Investigation of the Antimicrobial and Antioxidant Activities of the Ethanol Extract of Amanita muscaria

Öz

Characterized by its bright red cap adorned with white spots, Amanita muscaria is not only visually distinctive but also of considerable pharmaceutical interest due to its diverse array of biologically active compounds. Rich in metabolites such as muscimol, ibotenic acid, phenolic compounds, and polysaccharides, this species has been regarded as a potential source of antioxidant and antimicrobial agents. In light of the growing global challenge of antibiotic resistance, the exploration of natural sources has gained increasing significance, with fungal species emerging as promising alternatives. Different concentrations of the A. muscaria extract were tested against 12 reference, 11 multidrug-resistant (MDR), 13 clinical, and 6 food-isolated microorganisms. Disk diffusion assays revealed that the extract produced inhibition zones ranging from 7 to 10 mm, particularly against Listeria monocytogenes, Pseudomonas fluorescens, Enterococcus faecium, and Staphylococcus aureus. The minimum inhibitory concentration (MIC) values ranged between 310 and 5,080 µg/mL, while the minimum bactericidal concentrations (MBCs) were generally above 10,000 µg/mL, indicating a predominantly bacteriostatic effect. The relatively weak activity observed against Gram-negative bacteria was attributed to the diffusion-limiting properties of their lipopolysaccharide outer layer. Antioxidant capacity was assessed using the DPPH radical scavenging assay, in which the extract was compared with ascorbic acid. At 1000 µg/mL, A. muscaria exhibited 97.05% inhibition, comparable to that of ascorbic acid (94.67%), and demonstrated stronger radical scavenging potential at lower concentrations (EC₅₀ = 33.5 µg/mL). These findings suggest that while the antimicrobial potential of A. muscaria is limited, its antioxidant activity is remarkably high. The results highlight the potential of this species as a valuable natural source of antioxidant compounds and indicate that its antimicrobial efficacy may be enhanced through the isolation of active constituents, combination strategies, or nanoparticle-based approaches.

Anahtar Kelimeler

• Amanita muscaria
• Mushroom
• Antimicrobial activity
• Antioxidant activity
• Microorganism

Amanita muscaria etanol ekstraktının antimikrobiyal ve antioksidan aktivitelerinin incelenmesi

Öz

Kırmızı şapkası ve beyaz benekleriyle doğada kolayca tanınan Amanita muscaria, yalnızca görsel olarak değil, içerdiği biyolojik olarak aktif bileşikler sayesinde farmasötik açıdan da dikkat çeken bir mantar türüdür. Muscimol, ibotenik asit, fenolik bileşikler ve polisakkaritler gibi metabolitler bakımından zengin olan bu tür, potansiyel antioksidan ve antimikrobiyal ajan kaynağı olarak değerlendirilmektedir. Günümüzde artan antibiyotik direnci sorununa karşı doğal kaynakların araştırılması önem kazanmış; mantar türleri bu kapsamda öne çıkan alternatifler arasında yer almıştır. Etanol ile elde edilen A. muscaria ekstraktının farklı konsantrasyonları 12 referans, 11 çoklu ilaç dirençli (MDR), 13 klinik ve 6 gıda kaynaklı mikroorganizmaya karşı çalışılmıştır. Disk difüzyon analizleri, ekstraktın özellikle Listeria monocytogenes, Pseudomonas fluorescens, Enterococcus faecium ve Staphylococcus aureus suşlarında 7–10 mm aralığında inhibisyon zonları oluşturduğunu göstermiştir. Minimum inhibitör konsantrasyon değerlerinin 310–5.080 µg/mL aralığında, minimum bakterisidal konsantrasyonların ise genellikle 10.000 µg/mL’nin üzerinde bulunması, ekstraktın bakteriyostatik etkiye sahip olduğunu ortaya koymuştur. Gram-negatif bakterilerde etkinliğin zayıf olması, lipopolisakkarit tabakasının difüzyon engelleyici özelliğiyle ilişkilendirilmiştir. Antioksidan kapasite belirlemesinde, ekstraktın DPPH radikal giderme aktivitesi askorbik asit ile kıyaslanmıştır. 1000 µg/mL konsantrasyonda A. muscaria %97,05 inhibisyon ile askorbik aside (%94,67) eşdeğer düzeyde etki göstermiş; düşük konsantrasyonlarda ise daha güçlü radikal süpürme potansiyeli sergilemiştir (EC₅₀ = 33,5 µg/mL). Elde edilen sonuçlar, A. muscaria’nın antimikrobiyal yönünün sınırlı, ancak antioksidan etkinliğinin oldukça yüksek olduğunu göstermektedir. Elde edilen bulgular, bu mantarın doğal antioksidan bileşiklerin önemli bir kaynağı olabileceğini, ayrıca antimikrobiyal etkinliğinin saf bileşiklerin ayrıştırılması, birlikte kullanım stratejileri veya nanoparçacık temelli yöntemlerle geliştirilebileceğini göstermektedir.

Anahtar Kelimeler

• Amanita muscaria
• Mantar
• Antimikrobiyal aktivite
• Antioksidan aktivite
• Mikroorganizma

Kaynakça

1. Andrews, J. M. (2001). Determination of minimum inhibitory concentrations. Journal of Antimicrobial Chemotherapy, 48(Suppl. 1), 5–16. https://doi.org/10.1093/jac/48.suppl_1.5
2. Archana, O., & Nagadesi, P. K. (2024). Mushroom alkaloids as nutraceuticals, bioactive and medicinal properties: A preliminary review. Plant Science Today, 11(4), 651–661. https://doi.org/10.14719/pst.3161
3. Benek A. (2024) İzmir Bozdağlar Silsilesinin Karayosunu Florasının Belirlenmesi ve Bazı Örneklerin Biyolojik Aktivitelerinin İncelenmesi(Doctoral Thesis)
4. Benek, A., Şenturan, M., Şimşek, Ö., … Canlı, K. (2021). Bazı Karayosunu Türlerinin İn Vitro Antimikrobiyal Aktivitelerinin Belirlenmesi. Duzce University Journal of Science and Technology, 9(3), 474-481. https://doi.org/10.29130/dubited.872689
5. Bentharavithana, J., Islam, T., & Xu, B. (2025). Medicinal mushrooms in colon cancer therapy: Mechanisms of action of bioactive compounds and therapeutic potential. International Journal of Molecular Sciences, 26(11), 5304. https://doi.org/10.3390/ijms26115304
6. Bhambri, A., Srivastava, M., Mahale, V. G., Mahale, S., & Karn, S. K. (2022). Mushrooms as potential sources of active metabolites and medicines. Frontiers in Microbiology, 13, 837266. https://doi.org/10.3389/fmicb.2022.837266
7. Birben, E., Sahiner, U. M., Sackesen, C., Erzurum, S., & Kalayci, O. (2012). Oxidative stress and antioxidant defense. World Allergy Organization Journal, 5(1), 9–19. https://doi.org/10.1097/WOX.0b013e3182439613
8. Borlaza, C. B. (2019, July). Antibacterial potential of fly agaric (Amanita muscaria) synthesized with silver nanoparticles (AgNP) against Salmonella typhimurium. Paper presented at the 2nd International Conference on Multidisciplinary Academic Research (ICMAR-2019), Bali, Indonesia.

9. Canli, K.; Turu, D.; Benek, A.; Bozyel, M.E.; Simsek, Ö.; Altuner, E.M. Biochemical and Antioxidant Properties as well as Antimicrobial and Antibiofilm Activities of Allium scorodoprasum subsp. jajlae (Vved.) Stearn. Curr. Issues Mol. Biol. 2023, 45, 4970–4984. https://doi.org/10.3390/cimb45060316
10. Carboué, Q., & Lopez, M. (2021). Amanita muscaria: Ecology, chemistry, myths. Encyclopedia, 1(3), 905–914. https://doi.org/10.3390/encyclopedia1030069
11. Chelela, B. L., Chacha, M., & Matemu, A. (2014). Wild edible mushroom value chain for improved livelihoods in Southern Highlands of Tanzania. American Journal of Research Communication, 2(8), 1–14. http://www.usa-journals.com/wp-content/uploads/2014/07/Chelela_Vol28-2.pdf
12. Chun, S., Gopal, J., & Muthu, M. (2021). Antioxidant Activity of Mushroom Extracts/Polysaccharides-Their Antiviral Properties and Plausible AntiCOVID-19 Properties. Antioxidants (Basel, Switzerland), 10(12), 1899. https://doi.org/10.3390/antiox10121899
13. Delcour, A. H. (2009). Outer membrane permeability and antibiotic resistance. Biochimica et Biophysica Acta (BBA) – Proteins and Proteomics, 1794(5), 808–816. https://doi.org/10.1016/j.bbapap.2008.11.005
14. Eda Tapan, Süfer, Ö., Taşkin, H. et al. Antioxidant Activities of Some Edible and Poisonous Amanita Species from Türkiye. Biol Bull Russ Acad Sci 51, 1662–1670 (2024). https://doi.org/10.1134/S1062359023603105
15. Ersin, M., Gönül, E., Duman, E., Gül, G., Turu, D., Benek, A., & Canlı, K. (2025). Investigation of Plantago lanceolata L.: A multidimensional study on its biochemical profiling, antioxidant capacity, and biological activities. Communications Faculty of Sciences University of Ankara Series C: Biology, 34(1), 76–103. https://doi.org/10.34074/commfacscibio.1654646
16. Falandysz, J., & Treu, R. (2019). Amanita muscaria: Bio-concentration and bio-indicative potential for metallic elements. Environmental Earth Sciences, 78, 722. https://doi.org/10.1007/s12665-019-8718-x
17. Ferreira, I. C. F. R., Barros, L., & Abreu, R. M. V. (2009). Antioxidants in wild mushrooms. Current Medicinal Chemistry, 16(12), 1543–1560. https://doi.org/10.2174/092986709787909587
18. Grochowska, J., Grygoruk, N., & Weiner, M. (2025). Harmfulness and potential health benefits of the fly agaric (Amanita muscaria): A literature review. Health Problems of Civilization, 19(1), 95–106. https://doi.org/10.5114/hpc.2025.146651
19. Kalač P. (2013). A review of chemical composition and nutritional value of wild-growing and cultivated mushrooms. Journal of the science of food and agriculture, 93(2), 209–218. https://doi.org/10.1002/jsfa.5960
20. Kosanić, M., Ranković, B., & Dašić, M. (2012). Mushrooms as possible antioxidant and antimicrobial agents. Iranian Journal of Pharmaceutical Research: IJPR, 11(4), 1095–1102. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3813180/
21. Liguori, I., Russo, G., Curcio, F., Bulli, G., Aran, L., Della-Morte, D., Gargiulo, G., Testa, G., Cacciatore, F., Bonaduce, D., & Abete, P. (2018). Oxidative stress, aging, and diseases. Clinical interventions in aging, 13, 757–772. https://doi.org/10.2147/CIA.S158513
22. Liuzzi, G. M., Petraglia, T., Latronico, T., Crescenzi, A., & Rossano, R. (2023). Antioxidant compounds from edible mushrooms as potential candidates for treating age-related neurodegenerative diseases. Nutrients, 15(8), 1913. https://doi.org/10.3390/nu15081913
23. Martins, T., Machado-Carvalho, L., Aires, A., Saavedra, M. J., & Marques, G. (2023). Antioxidant, Antimicrobial and Cosmeceutical Potential of Wild Mushroom Extracts. Applied Microbiology, 3(2), 562-579. https://doi.org/10.3390/applmicrobiol3020040
24. Mensor, L. L., Menezes, F. S., Leitão, G. G., Reis, A. S., dos Santos, T. C., Coube, C. S., & Leitão, S. G. (2001). Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method. Phytotherapy Research, 15(2), 127–130. https://doi.org/10.1002/ptr.687
25. Michelot, D., & Melendez-Howell, L. M. (2003). Amanita muscaria: chemistry, biology, toxicology, and ethnomycology. Mycological research, 107(Pt 2), 131–146. https://doi.org/10.1017/s0953756203007305
26. Mookherjee, N., Anderson, M. A., Haagsman, H. P., & Davidson, D. J. (2023). Antimicrobial host defence peptides: Functions and clinical potential. Nature Reviews Drug Discovery, 22(2), 87–108. https://doi.org/10.1038/s41573-019-0058-8
27. Munteanu, I. G., & Apetrei, C. (2021). Analytical Methods Used in Determining Antioxidant Activity: A Review. International Journal of Molecular Sciences, 22(7), 3380. https://doi.org/10.3390/ijms22073380
28. Nikaido, H. (2003). Molecular basis of bacterial outer membrane permeability revisited. Microbiology and Molecular Biology Reviews, 67(4), 593–656. https://doi.org/10.1128/MMBR.67.4.593-656.2003
29. Pięta-Chrystofiak, M., & Brohs, D. (2023). Red fly agaric (Amanita muscaria) consumption among members of Internet discussion groups. Studia Paedagogica Ignatiana, 26(2), 195–221.
30. Rampolli, F. I., Kamler, P., Carnevale Carlino, C., & Bedussi, F. (2021). The deceptive mushroom: Accidental Amanita muscaria poisoning. European Journal of Case Reports in Internal Medicine, 8(3), 002212. https://doi.org/10.12890/2021_002212
31. Satora, L., Pach, D., Butryn, B., Hydzik, P., & Balicka-Ślusarczyk, B. (2005). Fly agaric (Amanita muscaria) poisoning: Case report and review. Toxicon, 45(7), 941–943. https://doi.org/10.1016/j.toxicon.2005.01.005
32. Silhavy, T. J., Kahne, D., & Walker, S. (2010). The bacterial cell envelope. Cold Spring Harbor perspectives in biology, 2(5), a000414. https://doi.org/10.1101/cshperspect.a000414
33. Singh, A., Saini, R. K., Kumar, A., Chawla, P., & Kaushik, R. (2024). Mushrooms as nutritional powerhouses: A review of their bioactive compounds, health benefits, and value-added products. Foods, 14(5), 741. https://doi.org/10.3390/foods14050741
34. Stoeva-Grigorova, S., Yarabanova, I., Radeva-Ilieva, M., Ivanova, D., Zlateva, S., & Marinov, P. (2025). Acute Amanita muscaria Toxicity: A Literature Review and Two Case Reports in Elderly Spouses Following Home Preparation. Toxins, 17(12), 570. https://doi.org/10.3390/toxins17120570
35. Tazehkand, M. N., & Yılmaz, M. B. (2019). Investigation of antibacterial effects of chlorzoxazone. Çukurova Medical Journal, 44(2), 653–658. https://doi.org/10.17826/cumj.459036
36. Tunca-Pınarlı, Y., Benek, A., Turu, D., Bozyel, M. E., Canlı, K., & Altuner, E. M. (2023). Biological activities and biochemical composition of endemic Achillea fraasii. Microorganisms, 11(4), 978. https://doi.org/10.3390/microorganisms11040978
37. Tunç, K., Semerci, A. B., & Okur, İ. (2020). Antioxidant activity of the fruits of Pyracantha coccinea using ethanolic extract method. Food and Health, 6(1), 35-40. https://doi.org/10.3153/FH20005
38. Van Boeckel, T. P., Brower, C., Gilbert, M., Grenfell, B. T., Levin, S. A., Robinson, T. P., Teillant, A., & Laxminarayan, R. (2015). Global trends in antimicrobial use in food animals. Proceedings of the National Academy of Sciences of the United States of America, 112(18), 5649–5654. https://doi.org/10.1073/pnas.1503141112
39. Voynova M, Shkondrov A, Krasteva I, Kondeva-Burdina M (2021) In vitro effects of synthetic muscimol and an extract from Amanita muscariaon human recombinant MAOB enzyme. Pharmacia 68(1): 147-150. https://doi.org/10.3897/pharmacia.68.e60705
40. Wagner, A., Pehar, M., Yan, Z., & Kulka, M. (2023). Amanita muscariaextract potentiates production of proinflammatory cytokines by dsRNA-activated human microglia. Frontiers in pharmacology, 14, 1102465. https://doi.org/10.3389/fphar.2023.1102465
41. World Health Organization. (2017). WHO publishes list of bacteria for which new antibiotics are urgently needed.Geneva: WHO.
42. World Health Organization. (2022). Global burden of bacterial antimicrobial resistance in 2019: A systematic analysis.Geneva: WHO.
43. World Health Organization. (2024). Global Antimicrobial Resistance and Use Surveillance System (GLASS) report 2024.Geneva: WHO.
44. Yang, J. Y., Tao, L., Lou, D., Patabendige, N. M., Ediriweera, A. N., Liu, S., Lu, W., Tarafder, E., Rapior, S., & Hapuarachchi, K. K. (2025). Innovative applications of medicinal mushrooms in functional foods and nutraceuticals: a focus on health-boosting beverages. Frontiers in cellular and infection microbiology, 15, 1605301. https://doi.org/10.3389/fcimb.2025.1605301
45. Zhang, P., Tang, L. P., Cai, Q., & Xu, J. P. (2015). A review on the diversity, phylogeography and population genetics of Amanita mushrooms. Mycology, 6(2), 86–93. https://doi.org/10.1080/21501203.2015.1042536
46. Zhen, X., Stålsby Lundborg, C., Sun, X., Zhu, N., Gu, S., & Dong, H. (2021). Economic burden of antibiotic resistance in China: a national level estimate for inpatients. Antimicrobial resistance and infection control, 10(1), 5. https://doi.org/10.1186/s13756-020-00872-w