Abomasal Infusion Of Casein Research Articles

Effect of Post-Ruminal Casein Infusion on Milk Yield, Milk Composition, and Efficiency of Nitrogen Use in Dairy Cows

Adequate supply of amino acids can improve the efficiency of nitrogen use. Casein is the predominant milk protein, and its supplementation can improve milk protein synthesis and nitrogen efficiency. We evaluated the effects of post-ruminal supplementation of casein on milk yield and composition and whole-body protein deposition. Two ruminally cannulated Holstein dairy cows (599 kg) were used in a switch-back design, and treatments were an abomasal infusion of 0 or 400 g/day casein. Cows were fed a diet consisting of corn silage, alfalfa hay, wet corn gluten feed, whole cottonseed, and grain mix, and they received 320 g/day dextrose via abomasal infusion to increase energy:metabolizable protein. The experiment used three 8-day periods. Milk, urine, and feces samples were collected to evaluate milk production, milk composition, and nitrogen retention. Abomasal casein infusion increased (p < 0.01) milk protein percentage and milk urea nitrogen. Nitrogen retention (p = 0.03) and urinary N excretion (p < 0.001) were increased and fecal N excretion (p < 0.001) was decreased by casein infusion. Results suggest casein stimulated protein deposition and altered nitrogen use in lactating dairy cattle. Adaptation periods of 4 days were appropriate for evaluating responses to casein supplementation. Our data provide elements that can aid the design of future experiments.

Effect of experimentally increased protein supply to postpartum dairy cows on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis.

The effect of experimentally increasing the postpartum protein supply on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis was studied using 8 periparturient Holstein cows in a complete randomized design. At calving, cows were assigned to abomasal infusion of water (CTRL) or casein (CAS) in addition to a lactation diet. Casein infusion was gradually decreased from 696 ± 1 g/d at +2 d relative to calving (DRTC) to 212 ± 10 g/d at +29 DRTC to avoid excessive supply. Synthesis rate of plasma proteins was measured at -14, +4, +15, and +29 DRTC by measuring [C]Phe isotopic enrichment in arterial plasma free Phe, total plasma proteins, and albumin after 3, 5, and 7 h of jugular ring[C]Phe infusion. Plasma volume was determined at +4 and +29 DRTC by dilution of a [I]BSA dose. Synthesis rate of tissue protein in biopsied rumen papillae was determined by measuring [C]Phe isotopic enrichment, and mRNA expression of selected genes was measured by real-time qPCR. Total and differential leukocyte counts were performed and immune responsiveness of monocytes was evaluated by tumor necrosis factor ɑ (TNFɑ) concentration on ex vivo whole blood stimulation with Escherichia coli lipopolysaccharide (LPS) and responsiveness of T-lymphocytes by interferon γ (IFNγ) concentration on stimulation with Staphylococcus aureus enterotoxin β (SEB). Further, ELISA plasma concentrations of IgM, IgA, and IgG were determined. The DRTC affected the majority of investigated parameters as expected. The CAS treatment increased milk protein yield (P = 0.04), and tended to lower TNFɑ (P = 0.06), and lowered IFNγ (P = 0.03) responsiveness per monocyte and lymphocyte, respectively, compared with CTRL. Further, fractional synthesis rate of albumin was greater at +4 DRTC for CAS compared with CTRL but did not differ by +29 DRTC (interaction: P = 0.01). In rumen papillae, synthesis rate of tissue protein was greater for CAS compared with CTRL (P < 0.01) and mRNA expression of genes for cell proliferation tended to be or were greater for CAS compared with CTRL (P ≤ 0.07). In conclusion, increased postpartum protein supply seem to enhance vital body functions as interpreted from increased liver synthesis of albumin, increased rumen papillae proliferation, and stabilized the ex vivo inflammatory responsiveness of leukocytes. Further studies are needed to enlighten the importance of increased postpartum protein supply in periparturient cows.

1516 Lipogenic gene network expression in mammary tissue in response to abomasal infusion of casein, glucose, and acetate into feed-restricted lactating cows

1516 Lipogenic gene network expression in mammary tissue in response to abomasal infusion of casein, glucose, and acetate into feed-restricted lactating cows

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.

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

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

Effects of glucose and amino acids on ghrelin secretion in sheep

Two experiments were conducted to elucidate the effects of post-ruminal administration of starch and casein (Exp. 1), plasma amino acids concentrations (Exp. 2), and plasma glucose and insulin concentrations (Exp. 2) on plasma ghrelin concentrations in sheep. In Exp. 1, plasma ghrelin concentrations were determined by four infusion treatments (water, cornstarch, casein and cornstarch plus casein) in four wethers. Abomasal infusion of casein increased plasma alpha-amino N (AAN) concentrations. Infusion of starch or casein alone did not affect plasma ghrelin concentrations, but starch plus casein infusion increased plasma levels of ghrelin, glucose and AAN. In Exp 2, we investigated the effects of saline or amino acids on ghrelin secretion in four wethers. Two hours after the initiation of saline or amino acid infusion into the jugular vein, glucose was also continuously infused to investigate the effects of blood glucose and insulin by hyper-glycemic clump on plasma ghrelin concentrations. Infusion of amino acids alone raised plasma levels of ghrelin, but the higher plasma glucose and insulin concentrations had no effect on plasma ghrelin concentrations. These results suggest that high plasma levels of amino acids can stimulate ghrelin secretion, but glucose and insulin do not affect ghrelin secretion in sheep.

Abomasal infusion of casein, starch and soybean oil differentially affect plasma concentrations of gut peptides and feed intake in lactating dairy cows

The effects of specific nutrients on secretion and plasma concentrations of gut peptides (glucagon-like peptide-1 (7–36) amide (GLP-1), glucose-dependent insulinotropic polypeptide (GIP), and cholecystokinin-8 (CCK)) differ across species, but are not reported for cattle. Our objective was to determine acute (hours) and chronic (1 week) effects of increased abomasal supply of protein, carbohydrate, or fat to the small intestine on dry matter intake (DMI) and plasma concentrations of GLP-1, GIP, CCK, and insulin. Four mid-lactation Holstein cows were used in a 4 × 4 Latin square design experiment. Treatments were 7-day abomasal infusions of water, soybean oil (500 g/d), corn starch (1100 g/d), or casein (800 g/d). Jugular vein plasma was obtained over 7 h at the end of the first and last day of infusions. Oil infusion decreased DMI on day 7, but total metabolizable energy (ME) supply (diet plus infusate) did not differ from water infusion. Casein and starch infusion had no effect on feed DMI; thus, ME supply increased. Decreased DMI on day 7 of oil infusion was accompanied by increased plasma GLP-1 concentration, but decreased plasma CCK concentration. Increased plasma GIP concentration was associated with increased ME supply on day 7 of casein and starch infusion. Casein infusion tended to increase plasma CCK concentration on both days of sampling, and increased plasma GLP-1 and insulin concentration on day 1 of infusion. The present data indicate a sustained elevation of plasma concentration of GLP-1, but not CCK, may contribute to the reduced DMI observed in dairy cows provided supplemental fat.

Responses of milk protein, arterial AA concentration, and mammary AA metabolism to graded abomasal casein infusion in lactating goats

It has been generally assumed in postruminal protein or AA infusion studies that casein or AA mixtures mimicking the essential AA (EAA) composition of milk approach ‘ideal protein’ status, but the differences in postabsorptive metabolism of individual EAA may bring the assumption into question. Graded amounts of caseinate sodium were abomasally infused into lactating goats, and concentrations in the plasma of carotid, mammary uptakes and utilization efficiencies of amino acids were monitored in the present study. The results indicated that arterial concentrations of all the amino acids, except for Arg were increased with the increase of caseinate sodium infused ( P < 0.05). The increments in plasma concentration differed with individual EAA. Concentrations of Ile, Leu, and Lys increased about two-folds, and that of Met, Thr, Phe, and His increased 58.33, 32.5, 29.87, and 61.66%, respectively, when data of the 99 g/day and the control treatment was compared. Mammary extraction efficiencies of Arg, Thr, Lys, Leu, and Phe increased with graded casein infusion ( P < 0.05), while those of the other EAA were not significantly changed ( P > 0.05). Mammary extraction efficiencies of individual non-essential amino acids (NEAA) varied greatly, and ranged from 9.18 to 33.89% in the control treatment. Except for Pro, mammary extraction efficiency of NEAA increased with graded casein infusion. Mammary uptake to milk output ratios ( U/ O) of sequestered AA was generally increased with the increase of casein infused, but that of Thr, His, and Ala was not insignificantly changed ( P > 0.05). Compared with the data of previous researchers, the U/ O of Thr recorded in the present study was much lower (<0.42). We speculate that Thr may be extracted in forms other than FAA (such as peptide bond AA) by the mammary under conditions of the present study. Consistent with the results of other authors, mammary uptake of Arg was two times more than milk output even in the control treatment, which suggests that there may be a requirement of Arg other than milk protein synthesis in the mammary.

Pancreatic exocrine secretion and plasma concentration of some gastrointestinal hormones in response to abomasal infusion of starch hydrolyzate and/or casein1

Eight Angus steers (290 +/- 8 kg), surgically prepared with pancreatic pouch-duodenal reentrant cannulas and abomasal infusion catheters were used in a replicated 4 x 4 Latin square experiment to investigate the effects of abomasal infusion of starch hydrolyzate (SH) and/or casein on pancreatic exocrine secretion and plasma concentration of hormones. Steers were fed a basal diet of alfalfa (1.2 x NEm) in 12 equal portions daily. Abomasal infusion treatments (6-L total volume infused per day) were water (control), SH [2.7 g/(kg BW x d)], casein [0.6 g/(kg BW x d)], and SH + casein. Periods were 3 d for adaptation and 8 d of full infusion. Pancreatic juice and jugular blood samples were collected over 30-min intervals for 6 h on d 11. Weight and pH of pancreatic samples were measured, and a 10% subsample was composited and frozen until analysis of total protein and pancreatic enzyme activities. The remaining sample was returned to the duodenum. Plasma was harvested and frozen until analyzed. Pancreatic juice (67 mL/h) and protein (1.8 g/h) secretion rates were not affected by nutrient infusion. There were SH x casein interactions for all pancreatic enzyme secretions (U/h; alpha-amylase, P < 0.03; trypsin, P < 0.08; and chymotrypsin, P < 0.03) and plasma insulin concentration (P < 0.10). Secretion of pancreatic enzymes was increased by SH (trypsin) and casein (alpha-amylase, trypsin, and chymotrypsin) but not when SH + casein were infused together. Glucose (P < 0.10) and cholecystokinin octapeptide concentrations (CCK-8; P < 0.05) were increased by SH, but glucagon was decreased (P < 0.10). Casein decreased (P < 0.10) plasma CCK-8 concentrations. These data indicate that positive effects of postruminal casein on enzyme secretion were inhibited by SH, emphasizing the complexity of the regulatory mechanisms involved in dietary adaptation of pancreatic exocrine secretion. Changes in hormone concentration may not relate directly to changes in enzyme secretion.

Nitrogen balance in lambs fed a high-concentrate diet and infused with differing proportions of casein in the rumen and abomasum.

Twenty-five wether lambs (34 +/- 0.9 kg) fitted with ruminal and abomasal infusion catheters were used in a completely randomized design to determine the effects of differing proportions of ruminal and abomasal casein infusion on N balance in lambs fed a high-concentrate diet (85% corn grain, 1.6% N; DM basis) for ad libitum intake. Wethers were infused with 0 (control) or 10.4 g/d of N from casein with ruminal:abomasal infusion ratios of 100:0, 67:33, 33:67, or 0:100% over a 14-d period. Feed, orts, feces, and urine were collected over the last 5 d. Total N intake and excretion were greater (P < 0.01) in lambs infused with casein than in controls; however, N retention did not differ in lambs infused with casein compared with controls, suggesting that N requirements were met without casein supplementation. Total N intake and total N excretion did not differ among casein infusion treatments. Urinary N excretion decreased linearly (P = 0.07) with decreasing ruminal infusion of casein. Site of casein infusion quadratically (P = 0.06) influenced N retained (g/d), with the greatest retention observed in the 33:67 ruminal:abomasal infusion treatment. Dry matter intake from feed decreased from 1,183 to 945 g/d (P = 0.02) in lambs infused with casein compared with controls, but apparently digested DM did not differ among treatments. These data indicate that decreasing the ruminal degradability of supplemental protein above that required to maximize N retention results in decreased urinary excretion of N without greatly affecting apparent diet digestion.

Nitrogen balance in lambs fed low-quality brome hay and infused with differing proportions of casein in the rumen and abomasum12

Twenty wether lambs (46 +/- 2 kg) fitted with ruminal and abomasal infusion catheters were used in a completely randomized design to determine the effects of differing proportions of ruminal and abomasal casein infusion on N balance in lambs fed low-quality brome hay (0.8% N, DM basis) for ad libitum intake. Wethers were infused with 0 (control) or 10.7 g/d of N from casein with ratios of ruminal:abomasal infusion of 100:0 (100R:0A), 67:33 (67R:33A), 33:67 (33R:67A), or 0:100% (0R:100A), respectively, over a 12-d period. Total N supply (hay N intake + N from casein infusion) was greater (P = 0.001) in lambs receiving casein infusion than in controls. Urinary N excretion (g/d) was greater (P = 0.001) in lambs receiving casein infusion than in controls. Urinary N excretion decreased as casein infusion was shifted from 100R:0A to 33R:67A and then slightly increased in lambs receiving 0R:100A (quadratic, P = 0.02). Total N excretion was greater (P = 0.001) in lambs receiving casein infusion than in controls and decreased linearly (P = 0.005) as casein infusion was shifted to the abomasum. Retained N (g/d, % of N intake, and % of digested N) was greater (P = 0.001) in lambs receiving casein than in controls. Retained N increased as infusion was shifted from 100R:0A to 33R:67A and then slightly decreased in lambs receiving 0R: 100A (quadratic, P < 0.07). Based on regression analysis, the predicted optimum proportion of casein infusion to maximize N retention was 68% into the abomasum. The regression suggests that supplementation with undegradable intake protein had an additional benefit over supplementation with ruminally degradable intake protein (100R:0A) and that changing the percentage of ruminally undegradable intake protein in supplemental protein from 33 to 100% resulted in minimal differences in N retention. Apparent N, DM, OM, and energy digestibility (% of intake) was greater (P < 0.03) in lambs infused with casein than controls but did not differ among casein infusion groups. These data suggest that feeding protein supplements containing a portion (greater than 0%) of the crude protein as ruminally undegradable intake protein, as compared to 100% ruminally degradable intake protein, to lambs consuming low-quality forage increases N retention and the efficiency of N utilization without influencing total-tract nutrient digestion.

Nitrogen balance in lambs fed a high-concentrate diet and infused with differing proportions of casein in the rumen and abomasum12

Twenty-five wether lambs (34 ± 0.9 kg) fitted with ruminal and abomasal infusion catheters were used in a completely randomized design to determine the effects of differing proportions of ruminal and abomasal casein infusion on N balance in lambs fed a high-concentrate diet (85% corn grain, 1.6% N; DM basis) for ad libitum intake. Wethers were infused with 0 (control) or 10.4 g/d of N from casein with ruminal:abomasal infusion ratios of 100:0, 67:33, 33:67, or 0:100% over a 14-d period. Feed, orts, feces, and urine were collected over the last 5 d. Total N intake and excretion were greater (P < 0.01) in lambs infused with casein than in controls; however, N retention did not differ in lambs infused with casein compared with controls, suggesting that N requirements were met without casein supplementation. Total N intake and total N excretion did not differ among casein infusion treatments. Urinary N excretion decreased linearly (P = 0.07) with decreasing ruminal infusion of casein. Site of casein infusion quadratically (P = 0.06) influenced N retained (g/d), with the greatest retention observed in the 33:67 ruminal:abomasal infusion treatment. Dry matter intake from feed decreased from 1,183 to 945 g/d (P = 0.02) in lambs infused with casein compared with controls, but apparently digested DM did not differ among treatments. These data indicate that decreasing the ruminal degradability of supplemental protein above that required to maximize N retention results in decreased urinary excretion of N without greatly affecting apparent diet digestion.

Effects of Casein and Glucose on Responses of Cows Fed Diets Based on Restrictively Fermented Grass Silage

This study was conducted to investigate whether well fed dairy cows given restrictively fermented grass silage diet will respond to incremental glucose and amino acid supply at early stage of lactation. Four rumen-cannulated Finnish Ayrshire cows were used in a 4×4 Latin square experiment with 14-d periods. The cows were fed good quality restrictively fermented grass silage ensiled with a formic acid additive for ad libitum intake. A concentrate mixture consisting of barley (85%) and solvent extracted rapeseed meal (11.4%) was given at a rate of 9 kg/d. The four treatments were continuous abomasal infusions of water (control), casein 300g/d, glucose 300g/d, and casein 300g/d + glucose 300g/d. The infusions had only minor effects on feed intake, diet digestibility, or rumen fermentation pattern. Both casein and glucose infusions increased milk, protein and lactose yields the effects being partly additive on the combined infusion. Infused casein increased milk protein and urea as well as plasma urea concentrations. Both casein and glucose tended to increase plasma glucose concentration. Casein increased arterial plasma concentrations of essential amino acids (EAA), branched-chain AA (BCAA), and total AA (TAA). Both casein and glucose, although glucose usually less than casein, increased arteriovenous differences of EAA, nonessential AA, BCAA, and TAA. Extraction efficiencies of AA were higher for glucose than for casein. Mammary plasma flow was at highest on the control diet, but reduced owing to infused nutrients, the reduction being less with combined rather than separate infusions of casein and glucose. Based on the partly additive increases in milk production parameters and changes in plasma metabolites, it is suggested that glucose alone increased milk protein yield by sparing AA from hepatic utilization, while casein increased both supply of AA and glucose. It was concluded that cows at early stage of lactation fed diets comprising of restrictively fermented grass silage and a cereal-based concentrate suffer from both limited AA and glucose supplies.

Na+/glucose co-transporter abundance and activity in the small intestine of lambs: enhancement by abomasal infusion of casein

The purpose of the present study was to determine the effect of abomasal casein infusion on glucose uptake and abundance of the Na+/glucose co-transporter (SGLT1) 1 in the ovine small intestine. Lambs (body weight 35 (sem 1.0) kg) were surgically fitted with abomasal infusion catheters and were fed diets containing equal portions of wheat hay and cracked maize. Lambs were infused with either 500 g water/d or with 500 g water containing 35 g casein/d. The infusion period lasted 10 d, after which lambs were killed, exsanguinated and eviscerated. Brush border membrane vesicles (BBMV) were prepared using mucosa from different small intestinal regions. Intake and total tract digestibility of nutrients were similar between treatments and averaged 1134, 1142 and 486 g/d and 67, 70, and 94 % for DM, organic matter and non-structural carbohydrates respectively. Crude protein (Nx6.25) digestibility was 15 % greater in the casein-infused than control lambs. Glucose uptake to BBMV ranged from 101 to 337 pmol/mg protein per s along the small intestine and was greatest in the mid-section of the small intestine. In the mid-jejunum, glucose uptake was greater (P<0.07) in lambs infused with casein and averaged 120 pmol/mg protein per s compared with 68 pmol/mg protein per s in the control group. SGLT1 affinity was similar between treatments and averaged 104 microm in the different segments of the small intestine of lambs. However, lambs infused with casein exhibited similar values along the small intestine and affinity averaged 106 microm, while in the control group a greater affinity (85 microm) was measured in the mid-jejunum. SGLT1 protein abundance was correlated with glucose uptake in the BBMV in the casein-treated lambs, but not in the control group. These results suggest that glucose uptake along the small intestine of lambs is influenced by casein or its derivatives in the small intestine via SGLT1 affinity and activity at the brush border membrane, and that SGLT1 activity may be regulated by post-translational events affected by amino acids and peptides.

Influence of substrate and/or neurohormonal mimic on in vitro pancreatic enzyme release from calves postruminally infused with partially hydrolyzed starch and/or casein.

Our objectives were to determine the effects of neuroendocrine challenge and substrates on in vitro alpha-amylase and trypsin release in pancreatic tissue collected from Holstein calves (n = 24; 88 +/- 3 kg) abomasally infused for 10 d with tap water (control), partially hydrolyzed starch (SH; 4 g/[kg of BW x d]) and/ or casein (0.6 g/[kg of BW x d]). The caudal portion of the pancreas was removed, rinsed with ice-cold saline, cut into approximately 2 x 2-mm segments, and incubated in oxygenated Krebs Ringer bicarbonate buffer containing no substrate (control), glucose, amino acids, or VFA at 39 degrees C. After 60 min of incubation, neurohormonal mimics (none; control), carbachol (acetylcholine analog; 10 microM final), or caerulein (cholecystokinin mimic; 100 nM final) were added to the flasks and tissue was incubated for 60 min. Pancreatic tissue concentrations and in vitro release of alpha-amylase and trypsin decreased (P < 0.001) in calves abomasally infused with SH. Carbachol increased (P < 0.10) alpha-amylase and trypsin release in tissue collected from all calves. An effect of caerulein to increase alpha-amylase release (P < 0.10) was only observed with prior exposure to abomasal casein infusion in vivo or with simultaneous incubation with amino acids in vitro. Caerulein increased (P < 0.10) trypsin release in tissue collected from all calves except for those receiving SH + casein. Glucose decreased (P < 0.10) alpha-amylase release from pancreatic tissue collected from calves receiving abomasal control and casein treatments. Amino acids decreased (P < 0.10) alpha-amylase and trypsin release from pancreatic tissue collected from calves receiving the abomasal control treatment. Glucose, amino acids, and VFA decreased (P < 0.10) trypsin release from tissue collected from calves receiving abomasal SH. These data indicate that carbachol can stimulate pancreatic enzyme release in vitro. Caerulein, however, is only effective in stimulating in vitro pancreatic enzyme release in tissue from calves with an increased postruminal protein supply or in tissue incubated with amino acids. The results indicate that postruminal and local nutrients might be important in altering the responsiveness to a neuroendocrine challenge and could be an important regulatory event involved with dietary adaptation in ruminants.

Effect of abomasally infused casein on post-ruminal digestibility of total non-structural carbohydrates and milk yield and composition in dairy cows

AbstractA study was conducted to evaluate the effect of abomasal infusion of casein on post-ruminal digestibility of starch and on milk yield and composition. Six multiparous Israeli Holstein cows in mid lactation, fitted with ruminal and abomasal cannulas, were used in a 3 ✕ 3 Latin-square experiment. Each cow received 1600 g maize starch infused into the abomasum. Treatments were: abomasal infusion of sodium caseinate at 0, 350, or 700 g casein daily. Chromium mordant neutral-detergent fibre (NDF) was used as a digesta marker. Casein infusion was associated with increases in post-ruminal and total-tract digestibility of non-structural carbohydrates and protein. Concentrations of rumen ammonia and of plasma insulin, glucose and urea were higher in casein-infused cows. Concentrations of milk protein and lactose and milk protein yield increased with casein infusion. Results indicate that increased protein flow to the abomasum can improve the yield of milk constituents in dairy cows. Digestibility and yield responses to infusion of 350 g casein per day were similar to those at 700 g/day. There was a tendency toward reduced milk protein efficiency in casein-infused cows. It is suggested that part of the production response can be related to a direct effect of protein supply and the rest can be explained by the indirect effect of improved carbohydrate availability in the small intestine.

Evaluation of Whole Blood and Plasma in the Interorgan Supply of Free Amino Acids for the Mammary Gland of Lactating Dairy Cows

We investigated the contribution of plasma and red blood cells to amino acid (AA) supply for milk protein synthesis during a combination of treatments that included abomasal infusion of casein and AA and utilization of a hyperinsulinemic-euglycemic clamp. Treatments resulted in substantial differences in circulating concentrations of AA, mammary uptake of AA, and rates of milk protein synthesis. Arterial concentrations of all AA in plasma were highly correlated with that of whole blood. Concentrations of AA in red blood cells were either higher (Asn+Asp, Gly, His, Leu, Met, Orn, Ser, Tau, Thr, and Tyr), lower (Ala, Arg, Cit, Cys, Ile, and Val), or similar (Gln+Glu, Phe, and Pro) to that of plasma. Arteriovenous difference measurements demonstrated that interorgan transfer of AA to the mammary gland was primarily by plasma. There was little involvement of red blood cells except for small quantities of Leu, Met, and Thr to the mammary gland; this contribution was greatest for Met and accounted for 14% of the total mammary uptake. Countercurrent transport of Gln + Glu, Asn + Asp, and Pro was also evident where these AA were extracted from plasma, but were released into red blood cells as blood passed through the mammary gland. This net influx of Gln+Glu, Asn+Asp, and Pro into red blood cells was equivalent to 26, 17, and 30% of their mammary uptake from plasma. Overall, the interorgan transport of free AA for the mammary gland was predominantly by plasma, and red blood cells were limited to minor contri butions in mammary uptake for a few AA. Furthermore, arteriovenous differences of essential AA across the mammary gland were highly correlated between plasma and whole blood.

Effects of Insulin and Postruminal Supply of Protein on Use of Amino Acids by the Mammary Gland for Milk Protein Synthesis

We examined the relationships between amino acid supply, net utilization of amino acid by the mammary gland, and milk protein yield, in investigations that utilized a hyperinsulinemic-euglycemic clamp. A two-way crossed factorial design was employed. There were two 12-d periods involving abomasal infusions of either water or a mixture of casein (500 g/d) plus branched-chain amino acids (88 g/d), with a hyperinsulinemic-euglycemic clamp during the last 4 d of each period. During the clamp, insulin was infused at 1.0μg·kg BW–1 ·h–1 to increase circulating levels fourfold, and euglycemia was maintained by infusion of glucose. The insulin clamp treatments increased milk protein yield by 15 and 25% during abomasal infusion of water or casein plus branched-chain amino acids, respectively. Circulating concentrations of essential amino acids were reduced (33%) during insulin clamp treatments, especially branched-chain amino acids (41%). Arteriovenous difference of essential amino acids across the mammary gland was linearly related to their arterial concentrations. However, milk protein yield was not related to either arterial concentration or arteriovenous difference, for any of the essential amino acids. During insulin clamp treatments, the mammary gland was able to support the increased milk protein yields by increasing extraction efficiency of essential amino acids, mammary blood flow, and glucose uptake. Furthermore, a positive mammary balance of total amino nitrogen and carbon was maintained for all treatments. These adaptations demonstrate the unique ability of the mammary gland to adjust local conditions to allow for an adequate nutrient supply.

Effects of insulin and amino acids on milk protein concentration and yield from dairy cows.

Our study investigated the effect of insulin on the regulation of milk protein synthesis in well-fed cows (n = 4) with or without additional amino acids (AA). The design was a two-way crossed factorial with two 12-d periods involving abomasal infusions of either water or a mixture of casein (500 g/d) plus branched-chain AA (88 g/d). During the last 4 d of each period a hyperinsulinemic-euglycemic clamp was performed; insulin was infused at 1.0 microgram.kg of BW-1.h-1 to increase circulating levels fourfold, and euglycemia was maintained by infusion of glucose. Cows were fed a diet formulated to exceed requirements for metabolizable energy and protein. During abomasal water infusion, the insulin clamp increased milk protein yields by 15% (+128 g/d); when combined with abomasal infusion of casein plus branched-chain AA, milk protein yield was increased by 25% (+213 g/d). These increases resulted from equivalent increases in milk protein concentration and milk yield. Concentrations of casein and whey proteins in milk were increased by insulin clamp treatments; however, there were no major changes in the relative proportions of individual casein and whey proteins. Plasma concentrations of essential AA were reduced (-33%) during the insulin clamp treatments; effects were most dramatic for the branched-chain AA (-41%) and their keto acids (-45%). Results confirm the important regulatory role of the endocrine system in milk protein synthesis and demonstrate this potential to produce milk protein is not fully expressed.

Effects of increasing ruminally degraded nitrogen and abomasal casein infusion on net portal flux of nutrients in yearling heifers consuming a high-grain diet.

Seven Meat Animal Research Center (MARC) III heifers (410+/-25 kg) fitted with hepatic portal, mesenteric venous, carotid catheters, and an abomasal cannula were used in a 7 x 5 incomplete Latin square design experiment. The objective was to evaluate the effects of increasing levels of ruminally degradable N (RDN) with or without the addition of abomasally infused casein on portal-drained visceral (PDV) flux of nutrients. Treatments consisted of dietary CP percentage levels of 9.5 (control), control plus .72% dietary urea (11.5U), control plus 1.44% dietary urea (13.5U), control plus abomasally infused casein (250 g/d; 11.5C), or control plus .72% dietary urea and abomasally infused casein (250 g/d; 13.5UC). All diets contained (DM basis) 80% ground corn, 15% corn silage, and 5% dry supplement and were provided for ad libitum consumption. Nitrogen intake increased (linear, P < .001) as CP increased from 9.5 to 13.5%. Portal-drained visceral release of ammonia N increased (linear, P < .10) as RDN increased, and was greater (P < .05) when protein was fed compared with heifers fed control (P < .10). Urea N removal by PDV was not affected ( P > . 10) by level of RDN but was greatest when 11.5C was fed and least when 13.5UC was fed. Net alpha-amino N (AAN) release by PDV was greatest when 13.5UC was fed (309 mmol/h), least when 9.5% CP was fed (112 mmol/h), and intermediate for the other groups (205 to 252 mmol/h). These data suggest that removal of N by the PDV may promote microbial protein synthesis when dietary RDN is low. When RDN needs have been met and amino acids are deficient for the host, escape protein should be fed to increase amino acid absorption.