The Effects of Dietary Resveratrol and β-Hydroxy-β-Methylbutyric Acid Supplementation at Two Protein Levels on the Ruminal Microbiome and Metabolome of Tibetan Sheep

Resveratrol (RES) and β-hydroxy-β-methylbutyric acid (HMB) have antioxidant, anti-inflammatory, and other beneficial properties. Here, we hypothesize that supplementation with RES and HMB could affect the rumen function in Tibetan sheep. This study aims to explore the effects of RES and HMB supplementation at different protein levels on the rumen microbial and metabolite compositions of Tibetan sheep. Four treatments (n = 30) were prepared according to a 2 × 2 factorial arrangement, with two dietary protein levels (12% and 14%) and two feed additives (RES 1.50 g/day and HMB 1.25 g/day). The experimental treatments were fed diets with 12% CP level non-supplemented (L), 12% protein with RES and HMB (L-RES-HMB), 14% CP level non-supplemented (H), and 14% protein with RES and HMB (H-RES-HMB), respectively. Our results indicated that the trypsin, chymotrypsin, and lipase were significantly increased in the H-RES-HMB group ( p < 0.05), while the lipopolysaccharide (LPS) concentration was significantly reduced ( p < 0.05). The alpha diversity analysis found that the ACE indices of the L-RES-HMB, and H-RES-HMB groups was significantly higher than that of the L group ( p < 0.05). Additionally, compared with the L, L-RES-HMB, and H groups, it was found that the abundance of Euryarchaeota , Spirochaeota , and Metanobrevibacter was significantly increased in the H-RES-HMB group, while the abundance of Proteobacteria was significantly decreased ( p < 0.05). A total of 745 significantly different metabolites were identified, of which 14 metabolites were common among the three comparative groups. Differential metabolites were mainly enriched in pathways including the pyrimidine metabolism, the glycine, serine, and threonine metabolisms, and ABC transporters. Overall, CP level and RES/HMB exhibited positively interaction effect on digestive enzyme activity and antioxidant capacity. Dietary RES and HMB supplementation on 14% CP level improved the ruminal digestive enzyme activity and antioxidant capacity through modulating the microbial community and regulating the metabolism.