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The aim of this study was to determine whether subtle changes in the energy-to-N ratio of medium-concentrate diets alters hepatic export protein synthesis and the partition of protein metabolism in the whole body of growing rams. Rams (n = 6; 41.5 [+ or -] 2.6 kg of BW) were fitted with catheters for measurement of Leu and Phe tracer kinetics across the portal drained-viscera (PDV) and liver. Rams were assigned to receive 3 dietary treatments according to a duplicated Latin square design. Animals received forage-concentrate-based diets that were balanced for ME and available N (CON), 20% imbalanced (reduced) in available N (LN), or 20% imbalanced in ME (LE). After 15 d on each experimental diet, [ring-[sup.2][H.sub.5]]Phe (4.3 [micro]mol x [kg.sup.-l] x [h.sup.-1]) and [1-[sup.13]C]Leu (8.6 [micro]mol x [kg.sup.-l] x [h.sup.-1]) were continuously infused into the vena cava for 10 h and, over the last 7 h of infusion, matched sets of blood samples were taken. Daily BW gain was less (P < 0.05) for the LE (0.191 kg x [d.sup.-1]) diet compared with CON (0.265 kg/d) and LN (258 kg x [d.sup.-1]) diets. Compared with CON, whole body irreversible loss rate (ILR) of Leu and Phe was less (10 to 16%, P < 0.02) for LN and LE diets, which for Leu reflected its decreased (20 to 24%, P < 0.05) net PDV absorption. The decreased whole body ILR is due to a decreased PDV ILR in both diets with a relative contribution of the PDV to the whole body ILR decreased (P < 0.05) in the LN (27%) diet compared with the CON (36%) and LE (33%) diets. This decreased PDV ILR was associated with a decreased net Leu PDV uptake in LN and LE diets (-25 and -20%, respectively; P < 0.05). Conversely, the decreased whole body Phe ILR is explained by a decreased hepatic ILR (and contribution to the whole body ILR) and was associated with a decreased net hepatic uptake of Phe in LN (-25%) and LE (-20%) diets compared with CON (P = 0.03). The fractional and absolute synthesis rates of total proteins and albumin were decreased by 10% in LE animals (P < 0.05), whereas they were not affected by the LN diet. These results suggest a specific decreased utilization of Leu at the PDV due to a specific sparing mechanism in the LN diet. Conversely, a decreased Phe utilization occurred in the liver in both diets (due to a decreased export protein synthesis and a probable decreased oxidation in LE diet, whereas only oxidation is reduced in LN diet). Key words: amino acid, energy, liver, metabolic flux, ruminant, stable isotope doi: 10.2527/jas.2009-2062

The objective of this study was to evaluate the usefulness of correcting net portal absorption (NPA) of urea-N, ammonia, and AA-N for differences in methods before their inclusion into a meta-analysis. It was hypothesized that the difference, or portal-drained viscera (PDV) balance, between N inputs (apparently digested N plus urea-N) and outputs (ammonia plus AA-N) was 0 in the absence of measurement errors and based on the assumption that other sources of N inputs and outputs were relatively small and balanced each other. A database was built from 44 publications that reported data from 129 treatments (sheep, n = 71; beef cattle, n = 32; and dairy cows, n = 26). When necessary, NPA of urea-N (n = 38) and ammonia (n = 35) results were recalculated on a whole-blood basis, whereas NPA of AA-N (n = 87) was recalculated for all the N from AA transferred across the PDV rather than only the N from the [alpha]-amino group. Before corrections, PDV balance averaged 22.9% of N ingested (SD 29.0) for all treatments; after corrections, PDV balance significantly decreased to 10.2% of N ingested (SD 34.7). No difference in PDV balance was observed among species before or after corrections. Correcting NPA of urea-N, ammonia, and AA-N increased the accuracy without improving precision. Therefore, from a biological perspective, recalculating reported data seems appropriate to reduce bias due to differences in methods because this approach reduces the excess in N inputs relative to N outputs. Key words: amino acid, ammonia, meta-analysis, portal flux, ruminant, urea

The aim of this study was to assess the metabolic fate of AA (endogenous or export protein synthesis, gluconeogenesis, or oxidation) after an imbalanced supply of energy and N in the diet of growing lambs. Eighteen INRA 401 lambs (3 mo old, 29.7 [+ or -] 0.45 kg of BW) were fed 3 experimental diets, one providing a N and energy supply according to recommended allowances (control), one with 23% less N supply relative to energy (LN), and one with 19% less ME supply relative to N (LE). Animals were assigned to 6 blocks of 3, with each animal receiving 1 of the 3 diets, and the animals from each block were slaughtered on the same day. Liver slices from these lambs were incubated in a minimum salt medium (Krebs-Henseleit) containing physiological concentrations of propionate and AA as energy and N sources, similarly across all 3 treatments. Protein synthesis (endogenous and export) using [U-[sup.14]C]valine and [[sup.35]S]methionine, gluconeogenesis from [1-[sup.14]C]propionate and [U-[sup.14]C]alanine, and oxidation were measured. A relative sparing of AA at the liver level was observed with the LN diet because of reduced urinary N (-42%, LN vs. control, P < 0.001). The AA were also directed toward anabolic purposes in the LN diet via an increased endogenous and total export protein synthesis ([+ or -]51%, LN vs. control, P = 0.01; also observed for fibrinogen synthesis, but not for albumin or transferrin) associated with a tendency for increased gluconeogenesis from alanine ([+ or -]58%, LN vs. LE, P = 0.08). On the other hand, the LE diet resulted in a marked loss of N in urine ([+ or -]24%, LE vs. control, P < 0.05), but no notable effect of the LE diet was demonstrated for protein synthesis or gluconeogenesis ex vivo. These data demonstrate a more efficient utilization of AA for anabolic purposes in the lambs fed LN, probably via an activation of some AA transport systems, to address the shortage of nitrogenous nutrients in the LN diet. By contrast, no such adaptation occurred in the LE lambs, probably because the regulatory mechanisms that prevailed in this case were the nutrient supply or hormones, which were not altered in our ex vivo experimental model. Key words: amino acid, ex vivo, gluconeogenesis, lamb, liver, protein synthesis

The current trend in energy feeding systems for ruminants toward a nutrient-based system requires dietary energy supply to be determined in terms of amount and nature of absorbed energy-yielding nutrients. The objective of this study was to establish response equations on the net portal appearance (NPA) of VFA and glucose, and their secondary metabolites [beta]-hydroxybutyrate (BHBA) and lactate, to changes in intake level and chemical dietary characteristics based on the Institut National de la Recherche Agronomique Feed Evaluation System for Ruminants. Meta-analyses were applied on published data compiled from the FLORA database, which pools the results on net splanchnic nutrient fluxes in multi-catheterized ruminants from international publications. For each nutrient, several prediction variables were tested. We obtained robust models for intakes up to 30 g of DM x [d.sup.-1] x kg of [BW.sup.-1] and diets containing less than 70 g of concentrate per 100 g of DM. These models were designed to predict the NPA (mmol x [h.sup.-1] x kg of [BW.sup.-1]) of total VFA based on the amount of ruminally fermented OM (RfOM) intake [adjusted [R.sup.2] ([R.sup.2.sub.adj]) = 0.95; residual means square errors (RMSE) = 0.24], to predict VFA profile (mol/100 mol of total VFA) based on type of RfOM intake (acetate: [R.sup.2.sub.adj] = 0.85, RMSE = 2.2; propionate: [R.sup.2.sub.adj] = 0.76, RMSE = 2.2; butyrate: [R.sup.2.sub.adj] = 0.76, RMSE = 1.09), and to predict the NPA (mmol x [h.sup.-1] x kg of [BW.sup.-1]) of glucose based on the starch digested in the small intestine independent of ruminant species, and while presenting no interfering factors on the residuals and individual slopes. The model predicting the NPA (mmol x [h.sup.-1] x kg of [BW.sup.-1]) of BHBA based on the amount of RfOM intake ([R.sup.2.sub.adj] = 0.91; RMSE = 0.036) was species-dependent, and the model predicting NPA (mmol x [h.sup.-1] x kg of [BW.sup.-1]) of lactate based on starch digested in the rumen ([R.sup.2.sub.adj] = 0.77; RMSE = 0.042) presented a wide dispersion. However, the NPA (mmol x [h.sup.-1] x kg of [BW.sup.-1]) of BHBA was related to the NPA of both butyrate ([R.sup.2.sub.adj] = 0.85; RMSE = 0.054) and acetate ([R.sup.2.sub.adj] = 0.85; RMSE = 0.052), and the NPA (mmol x [h.sup.-1] x kg of [BW.sup.-1]) of lactate was related to the NPA of propionate ([R.sup.2.sub.adj] = 0.51; RMSE = 0.096). This research showed that it is possible to accurately predict the amount and nature of absorbed nutrient fluxes based on dietary characteristics in both sheep and cattle. This work aims to quantify the consequences of digestion and portal-drained viscera metabolism on nutrient availability. These results can provide deeper insight into biological processes and help develop improved tools for dietary formulation. Key words: diet composition, energy, intake level, meta-analyses, portal-drained viscera, ruminant

Splanchnic metabolism of nitrogenous nutrients and their uptake by the hind limb were studied in finishing lambs receiving ryegrass harvested at grazing stage with or without barley supplementation. Six multicatheterized lambs (40.2 [+ or -] 1.5 kg) were fed with frozen ryegrass (RG) at 690 kJ of ME intake (MEI) * [d.sup.-1] * B[W.sup.-0.75] and 20.8 g of N intake (NI)/d successively without and with barley supplementation (RG + B), according to a crossover design. Barley supplementation represented 21% of DM intake and increased the MEI and the NI by 32 and 24% respectively, (P < 0.01). In the ruminal fluid, barley increased acetate and butyrate concentrations by 21.2 and 49.6%, respectively (P < 0.04), without any effect on the ammonia concentration. Consequently, the net portal appearance (NPA) of ammonia was not modified, but the NPA of total amino acids (TAA; +38%) and nonessential amino acids (NEAA; +45%) was increased (P < 0.05) by barley supplementation. Taken individually, the NPA of the essential amino acids (EAA) was increased for isoleucine (+32%; P < 0.05), threonine (+151%; P < 0.03), and lysine (+26%; P < 0.06), with no effect for the other EAA. In contrast to what was observed at the PDV level, no significant alteration in the net hepatic amino acid flux was observed for TAA, EAA, NEAA, branched-chain amino acids (BCAA), urea, and ammonia after barley supplementation, showing a relatively minor role of the liver in the regulation of the supply of amino acids to the peripheral tissues. However, taken individually, the net hepatic uptake of some NEAA involved in gluconeogenesis and/or ureagenesis was altered with barley supplementation: the alanine uptake was increased by 44% (P < 0.05), aspartate + asparagine (asx) uptake was decreased by 18% (P < 0.01), and glutamate + glutamine (glx) release tended (P < 0.10) to be increased by 208%. With barley supplementation, NI increased by 5 g of N/d, and net splanchnic release increased by 4.63 g of N/d. Consequently, the additional dietary N supply (together with energy supply) was nearly exclusively available to peripheral tissues as AA-N (N as amino acids), but no strong effect of this additional supply of AA to the hind limb could be demonstrated in terms of net AA hind limb fluxes. Consequently, barley supplementation of a ryegrass-based diet increased the net AA release by the splanchnic tissues, with little effect on the AA net uptake by the peripheral tissues. Key Words: Amino Acids, Barley, Hind Limb, Liver, Lolium, Portal Vein

Splanchnic metabolism of energy-yielding nutrients and their uptake by the hind limb were studied in finishing lambs receiving ryegrass harvested at grazing stage (ear at 10 cm) with or without barley supplementation. Six ruminally cannulated and multi-catherized lambs (40.2 [+ or -] 1.5 kg) were fed with frozen ryegrass (RG) at 690 kJ of metabolizable energy intake (MEI) * [d.sup.-1] * B[W.sup.-0.75] successively with and without barley supplementation (RG + B), according to a triplicated Latin square design. Barley supplementation represented 21% of DM intake and increased the MEI by 32% (P < 0.002). In ruminal fluid, barley supplementation increased the acetate and butyrate concentrations by 21.2 and 49.6%, respectively (P < 0.04), without modifying those of propionate. Thus, molar proportions of acetate and butyrate were not modified, and those of propionate tended (P < 0.06) to decrease from 26 to 23%. As a result, the net portal appearance of propionate was not modified. Net portal appearance of butyrate and [beta]-hydroxybutyrate increased (P < 0.03), and that of acetate was not modified. Consequently, hepatic up take of butyrate increased and probably spared acetate from hepatic metabolism. The hepatic fractional extraction of propionate decreased (P < 0.03), whereas the net flux of lactate switched from a net release to a net uptake, suggesting an alteration in the contribution of gluconeogenic substrates to glucose synthesis without modification in net hepatic glucose release. As a consequence, barley supplementation increased net splanchnic release of acetate (P < 0.02), propionate (P < 0.001), and [beta]-hydroxybutyrate (P < 0.01) by 60, 157, and 78%, respectively. In addition, the net splanchnic release of insulin increased (P < 0.03) because of a decrease (P < 0.02) in its hepatic extraction. Despite those changes, the net uptake of nutrients by the hind limb was not modified and even decreased in the case of glucose (P < 0.02), suggesting a stimulation of lipogenesis in adipose tissues. Results from the present study suggested that supplementation of a ryegrass-based diet would likely have little effect on the orientation of muscle energy metabolism and on meat quality because the net uptake of nutrients by the hind limb was unchanged. Key Words: Barley, Energy Intake, Liver, Lolium, Nutrients, Portal Vein

Our objective was to validate the use of an ultrasonic transit-time flowmeter for the measurement of portal blood flow (PBF) in sheep. Data recorded with this technique were compared with those obtained with an indicator dilution method, and probes were calibrated in vivo. Wethers were fitted with catheters in the portal, jejunal, and ruminal veins and in a mesenteric artery. Ultrasonic flow probes were implanted around the portal vein: S-series probes in three wethers, and A-series probes in four wethers. The PBF measured with A-series probes was within 10% of that measured by indicator dilution, but PBF measured with S-series probes were 52 to 77% of that determined by indicator dilution. In vivo calibration indicated that A-series probes provided accurate measurement of PBF (absolute accuracy: 5% [+ or -] zero flow error). In conclusion, an ultrasonic transit-time flowmeter, with an A-series probe, can be used to reliably measure PBF in sheep. Key Words: Sheep, Blood Flow, Portal Vein

Improving the ability of animals to convert feed resources into food for humans is needed for more sustainable livestock systems. Genetic selection for animals eating less while maintaining their performance (i.e., low residual feed intake [RFI]) appears a smart strategy but its effectiveness relies on high-throughput animal phenotyping. Here, we explored plasma nitrogen (N) isotope ratios in an attempt to identify easily superior young bulls in terms of RFI. For this, 48 Charolais young bulls fed two contrasting diets (corn vs. grass silage diets) were selected from a larger population as extreme RFI animals (24 low-RFI vs. 24 high-RFI) and their plasma analyzed for natural 15N abundance (δ15N) in the whole protein (bulk protein) and in the individual protein-bound amino acids (PbAA). For the first time, we showed that the δ 15N in plasma bulk protein differed (P = 0.007) between efficient (low-RFI) and inefficient (high-RFI) cattle regardless of diet. Furthermore, most analyzed PbAA followed the same trend as the bulk protein, with lower (P < 0.05) δ 15N values in more efficient (low-RFI) compared with less efficient (high-RFI) cattle, again regardless of diet. The only three exceptions were Phe, Met, and Lys (P > 0.05) for which the first metabolic reaction before being catabolized does not involve transamination, a pathway known naturally to enrich AAs in 15N. The contrasted isotopic signatures across RFI groups only in those PbAA undergoing transamination are interpreted as differences in transamination rates and N-use efficiency between low- and high-RFI phenotypes. Natural isotopic N signatures in bulk proteins and specific PbAA can be proposed as biomarkers of RFI in growing beef cattle fed different diets. However, the current study cannot delineate whether this effect only occurs post-absorption or to some extent also in the rumen. Our data support the conclusion that most efficient cattle in terms of RFI upregulate N conservation mechanisms compared with less efficient cattle and justify future research on this topic., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science.)

The aim of this study was to compare the responses in fattening performance and meat composition for high-concentrate diets rich in either starch and lipids (especially omega-3 fatty acids) or fibrous by-products. A total of 140 Charolais bulls (initially 319 ± 27 kg BW) were allocated to 3 high-concentrate diets and were fattened for up to 18 mo. The diet treatments included concentrate mixtures rich in either fiber (FR; n = 56) or starch plus linseed (diets SL and SLR; n = 56 and n = 28, respectively) and barley straw. The concentrate mix was offered ad libitum in SL and FR diets but was kept isoenergetic to the FR diet in the SLR diet. Bulls were weighed every 15 d. Feed intake was measured daily. Carcass composition was assessed for all animals slaughtered at 699 ± 65 kg BW. Meat nutritional quality traits (e.g., fat content and fatty acid composition focusing on n-6 and n-3 polyunsaturated fatty acids) were measured on the longissimus thoracis, rectus abdominis, and semitendinosus muscles. Metabolic enzyme activity (phosphofructokinase, lactate dehydrogenase, and cytochrome-c oxidase) was measured on these muscles and on liver. The SL diet bulls had greater fattening performance, BW gain (P = 0.006), and efficiency for growth (P = 0.025) at an energy intake similar to that of FR diet bulls. They also had heavier carcasses with a greater proportion of fat. However, liver samples showed no difference in specific metabolic activity. Compared to bulls fed the SL diet, bulls fed SLR consumed 15% less energy and had lower BW gain (P < 0.001) but were slightly more efficient for growth (P = 0.010). They had lower carcass weight but a greater muscle-to-fat ratio. Compared to bulls fed the FR diet, SLR bulls had lower than planned NEg intake and lower BW gain but did not have differences in body composition. Compared to the FR diet, the SL diet led to a greater omega-3 fatty acid content because of a greater supply of dietary linoleic acid, especially in lean muscle.

Para-aminohippuric acid (pAH) is a marker frequently used to measure plasma or blood flow. In sheep studies, it is recognized that its determination must include a deacetylation step to counteract the hepatic acetylation of pAH. Such a procedure is not of common usage in cattle studies although a recent suggestion of hepatic pAH acetylation in dairy cows may have important consequences for hepatic nutrient fluxes. The aims of this study were to evaluate pAH analytical methods according to international procedures of metrology and confirm hepatic acetylation of pAH in dairy cows. The effect of the matrix used to prepare the standard curve was tested, and the influence of the pAH analytical method on blood flows and subsequent net hepatic fluxes of nutrients was determined. For the first objective, accuracy profiles were established and bias, indicators of precision, and limits of quantification were reported for 2 analytical methods (without and with a pAH deacetylation step) using 2 different standard matrices (water and plasma). Second, the net hepatic flux of different nutrients was determined including or not the deacetylation step and preparing the standard curves in plasma using samples collected from 5 multicatheterized lactating Jersey cows. The choice of the matrix had a significant impact on plasma pAH concentrations as illustrated by accuracy profiles. Water matrix decreased (P < 0.01) the slope, y-intercept, and the absorbance at concentration 0 mg pAH/L of the standard curve in both methods (without and with the deacetylation), supporting that standards prepared in water should not be used to analyze plasma samples. Samples collected on cows confirmed hepatic acetylation of pAH across the liver. Deacetylation, performed using plasma as the standard matrix, increased (P < 0.05) plasma pAH concentrations from 18.4, 26, and 23.5 to 21.4, 28.9, and 27.3 mg/L in the artery, portal vein, and hepatic vein, respectively. Deacetylation decreased the hepatic venous and arterial plasma and blood flows (P < 0.05) by 9 and 55%, respectively, modifying the net hepatic flux of acetate, total amino acid, and oxygen by more than 19% (P < 0.05). In conclusion, our results highlight the importance of including a deacetylation step in the pAH analysis method in cattle studies and of using plasma as standard matrix.

Our objective was to validate the use of an ultrasonic transit-time flowmeter for the measurement of portal blood flow (PBF) in sheep. Data recorded with this technique were compared with those obtained with an indicator dilution method, and probes were calibrated in vivo. Wethers were fitted with catheters in the portal, jejunal, and ruminal veins and in a mesenteric artery. Ultrasonic flow probes were implanted around the portal vein: S-series probes in three wethers, and A-series probes in four wethers. The PBF measured with A-series probes was within 10% of that measured by indicator dilution, but PBF measured with S-series probes were 52 to 77% of that determined by indicator dilution. In vivo calibration indicated that A-series probes provided accurate measurement of PBF (absolute accuracy: 5%+/-zero flow error). In conclusion, an ultrasonic transit-time flowmeter, with an A-series probe, can be used to reliably measure PBF in sheep.