Histone deacetylase (HDAC) inhibition attenuates inflammation in rodents and short-chain fatty acids (SCFAs) are effective HDAC inhibitors. Therefore, the objective of this study was to evaluate the role of the SCFAs sodium propionate (SP) and sodium butyrate (SB) as HDAC-dependent regulators of inflammatory gene expression in bovine mammary epithelial cells (MAC-Ts). We postulated that SP and SB would decrease inflammation in MAC-Ts by inhibiting HDAC activity and increasing histone H3 acetylation and consequently decreasing inflammatory gene expression. For this study, MAC-Ts stimulated with lipopolysaccharide (LPS) were used as a model for bovine mammary epithelial cell inflammation. MAC-Ts were cultured in a basal medium. Cell lysates were incubated with SP or SB (0 to 5 mM) for 2 h prior to HDAC substrates incubation for an additional 2 h and HDACs activity was determined. Next, cells were pretreated with SP or SB (0 to 3.0 mM) for 2 h prior to LPS (1 µg/mL) stimulation for an additional 2 h and assessed for histone H3 acetylation. Then, cells were pretreated with SP or SB (1 mM) for 24 h prior to LPS (1 µg/mL) stimulation for an additional 2 h and RNA was isolated for inflammatory gene expression evaluation by PCR array and gene validation was performed using quantitative real-time PCR. One-way ANOVA followed by Tukey post hoc analysis was conducted and statistical significance set at P < 0.05. SP and SB concentration-dependently and selectively inhibited class I HDAC activity, which differed between SCFAs, where SB inhibited (P < 0.05) HDACs 2, 3, and 8, while SP inhibited (P < 0.05) HDACs 2 and 8. Histone H3 acetylation was concentration-dependently increased by SCFAs and likewise the differential regulation of HDAC activity, SCFAs effected differently histone H3 acetylation, where SB increased (P < 0.05) H3K9/14, H3K18 and H3K27 acetylation, while SP increased (P < 0.05) H3K9/14 and H3K18 acetylation. However, SCFAs did not decrease (P > 0.05) overall inflammatory gene expression. Under our experimental conditions, findings suggest that in MAC-Ts, SCFAs regulate epigenetic markers on nucleosomal DNA in addition to regulation of inflammatory gene events independent of HDAC activity. Nevertheless, examination of SCFAs and/ or HDACs inhibitors in bovine mammary gland is worth being further investigated to delineate the potential impact of HDAC inhibition and histones hyperacetylation on mammary gland tissue inflammation.

The objectives of this study were: 1) to compare the effects of live yeast (LY), yeast fermentation product (YFP), a mix of Lactobacillus acidophilus and Propionibacterium freudenreichii (MLP), and Lactobacillus plantarum included as additives in dairy cows’ diets on in vitro ruminal fermentation and gas production (GP); and 2) to evaluate the effects of L. plantarum as direct-fed microbials (DFM) in dairy cows’ diets on in vitro ruminal fermentation, GP, nutrient digestibility, and N metabolism. Three experiments were carried out: Exp. 1 had the objective to compare all additives regarding ruminal fermentation parameters: an Ankom GP system was used in a completely randomized design, consisting of four 48 h incubations, and eight replications per treatment. There were eight treatments: a basal diet without additive (CTRL) or with one of the following additives: LY, YFP, MLP, or L. plantarum at four levels (% of diet Dry Matter (DM)): 0.05% (L1), 0.10% (L2), 0.15% (L3), and 0.20% (L4). In Exp. 2, a batch culture was used to evaluate ruminal fermentation, and CO2 and CH4 production using the same treatments and a similar experimental design, except for having 16 replications per treatment. Based on Exp. 1 and 2 results, Exp. 3 aimed at evaluating the effects of the L. plantarum on ruminal true nutrient digestibility and N utilization in order to evaluate the use of L. plantarum as DFM. The treatments CTRL, MLP, L1, and L2 were used in a replicated 4 × 4 Latin square design using a dual-flow continuous culture system. Data were analyzed using linear and nonlinear regression; treatment means were compared through contrasts, and L treatments in Exp. 1 and 2 were tested for linear, quadratic, and cubic effects. In Exp. 1, all treatments containing additives tended to reduce OM digestibility as well as reduced total volatile fatty acids (VFA) concentration and total GP. The YFP had greater OM digestibility than LY, and MLP treatment had greater total VFA concentration compared to L. plantarum treatments. In Exp. 2, additives reduced CO2 production, and there were no major differences in CH4. In Exp. 3, all additives reduced NH3-N concentration. In conclusion, pH and lactate concentration were not affected in all three experiments regardless of additive tested, suggesting that these additives may not improve ruminal fermentation by pH modulation; and L. plantarum may improve ruminal N metabolism when used as DFM in high-producing dairy cows’ diets, mainly by reducing NH3-N concentration.

-The objective of this study was to determine the optimal level of dried brewers' grains (DBG) to replace soybean meal in diets for lactating Holstein cows. Five cows, around 88±28 days in milk, were distributed in a 5 × 5 Latin square design and fed diets containing different levels of DBG (0, 25, 50, 75, and 100%). The forage:concentrate ratio of the diet was 50:50. Feed intake, dry matter, nutrient digestibility, microbial synthesis, milk production and composition, and the economic viability of the diets were evaluated. There was reduction in dry matter intake and, consequently, in crude protein and non-fiber carbohydrates with increased levels of DBG. This occurred due to physical limitation of rumen caused by increased neutral detergent fiber intake. Ether extract intake also increased with levels of DBG due to higher concentrations of this nutrient in the diet. Apparent dry matter, organic matter, ether extract, and neutral detergent fiber digestibility increased with replacement of soybean meal by DBG. Milk production showed a quadratic effect and the levels of fat, protein, and total solids reduced linearly. Each 1% of soybean meal replaced by DBG in concentrate led to a reduction of 0.04, 0.02, and 0.06 g kg -1 of milk fat, protein, and total solids, respectively. The milk production efficiency increased linearly and the microbial synthesis efficiency was not affected. The economic return increased along with the DBG levels. Thus, DBG levels replacing up to 75% of soybean meal can be used to feed lactating cows, since it provides improvements in digestibility, milk production efficiency, and economic return without affecting microbial efficiency.Key Words: by-product, digestibility, intake, milk composition, protein, purine Revista Brasileira de Zootecnia

The objective of this study was to evaluate the effect of inclusion of condensed tannins (CT) from black wattle (Acacia mearnsii) on feed intake, ruminal protozoa population, ruminal fermentation, and nutrient digestibility in Jersey steers. Five ruminally-cannulated steers were used in a 5 × 5 Latin square design, with five periods of 20 days each (14 days for diet adaptation and six days for sample collection per period). Treatments were composed of dietary inclusion levels of condensed tannins at 0, 5, 10, 15, and 20 g/kg of diet dry matter. Intakes of dry matter, organic matter, ether extract, crude protein, neutral detergent fiber, and total digestible nutrients were not affected by condensed tannins. The ruminal pH was reduced linearly with tannin levels. Ruminal ammonia nitrogen concentration was not affected by tannins. Tannins reduced the molar proportion of acetate and did not affect the ruminal protozoal population, which might be related to the low doses used. Digestibilities of dry matter, organic matter, and neutral detergent fiber were not altered; however, there was a linear reduction in crude protein digestibility. Based on these results, CT extracts from black wattle are not recommended for improving nutrient utilization in steers at the tested levels.