Food- borne fungi cause serious spoilage and some species can produce toxic metabolites (mycotxins), produce serious health problems. Fungi were isolated from forty samples of fresh Karish cheese, collected from Giza governorate, using malt extract and potato dextrose agar media at 25 ^ΟC. The results indicate that members of Penicillum spp. and  Aspergillus spp. were the mainly contaminated molds of all the tested samples. Members of Penicillum spp. (P. roqueforti, P.corylophilum and P. chrysogenum) and Aspergillus spp. (A. flavus and A. niger). Spectrum of antifungal activity of Lactobacillus casei CH-1, L. bulgaricus CH-2 and L. plantarum (ATCC 8014) were investigated. Among tested strains L.plantarum (ATCC 8014) strain had a strongest inhibitory activity against all isolated of molds from Karish cheese samples. The disc diffusion method was used to evaluate the zone of fungal growth inhibition at various volume of cell-free supernatant. Minimal inhibition concentration (MIC) and minimal fungicidal concentration (MFC) of cell-free supernatant were determined. Cell-free supernatant of L.plantarum (ATCC 8014) culture in Karish cheese was studied. No moldes could be observed till 30 days of storage. Therefore, addition of cell-free supernatant from L.plantarum  (ATCC 8014) to Karish cheese is a good bio-preservatives, preventing fungal spoilage and consequently mycotoxin formation in Karish cheese and recommended to extent the shelf life.