G.B. Penner

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.

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.

Effects of partially replacing barley silage or barley grain with dried distillers grains with solubles on rumen fermentation and milk production of lactating dairy cows.

Dried distillers grains with solubles (DDGS) has been commonly used as a dietary protein source for lactating dairy cows. However, there is a paucity of data evaluating the use of DDGS as a partial replacement of forage or grain. The objective of this study was to determine the effects of partially replacing barley silage or barley grain with corn/wheat-based DDGS on dry matter intake (DMI), chewing activity, rumen fermentation, and milk production. Six ruminally cannulated lactating Holstein cows were used in a replicated 3 x 3 Latin square design with 21-d periods. Cows were fed the control diet (CON: 45% barley silage, 5% alfalfa hay, and 50% concentrate mix), a low forage (LF) diet or a low grain (LG) diet, in which barley silage or barley grain was replaced by DDGS at 20% of dietary dry matter, respectively. All diets were formulated to contain 18% crude protein and fed as total mixed rations. Compared with CON, cows fed the LF diet had greater DMI (26.0 vs. 22.4 kg/d), yields of milk (36.4 vs. 33.0 kg/d), milk protein (1.18 vs. 1.05 kg/d), and milk lactose (1.63 vs. 1.46 kg/d), but milk fat yield was not affected. The LF diet decreased chewing time compared with the CON diet (29.7 vs. 39.1 min/kg of DMI), but did not affect rumen pH and duration of rumen pH below 5.8. Compared with CON, feeding the LG diet tended to increase minimum and maximum rumen pH, but did not affect DMI, milk yield, and milk composition in this study. These results indicate that a partial replacement of barley silage with DDGS can improve the productivity of lactating dairy cows without negatively affecting rumen fermentation and milk fat production. Barley grain can also be partially replaced by DDGS in diets for lactating dairy cows without causing negative effects on productivity.

Effects of feeding Fermenten on ruminal fermentation in lactating Holstein cows fed two dietary sugar concentrations

This study was conducted to determine the effects of feeding Fermenten (Church and Dwight Co., Princeton, NJ) with or without dietary sucrose on ruminal fermentation, apparent total-tract nutrient digestibility, and nutrient utilization. Eight ruminally cannulated Holstein cows (163 +/- 55 d in milk; mean +/- standard deviation) were used in a replicated 4 x 4 Latin square design with a 2 x 2 factorial arrangement of treatments. Experimental diets were formulated with and without Fermenten (0 vs. 3.3% of dietary DM) at 2 dietary sugar concentrations (2.8 vs. 5.7%). Dietary treatment did not affect dry matter intake or apparent total-tract nutrient digestibility. Feeding Fermenten did not affect ruminal pH, but high-sugar diets tended to increase the daily minimum pH (5.61 vs. 5.42) and mean pH (6.17 vs. 6.30) compared with low-sugar diets. Ruminal ammonia concentration tended to be greater for cows fed Fermenten compared with control (18.1 vs. 15.9 mg/dL), but was not affected by dietary sugar concentration. Significant interactions between Fermenten and dietary sugar concentration were detected for some milk production responses. Fermenten treatment numerically increased milk fat yield (0.92 vs. 0.82 kg/d), 4% fat-corrected milk yield (24.3 vs. 21.9 kg/d), and milk energy output (18.2 vs. 16.4 Mcal/d) compared with control for cows fed low-sugar diets, but not for cows fed high-sugar diets. Increasing dietary sugar concentration did not enhance the effects of Fermenten, providing no support for the theory that synchronizing the availability of N and fermentable energy in the rumen improves nutrient utilization in lactating dairy cows.

Technical note: Evaluation of a continuous ruminal pH measurement system for use in noncannulated small ruminants

The objective of this study was to evaluate the precision and accuracy of an indwelling ruminal pH measurement system that could be used in small ruminants (small ruminant ruminal pH measurement system; SRS) without requiring ruminal cannulation. The outer diameter, length, and weight of the SRS were 20.6 mm, 138 mm, and 245 g, respectively. This device was capable of logging pH, temperature, and battery voltage. In Exp. 1, a ruminally cannulated sheep (94 kg) was infused with a 40% (wt/vol) glucose solution to supply 5 g of glucose/kg of BW into the rumen. Ruminal pH was recorded every 30 s simultaneously using a portable pH meter and the SRS. In Exp. 2, 30 noncannulated sheep (72 +/- 10 kg of BW) were orally administered with a 40% glucose solution as described above (5 g of glucose/kg of BW; n = 22) or an equivalent volume of water (12.5 mL/kg of BW; n = 8). Sheep were slaughtered 3 h after the oral drench, and immediately after slaughter ruminal pH readings were measured manually using a portable pH meter and were compared with measurements recorded by the SRS. In Exp. 1, the relationship between manual pH measurement using a portable pH meter and the SRS (226 data pairs) had a Pearson correlation coefficient and concordance correlation coefficient of 0.97 and 0.96, respectively. Furthermore, the scale shift and location shift observed in Exp. 1 were 1.28 and 0.00, respectively. The relationship between measurements conducted manually using a portable pH meter and the SRS in Exp. 2 had Pearson and concordance correlation coefficients of 0.96 and 0.95, respectively. The respective scale and location shifts for Exp. 2 were 1.16 and 0.04. These results indicate that the measurements obtained from SRS were in agreement with simultaneous measurements manually conducted using a portable pH meter, suggesting that the SRS can be used to measure ruminal pH in noncannulated small ruminants.

Effects of feeding triticale dried distillers grains plus solubles as a nitrogen source on productivity of lactating dairy cows

The objective of this study was to compare triticale dried distillers grains plus solubles (TDDGS) as a source of dietary N with other high-protein feeds commonly used in North America: corn dried distillers grains plus solubles (CDDGS), canola meal (CM), and soybean meal (SBM). Rumen degradable protein (% of crude protein, CP) after 16h of incubation in the rumen was higher for CDDGS and TDDGS (69.3% and 64.5%, respectively) than for CM (62.2%) and SBM (53.0%). For the lactation study, experimental diets were formulated to supply 30% of dietary CP from TDDGS, CDDGS, CM, or SBM. These diets contained 22.3% forage neutral detergent fiber and approximately 19.2% CP and were fed to 12 multiparous Holstein cows (130+/-40 d in milk) in a 4 x 12 Latin rectangle design with 21-d periods. Neither dry matter intake nor milk yield was affected by treatment, averaging 25.5 and 35.5kg/d, respectively. Plasma concentrations of Arg, Lys, and Thr were greater for cows fed CM or SBM compared with those fed TDDGS or CDDGS, whereas plasma concentrations of Leu and Phe were lower for cows fed CM or SBM compared with those fed TDDGS or CDDGS. Cows fed CDDGS had lower milk CP yield compared with cows fed CM (1.07 vs. 1.16kg/d). Contrarily, milk CP and milk lactose yields were not different for cows fed TDDGS compared with CM or SBM. These data suggest that TDDGS can replace CM or SBM in the diets of lactating dairy cows without adverse effects on production. Furthermore, although dried distillers grain has been generally accepted as a feed high in ruminal undegradable protein, CDDGS and TDDGS used in the present study had high in situ ruminal degradable crude protein. Further investigation is warranted to determine the extent of variation in ruminal protein degradation among different types of dried distillers grains.