This study aimed to evaluate the effects of Pleurotus ostreatus spent mushroom substrate (P.SMS), which is characterized by high production but low utilization, on feed utilization and liver serum indices from the perspective of duodenal microorganisms. Forty-five 3-month-old Hu sheep were randomly assigned to five groups and fed diets in which whole-plant corn silage (WPCS) was substituted with P.SMS at levels of 0% (Con), 5% (PSMS5), 10% (PSMS10), 15% (PSMS15), or 20% (PSMS20). The results indicated that the addition of P.SMS complexly influenced the apparent digestibility of dry matter, organic matter, and crude protein, with PSMS10 showing the highest digestibility of these nutrients. P.SMS inclusion significantly affected serum alanine aminotransferase levels, with PSMS5 showing higher levels than both the Con and PSMS20 groups (p < 0.05). Importantly, the inclusion of P.SMS did not affect the richness and diversity of duodenal microorganisms. Significant differences in the phyla Verrucomicrobiota and Spirochaetota were observed between the Con and PSMS20 groups. The observed trend towards an increase in the genus Trichoderma (p = 0.057) suggests that P.SMS is susceptible to contamination by this genus, which in turn affects the structure of the intestinal flora. Furthermore, functional gene predictions indicated differences in amino acid metabolism among the groups (p < 0.05). In conclusion, feeding with 10% P.SMS resulted in the highest digestibility without adversely affecting the structure of the duodenal community or liver function.

This study evaluates the effects of a compound microecological preparation named ATABG, which is composed of antimicrobial peptide ID13 and Saccharomyces boulardii, on Hu sheep’s growth performance, feed digestibility, and rumen parameters. A total of 40 three-month-old Hu sheep (21.65 ± 0.33 kg) were randomly assigned to two groups: the control group (Con), which received a basal diet, and the experimental group (ATABG), which received the same diet supplemented with 1 g/kg ATABG on a dry matter basis. After a 10-day pre-feeding period to adapt the animals to the experimental diet, dry matter intake and weight gain were recorded during the subsequent 63-day trial period. Body weight was measured on days 1, 21, 42, and 63 of the trial, and animals were slaughtered on day 63 to collect rumen fluid and tissue. Results indicated that ATABG supplementation significantly increased the apparent digestibility of crude protein, neutral detergent fiber, acid detergent fiber, and organic matter (p < 0.05). Rumen fluid analysis revealed increased microbial protein concentration and cellulase activity (p < 0.05) in the ATABG group. Microbiota analysis indicated that ATABG increased the relative abundance of Ruminococcus and Proteobacteria, elevated Firmicutes, and reduced Bacteroidota (p < 0.05). Correlation analysis showed Ruminococcus was positively associated with crude protein digestibility, while Quinella correlated with growth-related indices (r > 0.4, p < 0.05). In conclusion, ATABG supplementation improves protein digestibility and rumen microbial protein synthesis by enriching Ruminococcus and enhancing cellulase activity, potentially optimizing nitrogen utilization in Hu sheep.

Machine-harvested cotton residue is a major byproduct of cotton processing whose utilization is limited by a lignocellulosic matrix and antinutritional factors. This study combined an L49(76) orthogonal design with entropy-weighted TOPSIS for multi-indicator selection and then used Plackett–Burman screening and Box–Behnken response surface methodology (RSM) to optimize NDF degradability as a response-specific objective. Range analysis showed that fermentation time was the dominant factor for several fiber-related indices. TOPSIS selected A₄B₃C6D2E6F1 as the preferred multi-indicator treatment, corresponding to 15% glucose, 10% urea, 65% moisture, 14 days, a bacterial ratio of Enterococcus faecium: Bacillus licheniformis: Bacillus amyloliquefaciens = 15:10:5, and an enzyme ratio of cellulase: laccase: lignin peroxidase = 5:10:15. Relative to the raw residue, NDF, cellulose, hemicellulose, and ADL contents decreased by 25.83%, 43.54%, 74.40%, and 45.65%, respectively. In the subsequent NDF-oriented RSM stage, PBD identified fermentation time, urea, and glucose as the three quantitative factors taken forward to BBD. The model predicted an NDF degradability of 39.054% at 20.449 days, 9.865% urea, and 9.901% glucose; a rounded operating point of 20 days, 10% urea, and 10% glucose yielded a mean of 37.03% in three replicate runs. Thus, TOPSIS and RSM provided complementary results for different optimization objectives rather than a single global optimum.

Objective: This study systematically evaluated the nutritional compositions and bioactive compounds of six unconventional feed resources (Pepper residue (PR), Grape marc (MC), Pepper straw (PS), Lycium barbarum branches and leaves (LBBL), Licorice straw (LS), and Cyperus esculentus leaves (CES)). It also assessed the rumen degradability and rumen fermentation characteristics at different substitution levels through in vitro and in situ methods, to explore their potential application in sheep diets. Methods: Samples were analyzed considering nutrient composition, amino acids, polyunsaturated fatty acids (PUFAs), and bioactive compounds. In situ degradation was measured using rumen-fistulated sheep, and in vitro batch fermentation culture was conducted at varying substitution levels (0–100%) to measure gas production, pH, VFAs, NH₃-N, and microbial crude protein (MCP). Results: The six unconventional feed resources showed significant differences in nutrient composition, bioactive compounds, and fermentation performance. Crude protein (CP) ranged from 4.45% to 15.76%, with LS highest in total amino acids. LBBL contained 4.24 g/kg Lycium barbarum polysaccharides, LS had 9.24 g/kg liquiritin, GM was richest in proanthocyanidins, and PS had more capsaicin than PR. PR exhibited the highest DM degradation (74.77%, p < 0.001), followed by LS; CEL was lowest. PR and LS also had the highest CP degradation. In vitro fermentation revealed significant differences in fermentation characteristics among the six feeds. At 100% replacement, PR and LS exhibited high cumulative gas production, elevated MCP concentrations, and total VFAs of 54.41 and 64.02 mmol/L (p < 0.001), respectively. At 25% replacement, GM and CEL achieved high concentrations of VFAs and maintained MCP levels of 27.84 and 31.57 mg/dL (p < 0.001). PS reached its maximum total VFAs and MCP at 50% replacement, while LBBL reached 64.90 mmol/L total VFAs and 32.63 mg/dL MCP at 75% replacement. Conclusions: Nutrient composition and degradation kinetics varied significantly among substrates. PR had the highest DM degradability, while CEL had the lowest. PR and LS maintained stable fermentation at 100% substitution. GM and CEL were most effective at 25%; PS at 50%; and LBBL at 75% substitution levels.