Responses to amino acid imbalances and deficiencies in lactating dairy cows

Lactating cows were exposed to large amino acid imbalances and deficiencies by i.v. infusion to characterize responses in milk production and plasma concentrations of metabolites and hormones. Six cows in early lactation were fed a basal diet of 9% CP and infused continuously for 6 d with saline (negative control), 1.1 kg/d of a complete amino acid mix (positive control), or the equivalent mix lacking Met, Lys, His, or all 3 branched-chain amino acids. All cows received all treatments in 6 successive periods in a Latin square design. Infusion of the complete amino acid mix resulted in an increase in the plasma concentrations of several essential amino acids, insulin, and glucagon. Milk protein production was stimulated by 19%, which accounted for 10% of the infused amino acid. Plasma urea, acetate, and β-hydroxybutyrate concentrations were increased. Compared with saline, the amino acid mixtures lacking Met, Lys, or His increased essential amino acids, glucose, insulin, and glucagon concentrations in plasma, and decreased growth hormone. Plasma concentration of the essential amino acid absent from the infusate fell 2-fold but milk protein yield remained within 12% of its basal value. Dry matter intakes were depressed 35% over the first 2 d of infusion of imbalanced mixtures but recovered thereafter. Milk fat yields were increased 258 and 320 g/d by mixtures devoid of Lys and His, respectively. Correction of a Met, Lys, or His deficiency did not affect hormone concentrations in plasma and milk protein yield increased 27% due entirely to increased concentration of the single amino acid in plasma. Although imbalance and deficiency generated similar amino acid profiles in plasma, it was concluded that endocrine responses to total amino acid supply during imbalance can override imperfections in the circulating amino acid profile to maintain milk protein yield at higher levels than expected from deficiency states. Both imbalance and deficiency were characterized by a low protein:fat ratio in milk. Infusion of a mix of amino acids lacking Val, Ile, and Leu, despite a decrease in plasma Leu to 58% of its basal value, increased milk protein and fat yields to the same extent as the complete amino acid mix.     

Histidine dose-response effects on lactational performance and plasma amino acid concentrations in lactating dairy cows: 1. Metabolizable protein-adequate diet

The objective of this experiment was to determine the effect of increasing digestible His (dHis) doses on milk production, milk composition, and plasma AA concentrations in lactating dairy cows fed diets that meet or exceed their energy and metabolizable protein (MP) requirements. In a companion paper (Räisänen et al., 2021) results are presented on the effect of increasing dHis dose with an MP-deficient basal diet. In this experiment, 16 Holstein cows (72 ± 15 d in milk) were used in a replicated 4 × 4 Latin square design experiment with four 28-d periods. Treatments were as follows: (1) control, total mixed ration (TMR) with 1.8% dHis of MP (TMR1; dHis1.8); (2) a different TMR with 2.2% dHis (TMR2; dHis2.2); (3) TMR2 supplemented with rumen-protected His (RP-His) to supply 2.6% dHis (dHis2.6); and (4) TMR2 supplemented with RP-His to supply 3.0% dHis of MP (dHis3.0). Estimated dHis intakes calculated at the end of the experiment were 46, 58, 69, and 79 g/d for dHis1.8, dHis2.2, dHis2.6, and dHis3.0, respectively. Contrasts were used to compare TMR1 with TMR2 and to test the linear and quadratic effects of RP-His inclusion rate on TMR2. We detected no effects of TMR or dHis dose on dry matter intake or milk yield, whereas energy-corrected milk (ECM) yield was quadratically increased, being greatest for cows on treatment dHis2.6. Milk true protein and lactose concentrations and milk true protein yield were not affected by TMR or dHis dose. Milk fat concentration and yield increased quadratically, and lactose yield tended to increase quadratically with increasing dHis dose. Calculated apparent efficiency of His utilization decreased quadratically with increasing dHis supply. Further, plasma concentration of His was greater for cows on TMR2 compared with TMR1. When an MP-adequate diet was fed to dairy cows, milk true protein concentration and yield were not affected by dHis supply, but milk fat and ECM yields of dairy cows were optimized at dHis supply of 69 g/d or 2.65% of MP.     

Effects of rumen-protected methionine and other essential amino acid supplementation on milk and milk component yields in lactating Holstein cows

Objectives of this study were to investigate the effects of supplementing rumen-protected methionine (RP-Met), threonine (RP-Thr), isoleucine (RP-Ile), and leucine (RP-Leu) individually or jointly to a low-protein diet, on the performance of lactating dairy cows, as well as to determine the effects of these amino acids (AA) on the mammalian target of rapamycin (mTOR) in vivo. Ten lactating Holstein cows were randomly allocated to a repeated 5 × 5 Latin square experiment with five 19-d periods. Treatments were high-protein diet (16% crude protein, positive control; HP), low-protein diet (12% crude protein, negative control; LP), LP plus RP-Met (LPM), LP plus RP-Met and RP-Thr (LPMT), and LP plus RP-Met, RP-Thr, RP-Ile, and RP-Leu (LPMTIL). The dry matter intakes (DMI) of the LP, LPM, and LPMT diets were lower than that of the HP diet, whereas the DMI of the LPMTIL diet was intermediate between the HP diet and the other LP diets. Supplementing RP-Met to the LP diet increased the yields of milk and milk protein, increased the content of milk urea N, and tended to increase milk N efficiency. Co-supplementation of RP-Thr with RP-Met resulted in no further milk production increase. Co-supplementation of all 4 rumen-protected amino acids (RP-AA) increased milk and lactose yields to the level of the HP diet and tended to increase milk protein yield compared with the LPMT diet. We found no significant differences in the contents and yields of milk components between the LPMTIL and HP diets except for a lower milk urea N content in the LPMTIL diet. Venous concentrations of the measured AA were similar across the LP and LP diets supplemented with RP-AA. Relative to levels of the HP diet, LP diets had higher venous concentrations of Met and Gly and tended to have higher Phe concentration and lower concentrations of Val and BCAA. The LPMTIL diet had higher venous concentrations of Arg, Lys, Met, Phe, and Glu, and a lower Val concentration. Phosphorylation status of the measured mTOR components in LPM and LPMT treatments were similar to those in the LP treatment but phosphorylation status of mTOR and eIF4E-binding protein 1 (4eBP1) in LPMTIL treatment were higher. The phosphorylation rates of eukaryotic elongation factor 2 (eEF2) in the 4 LP and LP plus RP-AA diets were higher than that of the HP diet. Overall, results of the present study supported the concept that under the relatively short time of this experiment, supplementing RP-AA, which are believed to stimulate the mTOR signal pathway, can lead to increased milk protein yield. This increase appears to be due to increased DMI, greater mTOR signaling, and greater eEF2 activity.     

Effects of reduced dietary protein and supplemental rumen-protected essential amino acids on the nitrogen efficiency of dairy cows

When fed to meet the metabolizable protein requirements of the National Research Council, dairy cows consume an excess of N, resulting in approximately 75% of dietary N being lost to the environment as urine and feces. Reductions in environmental N release could be attained through an improvement in N efficiency. The objective of this study was to determine if the predicted reduction in milk yield associated with feeding a low-protein diet to lactating dairy cows could be avoided by dietary supplementation with 1 or more ruminally protected (RP) AA. Fourteen multiparous and 10 primiparous Holstein cows, and 24 multiparous Holstein × Jersey crossbred cows were used in a Youden square design consisting of 8 treatments and 3 periods. The 8 dietary treatments were (1) a standard diet containing 17% crude protein [CP; positive control (PC)], (2) a 15% CP diet [negative control (NC)], (3) NC plus RP Met (+M), (4) NC plus RP Lys (+K), (5) NC plus RP Leu (+L), (6) NC plus RP Met and Lys (+MK), (7) NC plus RP Met and Leu (+ML), and (8) NC plus RP Met, Lys, and Leu (+MKL). Dry matter intake was not affected by treatment. Crude protein intake was lower for NC and RP AA treatments compared with the PC treatment. No detrimental effect was detected of the low-CP diet alone or in combination with AA supplementation on milk and fat yield. However, milk protein yield decreased for NC and +MKL diets, and lactose yield decreased for the +MKL compared with the PC diet. Milk urea N concentrations were lower for all diets, suggesting that greater N efficiency was achieved by feeding the low-protein diet. Minimal effects of treatments on arterial plasma essential AA concentrations were detected, with only Ile and Val being significantly lower in the NC than in the PC diet. Phosphorylation ratios of signaling proteins known to regulate mRNA translation were not affected by treatments. This study highlights the limitations of requirement models aggregated at the protein level and the use of fixed postabsorptive efficiency to calculate milk protein requirements. Milk protein synthesis regulation by signaling pathways in vivo is still poorly understood.     

Lactational performance of dairy cows in response to supplementing N-acetyl-l-methionine as source of rumen-protected methionine

The objective of this experiment was to evaluate the effects of supplementing a rumen-protected source of Met, N-acetyl-l-methionine (NALM), on lactational performance and nitrogen metabolism in early- to mid-lactation dairy cows. Sixty multiparous Holstein dairy cows in early lactation (27 ± 4.3 d in milk, SD) were assigned to 4 treatments in a randomized complete block design. Cows were blocked by actual milk yield. Treatments were as follows: (1) no NALM (control); (2) 15 g/d of NALM (NALM15); (3) 30 g/d of NALM (NALM30); and (4) 45 g/d of NALM (NALM45). Diets were formulated using a Cornell Net Carbohydrate and Protein System (CNCPS) v.6.5 model software to meet or exceed nutritional requirements of lactating dairy cows producing 42 kg/d of milk and to undersupply metabolizable Met (control) or supply incremental amounts of NALM. The digestible Met (dMet) supply for control, NALM15, NALM30, and NALM45 were 54.7, 59.8, 64.7, and 72.2 g/d, respectively. The supply of dMet was 88, 94, 104, and 115% of dMet requirement for control, NALM15, NALM30, and NALM45, respectively. Milk yield data were collected, dry matter intake (DMI) was measured daily, and milk samples were collected twice per week for 22 wk. Blood, ruminal fluid, urine, and fecal samples were collected during the covariate period and during wk 4, 8, and 16. Data were analyzed using the GLIMMIX procedure of SAS (SAS Institute) using covariates in the model for all variables except body weight. Linear, quadratic, and cubic contrasts were also tested. Treatments did not affect DMI, milk yield, and milk component concentration and yield; however, feed efficiency expressed as milk yield per DMI and 3.5% fat-corrected milk per DMI were quadratically affected, with greater response observed for NALM15 and NALM30 compared with control. Acetate proportion linearly increased, whereas propionate proportion linearly decreased with NALM supplementation. Blood urea nitrogen linearly decreased with NALM supplementation. Total plasma essential AA concentrations were quadratically affected, as greater values were observed for control and NALM45 than other treatments. Plasma Met concentration was quadratically affected as lower levels were observed with NALM15, whereas Met concentrations increased with NALM45 compared with control. Nitrogen utilization efficiency and apparent total-tract nutrient digestibility were not affected by treatment. Supplementation of NALM at 15 or 30 g/head per day resulted in the greatest improvements in feed efficiency without affecting N metabolism of early- to mid-lactation dairy cows.     

Plasma essential amino acid concentrations in response to casein infusion or ration change in dairy cows: A multilevel,mixed-effects meta-analysis

The objectives of the current meta-analysis were to review the relationships between plasma individual essential AA concentrations and their respective AA digestible flows (AADI) in 2 independent sets of studies. The first set of studies included 36 casein infusion studies (CN; 83 treatment means) and was regarded as the best comparison standard available, because differences in AADI achieved by casein infusion (up to 40% of total metabolizable protein supply) did not rely on any model assumptions and were directly estimated from casein infusions. The second set of studies included 42 feeding trials (FT; 94 treatment means) in which AADI were predicted using the 2001 National Research Council model. The 2 sets of studies were not balanced for dry matter intake and the supplies of metabolizable protein and net energy for lactation; therefore, a subset of 17 CN studies (35 treatment means) and 19 FT trials (49 treatment means) balanced for these variables was assembled to allow the comparison of linear terms from CN and FT studies. In the subset of data set, the linear terms of individual AA did not differ between CN and FT studies except for Met and Thr, with a slope lower by 23 and 62%, respectively, in CN versus FT studies. The agreement in linear slopes between CN and FT studies indicates, indirectly, that AADI were predicted accurately by the National Research Council model. In the large data set, the relationships between plasma concentrations of Ile, Leu, Val, and their sum (branched-chain AA; BCAA) and their respective AADI shared common characteristics that distinguished them from the other AA. For the plasma concentration of BCAA, the linear terms were significant in CN and FT studies, but the quadratic terms were significant only in FT studies. This decline in the response of plasma concentration of BCAA to increased BCAA digestible flow in FT studies was associated with diets rich in energy, diets with a high concentrate level, or diets based on corn silage. These dietary conditions can stimulate insulin secretion and decrease plasma concentration of BCAA. For the non-BCAA, a quadratic term was significant for plasma His, Lys, Met, and Thr in each set of studies, indicating an increased removal of these AA by the liver as AADI increased.     

Effects of corn silage processing and amino acid supplementation on the performance of lactating dairy cows

This experiment was conducted to determine the effect of crop processing and amino acid supplementation on dairy cow performance. Corn silage processed (PCS) or unprocessed (UCS) was used as the main forage (45% of dry matter, DM) in a total mixed ration (TMR). Each TMR was either supplemented (AA) or not (AAO) with ruminally protected amino acids (lysine, 3 g/d and methionine, 14 g/d). Thirty-two (551 kg) Holstein cows were randomly assigned to four treatments: PCS-AA, PCS-AA0, UCS-AA, and UCS-AA0 in a 2 x 2 factorial structure. Between wk 7 and 17 of lactation, cows were fed ad libitum TMR comprising 45% of corn silage plus 1 kg of grass hay once a day. The UCS presented better fermentation characteristics than PCS. Dry matter intake (DMI) of the TMR was not affected by treatment and averaged 22.7 kg/d. Energy-corrected milk (ECM) production was 9% higher with UCS than with PCS (33.1 vs. 30.1 kg/d). Milk efficiency was therefore 6% higher with UCS than with PCS (1.43 vs. 1.35 kg ECM/kg of DMI). The concentration of major milk constituents (fat, protein, lactose, urea) was not affected by treatments. Apparent digestibility of DM, organic matter, N, starch, acid detergent fiber, and neutral detergent fiber were similar among treatments. The effective ruminal degradability of DM, starch, and protein, however, was greater with PCS than with UCS. Amino acid supplementation had no effect on milk production nor on milk constituents, whether it was used with processed corn silage or with unprocessed corn silage. These data indicate that feeding UCS resulted in a greater milk production compared with PCS. The numerically higher DMI, a potentially greater intestinal digestion of starch or the better conservation of UCS could have contributed to the greater milk production.     

Urea-N recycling in lactating dairy cows fed diets with 2 different levels of dietary crude protein and starch with or without monensin

Rumensin (monensin; Elanco Animal Health, Greenfield, IN) has been shown to reduce ammonia production and microbial populations in vitro; thus, it would be assumed to reduce ruminal ammonia production and subsequent urea production and consequently affect urea recycling. The objective of this experiment was to determine the effects of 2 levels of dietary crude protein (CP) and 2 levels of starch, with and without Rumensin on urea-N recycling in lactating dairy cattle. Twelve lactating Holstein dairy cows (107 ± 21 d in milk, 647 kg ± 37 kg of body weight) were fed diets characterized as having high (16.7%) or low (15.3%) CP with or without Rumensin, while dietary starch levels (23 vs. 29%) were varied between 2 feeding periods with at least 7 d of adaptation between measurements. Cows assigned to high or low protein and to Rumensin or no Rumensin remained on those treatments to avoid carryover effects. The diets consisted of approximately 40% corn silage, 20% alfalfa hay, and 40% concentrate mix specific to the treatment diets, with 0.5 kg of wheat straw added to the high starch diets to enhance effective fiber intake. The diets were formulated using Cornell Net Carbohydrate and Protein System (version 6.1), and the low-protein diets were formulated to be deficient for rumen ammonia to create conditions that should enhance the demand for urea recycling. The high-protein diets were formulated to be positive for both rumen ammonia and metabolizable protein. Rumen fluid, urine, feces, and milk samples were collected before and after a 72-h continuous jugular infusion of ¹⁵N¹⁵N-urea. Total urine and feces were collected during the urea infusions for N balance measurements. Milk yield and dry matter intake were improved in cows fed the higher level of dietary CP and by Rumensin. Ruminal ammonia and milk and plasma urea nitrogen concentrations corresponded to dietary CP concentration. As has been shown in vitro, Rumensin reduced rumen ammonia concentration by approximately 23% but did not affect urea entry rate or gastrointestinal entry rate. Urea entry rate averaged approximately 57% of total N intake for cattle with and without Rumensin, and gastrointestinal rate was similar at 43 and 42% of N intake for cattle fed and not fed Rumensin, respectively. The cattle fed the high-protein diet had a 25% increase in urea entry rate and no effect of starch level was observed for any recycling parameters. Contrary to our hypothesis, Rumensin did not alter urea production and recycling.     

Effects of dietary crude protein concentration on late-lactation dairy cow performance and indicators of nitrogen utilization

The objectives of this study were to measure performance responses and to evaluate indictors of N utilization in late-lactation cows fed diets with incremental reductions in crude protein (CP) concentration. Holstein cows (n = 128; 224 ± 54 d in milk) were stratified by parity and days pregnant (86 ± 25 d) and randomly assigned to 1 of 16 pens in a randomized complete block design. For 3 wk, all cows received a covariate diet containing 16.9% CP [dry matter (DM) basis]. For the subsequent 12 wk, pens were randomly assigned to 1 of 4 treatments that contained 16.2, 14.4, 13.1, or 11.8% CP (DM basis). Diets were offered once daily and contained 32.5% corn silage, 32.5% alfalfa silage, 13.5% high-moisture corn, and 21.5% concentrate mix. A reduction in dietary CP was achieved by replacing soybean meal with soy hulls in the concentrate mix (DM basis). Dry matter intake, milk urea N (MUN; mg/dL), and the yield of milk urea N (g/d) decreased linearly with dietary CP. Compared with a 16.2% CP diet, a 14.4% CP diet did not alter milk yield throughout the study, but the 13.1 and 11.8% CP diets reduced milk yield after 4 and 1 wk, respectively. Furthermore, milk protein percentage was reduced for all dietary CP less than 16.2%, but this negative effect was temporary and disappeared after 7 wk for the 14.4% CP diet. In contrast, MUN adjusted to a new steady state within 1 wk for all dietary treatments. Modeling quadratic responses with a plateau led to predictions of no reduction in fat- and protein-corrected milk (32.6 kg/d) and yields of fat (1.31 kg/d), lactose (1.49 kg/d), and true protein (1.12 kg/d) until dietary CP decreased below 15.5, 15.3, 15.9, and 16.2%, respectively. In this study, MUN and the yield of MUN were highly correlated with N intake, milk protein yield, and fat- and protein-corrected milk. Surprisingly, N use efficiency (milk protein N/intake N) was not correlated with any variables related to N utilization and reached an apparent upper limit of approximately 30%. Although this observation may be associated with feeding diets deficient in metabolizable protein, late-lactation cows in this study adjusted to low dietary CP concentration better than anticipated as milk production was 2.6, 3.6, 6.4, and 8.0 kg/d higher than National Research Council (2001)-predicted metabolizable protein-allowable milk for dietary CP of 16.2, 14.4, 13.1, and 11.8%, respectively.     

Energy and nitrogen balance of dairy cattle as affected by provision of different essential amino acid profiles at the same metabolizable protein supply

Amino acid composition of metabolizable protein (MP) is important in dairy cattle diets, but effects of AA imbalances on energy and N utilization are unclear. This study determined the effect of different AA profiles within a constant supplemental MP level on whole-body energy and N partitioning in dairy cattle. Five rumen-fistulated Holstein-Friesian dairy cows (2.8 ± 0.4 lactations; 81 ± 11 d in milk; mean ± standard deviation) were randomly assigned to a 5 × 5 Latin square design in which each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of rest. A total mixed ration consisting of 58% corn silage, 16% alfalfa hay, and 26% concentrate (dry matter basis) was formulated to meet 100 and 83% of net energy and MP requirements, respectively, and was fed at 90% of ad libitum intake by individual cow. Abomasal infusion treatments were saline (SAL) or 562 g/d of essential AA delivered in 4 profiles where individual AA content corresponded to their relative content in casein. The profiles were (1) a complete essential amino acid mixture (EAAC), (2) Ile, Leu, and Val (ILV), (3) His, Ile, Leu, Met, Phe, Trp, Val (GR1+ILV), and (4) Arg, His, Lys, Met, Phe, Thr, Trp (GR1+ALT). The experiment was conducted in climate respiration chambers to determine energy and N balance in conjunction with milk production and composition, digestibility, and plasma constituents. Compared with SAL, infusion of EAAC increased milk, protein, and lactose yield, increased energy retained as body protein, and did not affect milk N efficiency. Total N intake and urine N output was higher with all AA infusions relative to SAL. Compared with EAAC, infusions of GR1+ILV and GR1+ALT produced the same milk yield and the same yield and content of milk fat, protein, and lactose, and had similar energy and N retention. Milk N efficiency was not different between EAAC and GR1+ILV, but was lower with GR1+ALT compared with EAAC, and tended to be lower with GR1+ALT compared with GR1+ILV. Infusion of ILV tended to decrease dry matter intake compared with the other AA infusions. Milk production and composition was not different between ILV and SAL. Compared with EAAC, infusion of ILV decreased or tended to decrease milk, protein, and lactose yields and milk protein content, and increased milk fat and lactose content. Milk N efficiency decreased with ILV compared with SAL, EAAC, and GR1+ILV. Milk urea concentration was not affected by essential amino acid (EAA) infusions. Plasma urea concentration did not differ between EAAC and SAL, tended to increase with ILV and GR1+ILV over SAL, and increased with GR1+ALT compared with EAAC and SAL. In conclusion, removing Arg, Lys, and Thr or removing Ile, Leu, and Val from a complete EAA profile when the total amount of EAA infused remained constant did not impair milk production, but milk N efficiency decreased when Ile, Leu, and Val were absent. Infusion of only Ile, Leu, and Val decreased milk protein yield and content and reduced milk N efficiency compared with a complete EAA profile.     

Changes in production and mammary metabolism of dairy cows in response to essential and nonessential amino acid infusions

This study was undertaken to increase our understanding of the need of the mammary gland for the different types of AA and how the mammary gland alters its metabolism in response to a variable AA supply. Eight lactating Holstein cows (61 ± 4 DIM) were used in a replicated 4 × 4 Latin square balanced for residual effects with 14-d periods. The diet was formulated to supply 100% of the net energy requirement and 72% of the metabolizable protein requirement. The 4 treatments were 1) abomasal infusions of water, 2) essential AA at 359 g/d, 3) nonessential AA at 356 g/d, and 4) essential AA at 359 g/d + nonessential AA at 356 g/d (total of 715 g/d). The infusates had the same AA profile as casein with the exception that Met was increased to maintain a 3:1 ratio of digestible Lys to Met and because of solubility limitations all the Tyr was replaced by Phe and part of the Glu was replaced by Gln. Milk yield and milk protein yield were increased by the essential AA treatments compared with the other treatments. Mammary uptake of β-hydroxybutyrate plus lactate tended to increase with the essential AA treatments, whereas glucose mammary uptake tended to be higher with the nonessential AA treatments. With the essential AA treatments, the mammary uptake:milk protein output ratio for the group 1 AA (His, Met, Phe, Trp, and Tyr) did not differ from 1 but tended to increase; the ratio for the group 2 AA (Ile, Leu, Lys, and Val) did increase, significantly exceeding unity when essential AA were infused. These results indicate that the mammary gland alters differently its uptake of both AA and energy-yielding nutrients in response to the amount and profile of AA presented to it and that even under situations of protein deficiency nonessential AA supplementation does not enhance milk and milk protein synthesis.     

Lowering rumen-degradable and rumen-undegradable protein improved amino acid metabolism and energy utilization in lactating dairy cows exposed to heat stress

The objective of this study was to evaluate the effects of reducing dietary rumen-degradable protein (RDP) and rumen-undegradable protein (RUP) on protein and energy metabolism in heat-stressed dairy cows. Eighteen primiparous and 30 multiparous mid-lactation Holstein cows were used in a completely randomized design arranged in a 2 × 2 factorial (n = 12/treatment). Cows were randomly assigned to 1 of 4 dietary treatments that included 2 levels of RDP (10 and 8%; D) and 2 levels of RUP (8 and 6%; U) of dry matter for 21 d as (1) 10D:8U, (2) 8D:8U, (3) 10D:6U, and (4) 8D:6U. Diets were isoenergetic and contained 50% forage and 50% concentrate (dry matter basis). Cows were housed in a freestall barn. Three weeks before start of treatments, all animals were fed the 10D:8U diet and received supplemental cooling to prevent heat stress. During the treatment period, cows experienced a daily increment in temperature-humidity index from 74 to 82 for 1000 to 2000 h. Blood samples were collected on d ?1 and 21 of the treatment period to determine plasma concentrations of AA, glucose, insulin, fatty acids, and β-hydroxybutyrate. For primiparous cows, reducing from 10 to 8% RDP decreased insulin concentrations. For multiparous cows, we found significant RDP by RUP interactions for insulin, β-hydroxybutyrate, fatty acids, total essential AA, and 3-methylhistidine concentrations. Reducing from 10 to 8% RDP decreased insulin concentrations at 6% RUP, but concentrations did not change when reducing RDP at 8% RUP. Reducing from 10 to 8% RDP decreased β-hydroxybutyrate concentrations at 8% RUP, but concentrations did not change when reducing RDP at 6% RUP. Reducing from 10 to 8% RDP increased nonesterified fatty acid and total essential AA concentrations at 8% RUP, but concentrations did not change when reducing RDP at 6% RUP. Reducing from 8 to 6% RUP decreased 3-methylhistidine concentration at 8% RDP, but not at 10% RDP. Reducing from 8 to 6% RUP increased milk protein yield efficiency in primiparous and multiparous cows. These results indicate that reducing RDP and RUP lowers circulating insulin, which was associated with mobilization and utilization of fatty acids. Reduced RDP and RUP increases the use of AA to maintain milk protein synthesis and limit AA catabolism in cows exposed to warm climates.     

Effect of amino acid or casein supply on whole-body, splanchnic, and mammary glucose kinetics in lactating dairy cows

This study was conducted to establish how AA supplied in a free form or as protein (casein, CN) affect the whole-body rate of appearance (WB Ra) of glucose, splanchnic and mammary glucose kinetics, and milk lactose secretion in lactating dairy cows. Five Holstein cows fitted with a rumen cannula and permanent indwelling catheters in the abomasum, portal, hepatic, and mesenteric veins, and one mesenteric artery, were used in a Youden square with 4 periods of 14 d each. Cows were fed a hay-based diet providing 100 and 70% of their net energy and metabolizable protein (MP) requirements, respectively. Treatments consisted of abomasal infusions of water (70% of MP requirements: control, Con), free AA (95% of MP requirements: AA1; and 120% of MP requirements: AA2), or CN (95% of MP requirements: CN1). The free AA mixture had the same profile as CN. On d 14 of each period, [6,6-²H₂]glucose (25.8 mmol/h) was infused into a jugular vein, and blood samples (n = 8) were taken over 4 h from arterial, portal, hepatic, and mammary sources to measure glucose enrichment and concentration. Splanchnic and mammary plasma flows were determined by downstream dilution of para-aminohippurate and with the Fick principle, respectively. The last 6 milkings of each period were weighed and sampled to measure the yields of milk and components. The AA1 and CN1 treatments were not different for any of the measured parameters. Supplying AA linearly increased glucose WB Ra (AA2 vs. Con: +151 mmol/h) and liver net flux (+149 mmol/h). Utilization of glucose from the plasma compartment by the portal-drained viscera and liver and true portal absorption were not affected by AA supply. From these observations, we suggest that the increased WB Ra was due to increased net hepatic production. The AA from the infusion, in excess of that used to cover the increase in milk protein, were converted to glucose with an apparent efficiency close to 100% of maximum theoretical efficiency. Milk and lactose yields increased linearly with infusions of AA, by 14 and 16% with AA2 treatment, respectively. However, mammary glucose uptake was not significantly altered by AA infusions; this suggests that the mammary gland exerts active control on the uptake and utilization of glucose. For all treatments, the sum of true portal glucose absorption and true hepatic glucose production contributed more than 99% of WB Ra in the lactating cow; this would suggest that renal glucose synthesis makes only a small contribution to WB Ra under these conditions.     

Modeling portal-drained viscera and liver fluxes of essential amino acids in dairy cows

The objective of this work was to predict essential amino acid (EAA) use and release by the portal-drained viscera (PDV) and liver of dairy cows. Previously derived equations were tested using data assembled from the literature, refit to the data, and modifications were undertaken to determine the best model for each EAA. The refitted model has the same structure as the original equations but is parameterized using a database of group means, as the original equations were derived using a single study with individual cow data and found to be biased. The PDV clearance model predicted portal vein concentrations given inputs of absorbed and arterial fluxes of EAA with root mean squared errors (RMSE) ranging from 3.3 to 12.1% of the observed means, and concordance correlation coefficients (CCC) ranging from 0.86 to 0.99 when using previously reported parameters. The reparameterized model generated from the assembled data set resulted in predictions of EAA portal vein concentrations with RMSE ranging from 3.2 to 8.6% and CCC ranging from 0.93 to 1.00. Slope bias ranged from 12.4 to 55.3% of mean squared errors and was correlated with arterial EAA concentrations. Modifying the model to allow rate constants to vary as a function of arterial EAA concentrations reduced slope bias, resulting in RMSE ranging from 1.9 to 6.5% and CCC from 0.97 to 1.00. Alternatively, splitting the model to account for use of EAA from absorption separately from arterial use resulted in poorer predictions and biologically infeasible parameter estimates. The liver clearance model predicted hepatic vein concentrations from arterial and portal vein input fluxes with RMSE across EAA ranging from 1.9 to 6.8% and CCC ranging from 0.97 to 1.00 when using reported parameters. The reparameterized model generated from the assembled data set resulted in predictions of EAA hepatic vein concentrations with RMSE ranging from 1.9 to 6.7% and CCC ranging from 0.97 to 1.00. Significant slope bias was present for Arg, His, Lys, Phe, Thr, and Val. Altering the model to represent the clearance rate constant as a function of arterial concentrations resulted in RMSE ranging from 1.8 to 6.5% and CCC ranging from 0.97 to 1.00. The combination of PDV and liver clearance models provided predictions of total splanchnic use similar to those of an empirical model representing splanchnic use as a fractional proportion of absorption that had RMSE ranging from 3.0 to 8.6% and CCC ranging from 0.95 to 0.99, with significant slope bias for the majority of EAA.     

Regulation of protein synthesis in mammary glands of lactating dairy cows by starch and amino acids

The objective of this study was to evaluate local molecular adaptations proposed to regulate protein synthesis in the mammary glands. It was hypothesized that AA and energy-yielding substrates independently regulate AA metabolism and protein synthesis in mammary glands by a combination of systemic and local mechanisms. Six primiparous mid-lactation Holstein cows with ruminal cannulas were randomly assigned to 4 treatment sequences in a replicated incomplete 4 × 4 Latin square design experiment. Treatments were abomasal infusions of casein and starch in a 2 × 2 factorial arrangement. All animals received the same basal diet (17.6% crude protein and 6.61 MJ of net energy for lactation/kg of DM) throughout the study. Cows were restricted to 70% of ad libitum intake and abomasally infused for 36 h with water, casein (0.86 kg/d), starch (2 kg/d), or a combination (2 kg/d starch + 0.86 kg/d casein) using peristaltic pumps. Milk yields and composition were assessed throughout the study. Arterial and venous plasma samples were collected every 20 min during the last 8 h of infusion to assess mammary uptake. Mammary biopsy samples were collected at the end of each infusion and assessed for the phosphorylation state of selected intracellular signaling molecules that regulate protein synthesis. Animals infused with casein had increased arterial concentrations of AA, increased mammary extraction of AA from plasma, either no change or a trend for reduced mammary AA clearance rates, and no change in milk protein yield. Animals infused with starch had increased milk and milk protein yields, increased mammary plasma flow, reduced arterial concentrations of AA, and increased mammary clearance rates and net uptake of some AA. Infusions of starch increased plasma concentrations of glucose, insulin, and insulin-like growth factor-I. Starch infusions increased phosphorylation of ribosomal protein S6 and endothelial nitric oxide synthase, consistent with changes in milk protein yields and plasma flow, respectively. Phosphorylation of the mammalian target of rapamycin was increased in response to starch only when casein was also infused. Thus, cell signaling molecules involved in the regulation of protein synthesis differentially responded to these nutritional stimuli. The hypothesized independent effects of casein and starch on animal metabolism and cell signaling were not observed, presumably because of the lack of a milk protein response to infused casein.     

Short- and longer-term effects of feeding increased metabolizable protein with or without an altered amino acid profile to dairy cows immediately postpartum

The first few weeks after parturition is marked by low, but increasing feed intake and sharply increasing milk production by dairy cows. Because of low intake, the nutrient density of the diet may need to be higher during this period to support increasing milk yields. We hypothesized that feeding higher levels of metabolizable protein (MP) or a protein supplement with rumen-protected lysine and methionine during the immediate postpartum period would increase yields of milk and milk components. Fifty-six Holstein cows (21 primiparous and 35 multiparous) starting at 3 d in milk were used in a randomized block design. In phase 1 (3 through 23 d in milk), cows were fed 1 of 3 diets that differed in supply of MP and AA profile. At 23 d in milk, all cows were moved to a common freestall pen and fed the control diet used in phase 1 for an additional 63 d (phase 2). Diets were formulated using the National Research Council model and were control [16.5% crude protein (CP), 10.9% rumen-degradable protein (RDP), and 5.6% rumen-undegradable protein (RUP)], high MP (HMP; 18.5% CP, 11.6% RDP, 6.9% RUP), and AA (MPAA; 17.5% CP, 10.5% RDP, 7.0% RUP 29.7). The MPAA diet included a proprietary spray-dried blood meal product (Perdue Agribusiness, Salisbury, MD) and contained a model-estimated 7.2 and 2.6% of digestible lysine and methionine (% of MP). The HMP and control diets contained 6.3 and 6.7% digestible lysine and both had 1.8% digestible methionine. In phase 1, diet did not affect milk yield (33.6, 34.7, and 33.2 kg for control, HMP, and MPAA, respectively), dry matter intake (17.8, 18.0, and 18.5 kg/d for control, HMP, and MPAA), or milk protein yield (1.07 kg/d). Feeding additional protein (HMP or MPAA) increased both the concentration and yield of milk fat, and milk protein concentration was greater (3.30 vs. 3.17%) for MPAA compared with the HMP diet. Energy-corrected milk was greater (38.4 and 38.6 vs. 35.3 kg/d, respectively) for MPAA and HP than for the control. Cows fed MPAA had the greatest plasma concentrations of Met and the lowest concentrations of isoleucine, but lysine was not affected by treatment. Feeding additional MP (HMP or MPAA) reduced the concentrations of 3-methylhistidine in plasma, indicating reduced muscle breakdown. Diet effects on milk composition continued after cows were changed to a common diet in that cows fed MPAA the first 3 wk of lactation had greater concentration of milk protein for the entire experiment than cows fed HMP, and cows fed additional MP (HMP and MPAA) during phase 1 had greater concentrations of milk fat for the entire experiment. Increasing dietary protein and AA supply in early lactation had short-term effects on yield of energy-corrected milk and long-term effects on milk composition.     

Effects of increased doses of lysine in a rumen-protected form on plasma amino acid concentration and lactational performance of dairy cows fed a lysine-deficient diet

The objective of these studies was to determine the effects of feeding a novel rumen-protected Lys (RP-Lys) product on plasma AA, lactational performance, and Lys bioavailability. To evaluate RP-Lys on lactation performance a corn-based diet (42.5% of corn silage and 21.9% of corn and corn by-products, on DM basis) was formulated to be Lys deficient but adequate in Met, energy, and metabolizable protein. Thirty-six lactating Holstein cows were fed either a Lys-deficient control diet (CON) with no added RP-Lys, or diets containing 0.3% of RP-Lys (0.3RP-Lys) or 0.6% of RP-Lys (0.6RP-Lys) for 8 wk. There were no effects on dry matter intake (mean ± SD; 26.1 ± 0.58 kg/d), milk yield (37.9 ± 0.72 kg/d), or milk composition to the RP-Lys supplementation. No effect was observed on plasma AA concentrations except for His. Plasma His was linearly reduced by Lys feeding (42.6, 41.2, 30.0 ± 4.09 μM, for CON, 0.3RP-Lys, and 0.6RP-Lys, respectively). Calculated efficiency of Lys utilization decreased linearly with RP-Lys supplementation. In the companion study, 3 rumen-cannulated lactating dairy cows were used in a 3 × 3 Latin square design to assess the bioavailability of the RP-Lys. Free Lys (HCl-Lys), RP-Lys, and water were administered separately by postruminal bolus dosing. The Lys bioavailability was assessed by the ratio of area under the curve of Lys plasma concentration for RP-Lys compared with HCl-Lys and discounted for the area under the curve for water bolus dose. The estimated bioavailability of the RP-Lys was 24.4% ± 4.61. In summary, increased supplemental doses of Lys had no effect on Lys plasma concentration and lactational performance when fed to dairy cows on a corn-based diet, although altered Lys as % of essential AA was observed. However, the lack of effects should be considered in light of the lower-than-expected bioavailability of the RP-Lys.     

Performance and amino acid utilization of early lactation dairy cows fed regular or reduced-fat dried distillers grains with solubles

The objective of this study was to evaluate lactation response and AA utilization of early lactation cows fed 2 types of dried distillers grains with solubles (DG): regular (DDGS) or reduced-fat (RFDGS). Thirty-six Holstein cows 19.7 ± 2.6 d in milk at the start of the experiment were used in a randomized complete block design for 14 wk including a 2-wk covariate period. Treatments consisted of the following diets: 1) control (CON) diet containing 0% DG; 2) diet containing 22% DDGS; and 3) diet containing 20% RFDGS. Distillers grains replaced soybean meal, expeller soybean meal, and soyhulls from the CON diet. Diets were formulated to be similar in crude protein, ether extract, neutral detergent fiber, and net energy for lactation concentrations. Dry matter intake (24.7 kg/d) and milk yield (39.3 kg/d) were similar for all diets. Milk fat and lactose percentages were unaffected by diets; however, protein percentage was greater for cows fed the DG diets compared with the CON diet. Consequently, milk protein yield was also greater for the DG diets compared with CON. Milk urea nitrogen decreased for cows fed DG diets and averaged 11.8, 10.9, and 10.1 mg/dL, respectively, for CON, DDGS, and RFDGS. Feed efficiency tended to be greater and N efficiency was greater for cows fed DG compared with CON. Body weight (711 kg), body weight change (+0.49 kg/d), and body condition score (3.36) were similar for all diets, but cows fed CON tended to gain more body condition (+0.14) than cows fed DG diets. Amino acid utilization was evaluated at the peak of milk production corresponding to wk 9 of lactation. Arterial Lys concentration was lower with DG diets (70.4, 58.6, and 55.8 μM/L). Cows fed DG had greater arterial Met concentration (21.3 μM) compared with CON (14.9 μM). Arterio-venous difference of Lys was similar across diets, whereas that of Met was greater for the DG diets compared with the CON diet (10.3 vs. 13.0 μM/L). Extraction efficiency of Lys by the mammary gland was greater for DG diets than for CON (76.1 vs. 65.4%). Mammary uptake of Lys (2.56 g/kg of milk) was similar for all diets, and the uptake of Met tended to increase in cows fed DG diets. Plasma glucose, triglyceride, and total cholesterol were unaffected by treatment; however, cows fed DG diets had lower β-hydroxybutyrate and tended to have lower nonesterified fatty acid concentrations than cows fed the CON diet. Despite the apparent deficiency of Lys, milk protein percentage was increased in cows fed DG diets.     

Models to predict milk fat concentration and yield of lactating dairy cows: A meta-analysis

Few models have attempted to predict total milk fat because of its high variation among and within herds. The objective of this meta-analysis was to develop models to predict milk fat concentration and yield of lactating dairy cows. Data from 158 studies consisting of 658 treatments from 2,843 animals were used. Data from several feed databases were used to calculate dietary nutrients when dietary nutrient composition was not reported. Digested intake (DI, g/d) of each fatty acid (FA; C12:0, C14:0, C16:0, C16:1, C18:0, C18:1 cis, C18:1 trans C18:2, C18:3) and absorbed amounts (g/d) of each AA (Arg, His, Ile, Leu, Lys, Met, Phe, Thr, Trp, Val) were calculated and used as candidate variables in the models. A multi-model inference method was used to fit a large set of mixed models with study as the random effect, and the best models were selected based on Akaike's information criterion corrected for sample size and evaluated further. Observed milk fat concentration (MFC) ranged from 2.26 to 4.78%, and milk fat yield (MFY) ranged from 0.488 to 1.787 kg/d among studies. Dietary levels of forage, starch, and total FA (dry matter basis) averaged 50.8 ± 10.3% (mean ± standard deviation), 27.5 ± 7.0%, and 3.4 ± 1.3%, respectively. The MFC was positively correlated with dietary forage (0.294) and negatively associated with dietary starch (?0.286). The DI of C18:2 (g/d) was more negatively correlated with MFC (?0.313) than that of the other FA. The best variables for predicting MFC were days in milk, FA-free dry matter intake, forage, starch, DI of C18:2, DI of C18:3, and absorbed Met, His, and Trp. The best predictor variables for MFY were FA-free dry matter intake, days in milk, absorbed Met and Ile, and intakes of digested C16:0 and C18:3. This model had a root mean square error of 14.1% and concordance correlation coefficient of 0.81. Surprisingly, DI of C18:3 was positively related to milk fat, and this relationship was consistently observed among models. The models developed can be used as a practical tool for predicting milk fat of dairy cows, while recognizing that additional factors are likely to also affect fat yield.     

Effect of dietary protein concentration and degradability on response to rumen-protected methionine in lactating dairy cows

An incomplete 8 × 8 Latin square trial (4-wk periods; 12 wk total) using 32 multiparous and 16 primiparous Holstein cows was conducted to assess the production response to crude protein (CP), digestible rumen-undegraded protein (RUP), and rumen-protected Met (RPM; fed as Mepron; Degussa Corp., Kennesaw, GA). Diets contained [dry matter (DM) basis] 21% alfalfa silage, 34% corn silage, 22 to 26% high-moisture corn, 10 to 14% soybean meal, 4% soyhulls, 2% added fat, 1.3% minerals and vitamins, and 27 to 28% neutral detergent fiber. Treatments were a 2 × 2 × 2 factorial arrangement of the following main effects: 15.8 or 17.1% dietary CP, with or without supplemental rumen-undegraded protein (RUP) from expeller soybean meal, and 0 or 9 g of RPM/d. None of the 2- or 3-way interactions was significant. Higher dietary CP increased DM intake 1.1 kg/d and yield of milk 1.7 kg/d, 3.5% fat-corrected milk (FCM) 2.2 kg/d, fat 0.10 kg/d, and true protein 0.05 kg/d, and improved apparent N balance and DM and fiber digestibility. However, milk urea N and estimated urinary excretion of urea-N and total-N also increased, and apparent N efficiency (milk-N/N-intake) fell from 33 to 30% when cows consumed higher dietary CP. Positive effects of feeding more RUP were increased feed efficiency and milk fat content plus 1.8 kg/d greater FCM and 0.08 kg/d greater fat, but milk protein content was lower and milk urea N and urinary urea excretion were elevated. Supplementation with RPM increased DM intake 0.7 kg/d and FCM and fat yield by 1.4 and 0.06 kg/d, and tended to increase milk fat content and yield of milk and protein.