Mastitis is an economic food production problem in dairy animals. Bacterial etiology of mastitis is common but mycotic etiology of mastitis is scarce and obscured. The present study aims to use traditional and molecular methods to identify the fungal species that were separated from cattle mastitis, evaluate their susceptibility to antifungals, and investigate the variation between fungi isolated from dairy cows with clinical and subclinical mastitis. Three hundred milk samples were collected from Dakahlia (175) and Kafer Elsheikh (125) governorates. All samples were cultivated on the surface of Malt extract agar, Rice agar medium, Chromogenic agar and Sabouraud's dextrose agar. Fungi were identified using phenotypic characteristics. Aspergillus and Candida species were the most isolated fungi. Positive samples for mycotic growth were (103; 59%) from Dakahlia and (55; 44%) from Kafer El sheikh. In Dakahlia , Aspergillus species was the most common type of isolated mold. Species from clinical (36.3%) and subclinical (22.7%) mastitis while in Kafer Elsheikh were Penicillium (47.5%) and Aspergillus (40 %). The most isolated mold species from clinical mastitis was A. flavus (26%). The isolated yeast from Dakhliea from the clinical milk samples was C. albicans (24%) while in Kafer Elsheikh was C. galabrata (28.6%) . PCR results were C. albicans, Cryptococcus neoformans, A. flavus, A. niger and. A. fumigatus. . It is recommended that an animal unresponsive to antimycotic druggy must be suspected of mycotic mastitis, detailed mycological investigation should be conducted to establish the role of fungi in mastitis.