Thirty fungal isolates, which include nineteen species and belong to five genera, were isolated from unusual soil samples from Assiut and Wadi El -Natrun, Egypt. All fungal cultures were cultivated on YES medium for secondary metabolites production; the culture filtrates were separately subjected to solvent extraction by: Ethyl Acetate, Petroleum Ether, Dichloromethane and Dichloromethane: Methanol (2: 1); the fungal mat was extracted by Dichloromethane: Methanol (2: 1) only. The obtained Ethyl Acetate extracts of all fungal isolates were tested for their antimicrobial activity against six different pathogenic bacterial strains; three of them were gram positive bacteria (Staphylococcus Aureus, Bacillus Cereus, and Streptococcus Pneumonia); the others were gram negative bacteria (Sertatia Marcescens, E. Coli and Salmonella Typhi). In addition, against five pathogenic fungal strains (Aspergillus Candidus, Aspergillus Flavus, Aspergillus Niger, Aspergillus Terreus and Geotrichum Candidum), the tested Ethyl Acetate fungal extract showed marked antimicrobial activity against most of the tested strains. On the other hand, the highest antibacterial activity was recorded in case of the Penicillium Chrysogenum extract, but the highest antifungal activity was recorded in case of the Penicillium Aurantiogriseum extract. The secondary metabolites of all fungal isolates were separated on TLC plates, and the most four active strains were completely identified as Aspergillus Candidus, Aspergillus Terreus, Penicillium Aurantiogriseum and Penicillium Chrysogenum. Furthermore, the antimicrobial, anti-insect, anti-herbal and anti-tumor activities of its extracts were evaluated.