Predictions of ruminal outflow of essential amino acids in dairy cattle

The objective of this work was to update and evaluate predictions of essential AA (EAA) outflows from the rumen. The model was constructed based on previously derived equations for rumen-undegradable (RUP), microbial (MiCP), and endogenous (EndCP) protein outflows from the rumen, and revised estimates of ingredient composition and EAA composition of the protein fractions. Corrections were adopted to account for incomplete recovery of EAA during 24-h acid hydrolysis. The predicted ruminal protein and EAA outflows were evaluated against a data set of observed values from the literature. Initial evaluations indicated a minor mean bias for non-ammonia, non-microbial nitrogen flow ([RUP + EndCP]/6.25) of 16 g of N per day. Root mean squared errors (RMSE) of EAA predictions ranged from 26.8 to 40.6% of observed mean values. Concordance correlation coefficients (CCC) of EAA predictions ranged from 0.34 to 0.55. Except for Leu, all ruminal EAA outflows were overpredicted by 3.0 to 32 g/d. In addition, small but significant slope biases were present for Arg [2.2% mean squared error (MSE)] and Lys (3.2% MSE). The overpredictions may suggest that the mean recovery of AA from acid hydrolysis across laboratories was less than estimates encompassed in the recovery factors. To test this hypothesis, several regression approaches were undertaken to identify potential causes of the bias. These included regressions of (1) residual errors for predicted EAA flows on each of the 3 protein-driven EA flows, (2) observed EAA flows on each protein-driven EAA flow, including an intercept, (3) observed EAA flows on the protein-driven EAA flows, excluding an intercept term, and (4) observed EAA flows on RUP and MiCP. However, these equations were deemed unsatisfactory for bias adjustment, as they generated biologically unfeasible predictions for some entities. Future work should focus on identifying the cause of the observed prediction bias.     

An evaluation of Molly cow model predictions of ruminal metabolism and nutrient digestion for dairy and beef diets

Model evaluation, as a critical process of model advancement, is necessary to identify adequacy and consistency of model predictions. The objectives of this study were (1) to evaluate the accuracy of Molly cow model predictions of ruminal metabolism and nutrient digestion when simulating dairy and beef cattle diets; and (2) to identify deficiencies in representations of the biology that could be used to direct further model improvements. A total of 229 studies (n = 938 treatments) including dairy and beef cattle data, published from 1972 through 2016, were collected from the literature. Root mean squared errors (RMSE) and concordance correlation coefficients (CCC) were calculated to assess model accuracy and precision. Ruminal pH was very poorly represented in the model with a RMSE of 4.6% and a CCC of 0.0. Although volatile fatty acid concentrations had negligible mean (2.5% of mean squared error) and slope (6.8% of mean squared error) bias, the CCC was 0.28, implying that further modifications with respect to volatile fatty acid production and absorption are required to improve model precision. The RMSE was greater than 50% for ruminal ammonia and blood urea-N concentrations with high proportions of error as slope bias, indicating that mechanisms driving ruminal urea N recycling are not properly simulated in the model. Only slight mean and slope bias were exhibited for ruminal outflow of neutral detergent fiber, starch, lipid, total N, and nonammonia N, and for fecal output of protein, neutral detergent fiber, lipid, and starch, indicating the mechanisms encoded in the model relative to ruminal and total-tract nutrient digestion are properly represented. All variables related to ruminal metabolism and nutrient digestion were more precisely predicted for dairy cattle than for beef cattle. This difference in precision was mostly related to the model's inability to simulate low forage diets included in the beef studies. Overall, ruminal pH was poorly simulated and contributed to problems in ruminal nutrient degradation and volatile fatty acid production predictions. Residual analyses suggested ruminal ammonia concentrations need to be considered in the ruminal pH equation, and therefore the inaccuracies in predicting ruminal urea N recycling must also be addressed. These modifications to model structure will likely improve model performance across a wider array of dietary inputs and cattle type.     

A model of net amino acid absorption and utilization by the portal-drained viscera of the lactating dairy cow

A more complete understanding of amino acid (AA) metabolism by the various tissues of the body is required to improve upon current systems for predicting the use of absorbed AA. The objective of this work was to construct and parameterize a model of net removal of AA by the portal-drained viscera (PDV). Six cows were prepared with arterial, portal, and hepatic catheters and infused abomasally with 0, 200, 400, or 600 g of casein daily. Casein infusion increased milk yield quadratically and tended to increase milk protein yield quadratically. Arterial concentrations of a number of essential AA increased linearly with respect to infusion amount. When infused casein was assumed to have a true digestion coefficient of 0.95, the minimum likely true digestion coefficient for noninfused duodenal protein was found to be 0.80. Net PDV use of AA appeared to be linearly related to total supply (arterial plus absorption), and extraction percentages ranged from 0.5 to 7.25% for essential AA. Prediction errors for portal vein AA concentrations ranged from 4 to 9% of the observed mean concentrations. Removal of AA by PDV represented approximately 33% of total postabsorptive catabolic use, including use during absorption but excluding use for milk protein synthesis, and was apparently adequate to support endogenous N losses in feces of 18.4 g/d. As 69% of this use was from arterial blood, increased PDV catabolism of AA in part represents increased absorption of AA in excess of amounts required by other body tissues. Based on the present model, increased anabolic use of AA in the mammary and other tissues would reduce the catabolic use of AA by the PDV.     

Effect of dietary nitrogen content and intravenous urea infusion on ruminal and portal-drained visceral extraction of arterial urea in lactating Holstein cows

Urea extraction across ruminal and portal-drained visceral (PDV) tissues were investigated using 9 rumen-cannulated and multi-catheterized lactating dairy cows adapted to low-N (12.9% crude protein) and high-N (17.1% crude protein) diets in a crossover design. The interaction between adaptation to dietary treatments and blood plasma concentrations of urea was studied by dividing samplings into a 2.5-h period without urea infusion followed by a 2.5-h period with primed continuous intravenous infusion of urea (0.493 ± 0.012 mmol/kg of BW per h). Cows were sampled at 66 ± 14 and 68 ± 12 d in milk and produced 42 ± 1 and 36 ± 1 kg of milk/d with the high-N and low-N diets, respectively. The arterial blood urea concentration before urea infusion was 1.37 and 4.09 ± 0.18 mmol/L with low-N and high-N, respectively. Dietary treatment did not affect the urea infusion-induced increase in arterial urea concentration (1.91 ± 0.13 mmol/L). Arterial urea extraction across the PDV and rumen increased from 2.7 to 5.4 ± 0.5% and from 7.1 to 23.8 ± 2.1% when cows were changed from high-N to low-N, respectively. Urea infusion did not decrease urea extractions, implying that urea transport rates were proportional to arterial urea concentrations. Urea extraction increased more across the rumen wall than across the total PDV for low-N compared with high-N, which implies that a larger proportion of total PDV uptake of arterial urea is directed toward the rumen with decreasing N intake. The ruminal vein - arterial (RA) concentration difference for ammonia increased instantly (first sampling 15 min after initiation of infusion) to the primed intravenous infusion when cows were adapted to the low-N diet. The RA difference for ammonia correlated poorly to the ventral ruminal concentration of ammonia (r = 0.55). Relating the RA difference for ammonia to a function of both ruminal ammonia concentration and the RA difference for urea markedly improved the fit (r = 0.85), indicating that a large fraction of ammonia released to the ruminal vein is absorbed from an epithelial ammonia pool not in equilibrium with the ventral ruminal ammonia pool. Changing cows from high-N to low-N affected the relative blood urea clearance by kidneys and PDV. The clearance by the kidneys decreased from 41 to 27 ± 2 L/h and the clearance by the PDV increased from 52 to 105 ± 12 L/h when the diet was changed from high-N to low-N. In conclusion, urea transport across gut epithelia in cattle is adapting to N status and driven by mass action. Data are commensurable with a model for urea transport across gut epithelia based on regulated expression or activity of facilitative urea transporters.     

Using artificial neural networks to predict pH,ammonia, and volatile fatty acid concentrations in the rumen

The objectives of this study were (1) to predict ruminal pH and ruminal ammonia and volatile fatty acid (VFA) concentrations by developing artificial neural networks (ANN) using dietary nutrient compositions, dry matter intake, and body weight as input variables; and (2) to compare accuracy and precision of ANN model predictions with that of a multiple linear regression model (MLR). Data were collected from 229 published papers with 938 treatment means. The data set was randomly split into a training data set containing 70% of the observations and a test data set with the remaining observations. A series of ANN with a range of 1 to 9 artificial neurons in 1 hidden layer were examined, and the best one was selected to compare with the best-fitted MLR model. The performance of model predictions was evaluated by root mean square errors (RMSE) and concordance correlation coefficients (CCC) using cross-evaluations with 100 iterations. When using the ANN to predict ruminal pH and concentrations of ammonia, total VFA, acetate, propionate, and butyrate, the RMSE were 4.2, 41.4, 20.9, 22.3, 32.9, and 29.7% of observed means, respectively. The RMSE for the MLR were 4.2, 37.8, 18.3, 19.9, 29.8, and 26.6% of the observed means. The CCC for ruminal pH, ruminal concentrations of ammonia, total VFA, acetate, propionate, and butyrate were 0.57, 0.49, 0.45, 0.40, 0.52, and 0.40, using the ANN, and 0.37, 0.48, 0.40, 0.29, 0.43, and 0.35, using the MLR. Evaluations of the MLR and the ANN indicated that these 2 model forms exhibited similar prediction errors, with 4.2, 39.6, 19.6, 21.1, 31.3, and 28.1% of observed means for pH, ammonia, total VFA, acetate, propionate, and butyrate. Although the ANN increased the precision of predictions related to ruminal metabolism, it failed to improve the accuracy compared with the linear regression model.     

Development and evaluation of equations in the Cornell Net Carbohydrate and Protein System to predict nitrogen excretion in lactating dairy cows

Nitrogen excretion is of particular concern on dairy farms, not only because of its effects on water quality, but also because of the subsequent release of gases such as ammonia to the atmosphere. To manage N excretion, accurate estimates of urinary N (UN) and fecal N (FN) are needed. On commercial farms, directly measuring UN and FN is impractical, meaning that quantification must be based on predictions rather than measured data. The purpose of this study was to use a statistical approach to develop equations and evaluate the Cornell Net Carbohydrate and Protein System's (CNCPS) ability to predict N excretion in lactating dairy cows, and to compare CNCPS predictions to other equations in the literature. Urinary N was over-predicted by the CNCPS due to inconsistencies in N accounting within the model that partitioned more N to feces than urine, although predicted total N excretion was reasonable. Data to refine model predictions were compiled from published studies (n=32) that reported total collection N balance results. Considerable care was taken to ensure the data included in the development data set (n=104) accounted for >90% of the N intake (NI). Unaccounted N for the compiled data set was 1.47±4.60% (mean ± SD). The results showed that FN predictions could be improved by using a modification of a previously published equation: FN (g/d) = [[NI (g/kg of organic matter) × (1 - 0.842)] + 4.3 × organic matter intake (kg/d)] × 1.20, which, when evaluated against the compiled N balance data, had a squared coefficient of determination based on a mean study effect R(MP)(2) of 0.73, concurrent correlation coefficient (CCC) of 0.83 and a root mean square error (RMSE) of 10.38 g/d. Urinary N is calculated in the CNCPS as the difference between NI and other N excretion and losses. Incorporating the more accurate FN prediction into the current CNCPS framework and correcting an internal calculation error considerably improved UN predictions (RMSE=12.73 g/d, R(MP)(2)=0.86, CCC=0.90). The changes to FN and UN translated into an improved prediction of total manure N excretion (RMSE=12.42 g/d, R(MP)(2)=0.96, CCC=0.97) and allows nutritionists and farm advisors to evaluate these factors during the ration formulation process.     

Changes in net portal nutrient flux in response to weaning transition and ionophore supplementation in dairy calves

Dairy calf weaning is associated with ketone concentrations that exceed the levels occurring in adults, and weaning represents a potential energy loss that may be mitigated by ionophore supplementation. To assess the effects of weaning and ionophore supplementation on net nutrient flux across portal-drained viscera (PDV) tissues in dairy calves, concentrations of glucose, acetoacetate (ACAC), β-hydroxybutyrate (BHBA), nonesterified fatty acids, volatile fatty acids, lactate, pyruvate, insulin, and glucagon and PDV flux rates were determined in Jersey bull calves (n = 19) at 35, 56, 84, and 112 d of age. Calves were randomly assigned at birth to either a commercial pelleted starter without (CON) or with lasalocid (TRT; 83 mg/kg of dry matter). Calves were fed only milk replacer from d 3 to 34 (d 3 to 20 = 454 g/d; d 21 to 34 = 56 g/d). After blood sampling on d 35, calves received replacer (d 35 to 41 = 45 g/d; d 42 to 48 = 22 g/d) and had free access to the CON or TRT starter, and from d 49 to 112 they received CON or TRT ad libitum. Catheters were implanted in the portal vein and in the mesenteric vein and artery between d 21 and 28. Blood flow was measured by continuous infusion of p-aminohippurate into the mesenteric vein. Six serial samples were taken at 30-min intervals from the arterial and portal vein catheters simultaneously. Portal blood flow increased with age but did not differ between CON and TRT calves. Glucose was released preweaning and was extracted postweaning by PDV, but was not affected by ionophore. The portal flux of nonesterified fatty acids was not different from zero during any of the 4 sample ages. Fluxes of ACAC and BHBA in CON and TRT calves went from no measurable flux preweaning to a postweaning PDV release that peaked at d 84, but the d-84 release of ACAC and BHBA was lower in TRT calves. The portal flux of volatile fatty acids increased with age, and PDV release of both butyrate and propionate was lower at d 84 in TRT than in CON calves. However, TRT calves had a greater PDV release of lactate on d 84, partially compensating for the lower release of propionate. Glucagon was greater in CON than in TRT calves at d 84 and could be a response to the elevated ketogenesis observed in CON calves during this period. Changes in the metabolic profile and nutrient flux of transition calves were demonstrated in response to weaning and ionophore supplementation. Inclusion of an ionophore appeared to moderate alimentary output at a postweaning period (d 84) at which ketone concentrations have the potential to exceed the whole animal capacity for utilization.     

Validated empirical models describing the combined effect of water activity and pH on the heat resistance of spores of a psychrotolerant Bacillus cereus strain in broth and béchamel sauce

The major objective of this study was to evaluate and model the combined effect of the water activity (a(w)) and pH of the heating menstrum on the heat resistance of spores of a psychrotolerant Bacillus cereus strain isolated from béchamel sauce. Two models, a quadratic polynomial equation and a reparameterized function, were assessed for their ability to describe the combined influence of a(w) and pH on the D(85°C)-values of the B. cereus isolate in tryptone soy broth. The performance of the models was validated by challenging the models with data independently collected in broth and béchamel sauce. Both models were found to adequately describe the validation data obtained in broth. However, it was determined that in béchamel sauce the predictions of the polynomial function not only showed bias (bias factor = 1.156) but were also fail-dangerous, as they deviated from the validation data by 17.2%. The reparameterized function was determined to be a good predictor of the D(85°C)-values in béchamel sauce as it showed no bias (bias factor = 1.033) and its predictions differed by only 7.9% from the validation data. The reparameterized function can be used to provide estimates of the minimum processing conditions required to achieve desired levels of spore inactivation within the a(w) and pH ranges studied and to determine the potential changes in heat resistance of B. cereus spores when a(w) and pH are changed, for example, during product reformulation. As validation of heat resistance models is rarely performed, let alone in actual food products, the models evaluated and validated in this study (in particular the reparameterized function) are of immediate relevance to the food industry.     

Net flux of nutrients across splanchnic tissues of lactating dairy cows as influenced by dietary supplements of biotin and vitamin B12

Biotin and vitamin B₁₂ are coenzymes in reactions that are essential to propionate metabolism in dairy cows. The objective of the present studies was to determine whether an increased dietary supply of these vitamins would change the net flux of nutrients through the rumen, the portal-drained viscera (PDV), the total splanchnic tissues (TSP), and the liver. Four lactating cows equipped with ultrasonic flow probes around the right ruminal artery and the portal vein and catheters in the right ruminal vein, the portal vein, one hepatic vein, and one mesenteric artery were fed 12 times per day a mixed ration at 95% of ad libitum dry matter intake. Daily supplements of 500 mg of vitamin B₁₂ + 20 mg of biotin or no vitamin supplement (study 1) or 500 mg of vitamin B₁₂ alone or with 20 mg of biotin (study 2) were fed according to a crossover design with two 4-wk periods in each study. On the last day of each period, blood flow was recorded and blood samples were collected every 30 min for 4 h. In study 1, biotin and vitamin B₁₂ given together increased milk production and milk protein yields compared with the control diet. The supplement increased appearance of the 2 vitamins across the PDV and TSP. It also reduced the net portal appearance of ammonia and total volatile fatty acids across the PDV. In study 2, compared with the 2 vitamins together, vitamin B₁₂ alone increased glucose flux across PDV and TSP as well as its arterial concentration and PDV flux of ammonia. With the diet used in the present experiment, the major effects of the vitamin supplements seem to be mediated through changes in ruminal fermentation and gastrointestinal tract metabolism rather than by effects on hepatic metabolism.     

Mammary gland metabolite utilization in response to exogenous glucose or long-chain fatty acids at low and high metabolizable protein levels

We investigated mammary gland metabolism in lactating dairy cattle in response to energy from glucogenic (glucose; GG) or lipogenic (palm olein; LG) substrates at low (LMP) and high (HMP) metabolizable protein levels. According to a 6 × 6 Latin square design, 6 rumen-fistulated second-lactation Holstein-Friesian dairy cows (97 ± 13 d in milk) were abomasally infused with saline (LMP-C); isoenergetic infusions (digestible energy basis) of 1,319 g/d glucose (LMP-GG), 676 g/d palm olein (LMP-LG), or 844 g/d essential AA (EAA; HMP-C); or isoenergetic infusions of 1,319 g/d glucose + 844 g/d EAA (HMP-GG) or 676 g/d palm olein + 844 g/d EAA (HMP-LG). Each experimental period consisted of 5 d of continuous infusion followed by 2 d of rest. A total mixed ration (42% corn silage, 31% grass silage, and 27% concentrate on a dry matter basis) formulated to meet 100 and 83% of net energy and metabolizable protein requirements, respectively, was fed at 90% of ad libitum intake by individual cow. Arterial and venous blood samples were collected on d 5 of each period. Infusing GG or LG at the HMP level did not affect milk yield or composition differently than at the LMP level. Neither GG nor LG infusion stimulated milk protein or lactose yield, but fat yield tended to decrease with GG and tended to increase with LG. Infusion of GG increased arterial plasma concentrations of glucose and insulin and decreased concentrations of β-hydroxybutyrate (BHB), nonesterified fatty acids, long-chain fatty acids (LCFA), total AA, EAA, and group 2 AA. Infusion of LG increased arterial triacylglycerides (TAG) and LCFA but did not affect EAA concentrations. Compared with the LMP level, the HMP level increased arterial concentrations of BHB, urea, and all EAA groups and decreased the concentration of total non-EAA. Mammary plasma flow increased with GG and was not affected by LG or protein level. Uptake and clearance of total EAA and group 2 AA were affected or tended to be affected by GG × AA interactions, with their uptakes being lower and their clearances higher with GG, but only at the LMP level. Infusion of LG did not affect uptake or clearance of any AA group. The HMP level increased uptake and decreased clearance of all EAA groups and decreased non-EAA uptake. Infusion of GG tended to increase mammary glucose uptake, and tended to decrease BHB uptake only at the LMP level. Infusion of LG increased mammary uptake of TAG and LCFA and increased or tended to increase clearance of TAG and LCFA. We suspect GG increased mammary plasma flow to maintain intramammary energy and AA balance and stimulated lipogenesis in adipose, accounting for depressed arterial BHB and group 2 AA concentrations. Mammary glucose uptake did not cover estimated requirements for lactose and fat synthesis at the HMP level, except during HMP-GG infusion. Results of this study illustrate flexibility in mammary metabolite utilization when absorptive supply of glucogenic, lipogenic, and aminogenic substrate is increased.     

Effect of time duration of ruminal urea infusions on ruminal ammonia concentrations and portal-drained visceral extraction of arterial urea-N in lactating Holstein cows

The effects of a 6 versus 24h ruminal urea infusion in lactating dairy cows fed a basal diet deficient in N on ruminal ammonia concentration, arterial urea-N concentration, net portal-drained viscera (PDV) urea-N flux, arterial urea-N extraction across the PDV, and renal urea-N kinetics were investigated. Three Danish Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in major splanchnic blood vessels were randomly allocated to a 3 × 3 Latin square design with 21-d periods. Treatments were ventral ruminal infusion of water for 24h (water INF), 24-h infusion of 15 g of urea/kg of dry matter intake (DMI; 24-h INF), and 6-h infusion of 15 g of urea/kg of DMI (6-h INF). The 6-h INF was initiated 0.5h after the afternoon feeding, and ran until 2230 h. Eight sample sets of arterial, portal, and hepatic blood, ruminal fluid, and urine were obtained at 0.5h before the morning feeding and 0.5, 1.5, 2.5, 3.5, 4.5, 5.5, and 6.5h after feeding (i.e., 9 to 15.5h after the 6h infusion was terminated). A substantial decrease in DMI for 6-h INF compared with 24-h INF and water INF was observed, and it has to be recognized that DMI may have confounding effects. However, the experimental setting plan was met (i.e., to cause changes in the daily pattern of ruminal ammonia and blood urea-N concentrations). The arterial urea-N concentration for 24-h INF and 6-h INF were greater than the arterial urea-N concentration with water INF throughout the sampling window. However, the arterial urea-N concentration for 6-h INF decreased steadily with sampling time reflecting a carryover effect from the ruminal urea infusion. The ruminal ammonia concentration and net portal flux of ammonia for 6-h INF were not different from water INF; hence, no carryover effect on ruminal ammonia concentration was observed. The portal flux of urea-N was not affected by treatment (i.e., even the combination of low ruminal ammonia and high arterial urea-N concentration with 6-h INF was not used by the cow to increase the uptake of urea-N across the PDV). Arterial urea-N extraction across the PDV was increased with water INF especially from 0.5 to 3.5h postprandial relative to the urea infusion treatments, reflecting increased epithelial permeability for urea-N. This indicates that daily ruminal peak of ammonia or blood urea-N concentrations overruled potential signals from low ruminal ammonia concentration observed during the sampling window. In conclusion, dairy cows appear unable to increase transport of urea-N from blood to gut in periods with low ruminal ammonia concentrations, even in a situation with infrequent N supply and apparent carryover effects on blood urea-N. It is speculated that mechanisms responsible for downregulation of epithelial urea-N transport based on daily maximum concentrations of ammonia in the rumen or urea-N in the blood suppresses any short-term signal from low ruminal ammonia during periods with low ruminal N supply.     

Representing interconversions among volatile fatty acids in the Molly cow model

The Molly cow model uses fixed stoichiometric coefficients for predicting volatile fatty acid (VFA) production from the fermented individual dietary nutrient fractions of forage and concentrate. We previously showed that predictions of VFA production had large errors and hypothesized that it was due to a lack of representation of carbon exchange among VFA. The objectives of the present study were to add VFA interconversion equations based on thermodynamics to the Molly cow model and evaluate the effect of these additions on model accuracy and precision of VFA predictions. Previously described thermodynamic equations were introduced to represent interconversions among VFA. The model was further modified to predict de novo acetate, propionate, and butyrate production coefficients based on forage-to-concentrate ratios rather than discrete, fixed sets of coefficients for forage-based, concentrate-based, and mixed diets. Both the original model and the modified one were reparameterized and evaluated against a common data set containing 8 studies reporting pH, VFA concentration, and VFA production rates using isotope dilution techniques and 62 studies reporting VFA concentrations and pH. Evaluations after parameter estimation revealed that predictions of VFA production rates were not improved, with root mean squared prediction errors (RMSPE) of 77, 60, and 51% for acetate, propionate, and butyrate, respectively, for the revised model versus 75, 63, and 55, respectively, for the original model. The RMSPE for predictions of VFA concentrations were reduced from 28, 46, and 40% to 22, 31, and 26% for acetate, propionate, and butyrate, respectively, simply by rederiving the VFA coefficients, but minimal further improvement was achieved with the addition of thermodynamically driven interconversion equations (RMSPE of 21, 32, and 27% for acetate, propionate, and butyrate, respectively). Thus, the results indicate that thermodynamically driven interchanges among VFA, as represented in this study, may not be a primary determinant for the accuracy of predictions of net production rates. Including the effect of pH on VFA absorption reduced the mean bias of propionate production and slope bias of acetate production, but not the overall RMSPE. The larger prediction errors for VFA production as compared with concentrations suggest the data quality may not be high, or that our representation of VFA production and absorption as well as ruminal digestion is inadequate. Additional data are required to discriminate among these hypotheses.     

Effect of abomasal glucose infusion on splanchnic amino acid metabolism in periparturient dairy cows

Six Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in the portal vein, hepatic vein, mesenteric vein, and an artery were used to study the effects of abomasal glucose infusion on splanchnic AA metabolism. The experimental design was a split plot, with cow as the whole plot, treatment as the whole-plot factor and days in milk (DIM) as the subplot factor. Cows were assigned to 1 of 2 treatments: control or infusion of 1,500 g/d of glucose into the abomasum from the day of calving to 29 DIM. Cows were sampled prepartum and at 4, 15, and 29 DIM. Postpartum dry matter intake increased at a lower rate with infusion compared with the control. Arterial concentrations of all essential AA (EAA) were lower with infusion compared with the control. Net portal fluxes of His, Ile, Leu, Lys, Met, Phe, Thr, Val, Ala, Pro, Ser, and Tyr were lower with infusion compared with the control and the net portal fluxes of these AA showed positive correlations with dry matter intake, whereas the net portal fluxes of Asp, Glu, and Gln were unaffected by treatment. Net hepatic fluxes of EAA were not affected by treatment but increased as lactation progressed with both treatments. On a net basis, all EAA were removed by the liver prepartum and at 4 DIM, whereas Met, Phe, and Thr were the only EAA being removed at 29 DIM. Except for Ala, AA removed by the liver might be used primarily for noncatabolic processes, as exemplified by the 16% of hepatic Gly uptake accounted for as urinary hippurate. The measured hepatic uptake of glucogenic precursors (glucogenic AA, volatile fatty acids, lactate, and glycerol) accounted for 50 to 90% of the hepatic release of glucose. The hepatic urea output accounted for more than 100% of the hepatic ureagenic precursor uptake, indicating that the glucogenic precursors unaccounted for are nonnitrogen-containing compounds. In conclusion, an increased exogenous glucose supply to the small intestine did not seem to affect the amount of EAA and non-EAA available for peripheral tissues in early lactation, and the study did not indicate an AA-sparing effect of small intestinal glucose absorption. In periparturient dairy cows, hepatic catabolism of AA was not driven by the increased whole-body demand for glucose, and Ala was the only AA that contributed substantially to hepatic gluconeogenesis. In very early lactation, the supply of EAA might be of greater concern than the supply of glucogenic substrates.     

Effect of abomasal glucose infusion on plasma concentrations of gut peptides in periparturient dairy cows

Six Holstein cows fitted with ruminal cannulas and permanent indwelling catheters in the portal vein, hepatic vein, mesenteric vein, and an artery were used to study the effects of abomasal glucose infusion on splanchnic plasma concentrations of gut peptides. The experimental design was a randomized block design with repeated measurements. Cows were assigned to one of 2 treatments: control or infusion of 1,500 g of glucose/d into the abomasum from the day of parturition to 29 d in milk. Cows were sampled 12 ± 6 d prepartum and at 4, 15, and 29 d in milk. Concentrations of glucose-dependent insulinotropic polypeptide, glucagon-like peptide 1(7–36) amide, and oxyntomodulin were measured in pooled samples within cow and sampling day, whereas active ghrelin was measured in samples obtained 30 min before and after feeding at 0800 h. Postpartum, dry matter intake increased at a lower rate with infusion compared with the control. Arterial, portal venous, and hepatic venous plasma concentrations of the measured gut peptides were unaffected by abomasal glucose infusion. The arterial, portal venous, and hepatic venous plasma concentrations of glucose-dependent insulinotropic polypeptide and glucagon-like peptide 1(7–36) amide increased linearly from 12 d prepartum to 29 d postpartum. Plasma concentrations of oxyntomodulin were unaffected by day relative to parturition. Arterial and portal venous plasma concentrations of ghrelin were lower postfeeding compared with prefeeding concentrations. Arterial plasma concentrations of ghrelin were greatest prepartum and lowest at 4 d postpartum, giving a quadratic pattern of change over the transition period. Positive portal venous-arterial and hepatic venous–arterial concentration differences were observed for glucagon-like peptide 1(7–36) amide. A negative portal venous–arterial concentration difference was observed for ghrelin pre-feeding. The remaining portal venous–arterial and hepatic venous–arterial concentration differences of gut peptides did not differ from zero. In conclusion, increased postruminal glucose supply to postpartum transition dairy cows reduced feed intake relative to control cows, but did not affect arterial, portal venous, or hepatic venous plasma concentrations of gut peptide hormones. Instead, gut peptide plasma concentrations increased as lactation progressed. Thus, the lower feed intake of postpartum dairy cows receiving abomasal glucose infusion was not attributable to changes in gut peptide concentrations.     

Comparison of net portal absorption with predicted flow of digestible amino acids: scope for improving current models?

This study was undertaken to determine the relationship between measured net portal absorptions (NPA) and flows of digestible essential amino acids (EAA) predicted with the National Research Council model (NRC, 2001) or the Cornell Net Carbohydrate and Protein System model (CNCPS, version 5.0.34). Net portal absorption data were obtained from 33 measurements of portal-arterial plasma EAA concentration differences among 8 treatments in lactating dairy cows, with plasma flow estimated from downstream dilution of para amino-hippurate. The predicted digestible flows from NRC (2001) related better than CNCPS to NPA observed in our studies, as shown by the lower standard errors on the slopes for all EAA and lower root mean prediction errors for all EAA except Met and Phe. However, the partitioning of the prediction error indicated a systematic underprediction (mean bias) for the NRC model (2001), with the exception of Ile. It is important to note that a relationship of unity was not expected, as discussed in the paper, because of losses of EAA through portal-drained viscera metabolism. A revised set of predictive equations for digestible EAA was obtained using a subset of data from NRC (2001) limited to trials conducted with dairy cows. This increased the predicted flows of digestible EAA by only 2%. Flows of digestible EAA were also estimated using a factorial approach, assuming an AA composition for each fraction of the duodenal flow estimated by NRC (undegradable, microbial, and endogenous proteins). This resulted in a slight improvement in the slope of the regression between predicted flows and measured NPA, but still yielded predicted digestive flows that were too low to support observed NPA. Finally, on the basis of literature values, increment of the digestibility of the undegradable fraction of forages and of microbial protein is suggested to improve the relationship between predicted digestible flows and NPA. Overall, this study indirectly confirms, across EAA, smaller losses through gut metabolism for His, Met, and Lys, intermediate losses for the branched-chain AA with the higher losses for Thr.     

Comparison of feed evaluation models on predictions of milk protein yield on Québec commercial dairy farms

Feed evaluation models (FEM) are a core part in dairy cow feeding. As these models are developed using different biological and mathematical approaches mainly tested in a research context, their abilities to predict production in commercial farms need to be validated, even more so when they are used outside the context of their development. Four FEM—National Research Council, 2001 (NRC_2001); Cornell Net Carbohydrate and Protein System, 2015 (CNCPS); NorFor, 2011; and INRA, 2018 (INRA_2018)—were evaluated on their abilities to predict daily milk protein yield (MPY) of 541 cows from 23 dairy herds in the province of Québec, Canada. The effects of cow and diet characteristics were tested on the residuals of MPY. Sensitivity and uncertainty analyses were then performed to evaluate the influence of the uncertainty of the main characteristics of cows and feed ingredients measured on the farm and used in the 4 FEM on the predictions of metabolizable protein (MP) supply and MPY. The 4 models had acceptable predictions of MPY, with concordance correlation coefficients (CCC) ranging from 0.75 to 0.82 and total bias ranging from 12.8% to 19.3% of the observed mean. The Scandinavian model NorFor had the best predictions with a CCC of 0.82, whereas the 3 other models had similar CCC at 0.75 to 0.76. The INRA_2018 and NRC_2001 models presented strong central tendency biases. Removing herd effect put the 4 FEM at the same level of performance, with 11.9 to 12.4% error. Analyzing model behavior within a herd seems to partly negate the effect of using predicted dry matter intake (DMI) in the comparison of models. Diet energy density, days in milk, and MPY estimated breeding value were related to the residual in the 4 models, and Lys and Met (as percent of MP) only in NRC_2001 and NorFor. This suggests that inclusion of these factors in these models would improve MPY predictions. From the sensitivity analysis, for the 4 FEM, DMI and factors affecting its prediction had the greatest influence on the predictions of MP supply and MPY. Of the feed ingredients, forage composition had the greatest effect on these predictions, including a strong effect of legume proportion with NorFor. Diet acid detergent fiber concentration had a very strong effect on MP supply and MPY predictions only in INRA_2018, because of its effect on organic matter digestibility estimation. The range of predictions of MP supply and MPY when combining all these potential uncertainties varied depending on the models. The INRA_2018 model presented the lowest standard deviation (SD) and NorFor the highest SD for the predictions of both MP supply and MPY. Overall, despite the fact that FEM were developed in a research context, their use in a commercial context yields acceptable predictions, with NorFor yielding the best predictions overall, although within-herd responses varied similarly for the 4 tested models.     

Effects of supplementation with 2-hydroxy-4-(methylthio)-butanoic acid isopropyl ester on splanchnic amino acid metabolism and essential amino acid mobilization in postpartum transition Holstein cows

The present study aimed to investigate the effects of 2-hydroxy-4-(methylthio)-butanoic acid isopropyl ester (HMBi) supplementation on splanchnic AA metabolism, essential AA (EAA) mobilization, and plasma AA status in postpartum transition dairy cows. The EAA mobilization was calculated by difference: EAA excretion in milk protein − net portal absorption of EAA or net splanchnic release of EAA. Eight Holstein cows fitted with permanent indwelling catheters in the hepatic portal vein, hepatic vein, mesenteric vein, and an artery in the dry period preceding second parturition were used in the study. Cows were randomly allocated to 1 of 4 treatments in a 2 × 2 factorial arrangement with factor 1: control (calcium carbonate) versus HMBi [1.5 g of HMBi/kg of dry matter (DM)] and factor 2: high dietary ethanol (19 g/kg of DM) versus high dietary propanol (16 g/kg of DM). Only factor 1 data are presented. Treatments were administered in 4 total mixed rations and initiated on the day of parturition. Cows were sampled 14 d before expected parturition and 4, 15, and 29 d after parturition. Supplementation with HMBi tended to increase milk fat content but not fat yield, tended to impose a slower rate of decrease in milk casein content with increasing days in milk (DIM), prevented the decrease in plasma Met associated with parturition for control, reduced plasma concentration of Ser, tended to reduce plasma concentrations of Gly and His, and tended to increase hepatic uptake of Met postpartum. Cows excreted 248 ± 18 g more EAA in the milk at 4 DIM than was released from splanchnic tissues. The EAA deficiency decreased as lactation progressed and was not affected by HMBi supplementation. It was estimated that 4,700 ± 600 g of EAA from extra-splanchnic tissues were secreted in milk protein during the first 29 DIM. Extra-splanchnic EAA mobilization can be crucial to sustain milk protein yield in the postpartum transition period and HMBi is a fast-working Met source that can improve Met status of postpartum transition cows.     

Evaluation of Molly model predictions of ruminal fermentation,nutrient digestion,and performance by dairy cows consuming ryegrass-based diets

Several studies have been conducted to improve grazing management and supplementation in pasture-based systems. However, it is necessary to develop tools that integrate the available information linking the representation of biological processes with animal performance for use in decision making. The objective of this study was to evaluate the precision and accuracy of the Molly cow model predictions of ruminal fermentation, nutrient digestion, and animal performance by cows consuming pasture-based diets to identify model strengths and weaknesses, and to derive new digestive parameters when relevant. Model modifications for adipose tissue, protein synthesis in lean body mass and viscera representation were included. Data used for model evaluations were collected from 25 publications containing 115 treatment means sourced from studies conducted with lactating dairy cattle. The inclusion criteria were that diets contained ≥45% perennial ryegrass (Lolium perenne L.), and that dry matter intake, dietary ingredient composition, and nutrient digestion observations were reported. Animal performance and N excretion variables were also included if they were reported. Model performance was assessed before and after model reparameterization of selected digestive parameters, global sensitivity analysis was conducted after reparameterization, and a 5-fold cross evaluation was performed. Although rumen fermentation predictions were not significantly improved, rumen volatile fatty acids absorption rates were recalculated, which improved the concordance correlation coefficient (CCC) for rumen propionate and ammonia concentration predictions but decreased CCC for acetate predictions. Similar degradation rates of crude protein were observed for grass and total mixed ration diets, but rumen-undegradable protein predictions seemed to be affected by the solubility of the protein source as was the intestinal digestibility coefficient. Ruminal fiber degradation was greater after reparameterization, driven primarily by hemicellulose degradation. Predictions of ruminal and fecal outflow of neutral detergent fiber and acid detergent fiber, as well as total fecal output predictions, improved significantly after reparameterization. Blood urea N and urinary N excretion predictions resulted in similar accuracy using both sets of model parameters, whereas fecal N excretion predictions were significantly improved after reparameterization. Body weight and body condition score predictions were greatly improved after model modifications and reparameterization. Before reparameterization, yield predictions for daily milk, milk fat, milk protein, and milk lactose were greatly overestimated (mean bias of 61.0, 58.7, 73.7, and 64.6% of mean squared error, respectively). Although this problem was partially addressed by model modifications and reparameterization (mean bias of 3.2, 1.1, 1.7, and 0.4% of mean squared error, respectively), CCC values were still small. The ability of the model to predict grass digestion and animal performance in dairy cows consuming pasture-based diets was improved, demonstrating the applicability of this model to these productive systems. However, the failure to predict grass digestion based on standard model inputs without reparameterization indicates there are still fundamental challenges in characterizing feeds for this model.     

Comparison of Molly and Karoline models to predict methane production in growing and dairy cattle

Numerous empirical and mechanistic models predicting methane (CH₄) production are available. The aim of this work was to evaluate the Molly cow model and the Nordic cow model Karoline in predicting CH₄ production in cattle using a data set consisting of 267 treatment means from 55 respiration chamber studies. The dietary and animal characteristics used for the model evaluation represent the range of diets fed to dairy and growing cattle. Feedlot diets and diets containing additives mitigating CH₄ production were not included in the data set. The relationships between observed and predicted CH₄ (pCH₄) were assessed by regression analysis using fixed and mixed model analysis. Residual analysis was conducted to evaluate which dietary factors were related to prediction errors. The fixed model analysis showed that the Molly predictions were related to the observed data (± standard error) as CH₄ (g/d) = 0.94 (±0.022) × pCH₄ (g/d) + 31 (±6.9) [root mean squared prediction error (RMSPE) = 45.0 g/d (14.9% of observed mean), concordance correlation coefficient (CCC) = 0.925]. The corresponding equation for the Karoline model was CH₄ (g/d) = CH₄ (g/d) = 0.98 (±0.019) × pCH₄ (g/d) + 7.0 (±6.0) [RMSPE = 35.0 g/d (11.6%), CCC = 0.953]. Proportions of mean squared prediction error attributable to mean and linear bias and random error were 10.6, 2.2, and 87.2% for the Molly model, and 1.3, 0.3, and 98.6% for the Karoline model, respectively. Mean and linear bias were significant for the Molly model but not for the Karoline model. With the mixed model regression analysis RMSPE adjusted for random study effects were 10.9 and 7.9% for the Molly model and the Karoline model, respectively. The residuals of CH₄ predictions were more strongly related to factors associated with CH₄ production (feeding level, digestibility, fat concentrations) with the Molly model compared with the Karoline model. Especially large mean (underprediction) and linear bias (overprediction of low digestibility diets relative to high digestibility diets) contributed to the prediction error of CH₄ yield with the Molly model. It was concluded that both models could be used for prediction of CH₄ production in cattle, but Karoline was more accurate and precise based on smaller RMSPE, mean bias, and slope bias, and greater CCC. The importance of accurate input data of key variables affecting diet digestibility is emphasized.     

Mammary gland utilization of amino acids and energy metabolites differs when dairy cow rations are isoenergetically supplemented with protein and fat

Mammary gland utilization of AA and other metabolites in response to supplemental energy from protein (PT) and supplemental energy from fat (FT) was tested in a 2 × 2 factorial arrangement using a randomized complete block design. Fifty-six Holstein-Friesian dairy cows were adapted during a 28-d control period to a basal total mixed ration consisting of 34% grass silage, 33% corn silage, 5% grass hay, and 28% concentrate on a dry matter (DM) basis. Experimental rations were fed for 28 d immediately following the control period and consisted of (1) low protein, low fat (LP/LF), (2) high protein, low fat (HP/LF), (3) low protein, high fat (LP/HF), and (4) high protein, high fat (HP/HF). To obtain the high-protein (HP) and high-fat (HF) diets, intake of the basal ration was restricted and supplemented isoenergetically [net energy (MJ/d) basis] with 2.0 kg/d rumen-protected protein (soybean + rapeseed, 50:50 mixture on a DM basis) and 0.68 kg/d hydrogenated palm fatty acids on a DM basis. Arterial and venous blood samples were collected on d 28 of both periods. Isoenergetic supplements (MJ/d) of protein and fat independently and additively increased milk yield, PT increased protein yield, and FT increased fat yield. A PT × FT interaction affected arterial concentration of all essential AA (EAA) groups, where they increased in response to PT by a greater magnitude at the LF level (on average 35%) compared with the HF level (on average 14%). Mammary gland plasma flow was unaffected by PT or FT. Supplementation with PT tended to decrease mammary clearance of total EAA and decreased group 1 AA clearance by 19%. In response to PT, mammary uptake of total EAA and group 2 AA increased 12 and 14%, respectively, with significantly higher uptake of Arg, Ile, and Leu. Energy from fat had no effect on mammary clearance or uptake of any AA group. The mammary gland uptake:milk protein output ratio was not affected by FT, whereas PT increased this ratio for EAA and group 2 AA. Arterial plasma insulin concentration decreased in response to FT, in particular on the HP/HF diet, as indicated by a PT × FT interaction. Arterial concentrations of nonesterified fatty acids, triacylglycerol, and long-chain fatty acids increased in response to FT, and concentrations of β-hydroxybutyrate and acetate decreased in response to FT only at the HP level. Mammary clearance and uptake of triacylglycerol and long-chain fatty acids increased in response to FT. Energy from PT and FT increased lactose yield despite no change in arterial glucose concentration or mammary glucose uptake. Mammary-sequestered glucose with PT or FT was used in the same amount for lactose synthesis, and a positive net mammary glucose balance was found across all treatments. Results presented here illustrate metabolic flexibility of the mammary gland in its use of aminogenic versus lipogenic substrates for milk synthesis.