Investigation of the Effects of Ganoderic Acid and Copper Nanoparticles on Aflatoxin B1 Production by Aspergillus flavus

Subject Areas : Mycology

Mohaddeseh Larypoor ¹ , Elaheh Sabori ² , Tina Zarrinpanah ³ , Hamidreza Shirmohammadi ⁴ , Fereshteh Rahmati ⁵

1 - , Tehran, IranDepatment of Microbiology, NT.C., Islamic Azad University
2 - Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran
3 - Department of Microbiology, Islamic Azad University, Kish International Branch, Kish, Iran.
4 - Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran
5 - Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran

Received: 2025-02-24 Accepted : 2025-05-19 Published : 2025-06-10

Keywords: Aflatoxin B1, Aspergillus flavus, ganoderic acid, copper nanoparticles, antifungal activity.,

Abstract :

Background and Objective: Aflatoxin B1 is one of the most critical mycotoxins that enters the human body through the consumption of food and has carcinogenic effects in humans. Edible and medicinal macrofungi contain active compounds with antimicrobial and anticancer properties.The objective of this study was to investigate the inhibitory and antitoxigenic effects of ganoderic acid derived from Ganoderma lucidum and a copper nanoparticle (CuNP), both individually and synergistically, on the growth of Aspergillus flavus and the production of Aflatoxin B1.

Materials and Methods: I In this experimental research, ganoderic acid and the copper nanoparticle were extracted and prepared using standard methods. Antifungal activity was evaluated using the disk diffusion assay, Minimum Inhibitory Concentration (MIC), Minimum Fungicidal Concentration (MFC), and Inhibitory Concentration 50% (IC₅₀) tests. High-Performance Liquid Chromatography (HPLC) was used to measure the level of Aflatoxin B1.

Results: Both compounds effectively inhibited fungal growth. The copper nanoparticle completely inhibited growth at a concentration of 125 µg/ml (MIC), while ganoderic acid achieved this effect at a concentration of 250 µg/ml. However, HPLC results indicated that ganoderic acid significantly reduced Aflatoxin B1 more than the other treatments

Conclusion: Ganoderic acid can be proposed as a safe, effective, and biological compound for inhibiting Aspergillus flavus and reducing Aflatoxin B1 levels in foodstuffs and animal feed. The findings suggest that the addition of the copper nanoparticle to this compound did not significantly improve the antitoxigenic performance, and future research should focus on a more precise investigation of the molecular mechanisms of these interactions.

References:

1. S.Z. Iqbal, Mycotoxins in food, recent development in food analysis and future challenges; a review, Current Opinion in Food Science, 42 (2021) 237–247.
2. P. Pinton, C. Terciolo, D. Payros, I.P. Oswald, Mycotoxins hazard: the European view, Current Opinion in Food Science, DOI (2025) 101306.
3. G.S. Shephard, Aflatoxin analysis at the beginning of the twenty-first century, Analytical and bioanalytical Chemistry, 395 (2009) 1215–1224.
4. M. Sayadi, H. Tajik, Detoxification of aflatoxin b1 by Lactobacillus rhamnosus in a simulated model of the human digestive system, Journal of Microbial World, 11 (2019) 380–391 (In Persian).
5. D.K. Mahato, K.E. Lee, M. Kamle, S. Devi, K.N. Dewangan, P. Kumar, S.G. Kang, Aflatoxins in food and feed: An overview on prevalence, detection and control strategies, Frontiers in microbiology, 10 (2019) 2266.
6. N. Benkerroum, A. Ismail, Human breast milk contamination with aflatoxins, impact on children’s health, and possible control means: A review, International Journal of Environmental Research and Public Health, 19 (2022) 16792.
7. U.P. Sarma, P.J. Bhetaria, P. Devi, A. Varma, Aflatoxins: implications on health, Indian Journal of Clinical Biochemistry, 32 (2017) 124–133.
8. Y. Liu, C.-C.H. Chang, G.M. Marsh, F. Wu, Population attributable risk of aflatoxin-related liver cancer: systematic review and meta-analysis, European journal of cancer, 48 (2012) 2125–2136.
9. C. Wild, J. Miller, J. Groopman, Effects of aflatoxins on aflatoxicosis and liver cancer, Mycotoxin Control in Low-and Middle-Income Countries. International Agency for Research on Cancer, DOI (2015).
10. K. Muaz, M. Riaz, C.A.F.d. Oliveira, S. Akhtar, S.W. Ali, H. Nadeem, S. Park, B. Balasubramanian, Aflatoxin M1 in milk and dairy products: Global occurrence and potential decontamination strategies, Toxin reviews, 41 (2022) 588–605.
11. P. Chandra, Aflatoxins: Food safety, human health hazards and their prevention, Aflatoxins-Occurrence, Detoxification, Determination and Health Risks, IntechOpen2021.
12. W.H. Organization, Safety evaluation of certain contaminants in food: prepared by the eighty-fourth meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), Safety evaluation of certain contaminants in food: prepared by the eighty-fourth meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA), 2019.
13. M. Mohammadi, S.J. Hashemi, S. Rezaei, M. Bayat, Assessment of antifungal activity of Rosemary oil extract and its effect on AFL1 gene expression in Aspergillus flavus by Real-Time PCR, Journal of Microbial World, 11 (2018) 88–100 (In Persian).
14. M. Piotrowska, Microbiological decontamination of mycotoxins: opportunities and limitations, Toxins, 13 (2021) 819.
15. M.A. Klich, Aspergillus flavus: the major producer of aflatoxin, Molecular plant pathology, 8 (2007) 713–722.
16. A.M. Fouad, D. Ruan, H.K. El-Senousey, W. Chen, S. Jiang, C. Zheng, Harmful effects and control strategies of aflatoxin b1 produced by Aspergillus flavus and Aspergillus parasiticus strains on poultry, Toxins, 11 (2019) 176.
17. Sayadi, M., & Tajik, H. (2019). Detoxification of aflatoxin b1 by Lactobacillus rhamnosus in a simulated model of the human digestive system. Journal of Microbial World, 11(37), 380–391 (In Persian).
18. Q. Li, X. Zhu, Vanillin and its derivatives, potential promising antifungal agents, inhibit Aspergillus flavus spores via destroying the integrity of cell membrane rather than cell wall, Grain & oil science and technology, 4 (2021) 54–61.
19. Z. Peng, L. Chen, Y. Zhu, Y. Huang, X. Hu, Q. Wu, A.K. Nüssler, L. Liu, W. Yang, Current major degradation methods for aflatoxins: A review, Trends in food science & technology, 80 (2018) 155–166.
20. S. Gibellato, L. Dalsóquio, I. Do Nascimento, T. Alvarez, Current and promising strategies to prevent and reduce aflatoxin contamination in grains and food matrices, World Mycotoxin Journal, 14 (2021) 293–304.
21. B.L. Gonçalves, K. Muaz, C.F.S.C. Coppa, R.E. Rosim, E.S. Kamimura, C.A.F. Oliveira, C.H. Corassin, Aflatoxin M1 absorption by non-viable cells of lactic acid bacteria and Saccharomyces cerevisiae strains in Frescal cheese, Food research international, 136 (2020) 109604.
22. M. Kozarski, A. Klaus, D. Jakovljević, N. Todorović, W.A.A.Q.I. Wan, M. Nikšić, Ganoderma lucidum as a cosmeceutical: Antiradical potential and inhibitory effect on hyperpigmentation and skin extracellular matrix degradation enzymes, Archives of biological sciences, 71 (2019) 253–264.
23. K. Hapuarachchi, T. Wen, C. Deng, J. Kang, K. Hyde, Mycosphere essays 1: Taxonomic confusion in the Ganoderma lucidum species complex, Mycosphere, 6 (2015) 542–559.
24. T. Kohno, T. Hai-Bang, Q. Zhu, Y. Amen, S. Sakamoto, H. Tanaka, S. Morimoto, K. Shimizu, Tubulin polymerization-stimulating activity of Ganoderma triterpenoids, Journal of natural medicines, 71 (2017) 457–462.
25. F.F. Radwan, J.M. Perez, A. Haque, Apoptotic and immune restoration effects of ganoderic acids define a new prospective for complementary treatment of cancer, Journal of clinical & cellular immunology, DOI (2011) 004.
26. B.S. Gill, Navgeet, S. Kumar, Ganoderic acid targeting multiple receptors in cancer: in silico and in vitro study, Tumor Biology, 37 (2016) 14271–14290.
27. M.F. Ahmad, Ganoderma lucidum: A rational pharmacological approach to surmount cancer, Journal of ethnopharmacology, 260 (2020) 113047.
28. S. Gündoğdu, N. Özenver, Anticancer potential of Ganoderma lucidum and its underlying mechanisms, Mushrooms with Therapeutic Potentials: Recent Advances in Research and Development, Springer2023, pp. 221–240.
29. N.-Y. Lee, W.-C. Ko, P.-R. Hsueh, Nanoparticles in the treatment of infections caused by multidrug-resistant organisms, Frontiers in pharmacology, 10 (2019) 1153.
30. I. Babushkina, E. Gladkova, S. Belova, I. Norkin, Application of preparations containing copper nanoparticles for the treatment of experimental septic wounds, Bulletin of Experimental Biology and Medicine, 164 (2017) 162–164.
31. Ryu, Da Hye, Jwa Yeong Cho, Muhammad Hamayun, Dong Gyu Lee, and Ho-Youn Kim. "Metabolic modulation of basil (Ocimum basilicum L.): An insight into growth, metabolomics and antioxidant activity under varying temperature and light conditions." Chemical and Biological Technologies in Agriculture 11, no. 1 (2024): 10.
32. P. Kumari, H. Kumar, J. Kumar, M. Sohail, K. Singh, K. Prasad, Biosynthesized Zinc Oxide nanoparticles control the growth of Aspergillus flavus and its aflatoxin production, International Journal of Nano Dimension, 10 (2019) 320–329.
33. Moradkhah, Shekoufeh, Mohaddeseh Larypoor, and Abdollah Allahverdi. "Design and Fabrication of a Rapid Detection Biosensor for Candida albicans Diagnosis in a Microfluidic Platform." Talanta Open (2025): 100478.
34. Atefi, Aref, et al. "Synthesis, Characterization and Biological Evaluation of a Sulfonamide-Based Antimicrobial Agent." Chemistry Africa (2025): 1-13.
35. D. Devipriya, S.M. Roopan, Cissus quadrangularis mediated ecofriendly synthesis of copper oxide nanoparticles and its antifungal studies against Aspergillus niger, Aspergillus flavus, Materials Science and Engineering: C, 80 (2017) 38–44.
36. S. Maqsood, S. Qadir, A. Hussain, A. Asghar, R. Saleem, S. Zaheer, N. Nayyar, Antifungal properties of copper nanoparticles against Aspergillus niger, Sch. Int. J. Biochem, 3 (2020) 87–91.
37. Das, Prince Edwin, Imad A. Abu-Yousef, Amin F. Majdalawieh, Srinivasan Narasimhan, and Palmiro Poltronieri. "Green synthesis of encapsulated copper nanoparticles using a hydroalcoholic extract of Moringa oleifera leaves and assessment of their antioxidant and antimicrobial activities." Molecules 25, no. 3 (2020): 555.
38. Larypoor, Mohaddeseh, et al. The synergistic effects of chitosan and Ganoderma Lucidum nanoparticles on p53 and HER3 gene expression, International Journal of Molecular and Clinical Microbiology 14.2 (2024): 2061-2071.
39. Prathivadi Bayankaram, Padmaja, and Periyar selvam Sellamuthu. "Antifungal and anti-aflatoxigenic effect of probiotics against Aspergillus flavus and Aspergillus parasiticus." Toxin Reviews 35, no. 1-2 (2016): 10-15.