TOXICOLOGICAL EVALUATION OF LEPISTA NUDA (BULL. EX FR.) COOKE MYCELIUM PRODUCED BY AN IN VITRO CULTURE METHODOLOGY
Objective: Considering the interest in L. nuda as a source of ingredients for the development of functional food and nutraceuticals has increased, the objective of this study was to evaluate its general toxicity and possible genotoxic effects in rats to assess its safety.
Methods: This study evaluated the safety of L. nuda mycelium by using genotoxicity assays (reverse mutation, chromosomal aberration, and micronuclei tests) and a short-term toxicity test.
Results: Our results have indicated that L. nuda mycelium did not significantly increase the number of revertant colonies and chromosomal aberration in both in vitro assays. Furthermore, it did not induce any increase in micronuclei formation in mouse bone marrow.
Conclusion: In summary, no mutagenic effects and no evidence of systemic toxicity were found in this safety assessment, and the use of L. nuda mycelia is safe at a dose of 3 g/kg body weight in S-D rats. Using a safety factor of 100, the calculated acceptable daily intake in humans is 30 mg/kg body weight/d.
2. Lindequist U, Niedermeyer TH, Julich WD. The pharmacological potential of mushrooms. J Evidence-Based Complementary Altern Med 2005;2:285-99.
3. Sapozhnikova Y, Byrdwell WC, Lobato A, Romig B. Effects of UV-B radiation levels on concentrations of phytosterols, ergothioneine, and polyphenolic compounds in mushroom powders used as dietary supplements. J Agric Food Chem 2014;62:3034-42.
4. Borchers AT, Keen CL, Gershwin ME. Mushrooms, tumors, and immunity: an update. Exp Biol Med (Maywood) 2004;229:393-406.
5. Barros L, Venturini BA, Baptista P, Estevinho LM, Ferreira IC. Chemical composition and biological properties of portuguese wild mushrooms: a comprehensive study. J Agric Food Chem 2008;56:3856-62.
6. NoÃ«l-Suberville C, Cruz C, Guinberteau J, Montury M. Correlation between fatty acid content and aromatic compound release in fresh blewit (Lepista nuda). J Agric Food Chem 1996;44:1180-3.
7. Beattie KD, Ulrich R, Grice ID, Uddin SJ, Blake TB, Wood KA, et al. Ethanolic and aqueous extracts derived from Australian fungi inhibit cancer cell growth in vitro. Mycologia 2011;103:458-65.
8. Chen MH, Li WS, Lue YS, Chu CL, Pan IH, Ko CH, et al. Clitocybe nuda activates dendritic cells and acts as a DNA vaccine adjuvant. J Evidence-Based Complementary Altern Med 2013; doi:10.1155/2013/761454. [Article in Press]
9. Mercan N, Duru M, Turkoglu A, Gezer K, Kivrak I, Turkoglu H. Antioxidant and antimicrobial properties of ethanolic extract fromLepista nuda (Bull.) Cooke. Ann Microbiol 2006;56:339-44.
10. Dulger B, Ergul CC, Gucin F. Antimicrobial activity of the macrofungus lepista nuda. Fitoterapia 2002;73:695-7.
11. Chen MH, Lin CH, Shih CC. Antidiabetic and antihyperlipidemic effects of clitocybe nuda on glucose transporter 4 and AMP-activated protein kinase phosphorylation in High-Fat-Fed mice. J Evidence-Based Complementary Altern Med 2014; doi:10.1155/2014/981046. [Article in Press]
12. Yaoita Y, Matsuki K, Iijima T, Nakano S, Kakuda R, Machida K, et al. New sterols and triterpenoids from four edible mushrooms. Chem Pharm Bull (Tokyo) 2001;49:589-94.
13. Yaoita Y, Kohata R, Kakuda R, Machida K, Kikuchi M. Ceramide constituents from five mushrooms. Chem Pharm Bull (Tokyo) 2002;50:681-4.
14. Lee WY, Park EJ, Ahn JK, Ka KH. Ergothioneine contents in fruiting bodies and their enhancement in mycelial cultures by the addition of methionine. Mycobiology 2009;37:43-7.
15. Mukhtar H, Ijaz SS, Ikram ul H. Upstream and downstream processing of lovastatin by aspergillus terreus. Cell Biochem Biophys 2014;70:309-20.
16. Nielsen J, Johansen CL, Jacobsen M, Krabben P, Villadsen J. Pellet formation and fragmentation in submerged cultures of Penicillium chrysogenum and its relation to penicillin production. Biotechnol Prog 1995;11:93-8.
17. Max B, Salgado JM, Rodriguez N, Cortes S, Converti A, Dominguez JM. Biotechnological production of citric acid. Braz J Microbiol 2010;41:862-75.
18. International A: Official methods of analysis of AOAC International. Official methods of analysis of AOAC International. [Gaithersburg, Md.]: AOAC International; 1995.
19. OECD: Test No. 471: Bacterial Reverse Mutation Test. Paris, France: Organization for Economic Cooperation and Development; 1997.
20. Maron DM, Ames BN. Revised methods for the Salmonella mutagenicity test. Mutat Res 1983;113:173-215.
21. OECD: Test No. 473: In vitro Mammalian Chromosome Aberration Test. Paris, France: Organization for Economic Cooperation and Development; 1997.
22. OECD: Test No. 474: Mammalian Erythrocyte Micronucleus Test. OECD Publishing; 1997.
23. OECD: Test No. 407: Repeated Dose 28-day Oral Toxicity Study in Rodents. Paris, France: Organization for Economic Cooperation and Development; 2001.
24. Lee CY, Park JE, Kim BB, Kim SM, Ro HS. Determination of mineral components in the cultivation substrates of edible mushrooms and their uptake into fruiting bodies. Mycobiology 2009;37:109-13.
25. Papaspyridi LM, Aligiannis N, Topakas E, Christakopoulos P, Skaltsounis AL, Fokialakis N. Submerged fermentation of the edible mushroom Pleurotus ostreatus in a batch stirred tank bioreactor as a promising alternative for the effective production of bioactive metabolites. Molecules 2012;17:2714-24.
26. Chang ST, Lau OW, Cho KY. The cultivation and nutritional value of Pleurotus sajor-caju. European journal of applied microbiology and biotechnology 1981;12:58-62.
27. Preeti A, Pushpa S, Sakshi S, Jyoti A. Antioxidant mushrooms: a review. Int Res J Pharm 2012;3:65-70.
28. Grundemann D. The ergothioneine transporter controls and indicates ergothioneine activity--a review. Prev Med 2012;54 Suppl:S71-4.
29. Ulziijargal E, Yang JH, Lin LY, Chen CP, Mau JL. Quality of bread supplemented with mushroom mycelia. Food Chem 2013;138:70-6.
30. Kalac P, Niznanska M, Bevilaqua D, Staskova I. Concentrations of mercury, copper, cadmium and lead in fruiting bodies of edible mushrooms in the vicinity of a mercury smelter and a copper smelter. Sci Total Environ 1996;177:251-8.
31. Mathijs K, Brauers KJ, Jennen DG, Lizarraga D, Kleinjans JC, van Delft JH. Gene expression profiling in primary mouse hepatocytes discriminates true from false-positive genotoxic compounds. Mutagenesis 2010;25:561-8.
32. Mortelmans K, Zeiger E. The Ames Salmonella/microsome mutagenicity assay. Mutat Res 2000;455:29-60.
33. Zbinden G, Flury-Roversi M. Significance of the LD50-test for the toxicological evaluation of chemical substances. Arch Toxicol 1981;47:77-99.
34. Petterino C, Argentino-Storino A. Clinical chemistry and haematology historical data in control Sprague-Dawley rats from pre-clinical toxicity studies. Exp Toxicol Pathol 2006;57:213-9.
35. Lee JM, Lee MA, Do HN, Song YI, Bae RJ, Lee HY, et al. Historical control data from 13-week repeated toxicity studies in Crj: CD (SD) rats. Lab Anim Res 2012;28:115-21.
36. Han ZZ, Xu HD, Kim KH, Ahn TH, Bae JS, Lee JY, et al. Reference data of the main physiological parameters in control sprague-dawley rats from pre-clinical toxicity studies. Lab Anim Res 2010;26:153-64.