Sensitivity analysis of the INRA 2018 feeding system for ruminants by a one-at-a-time approach: Effects of dietary input variables on predictions of multiple responses of dairy cattle.

In the INRA 2018 feeding system for ruminants, the prediction of multiple animal responses is based on the integration of the characteristics of the animal and the available feedstuff characteristics, as well as the rationing objectives. In this framework, the characterization of feedstuffs in terms of net energy, digestible protein, and fill units requirs information on their chemical composition, digestibility, and degradability. Despite the importance of these feed characteristics, a comprehensive assessment of their impact on the responses predicted by the INRA 2018 feeding system has not been carried out. Thus, our study investigated how variables predicted by the INRA feeding system (i.e., outputs) for dairy cows are affected by variation in feed characterization (i.e., inputs). Five input variables were selected for the sensitivity analysis (SA): CP, OM apparent digestibility (OMd), GE, effective degradability of nitrogen assuming a passage rate of 6%/h (ED6_N) and true intestinal digestibility (dr_N) of nitrogen. A one-at-a-time SA was performed on predicted digestive, productive and environmental output variables for dairy cows with 6 contrasted diets. These 6 diets were formulated to meet 95% of the potential daily milk production (37.5 kg) of a multiparous cow at wk 14 of lactation. Then, the values of the 5 key input variables of each feedstuff were randomly sampled around the INRA 2018 feed table values (reference point). The response of the output variable to the variation of the input variable was quantified and compared using the tangent value at the reference point and the normalized sensitivity coefficient. Among the major final output variables, CP and dr_N had the greatest impact on nitrogen (N) excretion in urine (as a proportion of total fecal and urinary N excretion, UN/TN), OMd and GE had the greatest impact on N utilization efficiency (N in milk as proportion of intake N, NUE), and ED6_N had the greatest impact on milk protein yield (MPY). Additionally, CP, GE, and dr_N had the least effect on methane emission, OMd had the least effect on UN/TN, and ED6_N had the least effect on NUE. The responses of most output variables to ED6_N and dr_N variations were highly dependent on diet, and were related to the ratio between PDI (i.e., metabolizable protein) and UFL (i.e., NEL) at the reference point of each diet. In conclusion, we were able to analyze the response of output variables to the variations of the input variables, using the tangent and its normalized value at the reference point. The predicted final outputs were more impacted by variations in CP, GE, and OMd. The other 2 input variables, ED6_N and dr_N, had a smaller effect on the final output variables, but the responses varied between the diets according to their PDI/UFL ratio. Among the final output variables affected by ED6_N, MPY was the most impacted, but when quantified this impact was at an acceptable level. Our present study was conducted using 6 representative diets for dairy cattle fed at their potential, but should be completed by the analysis of more diverse conditions.

Exploring metabolism flexibility in complex organisms through quantitative study of precursor sets for system outputs.

BACKGROUND: When studying metabolism at the organ level, a major challenge is to understand the matter exchanges between the input and output components of the system. For example, in nutrition, biochemical models have been developed to study the metabolism of the mammary gland in relation to the synthesis of milk components. These models were designed to account for the quantitative constraints observed on inputs and outputs of the system. In these models, a compatible flux distribution is first selected. Alternatively, an infinite family of compatible set of flux rates may have to be studied when the constraints raised by observations are insufficient to identify a single flux distribution. The precursors of output nutrients are traced back with analyses similar to the computation of yield rates. However, the computation of the quantitative contributions of precursors may lack precision, mainly because some precursors are involved in the composition of several nutrients and because some metabolites are cycled in loops. RESULTS: We formally modeled the quantitative allocation of input nutrients among the branches of the metabolic network (AIO). It corresponds to yield information which, if standardized across all the outputs of the system, allows a precise quantitative understanding of their precursors. By solving nonlinear optimization problems, we introduced a method to study the variability of AIO coefficients when parsing the space of flux distributions that are compatible with both model stoichiometry and experimental data. Applied to a model of the metabolism of the mammary gland, our method made it possible to distinguish the effects of different nutritional treatments, although it cannot be proved that the mammary gland optimizes a specific linear combination of flux variables, including those based on energy. Altogether, our study indicated that the mammary gland possesses considerable metabolic flexibility. CONCLUSION: Our method enables to study the variability of a metabolic network with respect to efficiency (i.e. yield rates). It allows a quantitative comparison of the respective contributions of precursors to the production of a set of nutrients by a metabolic network, regardless of the choice of the flux distribution within the different branches of the network.

[13C] GC-C-IRMS analysis of methylboronic acid derivatives of glucose from liver glycogen after the ingestion of [13C] labeled tracers in rats.

We developed a complete method to measure low [(13)C] enrichments in glycogen. Fourteen rats were fed a control diet. Six of them also ingested either [U-(13)C] glucose (n=2) or a mixture of 20 [U-(13)C] amino acids (n=4). Hepatic glycogen was extracted, digested to glucose and purified on anion-cation exchange resins. After the optimization of methylboronic acid derivatization using GC-MS, [(13)C] enrichment of extracted glucose was measured by GC-C-IRMS. The accuracy was addressed by measuring the enrichment excess of a calibration curve, which observed values were in good agreement with the expected values (R=0.9979). Corrected delta values were -15.6+/-1.6 delta(13)C (per thousand) for control rats (n=8) and increased to -5 to 8 delta(13)C (per thousand) per thousand and 12-14 delta(13)C (per thousand) per thousand after the ingestion of [U-(13)C] amino acids or [U-(13)C] glucose as oral tracers, respectively. The method enabled the determination of dietary substrate transfer into glycogen. The sequestration of dietary glucose in liver glycogen 4 h after the meal was 35% of the ingested dose whereas the transfer of carbon skeletons from amino acids was only 0.25 to 1%.

Ingestion of a protein hydrolysate is accompanied by an accelerated in vivo digestion and absorption rate when compared with its intact protein.

BACKGROUND: It has been suggested that a protein hydrolysate, as opposed to its intact protein, is more easily digested and absorbed from the gut, which results in greater plasma amino acid availability and a greater muscle protein synthetic response. OBJECTIVE: We aimed to compare dietary protein digestion and absorption kinetics and the subsequent muscle protein synthetic response to the ingestion of a single bolus of protein hydrolysate compared with its intact protein in vivo in humans. DESIGN: Ten elderly men (mean +/- SEM age: 64 +/- 1 y) were randomly assigned to a crossover experiment that involved 2 treatments in which the subjects consumed a 35-g bolus of specifically produced L-[1-(13)C]phenylalanine-labeled intact casein (CAS) or hydrolyzed casein (CASH). Blood and muscle-tissue samples were collected to assess the appearance rate of dietary protein-derived phenylalanine in the circulation and subsequent muscle protein fractional synthetic rate over a 6-h postprandial period. RESULTS: The mean (+/-SEM) exogenous phenylalanine appearance rate was 27 +/- 6% higher after ingestion of CASH than after ingestion of CAS (P < 0.001). Splanchnic extraction was significantly lower in CASH compared with CAS treatment (P < 0.01). Plasma amino acid concentrations increased to a greater extent (25-50%) after the ingestion of CASH than after the ingestion of CAS (P < 0.01). Muscle protein synthesis rates averaged 0.054 +/- 0.004% and 0.068 +/- 0.006%/h in the CAS and CASH treatments, respectively (P = 0.10). CONCLUSIONS: Ingestion of a protein hydrolysate, as opposed to its intact protein, accelerates protein digestion and absorption from the gut, augments postprandial amino acid availability, and tends to increase the incorporation rate of dietary amino acids into skeletal muscle protein.

Changes in mammary uptake and metabolic fate of glucose with once-daily milking and feed restriction in dairy cows.

The aim of this review is to better understand the regulation of milk yield in response to once-daily milking and feed restriction. Glucose is the principal precursor for the synthesis of lactose (a major osmotic agent in milk), and participates in determining the milk volume produced. When applying these two breeding factors, reductions in milk yield are associated with a reduction in milk lactose yield and in the arterial flow of glucose, due to a decrease in the mammary blood flow. The ability of the udder to extract glucose is altered with once-daily milking but not necessarily with feed restriction. Lactose synthesis is down-regulated in response to once-daily milking and feed restriction but the percentage of the extracted glucose which is converted into lactose is differently affected in response to treatments. No marked change is observed with once daily milking whereas this would be increased with feed restriction and in contrast, depressed with fasting.

Effect of site of starch digestion on portal nutrient net fluxes in steers.

Processing of maize grain is known to modulate the site of starch digestion, thus the nature and amount of nutrients delivered for absorption. We assessed the effect of site of starch digestion on nutrient net fluxes across portal-drained viscera (PDV). Three steers, fitted with permanent digestive cannulas and blood catheters, successively received two diets containing 35 % starch as dent maize grain. Diets differed according to maize presentation: dry and cracked (by-pass, BP) v. wet and ground (control, C). Ruminal physicochemical parameters were not significantly affected. Between C and BP, the decrease in ruminal starch digestion was compensated by an increase in starch digestion in the small intestine. The amount of glucose and soluble alpha-glucoside reaching the ileum was not affected. The amount of glucose disappearing in the small intestine increased from 238 to 531 g/d between C and BP, but portal net flux of glucose remained unchanged (-97 g/d). The portal O2 consumption and net energy release were not significantly affected, averaging 16 % and 57 % of metabolizable energy intake, respectively. The whole-body glucose appearance rate, measured by jugular infusion of [6,6-2H2]glucose, averaged 916 g/d. The present study shows that the increase in the amount of glucose disappearing in the small intestine of conventionally fed cattle at a moderate intake level induces no change in portal net flux of glucose, reflecting an increase in glucose utilization by PDV. That could contribute to the low response of whole-body glucose appearance rate observed at this moderate level of intestinal glucose supply.

Validation of the measurement of glucose appearance rate with [6,6-2H2]glucose in lactating dairy cows.

The aim of this study was to validate the measurement of glucose appearance rate using [6,6-2H2]glucose i.v. infusion in lactating dairy cows. Sample enrichments were analysed by gas chromatography/mass spectrometry. Linearity (enriched solutions) and specificity (enriched plasma) were good: for enrichments ranging between 1.6 and 6.3 mol% excess, the slopes were about 1 and the ordinates at the origin were not different from zero. For a plasma enriched at 3.74 mol% excess, repeatability and long term intralaboratory reproducibility coefficients of variation were 1.31 and 1.90%, respectively. The appearance rates were calculated by two models. The values provided by the steady-state model were not different from those provided by the non-steady-state Steele model. Both models can be used because the treatment effects were similarly discriminated regardless of the model. In our experiments analysing the nutritional effects on Ra in mid-lactating cows, the precision of the method (1.90%) was not the limiting factor to detect a significant difference in Ra compared to the statistical precision obtained with the experimental scheme (4 x 4 and 5 x 5 Latin square design). We conclude that in lactating dairy cows, the measurement of glucose fluxes with this method is relevant and minimally invasive for the animals.

Short-term mild hyperglycemia enhances insulin-stimulated glucose disposal in lactating goats.

This work was designed to study the effect of a 3-day mild hyperglycemia (5.3 vs. 3.3 mM) on the regulation of glucose metabolism in lactating goats. Glucose was intravenously infused at variable rates simultaneously with a constant potassium-amino acid infusion. Diet plus substrate infusion maintained net energy but not protein supply. Milk yield did not change. Skeletal muscle glucose transporter (GLUT-4) was analyzed before and after hyperglycemia. In addition, the acute effect of medium and high insulin doses on glucose turnover was measured in vivo during euglycemic and hyperglycemic hyperinsulinemic clamps under potassium and amino acid replacement. Hyperglycemia reduced the endogenous glucose appearance but increased glucose disposal. It decreased the total membrane-associated GLUT-4 protein in skeletal muscle. In contrast, it improved the acute insulin-stimulated glucose disposal. Both the level and duration (3 days) of hyperglycemia contributed to this improvement. We conclude that short-term mild hyperglycemia has similar effects in lactating goats as those already observed in nonlactating rodents or humans.