We evaluated the effect of increasing amounts of rumen-degradable intake protein (DIP) on urea kinetics in steers consuming prairie hay. Ruminally and duodenally fistulated steers (278 kg of BW) were used in a 4 × 4 Latin square and provided ad libitum access to low-quality prairie hay (4.9% CP). The DIP was provided as casein dosed ruminally once daily in amounts of 0, 59, 118, and 177 mg of N/kg of BW daily. Periods were 13 d long, with 7 d for adaptation and 6 d for collection. Steers were in metabolism crates for total collection of urine and feces. Jugular infusion of ¹⁵N¹⁵N-urea, followed by determination of urinary enrichment of ¹⁵N¹⁵N-urea and ¹⁴N¹⁵N-urea was used to determine urea kinetics. Forage and N intake increased (linear, P < 0.001) with increasing DIP. Retention of N was negative (−2.7 g/d) for steers receiving no DIP and increased linearly (P < 0.001; 11.7, 23.0, and 35.2 g/d for 59, 118, and 177 mg of N/kg of BW daily) with DIP. Urea synthesis was 19.9, 24.8, 42.9, and 50.9 g of urea-N/d for 0, 59, 118, and 177 mg of N/kg of BW daily (linear, P = 0.004). Entry of urea into the gut was 98.9, 98.8, 98.6, and 95.9% of production for 0, 59, 118, and 177 mg of N/kg of BW daily, respectively (quadratic, P = 0.003). The amount of urea-N entering the gastrointestinal tract was greatest for 177 mg of N/kg of BW daily (48.6 g of urea-N/d) and decreased (linear, P = 0.005) to 42.4, 24.5, and 19.8 g of urea-N/d for 118, 59, and 0 mg of N/kg of BW daily. Microbial incorporation of recycled urea-N increased linearly (P = 0.02) from 12.3 g of N/d for 0 mg of N/kg of BW daily to 28.9 g of N/d for 177 mg of N/kg of BW daily. Provision of DIP produced the desired and previously observed increase in forage intake while also increasing N retention. The large percentage of urea synthesis that was recycled to the gut (95.9% even when steers received the greatest amount of DIP) points to the remarkable ability of cattle to conserve N when fed a low-protein diet. Copyright © 2008. . Copyright 2008 Journal of Animal Science

In vitro studies and a lactation trial were conducted to investigate the effects of fibrolytic enzyme mixtures at different inclusion amounts. Seven enzymes in amounts designed to mimic addition of 1, 5, 15, or 30 g/d to dairy diets were incubated in vitro with either soybean hulls or alfalfa for 24 or 48 h. Enzyme treatments generally increased in vitro dry matter disappearance (IVDMD), but not volatile fatty acid production. For some enzyme mixtures, lesser amounts of enzymes led to greater increases in IVDMD, whereas for others there were no differences among the amounts tested. The enzyme mixture with the most cellulase activity was the most effective enzyme in improving IVDMD. In additional in vitro experiments, the same enzymes were used at an amount of 5 g/d, as well as at other amounts that showed promising responses in the first trial. Preincubation of substrates with enzymes before fermentation also was tested. Alfalfa, soybean hulls, corn silage, and corn gluten feed were used as substrates. Preincubation of the substrate with enzymes for 18 h before in vitro fermentation improved IVDMD. The effect on substrate solubilization of incubating substrates with the enzymes but without rumen fluid was also studied. Addition of enzymes to substrates without subsequent fermentation did not solubilize significant amounts of dry matter, indicating that the positive effect of preincubation cannot be attributed directly to hydrolysis of substrates before the in vitro fermentation with ruminal microbes. The fibrolytic enzyme that appeared most promising in vitro did not affect lactational performance when fed to dairy cows.

We evaluated the effect of frequency and amount of rumen-degradable intake protein (DIP) on urea kinetics in steers consuming prairie hay. Five ruminally and duodenally fistulated steers (366 kg of BW) were used in a 5 × 5 Latin square and provided ad libitum access to low-quality prairie hay (4.7% CP). Casein was provided daily in amounts of 61 and 183 mg of N/kg of BW (61/d and 183/d) and every third day in amounts of 61, 183, and 549 mg of N/kg of BW per supplementation event (61/3d, 183/3d, and 549/3d). Periods were 18-d long with 9 d for adaptation and 9 d for collection. Steers were in metabolism crates for total collection of urine and feces. Jugular infusion of ¹⁵N¹⁵N-urea followed by determination of urinary enrichment of ¹⁵N¹⁵N-urea and ¹⁴N¹⁵N-urea was used to determine urea kinetics. Treatment means were separated to evaluate the effects of increasing DIP supplementation and the effects of frequency at the low (61/d vs. 183/3d) and at the high (183/d vs. 549/3d) amounts of DIP provision. Forage OM and total digestible OM intakes were linearly (P ≤ 0.05) increased by increasing DIP provision but were not affected by frequency of supplementation at either the low or high amounts. Production and gut entry of urea linearly (P ≤ 0.006) increased with DIP provision and tended to be greater (P ≤ 0.07) for 549/3d than 183/d but were not different between 61/d and 183/3d. Microbial N flow to the duodenum was linearly (P < 0.001) increased by increasing DIP provision. Additionally, 183/d resulted in greater (P = 0.05) microbial N flow than 549/3d. Incorporation of recycled urea-N into microbial N linearly (P = 0.04) increased with increasing DIP. Microbial incorporation of recycled urea-N was greater for 549/3d than 183/d, with 42 and 23% of microbial N coming from recycled urea-N, respectively. In contrast, there was no difference due to frequency in the incorporation of recycled urea-N by ruminal microbes at the low level of supplementation (i.e., 61/d vs. 183/3d). This study demonstrates that urea recycling plays a substantial role in the N supply to the rumen and to the animal, particularly in steers supplemented infrequently with high levels of protein. Copyright © 2008. . Copyright 2008 Journal of Animal Science

An experiment was conducted to determine the relationship between feeding ractopamine and different amounts of MP on growth and carcass characteristics of feedlot heifers. Seventy-two crossbred heifers (475 kg of initial BW) were fed individually a diet based on steam-flaked corn for ad libitum intake for 29 d. Heifers were implanted with 140 mg of trenbolone acetate and 14 mg of estradiol-17β 60 d before the experiment. Treatments were arranged as a 2 × 3 factorial and included 0 or 200 mg of ractopamine-HCl (23 ppm)/ d, and urea, solvent soybean meal, or expeller soybean meal (ESBM) as the predominant protein supplement. The amounts of MP supplied by the urea, solvent soybean meal, and ESBM diets were 688, 761, and 808 g/ d, respectively, calculated according to level 1 of the NRC model. Body weights were obtained 1 d before ractopamine feeding and at slaughter. Blood samples were obtained 1 d before starting the experiment and 13 d later. Ractopamine improved ADG, efficiency of gain, carcass-adjusted ADG, and carcass-adjusted efficiency of gain (P < 0.01). For ADG, heifers demonstrated a ractopamine × protein source interaction (P < 0.05); heifers not fed ractopamine had greater ADG when fed ESBM than when fed urea, whereas for heifers fed ractopamine there were no differences (P ≥ 0.10) among protein supplements. This interaction was not observed for carcass-adjusted ADG (P = 0.60). Final live weights (P = 0.02) and carcass weights (P = 0.01) were greater with ractopamine feeding. Carcass marbling scores and yield grades were not affected by ractopamine or protein source (P ≥ 0.39). Plasma total α-amino N and glucose concentrations decreased more from pretreatment concentrations when heifers were fed ractopamine (P < 0.05). Feeding ractopamine to heifers for 28 d before slaughter improved ADG and efficiency of gain without any large effects on carcass characteristics. The MP supply does not need to be increased from that provided by finishing diets based on steam-flaked corn with urea as the primary N supplement to allow the maximal response to ractopamine by finishing heifers. Copyright © 2006. . Copyright 2006 Journal of Animal Science

Responses to pharmacological doses of niacin, an agonist for GPR109A (niacin receptor), were different in cattle than in humans and rodents. Thus, the tissue distribution of GPR109A was investigated in cattle. Samples of tail head fat, back fat, perirenal fat, longissimus muscle, and liver were analyzed for abundance of GPR109A mRNA by quantitative real-time reverse transcription-PCR and for abundance of GPR109A protein by Western blotting. Niacin receptor transcript and protein were detected in all tissues analyzed. The mRNA for GPR109A was more abundant in liver than in the other tissues sampled (GPR109A:RPS9 mRNA abundance = 0.56 in liver compared with 0.06 in longissimus muscle, 0.15 in kidney fat, 0.11 in back fat, 0.23 in tail head fat; standard error of the mean = 0.028). Additionally, mRNA for GPR109A was found (GPR109A:RPS9 mRNA abundance ≥ 0.004) in each of the 5 regions of bovine brain that were analyzed: cerebral cortex, cerebellum, thalamus, hypothalamus, and brain stem. Evaluation of liver tissue by immunofluorescence suggested that GPR109A was expressed in parenchymal cells and not localized exclusively to immune-system cells. Finally, analysis of the putative bovine GPR109A sequence verified that AA residues required for binding niacin in human GPR109A are conserved, suggesting that the bovine sequence identified encodes a functional niacin receptor. The identification of GPR109A in bovine liver, muscle, and brain is a novel finding.

Creatine stores high-energy phosphate bonds in muscle, which is critical for muscle activity. In animals, creatine is synthesized in the liver from guanidinoacetic acid (GAA) with methylation by S-adenosylmethionine. Because methyl groups are used for the conversion of GAA to creatine, methyl group deficiency may occur as a result of GAA supplementation. With this study, the metabolic responses of cattle to post-ruminal supplementation of GAA were evaluated with and without methionine (Met) supplementation as a source of methyl groups. Six ruminally cannulated Holstein heifers (520 kg) were used in a split-plot design with treatments arranged as a 2 × 5 factorial. The main plot treatments were 0 or 12 g/d of l-Met arranged in a completely randomized design; three heifers received each main plot treatment throughout the entire experiment. Subplot treatments were 0, 10, 20, 30, and 40 g/d of GAA, with GAA treatments provided in sequence from lowest to highest over five 6-d periods. Treatments were infused continuously to the abomasum. Heifers were limit-fed twice daily a diet consisting of (dry matter basis) 5.3 kg/d rolled corn, 3.6 kg/d alfalfa hay, and 50 g/d trace-mineralized salt. Plasma Met increased (P < 0.01) when Met was supplemented, but it was not affected by supplemental GAA. Supplementing GAA linearly increased plasma arginine (% of total amino acids) and plasma concentrations of GAA and creatinine (P < 0.001). Plasma creatine was increased at all levels of GAA except when 40 g/d of GAA was supplemented with no Met (GAA-quadratic × Met, P = 0.07). Plasma homocysteine was not affected by GAA supplementation when heifers received 12 g/d Met, but it was increased when 30 or 40 g/d of GAA was supplemented without Met (GAA-linear × Met, P = 0.003); increases were modest and did not suggest a dangerous hyperhomocysteinemia. Urinary concentrations of GAA and creatine were increased by all levels of GAA when 12 g/d Met was supplemented; increasing GAA supplementation up to 30 g/d without Met increased urinary GAA and creatine concentrations, but 40 g/d GAA did not affect urine concentrations of GAA and creatine when no Met was supplemented. Overall, post-ruminal GAA supplementation increased creatine supply to cattle. A methyl group deficiency, demonstrated by modest increases in plasma homocysteine, became apparent when 30 or 40 g/d of GAA was supplemented, but it was ameliorated by 12 g/d Met.