40 milk samples were collected from cattle suffering from chronic mastitis with no or poor response to antibiotic therapy with common antibiotics, these samples were subjected tobacteriological and mycological culture then confirmed by traditional fungal confirmatory procedures and multiplex PCR (m-PCR). The resultsshowed 37 positive samples for fungal culture hence C. albicans took the lead of them by 60 % , A. fumigatus 22.5% , A. niger 10 % while C. neoformans not detected in any sample. Mixed bacterial and fungal infectionwas detected in 83.78% whilefungi as a sole etiological agent were detected in 16.22% of samples. m PCR succeeded to distinguish between A. fumigatus , A. niger and C. albicans more accurate and fast than traditional confirmatory methods .Pure mycotic mastitis cases were subjected to treatment by a new novel drug consisting of extract of medicinal plants (garlic and sesame oil). All treated cases responded well to this drug and returned to normal udder condition and to full milk production.
Key words: PCR, candida,Asperigillus, Mycotic Mastitis.
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