Comparison of the in situ and in vivo intestinal disappearance of ruminally protected methionine

We designed a switchback expriment to compare the intestinal disappearance of ruminally protected Met determined in situ or in vivo over three consecutive periods. Four nonlactating Holstein heifers (477+/-36 kg) with cannulae in the rumen, duodenum, and ileum were fed a diet based on timothy silage to meet requirements for maintenance (dry matter intake = 8.72+/-0.15 kg). A total of 16 bags, containing 1.5 g of ruminally protected Met, were incubated in the rumen (4.5 h) of each cow and transferred to an acid-pepsin solution to simulate the abomasum (2.5 h). Following each incubation, bags were recovered and three bags were dried at 55 degrees C and analyzed for Met. Remaining bags were introduced directly into the duodenal or ileal cannula for the in situ method, while for the in vivo method, the content of remaining bags was transferred into gelatin capsules before their introduction in the duodenal or ileal cannula. Spot samples of digesta were collected during a 96-h period, with Co-EDTA and Cr-mordanted fiber used as indigestible markers to estimate in vivo digestibility. The disappearance of Met in the small intestine determined in vivo tended to be higher than in situ (74.45 vs. 43.65+/-1.79%). Our results indicate that when used to assess intestinal availability of ruminally protected Met, the mobile nylon bag technique can underestimate the true bioavailability of Met.     

Ruminal degradability, intestinal disappearance, and plasma methionine response of rumen-protected methionine in dairy cows.

Bioavailability of Met from a rumen-protected Met product was evaluated in two experiments using three ruminally and duodenally cannulated lactating (experiment 1) and nonlactating (experiment 2) dairy cows. In the first experiment, the ruminal in situ and mobile bag technique was used to assess ruminal degradability and intestinal disappearance of Met from the protected Met product. Effective ruminal degradability of Met at a ruminal outflow rate of 0.11/h was 21.7%. Combining effective ruminal degradability with intestinal digestibility yielded an estimate of Met availability of 25%. In the second experiment, designed as a 3 x 3 Latin square, Met availability was assessed by determining the response of plasma Met to supplementation of the protected Met product relative to that of duodenally administered Met. The periods were 1 wk with cows fed a meal containing 0, 20, or 63 g of protected Met on d 1 and infused intraduodenally with 10.7 g of Met on d 4. Blood was collected at various times relative to the time of oral dosing and the commencement of the duodenal infusion. Plasma Met response measured as area under the curve increased linearly with increasing protected Met. The response of plasma Met increased by 33 and 65.5% of the control values for 20 and 63 g of protected Met, respectively. Intestinal bioavailability of Met in the protected Met product ranged from 22 to 34%.     

Varying protein and starch in the diet of dairy cows. I. Effects on ruminal fermentation and intestinal supply of nutrients

The main objective of this experiment was to examine the effects of the percentage and source of crude protein (CP) and the amount of starch in the diet of dairy cows on ruminal fermentation, nutrient passage to the small intestine, and nutrient digestibility. For this purpose, 6 multiparous Holstein cows fistulated in the rumen and duodenum that averaged 73 d in milk were used in a 6 × 6 Latin square design with a 2 × 3 factorial arrangement of treatments. Two sources of CP [solvent-extracted soybean meal (SBM) and a mixture of SBM and a blend of animal-marine protein supplements plus ruminally protected Met (AMB)] and 3 levels of dietary protein (about 14, 16, and 18%) were combined into 6 treatments. On a dry matter (DM) basis, diets contained 25% corn silage, 20% alfalfa silage, 10% cottonseed, 26.7 to 37% corn grain, and 4 to 13.5% protein supplement. Intakes and digestibilities in the rumen and total tract of DM, organic matter, acid and neutral detergent fiber were unaffected by treatments. Increasing dietary CP from 14 to 18% decreased the intake and apparent ruminal and total tract digestion of starch, but increased the proportion of starch consumed by the cows that was apparently digested in the small intestine. At 14% CP, starch intake and total tract digestion were higher for the AMB diet than for the SBM diet, but the opposite occurred at 16% CP. Across CP sources, increasing CP in the diet from 14 to 18% increased the intakes of N and amino acids (AA), and ruminal outflows of nonammonia N, nonammonia nonmicrobial N, each individual AA except Met, total essential AA, and total AA. Across CP percentages, replacing a portion of SBM with AMB increased the intake of Met and Val and decreased the concentration of ammonia N in the rumen, but did not affect the intake of other essential AA or the intestinal supply of any essential AA and starch. The ruminal outflow of microbial N, the proportional contribution of Lys and Met to total AA delivered to the duodenum, and milk yield were unaffected by treatments. Data suggest that the intake of N by high-producing dairy cows that consume sufficient energy and other nutrients to meet their requirements can be decreased to about 600 to 650 g daily without compromising the supply of metabolizable protein if the source and amount of dietary CP and carbohydrate are properly matched.     

Mammary uptake, portal-drained visceral flux, and hepatic metabolism of free and peptide-bound amino acids in cows fed steam-flaked or dry-rolled sorghum grain diets

Our objectives were to measure net fluxes of free AA (FAA) and peptide-bound AA (PBAA) across portal-drained viscera, liver, splanchnic tissues, and mammary tissues, and milk AA output of lactating Holstein cows (n = 8, 86 ± 8 d in milk). Cows were fed an alfalfa-based total mixed ration containing 40% steam-flaked (SFS) or dry-rolled (DRS) sorghum grain. The total mixed rations were offered at 12-h intervals in a crossover design. Blood samples were obtained from indwelling catheters in portal, hepatic, and mammary veins and from mesenteric or costoabdominal arteries every 2 h from each cow and diet. Intake of dry matter was 17.9 and 18.6 kg/d of the SFS and DRS diets, respectively, but dropped to 16.3 kg/d for cows fed the SFS diet in the last 3 experimental days, sampling day included. Milk and milk crude protein yields (kg/12-h sampling) were 13.85 vs. 13.25 and 0.425 vs. 0.396 for cows fed SFS or DRS, respectively, and were not affected by the considerable drop in dry matter intake of cows fed the SFS diet during the last 3 experimental days. The portal-drained visceral flux of total essential FAA was 417 and 442 g/12 h (SEM 63) in cows fed SFS and DRS, respectively. However, the portal-drained visceral flux of 7 essential PBAA out of the 9 determined was numerically greater in cows fed the SFS diet, and total essential PBAA in that treatment was 77.4 ± 22.2 compared with 35.4 ± 50.2 g/12 h for cows fed the DRS diet. This phenomenon was again observed in a greater total splanchnic flux (FAA + PBAA) of 462 and 371 g/12 h in SFS- and DRS-fed cows, respectively. Mammary uptake of essential AA from both pools (free and peptide bound), and recovery of essential AA in milk, was again numerically higher in SFS-fed cows. In addition to FAA, quantifying the contribution of PBAA may improve our understanding of tissue use of AA substrates, and this may ultimately lead to improved diet formulations with respect to intestinal absorption and mammary uptake of AA.     

Portal drained visceral flux, hepatic metabolism, and mammary uptake of free and peptide-bound amino acids and milk amino acid output in dairy cows fed diets containing corn grain steam flaked at 360 or steam rolled at 490 g/L

Objectives were to measure net fluxes of free (FAA) and peptide bound amino acids (AA) (PBAA) across portal-drained viscera (PDV), liver, splanchnic, and mammary tissues, and of milk AA output of lactating Holstein cows (n = 6, 109 +/- 9 d in milk) as influenced by flaking density of corn grain. Cows were fed alfalfa-based total mixed ration (TMR) containing 40% steam-flaked (SFC) or steam-rolled corn (SRC) grain. The TMR were offered at 12-h intervals in a crossover design. Six sets of blood samples were obtained from indwelling catheters in portal, hepatic, and mammary veins and mesenteric or costoabdominal arteries every 2 h from each cow and diet. Intake of dry matter (18.4 +/- 0.4 kg/d), N, and net energy for lactation were not altered by corn processing. Milk and milk crude protein yields (kg/12-h sampling) were 14.2 vs. 13.5 and 0.43 vs. 0.39 for cows fed SFC or SRC, respectively. The PDV flux of total essential FAA was greater (571.2 vs. 366.4 g/12 h, SEM 51.4) in cows fed SFC. The PDV flux of total essential PBAA was 69.3 +/- 10.8 and 51.5 +/- 13.2 g/12 h for cows fed SFC and SRC, respectively, and differed from zero, but fluxes of individual PBAA rarely differed between treatments. Liver flux of essential FAA was greater in cows fed SRC, but only the PBAA flux in cows fed SRC differed from zero. Splanchnic flux of FAA and PBAA followed the pattern of PDV flux, but variation was greater. Mammary uptake (g/12 h) of total essential FAA was greater in cows fed SFC than SRC (224.6 vs. 198.3, SEM 7.03). Mammary uptake of essential PBAA was 25.0 vs. 15.1, SEM 5.2, g/12 h for cows fed SFC or SRC, respectively, and differed from zero in half of the PBAA. Milk output of EAA was 187.8 vs 175.4, SEM 4.4 g/12 h in cows fed SFC and SRC, respectively, and output of most essential AA consistently tended to be greater in cows fed SFC. It is apparent that PBAA comprise a portion of total AA flux across PDV and are affected by grain processing. Further, this pool supplies an important component of AA taken up by the mammary gland. Quantifying the contribution of PBAA may improve diet formulation with respect to intestinal absorption and mammary uptake of AA.     

Effect of micronizing full fat canola seed on amino acid disappearance in the gastrointestinal tract of dairy cows

Ruminal and total tract digestion of the amino acids (AA) in full fat canola seed was studied in two in situ experiments with three nonlactating, ruminally and duodenally fistulated dairy cows. Whole, full fat canola seed was hand-cracked or micronized (an infrared heat treatment) for 90 s and then studied in that form or after grinding to pass a 1.25-mm sieve. In the first experiment, the four sample types were ruminally incubated in nylon bags for up to 96 h. In the second experiment, they were sealed in mobile nylon bags, incubated ruminally for 16 h, placed in acidified pepsin for 1 h, and then inserted into duodenal cannulas for passage through the intestine. Amino acids in the canola seed and in the residues from in situ incubations were analyzed by HPLC. Micronization reduced ruminal disappearance of total AA and essential AA from full fat canola seed. Degradation kinetics from Experiment 1 indicated reduced soluble fraction and increased slowly degradable fraction of both total AA and essential AA following micronization. Micronization reduced disappearances of total AA and essential AA from whole canola seed in the total digestive tract but did not affect total tract digestion of total AA or essential AA in ground seed. Intestinal disappearance of total AA and essential AA from both whole and ground full fat canola seed were increased by micronization. Micronizing canola seed may be of value in improving AA utilization in ruminants.     

Effect of distillers dried grains with solubles diet supplemented with rumen‐protected lysine and methionine on milk production and milk amino acids in the lactating cow

This study was to quantify the milk protein and amino acids (AA) in response to rumen‐protected lysine (Lys) and methionine (Met) supplementation in dairy cows fed distillers dried grains with solubles (DDGS) diet. Forty cows were divided into five dietary treatments: soybean meal diet (SM), DDGS diet and DDGS diets with 40 g/day Lys (DL), 20 g/ day Met dry matter (DM) and 20 g/ day Met plus 40 g/ day Lys (DLM), separately. Compared with SM, DL and DM groups, DLM group increased milk yield, 4% fat‐corrected milk, energy‐corrected milk yield and protein in milk. DDGS treatments increased total AA and the essential AA in milk. Results indicated that dietary DDGS increased intestinal supplies of Lys and Met.     

Forage proportion and particle length affects the supply of amino acids in lactating dairy cows

This study was conducted to evaluate the effects of dietary factors that alter ruminal fermentability on intake, duodenal flows and intestinal digestibility of individual amino acids (AA) in lactating dairy cows. The experiment was designed as a 4×4 Latin square using 4 ruminally and duodenally cannulated lactating dairy cows. Treatments were arranged in a 2×2 factorial design; 2 forage particle lengths (FPL) of alfalfa silage (short and long) were combined with low (35:65) and high (60:40) forage-to-concentrate ratio (F:C; dry matter basis). Four diets were formulated using 2 cuts of alfalfa silage [short (7.9 mm) and long (19.1mm)], combined with 2 ratios of forage to barley grain concentrate (35:65 and 60:40). Overall, the interactions between dietary F:C and FPL on intake, duodenal flows, and intestinal digestibility of AA were marginal. Intakes of total AA and nonessential AA were not different between low- and high-F:C diets, whereas that of essential AA tended to be less with high-F:C diet as a result of lower intakes of Met, Phe, Arg, and His. The flows of total AA and microbial AA were reduced by 22 and 19%, respectively, with increasing F:C ratio in the diets due to consistently reduced flows of individual AA, whereas AA profiles (% of AA-N) of the duodenal protein were not different. Altering F:C from 35:65 to 60:40 decreased the intestinal digestibility of Ile, Leu, Thr, Val, Ala, Cys, and Ser, and consequently, tended to decrease the digestibility of total AA, essential AA, and nonessential AA. Intakes of total AA, essential AA, and nonessential AA were overall not affected by dietary FPL so FPL did not affect the flows or intestinal digestibility of AA. These results indicate that increasing dietary F:C ratio decreased overall AA supply because flow to the duodenum and intestinal digestibility of AA were decreased. However, increasing FPL had no effect on AA supply. The measured duodenal flows of AA were consistent with the predictions of the Cornell Net Carbohydrate and Protein System model for the low-forage diet, and were consistent with the National Research Council model for the high-forage diet. Furthermore, the digestibility of individual AA in the intestine varied considerably, regardless of dietary treatment. The results revealed the necessity to consider the both flows and digestibility of individual AA when optimizing ration formulation to meet AA requirements of dairy cows.     

Evaluation of ruminally protected methionine and lysine or blood meal and fish meal as protein sources for lactating Holsteins.

Forty lactating Holstein cows averaging 55 days in milk were used in a randomized block designed experiment to evaluate the effectiveness of ruminally protected Met and Lys compared with that of ruminally undegradable protein for supporting lactation. Cows were fed total mixed diets for 15 wk. Diets were formulated to be isonitrogenous with the same base ingredients resulting in base crude protein percentage of 15.5. Supplemental crude protein supplied by urea, soybean meal, or a 50:50 (wt/wt) mixture of fish and blood meal increased total dietary nitrogen to 18.0% of diet DM. Two additional diets consisted of the basal diets soybean meal and urea, which were supplemented with ruminally protected DL-Met and Lys-HCL at 10 and 25 g/d, respectively (soybean meal + amino acids (AA), urea + AA). Mean measures of dry matter intake, milk yield, milk protein percentage, and milk fat percentage were not affected by protein supplement. Milk protein yield, milk fat yield, casein yield, and casein percentage also were not affected by source of supplemental protein. Results indicate that at the level of crude protein intake relative to milk production in this experiment, the source of protein did not affect lactational performance.     

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

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

Protected methionine supplementation to a barley-based diet for cows during early lactation

Formulation of diets using barley and soybean meal should result in a diet that supplies less methionine than conventional corn and soybean meal diets. To evaluate this further, 28 high producing Holstein cows (10 primiparous and 18 multiparous) were fed a barley soybean meal diet without or with 15 g of added DL-methionine as 50 g of ruminally protected methionine product during wk 4 to 16 postpartum. Cows were fed a 15% CP total mixed diet consisting of (dry matter basis) 50% concentrate mix, 45% corn silage, and 5% chopped alfalfa hay. Yields of milk (30.3 and 29.8 kg/d), 4% fat-corrected milk (26.0 and 25.6 kg/d), and solids-corrected milk (26.3 and 25.9 kg/d) were similar for cows fed diets without or with added methionine. Percent of milk fat (3.08 and 3.16%) and solids-not-fat (8.66 and 8.71%) were similar, but percent protein (2.75 and 2.87%) was higher, from cows fed ruminally protected methionine. Methionine concentrations in arterial and venous serum were not significantly elevated by feeding ruminally protected methionine. Supplemental ruminally protected methionine did not increase milk production but did increase milk protein percentages in cows fed barley-based diets.     

Protected methionine supplementation with extruded blend of soybeans and soybean meal for dairy cows

Methionine may be the first amino acid limiting milk production in early lactation cows. To evaluate this further, 23 high producing Holstein cows (9 multiparous and 14 primiparous) were fed an extruded blend of soybeans and soybean meal (40:60) without or with 15 g of added DL-methionine as 50 g of ruminally protected methionine product during wk 4 to 16 postpartum. Cows were fed a 15.8% crude protein total mixed ration consisting of 30% (dry basis) corn silage, 15% alfalfa hay, and 55% concentrate mix. Covariant-adjusted yields of milk (35.3 and 33.9 kg/d) and solids-corrected milk (29.3 and 28.2 kg/d) were lower for cows fed ruminally protected methionine, whereas yields of 4% fat-corrected milk (28.2 and 27.4 kg/d) were similar. Percentages of fat (2.68 and 2.69) and solids-not-fat (8.82 and 8.83) were similar, and percentages of protein (2.86 and 2.90) were higher from cows fed supplemental methionine. Dry matter intakes (20.5 and 21.6 kg/d) were higher for cows fed ruminally protected methionine. Methionine concentrations in arterial and venous serum were elevated slightly by feeding supplemental methionine. Although methionine was still the first-limiting amino acid as calculated by two different methods, supplementation of this diet with ruminally protected methionine did not increase production of early lactation cows.     

Assessing bioavailability of ruminally protected methionine and lysine prototypes

Met and Lys are essential AA that can limit lactational performance in dairy cattle fed protein-sufficient diets. Thus, there is industry demand for ruminally protected (RP) sources of Met and Lys. One method of providing ruminal protection for Met and Lys is lipid encapsulation. The objective of this work was to assess 3 lipid-encapsulated Met prototypes (P1, P2, and P3) and 1 Lys prototype (P4) to determine ruminal protection, small intestine absorption (experiment 1), and animal production responses (experiment 2). Ruminal protection was estimated from 8-h in situ retention during ruminal incubation and intestinal absorption from plasma appearance after an abomasal bolus of the in situ retentate. Blood samples were collected over time to determine plasma Met and Lys concentration responses compared with unprotected Lys and Met infused abomasally. The prototypes were not exposed to the total diet or subjected to typical feed handling methods before evaluation. The bioavailability of P1, P2, and P3 Met prototypes was found to be 14, 21, and 18% of the initial AA material, respectively. The RP-Lys prototype had a bioavailability of 45%. To evaluate production responses, 20 Holstein cows were randomly assigned to 2 trials (n = 10 each) in a replicated Latin square design with 14-d periods. The base diet was predicted to be deficient in metabolizable Met (?14.8 g/d) and Lys (?16.1 g/d) per the Cornell Net Carbohydrate and Protein System (version 6.55). In the Met trial, the base diet was supplemented with RP-Lys to meet Lys requirements, and treatments were as follows: no added RP-Met (NCM), NCM plus Smartamine M (SM; Adisseo, Alpharetta, GA), and NCM plus P1, P2, or P3 at 148% of the Met content of SM. In the Lys trial, the base diet was supplemented with RP-Met to meet the Met requirement, and treatments were as follows: no added Lys (NCL), NCL plus AjiProL (AL; Ajinomoto Heartland Inc., Chicago, IL), and NCL plus P4 at 55, 78, or 102% of the reported absorbed Lys in AL. All products were top dressed on the diet without prior mixing or extended exposure to the rest of the diet. Milk protein concentration significantly increased when diets were supplemented with P2, P3, or SM (3.12, 3.12, and 3.11%, respectively) compared with NCM (3.02%). Only P1 (3.04%) was significantly lower than SM. Prototype P2 had the greatest numerical milk protein output response among the 3 RP-Met prototypes, suggesting that it may have had the greatest efficacy when supplemented into these rations. There was a numerical milk protein concentration response to AL and a linear increase in milk protein concentration for P4. The P4 and AL treatments resulted in comparable milk protein production regardless of P4 dose.     

Effect of ruminally protected methionine on splanchnic metabolism of amino acids in lactating dairy cows

The effect of ruminally protected Met (RPM) on splanchnic metabolism was measured in 3 primiparous and 3 multiparous Holstein cows. Doses of RPM (0, 36, and 72 g/d) were tested in a replicated 3 × 3 Latin square design, over 3 consecutive 14-d experimental periods. A mixed ration was fed in 12 equal meals per d (average dry matter intake: 17.5 ± 0.08 kg/d). Indwelling catheters were surgically implanted in the mesenteric artery and the portal and hepatic veins for blood collection, as well as in 2 distal branches of the mesenteric vein for infusion of p-aminohippurate to determine blood flow. On d 14 of each period, a temporary catheter was inserted into a mammary vein and 6 hourly blood samples were collected to determine plasma concentrations of metabolites, hormones, and their respective fluxes across the splanchnic bed and mammary glands. Yields of milk (32.8, 32.0, and 32.9 ± 0.92 kg/d) and protein (1,028, 1,053, and 1,075 ± 28.7 g/d) were unaffected by level of RPM. However, the true protein content in milk from primiparous cows increased linearly (2.92, 3.09, and 3.34 ± 0.077%). The addition of RPM linearly increased the net flux of Met across the portal-drained viscera, which resulted in increased arterial Met concentrations (25, 29, and 40 ± 1.1 μM). Although it had no significant effect on net portal and hepatic fluxes of other essential amino acids, RPM resulted in a linear increase in the total splanchnic output of Ile, Leu, Phe, and Thr. These results suggest that feeding RPM triggered a homeostatic response resulting in less utilization of certain essential amino acids through the gastrointestinal tract and liver. Net mammary uptake of Met did not change with the addition of RPM. However, mammary extraction of Met decreased in a linear fashion in response to increased arterial inflow.     

Amino acid limitation and flow to the duodenum at four stages of lactation. 2. Extent of lysine limitation.

Four multiparous Holstein cows with ruminal and duodenal cannulas were assigned to 4 x 4 Latin squares at peak (wk 4), early (wk 14 to 16), mid (wk 21 to 23), and late (wk 29 to 31) lactation to determine, in the presence of supplemental Met, the extent of Lys limitation and its required contribution to total essential AA in duodenal digesta. Treatments were duodenal infusions of 1) water alone or water with 2) 10 g/d of DL-Met plus 10 g/d of L-Lys, 3) 10 g/d of Met plus 20 g/d of Lys, and 4) 10 g/d of Met plus 30 g/d of Lys; quantities were reduced by 20% in late lactation. Rations were corn based (corn and grass-legume silages, corn meal, wheat middlings, soybean meal, and distillers dried grains with solubles) and most limiting in Lys and Met. Intakes of ruminally degraded and undegraded intake protein (percentage of NRC requirements) were (peak) 115, 97; (early) 112, 83; (mid) 113, 87; and (late) 127, 96. Contribution of Lys to passage of total essential AA to the duodenum without infusions were 13.2, 12.4, 13.8, and 14.8% at the four respective stages of lactation. Extent of Lys limitation determined from responses in content and yield of milk protein approximated 25, 20, and 10 g/d during peak, early, and midlactation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Corn distillers grains versus a blend of protein supplements with or without ruminally protected amino acids for lactating cows

In a replicated 4 x 4 Latin square design with 4-wk periods, we used 12 multiparous Holstein cows averaging 83 d postpartum to compare corn distillers grains (CDG) versus a blend (BLEND) of other protein sources with CDG (fish meal and soybean meal), and to determine the effectiveness of ruminally protected lysine and methionine (RPLM) in improving the utilization of CDG as a protein supplement for lactating cows. The 2 x 2 factorial arrangement of treatments was as follows: CDG diet, CDG diet plus RPLM, BLEND diet, and BLEND diet plus RPLM. All diets contained 30% corn silage, 20% alfalfa hay, and 50% the respective corn-based concentrate mixture. The array of amino acids available for absorption when cows were fed the BLEND diet was more desirable than for the CDG diet according to Milk Protein Score and Cornell Net Carbohydrate and Protein System. Dry matter intakes were similar among all diets. Milk yields (32.6, 31.7, 32.8, and 32.8 kg/d, respectively) were similar for cows fed all diets. Milk fat yields and percentages (3.72, 3.76, 3.67, and 3.63%) were unaffected by diet, but milk protein percentages (3.23, 3.26, 3.25, and 3.26%) tended to be higher when fed RPLM. Concentrations of most protein fractions in milk were similar for all diets, although beta-lactoglobulin was increased slightly when cows were fed BLEND diets. Lysine, Met, and Phe were indicated as the most limiting amino acids for all diets according to extraction efficiency and transfer efficiency of amino acid from blood by the mammary gland. Methionine status was apparently improved by RPLM supplementation; Lys status was improved by the BLEND diets. Milk yield and composition when cows were fed CDG were not further improved by feeding blends of protein sources or RPLM; however, such dietary changes improved Lys and Met status of the cows.     

Effect of supply of metabolizable protein on splanchnic fluxes of nutrients and hormones in lactating dairy cows

The effect of the supply of metabolizable protein on splanchnic fluxes of nutrients and hormones was measured in six catheterized late-lactation Holstein cows in a crossover design. Two isonitrogenous diets (16.3% CP), but differing in rumen protein degradability and estimated metabolizable protein (MP) supply (1654 g/ d, Lo-MP; 1930 g/d, Hi-MP) were fed, each over a 35-d experimental period. On d 34 or 35, net fluxes of nutrients and hormones across the portal-drained viscera, the liver, and total splanchnic tissues were determined. Portal absorption of total, essential, nonessential, and branched-chain amino acids (AA) increased with the Hi-MP diet. Approximately 76% of the additional metabolizable protein supply was recovered as extra AA-N absorption in the portal vein. Liver removal of AA was not different between diets, and this resulted in a greater net release across the splanchnic tissues for the Hi-MP diet. This extra AA supply provided substrates for the observed increased milk protein yield for the Hi-MP diet. Fractional efficiencies of conversion of absorbed individual essential AA into milk protein ranged from 0.42 to 0.68. The corresponding efficiencies for utilization of postsplanchnic AA supply were 0.42 to 1.80. Provision of methionine, phenylalanine, and histidine beyond the liver were similar to outputs in milk protein but the other essential AA were supplied to peripheral tissues in excess of milk output, indicative of oxidative mechanisms in nonhepatic tissues. Net fluxes of glucose, NH3-N, and urea were not affected by the diets. Neither arterial concentrations of insulin, somatotropin, or IGF-1, nor net transfers across the portal-drained viscera or liver of insulin, were affected by the diets. Although portal release of glucagon was not different between the diets, a smaller proportion was removed by the liver on the Hi-MP diet. Metabolism of AA across the splanchnic tissue bed is a major determinant of the quantity and the profile of AA delivered to peripheral tissues.     

Production and nitrogen utilization in lactating dairy cows fed ground field peas with or without ruminally protected lysine and methionine

Previous research has shown that cows fed ≥24% of the diet dry matter (DM) as field peas decreased milk yield as well as concentration and yield of milk protein, possibly due to reduced DM intake and limited supply of Lys and Met. Twelve multiparous and 4 primiparous lactating Holstein cows were randomly assigned to 1 of 4 diets in a replicated 4 × 4 Latin square design. The diets contained (DM basis) 34.8% corn silage, 15.2% grass-legume silage, 5.9% roasted soybean, 2.4% mineral-vitamin premix, 2.0% alfalfa pellets, and either (1) 36% ground corn, 2.4% soybean meal, and 1.3% urea (UR), (2) 29.7% ground corn, 9.8% soybean meal, 0.13% ruminally protected (RP) Lys, and 0.07% RP-Met (CSBAA), (3) 25% ground field peas, 12.3% ground corn, and 2.4% soybean meal (FP), or (4) FP supplemented with 0.15% RP-Lys and 0.05% RP-Met (FPAA). Our objective was to test the effects of FP versus UR, FPAA versus CSBAA, and FPAA versus FP on milk yield and composition, N utilization, nutrient digestibility, ruminal fermentation profile, and plasma concentration of AA. Milk yield did not differ across diets. Compared with cows fed UR, those fed FP had greater DM intake, concentration and yield of milk true protein, apparent total-tract digestibility of fiber, urinary excretion of purine derivatives, and concentrations of total volatile fatty acids in the rumen and Lys in plasma, and less milk urea N and ruminal NH₃-N. The concentration of milk urea N, as well as the concentration and yield of milk fat increased in cows fed FPAA versus CSBAA. Moreover, cows fed FPAA had greater ruminal concentration of total volatile fatty acids, increased proportions of acetate and isobutyrate, and decreased proportions of propionate and valerate than those fed CSBAA. The plasma concentrations of His, Leu, and Phe decreased, whereas plasma Met increased and plasma Lys tended to increase in cows fed FPAA versus CSBAA. Concentration of milk true protein, but not yield, was increased in cows fed FPAA versus FP. However, cows fed FPAA showed decreased concentrations of His and Leu in plasma compared with those fed FP. Overall, compared with the CSBAA diet, feeding FPAA did not negatively affect milk yield and milk protein synthesis. Furthermore, RP-Lys and RP-Met supplementation of the FP diet did not improve milk yield or milk protein synthesis, but decreased urinary urea N excretion.     

Microbial composition and omasal flows of bacterial,protozoal, and nonmicrobial amino acids in lactating dairy cows fed fresh perennial ryegrass (Lolium perenne L.) not supplemented or supplemented with rolled barley

The objective of this study was to evaluate the effect of rolled barley supplementation on microbial composition and omasal flows of bacterial, protozoal, and nonmicrobial AA in cows fed fresh perennial ryegrass (Lolium perenne L.; PRG). Ten ruminally cannulated multiparous Holstein cows averaging (mean ± standard deviation) 49 ± 23 d in milk and 513 ± 36 kg of body weight were assigned to 1 of 2 treatments in a switchback design. The treatment diets were PRG only or PRG plus 3.5 kg of dry matter rolled barley (G+RB). The study consisted of three 29-d periods where each period consisted of 21 d of diet adaptation and 8 d of data and sample collection. A double-marker system was used to quantify nutrient flow entering the omasal canal along with ¹⁵N-ammonium sulfate to label and measure the microbial and nonmicrobial omasal flow of AA. Overall, rolled barley supplementation had no effect on the AA composition of the omasal liquid-associated and particle-associated bacteria. Rolled barley supplementation affected the AA concentrations of omasal protozoa; however, the differences were nutritionally minor. Particle-associated bacteria AA flow was increased for all AA, except for Trp and Pro, in cows fed the G+RB diet. Rolled barley supplementation had no effect on protozoal AA flow. On average, protozoa accounted for 23% of the microbial essential AA flow, which ranged from 17 to 28% for Trp and Lys, respectively. The flow of all AA in omasal true digesta increased in cows fed the G+RB diet compared with the PRG-only diet, resulting in a 228 g/d increase in total AA flow in cows fed the G+RB diet. This increase in total AA flow in cows fed the G+RB diet was due to an increase in microbial AA flow. Rolled barley supplementation had no effect on nonmicrobial AA flow. The nonmicrobial AA flow modestly contributed to total AA flow, accounting for 15.6% on average. These results indicated that extensive ruminal degradation of PRG AA occurred (83.5%), and we demonstrated that cows consuming PRG-based diets exhibit a large dependence on microbial AA to support metabolizable AA supply. Rolled barley supplementation can increase the omasal flow of microbial AA in cows consuming PRG-based diets. However, further research is required to elucidate if this increased AA supply can support higher milk yield under such dietary conditions.     

Effect of supply of metabolizable protein on whole body and splanchnic leucine metabolism in lactating dairy cows

The effect of the supply of metabolizable protein (MP) on protein metabolism across the splanchnic tissues was determined in six catheterized lactating Holstein cows. In a crossover design, two isonitrogenous (16.3% CP) diets balanced to provide a low (Lo-MP) or high (Hi-MP) supply of MP were fed over 35-d periods. After 24 d of feeding, N balance was determined over a 6-d period. On d 33, [13C] sodium bicarbonate was infused into one jugular vein for 6 h, and hourly breath samples were collected. On d 34 or 35, L[1-(13)C] leucine was infused into one jugular vein, and between 2 to 6 h of infusion, breath and blood samples were taken hourly from the portal and hepatic veins and an artery. Isotopic enrichments of plasma leucine, 4-methyl-2-oxopentanoate, and expired CO2 were determined for calculation of leucine kinetics. Net leucine absorption was greater, either on a direct basis (leucine transfer only) or corrected for portal-drained viscera metabolism to 4-methyl-2-oxopentanoate and CO2 for the Hi-MP diet. There were no effects of diet on hepatic net flux of leucine across the liver, and, thus, more leucine was available to peripheral tissues with the Hi-MP diet. Combined with an increment in portal absorption of most of essential AA, this led to increased milk protein output, although it only represented 16% of the additional available leucine. Whole body leucine oxidation was also greater for the Hi-MP diet, as was leucine used for protein synthesis. Despite these changes, MP supply did not affect irreversible loss rate of leucine by portal-drained viscera and the liver; these averaged 35 and 20% of whole body irreversible loss rate, respectively. These ratios confirm the high metabolic activity of splanchnic tissues in lactating dairy cows, which are even greater than previously reported in growing ruminants.