Estimation of ruminal outflow in buffaloes fed diets with different energy and protein sources by use of reticular and omasal sampling.

This study aimed to evaluate the effects of different traditional or alternative energy and protein sources, associated or not, on feeding behavior, ruminal kinetics, and post-ruminal flow of nutrients. Besides, it was assessed diets' effects on different sites (reticulum and omasum) of buffaloes. Four ruminally cannulated male Murrah buffaloes (average initial weight of 637 ± 66.37 kg) were randomly distributed in a 4 × 4 Latin square design. Treatments were arranged as 2 × 2 factorial arrangement. The first factor evaluated was the inclusion of energy sources (ground corn and crude glycerin), and the second factor was the inclusion of protein sources (soybean meal and cottonseed cake). Buffaloes fed cottonseed cake had a higher content of neutral detergent fiber (NDF) and potentially digestible detergent fiber (pdNDF) in the rumen environment than buffaloes fed soybean meal. There was a sampling site effect on rumen digestion rates of pdNDF, passage rates of indigestible neutral detergent fiber (iNDF), and pdNDF, and flow of iNDF. In this study, omasal collections were more representative. Total replacement of ground corn by crude glycerin promoted less NDF ruminal digestibility, and care should be taken to include this energy source. The cottonseed cake does not cause a difference in rumen dynamics and can totally replace soybean meal in feedlot buffaloes' diet.

Rumen metabolism, omasal flow of nutrients, and microbial dynamics in lactating dairy cows fed fresh perennial ryegrass (Lolium perenne L.) not supplemented or supplemented with rolled barley grain.

The objective of this study was to evaluate the effect of rolled barley grain (RB) supplementation on rumen metabolism, omasal flow of nutrients, and microbial dynamics in lactating dairy cows fed fresh perennial ryegrass (Lolium perenne L.; PRG)-based diets. Ten ruminally cannulated Holstein cows averaging (mean ± standard deviation) 49 ± 23 d in milk and 513 ± 36 kg of body weight were assigned to 1 of 2 treatments in a switchback design. The treatment diets were PRG only (G) or PRG plus 3.5 kg of dry matter RB (G+RB). The study consisted of three 29-d periods where each period consisted of 21 d of diet adaptation and 8 d of data and sample collection. A double marker system was used to quantify nutrient flow entering the omasal canal along with labeled 15N-ammonium sulfate to measure bacterial, protozoal, and nonmicrobial N flow. Rumen evacuation techniques were used to determine nutrient and microbial pool size, allowing the calculation of fractional rates of digestion and microbial growth. There was no difference in daily milk yield or energy-corrected milk yield between treatments. Milk fat concentration and milk urea N decreased, whereas milk protein concentration increased in cows fed the G+RB diet. During the omasal sampling phase, dry matter intake was higher in cows fed the G+RB diet. Ruminal and total-tract neutral detergent fiber digestibility was lower in G+RB cows; however, no difference was observed in reticulorumen pH. The rumen pool size of fermentable carbohydrate was increased in cows fed the G+RB diet; however, the fractional rate of digestion was decreased. Flow of nonammonia N and bacterial N at the omasal canal increased in cows fed the G+RB diet compared with the G diet. Protozoa N flow was not different between diets; however, protozoa appeared to supply a much larger amount of microbial N and exhibited shorter generation time than previously considered. Feed N ruminal digestibility, corrected for microbial contribution, was similar for both treatments (88.4 and 89.0% for G and G+RB, respectively). In conclusion, RB supplementation did not benefit overall animal performance; however, it reduced ruminal neutral detergent fiber digestibility and increased bacterial N flow. The results demonstrate the large dependence of cows consuming PRG-based diets on microbial N as the main source of nonammonia N supply. Additional quantitative research is required to further describe the supply of nutrients and microbial dynamics in cows consuming PRG-based diets in an effort to determine most limiting nutrients.

Comparison of microbial fermentation data from dual-flow continuous culture system and omasal sampling technique: A meta-analytical approach.

Although the omasal sampling technique (OST) has been successfully used to estimate ruminal fermentation and nutrient flow, alternatives to invasive animal trials should be pursued and evaluated. The objective of this study was to evaluate carbohydrate and N metabolisms using a meta-analytical approach to compare 2 methods: dual-flow continuous culture system (DFCCS) and OST. To be included, studies needed to report diet chemical composition and report at least 1 of the dependent variables of interest. A total of 155 articles were included, in which 97 used the DFCCS and 58 used the OST. The independent variables used were dietary nonfiber carbohydrate concentration, neutral detergent fiber (NDF) degradability, true crude protein (CP) degradability, and efficiency of microbial protein synthesis (EMPS). In addition, 12 dependent variables were used. Statistical analyses were performed using the Mixed procedure of SAS (SAS Institute Inc., Cary, NC). A random coefficients model was used considering study as a random effect and including the possibility of covariance between the slope and the intercept. The effect of method (DFCCS or OST) was included and tested in the estimates of the intercept, linear, and quadratic effects of the independent variable. There was no method effect when NDF degradability was regressed with total volatile fatty acids concentration, true CP degradability, and EMPS. Molar proportions of acetate and propionate were quadratically associated with NDF degradability. When NDF degradability was regressed with acetate and propionate there was a method effect, differing only in the intercept (ß0) estimate. True organic matter digestibility, bacterial N/total N, efficiency of N utilization, total volatile fatty acid concentration, and molar proportion of butyrate linearly increased as dietary nonfiber carbohydrate concentration increased, and none of these variables were affected by method. Concentration of ammonia N had a linear and positive association with true CP degradability. This was the only variable that had a method effect when regressed with true CP degradability, differing only in the estimate of the intercept (ß0). As EMPS increased, efficiency of N utilization also increased, and it was affected by method. Overall, the majority of DFCCS responses were similar to OST. When a method effect was observed, it was mainly on the estimate of the intercept, demonstrating that the magnitude of these responses was different. However, the relationships between independent and dependent variables were similar across methods.

Nitrogen utilization, whole-body urea-nitrogen kinetics, omasal nutrient flow, and production performance in dairy cows fed lactose as a partial replacement for barley starch.

The objective of this study was to determine whether the partial replacement of barley starch with lactose (fed as dried whey permeate; DWP) affects N utilization, whole-body urea kinetics, and production in dairy cows. Eight lactating Holstein cows were used in a replicated 4 × 4 Latin square design with 28-d periods. Four cows in one Latin square were ruminally cannulated and used to determine dietary effects on whole-body urea kinetics and N utilization. Cows were fed a barley-based diet that contained 3.6% (dry matter basis) total sugar (TSG; designated control), or diets that contained 6.6, 9.6, or 12.6% TSG. Dietary TSG content was increased by the replacement of barley grain with DWP (83% lactose). Diets were isonitrogenous (∼17.3% crude protein), and starch contents of the control, 6.6, 9.6, and 12.6% TSG diets were 24.3, 22.2, 21.2, and 19.1%, respectively. Whole-body urea kinetics were measured using 4-d infusions of [15N15N]-urea with concurrent total collections of feces and urine. Dry matter intake (mean = 26.7 kg/d), milk yield (mean = 34.9 kg/d), and milk protein and fat contents were unaffected by diet. Ruminal ammonia-N concentration decreased linearly as TSG content increased, whereas ruminal butyrate concentration increased linearly as TSG content increased. Urinary excretion of total N and urea-N changed quadratically, whereas urinary excretion of total N (% of N intake) tended to change quadratically as TSG content increased. Fecal N excretion linearly increased as TSG content increased. A quadratic response was observed for total N excretion as TSG content increased. Milk N and retained N were not affected by diet. As TSG content increased, we observed quadratic responses in the omasal flow of fluid-associated and total bacterial nonammonia N, endogenous production of urea-N, urea-N recycled to the gastrointestinal tract, and urea-N returned to the ornithine cycle. Dietary TSG content did not affect the anabolic utilization of recycled urea-N or the proportion of recycled urea-N that was used for bacterial growth. Our results indicate that feeding DWP did not influence dry matter intake, milk yield, or milk composition. Feeding DWP decreased ruminal ammonia-N concentration, but this did not result in positive responses in milk protein secretion or N balance. The quadratic response in omasal flow of total bacterial nonammonia N indicated that including TSG beyond 9.6% of diet dry matter might depress ruminal microbial protein synthesis.

Temporal changes in milk fatty acid composition during diet-induced milk fat depression in lactating cows.

Diet-induced milk fat depression (MFD) in lactating cows has been attributed to alterations in ruminal lipid metabolism leading to the formation of specific fatty acid (FA) biohydrogenation intermediates that directly inhibit milk fat synthesis. However, the mechanisms responsible for decreased lipid synthesis in the mammary gland over time are not well defined. The aim of this study was to evaluate the effect of diet on milk FA composition and milk fat production over time, especially during MFD, and explore the associations between MFD and FA biohydrogenation intermediates in omasal digesta and milk. Four lactating Finnish Ayrshire cows used in a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments and 35-d experimental periods were fed diets formulated to cause differences in ruminal and mammary lipid metabolism. Treatments consisted of an iso-nitrogenous total mixed ration based on grass silage with a forage to concentrate ratio of 65:35 or 35:65 without added oil, or with sunflower oil at 50 g/kg of diet dry matter. The high-concentrate diet with sunflower oil (HSO) induced a 2-stage drop in milk fat synthesis that was accompanied by specific temporal changes in the milk FA composition. The MFD on HSO was associated especially with trans-10 18:1 and also with trans-9,cis-11 conjugated linoleic acid (CLA) in milk and omasal digesta across all diets and was accompanied by the appearance of trans-10,cis-15 18:2. Trans-10,cis-12 CLA was increased in HSO, but milk fat secretion was not associated with omasal or milk trans-10,cis-12 CLA. The temporal changes in milk fat content and yield and milk FA composition reflect the shift from the predominant ruminal biohydrogenation pathway to an alternative pathway. The ambiguous role of trans-10,cis-12 CLA suggests that trans-10 18:1, trans-9,cis-11 CLA and trans-10,cis-15 18:2 or additional mechanisms contributed to the diet-induced MFD in lactating cows.

Effects of a commercial fermentation byproduct or urea on milk production, rumen metabolism, and omasal flow of nutrients in lactating dairy cattle.

The objective of this study was to evaluate the effects of a fermentation byproduct on rumen fermentation and microbial yield in high producing lactating dairy cattle. Eight ruminally cannulated multiparous Holstein cows averaging (mean ± standard deviation) 60 ± 10 d in milk and 637 ± 38 kg of body weight were assigned to 1 of 2 treatment sequences in a switchback design. Treatment diets contained (dry matter basis) 44% corn silage, 13% alfalfa silage, 12% ground corn, and 31% premix containing either a control mix of urea and wheat middlings (CON) or a commercial fermentation byproduct meal (Fermenten, Arm and Hammer Animal Nutrition, Princeton, NJ) at 3% diet inclusion rate (EXP). Diets were formulated to be isonitrogenous and isocaloric, with similar levels of neutral detergent fiber and starch. The trial consisted of three 28-d experimental periods, where each period consisted of 21 d of diet adaptation and 7 d of data and sample collection. Omasal nutrient flows were determined using a triple-marker technique and double-labeled 15N15N-urea. The EXP diet provided 18 g/d more nonammonia N versus the CON diet, representing 3.0% of total N intake. Energy-corrected milk yield (41.7 and 43.1 kg/d for CON and EXP, respectively), milk fat, and protein yield and content did not differ between treatments. Total dry matter intake was similar between treatments (25.5 and 26.4 kg/d for CON and EXP, respectively). Ammonia N concentration and pool size in the rumen was greater in cows fed the EXP diet. No differences were observed in rumen or total-tract dry matter, organic matter, or neutral detergent fiber digestibility. Ruminal degradation of feed N was 15% lower in cows fed EXP diets, resulting in differences in omasal N flows. Results demonstrated the fermentation byproduct meal had a sparing effect on degradable feed protein, but did not increase microbial N flow from the rumen.

Rumen digestion kinetics, microbial yield, and omasal flows of nonmicrobial, bacterial, and protozoal amino acids in lactating dairy cattle fed fermentation by-products or urea as a soluble nitrogen source.

The objective of this study was to evaluate the effect of a fermentation by-product on rumen function, microbial yield, and composition and flows of nutrients from the rumen in high-producing lactating dairy cattle. Eight ruminally cannulated multiparous Holstein cows averaging (mean ± standard deviation) 60 ± 10 d in milk and 637 ± 38 kg of body weight were randomly assigned to 1 of 2 treatment sequences in a switchback design. Treatment diets contained (dry matter basis) 44% corn silage, 13% alfalfa silage, 12% ground corn, and 31% protein premix, containing either a control mix of urea and wheat middlings (CON) or a commercial fermentation by-product meal (Fermenten, Arm and Hammer Animal Nutrition, Princeton, NJ) at 3% diet inclusion rate (EXP). The trial consisted of three 28-d experimental periods, where each period consisted of 21 d of diet adaptation and 7 d of data and sample collection. A triple-marker technique and double-labeled 15N15N-urea were used to were used to measure protozoal, bacterial, and nonmicrobial omasal flow of AA. Rumen pool sizes and omasal flows were used to determine digestion parameters, including fractional rates of carbohydrate digestion, microbial growth, and yield of microbial biomass per gram of degraded substrate. Fermentation by-product inclusion in EXP diets increased microbial N and amino acid N content in microbes relative to microbes from CON cows fed the urea control. Microbial AA profile did not differ between diets. Daily omasal flows of AA were increased in EXP cows as a result of decreased degradation of feed protein. The inclusion of the fermentation by-product increased nonmicrobial AA flow in cows fed EXP versus CON. Average protozoal contribution to microbial N flow was 16.8%, yet protozoa accounted for 21% of the microbial AA flow, with a range of 8 to 46% for individual AA. Cows in this study maintained an average rumen pool size of 320 g of microbial N, and bacterial and protozoal pools were estimated at 4 different theoretical levels of selective protozoa retention. Fractional growth rate of all microbes was estimated to be 0.069 h-1, with a yield of 0.44 g of microbial biomass per gram of carbohydrate degraded. Results indicated that fermentation by-product can increase omasal flow of AA while maintaining adequate rumen N available for microbial growth and protein synthesis. Simulations from a developmental version of the Cornell Net Carbohydrate and Protein System indicated strong agreement between predicted and observed values, with some areas key for improvement in AA flow and bacterial versus protozoal N partitioning.

Mechanism of peptide absorption in the isolated forestomach epithelial cells of dairy cows.

BACKGROUND: Peptide absorption from the forestomach plays a vital role in protein nutrition of dairy cows. This study was conducted to investigate the mechanism of dipeptide absorption in the forestomach of dairy cows using isolated omasal epithelial cells (OECs) and ruminal epithelial cells (RECs). RESULTS: Compared with RECs, the OECs formed a less tight monolayer, but had greater ability to transport glycylsarcosine (Gly-Sar) (P < 0.05). The OEC monolayers were immunopositive for the antibodies of anti-junction proteins. Gly-Sar transport was significantly greater at 37 °C than that at 4 °C, with an optimal pH of 6.0-6.5, and was decreased significantly by diethylpyrocarbonate and dipeptide Met-Gly (P < 0.05). The apical-to-basolateral transport was significantly greater than basolateral-to-apical transport (P < 0.05). Knockdown of peptide transporter 1 (PepT1) resulted in less Gly-Sar uptake in OECs, whereas overexpression of PepT1 in OECs resulted in higher Gly-Sar uptake (P < 0.05). Additionally, the expression of PepT1 was upregulated by the treatment with various dipeptides (P < 0.05). CONCLUSION: The OECs have a greater ability to transport Gly-Sar than RECs do. Both passive and active routes are involved in the process of Gly-Sar absorption in the isolated cultured forestomach epithelial cells from dairy cows. © 2018 Society of Chemical Industry.

Effect of exogenous butyrate on the gastrointestinal tract of sheep. I. Structure and function of the rumen, omasum, and abomasum.

The aim of this study was to determine the effect of exogenous butyrate on the structure and selected functions of the stomach in sheep. Eighteen rams (30.8 ± 2.1 kg; 12 to 15 mo of age) were allocated to the study and fed a diet for 14 d without (CTRL) or with sodium butyrate (BUT; 36 g/kg of offered DM). Neither DMI nor initial BW differed between treatments (P ≥ 0.61), but final BW was greater for BUT compared with CTRL (P = 0.03). Butyrate concentration in the reticuloruminal fluid and abomasal digesta was greater for BUT compared with CTRL (P ≤ 0.01), but total short-chain fatty acids (SCFA) concentration, as well as concentration of other SCFA, did not differ between treatments (P ≥ 0.07). Relative to BW, reticuloruminal tissue mass tended (P = 0.09) to be greater and omasal digesta was less (P = 0.02) for BUT compared with CTRL. Dietary butyrate did not affect ruminal papillae length, width, and density nor did it affect ruminal epithelium thickness (P ≥ 0.12) in the ventral sac of the rumen. However, the DM of ruminal epithelium (mg/cm2) tended (P = 0.06) to be greater for BUT compared with CTRL. Omasal and abomasal epithelium thicknesses were greater (P ≤ 0.05) for BUT compared with CTRL. Mitosis-to-apoptosis ratio in the abomasal epithelium was less for BUT compared with CTRL (P = 0.04). Finally, the mRNA expression of peptide transporter 1 in the omasal epithelium was less (P = 0.02) and mRNA expression of monocarboxylate transporter 1 in the abomasal epithelium tended (P = 0.07) to be greater for BUT compared with CTRL. It can be concluded that exogenous butyrate supplementation affected not only the rumen but also omasum and abomasum in sheep.

Effects of maturity at harvest on the nutritive value and ruminal digestion of Eragrostis tef (cv. Moxie) when fed to beef cattle.

Teff (Eragrostis tef cv. Moxie), a warm-season annual grass, could be an excellent forage for beef cattle. However, there is limited information on its nutritive value to cattle when harvested at different stages of maturity. Thus, the objective of this research was to determine the effect of feeding teff hay harvested at the boot (BT), early-heading (EH), or late-heading (LH) stages of maturity on nutrient intake, ruminal fermentation characteristics, omasal nutrient flow, and N utilization in beef cattle. Six ruminally cannulated beef heifers (mean initial BW ± SD, 476 ± 32.6) were used in a replicated 3 × 3 Latin square design with 28-d periods (18 d for adaptation and 10 d for measurements). Dry matter intake was measured daily. Indwelling pH loggers were used to measure ruminal pH from days 21 to 28. Ruminal fluid and omasal digesta were collected from days 26 to 28 to determine fermentation characteristics and omasal nutrient flow. Fecal and urine samples to quantify N excretion were also collected (days 26 to 28). Blood samples for plasma urea-N (PUN) determination were collected 3 h post-feeding on day 28. There were no changes (P > 0.28) in the ADF or NDF content of teff with advancing maturity, but indigestible NDF increased (P < 0.01) with increasing maturity. Maturity had no effect (P ≥ 0.14) on DMI, and ruminal total short-chain fatty acid (SCFA) concentration, pH, digestibility, and outflow of DM, OM, NDF, ADF, and CP. However, the CP content of BT hay was greater (P < 0.01) than for EH and LH hay (18.1, 14.1, and 11.5%, respectively, DM basis), and this resulted in the higher CP intake (P < 0.01) for heifers fed the BT than the EH and LH hay. Consequently, ruminal ammonia-N (NH3-N) concentration was greater (P < 0.01) for heifers fed BT than EH and LH hay, thereby possibly explaining the tendency for a decrease (P = 0.08) in PUN concentration, and a decrease (P < 0.01) in the excretion of total N, urine N, and urea-N (UUN) with advancing maturity. However, fecal N excretion (g/d) did not differ (P = 0.76). In conclusion, despite a decrease in CP intake and ruminal NH3-N concentration, feeding beef heifers EH and LH compared to BT teff hay did not compromise ruminal digestion and outflow of DM, OM, NDF, ADF, and CP, and microbial protein synthesis. Advancing maturity in teff hay also resulted in a decrease in the excretion of total N and urine N and UUN when fed to cattle.

Absorption of bicarbonate in sheep omasum.

Transport of bicarbonate across the isolated epithelium of sheep omasum was studied in vitro in Ussing chambers in combination with the pH-Stat method. The transport of HCO3- occurred in both directions, but Jms HCO3- was significant larger than Jsm. Reducing the activity of the apical Na/H exchanger by a low mucosal Na concentration caused a significant reduction of Jms HCO3-. Mucosal amiloride or short chain fatty acids (25 mmol l-1 SCFA) numerically decreased Jms HCO3-, but their combination (amiloride + SCFA) caused a significant reduction, which was also observed after addition of the carboanhydrase inhibitor ethoxyzolamide. Concentrations of 5 or 15 mmol·l-1 mucosal ammonia did not change transport rates. The obtained results indicate the importance of an undisturbed cytosolic pH for transcellular HCO3- transport, which is probably mediated by an anion exchanger in both the apical and basolateral membranes. Possible impairment of HCO3- transport appears to be an overlooked factor in the pathogenesis of displacement of the abomasum.

Effect of dietary fish oil supplements alone or in combination with sunflower and linseed oil on ruminal lipid metabolism and bacterial populations in lactating cows.

Fish oil (FO) alters ruminal biohydrogenation causing trans fatty acid (FA) intermediates to accumulate, but the effects of 18-carbon polyunsaturated FA supply on ruminal long-chain FA metabolism and microbial communities in cattle fed FO are not well established. Four cows fitted with rumen cannula were used in a 4 × 4 Latin square with 21-d experimental periods to evaluate the effects of FO alone or in combination with plant oils high in 18:2n-6 or 18:3n-3 on rumen microbial ecology and flow of FA at the omasum. Treatments comprised a basal grass silage-based diet containing no additional oil (control) or supplements of FO (200 g/d) or FO (200 g/d) plus 500 g/d of sunflower oil (SFO) or linseed oil (LFO). Flow of FA was determined using the omasal sampling technique. The relative abundance of key biohydrogenating bacteria was assessed by quantitative PCR on 16S rRNA genes in omasal digesta. Fish oil-supplemented treatments increased the amounts of trans-18:1, trans-18:2, and 20- to 22-carbon polyunsaturated FA escaping the rumen. Relative to the control, oil supplements had no effect on the amount of 18:0 leaving the rumen, but LFO decreased the flow of 18:0 at the omasum compared with SFO. Both SFO and LFO increased trans-18:1 relative to FO, whereas LFO resulted in the highest trans-18:2 and 20- to 22-carbon FA flow. Supplements of FO plus plant oils shifted biohydrogenation toward trans-10 18:1 formation. Compared with FO alone, the ruminal metabolism of 22:6n-3 in the rumen of lactating cows is more extensive on diets containing higher amounts of 18-carbon polyunsaturated FA. However, the biohydrogenation of 22:5n-3 was less extensive in LFO than SFO, but showed no difference between FO and diets containing plant oils. Ruminal outflow of 20:5n-3 was not altered when plant oils were added to FO. Alterations in the amount of intermediates at the omasum or ruminal biohydrogenation pathways were not accompanied by major changes in analyzed bacterial populations. In conclusion, dietary supplements of FO alone or in combination with plant oils increase the amount of biohydrogenation intermediates containing 1 or more trans double bonds escaping the rumen, which may have implications for host metabolism and the nutritional quality of ruminant foods.

Effect of ruminal acidosis and short-term low feed intake on indicators of gastrointestinal barrier function in Holstein steers.

The objective of this study was to determine effect of ruminal acidosis (RA) and low feed intake [LFI] on the regional barrier function of the gastrointestinal tract. Twenty-one Holstein steers were fed for ad libitum intake for 5 d (control [CON]), fed at 25% of ad libitum intake for 5 d (LFI), or provided 2 d of ad libitum intake followed by 1-d of feed restriction (25% of ad libitum intake), 1 d where 30% of ad libitum dry matter intake (DMI) was provided as pelleted barley followed by the full allocation (RA) and fed for ad libitum intake the following day. Tissues and digesta from the rumen, omasum, duodenum, jejunum, ileum, cecum, proximal, and distal colon were collected. Permeability was assessed using the mucosal-to-serosal flux of inulin (JMS-inulin) and mannitol (JMS-mannitol). Digesta pH was 0.81, 0.63, and 0.42 pH units less for RA than CON in the rumen, cecum, and proximal colon; while, LFI had pH that was 0.47 and 0.36 pH units greater in the rumen and proximal colon compared to CON. Total ruminal short-chain fatty acid (SCFA) concentration were less for LFI (92 mM; P = 0.010) and RA (87 mM; P = 0.007) than CON (172 mM) steers. In the proximal colon, the proportion of butyrate (P = 0.025 and P = 0.022) and isobutyrate (P = 0.019 and P = 0.019) were greater, and acetate (P = 0.028 and P = 0.028) was less for LFI and RA, respectively, when compared to CON steers. Ruminal papillae length, width, perimeter, and surface area were 1.21 mm, 0.78 mm, 3.84 mm, and 11.15 mm2 less for LFI than CON; while, RA decreased papillae width by 0.52 mm relative to CON. The JMS-mannitol was less for LFI steers than CON in the proximal colon (P = 0.041) and in the distal colon (P = 0.015). Increased gene expression for claudin 1, occludin, tight-cell junction protein 1 and 2, and toll-like receptor 4 were detected for LFI relative to CON in the rumen, jejunum, and proximal colon. For RA steers, expression of toll-like receptor 4 in the rumen, and occludin and tight-cell junction protein 1 were greater in the jejunum than CON. An acute RA challenge decreased pH in the rumen and large intestine but did not increase tissue permeability due to increases in the expression of genes related to barrier function within 1 d of the challenge. Steers exposed to LFI for 5 d had reduced ruminal SCFA concentrations, smaller ruminal papillae dimensions, and increased tissue permeability in the proximal and distal colon despite increases for genes related to barrier function and immune function.

Predicting omasal flow of nonammonia N and milk protein yield from in vitro-determined utilizable crude protein at the duodenum.

This study evaluated the relationship between utilizable crude protein (uCP) at the duodenum estimated in vitro and omasal flow of crude protein (CP; omasal flow of nonammonia N × 6.25) measured in lactating dairy cows. In vivo data were obtained from previous studies estimating omasal digesta flow using a triple-marker method and 15N as microbial marker. A total of 34 different diets based on grass and red clover silages were incubated with buffered rumen fluid previously preincubated with carbohydrates for 3 h. The buffer solution was modified to contain 38 g of NaHCO3 and 1 g of (NH4)HCO3 in 1,000 mL of distilled water. Continuous sampling of the liquid phase for determination of ammonia-N was performed at 0.5, 4, 8, 12, 24, and 30 h after the start of incubation. The ammonia N concentrations after incubation were used to calculate uCP. The natural logarithm of uCP [g/kg of dry matter (DM)] at time points 0.5, 4, 8, 12, 24, and 30 h of incubation was plotted against time to estimate the concentration of uCP (g/kg of DM) at time points 16, 20, and 24 h using an exponential function. Fixed model regression analysis and mixed model regression analysis with random study effect were used to evaluate the relationships between predicted uCP (supply and concentration) and observed omasal CP flow and milk protein yield. Residual analysis was also conducted to evaluate whether any dietary factors influenced the relationships. The in vitro uCP method ranked the diets accurately in terms of total omasal CP flow (kg/d) or omasal CP flow per kilogram of DM intake. We also noted a close relationship between estimated uCP supply and adjusted omasal CP flow, as demonstrated by a coefficient of determination of 0.87, although the slope of 0.77 indicated that estimated uCP supply (kg/d) was greater than the value determined in vivo. The linear bias with mixed model analysis indicated that uCP supply overestimated the difference in omasal CP flow between the diets within a study, an error most likely related to study differences in feed intake, animals, and methodology. Predicting milk protein yield from uCP supply showed a positive relationship using a mixed model (coefficient of determination = 0.79), and we observed no difference in model fit between the time points of incubation (16, 20, or 24 h). The results of this study indicate that the in vitro method can be a useful tool in evaluating protein value of ruminant diets.

Effects of replacing soybean meal with canola meal or treated canola meal on ruminal digestion, omasal nutrient flow, and performance in lactating dairy cows.

Extrusion treated canola meal (TCM) was produced in an attempt to increase the rumen-undegraded protein fraction of canola meal (CM). The objective of this study was to evaluate the effects of replacing soybean meal (SBM) with CM or TCM on ruminal digestion, omasal nutrient flow, and performance in lactating dairy cows. To assess performance, 30 multiparous Holstein cows averaging (mean ± SD) 119 ± 23 d in milk and 44 ± 7 kg of milk/d and 15 primiparous cows averaging 121 ± 19 d in milk and 34 ± 6 kg of milk/d were blocked in a randomized complete block design with a 2-wk covariate period and 12-wk experimental period (experiment 1). Dietary ingredients differed only in protein supplements, which were SBM, CM, or TCM. All diets were formulated to contain (dry matter basis) 30% alfalfa silage, 30% corn silage, 4% soy hulls, 2.4% mineral-vitamin premix, and 16% CP. The SBM diet contained 25% high-moisture shelled corn and 8.6% SBM; the canola diets contained 22% high-moisture shelled corn and either 11.2% CM or 11.4% TCM. To assess ruminal digestion and omasal nutrient flow, 6 rumen-cannulated cows were blocked into 2 squares of 3 cows and randomly assigned within blocks to the same 3 dietary treatments as in experiment 1 in a replicated 3 × 3 Latin square design (experiment 2). Data were analyzed using the MIXED procedure of SAS (SAS Institute, Cary, NC). Orthogonal contrasts were used to compare effects of different protein supplements: SBM versus CM + TCM and CM versus TCM. In experiment 1, compared with SBM, apparent total-tract digestibilities of dry matter and nutrients were greater in cows fed both CM diets, and there was a tendency for nutrient digestibilities to be higher in cows fed CM compared with TCM. Diets did not affect milk yield and milk components; however, both canola diets decreased urinary urea N (% of total urinary N), fecal N (% of total N intake), and milk urea N concentration. In experiment 2, compared with SBM, both canola diets increased N intake and tended to increase rumen-degraded protein supply (kg/d) and N truly digested in the rumen (kg/d). Diets did not affect ruminal digestibility, efficiency of microbial protein synthesis, and rumen-undegraded protein flow among diets. Results from this experiment indicate that replacing SBM with CM or TCM in diets of lactating cows improved digestibility and may reduce environmental impact. Moreover, under the conditions of the present study, treating CM by extrusion did not improve CM utilization.

Effect of organic grass-clover silage on fiber digestion in dairy cows.

There are differences in grass-clover proportions and chemical composition between herbage from primary growth (PG) and regrowth (RG) in grass-clover leys. Mixing silages made from PG and RG may provide a more optimal diet to dairy cows than when fed separately. We tested the hypotheses that increasing dietary proportions of grass-clover silage made from RG compared with PG would increase digestion rate of potentially degradable NDF (pdNDF), and increase ruminal accumulation of indigestible NDF (iNDF). Eight rumen cannulated Norwegian Red cows were used in two replicated 4×4 Latin squares with 21-day periods. Silages were prepared from PG and RG of an organically cultivated ley, where PG and RG silages were fed ad libitum in treatments with RG replacing PG in ratios of 0, 0.33, 0.67 and 1 on dry matter basis in addition to 8 kg concentrate. We evaluated the effect of the four diets with emphasis on rumen- and total tract fiber digestibility. Increasing RG proportions decreased silage intake by 7%. Omasal flow of pdNDF decreased, whereas iNDF flow increased with increasing RG proportions. Increasing RG proportions decreased rumen pool sizes of NDF and pdNDF, whereas pool sizes of iNDF and CP increased. Increasing RG proportions increased digestion rate of NDF, which resulted in greater total tract digestion of NDF. Pure PG diet had the highest calculated energy intake, but the improved rumen digestion of NDF by cows offered 0.33 and 0.67 of RG leveled out milk fat and protein yields among the three PG containing diets.

Acidic pH and short-chain fatty acids activate Na+ transport but differentially modulate expression of Na+/H+ exchanger isoforms 1, 2, and 3 in omasal epithelium.

Low sodium content in feed and large amounts of salivary sodium secretion are essential requirements to efficient sodium reabsorption in the dairy cow. It is already known that Na(+)/H(+) exchange (NHE) of the ruminal epithelium plays a key role in Na(+) absorption, and its function is influenced by the presence of short-chain fatty acids (SCFA) and mucosal pH. By contrast, the functional role and regulation of NHE in omasal epithelium have not been completely understood. In the present study, we used model studies in small ruminants (sheep and goats) to investigate NHE-mediated Na(+) transport and the effects of pH and SCFA on NHE activity in omasal epithelium and on the expression of NHE isoform in omasal epithelial cells. Conventional Ussing chamber technique, primary cell culture, quantitative PCR, and Western blot were used. In native omasal epithelium of sheep, the Na(+) transport was electroneutral, and it was inhibited by the specific NHE3 inhibitor 3-[2-(3-guanidino-2-methyl-3-oxo-propenyl)-5-methyl-phenyl]-N-isopropylidene-2-methyl-acrylamide dihydrochloride, which decreased mucosal-to-serosal, serosal-to-mucosal, and net flux rates of Na(+) by 80% each. The application of low mucosal pH (6.4 or 5.8) in the presence of SCFA activated the Na(+) transport across omasal epithelium of sheep compared with that at pH 7.4. In cultured omasal epithelial cells of goats, mRNA and protein of NHE1, NHE2, and NHE3 were detected. The application of SCFA increased NHE1 mRNA and protein expression, which was most prominent when the culture medium pH decreased from 7.4 to 6.8. At variance, the mRNA and protein expression of NHE2 and NHE3 were decreased with low pH and SCFA, which was contrary to the published data from ruminal epithelial studies. In conclusion, this paper shows that (1) NHE1, NHE2, and NHE3 are expressed in omasal epithelium; (2) NHE3 mediates the major portion of transepithelial Na(+) transport in omasal epithelium; and (3) SCFA and acidic pH acutely activate Na(+) transport but suppress the expression of NHE2 and NHE3 in the longer term. By contrast, the expression of NHE1 is increased by SCFA and acidic pH, indicating a prominent role for NHE1 in the regulation of intracellular pH of omasal epithelium. Our results suggest a regulatable Na(+) absorption in ruminal and omasal epithelium. It is of benefit for intracellular pH homeostasis and highly relevant to dairy cows fed on high-concentrate diets.

Effects of feeding canola meal or wheat dried distillers grains with solubles as a major protein source in low- or high-crude protein diets on ruminal fermentation, omasal flow, and production in cows.

The objective of this study was to determine the effects of feeding canola meal (CM) or wheat dried distillers grains with solubles (W-DDGS) as the major source of protein in diets varying in crude protein (CP) content on ruminal fermentation, microbial protein production, omasal nutrient flow, and production performance in lactating dairy cows. Eight lactating dairy cows were used in a replicated 4×4 Latin square design with 29-d periods (21 d of dietary adaptation and 8 d of measurements) and a 2×2 factorial arrangement of dietary treatments. Four cows in 1 Latin square were ruminally cannulated to allow ruminal and omasal sampling. The treatment factors were (1) source of supplemental protein (CM vs. W-DDGS) and (2) dietary CP content (15 vs. 17%; DM basis). Diets contained 50% forage and 50% concentrate, and were fed twice daily at 0900 and 1600 h as total mixed rations for ad libitum intake. Dry matter intake and milk yield were unaffected by dietary treatments; however, milk yield in cows that were fed CM was numerically greater (+1.1 kg/d) when compared with cows fed W-DDGS. Feeding CM increased milk lactose content compared with feeding W-DDGS. Milk urea nitrogen and ruminal NH3-N concentrations were greater in cows fed the high-CP compared with those fed the low-CP diet. The rumen-degradable protein supply was greater in cows fed the high-CP when compared with those fed the low-CP diet when diets contained CM, whereas rumen-degradable protein supply was lower in cows fed the high-CP when compared with those fed the low-CP diet when diets contained W-DDGS. Total N flow at the omasal canal was not affected by diet; however, omasal flow of NH3-N was greater in cows fed CM when compared with those fed W-DDGS. The rumen-undegradable protein supply was greater in cows fed the low-CP when compared with those fed the high-CP diet when diets contained CM, whereas rumen-undegradable protein supply was lower in cows fed the low-CP when compared with those fed the high-CP diet when diets contained W-DDGS. Omasal flow of fluid-associated bacteria was greater and that of particle-associated bacteria tended to be greater in cows fed CM when compared with those fed W-DDGS; however, omasal flow of total microbial nonammonia N was unaffected by dietary treatment. Omasal flows of threonine and tryptophan were greater, whereas that of histidine and lysine tended to be greater in cows fed CM when compared with those fed W-DDGS. Our results show that when dairy diets are formulated to contain 15 or 17% CP, CM or W-DDGS can be used as the major source of protein and achieve similar levels of milk production.

The influence of age and weaning on permeability of the gastrointestinal tract in Holstein bull calves.

Fourteen Holstein bull calves were used in a randomized complete block design to investigate the effect of calf age and weaning on permeability of the gastrointestinal tract (GIT). Calves were randomly assigned to 1 of 2 treatments: (1) a weaning protocol that was initiated on d 35; WN; n=7), or (2) a control treatment where calves were not weaned (CON; n=7). Calves were bottle-fed milk replacer (150 g/L), in 3 equal portions/d targeting 15% of their body weight (BW) in liquid milk intake [approximately 21.1g/kg of BW/d, dry matter (DM) basis]. On d 35, the amount of milk replacer offered to WN calves was reduced to 7.5% of BW for 7 d before calves were weaned on d 42. On d 14, 28, and 42, calves were orally dosed with 500 mL of Cr-EDTA (179 mM Cr-EDTA solution) and housed in a metabolism crate to enable total urine collection and determination of total urinary Cr recovery as an indicator of total-tract permeability. On d 44, calves were killed and tissues from the rumen, omasum, duodenum, jejunum, ileum, cecum, and proximal and distal colon were collected, rinsed, and transported in buffer solution (pH 7.4 at 38.5°C). Tissues were incubated in Ussing chambers under short-circuit conditions with buffer solutions designed to mimic the mucosal and serosal energy source that would be available in vivo (glucose for tissues from the small intestine and short-chain fatty acids for tissues that would be exposed to fermentation; rumen, omasum, and large intestinal tissues). The serosal to mucosal flux of (14)C-mannitol and (3)H-inulin was measured for each region. Although we detected treatment × period interactions for BW and starter intake, dietary treatments did not differ within a week. Overall, the time that ruminal pH was <5.5 was less before weaning than after weaning. We observed a differential response for the appearance of Cr in urine for WN and CON calves, where the appearance of Cr (mg/48 h) in urine decreased for both treatments from d 14 to 28, but increased from d 28 to 42 for WN, whereas Cr appearance continued to decrease for CON. The flux of mannitol and inulin did not differ between treatments but did differ among region of the GIT, with rumen, duodenum, and jejunum having the greatest permeability. These data suggest that permeability of the GIT decreases with age but weaning may disrupt this process. The rumen, duodenum, and jejunum appear to be the regions with greatest permeability.

Parameterization of a ruminant model of phosphorus digestion and metabolism.

The objective of the current work was to parameterize the digestive elements of the model of Hill et al. (2008) using data collected from animals that were ruminally, duodenally, and ileally cannulated, thereby providing a better understanding of the digestion and metabolism of P fractions in growing and lactating cattle. The model of Hill et al. (2008) was fitted and evaluated for adequacy using the data from 6 animal studies. We hypothesized that sufficient data would be available to estimate P digestion and metabolism parameters and that these parameters would be sufficient to derive P bioavailabilities of a range of feed ingredients. Inputs to the model were dry matter intake; total feed P concentration (fPtFd); phytate (Pp), organic (Po), and inorganic (Pi) P as fractions of total P (fPpPt, fPoPt, fPiPt); microbial growth; amount of Pi and Pp infused into the omasum or ileum; milk yield; and BW. The available data were sufficient to derive all model parameters of interest. The final model predicted that given 75 g/d of total P input, the total-tract digestibility of P was 40.8%, Pp digestibility in the rumen was 92.4%, and in the total-tract was 94.7%. Blood P recycling to the rumen was a major source of Pi flow into the small intestine, and the primary route of excretion. A large proportion of Pi flowing to the small intestine was absorbed; however, additional Pi was absorbed from the large intestine (3.15%). Absorption of Pi from the small intestine was regulated, and given the large flux of salivary P recycling, the effective fractional small intestine absorption of available P derived from the diet was 41.6% at requirements. Milk synthesis used 16% of total absorbed P, and less than 1% was excreted in urine. The resulting model could be used to derive P bioavailabilities of commonly used feedstuffs in cattle production.