provide a valuable baseline for further investigations into the plant's safety.

Keywords: Azadirachta indica (neem tree), lead, mitotic index, saponins, vagrant chromosomes.

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References


Adegbite AE, Ayodele MS, Odunbaku KR, Idehen EO. 2009. Mutagenic effect of neem leaf extract used in traditional medicine on Allium cepa (L.). Scientific Research and Essays 4(11): 1315-1321. https://doi.org/10.5897/SRE.9000670.

Ajai AI, Ochigbo SS, Shaba EY, Mohammmed H. 2014. Determination of heavy metal profiles in Neem leaves (Azadirachta indica A) along some major streets in Minna metropolis Nigeria. Journal of Natural Science Research 4(3): 7-12.

http://www.iiste.org/Journals/index.php/JNSR/article/view/11059/11360.

Akan CJ, Inuwa LB, Chellube ZM, Lawan B. 2013. Heavy metals in leaf, stem bark of neem tree (Azadirachta indica) and roadside dust in Maiduguri Metropolis, Borno State, Nigeria. Environment and Pollution 2(1): 88-95. https://doi.org/10.5539/ep.v2n1p88.

Akaneme FI, Amaefule CC. 2012. Evaluation of the cytotoxicity and genotoxicity of aqueous leaf extracts of Azadirachta indica A. Juss using the Allium test. Journal of Medicinal Plants Research 6(22): 3898-3907. https://doi.org/10.5897/JMPR12.427.

Alzohairy MA. 2016. Therapeutics role of Azadirachta indica (Neem) and their active constituents in diseases prevention and treatment. Evidence-based complementary and alternative medicine Article ID 7382506. https://doi.org/10.1155/2016/7382506.

Bankole SO, Alao DO, Ariwoola OS, Osunlaja OA, Oladejo SF, Chukwudebe E. 2018. Investigation of Heavy Metal Status in Neem Tree (Azadiractha Indica) Barks Existing Along Some Selected Major Roads in Ibadan, Oyo State. International Journal of Agriculture, Environment and Bioresearch 3 (4): 515-522.

https://ijaeb.org/uploads2018/AEB_03_230.pdf.

Blum FC, Singh J, Merrell DS. 2019. In vitro activity of neem (Azadirachta indica) oil extract against Helicobacter pylori. Journal of Entomopharmacology 232: 236–243.

https://doi.org/10.1016/j.jep.2018.12.025.

Bolaji O, Abolade YA, Aduwa S, Isiaka AB, Durodola O, Adeoye A. 2024. Potential health and environmental benefits of the identified phytochemicals screening of (Azadirachta indica) neem leaves in Bauchi Metropolis, Bauchi State, Nigeria. GSC Biological and Pharmaceutical Sciences 26(03): 068–083.

https://doi.org/10.30574/gscbps.2024.26.3.0037.

Fasae OA, Aganto TO. 2024. Chemical composition of neem (Azadiractha indica) plant parts as feed resources for ruminants. Nigerian Journal of Animal Production 1: 742–745.

https://doi.org/10.51791/njap.vi.5780.

Galati G, O'Brien PJ. 2004. Potential toxicity of flavonoids and other dietary phenolics: significance for their chemopreventive and anticancer properties. Free Radical Biology and Medicine 37(3): 287–303. https://doi.org/10.1016/j.freeradbiomed.2004.04.034.

Giannakoula A, Therios I, Chatzissavvidis C. 2021. Effect of Lead and Copper on photosynthetic apparatus in citrus (Citrus aurantium L.) plants. The role of antioxidants in oxidative damage as a response to heavy metal stress. Plants (Basel, Switzerland) 10(1): 155. https://doi.org/10.3390/plants10010155.

Islas JF, Acosta E, G-Buentelloa Z, Delgado-Gallegosa JL, Moreno-Treviñoc GM, Escalantec B, Moreno-Cuevasc JE. 2020. An overview of Neem (Azadirachta indica) and its potential impact on health. Journal of Functional Foods 74: 104-171.

https://doi.org/10.1016/j.jff.2020.104171.

Keta JN, Suberu HA, Shehu K, Yahayya U, Mohammad NK, Gudu GB. 2019. Effect of neem (Azadirachta indica A. Juss) leaf extract on the growth of Aspergillus species isolated from foliar diseases of rice (Oryza Sativa). Science World Journal 14(1): 98-102.

https://www.ajol.info/index.php/swj/article/view/208446.

Khalid S, Shahid M, Shah AH, Saeed F, Ali M, Dumat C. 2020. Heavy metal contamination and exposure risk assessment via drinking groundwater in Vehari, Pakistan. Environmental Science and Pollution Research 27: 39852-39864. https://doi.org/10.1007/s11356-020-10106-6.

Khanal S. 2021. Qualitative and quantitative phytochemical screening of Azadirachta indica (Juss.) plant parts. International Journal of Applied Science and Biotechnology 9(2): 122-127. https://doi.org/10.3126/ijasbt.v9i2.38050.

Kilani-Morakchi S, Morakchi-Goudjil H, Sifi K. 2021. Azadirachtin-based insecticide: Overview, risk assessments, and future directions. Frontiers in Agronomy 3: 676208; https://doi.org/10.3389/fagro.2021.676208.

Li H, Tsao R, Deng Z. 2012. Factors affecting the antioxidant potential and health benefits of plant foods. Canadian Journal of Plant Science 92 (6): 1101-1111.

https://doi.org/10.4141/CJPS2011-239.

Maji S, Modak S. 2021. Neem: Treasure of natural phytochemicals. Chemical Science Review and Letters 10(39): 396-401. https://doi.org/10.37273/chesci.cs205205351.

Marrelli M, Conforti F, Araniti F, Statti GA. 2016. Effects of Saponins on lipid metabolism: A review of potential health benefits in the treatment of obesity. Molecules (Basel, Switzerland) 21(10):1404. https://doi.org/10.3390/molecules21101404.

Mishra A, Dave N. 2013. Neem oil poisoning: Case report of an adult with toxic encephalopathy. Indian Journal of Critical Care Medicine 17(5): 321–322.

https://doi.org/10.4103/0972-5229.120330.

Mofio BM, Umar ID, Mmadu’abuchi UM, Isah MC, Nyadar PM. 2024. Phytochemical properties and nutritional status of Azadirachta indica leaf ethanolic extract from Iddo Sarki in FCT, Abuja Nigeria. Direct Research Journal of Public Health and Environmental Technology 9(2): 192-198. https://doi.org/10.26765/DRJPHET012714757.

New Jersey Department of Health. 2015. Hazardous substances fact sheet: Phenols. Available at https://nj.gov/health/eoh/rtkweb/documents/fs/1487.pdf (Accessed March 1, 2022).

Nyamai DW, Arika W, Ogola PE, Njagi ENM, Ngugi MP. 2016. Medicinally important phytochemicals: An untapped research avenue. Journal of Pharmacognosy and Phytochemistry 4(1):35-49. https://www.rroij.com/open-access/medicinally-important-phytochemicals-an-untapped-research-avenue-.pdf.

Obadahun J, Okeh Q, Shittu KJ, Pascalina PL, Oludege OF, Omotola MB. 2020. Assessment of heavy metals of some selected medicinal plant in Kwakwachi Fagge Local Government of Kano State, Nigeria. International Journal of Research and Scientific Innovation 7(8): 80-82. https://ideas.repec.org/a/bjc/journl/v7y2020i8p80-82.html.

Ogbuewu IP, Odoemenam HO, Obikaonu MN, Opara OO, Emenalom MC, Uchegbu IC, Okoli BO, Iloeje MU. 2011. The growing importance of neem (Azadirachta indica A. Juss) in agriculture, industry, medicine and environment: A Review. Research Journal of Medicinal Plants 5:230-245. https://doi.org/10.3923/rjmp.2011.230.245.

Onimisi BO, Nadabo YA, Musa SA, Musa MA. 2015. Nephrotoxic effects of methanolic extract of Azadirachta indica (Neem) root bark on the histology of adult Wistar rats. International Journal of Health Sciences & Research 5 (4): 116-121.

Rahimzadeh FM, Rahimzadeh M, Kazemi S, Moghadamnia AA. 2017. Cadmium toxicity and treatment: An update. Caspian Journal of Internal Medicine 8(3):135–145.

https://doi.org/10.22088/cjim.8.3.135.

Samtiya M, Aluko RE, Dhewa T. 2020. Plant food anti-nutritional factors and their reduction strategies: an overview. Food Production Process and Nutrition 2(6):23-35.

https://doi.org/10.1186/s43014-020-0020-5.

Seriana I, Akmal M, Darusman D, Wahyuni S, Khairan K, Sugito S. 2021. Neem leaf (azadirachta indica a. juss) ethanolic extract on the liver and kidney function of rats. The Scientific World Journal 5:1-7. https://doi.org/10.1155/2021/7970424.

Skibola CF, Smith MT. 2000. Potential health impacts of excessive flavonoid intake. Free Radical Biology Medicine August 29(3-4): 375-83. https://doi.org/10.1016/s0891-5849(00)00304-x.

Soliman IM. 2001. Genotoxicity testing of Neem Plant (Azadirachta indica A. Juss.) using the Allium cepa chromosome aberration assay. Journal of Biological Sciences 1(11): 1021-1027. https://doi.org/10.3923/jbs.2001.1021.1027.

Sulayman AF, Abdu H, Saidu AA, Ahmad F, Suleiman JZ, Isyaku S. 2021. Assessment of atmospheric metal pollution biomonitoring potential by Neem Tree (Azadirachta indica) leaves in Katsina metropolitan area of Katsina State, Nigeria. Current Approaches in Science and Technology Research 3:37–45. https://doi.org/10.9734/bpi/castr/v3/8345D.

Taylor AA, Tsuji JS, Garry MR. 2020. Critical review of exposure and effects: Implications for setting Regulatory Health Criteria for ingested Copper. Environmental Management 65:131–159. https://doi.org/10.1007/s00267-019-01234-y.

Tibebu A, Haile G, Kebede A. 2018. Review on medicinal value and other application of Neem Tree: Senior Seminar on Animal Health. Biomedicine and Nursing 4(1): 62-69.

https://www.arcjournals.org/pdfs/ajiv/v2-i2/4.pdf.

Ujah II, Nsude CA, Ani ON, Alozieuwa UB, Okpako IO, Okwor AE. 2021. Phytochemicals of neem plant (Azadirachta indica) explains its use in traditional medicine and pest control. GSC Biological and Pharmaceutical Sciences 14(2):165–171.

https://doi.org/10.30574/gscbps.2021.14.2.0394.

United State Center for Disease Control. 2021. Health problems caused by Lead. Available at https://www.cdc.gov/niosh/topics/lead/health.html# (Accessed March 2, 2022).

United States Environmental Protection Agency. 2022. Phenols: Health Hazards. Available at https://www.epa.gov/sites/default/files/2016-09/documents/phenol.pdf (Accessed March 1, 2022).

Uwague A. 2019. Comparative potential qualitative and quantitative phytochemical evaluation of Neem and Moringa Oleifera leaf plants in Ozoro, Delta State, Nigeria. International Scientific Research in Science and Technology 6:120-340.

https://doi.org/10.32628/ijsrst196652.

Virshette SJ, Patil M, Deshmukh AA, Shaikh JR. 2019. Phytochemical Analysis of Different Extract of Azadirachta indica Leaves. International Journal of Pharmacological Science Reviewed Research 59(1):161-165. https://globalresearchonline.net/journalcontents/v59-1/27.pdf.

World Health Organization. 2020. Safety evaluation of certain food additives and contaminants: supplement 2: pyrrolizidine alkaloids, prepared by the eightieth meeting of the Joint FAO/WHO Expert Committee on Food Additives (‎‎JECFA) [Internet]. Available at https://www.who.int/publications/i/item/9789240012677 (Accessed June 27, 2023).

Yahaya T, Okpuzor J, Oladele E. 2012. Investigation of cytotoxicity and mutagenicity of cement dust using Allium cepa test. Research Journal of Mutagen 2(1):10-18.

https://doi.org/10.3923/rjmutag.2012.10.18.

Yahaya T, Esther O, Mutiu S, Aminu M, Obijaku E. 2021. Characterization and cytogenotoxicity of water samples from River Challawa in Kano, Northwest Nigeria. Songklanakarin Journal of Science and Technology 43(4):1-8.

https://sjst.psu.ac.th/journal/43-4/6.pdf.

Yahaya OT, Oladele EO, Bunza MDA, Yusuf AB, Izuafa A, Danjuma JB. 2022. Hematotoxicity and nephrotoxicity of long-term administration of Guiera senegalensis (J.F. Gme), Cassia occidentalis (Linn), and Ziziphus mauritiana (Lam) leaves obtained in Birnin Kebbi, Nigeria. Journal of Herbmed Pharmacology 11(3):367-374.

https://doi.org/10.34172/jhp.2022.42.

Yarmohammadi F, Mehri S, Najafi N, Amoli S, Hosseinzadeh H. 2021. The protective effect of Azadirachta indica (neem) against metabolic syndrome: A review. Iranian Journal of Basic Medical Sciences 24(3):280-292. https://doi.org/10.22038/ijbms.2021.48965.11218


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