| Figure 3. Effects of Aspergillus oryzae and Aspergillus niger co-cultivation fermentation extract on
starch disappearance in total mixed ration and corn silage at 7 h in vitro incubation. AOAN indi-
cates fermentation with Aspergillus oryzae and Aspergillus niger co-cultivation fermentation extract.
*, p < 0.05; **, 0.05 < p < 0.1; TMR, total mixed ration.
3.3. Fermentation Parameters
Figure 4 shows the effects of AOAN supplementation on fermentation parameters.
The pH values were decreased (Figure 4A; p < 0.05), and NH
3
-N concentration in TMR
was decreased with AOAN (Figure 4B; p < 0.05). The total VFAs were increased or tented
to be increased by AOAN (Figure 4C). There were no differences in the molar proportions
of propionate and butyrate (Figure 4E,F; p > 0.05). AOAN increased the molar proportion
of acetate, and the acetate to propionate ratio of oat hay and alfalfa hay (Figure 4D,H; p <
0.05). The molar proportion of total branched fatty acid including isobutyric and isovaleric
was not affected by treatment (Figure 4G; p > 0.05).
3.4. Bacterial Community
3.4.1. Sequence Depth, Diversity, and Composition
After data filtering, a total of 2,263,946 reads from 32 samples were obtained with a
mean of 70,748 reads for individual samples. The Good’s coverage for each sample was
deemed sufficient, with values > 99.00% for all bacterial communities, implying that the
current sequencing depth was sufficient to be representative of the microbiota (Supple-
mental Figure S2. The sample-based rarefaction curves in Supplemental Figure S3 indicate
that our sequencing depth was sufficient to accurately describe the bacterial composition
of all groups.
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