Metabolizable Protein Balance Research Articles

364 The Effects of Replacing Dried Distillers Grains with Solubles with a Heat-Treated Soybean Meal in Forage-Based Growing Cattle Diets

Abstract This study aimed to evaluate the effects of increased concentrations of metabolizable protein and lysine from heat-treated soybean meal (TSBM) in forage-based growing cattle diets predicted to have positive metabolizable protein balance. Seventy Angus-based steers (298 ± 16 kg initial BW) were utilized in an 85-d randomized generalized block design with animal as the experimental unit. Dietary treatments were arranged to replace a proportion of the basal protein source, DDGS included at 16% dietary dry matter (DM), with TSBM at rates of 0% (TSBM0), 4% (TSBM4), 8% (TSBM8), and 12% (TSBM12) dietary DM. Steers were provided with ad libitum access to feed and water in a monoslope barn with dry-lot access. The basal diets contained (DM basis) 44% corn silage, 37% oat hay, and 3% supplement. Steers were blocked by BW into light, medium, and heavy blocks. Daily feed disappearance was measured using an automated feed system (Insentec Roughage Intake Control, Hokofarm B.V., Marknesse, The Netherlands), with disappearance assumed to be intake. Steers were individually weighed every 28 d to calculate average daily gain (ADG) throughout the experiment. Growth performance and feed intake were analyzed utilizing the MIXED procedure of SAS with fixed effects of body weight block and TSBM inclusion level. Treatments were evaluated when the F test was significant (P ≤ 0.05). There were no differences (P ≥ 0.16) for ending BW, ADG, dry matter intake (DMI), or gain:feed (G:F) among treatments (Table 1). Therefore, in growing diets predicted to be sufficient in metabolizable protein, replacing distillers grains with a source of metabolizable protein and amino acids provided by heat-treated soybean meal did not affect growth performance.

Effects of dietary protein concentration and degradability on performance, carcass characteristics, net energy utilization, and metabolizable protein balance of finishing beef heifers receiving 0 or 400 mg of ractopamine hydrochloride

This study evaluated the effects of feeding beef cattle finishing diets with greater than 14.0% dietary CP and with or without ractopamine hydrochloride (RH) on growth performance, carcass merit, net energy utilization, and metabolizable protein balance. Heifers (n = 525) were assigned to 48 pens in a generalized complete block design, and pens of cattle were assigned randomly to 0 or 400 mg of RH/animal per day and 3 CP treatments in a 2 × 3 factorial arrangement (n = 8 pens/treatment) fed for 35 d before slaughter. Dietary protein treatments were steamflaked corn-based diets containing 13.9% CP, 8.9% RDP, and 5.1% RUP (CON); 20.9% CP, 14.4% RDP, and 6.5% RUP (High RDP); or 20.9% CP, 9.7% RDP, and 11.2% RUP (High RUP) on a DM basis. No RH × CP interactions were observed. Final BW, ADG, water disappearance, DMI, and G:F were not different among CP treatments. Dressing percentage was greater for cattle fed High RDP than for those fed High RUP, but other carcass outcomes did not differ. The MP balance was greatest for High RUP, intermediate for High RDP, and least for CON. Cattle receiving 400 mg of RH had greater final BW, ADG, G:F, and hot carcass weight. The LM area was greater and KPH was less for 400 versus 0 mg of RH. Carcass-adjusted final BW, ADG, and G:F were greater for cattle consuming 400 mg of RH. Cattle fed 400 mg of RH had greater performance-adjusted and observed:expected NEm and NEg. The MP balance was less for 400 versus 0 mg of RH. Dietary CP requirements were greater for cattle fed 400 compared with 0 mg of RH but did not exceed the CP supplied by CON, High RDP, or High RUP. Feeding greater than 14.0% CP does not negatively affect performance or carcass characteristics of finishing cattle, and the absence of interactions between CP and RH suggests that RH does not increase CP requirements above those provided in a typical finishing cattle diet.

Balancing dairy cattle diets for rumen nitrogen and methionine or all essential amino acids relative to metabolizable energy

Improving the ability of diet formulation models to more accurately predict AA supply while appropriately describing requirements for lactating dairy cattle provides an opportunity to improve animal productivity, reduce feed costs, and reduce N intake. The goal of this study was to evaluate the sensitivity of a new version of the Cornell Net Carbohydrate and Protein System (CNCPS) to formulate diets for rumen N, Met, and all essential AA (EAA). Sixty-four high-producing dairy cattle were randomly assigned to 1 of the 4 following diets in a 14-wk longitudinal study: (1) limited metabolizable protein (MP), Met, and rumen N (Base), (2) adequate Met but limited MP and rumen N (Base + M), (3) adequate Met and rumen N, but limited MP (Base + MU), and (4) adequate MP, rumen N, and balanced for all EAA (Positive). All diets were balanced to exceed requirements for ME relative to maintenance and production, assuming a nonpregnant, 650-kg animal producing 40 kg of milk at 3.05% true protein and 4.0% fat. Dietary MP was 97.2, 97.5, 102.3, and 114.1 g/kg of dry matter intake for the Base, Base + M, Base + MU, and Positive diets, respectively. Differences were observed for dry matter intake and milk yield (24.1 to 24.7 and 39.4 to 41.1 kg/d, among treatments). Energy corrected milk, fat, and true protein yield were greater (2.9, 0.13, and 0.08 kg/d, respectively) in cows fed the Positive compared with the Base diet. Using the updated CNCPS, cattle fed the Base, Base + M, and Base + MU diets were predicted to have a negative MP balance (-231, -310, and -142 g/d, respectively), whereas cattle fed the Positive diet consumed 33 g of MP/d excess to ME supply. Bacterial growth was predicted to be depressed by 16 and 17% relative to adequate N supply for the Base and Base + M diets, respectively, which corresponded with the measured lower apparent total-tract NDF degradation. The study demonstrates that improvements in lactation performances can be achieved when rumen N and Met are properly supplied and further improved when EAA supply are balanced relative to requirements. Formulation using the revised CNCPS provided predictions for these diets, which were sensitive to changes in rumen N, Met, all EAA, and by extension MP supply.

Dietary starch level and rumen-protected methionine, lysine, and histidine: Effects on milk yield, nitrogen, and energy utilization in dairy cows fed diets low in metabolizable protein

Our objective was to investigate the interactions between starch level and rumen-protected Met, Lys, His (RP-MLH) on milk yield, plasma AA concentration, and nutrient utilization in dairy cows fed low metabolizable protein diets (mean = -119 g/d of metabolizable protein balance). Sixteen multiparous Holstein cows (138 ± 46 d in milk, 46 ± 6 kg/d in milk) were used in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Dietary starch level varied by replacing (dry matter basis) pelleted beet pulp and soyhulls with ground corn resulting in the following treatments: (1) 20% pelleted beet pulp and 10% soyhulls (reduced starch = RS), (2) RS plus RP-MLH (RS+AA), (3) 30% ground corn (high starch = HS), and (4) HS plus RP-MLH (HS+AA). Dietary starch concentrations averaged 12.3 and 34.4% for RS and HS basal diets, respectively. Diets were supplemented with RP-MLH products to supply digestible Met, Lys, and His. Compared with RS and RS+AA diets, HS and HS+AA diets increased yields of milk (37.9 vs. 40.1 kg/d) and milk protein (1.07 vs. 1.16 kg/d) and decreased dry matter intake (25.9 vs. 25.2 kg/d), milk urea N (12.6 vs. 11.0 mg/dL), and plasma urea N (13.3 vs. 11.6 mg/dL). Milk N efficiency was greater in cows fed the HS and HS+AA than RS and RS+AA diets (28.9 vs. 25%), and RP-MLH supplementation improved milk true protein concentration. Starch level × RP-MLH interactions were observed for plasma concentrations of Arg and Lys, with RP-MLH being more effective to increase plasma Arg (+16%) and Lys (+23%) when supplemented to the RS than the HS basal diet. Replacing pelleted beet pulp and soyhulls with ground corn lowered the plasma concentrations of all essential AA except Met and Thr. In addition, the plasma concentrations of His and Met increased with RP-MLH. The apparent total-tract digestibilities of neutral and acid detergent fiber were lower, and those of starch and ether extract greater in cows offered the HS and HS+AA diets than RS and RS+AA diets. Urinary excretion of urea N decreased by replacing pelleted beet pulp and soyhulls with ground corn. Enteric CH4 production, CH4 yield, and CH4 intensity all decreased in the HS and HS+AA versus RS and RS+AA diets. Diets did not affect the intakes of gross energy, metabolizable energy, and net energy of lactation. In contrast, digestible energy intake increased with feeding the RS and RS+AA diets, whereas CH4 energy decreased in cows fed the HS and HS+AA diets. Supplementation with RP-MLH had no effect on energy utilization variables. Overall, the lack of interactions between dietary starch level and RP-MLH supplementation on most variables measured herein showed that the effects of starch intake and RP-MLH were independent or additive.

Effect of protein level and methionine supplementation on dairy cows during the transition period

Cows experience a significant negative protein balance during the first 30 d of lactation. Given the functional effects of AA on health, especially in challenging periods such as calving, higher levels of protein and specific AA in the diet may act to improve health and feed intake. The response of dairy cows to 3 protein supplementation strategies during the transition period and through the first 45 d in milk was evaluated. The final data set had 39 Holstein cows blocked based on parity (primiparous vs. multiparous) and expected calving and randomly assigned within each block to one of 3 dietary treatments: low protein (LP), high protein (HP), or high protein plus rumen-protected methionine (HPM). Treatments were offered from d -18 ± 5 to 45 d relative to parturition. Pre- and postpartum diets were formulated for high metabolizable protein (MP) supply from soybean meal, and HP and HPM provided higher MP balance than LP. Preplanned contrasts were LP versus HP+HPM and HP versus HPM. Significance was declared at P ≤ 0.05 and trends at 0.05 <P ≤ 0.10. Cows fed HP and HPM had greater fry matter intake (DMI) prepartum than LP (+2 kg/d), and there was a trend for greater DMI with HPM than with HP (+1.6 kg/d). Body weight and condition score before and after calving did not differ among treatments. High protein (HP and HPM) tended to increase milk yield during the first 45 d of lactation (+1.75 kg/d), increased milk lactose content and urea-N in milk and plasma, tended to increase blood BHB 14 d postpartum, and tended to reduce milk/DMI compared with LP. Blood concentrations of calcium at calving and of glucose, and nonesterified fatty acids pre- and postpartum did not differ. High protein induced lower concentration of plasma IL-1 at calving and lowered blood lymphocytes 21 d postpartum, suggestive of a reduced inflammatory status compared with LP. The concentrations of IL-10, tumor necrosis factor alpha, and other hemogram variables did not differ among treatments. Addition of rumen-protected methionine to the HP diet did not alter milk yield but increased fat and total solids concentrations. The rumen-protected methionine had no effect on blood metabolites and immunity markers, with the exception of increased pre-partum insulin concentrations. The data indicate that dairy cows around calving respond positively to an increase in the supply of MP and to rumen-protected methionine supplementation of the HP diet by increasing intake and improving immune status.

Concurrent and carryover effects of feeding blends of protein and amino acids in high-protein diets with different concentrations of forage fiber to fresh cows. 2. Protein balance and body composition

Increasing the supply of metabolizable protein (MP) and improving its AA profile may attenuate body protein mobilization in fresh cows and lead to increased milk production. Increasing the concentration of rumen-undegradable protein (RUP) to increase MP supply and replacing RUP sources from forages instead of nonforage fiber sources may further decrease tissue mobilization if it improves dry matter intake (DMI). Our objective was to determine whether increasing MP concentrations and improving the AA profile at the expense of either nonforage or forage fiber (fNDF) would affect MP balance and empty body (EB) composition (measured using the urea dilution method) in early postpartum dairy cows of different parities. In a randomized block design, 40 primigravid [77 ± 1.5 kg of EB crude protein (CP) at 8 ± 0.6 d before calving] and 40 multigravid (92 ± 1.6 kg of EB CP at 5 ± 0.6 d before calving) Holsteins were blocked by calving date and fed a common prepartum diet (11.5% CP). After calving to 25 d in milk (DIM), cows were fed 1 of 4 diets: (1) a diet deficient in MP meeting 87% of MP requirements (DMP, 17% CP, 24% fNDF), (2) 104% of MP requirements using primarily soy protein to make MP adequate (AMP, 20% CP, 24% fNDF), (3) 110% of MP requirements using a blend of proteins and rumen-protected (RP) AA to make MP adequate (Blend, 20% CP, 24% fNDF), or (4) a diet similar to Blend but substituting added RUP for fNDF rather than nonforage NDF (Blend-fNDF, 20% CP, 19% fNDF). Blend was formulated to have a RUP supply with a similar AA profile to that of casein. Cows were fed a common diet (16.3% CP) from 26 to 50 DIM. Calculated MP balance (supply - requirements) was less than zero for DMP and Blend-fNDF from 1 to 4 wk of lactation (WOL), whereas that for AMP was positive from 1 to 4 WOL and that for Blend was close to zero from 3 to 4 WOL. Daily MP balance was greater from 5 to 7 WOL for DMP compared with AMP and Blend (100 vs. 22 g/d). From -7 to 7 d relative to calving, losses of EB CP were greater for DMP than for AMP and Blend (-121 vs. average of 11 g/d). From 7 to 25 DIM, cows fed AMP (-139 g/d) and Blend-fNDF (-147 g/d) lost EB CP but cows fed Blend (-8 g/d) maintained EB CP. Increased DMI for Blend versus AMP led to reduced losses of EB lipid in primiparous cows from 7 to 25 d relative to calving (-1.0 vs. -1.3 kg/d of EB lipid), whereas lipid mobilization was similar in multiparous cows (average -1.1 kg of EB lipid/d). By 50 DIM, EB lipid and CP were similar across treatments and parities (average 60.2 kg of EB lipid and 81.6 kg of EB CP). Overall, feeding fresh cows a high MP diet with a balanced AA profile improved DMI and attenuated EB CP mobilization, which could partly explain positive carryover effects on milk production for multiparous cows and reduced lipid mobilization for primiparous cows.

Targeted assessment of the metabolome in skeletal muscle and in serum of dairy cows supplemented with conjugated linoleic acid during early lactation

In the dairy cow, late gestation and early lactation are characterized by a complexity of metabolic processes required for the homeorhetic adaptation to the needs of fetal growth and milk production. Skeletal muscle plays an important role in this adaptation. The objective of this study was to characterize the metabolome in skeletal muscle (semitendinosus muscle) and in serum of dairy cows in the context of the physiological changes occurring in early lactation and to test the effects of dietary supplementation (from d 1 in milk onwards) with conjugated linoleic acids (sCLA; 100 g/d; supplying 7.6 g of cis-9,trans-11 CLA and 7.6 g of trans-10,cis-12 CLA per cow/d; n = 11) compared with control fat-supplemented cows (CTR; n = 10). The metabolome was characterized in skeletal muscle samples collected on d 21 and 70 after calving in conjunction with their serum counterpart using a targeted metabolomics approach (AbsoluteIDQ p180 kit; Biocrates Life Sciences AG, Innsbruck, Austria). Thereby 188 metabolites from 6 different compound classes (acylcarnitines, amino acids, biogenic amines, glycerophospholipids, sphingolipids, and hexoses) were quantified in both sample types. In both groups, dry matter intake increased after calving. It was lower in sCLA than in CTR on d 21, which resulted in reduced calculated net energy and metabolizable protein balances. On d 21, the concentrations of dopamine, Ala, and hexoses in the skeletal muscle were higher in sCLA than in CTR. On d 21, the changed metabolites in serum were mainly long-chain (>C24) diacyl phosphatidylcholine PC (PC-aa) and acyl-alkyl phosphatidylcholine (PC-ae), along with lysophosphatidylcholine acyl (lysoPC-a) C26:1 that were all lower in sCLA than in CTR. Supplementation with CLA affected the muscle concentrations of 22 metabolites on d 70 including 10 long-chain (>C22) sphingomyelin (SM), hydroxysphingomyelin [SM(OH)], PC-aa, and PC-ae along with 9 long-chain (>C16) lysoPC-a and 3 metabolites related to amino acids (spermine, citrulline, and Asp). On d 70, the concentrations of lysoPC-a C18:2 and C26:0 in serum were higher in the sCLA cows than in the CTR cows. Regardless of treatment, the concentrations of Ile, Leu, Phe, Lys, His, Met, Trp, and hydroxybutyrylcarnitine (C4-OH) decreased, whereas those of ornithine, Gln, and trans-4-hydroxyproline (t4-OH-Pro) increased from d 21 to 70 in muscle. The significantly changed metabolites in serum with time of lactation were 28 long-chain (>C30) PC-ae and PC-aa, 7 long-chain (>C16) SM and SM(OH), along with lysoPC-a C20:3 that were all increased. In conclusion, in addition to other significantly changed metabolites, CLA supplementation mainly led to changes in muscle and serum concentrations of glycerophospholipids and sphingolipids that might reflect the phospholipid compositional changes in muscle. The metabolome changes observed in sCLA on d 21 seem to be, at least in part, due to the lower DMI in these cows. The changes in the muscle concentrations of AA from d 21 to 70, which coincided with the steady energy and MP balances, might reflect a shift of protein synthesis/degradation balance toward synthesis.

Proteasome activity and expression of mammalian target of rapamycin signaling factors in skeletal muscle of dairy cows supplemented with conjugated linoleic acids during early lactation

The mammalian target of rapamycin (mTOR) is a major regulator of protein synthesis via its main downstream effectors, ribosomal protein S6 kinase (S6K1) and eukaryotic initiation factor 4E binding protein (4EBP1). The ubiquitin-proteasome system (UPS) is the main proteolytic pathway in muscle, and the muscle-specific ligases tripartite motif containing 63 (TRIM63; also called muscle-specific ring-finger protein 1, MuRF-1) and F-box only protein 32 (FBXO32; also called atrogin-1) are important components of the UPS. We investigated 20S proteasome activity and mRNA expression of key components of mTOR signaling and UPS in skeletal muscle of dairy cows during late gestation and early lactation and tested the effects of dietary supplementation (from d 1 in milk) with conjugated linoleic acids (sCLA; 100 g/d; n = 11) compared with control fat-supplemented cows (CTR; n = 10). Blood and muscle tissue (semitendinosus) samples were collected on d -21, 1, 21, and 70 relative to parturition. Dry matter intake increased with time of lactation in both groups. It was lower in sCLA than in CTR on d 21, which resulted in a reduced calculated metabolizable protein balance. Most serum and muscle concentrations of AA followed time-related changes but were unaffected by CLA supplementation. In both groups, serum and muscle 3-methylhistidine (3-MH) concentrations and the ratio of 3-MH:creatinine increased from d -21 to d 1, followed by a decline on d 21. The mRNA abundance of MTOR on d 21 and 70 was greater in sCLA than in CTR. The abundance of 4EBP1 mRNA did not differ between groups but was upregulated in both on d 1. The mRNA abundance of S6K1 on d 70 was greater in CTR than in sCLA, but remained unchanged over time in both groups. The mRNA abundance of FBXO32 (encoding atrogin-1) on d 21 was greater in sCLA than in CTR. The mRNA abundance of TRIM63 (also known as MuRF1) showed a similar pattern as FBXO32 in both groups: an increase from d -21 to d 1, followed by a decline. The mRNA for the α (BCKDHA) and β (BCKDHB) polypeptide of branched-chain α-keto acid dehydrogenase was elevated in sCLA and CTR cows on d 21, respectively, suggesting a role of CLA in determining the metabolic fate of branched-chain AA. For the mTOR protein, no group differences were observed. The abundance of S6K1 protein was greater across all time points in sCLA versus CTR. The antepartum 20S proteasome activity in muscle was elevated in both groups compared with postpartum, probably reflecting the start of protein mobilization before parturition. Plasma insulin concentrations decreased in both groups postpartum but to a greater extent in CTR than in sCLA, resulting in greater insulin concentrations in sCLA than in CTR. Thus, the greater abundance of MTOR mRNA and S6K1 protein in sCLA compared with CTR might be mediated by the greater plasma insulin postpartum. The upregulation of MTOR mRNA in sCLA cows on d 21, despite greater FBXO32 mRNA abundance, may reflect a simultaneous activation of both anabolic and catabolic signaling pathways, likely resulting in greater protein turnover.

The plasma free amino acid dose-response technique: A proposed methodology for determining lysine relative bioavailability of rumen-protected lysine supplements

Estimates of Lys bioavailability of rumen-protected Lys (RP-Lys) supplements are often obtained using in vitro or 2-step in situ techniques, with little to no data determining efficacy and bioavailability in vivo. The objective of this study was to further evaluate and refine the use of the plasma free AA dose-response technique as a method for determining Lys relative bioavailability of RP-Lys supplements. Thirteen dose-response Latin square studies using 87 lactating, ruminally cannulated multiparous Holstein cows (days in milk from 55 to 315 and milk yield from 12 to 62 kg/d at the start of the studies) were conducted to measure the relative bioavailability of RP-Lys supplements. Intestinal (1 study) and abomasal (12 studies) infusions of Lys ranged from 0 to 84 g/d, and experimental periods ranged from 4 to 21 d. Basal diets were formulated to be adequate in metabolizable Met, but varied in predicted metabolizable Lys (5.04 to 6.81% of metabolizable protein). One to 4 daily blood samples were taken from the coccygeal vessels for 1 to 3 consecutive days in each period. Plasma Lys concentration in cows assigned to the control treatment (0 g/d Lys) ranged from 1.83 to 5.21% of total plasma AA, whereas that from cows duodenally or abomasally infused with Lys ranged from 2.53 to 7.51% of total plasma AA. Results from studies involving more than 2 amounts of infused Lys confirmed linearity of response. The following variables were regressed against the plasma Lys dose-response slopes generated from the Lys infusion treatments to examine their effects on the magnitude of the slopes: plasma Lys concentration of the control diet, plasma Lys concentration at the greatest amount of infused Lys, net energy of lactation and metabolizable protein balances, metabolizable protein supply, days in milk, milk yield, milk concentrations of fat, true protein, and lactose, milk true protein yield, and dry matter intake. The variable having the greatest effect on the magnitude of the dose-response slope was the plasma Lys concentration at the greatest amount infused. The relative bioavailability of evaluated RP-Lys supplements using the plasma free AA dose-response technique ranged from 5 to 87%. It was concluded that plasma free Lys increases in a linear fashion to increasing amounts of absorbed Lys and that the dose-response technique is an appropriate technique for evaluating RP-Lys supplements.

Relationships between postruminal casein infusion and milk production, and concentrations of plasma amino acids and blood urea in dairy cows: A multilevel mixed-effects meta-analysis

The relationships between postruminal casein infusion and production variables and concentrations of plasma AA and blood urea were evaluated using multilevel mixed-effects models derived from literature data collected in dairy cows. The data set contained 147 treatment means [i.e., 66 controls (CTL) and 81 casein-infused (CAS) means]. Each CAS mean was paired with its corresponding CTL mean to create 81 mean differences (CAS minus CTL), which were analyzed as absolute and percentage-based units (i.e., percentage increase or decrease in CAS relative to CTL). The primary variable of interest was the difference in estimated metabolizable protein (MP) supply (ΔMP) between CAS and CTL. The other explanatory variables were based on levels in CTL: MP supply, MP balance, the ratio of duodenal microbial protein (MCP) to MP supply (MCPMP), the stage of lactation (early or mid/late) and the type of forage (grass/legume- or corn silage-based). The MP supply and MP balance influenced negatively the relationship between ΔMP and the response of true protein yield. Responses of milk urea, blood urea, and plasma urea cycle AA concentrations were associated positively with ΔMP, indicating that a large amount of infused AA was catabolized to urea. Responses of plasma essential AA concentrations were related positively to ΔMP. The relative effect of ΔMP was highest for responses of plasma His concentration in cows fed grass/legume-based diets and at high MCPMP ratios. This relationship suggests that positive responses of plasma His concentrations are associated with diets relying heavily on microbial protein synthesis (high MCP), low in crude protein (low estimated MP supply), or both. The relationship between ΔMP and responses of plasma group 2 AA (Ile, Leu, Lys, and Val) concentrations was approximately 2 times greater than that for group 1 AA (His, Met, and Phe+Tyr) at mean MCPMP and MP supply. This could reflect the low hepatic removal group 2 AA compared with group 1 AA in dairy cows. Collectively, these results provide novel information on how dietary and cow conditions may alter responses to protein supplementation.

Precalving and early lactation factors that predict milk casein and fertility in the transition dairy cow

Multiparous Holstein cows (n=82) of either high or low genetic merit (GM) (for milk fat + protein yield) were allocated to 1 of 2 diets in a 2×2 factorial design. Diets differed in the ratio of rumen-undegradable protein (RUP) to rumen-degradable protein (37% RUP vs. 15% RUP) and were fed from 21 d precalving to 150 days in milk. This study evaluated the effects of these diets and GM on concentrations of milk casein (CN) variants and aimed to identify precalving and early lactation variables that predict milk CN and protein yield and composition and fertility of dairy cows. It explored the hypothesis that low milk protein content is associated with lower fertility and extended this hypothesis to also evaluate the association of CN contents with fertility. Yields (kg/d) for CN variants were 0.49 and 0.45 of α-CN, 0.38 and 0.34 of β-CN, 0.07 and 0.06 for κ-CN, and 0.10 and 0.09 of γ-CN for high- and low-RUP diets, respectively. Increased RUP increased milk, CN, and milk protein yields. Increased GM increased milk protein and γ-CN yields and tended to increase milk CN yield. The effects of indicator variables on CN variant yields and concentrations were largely consistent, with higher body weight and α-amino nitrogen resulting in higher yields, but lower concentrations. An increase in cholesterol was associated with decreased CN variant concentrations, and disease lowered CN variant yield. A diet high in RUP increased proportion of first services that resulted in pregnancy from 41 to 58%. Increased precalving metabolizable protein (MP) balance decreased the proportion of first services that resulted in pregnancy when evaluated in a model containing CN percentage, milk protein yield, diet, and GM. This finding suggests that the positive effects of increasing dietary RUP on fertility may be curvilinear because cows with a very positive MP balance before calving were less fertile than those with a lower, but positive, MP balance. Prepartum MP balance was important to production and reproductive outcomes, but surprisingly, metabolizable energy balance was not. The hazard of pregnancy in the first 150 d of lactation was 28% lower in cows producing milk with the lowest quartile of protein percentage compared with cows with milk in the upper 3 quartiles. Milk CN percentage was positively associated with improved pregnancy at first service. This study demonstrates the importance of protein metabolism to reproductive performance of the dairy cow.

Effects of rumen-protected methionine, lysine, and histidine on lactation performance of dairy cows

The objective of this study was to evaluate the effects of supplementing a metabolizable protein (MP)-deficient diet with rumen-protected (RP) Met, Lys, and His, individually or combined, on the performance of lactating dairy cows. The experiment was a 9-wk randomized complete block design with 72 Holstein cows. Following a 2-wk covariate period, cows were blocked by days in milk, milk yield, and parity, and randomly assigned to 1 of the following 6 treatments: (1) MP-adequate diet [MPA; +243g/d MP balance, according to the National Research Council (2001) requirements]; (2) MP-deficient diet (MPD; −54g/d MP balance); (3) MPD supplemented with RPMet (MPDM); (4) MPD supplemented with RPLys (MPDL); (5) MPD supplemented with RPHis (MPDH); and (6) MPD supplemented with RPMet, RPLys, and RPHis (MPDMLH). Dry matter intake (DMI), yields of milk and milk components (fat, protein, lactose) and energy-corrected milk (ECM), feed and ECM feed efficiencies, and milk and plasma urea N were decreased by MPD, compared with MPA. Supplementation of the MPD diet with RPLys increased milk protein content and plasma glucose concentration and tended to increase milk urea N. Addition of RPHis tended to increase DMI, increased milk protein concentration, and numerically increased yields of milk fat, protein, and ECM. In addition to the trends for increased DMI and milk fat content, and higher milk protein concentration, supplementation of the 3 RP AA also increased yields of milk fat, protein, and ECM and ECM feed efficiency. Relative to MPA, milk N efficiency tended to be increased by MPD. Concentrations of plasma essential AA (except Met and Thr) were decreased by MPD compared with MPA. Supplementation of RPMet, RPLys, and RPHis increased plasma Met (except for MPDM), Lys, and His concentrations, respectively. Cows fed MPD had lower blood hemoglobin concentration and numerically higher plasma ghrelin than cows fed MPA. Concentration of total saturated fatty acids in milk fat were or tended to be higher for MPD compared with MPA and MPDMLH, respectively. Concentration of total polyunsaturated and yield of milk odd- and branched-chain fatty acids were or tended to be decreased by MPD compared with MPA. Overall, the results of this study confirm our previous data and suggest that His stimulates DMI and the combination of the 3 RP AA (Met, Lys, and His) has the potential to improve milk and milk component yields in dairy cows fed MP-deficient diets.

Casein infusion rate influences feed intake differently depending on metabolizable protein balance in dairy cows: A multilevel meta-analysis

The effects of casein infusion have been investigated extensively in ruminant species. Its effect on responses in dry matter intake (DMI) has been reviewed and indicated no significant effect. The literature reviewed in the current meta-analysis is more extensive and limited to dairy cows fed ad libitum. A total of 51 studies were included in the meta-analysis and data were fitted to a multilevel model adjusting for the correlated nature of some studies. The effect size was the mean difference calculated by subtracting the means for the control from the casein-infused group. Overall, casein infusion [average of 333g of dry matter (DM)/d; range: 91 to 1,092g of DM/d] tended to increase responses in DMI by 0.18kg/d (n=48 studies; 3 outliers). However, an interaction was observed between the casein infusion rate (IR) and the initial metabolizable protein (MP) balance [i.e., supply minus requirements (NRC, 2001)]. When control cows were in negative MP balance (n=27 studies), responses in DMI averaged 0.28kg/d at mean MP balance (−264g/d) and casein IR (336g/d), and a 100g/d increment in the casein IR from its mean increased further responses by 0.14kg/d (MP balance being constant), compared with cows not infused with casein. In contrast, when control cows were in positive MP balance (n=22 studies; 2 outliers), responses in DMI averaged −0.20kg/d at mean casein IR (339g/d), and a 100g/d increment in the casein IR from its mean further decreased responses by 0.33kg/d, compared with cows not infused with casein. Responses in milk true protein yield at mean casein IR were greater (109 vs. 65g/d) for cows in negative vs. positive MP balance, respectively, and the influence of the casein IR on responses was significant only for cows in negative MP balance. A 100g/d increment in the casein IR from its mean increased further responses in milk true protein yield by 25g/d, compared with cows not infused with casein. Responses in blood urea concentration increased in casein studies (+0.59mM) and the influence of the casein IR was greatest for cows in positive MP balance (0.26 vs. 0.11mM per 100g/d increment). Responses in DMI were also correlated negatively with responses in blood urea concentration only for cows in positive MP balance. Together, these results suggest an association between satiety and deamination and oxidation of AA supplied in excess of requirements for cows in positive MP balance. Therefore, casein stimulated appetite in cows fed MP-deficient diets possibly via the supply of orexigenic AA or through a pull effect in response to an increased metabolic demand. Conversely, casein induced satiety in cows fed diets supplying MP in excess of requirements. Not precluding other factors involved in satiety (e.g., insulin, gut peptides), casein could have increased the supply of AA (e.g., Ser, Thr, Tyr), which might depress appetite at the brain level or increase the deamination and the oxidation of AA in oversupply in agreement with the hepatic oxidation theory.

Metabolic and endocrine profiles and hepatic gene expression of Holstein cows fed total mixed ration or pasture with different grazing strategies during early lactation.

BackgroundIn dairy mixed production systems, maximizing pasture intake and total mixed ration (TMR) supplementation are management tools used to increase dry matter and energy intake in early lactation. The objective was to evaluate metabolic and endocrine profiles and hepatic gene expression of Holstein cows fed either TMR ad libitum (without grazing) or diets combining TMR (50 % ad libitum DM intake) and pasture with different grazing strategies (6 h in one grazing session or 9 h in two grazing sessions) in early lactation. Pluriparous cows were grouped by calving date, blocked within group by body weight and body condition score (BCS) and randomly assigned to one of three feeding strategies from calving (day 0) to 60 days postpartum: control cows fed TMR ad libitum (G0; confined cows fed 100 % TMR without access to pasture), pasture grazing with 6 h of access in one session supplemented with 50 % TMR (G1), and 9 h of access in two sessions supplemented with 50 % TMR (G2).ResultsNet energy (NE), but not metabolizable protein (MP), demands for maintenance and/or milk increased in G2 when compared with G1 and G0 cows, respectively. However, NE and MP balances were lower in G1 and G2 than G0 cows. Cow BCS at +55 days was greater in G0 than G2 cows and probability of cows cycling during the first month was greater in G0 and G1 than G2 cows. During the postpartum period, non-esterified fatty acids were greater in G1 than G2 and G0 and β-hydroxybutyrate was greater in G1 and G2 than G0 cows. Plasma insulin was greater and insulin-like growth factor (IGF)-I tended to be greater in G0 than G2 cows, leptin was greater in G2 and G0 and adiponectin were greater in G2 cows. Hepatic expression of growth hormonereceptor-1A and IGF1 mRNA decreased while IGF binding proteins 1,2,4,5 and 6 (IGFBP) mRNA as well as mRNA expression of insulin, leptin (LEPRb) and adiponectin-2 receptors increased from pre to postpartum in all cows. However, only hepatic IGFBP6 and LEPRb mRNA were greater in G2 than G0 and G1 cows, respectively.ConclusionMetabolic-endocrine profiles of cows with different feeding strategies in early lactation reflected not only changes in milk energy output and energy balance but also in walking and grazing activity. Concentrations of insulin and IGF-I were increased in G0 cows whereas plasma adiponectin and both, insulin and leptin sensitivity were improved G2 cows. Increased NE demands in G2 cows when compared to G1 and G0 cows, implied a metabolic stress that impacted negatively on reproductive function.Electronic supplementary materialThe online version of this article (doi:10.1186/s13028-015-0163-6) contains supplementary material, which is available to authorized users.

Effect of dietary protein level and rumen-protected amino acid supplementation on amino acid utilization for milk protein in lactating dairy cows

This study investigated the effect of metabolizable protein (MP) supply and rumen-protected (RP) Lys and Met supplementation on productivity, nutrient digestibility, urinary N losses, apparent total-tract digestibility of dietary AA, and the efficiency of AA utilization for milk protein synthesis in dairy cows. The experiment was conducted with 8 ruminally cannulated Holstein cows in a replicated 4×4Latin square design trial with 21-d periods. Treatments were (1) MP-adequate diet (AMP; MP balance of −24g/d); (2) MP-deficient diet (DMP; MP balance of −281g/d); (3) DMP supplemented with 100g of RPLys/cow per day (estimated digestible Lys supply=24g/d; DMPL; MP balance of −305g/d); and (4) DMPL supplemented with 24g of RPMet/cow per day (estimated digestible Met supply=15g/d; DMPLM; MP balance of −256g/d). Diet had no effect on total-tract nutrient digestibility, milk production, and milk composition, but the DMP diets decreased urinary N excretion and the ammonia emitting potential of manure. Plasma Met concentration was increased by DMPLM compared with AMP. Supplementation with RPLys had no effect on plasma Lys. Concentration of most AA in milk protein was increased or tended to be increased by DMPLM compared with DMPL. Except for the AA supplemented as RPAA (i.e., Met and Lys), apparent total-tract digestibility of all dietary AA was generally greater for the DMP diets and ranged from 33% (Arg, AMP diet) to 67% (Thr, DMPL diet). Apparent recovery of dietary AA in milk protein followed the same trends, being greater for the DMP diets than AMP and generally lower for Lys and Met with the RPAA-supplemented diets versus AMP and DMP. The RPAA were apparently not used for milk protein synthesis in the conditions of this experiment. The AA recoveries in milk protein varied from around 17% (Ala) to 70% (Pro). Milk protein recoveries of essential AA (EAA) were around 54% for the DMP diet and 49% for AMP. The estimated efficiency of utilization of digestible EAA for milk protein synthesis was generally greater for the DMP diets compared with AMP. In this trial, blood plasma Lys and Met were labeled by abomasal pulse-dose of 15N-Lys and 13C-Met (respectively). Analysis of the 15N-Lys and 13C-Met decay curves in plasma indicated trends for a faster extraction of Lys and Met from plasma for the MP-deficient diets, compared with AMP. Overall, this study confirmed conclusions from previous analyses that the efficiency of utilization of dietary EAA will increase with decreasing MP-AA supply.

Abomasal protein infusion in postpartum transition dairy cows: Effect on performance and mammary metabolism

The effect of increasing the postpartum metabolizable protein (MP) supply on performance and mammary metabolism was studied using 8 Holstein cows in a complete randomized design. At parturition, cows were assigned to abomasal infusion of water (CTRL) or casein (CAS). Arterial and epigastric venous blood samples were taken 14 d before expected parturition and at 4, 15, and 29 d in milk (DIM). To compensate previously estimated deficiency of essential AA and to avoid oversupply, casein protein infusion was graduated with 696±1, 490±9, and 212±10g/d at 4, 15 and 29 DIM, respectively. Dry matter intake was unaffected by CAS. Compared with CTRL, MP supply was greater at 4 DIM with CAS but did not differ by 29 DIM. Milk yield was greater with CAS (+7.2±1.3kg/d from 1 to 29 DIM). Milk protein yield was greater with CAS at 4 DIM and averaged 1,664±39g/d compared with 1,212±86g/d for CTRL, but did not differ at 29 DIM (1,383±48g/d). The ratio of MP total supply to requirement was numerically greater at 4 DIM for CAS compared with CTRL, indicating less postpartum protein deficiency. In contrast, a greater net energy deficiency tended to be induced with CAS, but the greater milk yield allowed a large part of mobilized fat to be secreted in milk. Arterial concentration of total essential AA increased sharply after parturition for CAS compared with slight decreases for CTRL. The patterns of arterial concentrations combined with arterial-mammary venous concentration differences indicated that Lys, Leu, and Tyr were the first-limiting AA at 4 DIM with CTRL. Mammary plasma flow was unaffected by treatment, indicating similar perfusion of mammary tissue. The greater milk yield with CAS was associated with greater mammary uptake of individual essential AA, tendencies to greater uptake of glucose, lactate, and β-hydroxybutyrate, whereas uptakes of volatile fatty acids were unaffected. Despite similar MP supply by 29 DIM, milk and lactose yields were greater with CAS indicating a persistent response to increased postpartum MP supply. In conclusion, the postpartum MP deficiency can have a substantial negative effect in dairy cows as the major outcome of increasing the postpartum MP supply was increased milk, milk protein, and lactose yield, as well as an enhanced MP balance. Potential positive effects for other body functions than milk synthesis are discussed. Future investigations are needed to delineate how to transfer the effect into practical feeding strategies.

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 (R2=0.99).

Rumen-protected lysine, methionine, and histidine increase milk protein yield in dairy cows fed a metabolizable protein-deficient diet

The objective of this experiment was to evaluate the effect of supplementing a metabolizable protein (MP)-deficient diet with rumen-protected (RP) Lys, Met, and specifically His on dairy cow performance. The experiment was conducted for 12wk with 48Holstein cows. Following a 2-wk covariate period, cows were blocked by DIM and milk yield and randomly assigned to 1 of 4 diets, based on corn silage and alfalfa haylage: control, MP-adequate diet (ADMP; MP balance: +9 g/d); MP-deficient diet (DMP; MP balance: −317g/d); DMP supplemented with RPLys (AminoShure-L, Balchem Corp., New Hampton, NY) and RPMet (Mepron; Evonik Industries AG, Hanau, Germany; DMPLM); and DMPLM supplemented with an experimental RPHis preparation (DMPLMH). The analyzed crude protein content of the ADMP and DMP diets was 15.7 and 13.5 to 13.6%, respectively. The apparent total-tract digestibility of all measured nutrients, plasma urea-N, and urinary N excretion were decreased by the DMP diets compared with ADMP. Milk N secretion as a proportion of N intake was greater for the DMP diets compared with ADMP. Compared with ADMP, dry matter intake (DMI) tended to be lower for DMP, but was similar for DMPLM and DMPLMH (24.5, 23.0, 23.7, and 24.3kg/d, respectively). Milk yield was decreased by DMP (35.2kg/d), but was similar to ADMP (38.8kg/d) for DMPLM and DMPLMH (36.9 and 38.5kg/d, respectively), paralleling the trend in DMI. The National Research Council 2001model underpredicted milk yield of the DMP cows by an average (±SE) of 10.3±0.75kg/d. Milk fat and true protein content did not differ among treatments, but milk protein yield was increased by DMPLM and DMPLMH compared with DMP and was not different from ADMP. Plasma essential amino acids (AA), Lys, and His were lower for DMP compared with ADMP. Supplementation of the DMP diets with RP AA increased plasma Lys, Met, and His. In conclusion, MP deficiency, approximately 15% below the National Research Council requirements from 2001, decreased DMI and milk yield in dairy cows. Supplementation of the MP-deficient diet with RPLys and RPMet diminished the difference in DMI and milk yield compared with ADMP and additional supplementation with RPHis eliminated it. As total-tract fiber digestibility was decreased with the DMP diets, but DMI tended to increase with RP AA supplementation, we propose that, similar to monogastric species, AA play a role in DMI regulation in dairy cows. Our data implicate His as a limiting AA in high-producing dairy cows fed corn silage- and alfalfa haylage-based diets, deficient in MP. The MP-deficient diets clearly increased milk N efficiency and decreased dramatically urinary N losses.

Effects of dietary protein concentration and coconut oil supplementation on nitrogen utilization and production in dairy cows

The objective of this study was to investigate the effect of metabolizable protein (MP) deficiency and coconut oil supplementation on N utilization and production in lactating dairy cows. The hypothesis of the study was that a decrease in ruminal protozoal counts with coconut oil would increase microbial protein synthesis in the rumen, thus compensating for potential MP deficiency. The experiment was conducted for 10 wk with 36 cows (13 primiparous and 23 multiparous), including 6 ruminally cannulated cows. The experimental period, 6 wk, was preceded by 2-wk adaptation and 2-wk covariate periods. Cows were blocked by parity, days in milk, milk yield, and rumen cannulation and randomly assigned to one of the following diets: a diet with a positive MP balance (+44g/d) and 16.7% dietary crude protein (CP) concentration (AMP); a diet deficient in MP (−156g/d) and 14.8% CP concentration (DMP); or DMP supplemented with approximately 500g of coconut oil/head per day (DMPCO). Ruminal ammonia tended to be greater and plasma urea N (20.1, 12.8, and 13.1mg/dL, for AMP, DMP, and DMPCO diets, respectively) and milk urea N (12.5, 8.3, and 9.5mg/dL, respectively) were greater for AMP compared with DMP and DMPCO. The DMPCO diet decreased total protozoa counts (by 60%) compared with DMP, but had no effect on the methanogens profile in the rumen. Total tract apparent digestibility of dry matter and CP was decreased by DMP compared with AMP. Fiber digestibility was lower for both DMP and DMPCO compared with AMP. Urinary N excretion was decreased (by 37%) by both DMP and DMPCO compared with AMP. The DMP and DMPCO diets resulted in greater milk N efficiency compared with AMP (32.0 and 35.1 vs. 27.6%, respectively). Milk yield was decreased by both DMP and DMPCO compared with AMP (36.2, 34.4, and 39.3kg/d, respectively) and coconut oil supplementation suppressed feed intake and caused milk fat depression. Coconut oil supplementation decreased short-chain fatty acid (C4:0, C6:0, and C8:0) concentration and increased medium-chain (C12:0 and C14:0) and total trans fatty acids in milk. Overall, the MP-deficient diets decreased N losses, but could not sustain milk production in this study. Coconut oil decreased feed intake and similar to DMP, suppressed fiber digestibility. Despite decreased protozoal counts, coconut oil had no effect on the methanogen population in the rumen.

Effect of degradable intake protein supplementation in finishing diets containing dried distillers grains or wet distillers grains plus solubles on performance and carcass characteristics

<h2>ABSTRACT</h2> Two finishing trials were conducted to determine whether supplemental degradable intake protein is necessary to optimize performance in dry-rolled corn (DRC) based finishing diets containing distillers grains. Diets without urea were deficient in degradable intake protein based on the NRC (1996) recommendations, yet all diets had positive metabolizable protein balances. In Exp. 1, individually fed, crossbred heifers (n = 58; BW=383 ± 25 kg) were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments. Dietary treatments contained 10 or 20% dried distillers grains replacing DRC, with or without 0.80 or 0.63% urea, respectively (DM basis). Blood samples were analyzed for BUN. Heifers receiving urea or 20% dried distillers grains (P < 0.01) had greater BUN concentration than did heifers not receiving urea or fed 10% dried distillers grains, respectively. In Exp. 2, pen-fed, crossbred steers (n = 336; initial BW=365 ± 31 kg) were assigned randomly within block to 1 of 24 pens in a randomized complete block design with 3 treatments. Dietary treatments were 0, 0.5, or 1.0% urea added to a DRC diet containing 25% wet distillers grains plus solubles (DM basis). Marbling tended to decrease linearly (P = 0.09) as urea level increased. However, no other significant differences (P > 0.10) were observed for performance or carcass characteristics for either experiment. These data indicate supply of ruminal degradable intake protein was adequate to maintain performance when at least 10% distillers grains was included in DRC diets.