Abstract: The fungi play an important role in cheese manufacture such as Penicillium roqueforti nevertheless subsequent development of filamentous fungi occurs during cheese ripening on its surface, this accompanied with production of some bad flavors and tastes, as well as some mycotoxins. In this study, 125 samples of cheeses were collected to determination of fungi and aflatoxin B1 (AFB1). The results indicated that percentages of Aspergillus flavus, Aspergillus niger, Aspergillus ocnrecious and Aspergillus oryzae were 18.6, 12.4, 8.6 and 5.3%, respectively. Rhizopus spp., Penicillium spp., Mucor spp., trichoderma spp. and Fusarium moniliforme recorded 22.1, 16.8, 11.5, 2.4 and 0.5%, respectively. Aged Ras cheese was the highest contamination with AFB1 (from 8.3a to 19.5a ppb) average 13.9a ppb, fresh Domiate cheese recorded the lowest contamination with toxin with average 0.95f ppb. A. flavus produced the highest quantity of AFB1 in broth medium for 2 months unlike Penicillium spp. was the lowest Production of toxin (0.6c ppb). Counts of fungi and AFB1 production affected by tested anti-fungi, Ascorbic acid and Broken Saccharomyces cerevisiae Cells after Freezing (BSCAF) were stronger set but Salicylic acid and Citric acid were the weaker set anti-fungi in this point. Aspergillus flavus was the greatest resistant to Salicylic acid.