Limiting amino acids for growing lambs fed a diet low in ruminally undegradable protein

Ruminally cannulated Rambouillet wether lambs were used in three 6 x 6 Latin square experiments (n = 6/experiment) to determine which essential AA limit N retention. Lambs (BW = 36.9 +/- 1.9 kg for Exp. 1, 35.1 +/- 1.4 kg for Exp. 2, and 46.0 +/- 1.3 kg for Exp. 3) were housed in metabolism crates and limit-fed (DMI = approx. 1.8% of BW daily) twice daily a soybean hull-based diet low in ruminally undegradable protein. Treatments for Exp. 1 were continuous abomasal infusions of a solution (500 mL/d) containing 1) no AA (CON), 2) a mixture of 10 essential AA and 2 nonessential AA (10EAA), 3) 10EAA with Met removed, 4) 10EAA with Lys removed, 5) 10EAA with His removed, and 6) 10EAA with Thr removed. Treatments for Exp. 2 were abomasal infusions of 1) CON, 2) 10EAA, 3) 10EAA with Leu, Ile, and Val removed (-BCAA), 4) 10EAA with Arg removed, 5) 10EAA with Phe removed, and 6) 10EAA with Trp removed. Treatments for Exp. 3 were abomasal infusions of 1) CON, 2) 10EAA, 3) -BCAA, 4) 10EAA with Leu removed, 5) 10EAA with Ile removed, and 6) 10EAA with Val removed. All lambs received continuous infusions of acetate and propionate into the rumen and dextrose into the abomasum to supply additional energy. Periods were 7 d: 3 d for adaptation to abomasally infused treatments and 4 d for fecal and urinary collections. Blood samples were collected 3 h after feeding on d 7. In all 3 experiments, N retention was greater (P < 0.10) for lambs receiving 10EAA vs. CON, demonstrating that the basal AA supply from CON was limiting. Removal of each of the essential AA from 10EAA decreased (P < 0.10) their concentrations in plasma (except for Trp), indicating that 10EAA supplied these AA in excess of the animal's requirement. In Exp. 1, N retention (g/d) decreased (P < 0.10) in response to the removal of Met and Thr, but was not affected by removal of Lys and His from 10EAA. In Exp. 2, N retention decreased (P < 0.10) in response to removal of all 3 branched-chain AA, Arg, and Trp, whereas the removal of Phe from 10EAA did not affect N retention. In Exp. 3, N retention decreased (P < 0.10) in response to removal of branched-chain AA and Val, but was not affected by the omission of Leu and Ile from 10EAA. The results of this research demonstrated that Met, Thr, Arg, Trp, and Val limited N retention of lambs fed a diet low in ruminally undegradable protein.     

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.     

Effect of protein source on nitrogen balance and plasma amino acids in exercising horses

Plasma AA in horses fed either an all-hay or a hay and grain diet in a traditional format have not been investigated. Eight horses were divided into 2 groups: a hay group fed only grass hay or a hay and a grain group (HG) fed in a crossover design for two 5-wk periods. After the first period, horses were fasted overnight, followed by feeding with blood sampling every hour for 6 h. A 4-d total fecal and urine collection to evaluate N balance followed. A 10-d washout period separated the 5-wk feeding periods, during which horses switched diets. The second period was also followed by fasting, feeding, blood sampling, and a 4-d collection period. Horses consumed 840 g of CP in the hay group and 865 g of CP in the HG group. Horses in the hay group had a 2.4 ± 2.4 g/d N balance, which was not different from 0 (P = 0.34), whereas horses in the HG group had 5.4 ± 2.4 g/d N balance, which was different from 0 (P = 0.045). Fecal N excretion was greater for the hay group compared with the HG group (hay = 51.1 ± 1.3 g/d and HG = 45.5 ± 1.3 g/d; P = 0.011), and urine N excretion was greater for the HG group compared with the hay group (hay = 79.3 ± 2.8 g/d and HG = 89.2 ± 2.8 g/d; P = 0.026). Plasma AA concentrations were greater in the HG group compared with the hay group for Met (P = 0.001), Lys (P = 0.001), Ile (P = 0.047), Arg (P < 0.001), Gln (P = 0.009), and Orn (P = 0.002). Plasma concentrations were less for the HG group compared with the hay group for Thr (P < 0.001) and Ala (P < 0.001). Plasma concentrations of urea were greater for the HG group compared with the hay group (P < 0.001), whereas 3-methyl-histidine concentrations were greater for the hay group compared with the HG group (P < 0.001). The effect of diet on the excretion of N via feces vs. urine in the hay and HG groups is typical. The early increases in the plasma concentrations of Met, Val, Ile, Leu, Phe, Lys, Arg, and Ala during the postfeeding phase are most likely due to increased foregut digestibility as well as a greater quality AA profile in the grain. The greater concentrations of Thr, Leu, and Val later in the postfeeding phase for the hay group most likely reflects slower digestion because of prolonged consumption time compared with the HG group. Improved N balance observed in the HG group supports the fact that the HG group had more available AA via the AA profile and foregut digestibility of the HG diet. Despite the fact that both groups consumed similar amounts of CP, the AA profile and availability affected N balance.     

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.     

Muscle growth and plasma concentrations of amino acids, insulin-like growth factor-I, and insulin in growing pigs fed reduced-protein diets

Twenty barrows were used to determine if partial replacement of protein-bound AA with crystalline AA (CAA) reduces AA use for muscle tissue and whole-body growth. Barrows (44.2 +/- 1.3 kg of BW) were assigned to 4 diets in a randomized complete block design. Diets consisted of 16.1% CP with no CAA, and 12.8, 10.1, and 7.8% CP containing CAA. As the CP concentration decreased, CAA were gradually increased to meet requirements on a true ileal digestibility basis. Barrows were weighed on d 0 and 13. Blood samples were collected before the morning feeding on d 0, 6, and 12 (prefeeding), and 2 h after the morning feeding on d 13 (postfeeding). Pigs were euthanized on d 13, and liver and right LM were removed and weighed. The reduction in the dietary CP concentration linearly decreased (P < 0.01) ADG, G:F, LM weight, and the CP content of LM. Reducing the CP concentration decreased pre- and postfeeding plasma concentrations of IGF-I (linear, P < 0.01) and insulin (linear, P < 0.10). The reduction in the dietary CP concentration increased prefeeding plasma concentrations of Ala, Gln, Gly, and total AA but decreased Arg, Asn, His, Ile, Phe, Trp, and Tyr (linear, P < 0.05). Plasma concentration of total indispensable AA decreased initially and increased thereafter as the dietary CP concentration decreased from 16.1 to 7.8% (quadratic, P < 0.01). The reduction in the dietary CP concentration increased postfeeding plasma concentrations of Ala, Lys, Met (linear, P < 0.01), and Gly (linear, P = 0.073) and decreased Asn, Ser, Tyr, Arg, His, and Leu (linear, P < 0.05). Plasma concentrations of Ile, Phe, Thr, Trp, and Val decreased initially and increased thereafter as the dietary CP concentration decreased from 16.1 to 7.8% (quadratic, P < 0.05). In muscle tissue, concentrations of free Ala, Asp, Glu, Gln, Gly, and Lys increased (linear, P < 0.05) as the dietary CP concentration decreased. Concentrations of free His, Ile, Phe, Thr, Trp, and Val in muscle tissue decreased initially and increased thereafter as the dietary CP concentration decreased from 16.1 to 7.8% (quadratic, P < 0.05). In summary, the reduction in the dietary protein-bound AA decreased whole-body and LM growth, altered the free AA pool profile in muscle tissue, and decreased plasma insulin and IGF-I. As the replacement of protein-bound AA with CAA increased, 1) free Ala and Gln in muscle tissue increased, indicating an increase of muscle tissue protein breakdown; and 2) utilization of indispensable AA in muscle tissue decreased.     

Effects of energy supply on leucine utilization by growing steers at two body weights

The effects of energy supplementation on Leu utilization in growing steers were evaluated in 2 experiments by using 6 ruminally cannulated Holstein steers. In Exp. 1, steers (initial BW = 150 +/- 7 kg) were limit-fed (2.3 kg of DM/d) a diet based on soybean hulls and received a basal ruminal infusion of 100 g of acetate/d, 75 g of propionate/d, and 75 g of butyrate/d, as well as abomasal infusions of 200 g of glucose/d and a mixture (215 g/d) containing all essential AA except Leu. Treatments were arranged as a 3 x 2 factorial, with 3 amounts of Leu infused abomasally (0, 4, and 8 g/d) and supplementation of diets with 2 amounts of energy (0 and 1.9 Mcal/d of GE). Supplemental energy was supplied by ruminal infusion of 100 g of acetate/ d, 75 g of propionate/d, and 75 g of butyrate/d, as well as abomasal infusion of 200 g of glucose/d to provide energy to the animal without affecting the microbial protein supply. When no supplemental energy was provided, Leu supplementation increased N balance, with no difference between 4 and 8 g/d of Leu (24.5, 27.0, and 27.3 g/d for 0, 4, and 8 g/d of Leu), but when additional energy was supplied, N retention increased linearly in response to Leu (25.6, 28.5, and 31.6 g/d for 0, 4, and 8 g/d of Leu; Leu x energy interaction, P = 0.06). The changes in N balance were the result of changes in urinary N excretion. The greater Leu retentions in response to energy supplementation when Leu was the most limiting nutrient indicate that energy supplementation improved the true efficiency of Leu utilization. In addition, supplemental energy increased the gross efficiency of Leu utilization when the Leu supply was not limiting by increasing the maximal rates of protein deposition. Experiment 2 was similar to Exp. 1, but steers had an initial BW of 275 +/- 12 kg and were limit-fed at 3.6 kg of DM/d. Retention of N was not affected (P = 0.22) by Leu supplementation, indicating that Leu did not limit protein deposition. Energy supply increased N retention (P < 0.01) independently of Leu supplementation (33.0 vs. 27.8 g/d). Overall, energy supplementation improved Leu utilization by modestly increasing N retention when Leu was limiting and by increasing the ability of steers to respond to the greatest amount of supplemental Leu. We conclude from these results that the assumption of a constant efficiency of AA utilization is unlikely to be appropriate for growing steers.     

Isoleucine requirement of 80- to 120-kilogram barrows fed corn-soybean meal or corn-blood cell diets

Six experiments were conducted to validate an Ile-deficient diet and determine the Ile requirement of 80- to 120-kg barrows. Experiment 1 had five replications, and Exp. 2 through 6 had four replications per treatment; all pen replicates had four crossbred barrows each (initial BW were 93, 83, 85, 81, 81, and 88 kg, respectively). All dietary additions were on an as-fed basis. In Exp. 1, pigs were fed a corn-soybean meal diet (C-SBM) or a corn-5% blood cell (BC) diet with or without 0.26% supplemental Ile (C-BC or C-BC+Ile) in a 28-d growth assay. On d 14, pigs receiving the C-BC diet were taken off experiment as a result of a severe decrease in ADFI. Growth performance did not differ for pigs fed C-SBM or C-BC + Ile (P = 0.36) over the 28-d experiment. In Exp. 2, pigs were fed the C-BC diet containing 0.24, 0.26, 0.28, 0.30, or 0.32% true ileal digestible (TD) Ile for 7 d in an attempt to estimate the Ile requirement using plasma urea N (PUN) as the response variable. Because of incremental increases in ADFI as TD Ile increased, PUN could not be used to estimate the Ile requirement. In Exp. 3, pigs were fed the C-BC diet containing 0.28, 0.30, 0.32, 0.34, or 0.36% TD Ile. Daily gain, ADFI, and G:F increased linearly (P < 0.01) as Ile increased in the diet. Even though there were no effects of TD Ile concentration on 10th rib fat depth or LM area, kilograms of lean increased linearly (P < 0.01) as TD Ile level increased. In Exp. 4, pigs were fed a C-SBM diet containing 0.26, 0.31, or 0.36% TD Ile. There were no differences in ADFI or ADG; however, G:F increased linearly (P = 0.02), with the response primarily attributable to the 0.31% Ile diet. In Exp. 5, pigs were fed 0.24, 0.27, 0.30, 0.33, or 0.36% TD Ile in a C-SBM diet. There were no differences in growth performance; however, average backfat, total fat, and percentage of fat increased quadratically (P < 0.10) with the addition of Ile. In Exp. 6, pigs were fed a 0.26% TD Ile C-SBM diet with or without crystalline Leu and Val to simulate the branched-chain AA balance of a C-BC diet. There were no differences in ADFI or ADG, but G:F increased (P = 0.09) when Leu and Val were added. In summary, the Ile deficiency of a C-BC diet can be corrected by the addition of Ile, and because ADFI was affected by Ile addition, the PUN method was not suitable for assessing the Ile requirement. The TD Ile requirement for 80- to 120-kg barrows for maximizing growth performance and kilograms of lean is not < 0.34% in a C-BC diet, but may be as low as 0.24% in a C-SBM diet.     

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.     

Effect of dietary excess of branched‐chain amino acids on performance and serum concentrations of amino acids in growing pigs

Depressed performance and availability of some amino acids (AA) in pigs fed excess Leu diets appear to be related to lower feed intake. Surplus Ile and Val may help to overcome this effect. An experiment was conducted with 24 pigs (31.8 ± 1.2 kg initial BW) to evaluate the effect of dietary excess of either Leu alone or with surplus Ile and Val on performance and serum concentration (SC) of essential AA. Treatments were as follows: T1, basal diet; T2, basal plus 0.43% L‐Leu (excess Leu); T3, basal added with 0.43% L‐Leu, plus 0.20% L‐Ile and 0.25% L‐Val (excess LIV). The basal diet was formulated to contain 0.90% standardized ileal digestible Lys and added with crystalline L‐Lys, L‐Thr, DL‐Met, L‐Trp, L‐Leu, L‐Ile, L‐His and L‐Val to create essential AA:Lys ratios close to an ideal protein for growing pigs. All pigs were fed the same amount of feed twice a day (average, 3.42× the requirement of NEm). Blood samples were collected at 2.5 (absorptive) and 11.0 h (post‐absorptive) post‐prandial to analyse SC of AA. Excess of either Leu or LIV did not affect growth rate nor feed conversion. Excess Leu increased Leu SC and decreased Ile and Val SC (p < 0.05) at both absorptive and post‐absorptive phases, but excess LIV restored the SC of Ile and Val. The SC of other essential AA was not affected by excess of either Leu or LIV. The SC of all AA during absorptive, on average, was about two times higher than that of post‐absorptive phase. These results suggest that the reduced availability (SC) of Ile and Val in pigs consuming excess Leu diets is attributed to a reduced absorption and increased cellular degradation rates of them.     

Regulation of taste-active components of meat by dietary branched-chain amino acids; effects of branched-chain amino acid antagonism

1. The effects of dietary branched-chain amino acids (BCAAs) including leucine (Leu), isoleucine (Ile) and valine (Val) on taste-active components, especially free glutamate (Glu), in meat were investigated. 2. Broiler chickens (28 d old) were given varied dietary BCAA levels for 10 d before marketing. Dietary BCAA content ratios were either 100:100:100 (Low Leu group), 150:100:100 (Control group) or 150:150:150 (High Ile + Val group) for Leu:Ile:Val (% of each BCAA requirement according to NRC, 1994). Taste-related components of meat (free amino acids and ATP metabolites) and sensory scores of meat soup were estimated. 3. Free Glu content, the main taste-active component of meat, was significantly increased by dietary BCAA. Compared to the Control group, free Glu content increased by 30% in the High Ile + Val group. However, the inosine monophosphate (IMP) content in meat did not change among groups. 4. Sensory evaluation of meat soups showed that Control and High Ile + Val groups had different meat flavours. The sensory score of overall taste intensity was significantly higher in the High Ile + Val group. 5. These results suggest that dietary BCAA concentrations regulate free Glu in meat. Increasing dietary Ile + Val induces an increase in free Glu content of meat, improves meat taste and is more effective for increasing free Glu content in meat than decreasing dietary Leu level.     

Biochemical and morphological developments are partially impaired in intestinal mucosa from growing pigs fed reduced-protein diets supplemented with crystalline amino acids

The objective of this study was to determine if a reduction in dietary CP, with partial replacement of the intact protein with crystalline AA (CAA), would alter growth, morphology, and free or peptide-bound AA concentrations of intestinal mucosa in growing pigs. Twenty-four barrows (37.0 +/- 1.5 kg of BW) were fed 1 of 4 diets for 24 d: 16.1% CP with no CAA, or 12.8, 10.1, or 7.8% CP (analyzed values, as-fed) containing CAA. As CP decreased, CAA were gradually increased to meet requirements on a true ileal digestible basis. Pigs were euthanized 2 h postmeal on d 24, and mucosal samples from duodenum, jejunum, and ileum were collected. Reducing dietary CP decreased ADG, G:F, and final weight (linear, P < 0.05). With reduced dietary CP, mucosal protein concentration decreased in the jejunum (quadratic, P < 0.05) and tended to decrease in the ileum (linear, P = 0.062). Reduction of the dietary CP concentration from 16.1 to 7.8% tended to decrease the crypt depth (linear, P < 0.10) and decreased villus width (linear, P < 0.05) in duodenum and jejunum mucosa but did not reduce villus height or villus surface area in any regions of the small intestine. In the duodenum, a reduction in dietary CP increased free Lys, Met, and Thr (linear, P < 0.05) and peptide-bound Lys and Thr (quadratic, P < 0.10). In the jejunum, reducing CP decreased free Cys (linear P < 0.05) and tended to decrease free Asn and His (linear, P < 0.10) and peptide-bound His (quadratic, P = 0.061) and Ile, Leu, and Val (linear, P < 0.10). In the ileum, reducing CP decreased free Asn, Ser, Tyr, Arg, His, Phe (linear, P < 0.05), and Leu (linear, P = 0.054) and peptide-bound Gly and Ser (linear, P < 0.05) and tended to decrease peptide-bound Ile, Leu, Phe, Val (linear, P < 0.10), and Lys (linear P < 0.05). In conclusion, reduced-CP diets supplemented with CAA lead to a reduction in growth performance, associated with biochemical and morphological modifications of the intestinal mucosa.     

Isoleucine and valine supplementation of a low-protein corn-wheat-soybean meal-based diet for piglets: growth performance and nitrogen balance

The effects of Ile and Val supplementation of a low-CP, corn-wheat-soybean meal-based piglet diet on growth performance, incidence of diarrhea, and N balance were studied using 60 Landrace x Duroc male piglets in a 4-wk experiment. The 60 individually caged piglets were divided into 5 dietary treatments, each consisting of 12 piglets. Diet 1 was a positive control diet (20% CP); diet 2 was a low-CP negative control diet (17% CP); diets 3, 4, and 5 were low-CP diets to which Ile, Val, or the combination of Ile and Val were added, respectively. All diets were supplemented with Lys, Met, Thr, and Trp to provide the required concentrations of these AA according to the 1998 NRC. Average daily gain and ADFI were similar among pigs fed the positive control, Val-added, and the Val plus Ile-added diets. On wk-2 and wk-4, fecal score was greater (softer feces) in piglets fed the 20% CP level compared with the remaining treatments (P < 0.01). Nitrogen intake was decreased (P < 0.0001) in pigs fed diets containing low levels of CP compared with pigs fed the 20% CP diet. Fecal N excretion (g/d) was decreased (P < 0.05) in piglets fed low-CP diets at wk 1 and wk 4 of feeding, and in urine at wk 4 of feeding. Crude protein levels or AA supplementation had no effect on N retention efficiencies. These results indicate that the supplementation of Val alone, or in combination with Ile, to a low-CP piglet diet with adequate levels of Lys, Met, Thr, and Trp is necessary to achieve maximum performance in pigs consuming corn-wheat-soybean meal-based diets.     

Effects of alfalfa extract, anise, capsicum, and a mixture of cinnamaldehyde and eugenol on ruminal fermentation and protein degradation in beef heifers fed a high-concentrate diet

Four Holstein heifers (360 +/- 22 and 450 +/- 28 kg of BW in Exp. 1 and 2, respectively) fitted with ruminal trocars were used in 4 x 4 Latin square designs to evaluate the effects on ruminal microbial fermentation of the following: Exp. 1, no additive, alfalfa extract (30 g/d, AEX), a mixture of cinnamaldehyde (0.18 g/d) and eugenol (0.09 g/d; CIE1), and AEX and CIE1 in combination; and Exp. 2, no additive, anise oil (2 g/d), capsicum oil (1 g/d), and a mixture of cinnamaldehyde (0.6 g/d) and eugenol (0.3 g/d). Heifers were fed a 90:10 concentrate:barley straw diet (16% CP; 25% NDF) for ad libitum intake. Each period consisted of 15 d for adaptation and 6 d for sampling. On d 16 to 18, DM and water intakes were measured. On d 19 to 21 ruminal contents were sampled at 0, 3, 6, 9, and 12 h after feeding to determine ruminal pH and the concentrations of VFA, L-lactate, large peptides, small peptides plus AA (SPep+AA), and ammonia N. On d 20 and 21, samples of ruminal fluid were collected at 0 and 3 h after feeding to determine protozoal counts. In Exp. 1, CIE1 and AEX decreased (P < 0.05) total DMI, concentrate DMI, and water intake. The increase (P < 0.05) in SPep+AA and the decrease (P < 0.05) in ammonia N when supplementing CIE1 suggest that deamination was inhibited. Treatment AEX increased (P < 0.05) the acetate to propionate ratio, which is less efficient for beef production. Treatment CIE1 increased (P < 0.05) counts of holotrichs. Effects of AEX and CIE1 were not additive for many of the measured metabolites. In Exp. 2, treatments had no effect on ruminal pH, total VFA concentration, and butyrate proportion. The capsicum oil treatment increased (P < 0.05) DMI, water intake, and SPep+AA N concentration and decreased (P < 0.05) acetate proportion, branched-chain VFA concentration, and large peptide N concentration. The cinnamaldehyde (0.6 g/d) and eugenol (0.3 g/d) treatment decreased (P < 0.05) water intake, acetate proportion, branched-chain VFA, L-lactate, and ammonia N concentrations and increased (P < 0.05) propionate proportion and SPep+AA N concentration. The anise oil treatment decreased (P < 0.05) acetate to propionate ratio, branched-chain VFA and ammonia N concentrations, and protozoal counts. The results indicate that at the doses used a mixture of cinnamaldehyde and eugenol, anise oil, and capsicum oil may be useful as modifiers of rumen fermentation in beef production systems.     

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.     

Baggs ewes adapt to maternal undernutrition and maintain conceptus growth by maintaining fetal plasma concentrations of amino acids

Adequate delivery of AA is essential for normal fetal growth and development. Recently, we reported that when ewes from the University of Wyoming flock (farm flock with adequate nutrition) were fed 50% (nutrient-restricted) or 100% (control-fed) of the NRC-recommended nutrient requirements between d 28 and 78 of gestation, fetal weights as well as concentrations of most AA in maternal and fetal blood were substantially reduced in nutrient-restricted vs. control-fed pregnancies. The current study utilized Baggs ewes, which were selected under a markedly different production system (range flock with limited nutrition), to test the hypothesis that adaptation of ewes to nutritional and environmental changes may alter placental efficiency and conceptus nutrient availability in the face of maternal nutrient restriction. Baggs ewes received 50 or 100% of the NRC nutrient requirements between d 28 and 78 of pregnancy. On d 78, maternal uterine arterial and fetal umbilical venous blood samples were obtained, and the ewes were euthanized. Amino acids and their metabolites (ammonia, urea, and polyamines) in plasma were analyzed using enzymatic and HPLC methods. The results showed that maternal plasma concentrations of 9 AA (Asp, Ile, Leu, Lys, Orn, Phe, Thr, Trp, and Val) as well as maternal and fetal plasma concentrations of ammonia and urea were reduced (P < 0.05) in nutrient-restricted compared with control-fed Baggs ewes. However, fetal plasma concentrations of all AA and polyamines did not differ (P = 0.842) between the 2 groups of ewes. Collectively, these findings suggest that Baggs ewes, by adapting to the harsh conditions and limited nutrition under which they were selected, were able to maintain fetal concentrations of AA in the face of a maternal nutrient restriction through augmenting placental efficiency.     

Improving performance of finishing pigs with added valine,isoleucine, and tryptophan: validating a meta-analysis model

Based on results of a recent meta-analysis, we hypothesized that increased dietary Val, Ile, or Trp could correct possible amino acid interactions because of excess Leu in diets containing high levels of corn protein, namely dried distiller’s grains with solubles (DDGS). A total of 1,200 pigs (PIC TR4 × (Fast LW × PIC L02); initially 33.6 ± 0.6 kg) were used in a 103-d study. The 6 dietary treatments were corn–soybean meal (SBM)-DDGS-based as follows: (1) high SBM and low level of l-Lys HCl (HSBM), (2) high l-Lys HCl and moderate Ile, Val, Trp (AA above NRC 2012 estimates; NC), (3) moderate l-Lys HCl and high Ile, Val, and Trp (PC), and PC with either increased (4) L-Val (PC+Val), (5) L-Ile (PC+Ile), or (6) L-Trp (PC+Trp). Pigs fed the NC diet were predicted to have the poorest average daily gain (ADG), the PC diet to be intermediate, and pigs fed the HSBM, PC+Val, PC+Ile, and PC+Trp have the same and highest predicted ADG. In the grower period (34 to 90 kg), ADG was greater (Ρ < 0.05) for the pigs fed HSBM and PC+Val diets than the NC with pigs fed other diets intermediate. Pigs fed HSBM were more (Ρ < 0.05) efficient (G:F) than the NC and PC with pigs fed other diets intermediate. In the finisher period (90 to 136 kg), ADG was greater (Ρ < 0.05) for pigs fed PC+Ile than that of the NC with pigs fed other diets intermediate. Pigs fed PC+Val had greater (Ρ < 0.05) average daily feed intake (ADFI) than the NC with pigs fed other diets intermediate. However, PC+Ile pigs were more (Ρ < 0.05) efficient than PC+Val with pigs fed other diets intermediate. Overall, ADG was greater (Ρ < 0.05) for pigs fed HSBM, PC+Val, and PC+Ile diets than the NC with pigs fed other diets intermediate. Pigs fed the PC+Val diet had greater (Ρ < 0.05) ADFI than the NC with pigs fed other diets intermediate. No differences were detected between treatments for overall G:F or other carcass characteristics. In conclusion, increasing Val or Ile in high l-Lys-HCl-DDGS-based diets improved growth performance compared with pigs fed diets containing high levels of l-Lys HCl without added Val and Ile. These results present evidence that the recently developed meta-analysis can predict the relative differences in overall ADG for pigs fed the NC, PC, PC+Val, and PC+Ile diets; however, the predicted G:F was less accurate. The data demonstrate that the negative effects of high Leu concentrations in corn-DDGS-based diets can be reversed by increasing the ratios of Val and Ile relative to Lys.     

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.     

Supplemental methionine and urea for gestating beef cows consuming low quality forage diets

A study was conducted to evaluate Met requirements of late-gestation beef cows consuming low quality forages on the premise that inadequate supply of metabolizable AA may limit protein accretion during pregnancy. Five ruminally cannulated, multiparous late-gestation beef cows (490 +/- 27 kg), of predominantly Angus (> or =75%) with Hereford and Simmental breeding, were used in a 5 x 5 Latin square experiment to evaluate the effects of postruminal dl-Met supplementation on N retention, serum metabolites, and plasma AA concentrations during the third trimester of pregnancy. The basal diet was fed individually, and weights of refusals were recorded for N intake determination. Treatments consisted of no urea, urea (0.053 +/- 0.002 g/kg of BW daily), urea + 5 g of Met/d, urea + 10 g of Met/d, and urea + 15 g of Met/d. Cows were adapted to the experimental diet 30 d before the beginning of the study, with periods lasting for 14 d; 4 d to allow for clearance of the previous treatment effects, 4 d for adaptation to the treatments, and 6 d for total fecal and urine collection. Blood samples were collected every 4 h on d 13 of each period for analysis of serum metabolites and plasma AA. Inclusion of urea increased DM and OM intakes (urea vs. no urea; P = 0.05), but no further improvement in intake was observed with inclusion of Met. Serum urea concentrations increased with inclusion of urea (P = 0.03) and responded quadratically (P = 0.06) when Met was added, with the lowest concentration observed in the urea + 5 g of Met/d treatment. More N was retained with the inclusion of urea (P = 0.04), and N retention increased linearly (P = 0.07) with inclusion of Met. Plasma Met concentration increased linearly (P < 0.01) with inclusion of Met. These data suggest that Met was a limiting AA and that supplementation of a combination of urea and 5 g/d of rumen-protected Met to low quality, forage diets will improve N retention and promote protein accretion during late pregnancy.     

Valine and soleucine requirement of 20- to 45-kilogram pigs

Three experiments were conducted to determine the Val and Ile requirements in low-CP, corn-soybean meal (C-SBM) AA-supplemented diets for 20- to 45-kg pigs. All experiments were conducted for 26 to 27 d with purebred or crossbred barrows and gilts, which were blocked by initial BW. Treatments were replicated with 5 or 6 pens of 3 or 4 pigs per pen. At the beginning of Exp. 1 and the end of all experiments, blood samples were obtained from all pigs to determine plasma urea N (PUN) concentrations. All diets were C-SBM with 0.335% supplemental Lys to achieve 0.83% standardized ileal digestible (SID) Lys, which is the Lys requirement of these pigs. In Exp. 1, 0, 0.02, 0.04, 0.06, 0.08, or 0.10% L-Val was supplemented to achieve 0.51, 0.53, 0.55, 0.57, 0.59, or 0.61% dietary SID Val, and Thr, Trp, Met, and Ile were supplemented to maintain Thr:Lys, Trp:Lys, TSAA:Lys, and Ile:Lys ratios of 0.71, 0.20, 0.62, and 0.60, respectively. Also, supplemental Gly and Glu were added to all diets to achieve 1.66% Gly + Ser and 3.28% Glu, which is equal to the Gly + Ser and Glu content of a previously validated positive control diet that contained no supplemental AA. Treatment differences were considered significant at P < 0.10. Valine addition increased ADG, ADFI, and G:F in pigs fed 0.51 to 0.59% SID Val (linear, P < 0.08), but ADG and ADFI were decreased at 0.61% SID Val (quadratic, P ≤ 0.10). On the basis of ADG and G:F, the SID Val requirement is between 0.56 and 0.58% in a C-SBM diet supplemented with AA. In Exp. 2 and 3, 0, 0.02, 0.04, 0.06, or 0.08% L-Ile was supplemented to achieve 0.43, 0.45, 0.47, 0.49, or 0.51% dietary SID Ile, and Thr, Trp, Met, and Ile were supplemented to maintain Thr:Lys, Trp:Lys, TSAA:Lys, and Val:Lys ratios of 0.71, 0.20, 0.62, and 0.74, respectively. Also, supplemental Gly and Glu were added to achieve 1.66% Gly + Ser and 3.28% Glu as in Exp. 1. Data from Exp. 2 and 3 were combined and analyzed as 1 data set. Daily BW gain, ADFI, and G:F were not affected by Ile additions to the diet; however, ADFI was decreased among pigs fed the diet with 0.45% SID Ile (P < 0.10) compared with pigs fed the 0.43% SID Ile diet. Broken-line analysis requirements could not be estimated for the combined data from Exp. 2 or 3. The results of this research indicate that the SID Val requirement is between 0.56 to 0.58% (0.67 to 0.70 SID Val:Lys), and the Ile requirement is adequate at 0.43% SID Ile (0.52 SID Ile:Lys) for 20- to 45-kg pigs.