Essential amino acid profile of supplemental metabolizable protein affects mammary gland metabolism and whole-body glucose kinetics in dairy cattle

This study investigated mammary gland metabolism and whole-body (WB) rate of appearance (Ra) of glucose in dairy cattle in response to a constant supplemental level of metabolizable protein (MP) composed of different essential AA (EAA) profiles. Five multiparous rumen-fistulated Holstein-Friesian dairy cows (2.8 ± 0.4 lactations; 81 ± 11 d in milk; mean ± standard deviation) were abomasally infused according to a 5 × 5 Latin square design with saline (SAL) or 562 g/d of EAA delivered in different profiles where individual AA content corresponded to their relative content in casein. The profiles consisted of (1) a complete EAA 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). A total mixed ration (58% corn silage, 16% alfalfa hay, and 26% concentrate on a 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 on an individual cow basis. Each experimental period consisted of 5 d of continuous abomasal infusion followed by 2 d of no infusion. Arterial and venous blood samples were collected on d 4 of each period for determination of mammary gland AA and glucose metabolism. On d 5 of each period, D-[U-¹³C]glucose (13 mmol priming dose; continuous 3.5 mmol/h for 520 min) was infused into a jugular vein and arterial blood samples were collected before and during infusion to determine WB Ra of glucose. Milk protein yield did not differ between EAAC, GR1+ILV, and GR1+ALT, or between SAL and ILV, and increased over SAL and ILV with EAAC and GR1+ILV. Mammary plasma flow increased with ILV infusion compared with EAAC and GR1+ILV. Infusion of EAAC tended to increase mammary gland net uptake of total EAA and decreased the mammary uptake to milk protein output ratio (U:O) of non-EAA compared with SAL. Infusion of ILV increased mammary net uptake and U:O of Ile, Leu, and Val markedly over all treatments. The U:O of total Ile, Leu, and Val increased numerically (25%) with GR1+ILV infusion compared with EAAC, and the U:O of total Arg, Lys, and Thr tended to decrease, primarily from decreased U:O of Lys. During GR1+ALT infusion, U:O of total Arg, Lys, and Thr was greater than that during EAAC infusion, whereas U:O of Ile, Leu, and Val did not differ from EAAC. Glucose WB Ra increased 16% with GR1+ALT over SAL, and increased numerically 8 and 12% over SAL with EAAC and GR1+ILV, respectively. The average proportion of lactose yield relative to glucose WB Ra did not differ across treatments and averaged 0.53. On average, 28% of milk galactose arose from nonglucose precursors, regardless of treatment. In conclusion, intramammary catabolism of group 2 AA increased to support milk component synthesis when the EAA profile of MP was incomplete with respect to casein. Further, WB and mammary gland glucose metabolism was flexible in support of milk component synthesis, regardless of absorptive EAA profile.     

Effects of peripartum propylene glycol supplementation on nitrogen metabolism, body composition, and gene expression for the major protein degradation pathways in skeletal muscle in dairy cows

Early-lactating dairy cows mobilize body protein to provide amino acids that are directed toward gluconeogenesis and milk protein synthesis. Propylene glycol (PG) is a precursor of ruminal propionate, and feeding PG has been reported to improve energy supply by increasing blood glucose. Our hypothesis was that feeding PG could spare body protein by providing an alternative source of carbon for gluconeogenesis. The major objectives of this study were 1) to delineate the effects of pre- and postpartum PG supplementation in transition dairy cows on whole-body nitrogen balance, urinary 3-methylhistidine (3-MH) excretion, body composition, and gene expression profiles for the major protein degradation pathways in skeletal muscle; and 2) to characterize the changes in body protein metabolism during the periparturient period. Sixteen pregnant cows (7 primiparous and 9 multiparous) were paired based on expected calving dates and then randomly assigned within each pair to either a basal diet (control) or basal diet plus 600 mL/d of PG. Diets were fed twice daily for ad libitum intake, and PG was fed in equal amounts as a top dress from d -7 to d 45. All measurements were conducted at 3 time intervals starting at d -14 ± 5, d 15, and d 38 relative to calving. Propylene glycol had no effect on whole-body N balance, urinary 3-MH excretion, or body composition. However, N balance was lower at d 15 and 38, compared with d -14. Urinary excretion of 3-MH was lower at d -14 than at d 15 and 38. Supplemental PG had no effect on body weight (BW) and all components of empty BW. On average, cows fed both diets mobilized 19 kg of body fat and 14 kg of body protein between d -14 and d 38. Supplemental PG had no effect on mRNA abundance in skeletal muscle for m-calpain, and the 14-kDa ubiquitin-carrier protein E2 (14-kDa E2) and proteasome 26S subunit-ATPase components of the ubiquitin-mediated proteolytic pathway; however, PG supplementation downregulated mRNA expression for μ-calpain at d 15, and tended to downregulate mRNA expression for ubiquitin at d 15 and 38. Relative to calving, mRNA abundance for m- and μ-calpain, ubiquitin, and 14-kDa E2 were greater at d 15 compared with d -14 and d 38. In summary, these results indicate that transitional effects on whole-body metabolism and gene expression for the Ca²⁺-dependent and ubiquitin-mediated proteolytic pathways in skeletal muscle were more pronounced than those elicited by PG supplementation.     

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 silage protein degradability and fermentation acids on metabolizable protein concentration: A meta-analysis of dairy cow production experiments

A meta-analysis was conducted using data from dairy cow production studies to evaluate silage metabolizable protein (MP) concentrations. The data consisted of 397 treatment means in 130 comparisons, in which the effects of silage factors (e.g., date of harvest, wilting, silage additives) were investigated. Within a comparison, a fixed amount of the same concentrate was fed. A prerequisite of data to be included in the analysis was that silage dry matter (DM), crude protein (CP), ammonia N, lactic acid (LA), and total acid (TA) concentrations and digestibility were determined. A smaller data set (n = 248) comprised studies in which silage water-soluble N concentration was also analyzed. The supply of MP was estimated as amino acids absorbed from the small intestine using a model with constant values for ruminal effective protein degradability (EPD) and intestinal digestibility of rumen undegraded protein. Microbial protein was calculated on the basis of digestible carbohydrates and rumen degradable protein (RDP). Alternative models were used to estimate microbial protein formation, assuming the energy values of RDP and TA to be equivalent to 1.00, 0.75, 0.50, 0.25, and 0 times that of digestible carbohydrates. Because EPD values are seldom determined in production trials, they were derived using empirical models that estimate them from other feed components. The goodness of fit of models was compared on the basis of root mean squared error (RMSE) of milk protein yield (MPY) predicted from MP supply (adjusted for random study effect) and Akaike's information criterion. Metabolizable protein supply calculated from basal assumptions predicted MPY precisely within a study (RMSE = 16.2 g/d). Variable contribution of RDP to the energy supply for microbial synthesis influenced the precision of MPY prediction very little, but RMSE for MPY increased markedly when the energy supply of rumen microbes was corrected for TA concentration. Using predicted rather than constant EPD values also increased RMSE of MPY prediction. These observations do not mean that the supply of MP from undegraded feed protein is constant. However, it suggests that our current methods overestimate the range in EPD values and that the techniques have so many inherent technical problems that they can mask the true differences between the feeds. Including new elements in feed protein evaluation models may not improve the precision of production response predictions unless the consequent effects on the supply of other nutrients are taken into account.     

Lactational performance and plasma and muscle amino acid concentrations in dairy cows fed diets supplying 2 levels of digestible histidine and metabolizable protein

The objective of this experiment was to investigate the effect of dietary levels of digestible histidine (dHis) and MP on lactational performance and plasma and muscle concentrations of free AA in dairy cows. A randomized block design experiment was conducted with 48 Holstein cows, including 20 primiparous, averaging (±SD) 103 ± 22 d in milk and 45 ± 9 kg/d milk yield at the beginning of the experiment. A 2-wk covariate period preceded 12 experimental wk, of which 10 wk were for data and sample collection. Experimental treatments were (1) MP-adequate (MPA) diet with 2.1% dHis of MP (MPA2.1), (2) MPA with 3.0% dHis (MPA3.0), (3) MP-deficient (MPD) diet with 2.1% dHis (MPD2.1), and (4) MPD with 3.0% dHis (MPD3.0). Actual dHis supply was estimated at 64, 97, 57, and 88 g/d, respectively. Diets supplied MP at 110% (MPA) and 96% (MPD) of NRC 2001 dairy model requirements calculated based on DMI and production data during the experiment. Dry matter intake and milk yield data were collected daily, milk samples for composition and blood samples for AA analysis were collected every other week, and muscle biopsies at the end of covariate period, and during wk 12 of the experiment. The overall DMI was not affected by dHis or MP level. Milk yield tended to be increased by 3.0% dHis compared with 2.1% dHis. Milk true protein concentration and yield were not affected by treatments, whereas milk urea nitrogen concentration was lower for MPD versus the MPA diet. Milk fat concentration was lower for MPD versus MPA. There was a MP × dHis interaction for milk fat yield and energy-corrected milk; milk fat was lower for MPD3.0 versus MPD2.1, but similar for cows fed the MPA diet regardless of dHis level whereas energy-corrected milk was greater for MPA3.0 versus MPA2.1 but tended to be lower for MPD3.0 versus MPD2.1. Plasma His concentration was greater for cows fed dHis3.0, and concentration of sum of essential AA was greater, whereas carnosine, 1-Methyl-His and 3-Methyl-His concentrations were lower for cows fed MPA versus MPD diet. Muscle concentration of His was greater for cows fed dHis3.0 treatment. The apparent efficiency of His utilization was increased at lower MP and His levels. Overall, cows fed a corn silage-based diet supplying MP at 110% of NRC (2001) requirements tended to have increased ECM yield and similar milk protein yield to cows fed a diet supplying MP at 96% of requirements. Supplying dHis at 3.0% of MP (or 86 and 96 g/d, for MPD3.0 and MPA3.0, respectively) tended to increase milk yield and increased plasma and muscle concentrations of His but had minor or no effects on other production variables in dairy cows.     

Varying type of forage, concentration of metabolizable protein, and source of carbohydrate affects nutrient digestibility and production by dairy cows

The effects of forage source, concentration of metabolizable protein (MP), type of carbohydrate, and their interactions on nutrient digestibility and production were evaluated using a central composite treatment design. All diets (dry basis) contained 50% forage that ranged from 25:75 to 75:25 alfalfa silage:corn silage. Rumen-degradable protein comprised 10.7% of the dry matter (DM) in all diets, but undegradable protein ranged from 4.1 to 7.1%, resulting in dietary MP concentrations of 8.8 to 12.0% of the DM. Dietary starch ranged from 22 to 30% of the DM with a concomitant decrease in neutral detergent fiber concentrations. A total of 15 diets were fed to 36 Holstein cows grouped in 6 blocks. Each block consisted of three 21-d periods, and each cow was assigned a unique sequence of 3 diets, resulting in 108 observations. Milk production and composition, feed intake, and digestibility of major nutrients (via total collection of feces and urine) were measured. Few significant interactions between main effects were observed. Starch concentration had only minor effects on digestibility and production. Replacing corn silage with alfalfa decreased digestibility of N but increased digestibility of neutral detergent fiber. Increasing the concentration of MP increased N digestibility. The concentration (Mcal/kg) of dietary digestible energy (DE) increased linearly as starch concentration increased (very small effect) and was affected by a forage by MP interaction. At low MP, high alfalfa reduced DE concentration, but at high MP, increasing alfalfa increased DE concentration. Increasing alfalfa increased DM and DE intakes, which increased yields of energy-corrected milk, protein, and fat. Increasing MP increased yields of energy-corrected milk and protein. The response in milk protein to changes in MP was much less than predicted using the National Research Council (2001) model.     

Varying forage type, metabolizable protein concentration, and carbohydrate source affects manure excretion, manure ammonia, and nitrogen metabolism of dairy cows

Effects of forage source, concentration of metabolizable protein (MP), and type of carbohydrate on manure excretion by dairy cows and production of ammonia from that manure were evaluated using a central composite experimental design. All diets (dry basis) contained 50% forage that ranged from 25:75 to 75:25 alfalfa silage:corn silage. Diets contained 10.7% rumendegradable protein with variable concentrations of undegradable protein so that dietary MP ranged from 8.8 to 12%. Starch concentration ranged from 22 to 30% with a concomitant decrease in neutral detergent fiber. A total of 15 diets were fed to 36 Holstein cows grouped in 6 blocks. Each block was a replicated 3 × 3 Latin square resulting in 108 observations. Manure output (urine and feces) was measured using total collection, and fresh feces and urine were combined into slurries and incubated for 48 h to measure NH₃-N production. Feces, urine, and manure output averaged 50.5, 29.5, and 80.1 kg/d, respectively. Manure output increased with increasing dry matter intake (∼3.5 kg of manure/kg of dry matter intake), increased concentrations of alfalfa (mostly via changes in urine output), and decreased concentrations of starch (mostly via changes in fecal output). The amount of NH₃-N produced per gram of manure decreased with increasing alfalfa because excreted N shifted from urine to feces. Increasing MP increased NH₃-N produced per gram of manure mainly because of increased urinary N, but increased fecal N also contributed to the manure NH₃. Manure NH₃-N production per cow (accounts for effects on manure production and NH₃-N produced per unit of manure) was least and milk protein yields were maximal for diets with high alfalfa (75% of the forage), moderate MP (11% of diet dry matter), and high starch (30% of diet dry matter).     

Effects of jugular-infused lysine, methionine, and branched-chain amino acids on milk protein synthesis in high-producing dairy cows

In addition to lysine and methionine, current ration-balancing programs suggest that branched-chain amino acid (BCAA) supply may also be limiting in dairy cows. The objective of this study was to investigate whether BCAA, leucine, isoleucine, and valine become limiting for milk protein synthesis when methionine and lysine supply were not limiting. Nine multiparous Holstein cows with an average milk production of 53.5 ± 7.1 kg/d were randomly assigned to 7-d continuous jugular infusions of saline (CTL), methionine and lysine (ML; 12 g and 21 g/d, respectively), or ML plus leucine, isoleucine, and valine (ML+BCAA; 35 g, 15 g, and 15 g/d, respectively) in a 3 × 3 Latin square design with 3 infusion periods separated by 7-d noninfusion periods. The basal diet consisted of 40% corn silage, 14% alfalfa hay, and a concentrate mix, and respectively supplied lysine, methionine, isoleucine, leucine, and valine as 6.1, 1.8, 4.7, 8.9, and 5.3% of metabolizable protein. Dry matter intake (23.9 kg/d), milk yield (52.8 kg/d), fat content (2.55%), fat yield (1.33 kg/d), lactose content (4.77%), lactose yield (2.51 kg/d), and milk protein efficiency (0.38) were similar across treatments. Protein yield and protein content were not significantly different between ML (1.52 kg/d and 2.88%, respectively) and ML+BCAA (1.51 kg/d and 2.83%, respectively), but they were significantly greater than that of CTL (1.39 kg/d and 2.71%). Cows that received ML+BCAA had less milk urea nitrogen content (10.9 mg/dL) compared with milk of CTL cows (12.4 mg/dL) and ML cows (11.8 mg/dL). Whereas high-producing cows responded positively to methionine and lysine supplementation, no apparent benefits of BCAA supplementation in milk protein synthesis were found. Infusion of BCAA may have stimulated synthesis of other body proteins, probably muscle proteins, as evidenced by decreased milk urea nitrogen.     

Effect of source of rumen-degraded protein on production and ruminal metabolism in lactating dairy cows

Twenty-eight (8 with ruminal cannulas) lactating Holstein cows were assigned to seven 4 × 4 Latin squares in a 16-wk trial to study the effects on production and ruminal metabolism of feeding differing proportions of rumen-degraded protein (RDP) from soybean meal and urea. Diets contained [dry matter (DM) basis] 40% corn silage, 15% alfalfa silage, 28 to 30% high-moisture corn, plus varying levels of ground dry shelled corn, solvent- and lignosulfonate-treated soybean meal, and urea. Proportions of the soybean meals, urea, and dry corn were adjusted such that all diets contained 16.1% crude protein and 10.5% RDP, with urea providing 0, 1.2, 2.4, and 3.7% RDP (DM basis). As urea supplied greater proportions of RDP, there were linear decreases in DM intake, yield of milk, 3.5% fat-corrected milk, fat, protein, and solids-not-fat, and of weight gain. Milk contents of fat, protein, and solids-not-fat were not affected by source of RDP. Replacing soybean meal RDP with urea RDP resulted in several linear responses: increased excretion of urinary urea-N and concentration of milk urea-N, blood urea-N, and ruminal ammonia-N and decreased excretion of fecal N; there was also a trend for increased excretion of total urinary N. A linear increase in neutral detergent fiber (NDF) digestibility, probably due to digestion of NDF-N from lignosulfonate-treated soybean meal, was observed with greater urea intake. Omasal sampling revealed small but significant effects of N source on measured RDP supply, which averaged 11.0% (DM basis) across diets. Increasing the proportion of RDP from urea resulted in linear decrease in omasal flow of dietary nonammonia N (NAN) and microbial NAN and in microbial growth efficiency (microbial NAN/unit of organic matter truly digested in the rumen). These changes were paralleled by large linear reductions in omasal flows of essential, nonessential, and total amino acids. Overall, these results indicated that replacing soybean meal RDP with that from urea reduced yield of milk and milk components, largely because of depressed microbial protein formation in the rumen and that RDP from nonprotein-N sources was not as effective as RDP provided by true protein.     

Reducing metabolizable protein supply: Effects on milk production,blood metabolites,and health in early-lactation dairy cows

Our objective was to evaluate the effect of metabolizable protein (MP) supply on milk production, blood metabolites, and health in dairy cows during early lactation. Three experimental diets were formulated to contain 114, 107, 101 g of MP/kg of dry matter (DM; 114MP, 107MP, and 101MP, respectively) with crude protein contents of 17.0, 16.2, and 15.3% of DM, respectively. One hundred multiparous Holstein cows were fed 1 of these 3 diets during wk 1 to 3 and wk 4 to 13 of lactation in one of the following sequences: (1) 114MP and 107MP (114MP/107MP), (2) 114MP and 101MP (114MP/101MP), or (3) 101MP and 101MP (101MP/101MP). During wk 1 to 3, the 114MP and 101MP treatments were 20 and 27% deficient in estimated MP, respectively. From wk 4 to 13, the 114MP/107MP, 114MP/101MP, and 101MP/101MP treatments were 8, 12, and 13% deficient in estimated MP, respectively. Data were analyzed separately for wk 1 to 3, 4 to 13, and 1 to 13. Dry matter intake and energy-corrected milk (ECM) yield were not affected by treatment during wk 4 to 13 or wk 1 to 13; however, ECM yield decreased for 101MP versus 114MP from wk 1 to 3. Similarly, feed efficiency was not affected by treatment from wk 4 to 13 or wk 1 to 13, and was reduced with 101MP versus 114MP during wk 1 to 3. Milk N efficiency tended to increase for 101MP versus 114MP for wk 1 to 3 and increased with 101MP/101MP and 114MP/101MP relative to 114MP/107MP during wk 4 to 13 and wk 1 to 13. Treatment had no influence on yields and concentrations of milk components from wk 4 to 13 or wk 1 to 13; however, compared with 114MP, feeding 101MP tended to decrease milk fat yield and decreased yields of milk true protein and lactose for wk 1 to 3. Both milk and blood urea N concentrations decreased for 101MP/101MP and 114MP/101MP relative to 114MP/107MP during wk 4 to 13 and wk 1 to 13, and were reduced with feeding 101MP versus 114MP from wk 1 to 3. Treatment had no effect on the incidence of diseases in cows throughout the study. Serum concentrations of total fatty acids, albumin, and aspartate aminotransferase did not differ between 101MP and 114MP; however, serum β-hydroxybutyrate concentration was lower in cows receiving 101MP during the first 3 wk of lactation. Compared with 114MP, feeding 101MP during wk 1 to 3 increased plasma concentrations of creatinine and 3-methylhistidine (3-MHis) but did not change the ratio of plasma 3-MHis to creatinine. We found no differences in plasma creatinine or the ratio of 3-MHis-to-creatinine among treatments from wk 4 to 13; however, 101MP/101MP and 114MP/101MP had elevated plasma 3-MHis compared with 114MP/107MP. Treatment had no effect on body weight and body condition score over the duration of the study. Collectively, despite reduced milk production for the first 3 wk of lactation, feeding the 101MP/101MP treatment sustained lactational performance and improved milk N efficiency without negatively affecting the frequency of diseases in dairy cows during the first 13 wk postpartum.     

Effects of metabolizable protein supply and amino acid supplementation on nitrogen utilization, milk production, and ammonia emissions from manure in dairy cows

Two experiments were conducted with the objective of investigating the effects of rumen-protected methionine (RPMet) supplementation of metabolizable protein (MP)-deficient or MP-adequate but Met-deficient diets on dairy cow performance. Experiment (Exp.) 1 utilized 36 Holstein dairy cows blocked in 12 blocks of 3 cows each. Cows within block were assigned to one of the following dietary treatments: (1) MP-adequate diet [AMP; positive MP balance according to the National Research Council (2001) dairy model]; (2) an MP-deficient diet supplemented with 100g of rumen-protected Lys (RPLys)/cow per day (DMPL); and (3) DMPL supplemented with 24g of RPMet/cow per day (DMPLM). Experiment 2 utilized 120 Holstein cows assigned to 6 pens of 20 cows each. Pens (3 per treatment) were assigned to one of the following dietary treatments: (1) AMP diet supplemented with 76g of RPLys/cow per day (AMPL); and (2) AMPL (74g of RPLys/cow per day) supplemented with 24g of RPMet/cow per day (AMPLM). Each experiment lasted for 10wk (2-wk adaptation and 8-wk experimental periods) following a 2-wk covariate period (i.e., a total of 12wk). In Exp. 1, the MP-deficient diets decreased apparent total-tract nutrient digestibility but had no statistical effect on dry matter intake (DMI), milk yield, or milk fat percentage and yield. Compared with AMP, DMPL decreased milk protein content; both DMPL and DMPLM diets decreased milk protein yield. Urinary N losses and milk urea-N concentration were decreased by the MP-deficient diets compared with AMP. The ammonia emitting potential of manure from the MP-deficient diets was decreased by about 37% compared with that of AMP manure. Plasma Lys and Met concentrations were not affected by treatment, but concentrations of His, Thr, and Val were lower for the MP-deficient diets compared with AMP. In Exp. 2, the AMPLM diet had lower milk yield than AMPL due to numerically lower DMI; no other effects were observed in Exp. 2. In conclusion, feeding MP-deficient diets supplemented with RPLys and RPMet did not statistically decrease milk yield in dairy cows in Exp. 1. However, without RPMet supplementation, milk protein content was decreased compared with the MP-adequate diet. Other amino acids, possibly His, may limit milk production in MP-deficient, corn or corn silage-based diets. A summary of 97 individual cow data from trials in which MP-deficient diets were fed suggested the National Research Council (2001) model under-predicts milk yield in cows fed MP-deficient diets (MP balance of -20 to -666g/d) in a linear manner: milk yield under-prediction [National Research Council (2001) MP-allowable milk yield, kg/d - actual milk yield, kg/d] = 0.0991 (±0.0905) + 0.0230 (±0.0003) × MP balance, g/d (R(2)=0.99).     

Effects of glucagon infusions on protein and amino acid composition of milk from dairy cows

Changing the composition of milk proteins and AA affects the nutritional and physical properties of dairy products. Intravenous infusions of glucagon decreases milk protein production and concentration by promoting the use of gluconeogenic blood AA for hepatic glucose synthesis. Little is known about how the diversion of AA to gluconeogenesis affects the composition of milk proteins and AA. The objective was to quantify changes in composition of milk protein and AA in response to i.v. glucagon infusions. Three separate experiments were used: 1) 8 Holstein cows were fed ad libitum and infused with glucagon at 10 mg/d for 14 d, 2) 7 Holstein cows were feed restricted and infused with glucagon at 10 mg/d for 14 d, and 3) 4 Brown Swiss cows were infused with glucagon at 5 and 10 mg/d for 2 d each. Milk and milk component yields and milk protein and amino acid composition of samples, collected with blood samples at the first and last day of the glucagon infusion period, were compared with those collected 1 d before and after the glucagon infusion period. Glucagon infusions decreased milk protein production and concentration in each experiment by at least 0.2 ± 0.05 kg/d and 4 ± 0.4 g/L, respectively. The decrease was accompanied by changes in milk protein composition, the most consistent being an increase in κ-casein (1.68 ± 0.27%). Overall, glucagon infusions resulted in higher proportions of κ-casein and α_S2-casein (1.34 ± 0.51%) and smaller proportions of α_S1-casein (−3.83 ± 1.75%) and α-lactalbumin (−0.91 ± 0.32%). Glucagon had little impact on milk AA composition except an increase in glycine (0.26 ± 0.11%). The results suggest that milk protein synthesis is regulated by many factors including AA and glucose availability.     

Predicting milk protein responses and the requirement of metabolizable protein by lactating dairy cows

The objective of this study was to develop a modeling framework to predict milk protein yield responses to varying metabolizable protein (MP) supplies and to determine the requirement of MP by lactating dairy cows. The logistic curve was used to model milk protein yield while accounting for a variable efficiency of MP utilization and between-study variability. Models were developed with databases from 2 recently published meta-analyses and based on either total MP supply or MP supply available for milk production. All models provided reasonable fit to data, with root mean square prediction error ranging from 18 to 20% of the average milk protein yield. The estimated horizontal asymptotes were 1.17 (posterior SD = 0.02) and 1.55 (posterior SD = 0.06) in the 2 databases, suggesting that the limiting milk protein yield, as MP supply increases, converges to 1.17 or 1.55 kg/d in the environments determined by the 2 databases. The observed efficiencies ranged from 0.75 to 0.18 when total MP supply was used as the denominator and above 1 to 0.24 when the MP supply available for milk production was used as the denominator. The predicted efficiencies were in good agreement with the data, decreasing nonlinearly with the MP supply. The MP requirement was calculated with a function constructed with the inverse of the logistic model and modified at regions of maximum marginal efficiency and minimum second derivative. This strategy assumes that the MP solution, or the MP needed to predict a given protein yield in the fitted logistic curve, determines the MP requirement for maintenance and lactation. Requirements calculated with the independent variable as total MP supply refer to the total requirement of maintenance plus lactation, whereas the requirement from models based on MP supply available for milk production are referent to the MP required only for lactation. The requirements were, on average, slightly smaller than the ones predicted by the current Northern American feeding system for dairy cows at lower protein yields and greater than currently recommended at high yields.     

Effect of protein source on amino acid supply,milk production,and metabolism of plasma nutrients in dairy cows fed grass silage

This study conducted according to a 4 x 4 Latin square with 28 d periods and four ruminally cannulated Finnish Ayrshire cows investigated the effect of protein supplements differing in amino acid (AA) profile and rumen undegradable protein content on postruminal AA supply and milk production. Mammary metabolism of plasma AA and other nutrients were also studied. The basal diet (Control; 13.4% crude protein) consisted of grass silage and barley in a ratio of 55:45 on a dry matter basis. The other three isonitrogenous diets (17.0% crude protein) were control + fishmeal (FM), control + soybean meal (SBM), and control + corn gluten meal (CGM). The protein supplements replaced portions of dry matter of the control diet maintaining the silage to barley ratio constant for all diets. Dry matter intake was limited to 95% of the preexperimental ad libitum intake and was similar (mean 19.8 kg/d dry matter) across the diets. Protein supplements increased milk, lactose, and protein yields but did not affect yields of energy-corrected milk or milk fat. Milk protein yield response was numerically lowest for diet SBM. Protein supplements increased milk protein concentration but decreased milk fat and lactose concentrations. Microbial protein synthesis and rumen fermentation parameters were similar across the diets, except for an increased rumen ammonia concentration for diets supplemented with protein feeds. Protein supplements increased N intake, ruminal organic matter and N, and total tract organic matter, N, and neutral detergent fiber digestibilities. Protein supplements also increased N and AA flows into the omasum, with SBM giving the lowest and CGM the highest flows. This was associated with an unchanged microbial N flow and a higher undegraded dietary N flow. The omasal flows of individual AA reflected differences in total N flow and AA profile of the experimental diets. Differences in AA flows did not always reflect plasma AA concentrations. The results indicated that AA supply of dairy cows fed a grass silage-cereal diet can be manipulated using protein supplements differing in ruminal protein degradability and AA profile. Lower milk production response to SBM than that to FM and CGM appeared to be related mainly to lower N and AA supplies arising from a high ruminal protein degradability of SBM. Histidine appeared to be the first limiting AA for milk protein synthesis on the control diet. Mammary gland may regulate AA uptake according to requirements.     

Replacing soybean meal with okara meal: Effects on production,milk fatty acid and plasma amino acid profile,and nutrient utilization in dairy cows

Okara meal is a byproduct from the production of soymilk and tofu and can potentially replace soybean meal (SBM) in dairy diets due to its high crude protein (CP) concentration and residual fat. The objective of this study was to investigate the effects of replacing SBM with okara meal on feed intake, yields of milk and milk components, milk fatty acid (FA) profile, nutrient utilization, and plasma AA concentration in lactating dairy cows. Twelve multiparous (65 ± 33 d in milk) and 8 primiparous (100 ± 35 d in milk) organically certified Jersey cows were paired by parity or days in milk, and within pair, randomly assigned to treatments in a crossover design with 21-d periods (14 d for diet adaptation and 7 d for data and sample collection). Diets were fed as total mixed ration formulated to be isonitrogenous and isofibrous and contained (dry matter basis) 50% mixed, mostly grass baleage, 2% sugarcane liquid molasses, 2% minerals-vitamins premix, and either (1) 8.1% SBM, 10% soyhulls, and 27.9% ground corn (CTRL); or (2) 15% okara meal, 8% soyhulls, and 23% ground corn (OKR). Dietary CP, ash-free neutral detergent fiber, and total FA averaged 15.4, 35.3, and 3.08% for CTRL and 15.9%, 36.3%, and 3.74% for OKR, respectively. Substitution of SBM with okara meal did not alter dry matter intake but increased intakes of CP and ash-free neutral detergent fiber. Additionally, no significant differences between treatments were observed for yields of milk and milk components, and concentrations of milk fat, lactose, and total solids. However, milk true protein concentration was lower in cows fed OKR (3.76%) versus CTRL (3.81%). Both milk urea N (8.51 vs. 9.47 mg/dL) and plasma urea N (16.9 vs. 17.8 mg/dL) concentrations decreased with OKR relative to the CTRL diet, respectively. Compared with CTRL, feeding OKR lowered the milk proportions of total odd-chain FA, de novo FA, and mixed FA and increased those of preformed FA, total n-6 FA, and total n-3 FA. The milk proportions of trans-10 18:1, trans-11 18:1, and cis-9,trans-11 18:2 were greater with feeding OKR versus the CTRL diet. The apparent total-tract digestibility of nutrients, urinary excretion of total purine derivatives (uric acid plus allantoin), and total N were not affected by treatments. Except for plasma Leu, which was lower in OKR compared with the CTRL diet, no other significant changes in the plasma concentrations of AA were observed. The plasma concentration of carnosine was lowest in cows receiving the OKR diet. Overall, our results revealed that okara meal can completely replace SBM without negatively affecting production and nutrient digestibility in early- to mid-lactation Jersey cows. Further research is needed to assess the economic feasibility of including okara meal in dairy diets, as well as the amount of okara meal that maximizes yields of milk and milk components in dairy cows in different stages of lactation.     

Effects of different protein supplements on omasal nutrient flow and microbial protein synthesis in lactating dairy cows

Eight ruminally cannulated Holstein cows that were part of a larger lactation trial were used in 2 replicated 4 × 4 Latin squares to quantify effects of supplementing protein as urea, solvent soybean meal (SSBM), cottonseed meal (CSM), or canola meal (CM) on omasal nutrient flows and microbial protein synthesis. All diets contained (% of dry matter) 21% alfalfa silage and 35% corn silage plus 1) 2% urea plus 41% high-moisture shelled corn (HMSC), 2) 12% SSBM plus 31% HMSC, 3) 14% CSM plus 29% HMSC, or 4) 16% CM plus 27% HMSC. Crude protein was equal across diets, averaging 16.6%. The CSM diet supplied the least rumen-degraded protein and the most rumen-undegraded protein. Microbial nonammonia N flow was similar among the true protein supplements but was 14% lower in cows fed urea. In vivo ruminal passage rate, degradation rate, and estimated escape for the 3 true proteins were, respectively, 0.044/h, 0.105/h, and 29% for SSBM; 0.051/h, 0.050/h, and 51% for CSM; and 0.039/h, 0.081/h, and 34% for CM. This indicated that CSM protein was less degraded because of both a faster passage rate and slower degradation rate. Omasal flow of individual AA, branched-chain AA, essential AA, nonessential AA, and total AA all were lower in cows fed urea compared with one of the true protein supplements. Among the 3 diets supplemented with true protein, omasal flow of Arg was greatest on CSM, and omasal flow of His was greatest on CSM, intermediate on CM, and lowest on SSBM. Lower flows of AA and microbial nonammonia N explained lower yields of milk yield and milk components observed on the urea diet in the companion lactation trial. These results clearly showed that supplementation with true protein was necessary to obtain sufficient microbial protein and rumen-undegraded protein to meet the metabolizable AA requirements of high-producing dairy cows.     

Changes in skeletal muscle thickness and echogenicity and plasma creatinine concentration as indicators of protein and intramuscular fat mobilization in periparturient dairy cows

High-producing dairy cows experience a state of negative energy balance in the periparturient period that is partially addressed by increasing the rate of fat and protein mobilization. Previous studies have focused on the rate of fat mobilization, and consequently the rate of protein mobilization has not been well characterized. The objective of this study was therefore to determine the change in indicators of muscle mass during early lactation using ultrasonographic measurement of muscle thickness and changes in plasma creatinine concentration. The maximum thickness of the gluteus medius and longissimus dorsi muscles of 106 Holstein cows (34 primiparous, 72 multiparous) was determined ultrasonographically on d ?3, 0, 3, 7, 14, 21, and 28 relative to the day of parturition. Plasma creatinine concentration was measured periodically during the same period. Mixed models analysis and Passing-Bablok regression were used to analyze the data. Gluteus medius thickness, longissimus dorsi loin thickness (LDLT), and longissimus dorsi thoracic thickness (LDTT) were decreased at 28 d postpartum compared with d 3 antepartum. Plasma creatinine concentration was weakly associated with gluteus medius thickness, LDLT, and LDTT (Spearman's rho = 0.31, 0.39, and 0.32, respectively). Plasma creatinine concentration in primiparous and multiparous cows at 28 d postpartum decreased by 0.24 and 0.30 mg/dL, respectively, compared with values 3 d antepartum. We concluded that ultrasonographic measurement of LDLT and LDTT and change in plasma creatinine concentration may provide practical methods for monitoring the rate of protein mobilization in periparturient dairy cows. Ultrasonographic examination of LDLT and LDTT therefore complements ultrasonographic measurement of backfat thickness and may be useful in the evaluation of energy reserve mobilization in periparturient dairy cows.     

Effects of body condition,monensin, and essential oils on ruminal lipopolysaccharide concentration,inflammatory markers,and endoplasmatic reticulum stress of transition dairy cows

Evidence exists that dairy cows experience inflammatory-like phenomena in the transition period. Rumen health and alterations in metabolic processes and gene networks in the liver as the central metabolic organ might be key factors for cows’ health and productivity in early lactation. This study made use of an animal model to generate experimental groups with different manifestations of postpartal fat mobilization and ketogenesis. In total, 60 German Holstein cows were allocated 6 wk antepartum to 3 high-body condition score (BCS) groups (BCS 3.95) and 1 low-BCS group (LC; BCS 2.77). High-BCS cows were fed an antepartal forage-to-concentrate ratio of 40:60 on dry matter basis, in contrast to 80:20 in the LC group, and received a monensin controlled-release capsule (HC/MO), a blend of essential oils (HC/EO), or formed a control group (HC). We evaluated serum haptoglobin, kynurenine, tryptophan, ruminal lipopolysaccharide concentration and mRNA abundance of nuclear factor kappa B (NF-κB), nuclear factor E2-related factor 2 (Nrf2), and endoplasmatic reticulum stress-induced unfolded protein response (UPR) target genes in liver biopsy samples from d ?42 until +56 relative to calving. Nearly all parameters were highly dependent on time, with greatest variation near calving. The ruminal lipopolysaccharide concentration and evaluated target genes were not generally influenced by antepartal BCS and feeding management. The kynurenine-to-tryptophan ratio was higher in LC than in HC/MO treatment on d 7. Ruminal lipopolysaccharide concentration was higher in HC/MO than in the HC group, but not increased in HC/EO group. Abundance of UPR target gene X-box binding protein 1 was higher in HC/MO than in HC/EO group on d 7. Hepatic mRNA abundance of Nrf2 target gene glutathione peroxidase 3 was higher, whereas expression of NF-κB target gene haptoglobin tended to be higher in LC than in HC/EO cows. The HC/MO cows showed the most prominent increase in the abundance of glutathione peroxidase 3 and haptoglobin after calving in comparison to antepartal values. Results indicate the presence of inflammatory-like phenomena near calving. Simultaneously, alterations in UPR and Nrf2 target genes with antioxidative properties and haptoglobin occurred, being most prominent in LC and HC/MO group.     

Effect of level of metabolizable protein on splanchnic flux of amino acids in lactating dairy cows

The response of splanchnic tissue metabolism to different levels of metabolizable protein (MP) was measured in 6 catheterized multiparous lactating Holstein cows. Three diets, balanced to provide similar energy intakes and increasing amounts of MP (g/d)-1922 (low), 2264 (medium), and 2517 (high)-were fed during 21-d experimental periods according to a replicated Latin square. On d 18, 19, or 20, six hourly blood samples were collected simultaneously from the portal and hepatic veins plus an artery to determine net fluxes of nutrients across the portal-drained viscera and the liver. Yields of milk and protein increased, as did urinary N excretion with increasing MP. Portal absorption of essential amino acids (EAA) increased linearly with increasing MP supply, as did liver removal of His, Met, and Phe. In contrast, liver removal of the branched-chain AA (BCAA) and lysine was unaffected by diets. With increasing MP, the ratio of milk output to postliver supply of BCAA, Thr, and Lys decreased linearly, indicating oxidation of these AA in the peripheral tissues. Concomitant to a decreased catabolism of EAA in the liver (His, Met, Phe, and Thr) and/or in peripheral tissues (BCAA, Lys, and Thr), the efficiency of transfer of absorbed EAA into milk protein decreases markedly as protein supply increases. The efficiency of transfer of absorbed AA into milk also varies greatly between AA. These 2 important factors should be taken into account when building predictive schemes for milk protein output.