H. Lapierre

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 12 wk with 48 Holstein 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: -317 g/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 24.3 kg/d, respectively). Milk yield was decreased by DMP (35.2 kg/d), but was similar to ADMP (38.8 kg/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 ± 0.75 kg/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.

The route of absorbed nitrogen into milk protein

A database reviewing the metabolism of nitrogen (N) compounds from absorption to milk has been compiled from 14 published and unpublished studies (33 treatments) that measured the net flux of N compounds across the splanchnic tissues in dairy cows. Apparent N digestibility averaged 0·65, with this then partitioned between 0·34 excreted in urine and 0·31 secreted as milk. Nitrogen metabolites are absorbed from the lumen of the gut into the portal vein, mainly as free amino acids (AA) and ammonia; these represented 0·58 and 0·57 of digested N, respectively. All of the ammonia absorbed was removed by the liver with, as a result, a net splanchnic flux of zero. Detoxification of ammonia by the liver and catabolism of AA results in production of urea as an end-product. Hepatic ureagenesis is a major cross-road in terms of whole body N exchange, being the equivalent of 0·81 of digested N. Therefore, salvage of a considerable part of this ureagenesis is needed to support milk protein synthesis. This salvage occurs via transfer of urea from the blood circulation into the lumen of the gut. On average, 0·47 of hepatic ureagenesis was returned to the gut via the portal-drained viscera (equivalent to 0·34 of digested N) with 0·56 of this then used for anabolic purposes e.g. as precursor N for microbial protein synthesis. On average, 0·65 of estimated digestible AA was recovered in the portal vein. This loss (0·35) is due to oxidation of certain AA across the gut wall and non-absorption of endogenous secretions. The magnitude of this loss is not uniform among AA and varies between less than 0·05 for histidine to more than 0·90 for some non-essential AA, such as glutamine. A second database (six studies, 14 treatments) was constructed to further examine the subsequent fate of absorbed essential AA. When all AA are aggregated, the liver removed, on average, 0·45 of portal absorption but this value hides the considerable variation between individual AA. Simplistically, the AA behave as two major groups: one group undergoes very little hepatic removal and includes the branched-chain AA and lysine. For the second group, removal varies between 0·35 and 0·50 of portal absorption, and includes histidine, methionine and phenylalanine. For both groups, however, the efficiency of transfer of absorbed AA into milk protein decreases with increasing supply of protein. This loss of efficiency is linked directly with increased hepatic removal of AA from the second group and, probably, increased catabolism by peripheral tissues, including the mammary gland, of AA from the first group. Therefore, we must stop using fixed factors of conversion of digestible AA to milk in our predictive schemes and acknowledge that metabolism of AA between delivery from the duodenum and conversion to milk protein will vary with nutrient supply. New information evolving from re-analysis of the literature and recent studies will allow better models to be devised for the prediction of nutrient-based responses by the lactating cow. Consideration of biological efficiency, rather than maximal milk yield, will lead to systems that are economically more sensible for the farmer and that have better environmental impacts.

Alfalfa Cut at Sundown and Harvested as Baleage Improves Milk Yield of Late-Lactation Dairy Cows

Alfalfa (Medicago sativa L.) cut at sundown has been shown to contain greater concentration of total nonstructural carbohydrates (TNC) than that cut at sunup. Fourteen multiparous (8 ruminally cannulated) and 2 primiparous lactating dairy cows were randomly assigned to 2 treatments in a crossover design (2 periods of 24 d) to investigate the effects of alfalfa daytime cutting management on ruminal metabolism, nutrient digestibility, N balance, and milk yield. Half of each alfalfa field (total of 3 fields) was cut at sundown (PM) after a sunny day, whereas the second half was cut at sunup (AM) on the following day. Both PM and AM cuts were field-wilted and harvested as baleage (531 +/- 15.0 g of dry matter/kg of fresh matter). Bales (PM and AM) were ranked according to their concentrations of TNC, paired, and each pair of PM and AM baleages was then assigned to each experimental day (total of 48 d). The difference in TNC concentration between PM and AM baleages fed during the 10 d of data and sample collection varied from -10 to 50 g/kg of dry matter. Each pair of baleage was fed ad libitum to cows once daily with no concentrate. Ruminal molar proportion of acetate and total volatile fatty acid concentration were greater in animals fed the AM baleage, whereas the proportion of valerate was greater with PM baleage; no other significant changes in ruminal molar proportions of volatile fatty acids were observed between forage treatments. Digestible organic matter intake, organic matter digestibility, and plasma Lys concentration were significantly greater in cows fed PM alfalfa, suggesting that more nutrients were available for milk synthesis. Significantly lower body weight gain and retained N as a proportion of N intake were observed in cows fed PM alfalfa, thus suggesting that nutrients were channeled to milk synthesis rather than to body reserves. Intake of dry matter (+1.0 kg/d), and yields of milk (+1.0 kg/d), milk fat (+70 g/d), and milk protein (+40 g/d) were significantly greater in cows fed PM vs. AM alfalfa. Concentration of milk urea N and excretion of urea N as a proportion of total urinary N were significantly reduced, and milk N efficiency was increased when feeding PM vs. AM alfalfa, indicating an improvement in N utilization. Increasing the TNC concentration of alfalfa by shifting forage cutting from sunup to sundown improved N utilization and milk production in late-lactation dairy cows.

Effects of supplements of folic acid, vitamin B12, and rumen-protected methionine on whole body metabolism of methionine and glucose in lactating dairy cows.

The present experiment was undertaken to determine the effects of dietary supplements of rumen-protected methionine and intramuscular injections of folic acid and vitamin B(12), given 3 wk before to 16 wk after calving, on glucose and methionine metabolism of lactating dairy cows. Twenty-four multiparous Holstein cows were assigned to 6 blocks of 4 cows each according to their previous milk production. Within each block, 2 cows were fed a diet estimated to supply methionine as 1.83% metabolizable protein, equivalent to 76% of methionine requirement, whereas the 2 other cows were fed the same diet supplemented daily with 18 g of rumen-protected methionine. Within each diet, the cows were administrated either no vitamin supplement or weekly intramuscular injections of 160 mg of folic acid plus 10 mg of vitamin B(12.) To investigate metabolic changes at 12 wk of lactation, glucose and methionine kinetics were measured by isotope dilution using infusions of 3[U-(13)C]glucose, [(13)C]NaHCO(3) and 3[1-(13)C,(2)H(3)] methionine. Milk and plasma concentrations of folic acid and vitamin B(12) increased with vitamin injections. Supplementary B-vitamins increased milk production from 34.7 to 38.9 +/- 1.0 kg/d and increased milk lactose, protein, and total solids yields. Whole-body glucose flux tended to increase with vitamin supplementation with a similar quantitative magnitude as the milk lactose yield increase. Vitamin supplementation increased methionine utilization for protein synthesis through increased protein turnover when methionine was deficient and through decreased methionine oxidation when rumen-protected methionine was fed. Vitamin supplementation decreased plasma concentrations of homocysteine independently of rumen-protected methionine feeding, although no effect of vitamin supplementation was measured on methionine remethylation, but this could be due to the limitation of the technique used. Therefore, the effects of these B-vitamins on lactation performance were not mainly explained by methionine economy because of a more efficient methylneogenesis but were rather related to increased glucose availability and changes in methionine metabolism.

Alfalfa cut at sundown and harvested as baleage increases bacterial protein synthesis in late-lactation dairy cows.

Alfalfa (Medicago sativa L.) cut at sundown (p.m.) has been shown to have a greater concentration of total nonstructural carbohydrates (TNC) than when cut at sunup (a.m.). Eight ruminally cannulated Holstein cows that were part of a larger lactation trial were used in a crossover design (24-d periods) to investigate the effects of alfalfa cutting time on digestibility and omasal flow of nutrients. Alfalfa was cut at sundown or sunup, field-wilted, and harvested as baleage (530 +/- 15.0 g of dry matter/kg of fresh matter). The difference in TNC concentration between p.m. and a.m. alfalfa within each pair of bales fed daily during the 10 d of data and sample collection varied from -10 to 50 g/ kg of dry matter. Each pair of bales was fed for ad libitum intake to cows once daily with no concentrate. During the 3 d of omasal sampling, intake (+0.8 kg/d) and omasal flow of organic matter (OM; +0.42 kg/d) tended to be greater when cows were fed p.m. vs. a.m. alfalfa, but no differences were found for ruminal and postruminal digestion of this nutrient. Similarly, N apparently digested ruminally and postruminally did not differ when feeding p.m. vs. a.m. alfalfa. However, N truly digested in the rumen, as a proportion of N intake, was significantly greater in cows fed p.m. (79%) vs. a.m. alfalfa (74%), thus suggesting that longer wilting time of alfalfa cut at sundown increased forage proteolysis. Supply of rumen-degradable protein did not change (2,716 g/d) when averaged across treatments, whereas omasal flow of non-NH(3) nonbacterial N was significantly decreased (-29 g/d) when feeding p.m. vs. a.m. alfalfa. Omasal flow of total bacterial non-NH(3)-N (NAN) increased (+21 g/d) significantly when cows were fed p.m. vs. a.m. alfalfa possibly because bacteria from cows fed p.m. alfalfa captured significantly more NH(3) than those from cows fed a.m. alfalfa. Therefore, greater availability of fermentable energy as TNC appears to increase the capacity of microbes to uptake NH(3)-N and convert it to microbial protein. Enhanced OM intake can also explain the observed increase in bacterial protein synthesis with p.m. alfalfa. Efficiency of bacterial protein synthesis, expressed on a fermented OM basis or as grams of bacterial NAN per gram of rumen-degradable N, did not differ between p.m. and a.m. alfalfa. Conversely, bacterial efficiency, as grams of bacterial NAN per gram of N intake, was significantly increased when cows were fed p.m. baleage. No significant difference between forage treatments was found for the omasal flow of total AA from omasal true digesta, suggesting no benefit of daytime cutting management on the passage of total AA to the lower gastrointestinal tract. Enhancing energy intake and TNC concentration of alfalfa by shifting forage cutting from sunup to sundown increased protein synthesis and NH(3) uptake by ruminal bacteria indicating an improvement in N utilization.

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 24 g 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 76 g of RPLys/cow per day (AMPL); and (2) AMPL (74 g of RPLys/cow per day) supplemented with 24 g of RPMet/cow per day (AMPLM). Each experiment lasted for 10 wk (2-wk adaptation and 8-wk experimental periods) following a 2-wk covariate period (i.e., a total of 12 wk). 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 -666 g/d) in a linear manner: milk yield under-prediction [National Research Council (2001) MP-allowable milk yield, kg/d - actual milk yield, kg/d] = 0.0991 (±0.0905) + 0.0230 (±0.0003) × MP balance, g/d (R(2)=0.99).

Insulin-like growth factor-I concentration in Holstein female cattle: variations with age, stage of lactation and growth hormone-releasing factor administration

Plasma insulin-like growth factor-I (IGF-I) concentrations were monitored in Holstein females through different periods of their growth, lactation and after acute or chronic growth hormone-releasing factor (GRF) administration. Plasma samples were radioimmunoassayed using a human IGF-I antibody after a 24 hr incubation in a HCl(.1N)-glycine(.2M) buffer (pH 2). In a first study, IGF-I concentrations were measured in Holstein females of different ages and(or) stages of lactation (n = 6 per group). The IGF-I concentrations in newborn calves (102.0 +/- 11.3 ng/ml) markedly decreased (P less than .01) in 1 mo old animals (50.2 +/- 7.1 ng/ml), then increased (P less than .01) to 137.0 +/- 5.1 and 137.4 +/- 11.0 ng/ml in 6 and 10 mo old heifers, respectively. In dairy cows, IGF-I concentrations were low 24 hr post-partum (44.7 +/- 7.6 ng/ml) and then increased (P less than .05) to remain stable throughout lactation (91.3 +/- 4.9, 92.8 +/- 12.9, 96.1 +/- 7.6, 90.7 +/- 8.8 ng/ml at 2, 3, 6 and 9 mo of lactation, respectively). There was a further increase (P less than .05) to 113.7 +/- 3.1 ng/ml during the dry period. In a second trial, blood samples were collected from lactating dairy cows every 2 hr for 24 hr following a sc injection of saline (n = 4) or human (h) GRF (1-29)NH2 (10 micrograms/kg BW, n = 4). The IGF-I peak concentration was reached on average 10 hr after the GRF injection and was higher (P less than .01) in treated cows than in control cows (135.4 vs 86.9 +/- 16.2 ng/ml). In the last trial, daily sc injections of 10 micrograms of hGRF(1-29)NH2 per kg BW to dairy cows (252 days of lactation) for 57 days, which increased milk production by 14% (2 kg/day), also increased (P less than .01) IGF-I concentration: 127.1 +/- 5.3 and 118.0 +/- 1.6 vs 90.7 +/- 4.7 and 96.0 +/- 5.0 ng/ml on days 29 and 57 of treatment for treated (n = 9) and control (n = 8) cows, respectively. Thus, the IGF-I concentration in dairy cattle varies with age and stage of lactation, and is increased by GRF administration in lactating dairy cows.

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

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

Whole-body glucose metabolism and mammary energetic nutrient metabolism in lactating dairy cows receiving digestive infusions of casein and propionic acid

This study analyzed the effect of propionate (C3) and casein (CN) on whole-body and mammary metabolism of energetic nutrients. Three multiparous Holstein cows fitted with both duodenal and ruminal cannulas were used in 2 replicated Youden squares with 14-d periods. Effects of CN (743 g/d in the duodenum) and C3 (1,042 g/d in the rumen) infusions, either separately or in combination as supplements to a grass silage diet, were tested in a factorial arrangement. The control diet provided 97% of energy and protein requirements. Within each period, blood samples were taken (d 11) from the carotid artery and the right mammary vein to determine net uptake of energetic nutrients. Plasma blood flow was calculated using the Fick principle (based on Phe and Tyr). On d 13, [6,6-(2)H(2)]glucose was infused in the jugular vein to determine whole-body glucose rate of appearance (Ra) based on enrichments in arterial plasma. Both C3 and CN treatments increased whole-body Ra (17% and 13%, respectively) but only CN increased milk (18%) and lactose (14%) yields, suggesting no direct link between whole-body Ra and milk yield. When CN was infused alone, the apparent ratio of conversion of CN carbon into glucose carbon was 0.31 but, when allowance was made for the CN required to support the extra milk protein output, the ratio increased to 0.40, closer to the theoretical ratio (0.48). This may relate to the observed increases in arterial glucagon concentrations for CN alone. Conversely, the apparent conversion of infused C3 carbon alone to glucose was low (0.31). With C3, mammary plasma flow increased as did uptakes of lactate, Ala, and Glu whereas the uptake for beta-hydroxybutyrate (BHBA) decreased. Mammary net carbon balance suggested an increase with C3 treatment in glucose, lactate, Ala, and Glu oxidation within the mammary gland. Mammary glucose uptake did not increase with CN treatment, despite an increase in glucose arteriovenous difference and extraction rate, because plasma flow decreased (-17%). Whereas CN, alone or in combination with C3, increased both lactose and protein yields, only mammary AA (and BHBA in CN alone) uptake increased because plasma flow decreased (-17%). These data suggest that the observed variations of milk lactose yield (and other milk components) are linked to metabolic interchanges between several energetic nutrients at both the whole-body and mammary levels and are not explained by increases in whole-body glucose availability.

Influence of age and sex on basal secretion of growth hormone (GH) and on GH-induced release by porcine GH-releasing factor pGRF(1-29NH2) in growing pigs.

The aim of this study was to determine the effect of age and sex on basal secretory patterns of growth hormone (GH) and growth hormone-releasing factor (GRF) induced GH release. Eighteen pigs (9 castrated males and 9 females) were stimulated with pGRF(1-29)NH2 at 7,11,15,19 and 23 weeks of age. Blood samples were taken from each animal via jugular vein cannulae every 20 min, from 6 hr before to 5 hr after iv GRF administration at a dose of 4 micrograms/kg. GH baseline levels, amplitude of the GH peaks, area under the GH peaks and the overall mean of GH serum levels decreased (P less than .001) with age in both sexes. Age also had a marked effect on GRF-induced GH release: the amplitude of GH peaks and area under the GH peaks decreased (P less than .001) with age. The GH response to pGRF(1-29)NH2 varied considerably, depending on the timing of the episodic endogenous secretion of GH. An immediate response (less than 30 min) was observed when GRF was injected at the end of a trough period or at the beginning of a peak, but there was no immediate response when GRF was injected at the end of a peak or at the beginning of a trough period. Our results show that both endogenous GH secretion and pGRF(1-29)NH2-induced GH release declines with age, suggesting a decreased sensitivity of the somatotroph cells to GRF with age; and that the high variability of the GH response to pGRF(1-29)NH2 stimulation depends greatly on the timing of the episodic endogenous GH release, thus implying a possible episodic endogenous somatostatin secretion by the hypothalamus.