The objective of this study was to evaluate the production (P) (g of fresh mushrooms /bag) and biological efficiency (BE) (g of fresh mushrooms per 100 g of dry substrate) of 17 fungal strains, namely Pleurotus ostreatus Po-IAP, Po-P38, Po-P14, Po-IE202, Po-Sfco, Po-JP, Po-Psma, and Po-POS, Pleurotus djamour Pd-PRO and Pd-UTMR, Pleurotus eryngii Pe-MB and Pe-PQ, Lentinula edodes L15, L9, L5, and LC, and Hericium erinaceus Heri, produced in corn stover (CS) and to assess the content of crude protein (CP), lignin (L), cellulose, hemicellulose, acid detergent fiber (ADF), and neutral detergent fiber (NDF) and in vitro digestibility of dry matter (IVDMD) of the residual substrate of CS, the so called spent mushroom substrate (SMS), in comparison to the non-inoculated substrate (C). The variables were analyzed as a completely randomized block design using R 4.0.3 software. Means were compared using Tukey’s procedure. The Pleurotus spp. strains, compared to Lentinula spp. and Hericium spp., presented better BE and P. In comparison to C, the SMS increased the CP content (p < 0.05) from 10.8% (Po-JP) to 70.3% (LC), while NDF decreased (p < 0.05) from 11.5% (Pd-Pro) to 33.5% (L15) and IVDMD increased (p < 0.05) from 16.2% (Heri) to 47.7% (Pd-UTMR). In conclusion, of the 17 strains evaluated, the 3 strains of Lentinula edodes (L5, L15, and L9), one strain of Pleurotus djamour (Pd-UTMR), and one strain of Pleurotus ostreatus (Po-IAP) generated a SMS that, due to its nutritional improvement and increase in IVDMD, could be used as feed for ruminants. Our results also showed that corn stover is a suitable substrate to produce Pleurotus spp. fruiting bodies, with strain Po-IAP as the best yielding.