The effect of CP concentration in the diet on urea kinetics and microbial usage of recycled urea in cattle: a meta-analysis.

In ruminants, urea recycling is considered an evolutionary advantage. The amount of urea recycled mainly depends of the nitrogen (N) intake and the amount of organic matter (OM) digested in the rumen. Because recycled N contributes to meeting microbial N requirements, accurate estimates of urea recycling can improve the understanding of efficiency of N utilization and N losses to the environment. The objective of this study was to evaluate urea kinetics and microbial usage of recycled urea N in ruminants using a meta-analytical approach. Treatment mean values were compiled from 25 studies with ruminants (beef cattle, dairy cows and sheep) which were published from 2001 to 2016, totalling 107 treatment means. The data set was analyzed according to meta-analysis techniques using linear or non-linear mixed models, taking into account the random variations among experiments. Urea N synthesized in the liver (UER) and urea N recycled to the gut (GER) linearly increased (P<0.001) as N intake (g/BW0.75) increased, with increases corresponding to 71.5% and 35.2% of N intake, respectively. The UER was positively associated (P<0.05) with dietary CP concentration and the ratio of CP to digestible OM (CP:DOM). Maximum curvature analyses identified 17% dietary CP as the point where there was a prominent increase in hepatic synthesis of urea N, likely due to an excess of dietary N leading to greater ammonia absorption. The GER:UER decreased with increasing dietary CP concentration (P<0.05). At dietary CP⩾19%, GER:UER reached near minimal values. The fraction of UER eliminated as urinary urea N and the contribution of urea N to total urinary N were positively associated with dietary CP (P<0.05), both reaching values near the plateau when dietary CP was 17%. The fractions of GER excreted in the feces and utilized for anabolism decreased, whereas the fraction of GER returned to the ornithine cycle increased with dietary CP concentration (P<0.05). Recycled urea N assimilated by ruminal microbes (as a fraction of GER) decreased as dietary CP and CP:DOM increased (P<0.05). The efficiency of microbial assimilation of recycled urea N was near plateau values at 194 g CP/kg DOM. The models obtained in this study contribute to the knowledge on N utilization, and they could be used in feeding models to predict urea recycling and thus to improve formulation of diets to reduce N losses that contribute to air and water pollution.

Efficiency of lysine utilization by growing steers.

This study evaluated the efficiency of Lys utilization by growing steers. Five ruminally cannulated Holstein steers (165 ± 8 kg) housed in metabolism crates were used in a 6 × 6 Latin square design; data from a sixth steer was excluded due to erratic feed intake. All steers were limit fed (2.46 kg DM/d), twice daily, diets low in RUP (81% soybean hulls, 8% wheat straw, 6% cane molasses, and 5% vitamins and minerals). Treatments were 0, 3, 6, 9, 12, and 15 g/d of Lys continuously abomasally infused. To prevent AA other than Lys from limiting performance, a mixture providing all essential AA to excess was continuously abomasally infused. Additional continuous infusions included 10 g urea/d, 200 g acetic acid/d, 200 g propionic acid/d, and 50 g butyric acid/d to the rumen and 300 g glucose/d to the abomasum. These infusions provided adequate ruminal ammonia and increased energy supply without increasing microbial protein supply. Each 6-d period included 2 d for adaptation and 4 d for total fecal and urinary collections for measuring N balance. Blood was collected on d 6 (10 h after feeding). Diet OM digestibility was not altered ( ≥ 0.66) by treatment and averaged 73.7%. Urinary N excretion was decreased from 32.3 to 24.3 g/d by increasing Lys supplementation to 9 g/d, with no further reduction when more than 9 g/d of Lys was supplied (linear and quadratic, < 0.01). Changes in total urinary N excretion predominantly were due to changes in urinary urea N. Increasing Lys supply from 0 to 9 g/d increased N retention from 21.4 to 30.7 g/d, with no further increase beyond 9 g/d of Lys (linear and quadratic, < 0.01). Break-point analysis estimated maximal N retention at 9 g/d supplemental Lys. Over the linear response surface of 0 to 9 g/d Lys, the efficiency of Lys utilization for protein deposition was 40%. Plasma urea N tended to be linearly decreased ( = 0.06) by Lys supplementation in agreement with the reduction in urinary urea N excretion. Plasma concentrations of Lys linearly increased ( < 0.001), but Leu, Ser, Val, and Tyr ( ≤ 0.02) were linearly reduced by Lys supplementation, likely reflecting increased uptake for protein deposition. In our model, Lys supplementation promoted significant increases in N retention and was maximized at 9 g/d supplemental Lys with an efficiency of utilization of 40%.

Effect of ruminal ammonia supply on lysine utilization by growing steers.

Six ruminally cannulated Holstein steers (202 ± 15 kg) were used to study the effects of ruminal ammonia loading on whole-body lysine (Lys) utilization. Steers were housed in metabolism crates and used in a 6 × 6 Latin square design. All steers received 2.52 kg DM/d of a diet (10.1% CP) containing 82% soybean hulls, 8% wheat straw, 5% cane molasses, and 5% vitamins and minerals, and 10 g/d of urea (considered to be part of the basal diet) was ruminally infused continuously to ensure adequate ruminal ammonia concentrations. All steers were ruminally infused continuously with 200 g/d of acetic acid, 200 g/d of propionic acid, and 50 g/d of butyric acid and abomasally infused with 300 g/d of glucose continuously to increase energy supply without increasing microbial protein supply. Steers were also abomasally infused continuously with an excess of all essential AA except Lys to ensure that Lys was the only limiting AA. Treatments were arranged as a 3 × 2 factorial with 3 levels of urea (0, 40, or 80 g/d) continuously infused ruminally to induce ammonia loading and 2 levels of Lys (0 or 6 g/d) continuously infused abomasally. Treatments did not affect fecal N output ( = 0.37). Lysine supplementation decreased ( < 0.01) urinary N excretion from 51.9 g/d to 44.3 g/d, increased ( < 0.01) retained N from 24.8 to 33.8 g/d, increased ( < 0.01) plasma Lys, and decreased ( ≤ 0.05) plasma serine, tyrosine, valine, leucine, and phenylalanine. Lysine supplementation also tended ( = 0.09) to reduce plasma urea-N. Urea infusions linearly increased ( = 0.05) retained N (27.1, 29.3, and 31.5 g/d) and also linearly increased ( < 0.01) urinary N excretion (31.8, 48.1, and 64.4 g/d), urinary urea (21.9, 37.7, and 54.3 g/d), urinary ammonia (1.1, 1.4, and 1.9 g/d), and plasma urea (2.7, 4.0, and 5.1 mM), and linearly decreased plasma alanine ( = 0.04) and plasma glycine ( < 0.01). Assuming that retained protein is 6.25 × retained N and contains 6.4% Lys, the incremental efficiencies of infused Lys utilization were 51%, 59%, and 69% for steers receiving 0, 40, and 80 g/d of urea, respectively, indicating that ruminal ammonia loads may improve the efficiency of Lys utilization. This is supported by observed increases in whole body-protein deposition in response to ammonia loading of our steers that were, by design, Lys deficient.

Effects of Supplements with Different Protein Contents on Nutritional Performance of Grazing Cattle During the Rainy Season.

The objective of this study was to evaluate the effects of supplements with different crude protein (CP) contents on grazing cattle intake, digestibility, ruminal fermentation pattern, and nitrogen (N) metabolism characteristics during the rainy season. Five ruminal and abomasal cannulated Holstein×Zebu steers (296 kg body weight, BW) were used in a 5×5 Latin square design. The animals grazed five signal grass paddocks (0.34 ha). The five treatments evaluated were: Control (no supplement) and 1.0 g of supplement/kg BW with 0, 330, 660, and 1,000 g of CP/kg as-fed. The supplement was composed of starch, soybean meal, urea, and ammonium sulphate. There was a positive linear effect (p≤0.033) of the CP content in the supplements on the organic matter (OM), CP, and digested OM intakes. The provision of supplements did not increase (p≥0.158), on average, total and ruminal digestibilities of OM and CP. However, the increase in CP content in the supplements caused a positive linear effect (p≤0.018) on ruminal digestibilities of OM and CP. Additionally, a quadratic effect of the CP contents of the supplements were observed (p = 0.041) for the ruminal digestibility of neutral detergent fiber corrected for ash and protein, with the highest estimate obtained with the CP content of 670 g/kg. The supply of supplements increased (p<0.001) the ruminal ammonia N concentration, which also changed linearly and positively (p<0.001) according to increase in CP content in the supplements. The apparent N balance and relative N balance (g/g N intake) were not, on average, changed (p≥0.164) by the supplements supply. However, both showed a tendency of a linear increase (p≤0.099) with increasing supplement CP content. The supplements increased (p = 0.007) microbial N production in the rumen, which also changed linearly and positively (p = 0.016) with increasing supplement CP content. In conclusion, protein supplementation in grazing cattle during the rainy season, while stimulating voluntary forage intake, results in higher efficiency of N utilization when compared to energy supplementation. This is a possible response to increased microbial protein synthesis in the rumen and improved N status in the animal body.

Effects of varying ruminally undegradable protein supplementation on forage digestion, nitrogen metabolism, and urea kinetics in Nellore cattle fed low-quality tropical forage.

Effects of supplemental RDP and RUP on nutrient digestion, N metabolism, urea kinetics, and muscle protein degradation were evaluated in Nellore heifers () consuming low-quality signal grass hay (5% CP and 80% NDF, DM basis). Five ruminally and abomasally cannulated Nellore heifers (248 ± 9 kg) were used in a 5 × 5 Latin square. Treatments were the control (no supplement) and RDP supplementation to meet 100% of the RDP requirement plus RUP provision to supply 0, 50, 100, or 150% of the RUP requirement. Supplemental RDP (casein plus NPN) was ruminally dosed twice daily, and RUP supply (casein) was continuously infused abomasally. Jugular infusion of [NN]-urea with measurement of enrichment in urine was used to evaluate urea kinetics. The ratio of urinary 3-methylhistidine to creatinine was used to estimate skeletal muscle protein degradation. Forage NDF intake (2.48 kg/d) was not affected ( ≥ 0.37) by supplementation, but supplementation did increase ruminal NDF digestion ( < 0.01). Total N intake (by design) and N retention increased ( < 0.001) with supplementation and also linearly increased with RUP provision. Urea entry rate and gastrointestinal entry rate of urea were increased by supplementation ( < 0.001). Supplementation with RUP linearly increased ( = 0.02) urea entry rate and tended ( = 0.07) to linearly increase gastrointestinal entry rate of urea. Urea use for anabolic purposes tended ( = 0.07) to be increased by supplementation, and RUP provision also tended ( = 0.08) to linearly increase the amount of urea used for anabolism. The fraction of recycled urea N incorporated into microbial N was greater ( < 0.001) for control (22%) than for supplemented (9%) heifers. Urinary 3-methylhistidine:creatinine of control heifers was more than double that of supplemented heifers ( < 0.001). Control heifers reabsorbed a greater ( < 0.001) fraction of urea from the renal tubule than did supplemented heifers. Overall, unsupplemented heifers had greater mobilization of AA from myofibrillar protein, which provided N for urea synthesis and subsequent recycling. Supplemental RUP, when RDP was supplied, not only increased N retention but also supported increased urea N recycling and increased ruminal microbial protein synthesis.

Effect of protein supplementation on ruminal parameters and microbial community fingerprint of Nellore steers fed tropical forages.

In tropical regions, protein supplementation is a common practice in dairy and beef farming. However, the effect of highly degradable protein in ruminal fermentation and microbial community composition has not yet been investigated in a systematic manner. In this work, we aimed to investigate the impact of casein supplementation on volatile fatty acids (VFA) production, specific activity of deamination (SAD), ammonia concentration and bacterial and archaeal community composition. The experimental design was a 4×4 Latin square balanced for residual effects, with four animals (average initial weight of 280±10 kg) and four experimental periods, each with duration of 29 days. The diet comprised Tifton 85 (Cynodon sp.) hay with an average CP content of 9.8%, on a dry matter basis. Animals received basal forage (control) or infusions of pure casein (230 g) administered direct into the rumen, abomasum or divided (50 : 50 ratio) in the rumen/abomasum. There was no differences (P>0.05) in ruminal pH and microbial protein concentration between supplemented v. non-supplemented animals. However, in steers receiving ruminal infusion of casein the SAD and ruminal ammonia concentration increased 33% and 76%, respectively, compared with the control. The total concentration of VFA increased (P0.05) in species richness and diversity of γ-proteobacteria, firmicutes and archaea between non-supplemented Nellore steers and steers receiving casein supplementation in the rumen. However, species richness and the Shannon-Wiener index were lower (P<0.05) for the phylum bacteroidetes in steers supplemented with casein in the rumen compared with non-supplemented animals. Venn diagrams indicated that the number of unique bands varied considerably among individual animals and was usually higher in number for non-supplemented steers compared with supplemented animals. These results add new knowledge about the effects of ruminal and postruminal protein supplementation on metabolic activities of rumen microbes and the composition of bacterial and archaeal communities in the rumen of steers.