All experimental animals were observed once a day for signs of toxicity, mortality, and morbidity until the completion

of the treatment. The body weight of each animal was recorded at the initiation of the see more treatment and prior to bone marrow sampling. Peripheral blood samples (3 to 4 μL) from tail vein were collected at 48 h and 72 h after dosing and then applied to acridine orange-coated slides for 3 to 4 h at room temperature. The smear samples were microscopically examined with a fluorescent microscope (Zeiss, Oberkochen, Germany) for the number of reticulocytes (orange-red signal), and reticulocytes that contained at least one positive micronucleus (yellow-green signal). The results were expressed as the

frequency of reticulocytes per 1000 red blood cells and micronucleated reticulocytes per 1000 reticulocytes. All values presented throughout this manuscript were expressed as mean ± standard error of mean (SEM). Mean differences between the control and the treatment groups were analyzed by one-way ANOVA followed by Duncan’s test. Aberrant cells from each concentration were compared to the negative control values using Chi-square analyses. EX 527 mw A value of p < 0.05 was considered to be statistically significant. A resurgence of interest in natural products is spreading across many parts of the world currently as they are thought to be new alternative medicines for conventional therapies with fewer side effects. In East Asian countries and more recently in the United States, intensive research has increasingly demonstrated the potential beneficial health properties of compounds extracted from mushrooms for the prevention and management of cancer and other life-debilitating

diseases. For such reasons, the genotoxicity of culinary-medicinal mushrooms that are 4��8C used by the general population should be warranted in order to identify the ingredients that pose mutagenic and carcinogenic risks. To our knowledge, there have been no reports on the mutagenicity of H. erinaceus prior to this paper. Therefore, this is the first report undertaken to evaluate the genotoxicity of a standardized H. erinaceus mycelium enriched with 5 mg/g erinacine A by using the Ames test, the chromosomal aberration test, and the erythrocyte micronucleus test. The Ames test is widely used as an initial screening method to determine the mutagenic potential of newly discovered products since there is a high correlation between the positive responses in test mutagenicity and carcinogenicity [33] and [34]. The proximate analysis and HPLC analysis of EAHE mycelium are shown in Table S1 and Figure S1, respectively. These results are in line with those previously published [27].