Corn and soybean meal (SBM) are two of the most common feed ingredients used in pig feeds. However, a variety of antinutritional factors (ANFs) present in corn and SBM can interfere with the bioavailability of nutrients and have negative health effects on the pigs. In the present study, two-stage fermentation using Bacillus subtilis followed by Enterococcus faecium was carried out to degrade ANFs and improve the nutritional quality of corn and SBM mixed feed. Furthermore, the microbial composition and in vitro nutrient digestibility of inoculated mixed feed were determined and compared those of the uninoculated controls.

Our results suggest that two-stage fermentation using B. subtilis followed by E. faecium is an effective approach to improve the quality of corn-soybean meal mixed feed.

During the fermentation process, B. subtilis and lactic acid bacteria (LAB) were the main dominant bacteria in the solid-state fermented inoculated feed, and fermentation produced a large amount of lactic acid (170 mmoL/kg), which resulted in a lower pH (5.0 vs. 6.4) than the fermented uninoculated feed. The amounts of soybean antigenic proteins (β-conglycinin and glycinin) in mixed feed were significantly decreased after first-stage fermentation with B. subtilis. Inoculated mixed feed following two-stage fermentation contained greater concentratioin of crude protein (CP), ash and total phosphorus (P) compared to uninoculated feed, whereas the concentrations of neutral detergent fiber (NDF), hemicellulose and phytate P in fermendted inoculated feed declined (P < 0.05) by 38%, 53%, and 46%, respectively. Notably, the content of trichloroacetic acid soluble protein (TCA-SP), particularly that of small peptides and free amino acids (AA), increased 6.5 fold following two-stage fermentation. There was no difference in the total AA content between fermented inoculated and uninoculated feed. However, aromatic AAs (Phe and Tyr) and Lys in inoculated feed increased, and some polar AAs, including Arg, Asp, and Glu, decreased compared with the uninoculated feed. In vitro dry matter and CP digestibility of inoculated feed improved (P < 0.05) compared with the uninoculated feed. Conclusions: Our results suggest that two-stage fermentation using B. subtilis followed by E. faecium is an effective approach to improve the quality of corn-soybean meal mixed feed.