To develop sustainable strategies for mitigating ruminal methanogenesis and improving nitrogen efficiency in dairy systems, this study investigated how low-dose tannic acid (T), tea polyphenols (TP), and their combination (T+TP; 50:50) modulate rumen microbiota and function. A sample of Holstein cows were given four dietary treatments: (1) control (basal diet); (2) T (basal diet + 0.4% DM tannic acid); (3) TP (basal diet + 0.4% DM tea polyphenols); and (4) T+TP (basal diet + 0.2% DM tannic acid + 0.2% DM tea polyphenols). We comprehensively analyzed rumen fermentation, methane production, nutrient digestibility, milk parameters, and microbiota dynamics. Compared with the control group, all diets supplemented with additives significantly reduced enteric methane production (13.68% for T, 11.40% for TP, and 10.89% for T+TP) and significantly increased milk protein yield. The crude protein digestibility significantly increased in the T group versus control. The results did not impair rumen health or fiber digestion. Critically, microbiota analysis revealed treatment-specific modulation: the T group showed decreased Ruminococcus flavefaciens abundance, while all tannin treatments reduced abundances of Ruminococcus albus and total methanogens. These microbial shifts corresponded with functional outcomes—most notably, the T+TP synergy drove the largest reductions in rumen ammonia-N (34.5%) and milk urea nitrogen (21.1%). Supplementation at 0.4% DM, particularly the T+TP combination, effectively enhances nitrogen efficiency and milk protein synthesis while reducing methane emissions through targeted modulation of key rumen microbiota populations, suggesting potential sustainability benefits linked to altered rumen fermentation.

This study evaluated the effects of rumen-protected chromium-nicotinic acid (RPCNA) supplementation on lactation performance, nutrient digestion, ruminal fermentation, serum biochemical parameters and antioxidant in lactating water buffaloes under conditions of a critical thermal comfort index (TCI). Healthy lactating water buffaloes (milk yield = 5.96 ± 0.21 kg; parity = 2.96 ± 0.15, mean ± SD) were randomly assigned to four groups, which were the control (without RPCNA), and three treatment groups designated as RPCNA2, RPCNA4, and RPCNA6, with 0, 2, 4, and 6 mg/(d·head) of RPCNA, respectively. During a 56-day experimental period, the average temperature was 29 °C with an average TCI of 40.60, indicating a critical state of environmental stress. The results showed that the addition of RPCNA to the diet had no significant effect on the apparent digestibility of nutrients, lactation performance, and dry matter intake in lactating water buffaloes. However, RPCNA supplementation positively influenced rumen fermentation, reducing ammonia nitrogen (NH₃-N) concentrations and promoting microbial protein synthesis. Supplementation with RPCNA4 and RPCNA6 decreased rumen NH₃-N levels by 48.3% and 36.4%, respectively, while RPCNA4 increased isobutyrate concentrations. This demonstrates that the coating technology does not provide 100% rumen protection. Serum analysis revealed that RPCNA significantly increases total antioxidant capacity (T-AOC). Additionally, the supplementation of 4 mg/(d·head) of RPCNA, improved serum glutathione peroxidase (GSH-Px) activity and reduced malondialdehyde (MDA) levels. These findings suggest that moderate doses of RPCNA supplementation can improve antioxidant status and rumen nitrogen metabolism in lactating water buffaloes under critical state, without significantly altering milk production or composition.