Ruminal ammonia load affects leucine utilization by growing steers

Six ruminally cannulated Holstein steers (initial BW = 189 +/- 11 kg) housed in metabolism crates were used in a 6 x 6 Latin square to study effects of ruminal ammonia load on Leu utilization. All steers received a diet based on soybean hulls (2.7 kg of DM/d), ruminal infusions of 200 g of acetate/d, 200 g of propionate/d, and 50 g of butyrate/d, as well as an abomasal infusion of 300 g of glucose/d to provide energy without increasing microbial protein supply and an abomasal infusion of a mixture (238 g/d) of all essential AA except Leu. Treatments were arranged as a 3 x 2 factorial and included Leu (0, 4, or 8 g/d) infused abomasally and urea (0 or 80 g/d) infused ruminally. Abomasal Leu infusion linearly decreased (P < 0.05) both urinary and fecal N excretions and linearly increased (P < 0.05) retained N, but the decreases in urinary N excretion in response to Leu tended (P = 0.07) to be greater, and the increases in retained N in response to Leu were numerically greater in the presence of the urea infusion. Although urea infusions increased (P < 0.05) plasma urea concentrations, urinary N excretions, and urinary urea excretions, retained N also was increased (P < 0.05). The efficiency of deposition of supplemental Leu ranged from 24 to 43% when steers received 0 or 80 g of urea/d, respectively. Under our experimental conditions, increasing ammonia load improved whole-body protein deposition in growing steers when Leu supply was limiting.     

Effects of energy source on methionine utilization by growing steers

We evaluated the effects of different supplemental energy sources on Met use in growing steers. Ruminally cannulated Holstein steers were used in two 6 x 6 Latin squares, and data were pooled for analyses. In Exp. 1, steers (148 kg) were fed 2.3 kg of DM/d of a diet based on soybean hulls. Treatments (2 x 3 factorial) were abomasal infusion of 0 or 3 g of l-Met/d, and supplementation with no energy or with glucose (360 g/d) or fat (150 g/d) continuously infused into the abomasum. In Exp. 2, steers (190 kg) received 2.6 kg of dietary DM/d and were provided (2 x 3 factorial) with 0 or 3 g of l-Met/d, and with no supplemental energy or with acetate (385 g/d) or propionate (270 g/ d) continuously infused into the rumen. In both experiments, the energy sources supplied 1.3 Mcal of GE/d, and all steers received basal infusions of 400 g of acetate/d into the rumen and a mixture (125 g/d) of all essential AA except Met into the abomasum. Nitrogen balance (18.8 vs. 23.5 g/d; P < 0.01) and whole-body protein synthesis (2.1 vs. 2.3 kg/d; P < 0.07) were increased by Met supplementation, indicating that protein deposition was limited by Met. Supplemental energy reduced (P < 0.01) urinary N excretion and increased (P < 0.01) N retention without differences among energy sources. Increases in N retention in response to Met were numerically greater when energy was supplemented. Efficiency of supplemental Met use was 11% when no energy was supplemented but averaged 21% when 1.3 Mcal of GE/d was provided. Whole-body protein synthesis and degradation were not affected by energy supplementation. Serum insulin concentrations were increased by glucose and propionate supplementation. Serum IGF-I concentrations were increased by supplementation with Met or glucogenic sources of energy. In growing steers, N retention was increased by energy supplementation even though protein deposition was limited by Met, suggesting that energy supplementation improves the efficiency of AA use. These responses were independent of the source of energy.     

Histidine utilization by growing steers is not negatively affected by increased supply of either ammonia or amino acids

Two experiments were conducted with ruminally cannulated Holstein steers to determine effects of N supply on histidine (His) utilization. All steers received 2.5 kg DM/d of a diet based on soybean hulls; abomasal infusion of 250 g/d amino acids, which supplied adequate amounts of all essential amino acids except His; abomasal infusion of 300 g/d glucose; and ruminal infusion of 180 g/d acetate, 180 g/d propionate, and 45 g/d butyrate. Both experiments were 6 x 6 Latin squares with treatments arranged as 3 x 2 factorials. No significant (P < 0.05) interactions between main effects were noted for N balance criteria in either Exp. 1 or 2. For Exp. 1, steers (146 +/- 7 kg) received 0, 1.5, or 3 g/d of L-His infused abomasally in combination with 0 or 80 g/d urea infused ruminally to supply a metabolic ammonia load. Urea infusions increased (P < 0.05) ruminal ammonia concentration from 8.6 to 19.7 mM and plasma urea from 2.7 to 5.1 mM. No change in N retention occurred in response to urea (35.1 and 37.1 g/d for 0 and 80 g/d urea, respectively, P = 0.16). Retained N increased linearly (P < 0.01) with His (31.5, 37.8, and 39.0 g/d for 0, 1.5, and 3 g/d L-His, respectively). Efficiency of deposition of supplemental His between 0 and 1.5 g/d averaged 65%. In Exp. 2, steers (150 +/- 6 kg) were infused abomasally with 0 or 1 g/d of L-His in combination with no additional amino acids (Control), 100 g/d of essential + 100 g/d of nonessential amino acids (NEAA+EAA), or 200 g/d of essential amino acids (EAA). Retained N increased (P = 0.02) from 34.2 to 38.3 g/d in response to His supplementation. Supplementation with NEAA+EAA increased (P < 0.05) N retention (33.9, 39.3, and 35.6 g/d for Control, NEAA+EAA, and EAA, respectively), likely in response to increased energy supply. Plasma urea concentrations of steers receiving NEAA+EAA (3.8 mM) and EAA (3.8 mM) were greater (P < 0.05) than those of Control steers (2.7 mM). The average efficiency of His utilization was 63%, a value similar to the value of 65% observed in Exp. 1, as well as the 71% value predicted by the Cornell net carbohydrate and protein system model. Under our experimental conditions, increases in N supply above requirements, as either ammonia or amino acids, did not demonstrate a metabolic cost in terms of His utilization for whole-body protein deposition by growing steers.     

Effects of energy level on methionine utilization by growing steers

We evaluated the effect of energy supplementation on Met use in growing steers. Six ruminally cannulated Holstein steers (228 +/- 8 kg of BW) were used in a 6 x 6 Latin square and fed 2.8 kg of DM/d of a diet based on soybean hulls. Treatments were abomasal infusion of 2 amounts of Met (0 or 3 g/d) and supplementation with 3 amounts of energy (0, 1.3, or 2.6 Mcal of GE/d) in a 2 x 3 factorial arrangement. The 1.3 Mcal/d treatment was supplied through ruminal infusion of 90 g/d of acetate, 90 g/d of propionate, and 30 g/d of butyrate, and abomasal infusion of 30 g/d of glucose and 30 g/d of fat. The 2.6 Mcal/d treatment supplied twice these amounts. All steers received basal infusions of 400 g/d of acetate into the rumen and a mixture (125 g/d) containing all essential AA except Met into the abomasum. No interactions between Met and energy levels were observed. Nitrogen balance was increased (P < 0.05) by Met supplementation from 23.6 to 27.8 g/d, indicating that protein deposition was limited by Met. Nitrogen retention increased linearly (P < 0.05) from 23.6 to 27.7 g/d with increased energy supply. Increased energy supply also linearly reduced (P < 0.05) urinary N excretion from 44.6 to 39.7 g/d and reduced plasma urea concentrations from 2.8 to 2.1 mM. Total tract apparent OM and NDF digestibilities were reduced linearly (P < 0.05) by energy supplementation, from 78.2 and 78.7% to 74.3 and 74.5%, respectively. Whole-body protein synthesis and degradation were not affected significantly by energy supplementation. Energy supplementation linearly increased (P < 0.05) serum IGF-I from 694 to 818 ng/mL and quadratically increased (P < 0.05) serum insulin (0.38, 0.47, and 0.42 ng/mL for 0, 1.3, and 2.6 Mcal/d, respectively). In growing steers, N retention was improved by energy supplementation, even when Met limited protein deposition, suggesting that energy supplementation affects the efficiency of AA use.     

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.     

Effects of ammonia load on methionine utilization by growing steers

Seven ruminally cannulated Holstein steers (194 +/- 16 kg) housed in metabolism crates were used in a 6 x 6 Latin square, with one additional steer, to study effects of ruminal ammonia load on methionine (Met) use. All steers received a diet based on soybean hulls (2.6 kg DM/d), ruminal infusions of 200 g/d of acetate, 200 g/d of propionate, and 50 g/d of butyrate, as well as abomasal infusion of 300 g/d of glucose to provide energy without increasing microbial protein supply, and abomasal infusions of a mixture (248 g/d) of all essential AA except Met. Treatments were arranged as a 3 x 2 factorial and included urea (0, 40, or 80 g/d) infused ruminally to supply metabolic ammonia loads and Met (2 or 5 g/d) infused abomasally. Supplementation with the greater amount of Met decreased (P < 0.05) urinary N excretion from 68.8 to 64.8 g/d and increased (P < 0.05) retained N from 22.0 to 27.5 g/d. Urea infusions linearly increased (P < 0.05) urinary N excretions, plasma urea concentrations, and urinary urea excretions, but retained N was not affected. The efficiency of deposition of supplemental Met, calculated by assuming that Met deposition is 2.0% of protein deposition (6.25 x retained N), ranged between 18 and 27% when steers received 0 or 80 g/d of urea, respectively. There were no (P > or = 0.40) effects of treatments on serum insulin or IGF-I concentrations. In our model, increasing ammonia load did not affect whole-body protein deposition in growing steers when Met was limiting.     

Effects of guanidinoacetic acid supplementation on nitrogen retention and methionine flux in cattle

Creatine stores high-energy phosphate bonds in muscle and is synthesized in the liver through methylation of guanidinoacetic acid (GAA). Supplementation of GAA may therefore increase methyl group requirements, and this may affect methyl group utilization. Our experiment evaluated the metabolic responses of growing cattle to postruminal supplementation of GAA, in a model where methionine (Met) was deficient, with and without Met supplementation. Seven ruminally cannulated Holstein steers (161 kg initial body weight [BW]) were limit-fed a soybean hull-based diet (2.7 kg/d dry matter) and received continuous abomasal infusions of an essential amino acid (AA) mixture devoid of Met to ensure that no AA besides Met limited animal performance. To provide energy without increasing the microbial protein supply, all steers received ruminal infusions of 200 g/d acetic acid, 200 g/d propionic acid, and 50 g/d butyric acid, as well as abomasal infusions of 300 g/d glucose. Treatments, provided abomasally, were arranged as a 2 × 3 factorial in a split-plot design, and included 0 or 6 g/d of l-Met and 0, 7.5, and 15 g/d of GAA. The experiment included six 10-d periods. Whole body Met flux was measured using continuous jugular infusion of 1-¹³C-l-Met and methyl-²H₃-l-Met. Nitrogen retention was elevated by Met supplementation (P < 0.01). Supplementation with GAA tended to increase N retention when it was supplemented along with Met, but not when it was supplemented without Met. Supplementing GAA linearly increased plasma concentrations of GAA and creatine (P < 0.001), but treatments did not affect urinary excretion of GAA, creatine, or creatinine. Supplementation with Met decreased plasma homocysteine (P < 0.01). Supplementation of GAA tended (P = 0.10) to increase plasma homocysteine when no Met was supplemented, but not when 6 g/d Met was provided. Protein synthesis and protein degradation were both increased by GAA supplementation when no Met was supplemented, but decreased by GAA supplementation when 6 g/d Met were provided. Loss of Met through transsulfuration was increased by Met supplementation, whereas synthesis of Met from remethylation of homocysteine was decreased by Met supplementation. No differences in transmethylation, transsulfuration, or remethylation reactions were observed in response to GAA supplementation. The administration of GAA, when methyl groups are not limiting, has the potential to improve lean tissue deposition and cattle growth.     

Branched-chain amino acids for growing cattle limit-fed soybean hull-based diets.

Five ruminally cannulated Holstein steers (176 kg) were used in a 5 x 5 Latin square to evaluate the effects of branched-chain AA supplementation on N retention and plasma AA concentrations of steers. Steers were limit-fed (3.0 kg/d of DM) twice daily diets low in ruminally undegradable protein (72% soybean hulls, 19% alfalfa, 5% molasses, and 4% vitamins and minerals). Acetate (400 g/d) was continuously infused into the rumen. Treatments were continuous abomasal infusions of 1) 115 g/d of a mixture of 10 essential AA designed to exceed the steers' requirements (10AA), 2) 10AA with Leu removed, 3) 10AA with Ile removed, 4) 10AA with Val removed, and 5) 10AA with all three branched-chain AA removed. Experimental periods were 7 d, with 3 d for adaptation to treatments and 4 d for total fecal and urinary collections for N balance. Blood samples were collected 5 h after feeding on d 7. Retained N decreased in response to removal of Leu (P < 0.06), Val (P < 0.05), or all three branched-chain AA (P < 0.05). Plasma Leu concentrations decreased (P < 0.05) in response to removal of Leu and all three branched-chain AA. Plasma Ile concentrations decreased (P < 0.05) in response to removal of Ile and all three branched-chain AA but increased (P < 0.05) in response to removal of Leu. Plasma Val concentrations decreased (P < 0.05) in response to removal of Val and all three branched-chain AA but increased (P < 0.05) in response to removal of Leu. Responses in N balance and plasma AA concentrations of growing cattle limit-fed soybean hull-based diets demonstrate limitations in the basal supply of Leu and Val but not Ile provided that supplies of all other essential AA are met.     

Excess amino acid supply improves methionine and leucine utilization by growing steers

In 2 experiments, 6 ruminally cannulated Holstein steers (205 +/- 23 and 161 +/- 14 kg initial BW in Exp. 1 and 2, respectively) housed in metabolism crates were used in 6 x 6 Latin squares to study the effects of excess AA supply on Met (Exp. 1) and Leu (Exp. 2) use. All steers received a diet based on soybean hulls (DMI = 2.66 and 2.45 kg/d in Exp. 1 and 2, respectively); ruminal infusions of 200 g of acetate/d, 200 g of propionate/d, and 50 g of butyrate/d, as well as abomasal infusion of 300 g of glucose/d to provide energy without increasing the microbial protein supply; and abomasal infusions of a mixture of all essential AA except Met (Exp. 1) or Leu (Exp. 2). Periods were 6 d, with 2-d adaptations and 4 d to collect N balance data. All treatments were abomasally infused. In Exp. 1, treatments were arranged as a 2 x 3 factorial, with 2 amounts of l-Met (0 or 4 g/d) and 3 AA supplements (no additional AA, control; 100 g/d of nonessential AA + 100 g/d of essential AA, NEAA + EAA; and 200 g/d of essential AA, EAA). Supplemental Met increased (P < 0.01) retained N and decreased (P < 0.01) urinary N and urinary urea N. Retained N increased (P < 0.01) with NEAA + EAA only when 4 g/d of Met was provided, but it increased (P < 0.01) with EAA with or without supplemental Met. Both AA treatments increased (P < 0.01) plasma urea and serum insulin. Plasma glucose decreased (P = 0.03) with supplemental Met. In Exp. 2, treatments were arranged as a 2 x 3 factorial with 2 amounts of L-Leu (0 or 4 g/d) and 3 AA supplements (control, NEAA + EAA, and EAA). Supplemental Leu increased (P < 0.01) retained N and decreased (P < 0.01) urinary N and urinary urea N. Both AA treatments increased (P < 0.01) retained N, and they also increased (P < 0.01) urinary N, urinary urea N, and plasma urea. Serum insulin increased (P = 0.06) with supplemental Leu and tended (P = 0.10) to increase with both AA treatments. Supplementation with excess AA improved Met and Leu use for protein deposition by growing cattle.     

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

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.     

Limiting amino acids for growing Holstein steers limit-fed soybean hull-based diets

Studies were conducted to determine limiting amino acids (AA) for cattle limit-fed soybean hull-based diets. Ruminally cannulated Holstein steers were maintained in metabolism crates, fed the same basal diet (73% soyhulls, 19% alfalfa, DM basis), and given the same intraruminal infusions (400 g/d acetate; to supply energy without increasing microbial protein supply). Treatments were infused abomasally. In Exp. 1, steers (200 kg) were provided 1) water, 2) 10 g/d of methionine (MET), or 3) a mixture of 10 essential AA (10AA). Nitrogen retention (13.7 g/d) was greatest (P < .05) for steers receiving 10AA. Steers receiving MET (7.9 g/d) had greater (P < .05) N retention than control steers (5.4 g/d). In Exp. 2, steers (200 kg) were provided 10AA or 10AA with L-Lys deleted from the mixture. Steers receiving 10AA tended (P < .09) to have greater N retention (19.0 g/d) than those receiving no lysine (16.3 g/d). In Exp. 3, steers (194 kg) were provided 10AA or 10AA with L-Thr deleted from the mixture. Nitrogen retention was not affected by removal of threonine. In Exp. 4, steers (152 kg) were provided 10AA or 10AA with L-His, L-Trp, L-Arg, L-Phe, or branched-chain AA (L-Leu, L-Ile, and L-Val) removed. Nitrogen retention was reduced (P < .05) by removal of either L-His or the branched-chain AA. For steers limit-fed soybean hull-based diets, methionine was first-limiting; histidine, at least one of the branched-chain AA, and possibly lysine were also limiting.     

Methionine as a methyl group donor in growing cattle

Holstein steers were used in two 5 x 5 Latin square experiments to evaluate the sparing of methionine by alternative sources of methyl groups (betaine and choline). Steers were housed in metabolism crates and limit-fed a soybean hull-based diet high in rumen degradable protein. To increase energy supply, ruminal infusions of volatile fatty acids and abomasal infusions of glucose were provided. An amino acid mixture, limiting in methionine, was infused abomasally to ensure that nonsulfur amino acids did not limit protein synthesis. Treatments for Exp. 1 were abomasal infusion of 1) water, 2) 2 g/d L-methionine, 3) 1.7 g/d L-cysteine, 4) 1.6 g/d betaine, and 5) 1.7 g/d L-cysteine + 1.6 g/d betaine. Treatments for Exp. 2 were abomasal infusion of 1) water, 2) 2 g/d L-methionine, 3) 8 g/d betaine, 4) 16 g/d betaine, and 5) 8 g/d choline. In both experiments, nitrogen retention increased in response to methionine (P < 0.05), demonstrating a deficiency of sulfur amino acids. Responses to cysteine, betaine, and choline were all small and not significant. The lack of response to cysteine indicates that the response to methionine was not due to transsulfuration to cysteine or that cysteine supply did not alter the flux of methionine through transsulfuration. The lack of response to betaine suggests that the steers' needs for methyl groups were met by the dietary conditions or that betaine was relatively inefficient in increasing the remethylation of homocysteine to methionine and, thereby, reducing the synthesis of cysteine from homocysteine. Under our experimental conditions, responses to methionine were likely due to a correction of a deficiency of methionine per se rather than of methyl group donors.     

Effects of rumen-protected methionine supplementation and bacterial lipopolysaccharide infusion on nitrogen metabolism and hormonal responses of growing beef steers

Metabolic demand for sulfur-containing AA increases during inflammation in nonruminants. Therefore, Met supplementation may alleviate the negative effects of infection on N balance. Effects of gram-negative bacterial lipopolysaccharide (LPS) and supplemental dietary Met on N balance, serum hormones and haptoglobin, and plasma urea-N and AA were evaluated in 20 Angus-cross steers (BW = 262 +/- 6.3 kg). Treatments (2 x 2 factorial) were infusion of no LPS (-LPS) or a prolonged low dose of LPS (+LPS) and dietary supplementation of no (-MET) or 14 g/d (+MET) of rumen-protected Met (providing 7.9 g/d of dl-Met). Steers were adapted to a roughage-based diet (DMI = 1.4% of BW daily) and supplemental Met for 14 d, and were then infused (1 mL/min via intravenous catheter) with LPS on d 1 (2 microg/kg of BW) and 3 (1 microg/kg of BW) of a 5-d collection period. Blood was collected on d 1, before LPS infusion, and at 2, 4, 6, 8, 10, 12, and 24 h after LPS challenge. Diet samples, feed refusals, feces, and urine were collected daily for 5 d. Rectal temperature and serum concentrations of cortisol, prolactin, tumor necrosis factor-alpha, and haptoglobin increased, whereas thyroxine and triiodothyronine decreased for +LPS vs. -LPS steers (LPS x h; P < 0.01). Plasma urea-N was greater for +LPS than -LPS steers (LPS; P = 0.03), and serum IGF-1 was not affected (P > or = 0.26) by LPS or Met. Plasma concentrations of Thr, Lys, Leu, Ile, Phe, Trp, Asn, Glu, and Orn decreased, plasma Ala increased, and Gly and Ser initially increased, then declined in +LPS vs. -LPS steers (LPS x h; P < or = 0.04). Plasma Met was greater for +MET than -MET steers before LPS infusion, but declined in +MET steers after LPS infusion (LPS x Met x h; P < 0.01). By design, DMI was not different, but DM digested was less (P = 0.04) for +LPS than -LPS steers. Infusion of LPS did not affect (P > or = 0.24) N intake, fecal N excretion, or N digested, but resulted in greater (P < 0.01) urinary N excretion and less (P < 0.01) N retention. The absence of an LPS x Met interaction (P = 0.26) for N retention indicates that supplemental Met does not improve the N utilization of growing beef steers exposed to a gram-negative bacterial endotoxin. Decreases in plasma concentrations of several essential AA in +LPS steers suggest that metabolic demand for these AA likely increased in steers exposed to endotoxin.     

The effect of abomasal methionine supplementation on nitrogen retention of growing steers postruminally infused with casein or nonsulfur-containing amino acids

An experiment was conducted to develop a system useful for measuring methionine requirements of growing steers. Seven ruminally cannulated steers (312 kg, gaining .91 kg/d) were fed a diet based on ammoniated corn cobs, corn starch, molasses and urea. Quantities of N and sulfur-containing amino acids disappearing from the small intestine were 96.0 and 14.8 g/d, respectively. Postruminal infusions of Na-caseinate (CAS) resulted in linear (P less than .01) increases in N retention with values increasing from 30.1 g/d with no postruminal CAS infusion to 39.3, 50.8 and 59.2 g/d (averaged across methionine supplementation) when 100, 200 and 300 g/d CAS were infused. Postruminal infusions of a mixture of crystalline L-amino acids (simulating the nonsulfur-containing essential amino acid pattern of casein; SIM) at levels of 100, 200 and 300 g/d also led to linear increases (P less than .01) in N retention with steers retaining 30.9, 38.9 and 50.5 g N/d (averaged across methionine supplementation), respectively. Postruminal infusion of 12 g/d L-methionine across CAS and SIM infusions improved (P less than .01) N retention by 7.6 g/d but infusion of SIM, which is devoid of sulfur amino acids, also increased N retention. Responses to methionine supplementation was greatest when 200 or 300 g/d SIM were abomasally infused. The data are interpreted to demonstrate that, for steers fed a diet containing little true protein, postruminal supplementation with nonsulfur-containing amino acids tended to increase the ability of growing steers to respond to methionine supplementation.     

Sulfur-containing amino acid requirement of rapidly growing steers

Eight ruminally cannulated steers (294 kg, ADG = 1.3 kg/d) were used in a N retention study (8 x 8 latin-square design) to evaluate sulfur-containing (S) amino acid (AA) requirements for growth. Treatments were abomasal infusions of seven levels of L-methionine (0, 3, 6, 9, 12, 15 and 18 g/d) and one level of DL-methionine (6 g/d). All steers were fed a semipurified diet based on ammoniated corn cobs (DMI = 6.56 kg/d) and were abomasally infused with 400 g/d dextrose and 296.4 g/d of crystalline AA that simulated the non-S-AA pattern of casein. Infusion of 3 g/d supplemental L-methionine maximized N retention in steers. Intestinal flows of absorbable S-AA were determined to be 1.89 g/kg DMI. Breakpoint analysis of retained N as a function of total absorbable S-AA yielded a total S-AA requirement of 14.7 g/d. Nitrogen retention for DL-methionine (36.4 g/d) was not different (P greater than .05) from that for 6 g/d L-methionine (38.8 g/d), but because this value was not in the linear response range, the efficacy of DL-methionine in meeting S-AA needs could not be evaluated. Plasma methionine concentrations increased linearly (P less than .05) in response to L-methionine infusion and were greater (P less than .05) for steers infused with 6 g/d DL-methionine (45.3 microM) than for steers receiving 6 g/d L-methionine (30.5 microM). Plasma cystine increased when up to 9 g/d L-methionine was infused.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of ruminal casein and glucose on forage digestion and urea kinetics in beef cattle

Effects of supplemental glucose and degradable intake protein on nutrient digestion and urea kinetics in steers (Bos taurus) given ad libitum access to prairie hay (4.7% CP) were quantified. Six ruminally and duodenally cannulated steers (initial BW 391 kg) were used in a 4 × 4 Latin square with 2 extra steers. Treatments were arranged as a 2 × 2 factorial and included 0 or 1.2 kg of glucose and 240 or 480 g of casein dosed ruminally once daily. Each period included 9 d for adaptation, 4 d for total fecal and urine collections, and 1 d for ruminal and duodenal sampling. Jugular infusion of (15)N(15)N-urea with measurement of enrichment in urine was used to measure urea kinetics. Glucose reduced forage intake by 18% (P < 0.01), but casein did not affect forage intake (P = 0.69). Glucose depressed (P < 0.01) total tract NDF digestion. Glucose supplementation decreased ruminal pH 2 h after dosing, but the effect was negligible by 6 h (treatment × time; P = 0.01). Providing additional casein increased the ruminal concentration of NH(3), but the increase was less when glucose was supplemented (casein × glucose; P < 0.01). Plasma urea-N was increased (P < 0.01) by additional casein but was reduced (P < 0.01) by glucose. Microbial N flow to the duodenum and retained N increased (P ≤ 0.01) as casein increased, but neither was affected by glucose supplementation. Urea-N entry rate increased (P = 0.03) 50% with increasing casein. Urinary urea-N excretion increased (P < 0.01) as casein increased. The proportion of urea production that was recycled to the gut decreased (P < 0.01) as casein increased. Glucose supplementation decreased (P < 0.01) urinary urea excretion but did not change (P ≥ 0.70) urea production or recycling. The amount of urea-N transferred to the gut and captured by ruminal microbes was less for steers receiving 480 g/d casein with no glucose than for the other 3 treatments (casein × glucose interaction, P = 0.05), which can be attributed to an excess of ruminally available N provided directly to the microbes from the supplement. Overall, the provision of supplemental glucose decreased forage intake and digestibility. Increasing supplemental casein from 240 to 480 g/d increased urea production but decreased the proportion of urea-N recycled to the gut.     

Effects of ruminal and postruminal infusion of starch hydrolysate or glucose on the microbial ecology of the gastrointestinal tract in growing steers

Forty crossbred steers were used to determine the effects of carbohydrate supply site on the indigenous bacteria of the gastrointestinal tract. Steers were fitted with ruminal and abomasal infusion catheters and assigned randomly to one of eight groups in a complete randomized block design. The experimental period was 36 d. Treatments included: 1) a pelleted basal diet fed at 0.163 Mcal ME x (kg BW(0.75)) x 1 x d(-1) (LE); 2) the basal diet fed at 0.215 Mcal ME x (kg BW(0.75)) (-1) x d(-1) (HE); 3) the basal diet fed at 0.163 Mcal ME x (kg BW(0.75))(-1) x d(-1) with ruminal infusion of starch hydrolysate (SH) (RSH); 4) the basal diet fed at 0.163 Mcal ME x (kg BW(0.75))(-1) x d(-1) with abomasal infusion of SH (ASH); and 5) the basal diet fed at 0.163 Mcal ME x (kg BW(0.75))(-1) x d(-1) with abomasal infusion of glucose (AG). The total volume ofinfusate (5 kg x site(-1) x d(-1)) was equalized across treatments and infusion sites by infusion of water. Glucose and SH were infused at rates of 14.35 and 12.64 g x (kg BW(0.75)) x d(-1), respectively. Ruminal, cecal, and fecal samples were obtained on d 36. Ruminal pH was low (5.79) in LE steers and unaffected (P > 0.10) by increased energy intake or carbohydrate infusion. Cecal and fecal pH were 6.93 and 7.00, respectively, for LE steers. Increasing energy intake (P < 0.10) and the rate of carbohydrate infusion (P < 0.01) significantly decreased cecal and fecal pH compared with LE. Ruminal counts of anaerobic bacteria in LE steers were 8.99 log10 cells/g and abomasal carbohydrate infusion had no affect (P > 0.10) on these numbers. However, ASH and AG steers had approximately 1.5 log10 cells/g more (P < 0.01) cecal and fecal anaerobic populations. Ruminal, cecal, and fecal aerobic bacterial counts were 40, 22, and 23%, respectively, lower than anaerobic counts. Generally, aerobic counts responded similarly to the anaerobic counts. Less than 1% of the anaerobic bacteria enumerated in the rumen, cecum, and feces were coliforms, and 97% of the coliforms were Escherichia coli. Carbohydrate infusions resulted in only numerical increases in fecal coliform and E. coli concentrations (P > 0.10). Fecal E. coli were highly acid sensitive in all steers, with less than 1% surviving a 1-h exposure to low pH (2.0). This suggests that cecal or fecal pH is not a good indicator of acid resistance, and it supports the concept that there are other factors that may induce acid resistance.     

Effect of ruminal protein degradability on growth and N metabolism in growing beef steers

The objective of two experiments was to correlate plasma levels of urea N (PUN) and the percentage of urine N in the form of urea (UUN) to weight gain in response to different dietary protein regimens for growing Angus steers. In Exp. 1, 60 steers (302 kg BW) were assigned to various levels of dietary N (control plus supplemental N to provide from 100 to 400 g more crude protein daily) within two sources of supplemental N (soybean meal [SBM] or a mixture of two parts corn gluten meal:one part blood meal [CGM:BM]). In Exp. 2, 27 steers (229 kg BW) were fed two levels of SBM, and half of the steers received growth-promoting implants. Steers were housed in groups of 12 and fed individually for 84 d in both experiments. Corn silage was fed at a restricted rate to minimize orts. Jugular blood and urine samples were collected during the experiments. In Exp. 1, maximal ADG of steers fed SBM (1.0 kg) was reached with 671 g/d total crude protein, or 531 g/d metabolizable protein. Maximal ADG of steers fed CGM:BM (0.91 kg) was reached with 589 g/d total crude protein, or 539 g/d metabolizable protein. The DMI was higher (P < 0.07) for steers fed SBM (6.37 kg/d) than for steers fed CGM:BM (6.14 kg/d). Increasing ruminal escape protein from 36% (SBM) to 65% (CGM:BM) of CP decreased (P < 0.05) endogenous production of urea, as evidenced by lower concentrations of urea in blood and lower UUN. In Exp. 2, increasing supplemental protein from 100 to 200 g/d increased (P < 0.05) ADG and PUN. Implants lowered (P < 0.05) UUN, particularly at the higher level of supplemental protein. Protein supplementation of growing steers can be managed to maintain acceptable ADG yet decrease excretion of urea in the urine.     

Effect of undegradable intake protein supplementation on intake, digestion, microbial efficiency, in situ disappearance, and plasma hormones and metabolites in steers fed low-quality grass hay

Four ruminally and duodenally cannulated beef steers (492 +/- 30 kg) were used in a 4 x 4 Latin square design to evaluate the effect of undegradable intake protein (UIP) supplementation on intake, digestion, microbial efficiency, in situ disappearance, and plasma hormones and metabolites in steers fed low-quality grass hay. The steers were offered chopped (10.2 cm in length) grass hay (6.0% CP) ad libitum and 1 of 4 supplements. Supplemental treatments (1,040 g of DM daily), offered daily at 0800, were control (no supplement) or low, medium, or high levels of UIP (the supplements provided 8.3, 203.8, and 422.2 g of UIP/ d, respectively). The supplements were formulated to provide similar amounts of degradable intake protein (22%) and energy (1.77 Mcal of NE(m)/kg). Blood samples were taken at -2, -0.5, 1, 2, 4, 8, 12, and 24 h after supplementation on d 1 (intensive sampling) and at -0.5 h before supplementation on d 2, 3, 4, and 5 (daily sampling) of each collection period. Contrasts comparing control vs. low, medium, and high; low vs. medium and high; and medium vs. high levels of UIP were conducted. Apparent and true ruminal OM and N digestion increased (P < 0.03) in steers fed supplemental protein compared with controls, but there were no differences (P > 0.26) among supplemental protein treatments. There were no differences (P > 0.11) among treatments for NDF or ADF digestion, or total ruminal VFA or microbial protein synthesis. Ruminal pH was not different (P = 0.32) between control and protein-supplemented treatments; however, ruminal pH was greater (P = 0.02) for supplementation with medium and high compared with low UIP. Daily plasma insulin concentrations were increased (P = 0.004) in protein-supplemented steers compared with controls and were reduced (P = 0.003) in steers fed low UIP compared with steers fed greater levels of UIP. Intensive and daily plasma urea N concentrations were increased (P < 0.01) in protein-supplemented steers compared with controls and increased (P < 0.02) for intensive and daily sampling, respectively, in steers supplemented with medium and high UIP compared with low UIP. Supplemental protein increased apparent and true ruminal OM and N digestion, and medium and high levels of UIP increased ruminal pH compared with the low level. An increasing level of UIP increases urea N and baseline plasma insulin concentrations in steers fed low-quality hay.     

Lysine requirement of growing steers

Eight Limousin-cross steers (355 kg) were used in a replicated 4 x 4 Latin-square designed to estimate lysine requirements. Steers were fed a semipurified diet containing little ruminal escape protein. Treatments were abomasal infusions of 0, 8, 16, or 24 g/day L-lysine. All steers were additionally infused with 400 g/day dextrose and 285.9 g/day of an amino acid mix that contained (g/day) L-methionine (12.0), L-histidine (8.1), L-arginine (10.5), L-threonine (12.0), L-valine (18.0), L-isoleucine (13.8), L-leucine (27.3), L-phenylalanine (28.2), L-glutamic acid (76.5), glycine (76.5) and L-tryptophan (3.0); it had been demonstrated previously that when lysine was included in this infusion mixture, nutritional requirements of steers for maximal N retention were met or exceeded. Nitrogen retention averaged 38 g/day and was not affected by treatment, implying that the lysine requirement of steers was less than the 37.8 g/day lysine estimated to be absorbed from the small intestine when the basal diet was fed.