Sugarcane juice is considered the most popular fresh juice in Egypt, with cane juice shops spreading through all the Egyptian cities. Sugarcane juice contains 75 - 85% water and 10 - 21% sucrose. Sugarcane is a suitable host for many saprophytic fungi. No or little information has been reported on fungal flora and their toxins associated sugarcane juice in Egypt so, This study aimed to investigate the natural occurrence of toxigenic fungi associated sugarcane juice. Randomized of seven sugarcane juice samples were collected from seven different localities (Places) in Kalubya Governorate (Banha, Tukh and Shibin El Quanater ). Fungal flora were isolated by serial dilution technique. The obtained data resulted that, isolation from seven different sugarcane juice localities yielded 322 fungal isolates. Higher total fungal count was recorded with location 6 sample. Identification indicated that, five fungal genera belonging to eight species were identified. These are Alternaria alternata, Aspergillus spp., (Aspergillus niger, A. flavus & A. parasiticus), Fusarium spp. (Fusarium solani & F. oxysporum), Penicillium sp. and Rhizopus stolonifer. Aspergillus species were the highest frequently present in sugarcane juice and Aspergillus niger was higher fungal frequency occurred. According to HPLC data, tested of mycotoxins production presented, Four isolates of Aspergillus parasiticus and two isolates of A. flavus were aflatoxins (Afs) producers. Higher aflatoxin quantity (2.91ng/mL) was produced by A. parasiticus (isolate No. 20) from location two of sugarcane juice samples. Whereas, four isolates of Aspergillus niger isolates were positive producer of Ochratoxin A (OTA). Higher Ochratoxin A (OTA) production was recorded with A. niger, isolate No. 25 isolated from location one which gave 2.03 ng/ml of Ochratoxin A (OTA). All Fusarium spp. isolates were negative producer any toxins.