β-sitosterol (BSS) is a plant-derived natural bioactive compound, its cellular mechanism of anti-inflammatory activity has been proven recently. Little information is available regarding the application of BSS on ruminants under high grain diet. The objective of this study was to evaluate the effects of dietary BSS supplementation on inflammatory response, ruminal fermentation characteristics and the composition of the ruminal bacterial community under high grain diet. Eight rumen-cannulated Hu sheep (59.7 ± 4.8 kg of initial body weight) were randomly assigned into a replicated 4 × 4 Latin square design trial. Sheep were fed a high grain diet (non-fiber carbohydrate: neutral detergent fiber = 2.03) supplemented either with 0.25 (LBS), 0.5 (MBS), 1.0 (HBS) or without (CON) g BSS /kg dry matter diet. On day 21 of each period, rumen content samples were obtained at 6 h postfeeding, and blood samples were obtained before morning feeding. The data showed that compared with control group, Dietary BSS supplementation decreased serum concentrations of tumor necrosis factor, interleukin (IL)-6, and IL-1β. The ruminal pH and acetate concentration for BSS treatment were improved, while concentration of propionate, butyrate and lactate was decreased. The result of Illumina MiSeq sequencing of 16S rRNA gene revealed that BSS addition can increase the proportion of Prevotella_1, Rikenellaceae_RC9_gut_group, Prevotella_7, and Selenomonas_1, and decrease the proportion of Lachnospiraceae_NK3A20_group. These results indicated that BSS attenuates high grain diet-induced inflammatory response and modifies ruminal fermentation. In addition, the BSS dietary supplementation at the level of 0.5 g/kg is recommended in sheep.

Aims:To explore the impact of ambient pH on lactate catabolism by Megasphaera elsdenii BE2-2083 and Selenomonas ruminantium HD4 in both pure culture and in binary mixed culture. Methods and results:The growth rate, substrate consumption, product formation, enzymatic activity and gene expression of M. elsdenii and S. ruminantium at various pHs were examined. Furthermore, the metabolism of lactate catabolism pathways for M. elsdenii and S. ruminantium in the co-culture system was investigated by chasing the conversion of sodium L-[3-13 C]-lactate in nuclear magnetic resonance. In the pure culture systems, ambient pH had significant effects on the growth of M. elsdenii, whereas S. ruminantium was less sensitive to pH changes. In addition, lactate metabolic genes and activities of key enzymes were affected by ambient pH in M. elsdenii and S. ruminantium. In the co-culture system, low ambient pH reduced the contribution lactate catabolism by M. elsdenii. Conclusion: M. elsdenii BE2-2083 and S. ruminantium HD4 lactate degradation affected by ambient pH. Significance and impact of the study: This study demonstrates the regulatory mechanisms of lactate decomposing bacteria in lactate catabolism under the condition of subacute ruminal acidosis.

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