M. Oba

BOARD-INVITED REVIEW: The hepatic oxidation theory of the control of feed intake and its application to ruminants

Feed and energy intake of ruminant animals can change dramatically in response to changes in diet composition or metabolic state, and such changes are poorly predicted by traditional models of feed intake regulation. Recent work suggests that temporal patterns of fuel absorption, mobilization, and metabolism affect feed intake in ruminants by altering meal size and frequency. Research with nonruminants suggests that meals can be terminated by signals carried from the liver to the brain via afferents in the vagus nerve and that these signals are affected by hepatic oxidation of fuels and generation of ATP. We find these results consistent with the effects of diet on feed intake of ruminants. Of fuels metabolized by the ruminant liver, propionate is likely a primary satiety signal because its flux to the liver increases greatly during meals. Propionate is utilized for gluconeogenesis or oxidized in the liver and stimulates oxidation of acetyl CoA. Although propionate is extensively metabolized by the ruminant liver, there is little net metabolism of acetate or glucose, which may explain why these fuels do not consistently induce hypophagia in ruminants. Lactate is metabolized in the liver but has less effect on satiety, probably because of greater latency for reaching the liver within meals and because of less hepatic extraction compared with propionate. Hypophagic effects of fatty acid oxidation in the liver are likely from delaying hunger rather than promoting satiety because beta-oxidation is inhibited during meals by propionate. A shortage of glucose precursors and increased fatty acid oxidation in the liver for early lactation cows lead to a lack of tricarboxylic acid (TCA) cycle intermediates, resulting in a buildup of the intracellular acetyl-CoA pool and export of ketone bodies. In this situation, hypophagic effects of propionate are likely enhanced because propionate entry into the liver provides TCA cycle intermediates that allow oxidation of acetyl-CoA. Oxidizing the pool of acetyl-CoA rather than exporting it increases ATP production and likely causes satiety despite the use of propionate for glucose synthesis. A better understanding of metabolic regulation of feed intake will allow diets to be formulated to increase the health and productivity of ruminants.

Epithelial Capacity for Apical Uptake of Short Chain Fatty Acids Is a Key Determinant for Intraruminal pH and the Susceptibility to Subacute Ruminal Acidosis in Sheep

Subacute ruminal acidosis (SARA) is a common digestive disorder occurring in ruminants, with considerable variation in the severity of SARA observed among animals fed the same diet. Our aim in this study was to determine whether differences in the capacity of the ruminal epithelium for the apical uptake of acetate and butyrate (determined in Ussing chambers after slaughter) explains differences observed for the severity of a preceding episode of SARA in vivo. Adult sheep with an indwelling small ruminant ruminal pH measurement system (SRS) were randomly assigned to either a SARA induction treatment (oral drench containing 5 g glucose/kg body weight; n = 17) or a sham treatment (SHAM; n = 7; 12 mL water/kg body weight). Sheep receiving the glucose drench were further classified as nonresponders (NR; n = 7) or responders (RES; n = 7) according to their ruminal pH profile for the 3 h following the oral drench. Mean ruminal pH for the 3 h following the drench differed among groups (P < 0.001), with it being highest for SHAM (6.67 +/- 0.08), intermediate for NR (5.97 +/- 0.05), and lowest for RES (5.57 +/- 0.08) sheep. The apical uptake of acetate and butyrate did not differ between SHAM and RES sheep. However, NR sheep had greater in vitro apical uptake of acetate and butyrate and a higher plasma beta-hydroxybutyrate concentration than RES sheep, suggesting greater absorptive capacity for NR. Differences between NR and RES were attributed to greater bicarbonate-independent, nitrate-sensitive uptake of acetate (P = 0.007), a tendency for greater bicarbonate-dependent uptake of acetate (P = 0.071), and greater bicarbonate-independent uptake of butyrate (P = 0.022). These data indicate that differences in the rates and pathways for the uptake of acetate and butyrate explain a large proportion of the individual variation observed for the severity of SARA.

Effects of sampling location and time, and host animal on assessment of bacterial diversity and fermentation parameters in the bovine rumen

Aims:  To investigate, using culture‐independent methods, whether the ruminal bacterial structure, population and fermentation parameters differed between sampling locations and time.

Effect of dietary forage to concentrate ratio on volatile fatty acid absorption and the expression of genes related to volatile fatty acid absorption and metabolism in ruminal tissue.

The objective of the study was to investigate the fractional rate of volatile fatty acid (VFA) absorption and the expression of genes encoding for transporters and enzymes involved in the absorption and metabolism of VFA in ruminal tissue when cattle were fed high or low concentrate diets. Twelve ruminally cannulated Holstein cows were used in a randomized complete block design. The low concentrate (LC) and high concentrate (HC) diets contained 8 and 64% dietary concentrate (dry matter basis), respectively. Cows were fed their respective diet for at least 28 d, following which data and samples were collected over 6 d. Ruminal pH was measured continuously for 72 h, and the in vivo VFA absorption and passage rates were measured using Co-EDTA and n-valeric acid as markers. Ruminal tissue was collected postslaughter from the ventral sac of the rumen, and gene expression was evaluated using quantitative real-time PCR. Dry matter intake was not affected by treatment, averaging 14.9 kg/d, but cows fed HC had lower mean ruminal pH (6.03 vs. 6.48), and a greater duration (376 vs. 10 min/d) that ruminal pH was <5.8. Ruminal VFA concentration was 24 mM higher for cows fed HC compared with LC; however, the fractional rate of VFA absorption and passage from the rumen was not affected by dietary treatment, averaging 23.4 and 9.6%/h, respectively. The expression of genes encoding for enzymes involved in VFA activation and ketogenesis were not affected by treatment. Cows fed HC tended to have a relative abundance of pyruvate dehydrogenase lipoamide alpha 1 mRNA transcripts that was 1.4 times lower than that of cows fed LC, but other enzymes involved in pyruvate metabolism or regulation of the citric acid cycle were not affected. Collectively, these results suggest that the dietary forage to concentrate ratio does not affect the fractional rate of VFA absorption in vivo, but potentially alters energy metabolism in ruminal tissue.

Increasing dietary sugar concentration may improve dry matter intake, ruminal fermentation, and productivity of dairy cows in the postpartum phase of the transition period

The current study was undertaken to investigate the effect of feeding diets varying in sugar concentration to postpartum transition cows on productivity, ruminal fermentation, and nutrient digestibility. We hypothesized that the high-sugar diet would increase dry matter intake and lactation performance. The secondary objective was to characterize changes in ruminal fermentation and nutrient digestibility over the first 4 wk of lactation. Fifty-two Holstein cows, including 28 primiparous and 24 multiparous cows, 10 of which were previously fitted with a ruminal cannula, were assigned to the experimental diets containing either high sugar (HS = 8.4%) or low sugar (LS = 4.7%) immediately after calving, based on their expected calving date. Data and samples were collected on d 5.2 +/- 0.3, 12.2 +/- 0.3, 19.2 +/- 0.3, and 26.1 +/- 0.3 relative to parturition for wk 1, 2, 3, and 4 respectively. Cows fed HS had increased dry matter intake compared with those fed LS (18.3. vs. 17.2 kg/d). Further, cows fed HS sorted for particles retained on the pan of the Penn State Particle Size Separator to a greater extent than cows fed LS. Feeding HS tended to increase nadir (5.62 vs. 5.42), mean (6.21 vs. 6.06), and maximum pH (6.83 vs. 6.65). The duration (h/d) and area (pH x min/d) that ruminal pH was below pH 5.8 were not affected by treatment. Ruminal volatile fatty acid concentration and molar proportions of individual volatile fatty acids were not affected by treatment. The digestibility of dry matter, organic matter, neutral detergent fiber, and starch were not affected by treatment, averaging 63.3, 65.2, 43.2, and 93.5%, respectively. Feeding HS decreased plasma glucose concentration compared with feeding LS (51.3 vs. 54.0 mg/dL), but concentration of plasma insulin was not affected by treatment, averaging 4.17 microIU/mL. Cows fed HS had higher concentrations of plasma beta-hydroxybutrate (17.5 vs. 10.5 mg/dL) and nonesterified fatty acids (344 vs. 280 microEq/L). Milk yield and milk composition were not affected by treatment, but a tendency for increased milk fat yield was observed for cows fed HS compared with LS (1.44 vs. 1.35 kg/d). The results of the current study imply that replacing cracked corn grain with sucrose may improve dry matter intake, ruminal pH status, and lactation performance.

Variation of bacterial communities and expression of Toll-like receptor genes in the rumen of steers differing in susceptibility to subacute ruminal acidosis

In order to determine differences in the ruminal bacterial community and host Toll-like receptor (TLR) gene expression of beef cattle with different susceptibility to acidosis, rumen papillae and content were collected from acidosis-susceptible (AS, n=3) and acidosis-resistant (AR, n=3) steers. The ruminal bacterial community was characterized using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) and quantitative real time PCR (qRT-PCR) analysis. Global R analysis of bacterial profile similarity revealed that bacterial diversity was significantly different between AR and AS groups for both rumen content (P=0.001) and epithelial (P=0.002) communities. The copy number of total bacterial 16S rRNA genes in content of AS steers was 10-fold higher than that of AR steers, and the copy number of total 16S rRNA genes of epimural bacteria in AR steers was positively correlated with ruminal pH (r=0.59, P=0.04), and negatively correlated with total VFA concentration (r=-0.59, P=0.05). The expressions of host TLR2 and 4 genes were significantly higher in AR steers compared to those in AS steers. These findings enhance our understanding about the ruminal microbial ecology and host gene expression changes that may be useful in the prevention of ruminal acidosis.

Relationship between rumen methanogens and methane production in dairy cows fed diets supplemented with a feed enzyme additive

Aims:  To investigate the relationship between ruminal methanogen community and host enteric methane (CH4) production in lactating dairy cows fed diets supplemented with an exogenous fibrolytic enzyme additive.

A single mild episode of subacute ruminal acidosis does not affect ruminal barrier function in the short term.

Twenty-four German Merino sheep (72.3±10.1 kg of body weight) were fed an all-hay diet and assigned to either the subacute ruminal acidosis (SARA) treatment (n=17) or sham treatment (n=7). The SARA sheep were orally dosed with a 2.2 M glucose solution to supply 5 g of glucose/kg of body weight, whereas sham sheep received an equal volume of water. Ruminal pH was measured for 48 h before and 3 h after the oral dose. Sheep were then killed and ruminal epithelia from the ventral sac were mounted in Ussing chambers. The serosal-to-mucosal flux rate of partially (3)H-labeled mannitol (J(mannitol-SM)), an indicator of barrier function, was measured while epithelia were exposed to 3 sequential in vitro measurement periods lasting 1 h each. The measurement periods consisted of baseline, challenge, and recovery periods and were interspersed by 30-min periods for treatment equilibration. Baseline conditions were pH 6.1 (mucosal solution) and pH 7.4 (serosal solution) with a bilateral osmolarity of 293 mOsm/L. During the challenge period, the mucosal side of the epithelia was exposed to either an acidotic challenge (pH 5.2, osmolarity 293 mOsm/L) or an osmotic challenge (pH 6.1, osmolarity 450 mOsm/L); a third group served as control (pH 6.1, osmolarity 293 mOsm/L). The mucosal buffer solution was replaced for the recovery period. In vivo, sheep on the SARA treatment had lower mean (5.77 vs. 6.67) and nadir (5.48 vs. 6.47) ruminal pH for the 3h following the oral drench compared with sham sheep, indicating the successful induction of SARA with the oral glucose dose. Despite the marked reduction in pH in vivo, induction of SARA had no detectable effects on the baseline measurements of J(mannitol-SM), tissue conductance (G(t)), and short-circuit current (I(sc)) in vitro. However, reducing mucosal pH to 5.2 in vitro had negative effects on epithelial barrier function in the recovery period, including increased J(mannitol-SM), increased G(t), and decreased I(sc). The osmotic challenge increased J(mannitol-SM) and G(t) and decreased I(sc) during the challenge period, which was reversible in the recovery period except for slight reduction in I(sc). Interactions between the in vitro treatment and measurement period were detected for J(mannitol-SM), G(t), and I(sc). These data indicate that a mild episode of SARA (nadir pH, 5.48; duration ruminal pH <5.8, 111 min relative to the 180-min measurement period) does not affect ruminal epithelial barrier function immediately after the episode but that a rapid and more severe acidification (pH 5.2) in vitro increases epithelial permeability following the insult.

Changes in Bacterial Diversity Associated with Epithelial Tissue in the Beef Cow Rumen during the Transition to a High-Grain Diet

ABSTRACT Our understanding of the ruminal epithelial tissue-associated bacterial (defined as epimural bacteria in this study) community is limited. In this study, we aimed to determine whether diet influences the diversity of the epimural bacterial community in the bovine rumen. Twenty-four beef heifers were randomly assigned to either a rapid grain adaptation (RGA) treatment (n = 18) in which the heifers were allowed to adapt from a diet containing 97% hay to a diet containing 8% hay over 29 days or to the control group (n = 6), which was fed 97% hay. Rumen papillae were collected when the heifers were fed 97%, 25%, and 8% hay diets. PCR-denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR analysis were used to characterize rumen epimural bacterial diversity and to estimate the total epimural bacterial population (copy numbers of the 16S rRNA gene). The epimural bacterial diversity from RGA heifers changed (P = 0.01) in response to the rapid dietary transition, whereas it was not affected in control heifers. A total of 88 PCR-DGGE bands were detected, and 44 were identified from phyla including Firmicutes, Bacteroidetes, and Proteobacteria. The bacteria Treponema sp., Ruminobacter sp., and Lachnospiraceae sp. were detected only when heifers were fed 25% and 8% hay diets, suggesting the presence of these bacteria is the result of adaptation to the high-grain diets. In addition, the total estimated population of rumen epimural bacteria was positively correlated with molar proportions of acetate, isobutyrate, and isovalerate, suggesting that they may play a role in volatile fatty acid metabolism in the rumen.

Evaluation of triticale dried distillers grains with solubles as a substitute for barley grain and barley silage in feedlot finishing diets

The objective of this study was to assess the value of triticale dried distillers grains with solubles (DDGS) as a replacement for barley silage in addition to a portion of the dry-rolled barley (DRB) in a grain-based feedlot finishing diet. The trial used 160 crossbred yearling steers: 144 noncannulated (478 +/- 84 kg) in a complete randomized design, and 16 ruminally cannulated (494 +/- 50 kg) in a replicated 4 x 4 Latin square design. The noncannulated steers were assigned to 8 standard pens (10 per pen) and 8 pens equipped with the GrowSafe system (GrowSafe Systems Ltd., Airdrie, Alberta, Canada; 8 per pen). The cannulated steers were placed (2 per pen) in the 8 GrowSafe pens and moved between pens at 28-d intervals. Each of 4 experimental diets was fed in 2 standard and 2 GrowSafe pens. The diets contained (DM basis) 1) 85% DRB and 10% barley silage (CON); 2) 65% DRB, 20% triticale DDGS, and 10% barley silage (D-10S), 3) 65% DRB, 25% triticale DDGS, and 5% barley silage, and 4) 65% DRB, 30% triticale DDGS, and no barley silage. Supplement (5% of dietary DM) was included in all diets. Ruminal pH was measured over four 7-d periods using indwelling electrodes. Replacing barley silage with triticale DDGS linearly decreased mean ruminal pH (P = 0.006), linearly increased duration (P = 0.006 and P = 0.01) and area under the curve (P = 0.02 and P = 0.05) below pH 5.5 and 5.2, and linearly increased the frequency of subacute (P = 0.005) and acute (P = 0.05) bouts of ruminal acidosis. Variation in mean ruminal pH decreased (P = 0.008) in steers fed D-10S compared with CON. Similarly, variation in DMI was less for steers fed triticale DDGS compared with CON. Steers fed D-10S tended to have greater DMI (P = 0.08) but similar ADG and G:F compared with CON steers. Replacing barley silage with triticale DDGS tended to linearly decrease DMI (P = 0.10) and increase (P = 0.06) G:F. Compared with CON, steers fed D-10S tended to have greater backfat thickness (P = 0.10) and decreased dressing percentage (P = 0.06), ribeye area (P = 0.10), and meat yield (P = 0.06). Severity and number of abscessed livers was greater (P = 0.006) in steers fed D-10S compared with those fed CON. Although mean ruminal pH decreased as barley silage was replaced with triticale DDGS, the trend for improved growth suggests that reduced ruminal pH did not affect animal performance. Triticale DDGS can be substituted for barley silage in finishing diets in addition to a portion of barley grain without affecting growth performance or carcass quality, but it is recommended that an antimicrobial be included in the diet to reduce liver abscesses.