This survey has been carried out to determine, identify, and isolate fungi by standard blotter method (SBM), especially Aflatoxin-producing fungi; Aspergillus species that produce Aflatoxin B1, B2, G1, and G2 that are secondary metabolites, carcinogenic, and poisonous to humans and animals, from a total of 91 Egyptian spice samples (ginger, nutmeg, red chilli, and paprika) in Dakahlia markets and 11 hospital and medical centers in Mansoura. From all genera estimated, the genus Aspergillus had the highest frequency and prevalence, accounting for 291 of the total number of instances involving the isolation of fungi. Then Pencillium genus ranked second, accounting for 55, and Fusarium genus, accounting for 47. Also, 71 Aspergillus flavus group isolates were screened, and the aflatoxin concentrations of them were quantitatively calculated using HPLC. Moreover, isolates were determined to possess toxigenic properties, with their capacity to manufacture aflatoxins ranging from 15 to 521 ng/ml of culture filtrate. Molecular identification by ITS barcoding of the DNA of the studied strain was submitted to NCBI GenBank. The sequence length for the selected Aspergillus fungus was 457 bp, and the GC content was 58.2%. The inhibitory effect using ten essential oils with different concentrations was as follows: the most effective essential oils were clove oil, cinnamon oil and mint oil  by inhibition ratio at 100% at 0.8% concentration for all of them.