Thirty sugarcane bagasse samples were collected from different sugarcane juice mills in Sohag Governorate, Upper Egypt. The moisture content of tested samples was fluctuated between 32.58 - 70.59%. Total carbohydrates were estimated, and the richest sample was No. 21 with 535.10 mg/g dry weight, whereas the lowest sample was No. 3 with 508.131 mg/g dry weight. Soluble sugars of different samples were determined using spectrophotometer at 620 nm. The soluble sugars content ranged between 120 - 160 mg/g dry weight. Dilution- (8 genera, 27 species and 4 varieties) and direct- (8 genera, 25 species and 4 varieties) plate methods on glucose-Czapek's agar medium at 28 ±2 oC were used for isolation of bagasse glucophilic fungi. It was found that the total fungal populations in case of dilution–plate method (69720 cfu/g) was so high than those of direct–plate method (828 cfu/ 20 bagasse segments). Aspergillus was appeared in high population and was parallel to those of total fungal counts (51700 cfu/g, 74.15 % of total count, in 29 samples with 96.7% of the samples in dilution method and 731 cfu/ 20 segments, 88.3% of total count, in 30 samples with 100% of the tested samples using direct-plate method.