Phytochemical characterization and antifungal potentials of Melia azedarach Linn leave aqueous extract to inhibit aflatoxins biosynthesis in food during storages

Javid ALI; Arshad HUSSAIN; Muhammad SIDDIQUE; Muhammad AKRAM; Muhammad IKRAM; Muhammad ZAHOOR; Naila GULFAM; Raiz ULLAH; Amal ALOTAIBI

doi:10.15835/nbha53314558

Authors

Javid ALI Pakistan Council of Scientific and Industrial Research Laboratories Complex, Peshawar, 25120 (PK)
Arshad HUSSAIN Pakistan Council of Scientific and Industrial Research Laboratories Complex, Peshawar, 25120 (PK)
Muhammad SIDDIQUE Pakistan Council of Scientific and Industrial Research Laboratories Complex, Peshawar, 25120 (PK)
Muhammad AKRAM Pakistan Council of Scientific and Industrial Research Laboratories Complex, Peshawar, 25120 (PK)
Muhammad IKRAM Abdul Wali Khan University Mardan, Department of Chemistry, Mardan, Khyber Pakhtunkhwa, 23200 (PK)
Muhammad ZAHOOR University of Malakand at Chakdara, Department of Biochemistry, Dir Lower Khyber Pakhtunkhwa, 18800 (PK)
Naila GULFAM University of Peshawar Khyber, Jinnah College for Women, Department of Zoology, Pakhtunkhwa, 25120 (PK)
Raiz ULLAH King Saud University, MAPPRC, College of Pharmacy, Medicinal Aromatic and Poisonous Plants Research Center, Riyadh 11451 (SA)
Amal ALOTAIBI Princess Nourah bint Abdulrahman University, College of Medicine, Department of Basic Sciences, Riyadh 11671 (SA)

DOI:

https://doi.org/10.15835/nbha53314558

Keywords:

aflatoxins, flavonoids, mycelial growth inhibition, phenols, spore germination inhibition

Abstract

The current study was aimed to assess the chemical compositions of Melia azedarach Linn (Chinaberry) leaves aqueous extracts. Additionally, the extracts were also tested to investigate its antifungal potentials against Aspergillus flavus and Aspergillus parasiticus. Leaf extract of M. azedarach was obtained by maceration technique, subsequently analyzed using UV-Visible Spectrophotometer, Fourier Transform Infrared (FTIR) and Gas Chromatography-Mass Spectrometry (GC-MS). The total phenolic and flavonoids contents were; 67.5 ± 0.4 mg GAE/g DW and 12.7 ± 0.2 mg QE /g DW respectively. The presence phytochemicals were confirmed from various functional groups recorded in FT-IR spectra. The results were further validated through GC-MS analysis where a total of 18 compounds were identified with seven major compounds; namely 1-Butanol, 3-methyl-, acetate (11.53%), followed by coumaran (10.04%), (R, S)-2-propyl-5-oxohexanal (7.07%), 10-octadecenoic acid, methyl ester (5.16%) and 5,7-Octadien-2-one, 3-acetyl (3.06%). The extract exhibited antifungal activities against two major aflatoxin-producing fungi, A. flavus and A. parasiticus. The aqueous extract (31.25 to 500 mg mL^-1) was active to inhibit the spore germination, mycelial growth, biomass production and aflatoxin biosynthesis. Spore germination was significantly reduced, with maximum inhibition of 83% against A. flavus and 85% against A. parasiticus at 500 mg mL^-1. Mycelial growth and fungal biomass were markedly declined with increasing trend in extract concentration. The recorded biomass inhibition was 73.2% and 76.9% respectively against A. flavus and A. parasiticus. The extract also significantly suppressed the aflatoxin production in the selected fungal strains at higher concentrations, exceeded from 75% with respect to aflatoxins B₁, B₂, G₁, and G₂. The findings suggest that M. azedarach leaves extract is a valuable source of bioactive compounds possessing strong antifungal and anti-aflatoxigenic properties and could be considered as a promising natural alternative for controlling aflatoxins contamination in agricultural food sectors.

References

Abbas MK, Ahmad M, Barkat K, Aslam N (2017). Antifungal, antioxidant and phytochemical screening of Melia azedarach flower extracts by using different solvents. Journal of Pharmaceutical Research International 20(1):1-12. https://doi.org/10.9734/jpri/2017/38246

Ahmed MF, Ahmed MA, Thayyil H, Zameeruddin K, Ibrahim M (2008). Antioxidative activity of Melia azedarach Linn. leaf extract. Iranian Journal of Pharmacology & Therapeutics 7(1):31-34.

Ahmed OS, Tardif C, Rouger C, Atanasova V, Richard-Forget F, Waffo-Teguo P (2022). Naturally occurring phenolic compounds as promising antimycotoxin agents: Where are we now? Comprehensive Reviews in Food Science and Food Safety 21:1161-1197. https://doi.org/10.1111/1541-4337.12891

Akacha M, Lahbib K, Ghanem Boughanmi N (2016a). Phytochemical evaluation and net antioxidant activity of Melia azedarach L. leaves extracts from their ProAntidex parameter. Bangladesh Journal of Pharmacology 11:301-307. https://doi.org/10.3329/bjp.v11i2.25981

Akacha M, Lahbib K, Remadi MD, Ghanem N (2016b). Antibacterial, antifungal and anti-inflammatory activities of Melia azedarach ethanolic leaves extract. Bangladesh Journal of Pharmacology 11:666-674. https://doi.org/10.3329/bjp.v11i3.27000

Akpuaka A, Ekwenchi MM, Dashak DA, Dildar (2013). Biological activities of characterized isolated of n-hexane extract of Azadirachta indica a Juss (Neem) leaves. New York Science Journal 6(6):119-124. https://www.sciencepub.net/newyork/ny0606/

Aly A, Hussein E, Omar M, El-Abbasi I, Abd-Elsalam K (2011). Effect of fatty acid content on the level of cotton seed colonization by fungi. Biology Letters 48:125-137. https://doi.org/10.2478/v10120-011-0013-9

Aly SE, Sabry BA, Shaheen MS, Hathout AS (2016). Assessment of antimycotoxigenic and antioxidant activity of star anise (Illicium verum) in vitro. Journal of the Saudi Society of Agricultural Sciences 15:20-27. https://doi.org/10.1016/j.jssas.2014.05.003

Antara S, Amla B (2012). Evaluation of antimicrobial activity of different solvent extracts of medicinal plant: Melia azedarach L. International Journal of Current Pharmaceutical Research 4(2):67-73.

Asghar SF, Choudahry MI (2011). Gas chromatography-mass spectrometry (GC-MS) analysis of petroleum ether extract (oil) and bio-assays of crude extract of Iris germanica. International Journal of Genetics and Molecular Biology 3(7):95-100.

Behiry SI, Hamad NA, Alotibi FO, Al-Askar AA, Arishi AA, Kenawy AM, … Helfish AA (2022). Antifungal and antiaflatoxigenic activities of different plant extracts against Aspergillus flavus. Sustainability 14(19):12908. https://doi.org/10.3390/su141912908

Belakhdar G, Benjouad A, Abdennebi EHD (2015). Determination of some bioactive chemical constituents from Thesium humile Vahl. Journal of Materials and Environmental Science 6(10):2778-2783.

Cadenillas LF, Billerach, G, Hernandez C, Durrieu V, Bailly JD (2024). Inhibition of aflatoxin B1 production by procyanidins present in Annona muricata and Uncaria tomentosa aqueous extracts. Toxins 16(11):454. https://doi.org/10.3390/toxins16110454

Cadenillas LF, Hernandez C, Bailly S, Billerach G, Durrieu V, Bailly JD (2023a). Role of polyphenols from the aqueous extract of Aloysia citrodora in the inhibition of aflatoxin B1 synthesis in Aspergillus flavus. Molecules 28(13):5123. https://doi.org/10.3390/molecules28135123

Cadenillas LF, Hernandez C, Mathieu C, Bailly JD, Durrieu V (2023b). Screening of the anti-aflatoxin B1 activity of Peruvian plant extracts: relation with their composition. Food and Bioprocess Technology 16:1324-1334. https://doi.org/10.1007/s11947-023-03002-7

Carpinella MC, Ferrayoli GC (2005). Antifungal synergistic effect of scopoletin, a hydroxycoumarin isolated from Melia azedarach L. fruits. Journal of Agricultural and Food Chemistry 53:2922-2927. https://doi.org/10.1021/jf0482461

Carpinella MC, Giorda LM, Ferrayoli GC (2003). Antifungal effects of different organic extracts from Melia azedarach L. on phytopathogenic fungi and their isolated active components. Journal of Agricultural and Food Chemistry 51:2506-2511. https://doi.org/10.1021/jf026083f

Charoensin S (2014). Antioxidant and anticancer activities of Moringa oleifera leaves. Journal of Medicinal Plants Research 8(7):318-325. https://doi.org/10.5897/jmpr2013.5353

Cho KH, Hong JH, Lee KT (2010). Monoacylglycerol (MAG)-oleic acid has stronger antioxidant, anti-atherosclerotic, and protein glycation inhibitory activities than MAG-palmitic acid. Journal of Medicinal Food 13:99-107. https://doi.org/10.1089/jmf.2009.1024

Desbois AP, Smith VJ (2010). Antibacterial free fatty acids: Activities, mechanisms of action and biotechnological potential. Applied Microbiology and Biotechnology 85:1629-42. https://doi.org/10.1007/s00253-009-2355-3

Ezhilan BP, Neelamegam R (2012). GC-MS analysis of phytocomponents in the ethanol extract of Polygonum chinense L. Pharmacognosy Research 4(1):11-14. https://doi.org/10.4103/0974-8490.91028

Ganesan T, Subban M, Christopher Leslee DB, Kuppannan SB, Seedevi P (2024). Structural characterization of n-hexadecanoic acid from the leaves of Ipomoea eriocarpa and its antioxidant and antibacterial activities. Biomass Conversion and Biorefinery 14:14547-14558. https://doi.org/10.1007/s13399-022-03576-w

Gopalakrishnan K, Udayakumar R (2014). GC-MS analysis of phytocompounds of leaf and stem of Marsilea quadrifolia (L.). International Journal of Biochemistry Research & Review 4:517-526. https://doi.org/10.9734/ijbcrr/2014/11350

Gurning K, Iksen I., Simanjuntak HA, Purba H (2020). Identification of the chemical compound of essential oil from Ketumbar (Coriandrum sativum L.) leaves with GC-MS. Pharmacognosy Journal 12(5):1019-23. https://doi.org/10.5530/pj.2020.12.144

Harini V, Kumar PR, Thirumal M (2022). Phytoconstituents screening, TLC, and GC-MS analysis of Barleria Cristata Linn. leaves methanolic extract. Journal of Pharmaceutical Negative Results 13(8):4445-4450. https://doi.org/10.47750/pnr.2022.13.s08.569

Hernandez C, Cadenillas L, Maghubi AE, Caceres I, Durrieu V, Mathieu C, Bailly JD (2021). Mimosa tenuiflora aqueous extract: role of condensed tannins in anti-aflatoxin B1 activity in Aspergillus flavus. Toxins 13(6):391. https://doi.org/10.3390/toxins13060391

Hrichi S, Chaâbane-Banaoues R, Alibrando F, Altemimi AB, Babba O, Majdoub YOE, … Cacciola F (2022). Chemical composition, antifungal and anti-biofilm activities of volatile fractions of Convolvulus althaeoides L. roots from Tunisia. Molecules 27(20):6834. https://doi.org/10.3390/molecules27206834

Izzah FZ, Sitti RAH, Nur AM (2021). Optimisation of extraction method and phytochemical compounds of green Christia vespertilionis Leaves using GC-MS. International Journal of Pharmaceutical Sciences and Review and Research 70(1):1-8. http://doi.org/10.47583/ijpsrr.2021.v70i01.001

Jaafar NS, Hamad MN, Abbas IS, Jaafar IS (2016). Qualitative phytochemical comparison between flavonoids and phenolic acids contents of leaves and fruits of Melia Azedirach (Family: Meliaceae) cultivated in Iraq By HPLC and HPTLC. International Journal of Pharmacy and Pharmaceutical Sciences 8(10):242-250. https://doi.org/10.22159/ijpps.2016v8i10.13868

Jabeen K, Javaid A, Ahmad E, Athar M (2011). Antifungal compounds from Melia azedarach leaves for management of Ascochyta rabiei, the cause of chickpea blight. Natural Product Research 25(3):264-276. https://doi.org/10.1080/14786411003754298

Jabeen K, Javaid A, Athar M (2008). Fungistatic activity of aqueous and organic solvent extracts of Melia azedarach against Ascochyta rabiei. Pakistan Journal of Phytopathology 20:143-149.

Jahirhussain G, Kala K, Ayyappan P, Muniappan V, Tamilselvan V, Rajkumar P (2015). Profiling the secondary chemical class in vivo Melia composite wild leaf using ethanolic fraction. World Journal of Pharmaceutical Research 14(12):1367-1376.

Javaid A, Rehman HA (2011). Antifungal activity of leaf extracts of some medicinal trees against Macrophomina phaseolina. Journal of Medicinal Plants Research 5(13):2868-2872.

Javid S, Shanmugarajan D, Kumar HY, Arivuselvam R, Anjum NF, Purohit MN, ... Kumar (2024). Rational design, synthesis, analysis and antifungal activity of novel myristic acid derivatives as N-myristoyltransferase inhibitors. Journal of Molecular Structure 1303:137568. https://doi.org/10.1016/j.molstruc.2024.137568

Kumari SA, Kumar V, Guleria I, Sharma M, KumarA, Alruways MW, Khan N, Raina (2021). Antimicrobial potential and chemical profiling of leaves essential oil of Mentha species growing under north-west Himalaya conditions. Journal of Pure and Applied Microbiology 15(4):2229-2243. https://doi.org/10.22207/jpam.15.4.45

Karthikeyan M, Subramanian P, Ramalingam S (2019). Phytochemical analysis in economically important Ficus benghalensis L. and Ficus krishnae C. DC. using GC-MS. International Journal of Pharmaceutical and Biological Science 10:5-13. https://doi.org/10.22376/ijpbs.2019.10.4.p5-13

Krishnaveni M, Dhanalakshmi R, Nandhini N (2014). GC-MS analysis of phytochemicals, fatty acid profile, antimicrobial activity of Gossypium seeds. International Journal of Pharmaceutical Sciences Review and Research 27:273-276.

Liu S, Ruan W, Li J, Xu H, Wang J, Gao Y, Wang J (2008). Biological control of phytopathogenic fungi by fatty acids. Mycopathologia 166:93-102. https://doi.org/10.1007/s11046-008-9124-1

Loran S, Carraminana JJ, Juan T, Arino A, Herrera M (2022). Inhibition of Aspergillus Parasiticus growth and aflatoxins production by natural essential oils and phenolic acids. Toxins 14(6):384. https://doi.org/10.3390/toxins14060384

Mickymaray S, Al Aboody MS, Rath PK, Annamalai P, Nooruddin T (2016). Screening and antibacterial efficacy of selected Indian medicinal plants. Asian Pacific Journal of Tropical Biomedicine 6(3):185-191. https://doi.org/10.1016/j.apjtb.2015.12.005

Mpofu A, Sapirstein HD, Beta T (2006). Genotype and environmental variation in phenolic content, phenolic acid composition, and antioxidant activity of hard spring wheat. Journal of Agriculture and Food Chemistry 54(4):1265-70. https://doi.org/10.1021/jf052683d

Mujeeb F, Bajpai P, Pathak N (2014). Phytochemical evaluation, antimicrobial activity, and determination of bioactive components from leaves of Aegle marmelos. BioMed Research International 2014(1):497606. http://dx.doi.org/10.1155/2014/497606

Munir A, Sultana B, Babar T, Bashir A, Amjad M, Hassan Q (2012). Investigation on the antioxidant activity of leaves, fruit and stem bark of Dhraik (Melia azedarach). European Journal of Applied Sciences 4(2): 47-51.

Mwamatope B, Tembo D, Chikowe I, Kampira E, Nyirenda C (2020). Total phenolic contents and antioxidant activity of Senna singueana, Melia azedarach, Moringa oleifera and Lannea discolor herbal plants. Scientific Africa 9:e00481. https://doi.org/10.1016/j.sciaf.2020.e00481

Neycee MA, Nematzadeh GHA, Dehestani, Alavi AM (2012). Assessment of antifungal effects of shoot extracts in chinaberry (Melia azedarach) against 5 phytopathogenic fungi. International Journal of Agriculture and Crop Sciences 4(8):474-477.

Omoruyi BE, Afolayan AJ, Bradley G (2014). Chemical composition profiling and antifungal activity of the essential oil and plant extracts of Mesembryanthemum edule (L.) bolus leaves. African Journal of Traditional, Complementary and Alternative Medicines 11(4):19-30. https://doi.org/10.4314/ajtcam.v11i4.4

Ponzilacqua B, Corassin CH, Oliveira CAF (2018). Antifungal activity and detoxification of aflatoxins by plant extracts: Potential for food applications. The Open Food Science Journal 10(1):24-32.

Rajashekar Y, Raghavendra A, Bakthavatsalam N (2014). Acetylcholinesterase inhibition by biofumigant (Coumaran) from leaves of Lantana camara in stored grain and household insect pests. BioMed Research International 2014(1):187019. https://doi.org/10.1155/2014/187019

Rouis-Soussi LS, Boughelleb-M'Hamdi N, El Ayeb-Zakhama A, Flamini G, Ben Jannet H, Harzallah-Skhiri F (2014). Phytochemicals, antioxidant and antifungal activities of Allium roseum var. grandiflorum subvar. typicum Regel. South African Journal of Botany 91:63-70. https://doi.org/10.1016/j.sajb.2013.12.005

Sana N, Javiad A, Shoaib A (2017). Antifungal activity of methanolic leaf extracts of allelopathic trees against Sclerotium rolfsii Sacc. Bangladesh Journal of Botany 46(3):987-993.

Sen A, Batra A. (2012). Evaluation of antimicrobial activity of different solvent extracts of medicinal plant: Melia azedarach L. International Journal of Current Pharmaceutical Research 4(2):67-73.

Shaaban MT, Ghaly MF, Fahmi SM (2021). Antibacterial activities of hexadecanoic acid methyl ester and green-synthesized silver nanoparticles against multidrug-resistant bacteria. Journal of Basic Microbiology 61:557-68. https://doi.org/10.1002/jobm.202100061

Shrestha SS, Ferrarese I, Sut S, Zengin G, Grana S, Ak G, Pant DR, Dall'Acqua S, Rajbhandary S (2021). Phytochemical investigations and in vitro bioactivity screening on Melia azedarach L. leaves extract from Nepal. Chemistry and Biodiversity 18(5):e2001070. https://doi.org/10.1002/cbdv.202001070

Sownthariya C, Shanthi P (2022). Spectroscopic analysis (GC-MS and FT-IR) of Melia azedarach leaf extract. International Journal of Botany Studies 7(2):133-141.

Sreelatha S, Padma PR (2009). Antioxidant activity and total phenolic content of Moringa oleifera leaves in two stages of maturity. Plant Foods for Human Nutrition 64:303-311. https://doi.org/10.1007/s11130-009-0141-0

Sung WS, Lee DG (2010). Antifungal action of chlorogenic acid against pathogenic fungi, mediated by membrane disruption. Pure and Applied Chemistry 82:219-226. https://doi.org/10.1351/pac-con-09-01-08

Swamy MK, Sinniah UR, Akhtar MS (2015). In vitro pharmacological activities and GC-MS analysis of different solvent extracts of Lantana camara leaves collected from tropical region of Malaysia. Evidence Based Complementary and Alternative Medicine 2015:506413. https://doi.org/10.1155/2015/506413

Tian F, Woo SY, Lee SY, Park SB, Im JH, Chun HS (2023). Plant-based natural flavonoids show strong inhibition of aflatoxin production and related gene expressions correlated with chemical structure. Food Microbiology 109:104141. https://doi.org/10.1016/j.fm.2022.104141

US EPA (2015). United Sates Environmental Protection Agency, Washington, DC. Office of Prevention Pesticides and Toxic Substances pp 1-15.

Velazhahan R, Vijayanandraj S, Vijayasamundeeswari A, Paranidharan V, Samiyappan R, Iwamoto T, Muthukrishnan, S (2010). Detoxification of aflatoxins by seed extracts of the medicinal plant, Trachyspermum ammi (L.) Sprague ex Turrill - Structural analysis and biological toxicity of degradation product of aflatoxin G1. Food Control 21(5):719-725. https://doi.org/10.1016/j.foodcont.2009.10.014

Venn-Watson S, Schork N (2023). Pentadecanoic acid (C15:0), an essential fatty acid, shares clinically relevant cell-based activities with leading longevity-enhancing compounds. Nutrients 15(21):4607. https://doi.org/10.3390/nu15214607

Vyas S, Kachhwaha S, Kothari SL (2015). Comparative analysis of phenolic contents and total antioxidant capacity of Moringa oleifera Lam. Pharmacognosy Journal 7(1): 44-51.

Yazdani D, Mior Ahmad ZA, Yee How T, Jaganath IB, Shahnazi S (2013). Inhibition of aflatoxin biosynthesis in Aspergillus flavus by phenolic compounds extracted of Piper betle L. Iranian Journal of Microbiology 5(4):428-433.

Youssef NH, Qari SH, Behiry SI, Dessoky ES, El-Hallous EI, Elshaer MM, … Heflish AA (2021). Antimycotoxigenic activity of beetroot extracts against Altenaria alternata mycotoxins on potato crop. Applied Sciences 11(9):4239. https://doi.org/10.3390/app11094239

Zahija I, Jersek B, Demsar L, Polak ML, Polak T (2023). Production of aflatoxin B1 by Aspergillus parasiticus grown on a novel meat-based. Media. Toxins 15(1):25. https://doi.org/10.3390/toxins15010025

Zishan M, Dhar DW, Manzoor U (2024). Fourier transform infrared spectroscopy analysis and phytochemical screening of selected medicinal plant extracts. African Journal of Biological Sciences 6(7):4021-4029.

Zhou W, Hu LB, Zhao Y, Wang MY, Zhang H, Mo HZ (2015). Inhibition of fungal aflatoxin B1 biosynthesis by diverse botanically-derived polyphenols. Tropical Journal of Pharmaceutical Research 14(4):605-609. https://doi.org/10.4314/tjpr.v14i4.7