The potential of using ensiling, with and without supplemental enzymes, as a cost-effective pretreatment for bioethanol production from agricultural residues was investigated. Ensiling did not significantly affect the lignin content of barley straw, cotton stalk, and triticale hay ensiled without enzyme, but slightly increased the lignin content in triticale straw, wheat straw, and triticale hay ensiled with enzyme. The holocellulose (cellulose plus hemicellulose) losses in the feedstocks, as a result of ensiling, ranged from 1.31 to 9.93%. The percent holocellulose loss in hays during ensiling was lower than in straws and stalks. Ensiling of barley, triticale, wheat straws, and cotton stalk significantly increased the conversion of holocellulose to sugars during subsequent hydrolysis with two enzyme combinations. Enzymatic hydrolysis of ensiled and untreated feedstocks by Celluclast 1.5 L-Novozyme 188 enzyme combination resulted in equal or higher saccharification than with Spezyme CP-xylanase combination. Enzyme loadings of 40 and 60 FPU/g reducing sugars gave similar sugar yields. The percent saccharification with Celluclast 1.5 L-Novozyme 188 at 40 FPU/g reducing sugars was 17.1 to 43.6%, 22.4 to 46.9%, and 23.2 to 32.2% for untreated feedstocks, feedstocks ensiled with, and without enzymes, respectively. Fermentation of the hydrolysates from ensiled feedstocks resulted in ethanol yields ranging from 0.21 to 0.28 g/g reducing sugars.