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The cultivation of shiitake mushrooms (Lentinula edodes) is increasing rapidly due to their nutritional and medicinal benefits. Previously, we evaluated different substrates for cultivating L. edodes mushrooms but did not determine the impact of these substrates on the nutritional contents. Thus, the aim of this study was to evaluate the nutritional values of Lentinula edodes mushrooms cultivated on seven different substrates. Samples of fruiting bodies were oven-dried at 45°C for 72 h and grinded by a mill to a particle size of <1 mm. The milled fruiting bodies were analyzed following a well established methods. Accordingly, the nutritional composition and mineral elements of mushrooms cultivated on each of the substrate were determined using a Plasma Spectrophotometer and Microwave Plasma Atomic Emission Spectroscopy, respectively. The content of crude protein, crude fiber, crude fat, and carbohydrates were in the range of 13.67–19.60 %, 12.99–15.34 %, 0.77–1.83, and 52.17–59.39 %, respectively. The highest value of nutrients was obtained from sugarcane bagasse supplemented with poultry manures (S4), except carbohydrates from sugarcane bagasse alone (S1). Therefore, the use of plant substrates supplemented with animal origin improved the nutritional values of L. edodes mushroom in order to combat malnutrition and contribute to food security in the country.