Qiufeng Li

Ensiling high-moisture alfalfa with peanut vine not only avoids alfalfa nutrient loss during the wilting stage but also maximizes the use of agricultural waste peanut vine. The appropriate mixed ratio of high-moisture alfalfa and peanut vine has been studied in our previous study. However, the effect of additives on improving the nutrition and fermentation quality of the mixed silage of alfalfa and peanut vine has not been investigated. This study aimed to assess the adaptation and association of Lactiplantibacillus plantarum, cellulase and tannin in the mixed silage of alfalfa and peanut vine alone or in combination on fermentation quality, chemical composition, and microbial communities. The harvested fresh alfalfa and dry peanut vine were cut into 2 cm lengths by a crop chopper and they were thoroughly mixed at a ratio of 7:3. The mixtures were treated with no addition (CK), L. plantarum (Lp, 1 × 10⁶ CFU/g fresh weight), cellulase (Ce, 5 g/kg fresh weight), tannin (Ta, 40 g/kg dry matter), and their combinations (LpCe, LpTa, CeTa, LpCeTa). After 45 days of fermentation, silage treated with Lp, Ce, and Ta had lower pH and ammonia-N (NH₃-N) content and higher concentrations of lactic acid compared with the CK group. LpCeTa-treated silage inhibited protein degradation by reducing pH value and ammonia-N concentrations during ensiling processes. The LpCeTa group increased (p < 0.05) water-soluble carbohydrate (WSC) content and reduced (p < 0.05) acid detergent fiber and neutral detergent fiber contents in mixed silage. Furthermore, the LpCeTa group increased the relative abundance of Lactobacillus and decreased the relative abundance of Enterococcus and Weissella as compared with the CK group. Results of the current study indicated that the combined use of L. plantarum, cellulase, and tannin could serve as a promising strategy for the preservation of ensiling fresh alfalfa mixed with peanut vine and provide a reference for the re-utilization of by-product.

Fresh alfalfa is difficult to ensile successfully because of its high moisture content and greater susceptibility to spoilage by Clostridia, Bacilli or Enterobacter. In this study, we evaluated the effects of mixing high-moisture alfalfa with peanut vine in different proportions on the bacterial communities and fermentation characteristics of silage. The high-moisture alfalfa and peanut vine were mixed at ratios of 10:0 (CK), 8:2 (TI), 7:3 (T2), 6:4 (T3) and 5:5 (T4), respectively. For each treatment, silos (25 × 35 cm) were anaerobically fermented in darkness at room temperature and analyzed after 45 days. The results showed that the CK silage was weakly fermented, as indicated by a low lactic acid concentration, a high pH value, and high levels of propionic acid (PA), butyric acid (BA) and ammonia nitrogen (NH₃-N). As the proportion of peanut vine in the mixture increased, the pH level decreased, and levels of BA, propionic acid, NH₃-N, crude protein(CP), nonprotein nitrogen and soluble protein also declined (p < 0.05), while the lactic acid concentration increased and levels of neutral detergent fiber (NDF) and water-soluble carbohydrates (WSC) also rose (p < 0.05). A protein component analysis of silage mixtures using the Cornell Net Carbohydrate and Protein System (CNCPS) showed that the content of the nonprotein nitrogen component (PA) decreased when the proportion of peanut vine increased, whereas the content of rapidly degraded protein (PB1) increased. Mixing with peanut vine also influenced the distribution of the bacterial community. Compared with the CK silage, the relative abundances of Enterococcus, Garciella and Anaerosporobacter in T2, T3 and T4 were significantly lower, while the relative abundance of Lactobacillus was significantly higher. In the T2, T3 and T4 groups, Garciella and Anaerosporobacter were not detected. In summary, in this study, we ensiled high-moisture alfalfa, which was weakly fermented. We found that mixing with peanut vine improved fermentation quality and optimized the structure of the bacterial community. Therefore, to improve the fermentation quality and nutritional value of silage, high-moisture alfalfa should be ensiled with at least 30% peanut vine.

The aim of this study was to investigate the effect of low-protein diets supplemented with rumen-protected lysine (RPLys) and methionine (RPMet) on growth performance, rumen fermentation, blood biochemical parameters, nitrogen metabolism, and gene expression related to N metabolism in the liver of Holstein bulls. Thirty-six healthy and disease-free Holstein bulls with a similar body weight (BW) (424 ± 15 kg, 13 months old) were selected. According to their BW, they were randomly divided into three groups with 12 bulls in each group in a completely randomized design. The control group (D1) was fed with a high-protein basal diet (CP13%), while bulls in two low-protein groups were supplied a diet with 11% crude protein and RPLys 34 g/d·head + RPMet 2 g/d·head (low protein with low RPAA, T2) or RPLys 55 g/d·head + RPMet 9 g/d·head (low protein with high RPAA, T3). At the end of the experiment, the feces and urine of dairy bulls were collected for three consecutive days. Blood and rumen fluid were collected before morning feeding, and liver samples were collected after slaughtering. The results showed that the average daily gain (ADG) of bulls in the T3 group was higher than those in D1 (p < 0.05). Compared with D1, a significantly higher nitrogen utilization rate (p < 0.05) and serum IGF-1 content (p < 0.05) were observed in both T2 and T3 groups; however, blood urea nitrogen (BUN) content was significantly lower in the T2 and T3 groups (p < 0.05). The content of acetic acid in the rumen of the T3 group was significantly higher than that of the D1 group. No significant differences were observed among the different groups (p > 0.05) in relation to the alpha diversity. Compared with D1, the relative abundance of Christensenellaceae_R-7_group in T3 was higher (p < 0.05), while that of Prevotellaceae _YAB2003_group and Succinivibrio were lower (p < 0.05). Compared with D1 and T2 group, the T3 group showed an expression of messenger ribonucleic acid (mRNA) that is associated with (CPS-1, ASS1, OTC, ARG) and (N-AGS, S6K1, eIF4B, mTORC1) in liver; moreover, the T3 group was significantly enhanced (p < 0.05). Overall, our results indicated that low dietary protein (11%) levels added with RPAA (RPLys 55 g/d +RPMet 9 g/d) can benefit the growth performance of Holstein bulls by reducing nitrogen excretion and enhancing nitrogen efficiency in the liver.

The objective of this study was to investigate the effects of dietary rumen-degradable starch (RDS, g/kg of DM) to rumen-degradable protein (RDP, g/kg of DM) ratios (SPR) on in vitro rumen fermentation characteristics and microbial protein synthesis (MCPS). Treatments were eight diets with SPR of 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5 and 2.6 and were formulated to be isoenergetic, isonitrogenous, and isostarch. Substrates were anaerobically incubated in sealed culture vials (100 mL) for 6, 24 or 48 h. Three incubation runs were conducted within two consecutive weeks. With the increase of the dietary SPR, the gas production (GP), in vitro dry matter disappearance (IVDMD) and concentration of MCPS and total volatile fatty acids (TVFA) linearly increased after 6 h of incubation (p ≤ 0.01), whereas they quadratically increased and peaked at the SPR of 2.3 after 24 and 48 h of incubation (p < 0.05). In response to dietary SPR increasing, the in vitro neutral detergent fiber disappearance (IVNDFD) quadratically increased (p < 0.01), and the ammonia nitrogen (NH₃-N) concentration linearly decreased (p < 0.01) after 6, 24 and 48 h of incubation. Based on the presented results, an SPR of 2.3 is recommended for formulating a diet due to its greatest IVDMD, IVNDFD, GP, TVFA and MCPS. However, as the results obtained are strictly dependent on the in vitro conditions, further in vivo studies are needed to verify our findings.