Effect of dietary protein level on nitrogen metabolism in the growing bovine: II. Diffusion into and utilization of endogenous urea nitrogen in the rumen

Six Angus heifer calves (234 kg) were assigned to either a high (HP; 126.1 g N/d) or low (LP; 66.5 g N/d) protein intake to evaluate ruminal criteria associated with movement of blood urea-N (BUN)-derived NH3-N from the rumen wall into interior ruminal digesta. Calves received 4.8 kg DM/d of diets containing 30% cottonseed hulls and 70% cornsoybean meal in equal portions at 4-h intervals. Following single i.v. injections of 15N-urea, ruminal fluid was collected serially for 4 h postinjection from digesta located adjacent to the rumen wall (wall-proximate digesta; WPD) and from the center of the rumen digesta mass after manual agitation (center mixed digesta; CMD). Mean ruminal NH3-N (RAN) concentrations were higher (P less than .05) for HP than for LP, but were not affected (P greater than .05) by digesta sampling site. Ruminal urease activity was higher (P less than .05) for LP than for HP and tended (P = .14) to be higher for WPD than for CMD. Area under the 15N enrichment curve (AUC) ratios between sampling sites (WPD/CMD x 100) for RAN were greater (P less than .05) for LP than for HP. However, AUC ratios for bacterial N were not affected (P greater than .05) by protein level. Whereas BUN-derived 15NH3 appeared to thoroughly equilibrate with RAN in interior ruminal digesta with HP, there appeared to be a declining enrichment gradient for RAN from the rumen wall to the interior ruminal digesta with LP. Data are interpreted to suggest that bacteria at or near the rumen wall may preferentially utilize some BUN-derived NH3-N entering through the rumen wall in calves fed LP diets.     

Effects of barley grain processing and dietary ruminally degradable protein on urea nitrogen recycling and nitrogen metabolism in growing lambs

The objective of this study was to determine how interactions between dietary ruminally degradable protein (RDP) level and ruminally fermentable carbohydrate (RFC) alter urea N transfer to the gastrointestinal tract (GIT) and the utilization of this recycled urea N in rapidly growing lambs fed high-N diets. Four Suffolk ram lambs (34.8 +/- 0.5 kg of BW) were used in a 4 x 4 Latin square design with 21-d periods and a 2 x 2 factorial arrangement of dietary treatments. The dietary factors studied were 1) dry-rolled vs. pelleted barley as the principal source of RFC and 2) dietary levels of RDP of 60 vs. 70% (% of CP). All diets contained 28.8 g of N/kg of DM. Experimental diets were composed of 80% concentrate mixture and 20% barley silage (DM basis) and were fed twice daily at 0900 and 1700 as total mixed rations. Nitrogen balance was measured from d 15 to 20, and urea N kinetics were measured from d 15 to 19 using intrajugular infusions of [(15)N(15)N]-urea. Nitrogen intake (P = 0.001) and fecal (P = 0.002) and urinary (P = 0.03) N excretion increased as dietary RDP level increased, but the method of barley processing had no effect. Feeding dry-rolled compared with pelleted barley (P = 0.04) as well as feeding 60% RDP compared with 70% RDP (P = 0.04) resulted in a greater N digestibility. Whole-body N retention was unaffected (P >/= 0.74) by dietary treatment. Dietary treatment had no effect on endogenous production of urea N and its recycling to the GIT; however, across dietary treatments, endogenous production of urea N (45.8 to 50.9 g/d) exceeded N intake (42.3 to 47.9 g/d). Across dietary treatments, 30.6 to 38.5 g/d of urea N were recycled to the GIT, representing 0.67 to 0.74 of endogenous urea N production; however, 0.64 to 0.76 of urea N recycled to the GIT was returned to the ornithine cycle. In summary, although dietary treatment did not alter urea N kinetics, substantial amounts of hepatic urea N output were recycled to the GIT under the dietary conditions used in this study, and additional research is required to determine how this recycled urea N can be efficiently captured by bacteria within the GIT.     

Effect of dietary protein level on nitrogen metabolism in lambs: studies using 15N-nitrogen

Six wether lambs (31 kg) were randomly assigned to two treatments (three lambs/treatment): a high protein intake (HP; 21 g N/d) or a low protein intake (LP; 12 g N/d). Each lamb received 860 g/d dry matter (DM) of a pelleted diet (75% corn-soybean meal, 25% cottonseed hulls) offered hourly in 24 equal portions. Single injections of 15N-labelled compounds were made into the ruminal NH3-N and blood urea-N pools to measure the rate of flux through, and transfer of N between, these and the bacterial N pool. Total tract digestibilities of DM and N were lower (P less than .05) for the LP than the HP treatment. Abomasal flows of total, feed or bacterial N tended to be greater (P greater than .05) in lambs fed HP than LP. Lambs fed HP excreted more (P less than .01) urinary N, yet retained a greater (P less than .01) amount of N than lambs fed LP (6.2 vs 1.8 and 9.7 vs 4.1 g N/d, respectively). Pool size and production rate for both ruminal NH3-N and blood urea-N were greater (P less than .05) for the HP than LP treatment. Lambs consuming HP degraded more (P less than .05) blood urea-N in the gastro-intestinal tract (13.4 vs 6.9 g N/d); however, lambs fed LP degraded a greater (P less than .05) percentage of synthesized body urea-N (88.7 vs 71.8%). Ruminal NH3-N absorption was greater (P less than .01) for the HP than LP treatment (3.1 vs .5 g N/d). Although the percentage of bacterial N derived from ruminal NH3-N was similar (P greater than .05) between diets (51.1 vs 63.9), a greater (P less than .05) percentage of bacterial N was derived from blood urea-N in lambs fed LP than HP (77.1 vs 30.2%). Lambs fed LP incorporated a greater (P less than .10) amount of blood urea-N into bacterial N than lambs fed HP (5.5 vs 2.6 g N/d). These data are interpreted to suggest that blood urea-N may provide a substantial quantity of N for bacterial protein synthesis and, thus, may be an important source of protein in the deficient animal. In addition, urea recycling may play an important role in the recovery of ruminal NH3-N lost through absorption in animals fed a high level of protein.     

Effects of dietary energy level and protein source on nutrient digestion and ruminal nitrogen metabolism in steers.

Four Simmental steers with ruminal, duodenal, and ileal cannulas were used to examine effects of dietary forage: concentrate ratio and supply of ruminally degradable true protein on site of nutrient digestion and net ruminal microbial protein synthesis. Steers (345 kg) were fed ammoniated corn cob (high forage; HF)- or corn cob/ground corn/cornstarch (low forage; LF)-based diets supplemented with soybean meal (SBM) or a combination of corn gluten meal and blood meal (CB). Diets were fed at 2-h intervals with average DM intake equal to 2.2% of BW. Feeding LF vs HF increased (P less than .05) OM digestion (percentage of intake) in the stomach, small intestine, and total tract. Efficiency of microbial CP synthesis (EMCP; g of N/kg of OM truly fermented) decreased (P less than .05) for LF vs HF (24.1 vs 26.8), but microbial N and total N flows to the small intestine were similar (P greater than .05) between energy levels (average 112 and 209 g/d, respectively). Total N flows to the small intestine were 13.1% greater (P less than .05) for CB than for SBM because of increased (P less than .05) passage of nonmicrobial N. Feeding SBM vs CB increased (P less than .05) EMCP (27.3 vs 23.3) and microbial N flow to the small intestine (127.5 vs 112.5 g/d), but these increases were not likely due to increased ruminal concentrations of ammonia N (NH3 N). Decreased (P less than .05) incorporation of NH3 N into bacterial N and slower turnover rates of ruminal NH3 N for SBM vs CB suggest that direct incorporation of preformed diet components into cell mass increased when SBM was fed. Results of this study suggest that the inclusion of ruminally degradable protein in the diet may increase the supply of products from proteolysis and that this can increase EMCP and microbial protein flow to the small intestine.     

Influence of dietary protein on blood cholesterol and related metabolites of growing calves

Forty male, Holstein calves were utilized to evaluate effects of either low (12%) or high (25%) dietary protein on growth and serum metabolites, with a particular concern for serum cholesterol. All diets contained 19% acid detergent fiber and consisted of corn silage, alfalfa hay and grain mixtures. Average concentration of total serum cholesterol was depressed (P = .001) for calves on the high-protein diets (58.8 mg/dl) as compared with their low-protein counterparts (67.9 mg/dl). Calves fed the high-protein diet had lower (P = .002) free serum cholesterol but elevated high-density lipoprotein cholesterol as a percentage of total cholesterol. Calves receiving the high-protein diets showed increases (P = .206) in lecithin cholesterol acyltransferase activity compared with low-protein counterparts. Serum urea nitrogen was increased (P = .001) for calves fed high-protein diets, while serum protein and glucose remained unchanged. Calves fed the high-protein diets consumed less dry matter with improved (P = .09) growth efficiency (gain/feed).     

Influence of dietary protein concentrations on performance and nitrogen repletion in stressed calves

Three trials were conducted to determine the influence of dietary CP concentration on health and performance of market-transport-stressed feeder calves (Exp. 1 and 2) and on repletion of nutrients lost during a 3-d feed and water deprivation period in steers fed at maintenance energy intake (Exp. 3). In Exp. 1 (84 calves) and 2 (256 calves), feeder calves averaging 184 kg were transported from Tennessee to Texas. In Exp. 1, calves were fed receiving diets containing either 12 or 16% CP. In Exp. 2, calves were fed diets containing 12 or 16% CP and .8 or 1.3% potassium in a 2 x 2 factorial arrangement of treatments. In Exp. 3, four Hereford steers averaging 253 kg were used in an N balance trial. Steers were deprived of feed and water for 3 d and then were limit-fed (1 x maintenance energy requirements) diets calculated to meet 100, 120, 140 or 160% of CP maintenance requirements for 14 d in a 4 x 4 Latin square design. In Exp. 1, calves fed the 16% CP diet had faster (P less than .05) daily gains and higher (P less than .10) feed consumption than calves fed the 12% CP diet during the first 14 d. In Exp. 2, calf performance was not affected by diet CP or K content. Calves fed the 16CP-1.3K diet had lower (P less than .10) mortality than calves on the remaining treatments. In Exp. 3, N balance and serum urea N increased linearly (P less than .05) with increasing dietary CP. Results of these studies are interpreted to indicate that the CP requirement (g/d) of market-transport-stressed feeder calves is similar to requirements of nonstressed calves; however, the CP concentration of the diet of stressed calves may need to be increased when feed intakes are low.     

Energy metabolism and protein balance in growing rats fed different levels of dietary fibre and protein.

A study was performed to investigate the effect of different levels of dietary fibre (DF) and dietary protein on visceral organ size, digestibility, nitrogen balance and energy metabolism in rats. Thirty-six male Wistar rats, initial body weight about 76 g were used in a factorial design consisting of three levels of DF (low, 100 g/kg DM: medium, 250 g/kg DM and high, 290 g/kg DM) and two levels of dietary protein (low, 120 g/kg DM and high, 223 g/kg DM). The added fibre source was soybean hulls and Danish fish meal was used as sole source of dietary protein. Measurements of gas-exchange were done on six rats (one group) while urine and faeces were collected individually. The ratio of food/empty body gain increased (P<0.05) with increasing DF and decreasing levels of dietary protein. The weight of the digestive tract was larger (P<0.05) in rats fed the high fibre diet than in those fed the low fibre diet. The digestibility of nutrients and energy decreased linearly with increasing level of soybean fibre (P<0.05). An increased intake of DF was associated with a concomitant loss of protein and energy to faeces. The microbial degradation of NSP and other unabsorbed carbohydrates caused considerably changes in N metabolism of the colon. In rats fed the low protein diets increased levels of DF decreased N excretion in urine and increased N excretion in faeces, while the ratio of retained/digested protein remained constant. When rats were fed the high protein diet protein retention dropped in response to DF both absolute and relative to digested amount, indicating that energy intake could be a limiting factor. Heat production as a percentage of metabolizable energy (HP/ME) was higher (P<0.05) in rats fed the low protein diet than in rats fed the high protein diet, but no significant difference was found among DF levels.     

Protein and amino acid metabolism in the intestinal tract of growing young bulls. 7. Protein estimation and evaluation of protein supply in growing ruminants by the apparently digestible total protein in the duodenum based on rations

The following values were derived from experimental studies of the crude protein (CP) and amino acid (AA) metabolism of growing fattening bulls, the detailed results of which were described in installments 1-6, for the ascertainment of the content of apparently digestible total protein at the duodenum (ADTPD): a) partial method: Degradation quotaCP = NPN + 0.69 X g pure protein/CP; g AD feed PD = g CP (1 -decomposition quotaCP) X 0.72 or g AD feed PD = 0.31 X g pure protein X 0.72; g ADBPDE = 0.149 X g AD org. matter X 0.80 X 0.72 (ADBPDE = apparently digestible pure bacteria protein at the duodenum, dependent on energy release in the reticulo-rumen; g ADBPDN = g CP X (degradation quotaCP) X 0.80 X 0.72 (ADBPDN = apparently digestible pure bacteria protein at the duodenum, dependent on CP supply of rumen microbes); g AD total PD = g AD feed PD + ADBPDE g AD total PD = g AD feed PD + ADBPDN; b) summarizing method: AD total PD = 0.429 + 10.9 X 0.80/CP in % AD org. matter X (CP intake) X 0.72; AD total PD = 0.196 + 16.6 X 0.80/pure protein % AD org. m. X pure pr. intake X 0.72. The requirement of AD total PD was calculated under consideration of the endogenous urine N, skin N and hair N losses as well as protein retention per kg live weight assuming a utilization of 0.70 for this partial performance. The practical application of this balance method is demonstrated in comparison to the digestible CP-system.     

Effect of imbalance between energy and nitrogen supplies on microbial protein synthesis and nitrogen metabolism in growing double-muscled Belgian Blue bulls

Six double-muscled Belgian Blue bulls (initial weight: 345 +/- 16 kg) with cannulas in the rumen and proximal duodenum were used in two juxtaposed 3 x 3 Latin squares to study the effect of a lack of synchronization between energy and N in the rumen on microbial protein synthesis and N metabolism by giving the same diet according to three different feeding patterns. The feed ingredients of the diet were separated into two groups supplying the same amount of fermentable OM (FOM), but characterized by different levels of ruminally degradable N (RDN). The first group primarily provided energy for the ruminal microbes (14.6 g of RDN/kg of FOM), and the second provided N (33.3 g of RDN/kg of FOM). These two groups were fed to the bulls simultaneously or alternately with the aim of creating three different time periods of imbalance (0, 12, or 24 h) between energy and N supplies in the rumen. The introduction of imbalance affected neither microbial-N flow at the duodenum (P = 0.65) nor efficiency of growth (P = 0.69), but decreased (P = 0.016) the NDF degradation in the rumen 12.2% for a 12-h period of imbalance. N retention was not affected by imbalance (P = 0.53) and reached 57.8, 58.5, and 54.7 g/d, respectively, for 0-, 12- and 24-h imbalance. It seems that the introduction of an imbalance of 12 or 24 h between energy and N supplies for the ruminal microbes by altering the feeding pattern of the same diet does not negatively influence microbial protein synthesis or N retention by the animal. Nitrogen recycling in the rumen plays a major role in regulating the amount ofruminally available N and allows for continuous synchronization of N- and energy-yielding substrates for the microorganisms in the rumen. Therefore, a lack of synchronization in the diet between the energy and N supplies for the ruminal microbes is not detrimental to their growth or for the animal as long as the nutrient supply is balanced on a 48-h basis. Thus, these dietary feeding patterns may be used under practical feeding conditions with minimal effect on the performance of ruminant animals.     

Effects of dietary energy level and supplemental protein source on performance of growing steers and nutrient digestibility and nitrogen balance in lambs

Four experiments were conducted to evaluate three crude protein (CP) sources (urea, U; soybean meal, SBM; corn gluten meal, CGM) in diets based on corn silage (high energy) or grass hay (low energy). In Exp. 1 and 2, growing steers were fed all combinations of energy and protein source at 10.5 or 12% CP. Steers fed high energy diets or 12% CP had improved (P less than .05) daily gains and feed:gain over 84 d. Protein source had no effect (P greater than .05) on performance except that steers fed U consumed more (P less than .05) feed than those fed CGM. Steers were fed experimental diets to a common weight and switched to an 85% concentrate diet for finishing. During finishing, steers fed low energy diets in the growing period consumed more (P less than .05) feed and had increased (P less than .05) feed:gain compared with those fed high energy diets. Growing lambs were fed the same diets as steers. At 10.5% CP, lambs fed high energy diets had higher (P less than .05) digestibilities of dry matter (DM), organic matter (OM), nitrogen (N) and fiber components, and retained more (P less than .05) N. For lambs on 12% CP, high energy diets had higher (P less than .05) DM and OM digestibilities and lower (P less than .05) N digestibilities. At 12% CP, energy level had no effect (P greater than .05) on N retained. Protein source had no effect (P greater than .05) on N retention. There appeared to be no advantage in supplementing with ruminally undegradable proteins, i.e. CGM, in these experiments.     

Effect of dietary medium chain triglyceride on protein and energy utilisation in growing chicks.

1. Protein, fat and energy retentions of chicks fed on diets containing medium chain triglyceride (MCT) and long chain triglyceride (LCT), at 100 and 200 g fat/kg diet, were investigated. Maize oil was used as the LCT source. The MCT used was glyceryl tricaprylate. 2. Body weight gain and food intake were decreased with the diets containing MCT and these effects were greater at the higher fat concentration. Protein retention was also reduced by dietary MCT, although the efficiency of protein utilisation (protein retained/protein intake) was not altered. 3. The values for fat and energy retentions were significantly lower in the chicks fed on the MCT-supplemented diets than in those receiving the LCT-containing diets. Dietary ME values and efficiencies for energy utilisation (energy retained/ME intake) were also reduced by dietary MCT.     

Effect of dietary crude protein level and degradability on ruminal fermentation and nitrogen utilization in lactating dairy cows

The objectives of this experiment were to investigate the effects of two ruminally degradable protein (RDP) levels in diets containing similar ruminally undegradable protein (RUP) and metabolizable protein (MP) concentrations on ruminal fermentation, digestibility, and transfer of ruminal ammonia N into milk protein in dairy cows. Four ruminally and duodenally cannulated Holstein cows were allocated to two dietary treatments in a crossover design. The diets (adequate RDP [ARDP] and high RDP [HRDP]), had similar concentrations of RUP and MP, but differed in CP/RDP content. Ruminal ammonia was labeled with 15N and secretion of tracer in milk protein was determined for a period of 120 h. Ammonia concentration in the rumen tended to be greater (P = 0.06) with HRDP than with ARDP. Microbial N flow to the duodenum, ruminal digestibility of dietary nutrients, DMI, milk yield, fat content, and protein content and yield were not statistically different between diets. There was a tendency (P = 0.07) for increased urinary N excretion, and blood plasma and milk urea N concentrations were greater (P = 0.002 and P = 0.01, respectively) with HRDP compared with ARDP. Milk N efficiency was decreased (P = 0.01) by the HRDP diet. The cumulative secretion of ammonia 15N into milk protein, as a proportion of 15N dosed intraruminally, was greater (P = 0.003) with ARDP than with HRDP. The proportions of bacterial protein originating from ammonia N and milk protein originating from bacterial or ammonia N averaged 43, 61, and 26% and were not affected by diet. This experiment indicated that excess RDP in the diet of lactating dairy cows could not be efficiently utilized for microbial protein synthesis and was largely lost through urinary N excretion. At a similar MP supply, increased CP or RDP concentration of the diet would result in decreased efficiency of conversion of dietary N into milk protein and less efficient use of ruminal ammonia N for milk protein syntheses.     

Effect of dietary dry matter intake on endogenous nitrogen flows in growing lambs

The effect of dietary dry matter intake (DMI) on endogenous nitrogen (N) flows at different part of the digestive tract of growing lambs was determined using a ¹⁵N isotope dilution technique. Three Kazakh male lambs (30 ± 2.75 kg of body weights and 4 months old, average daily gain 200 g/day) were fitted with ruminal, duodenal and ileal cannulae and raised in metabolic cage individually. The experiment was conducted in a 3 × 3 Latin square design with three lambs, three DMI levels (1100, 920 and 736 g/day respectively) and three periods. Each period lasted 18 days, consisting of 10 days for adaptation, 8 days for the continuous infusion of l ‐[¹⁵N]leucine, during which the intestinal flow of N and ¹⁵N enrichment were determined. The total endogenous secretions in the forestomach (S_fs) were decreased (p = 0.0512) with increased level of DMI. On the contrary, endogenous nitrogen (EN) secretions into the small intestine (S_i) increased (p = 0.0249) significantly with the high level of DMI (HI) group compared with that of low level of DMI (LI). Total absorption from forestomach was reduced (p = 0.0121) with increased level of DMI, whereas total absorption from small intestine for HI group increased (p = 0.0116) significantly compared with that of LI treatment. The real digestibility of N in the rumen accompanied with the increase in feed intake is decreased (p = 0.081). In contrast, there were no effects of DMI level on the computed real digestibility of N across both small intestine and whole tract. The results of this study indicate that the total flows of EN at duodenum may be unaffected by the level of DMI; however, the EN flow at ileal level increased from 12% to 37% with the increase in DMI level, corresponding to 33% of total N flow at ileum.