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Simple Summary  The rumen is a large fermentation chamber that enables dairy cows to utilize otherwise indigestible plant polymers and compounds for their nutrition, highlighting a crucial difference between ruminants and monogastrics. The key point in this process is the pool of enzymes secreted by microorganisms. Hence, exogenous enzymes from additives are important for high-production dairy cows to improve the utilization of feeds. In this study, we used the  Aspergillus oryzae  and  Aspergillus niger  co-cultivation extract (AOAN) to improve the digestibility of roughage and total mixed ration in vitro. Our results indicated that the digestibility of nutrients in feeds was significantly increased by AOAN supplementation, except for the digestibility of crude protein in the total mixed ratio (TMR). Furthermore, the diversity of the bacteria in TMR and oat hay was increased with AOAN supplementation. Although broad effectiveness of AOAN was established, regardless of roughage types, the mechanism may be different according to roughage types.    Abstract  AOAN may provide enzymes to improve the digestibility of feeds and enhance rumen fermentation. This study determined the effects of AOAN on digestibility, fermentation characteristics, and bacterial composition using in vitro gas recording fermentation system. A total of 30 mg of AOAN was supplemented into 500 mg of TMR, corn silage, oat hay, and alfalfa hay. Fermentation parameters and bacterial communities were determined after 48 h fermentation, and digestibility was determined after 7, 24, 30, and 48 h fermentation. Gas production and dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), and acid detergent fiber (ADF) digestibility were significantly increased by AOAN supplementation at 48 h ( p  < 0.05), except for digestibility of CP of the TMR ( p  > 0.05). AOAN increased starch digestibility in corn silage ( p  < 0.05) and tended to increase that in TMR (0.05 <  p  < 0.10). AOAN supplementation increased total volatile fatty acid production ( p  < 0.05). The molar proportions of acetate and acetate to propionate ratio of oat hay and alfalfa hay were increased ( p  < 0.05). The 16S rRNA analysis revealed that the microbial richness of TMR and oat hay, and microbial evenness of TMR were increased ( p  < 0.05). AOAN did not affect the α diversity, β diversity, and bacterial composition of the corn silage. The relative abundance of  Prevotella  was increased and  Ruminococcus  was decreased in TMR, oat hay, and alfalfa hay. In conclusion, results suggest that AOAN has the potential to improve the utilization of diets differently, including providing enzymes with changing microbiota (TMR, oat hay, and alfalfa hay) or providing enzymes alone (corn silage).

animals

Aspergillus oryzae and Aspergillus niger Co-Cultivation Extract Affects In Vitro Degradation, Fermentation Characteristics, and Bacterial Composition in a Diet-Specific Manner