Nucleic acid synthesis from blood urea 15N in the sheep

In experiments on 4 sheep fed on a low protein diet [6.2 g N/day] and given a single i.v. dose of 15N-labelled urea [15 mg 15N/kg body mass], the authors found that, from 0.5 to 6 h, mean 15N incorporation rose progressively in the total rumen fluid nitrogen from 0.23 to 0.44 at. % 15N and in the rumen bacterial nitrogen from 0.11 to 0.51 at. % 15N. Up to 3 h, total nitrogen enrichment was greater (0.5 at. % 15N) than enrichment of bacterial nitrogen (0.28 at. % 15N), but from 3 to 6 h there was little difference between them. The mean 15N values in the nucleic acids isolated from rumen fluid bacteria in samples collected 3 and 6 hours after injecting labelled urea into the blood were 0.15 and 0.19 at. % 15N respectively, in nucleic acids isolated from the liver 0.042 and 0.04 at. % 15N, in the total rumen bacterial nitrogen 0.28 and 0.51 at. % 15N and in the total liver nitrogen 0.11 and 0.11 at. % 15N. It is concluded from the results that blood urea nitrogen is utilized for synthesis of the total nitrogenous substances of the sheep's rumen bacteria and liver far more intensively than for synthesis of the nucleic acids isolated from them. At the same time, it is utilized more intensively for nucleic acid synthesis in the rumen bacteria than in the liver.     

Towards non-invasive methods to determine the effect of treatment of soya-bean meal on lysine availability in dairy cows

Lysine (Lys) availability in three different soya-bean meal (SBM) products was determined using the following techniques: whole body (WB) net flux of Lys, digestible Lys (duodenal flow × intestinal digestibility) and the plasma Lys response curve method of Rulquin and Kowalczyk (2003). Four multiparous Holstein cows (173 days in milk) were equipped with ruminal and duodenal cannulas and used in a 4 × 4 Latin square experiment with 14-day periods. The animals were fed either solvent-extracted SBM (SE), expeller-processed SBM (EP) or lignosulphonate-treated SBM (LS) at 23% of the diet dry matter (DM). The fourth treatment (SE70) consisted of a continuous infusion of Lys (70 g/day) into the omasum of cows fed the SE diet. Chromium(III) oxide was included as a digesta marker in order to determine the duodenal flow of amino acids (AA). On day 12 of each experimental period, six blood samples were collected to determine plasma Lys concentrations. Immediately after that, a pulse dose of L-[2-15N] Lys was administered in the jugular vein. Jugular blood samples were then collected at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 13, 16, 19, 25 and 31 min after the injection to determine 15N Lys enrichment. On each of days 13 and 14, eight digesta samples were collected and pooled by period. Amongst the diets of SBM (SE, EP, LS), no differences were observed for duodenal Lys flow or digestible Lys. Duodenal flow of microbial N with SE was numerically higher, compared with EP and LS, indicating enhanced duodenal supply of microbial Lys for this diet, and this may have compensated for the additional Lys derived from undegradable protein in rumen-protected SBM products (EP and LS). The use of the plasma response curve method as well as the measurement of WB Lys flux also revealed no differences in Lys availability among the SBM products. The WB flux method resulted in 100% post-ruminal recovery of the Lys infused with diet SE70 compared with the control diet SE, which indicates that the method is reliable for determining Lys availability. The Lys flux approach not only allows for the estimation of intestinally available essential AA but also it avoids the use of cannulated animals.     

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/BW^0.75) 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.     

Influence of urea calcium mixture supplementation on ruminal fermentation characteristics of beef cattle fed on concentrates containing high levels of cassava chips and rice straw

The purpose of this study was to evaluate the effects of various N sources in concentrates containing high levels of cassava chips, with rice straw as the basal forage, on rumen ecology, rumen microbial counts, microbial crude (CP) protein synthesis, and digestibility of nutrients. Four ruminally fistulated crossbred (Brahman × native) beef steers with initial body weight (BW) of 400 ± 40.2 kg were randomly assigned according to a 4 × 4 Latin square design. The dietary treatments were different sources of N in the concentrates and were: T1 = urea (control; urea); T2 = soybean meal (SBM); T3 = urea CaCl₂ mixture (U-Cal); T4 = urea CaSO₄ mixture (U-Cas). All steers were kept in individual pens and supplemented with concentrate at 5 g/kg of BW daily. The experiment was 4 periods, and each lasted 21 d. During the first 14 d, all steers were fed their respective diets ad libitum and for during the last 7 d, they were moved to metabolism crates for total urine and fecal collection. Dry matter intake ranged from 9.8 to 10.5 kg daily and was not altered by diet, while digestibility of NDF differed among treatments and was highest with U-Cas supplementation (P<0.05). Ruminal NH₃ N and plasma urea N with U-Cal, U-Cas, and SBM diets were lower compared with the urea supplemented group (P<0.05). Ruminal volatile fatty acid concentrations were not altered by treatments. Total viable, and cellulolytic bacteria, differed among treatments and were highest with U-Cas (9.1 × 10¹¹, and 4.0 × 10⁹ cfu/mL, respectively). In addition, efficiency of rumen microbial CP synthesis based on organic matter (OM) truly digested in the rumen was increased by SBM or U-Cal supplementation, and was highest with U-Cas supplementation (18.2 g of N/kg of OM truly digested in the rumen). Supplementation of U-Cas to a concentrate containing a high level of cassava chips improved rumen ecology and microbial CP synthesis in beef cattle, suggesting that urea calcium mixtures can replace soybean meal or urea in beef cattle diets without adverse affects on rumen fermentation and other rumen parameters.     

Evaluation of different markers for determination of the microbial nitrogen flow into the duodenum of dairy cows

2.6‐Diaminopimelic acid (DAPA), ribonucleic acid (RNA), ¹⁵N, D‐alanine (D‐ALA) and the amino acid profiles (AAP) were compared as microbial markers for determination of the microbial protein synthesis in the rumen. Three dairy cows (Schwarzbuntes Milchrind, LW 602 kg), each fitted with a rumen cannula and a re‐entrant cannula in the proximal duodenum, were offered four isoenergetic and isonitrogenous diets (mean daily intake 15.0 ± 0.45 kg DM; forage: concentrate = 50:50) in a periodic experiment. The diets contained soyabean extracted meal, meat and bone meal, pea meal and dried clover as major sources of protein. On the 4th day after administration of 9 g ¹⁵N‐labelled urea (95 atom‐% ¹⁵N‐excess) per day, samples of rumen fluid and duodenal digesta were obtained 3 h after feeding. The bacteria were isolated by differential centrifugation. Bacteria harvested from the rumen had significantly higher ¹⁵N enrichment and D‐ALA: N ratio than ‘duodenal’ bacteria. However, DAPA: N ratio was higher in ‘duodenal’ bacteria compared to rumen bacteria. There were no differences in RNA: N ratio between rumen and ‘duodenal’ bacteria. The source of the bacteria in the digestive tract has an influence on the ratio of microbial N: total N, especially when ¹⁵N, AAP, DAPA and D‐ALA but not RNA were used as markers. The most reproducible method was D‐ALA (C.V. 4.7 for rumen and 6.8 for ‘duodenal’ bacteria) followed by ¹⁵N (10.8 resp. 4.8) and RNA (9.7 resp. 8.2). The results obtained with ¹⁵N and D‐ALA agreed closely at the same source of bacteria. The RNA method reached the level of these markers (¹⁵N, D‐ALA) when the bacteria were isolated from the duodenum. It is concluded that D‐ALA (bacteria isolated from rumen and duodenum) and also ¹⁵N (bacteria isolated from duodenum) were the best markers for estimation of the microbial protein synthesis.     

Availability of nitrogen from 15N-labelled Azolla pinnata and urea to flooded rice

In view of the recently generated interest in Azolla and the high cost of N fertilizers, this field study was aimed at measuring the availability of Azolla-N applied in two split application in comparison to urea-N. Azolla was cultivated and labelled with ¹⁵N isotope in the field. A total of about 60 kg N ha^-1 was applied as Azolla, urea or Azolla and urea in combination, in two equal splits at transplanting and at maximum tillering, i.e. 30 days after transplanting (30 DAT).The recovery by the crop of Azolla-N applied at 30 DAT was significantly higher than that applied at transplanting, viz. 30.2% and 20.2%, respectively. The recoveries of urea-N applied at the same stages were similarly low, viz. 22.5% at transplanting and 38.6% at 30 DAT. Total recoveries of fertilizer N at the time of harvest were 26.8% from Azolla, 30.7% from urea applied in the same two splits and 49.1% from urea applied in locally recommended three splits. Recoveries of labelled Azolla-N in succeeding rice crop were twice higher than those of labelled urea-N. The recoveries ranged from 1.9 to 2.1% from urea-N and 4.0 to 4.9% from Azolla-N. There were no differences in residual ¹⁵N recovery in the succeeding crop between Azolla and urea either applied at transplanting or at 30 DAT.     

Investigations on the effect of the ruminal N-balance on rumen metabolism, urea content in blood serum and milk as well as some liver parameters of lactating cows

Seven dairy cows fitted with ruminal and duodenal cannulae were used to investigate the influence of the amount of ruminally available N (Ruminal N-Balance, RNB) on the rumen metabolism and to answer the question on the lowest N-amount in the rumen, without negative effects on the fermentation. Animals were fed a ration on the basis of 7.9 kg corn silage and 7.2 kg concentrates related to dry matter, intended to meet the animals mean NEL and protein requirements. RNB amounted to -0.6 g/MJ ME in the basis ration. The other 3 rations were adjusted to RNB-values of -0.3, 0 and + 0.3 g/MJ ME by urea supplements in the concentrates. The increase in RNB resulted in higher NH3-N concentrations in the rumen fluid and in the duodenal digesta and higher urea concentrations in the blood and milk. The significantly highest amount of protein at the duodenum was detected when RNB showed an equilibrium (RNB = 0). The efficiency of microbial protein (MP) synthesis (gMP/kg fermented organic matter) was the same, g MP/d and g MP/MJ ME were significantly lower with RNB = -0.6g/MJ ME as compared to RNB = 0. The group with thelowest RNB showed the highest level of feedprotein degradation as well as the lowest organic matter, NDF and ADF fermentation. An effect on cholesterol, total bilirubin and gammaGT due to different RNB was not detected. The activities of GLDH and AST were highest when the RNB was -0.6 g/MJ ME. From the results, it can be concluded that significantly negative effects on rumen fermentation occur when RNB-values are below -0.3 g/MJ ME. However, a positive RNB did not increase t he degradation and synthesis capacity of the rumen micro-organisms as compared to RNB = 0.     

抑制剂和猪粪对尿素氮在稻田土壤中转化的影响

为了阐明稻田土壤中尿素在配施抑制剂和猪粪的情况下不同形态氮的响应特征,探究不同管理措施下稻田土壤氮素保持和供给能力。本研究采用¹⁵N标记尿素进行盆栽试验,设置不施肥(CK)、猪粪(M)、尿素(N)、猪粪+尿素(NM)、尿素+抑制剂(NI)和尿素+抑制剂+猪粪(NIM)6个处理。抑制剂选用脲酶抑制剂(PPD+NBPT)和硝化抑制剂(DMPP)组合,测定返青期、分蘖期和成熟期土壤氮库的分配、尿素氮在氮库中的保存及水稻吸氮状况。结果表明: 施用猪粪显著提高了土壤铵态氮、固定态铵含量和微生物生物量氮,提高了分蘖期尿素氮在各氮库中的贮存,显著增加了水稻产量。与N处理相比,添加抑制剂促进了NH₄⁺的矿物固定和微生物对尿素氮的固持;与NM处理相比,施用抑制剂增加了黏土矿物对¹⁵NH₄⁺的固定。通径分析表明,施用猪粪能促进水稻吸收肥料氮,增加水稻产量;添加抑制剂可通过铵的矿物固定将更多的肥料氮暂时储存;NIM能将更多的氮贮存在微生物生物量氮中,至作物生长后期,铵的释放和微生物周转矿化可为水稻提供更多的有效氮源。在我国北方稻田,NM和NIM处理是推荐的施肥方式。     

The partial replacement of soyabean meal and rapeseed meal with feed grade urea or a slow-release urea and its effect on the performance, metabolism and digestibility in dairy cows

The objectives of the study were to determine the effect of the partial replacement of soyabean meal and rapeseed meal with feed grade urea or a slow-release urea on the performance, metabolism and whole-tract digestibility in mid-lactation dairy cows. Forty-two Holstein–Friesian dairy cows were allocated to one of three dietary treatments in each of three periods of 5 weeks duration in a Latin square design. Control (C) cows were offered a total mixed ration based on grass and maize silages and straight feeds that included 93 g/kg dry matter (DM) soyabean meal and 61 g/kg DM rapeseed meal. Cows that received either of the other two treatments were offered the same basal ration with the replacement of 28 g/kg DM soyabean and 19 g/kg DM rapeseed meal with either 5 g/kg DM feed grade urea (U) or 5.5 g/kg DM of the slow-release urea (S; Optigen^R; Alltech Inc., Kentucky, USA), with the content of maize silage increasing. There was no effect (P > 0.05) of dietary treatment on DM intake, which averaged 22.5 kg/day. Similarly, there was no effect (P > 0.05) of treatment on daily milk or milk fat yield but there was a trend (P = 0.09) for cows offered either of the diets containing urea to have a higher milk fat content (average of 40.1 g/kg for U and S v. 38.9 g/kg for C). Milk true protein concentration and yield were not affected by treatment (P > 0.05). Milk yield from forage and N efficiency (g milk N output/g N intake) were highest (P < 0.01) in cows when offered S and lowest in C, with cows receiving U having intermediate values. Cows offered S also tended to have the highest live weight gain (0.38 kg/day) followed by U (0.23 kg/day) and C (0.01 kg/day; P = 0.07). Plasma urea concentrations were higher (P < 0.05) at 2 and 4 h post feeding in cows when offered U and lowest in C, with animals receiving S having intermediate values. There was no effect (P > 0.05) of treatment on whole-tract digestibility. In conclusion, the partial replacement of soyabean meal and rapeseed meal with feed grade urea or a slow-release urea can be achieved without affecting milk performance or diet digestibility, with the efficiency of conversion of dietary N into milk being improved when the slow-release urea was fed.     

Supplementation with non-fibrous carbohydrates reduced fiber digestibility and did not improve microbial protein synthesis in sheep fed fresh forage of two nutritive values

To determine whether non-fibrous carbohydrate (NFC) supplementation improves fiber digestibility and microbial protein synthesis, 18 Corriedale ewes with a fixed intake level (40 g dry matter (DM)/kg BW0.75) were assigned to three (n = 6) diets: F = 100% fresh temperate forage, FG = 70% forage + 30% barley grain and FGM = 70% forage + 15% barley grain + 15% molasses-based product (MBP, Kalori 3000). Two experimental periods were carried out, with late (P1) and early (P2) vegetative stage forage. For P2, ewes were fitted with ruminal catheters. Forage was distributed at 0900 h, 1300 h, 1800 h and 2300 h, and supplement added at 0900 h and 1800 h meals. Digestibility of the different components of the diets, retained N and rumen microbial protein synthesis were determined. At the end of P2, ruminal pH and N-NH3 concentration were determined hourly for 24 h. Supplementation increased digestibility of DM (P < 0.001) and organic matter (OM; P < 0.001) and reduced NDF digestibility (P = 0.043) in both periods, with greater values in P2 (P = 0.008) for the three diets. Daily mean ruminal pH differed (P < 0.05) among treatments: 6.33 (F), 6.15 (FG) and 6.51 (FGM). The high pH in FGM was attributed to Ca(OH)2 in MBP. Therefore, the decreased fiber digestibility in supplemented diets could not be attributed to pH changes. The mean ruminal concentration of N-NH3 was 18.0 mg/dl, without differences among treatments or sampling hours. Microbial protein synthesis was greater in P2 (8.0 g/day) than in P1 (6.1 g/day; P = 0.006), but treatments did not enhance this parameter. The efficiency of protein synthesis tended to be lower in supplemented groups (16.4, 13.9 and 13.4 in P1, and 20.8, 16.7 and 16.2 g N/kg digestible OM ingested in P2, for F, FG and FGM, respectively; P = 0.07) without differences between supplements. The same tendency was observed for retained N: 2.55, 1.38 and 1.98 in P1, and 2.28, 1.23 and 1.10 g/day in P2, for F, FG and FGM, respectively; P = 0.05). The efficiency of microbial protein synthesis was greater in P2 (P = 0.007). In conclusion, addition of feeds containing NFCs to fresh temperate forage reduced the digestibility of cell walls and did not improve microbial protein synthesis or its efficiency. An increase in these parameters was associated to the early phenological stage of the forage.     

Nitrogen-15 balances for urea and neem coated urea applied to lowland rice following two cowpea cropping systems

Little is known about whether the high N losses from inorganic N fertilizers applied to lowland rice (Oryza sativa L.) are affected by the combined use of either legume green manure or residue with N fertilizers. Field experiments were conducted in 1986 and 1987 on an Andaqueptic Haplaquoll in the Philippines to determine the effect of cowpea [Vigna unguiculata (L.) Walp.] cropping systems before rice on the fate and use efficiency of¹⁵N-labeled, urea and neem cake (Azadirachta indica Juss.) coated urea (NCU) applied to the subsequent transplanted lowland rice crop. The pre-rice cropping systems were fallow, cowpea incorporated at the flowering stage as a green manure, and cowpea grown to maturity with subsequent incorporation of residue remaining after grain and pod removal. The incorporated green manure contained 70 and 67 kg N ha⁻¹ in 1986 and 1987, respectively. The incorporated residue contained 54 and 49 kg N ha⁻¹ in 1986 and 1987, respectively. The unrecovered¹⁵N in the¹⁵N balances for 58 kg N ha⁻¹ applied as urea or NCU ranged from 23 to 34% but was not affected by pre-rice cropping system. The partial pressure of ammoniapNH₃, and floodwater (nitrate + nitrite)-N following application of 29 kg N ha⁻¹ as urea or NCU to 0.05-m-deep floodwater at 14 days after transplanting was not affected by pre-rice cropping system. In plots not fertilized with urea or NCU, green manure contributed an extra 12 and 26 kg N ha⁻¹, to mature rice plants in 1986 and 1987, respectively. The corresponding contributions from residue were 19 and 23 kg N ha⁻¹, respectively. Coating urea with 0.2g neem cake per g urea had no effect on loss of urea-N in either year; however, it significantly increased grain yield (0.4 Mg ha⁻¹) and total plant N (11 kg ha⁻¹) in 1987 but not in 1986.     

Nitrogen supply in cattle coupled with appropriate supply of utilisable crude protein at the duodenum,a precursor to metabolisable protein

The overall objective of this study was to calculate the amount of nitrogen (N) that cattle feed must contain in order to utilise the potential supply of utilisable crude protein at the duodenum provided by their energy intake without incurring a negative N balance, that is, without having to break down body protein. For this purpose, the literature was screened for measurements of net degradation and renal excretion of urea as well as N balances (N intake, faecal N and urinary N) in ruminants (cattle, sheep and goats) fed diets with varying N concentrations. Irreversible loss of N from the body urea pool increased with increasing N intake, but net degradation of urea as a proportion of irreversible loss decreased concurrently. Faecal N appeared not to be influenced by N intake and exceeded 11 g/kg dry matter intake (DMI) only in 7% of the data sets available. Urinary non-urea-N rarely exceeded 4 g/kg DMI and appeared independent of N intake. Urinary urea-N showed a clear dependence of N intake, and it is concluded that 1 g N/kg DMI is sufficient for compensating inevitable N losses in the form of urinary urea. In conclusion, ruminant rations should contain the following N concentrations (per kg DM) to account for obligatory losses: 11 g for compensating losses as faecal N, 4 g for compensating losses as urinary non-urea-N and 1 g for compensating inevitable losses as urinary urea-N. The derived recommendations should be helpful for limiting N excretion where this is desirable for ecological reasons.     

Investigations on the effect of forage source, grinding, and urea supplementation on ruminal fermentation and microbial protein flow in a semi-continuous rumen simulation system

The objective of the present study was to compare the effect of maize silage and grass silage on microbial fermentation and protein flow in a semi-continuous rumen simulation system (Rusitec) when milling screen size (MSS) during grinding was varied. Oven-dried silages were milled through screens of 1, 4 or 9 mm pore size and incubated for 48 h in a Rusitec system. Furthermore, the effect of N supplementation to maize silage (MSS: 4 mm) was investigated and single dose vs. continuous infusion of urea-N were compared. Degradation of organic matter (OM), crude protein (CP), fibre fractions and non-structural carbohydrates (NSC) as well as short-chain fatty acid production differed significantly between forage sources. Urea-N supplementation improved the degradation of NSC, but not that of fibre fractions in maize silage. The way of urea supply had only marginal effects on fermentation characteristics. An increase in MSS, and consequently in mean feed particle size, led to an improvement in the degradation of OM, CP and NSC, but efficiency of microbial net protein synthesis (EMPS; mg microbial N flow/g degraded OM) and the microbial amino acid profile were less affected. EMPS was higher in grass silage than in maize silage and was improved by urea-N supplementation in maize silage. This study indicates that fermentation of NSC as well as EMPS during incubation of maize silage was limited by availability of NH3-N. Furthermore, an increase in MSS above 1 mm seems to improve fermentation of silages in the Rusitec system.     

Effects of a slow-release urea source on absorption of ammonia and endogenous production of urea by cattle

Three experiments were conducted with Angus or Holstein steers to evaluate effects of dietary urea–calcium (a slow rumen-release urea source) on absorption of ammonia N from the gut and urea N production in the liver. Steers were fed a high-grain diet (Experiment 1) or an all-forage diet (Experiments 2 and 3). Urea or urea–calcium (0.25 g/kg body weight) was dosed into the esophagus (Experiments 1 and 2) or rumen (Experiment 3), and blood samples were serially collected for 180 min. Blood concentrations of ammonia N and urea N were measured in all experiments, and net flux of metabolites across splanchnic tissues was measured in Experiment 3. Compared to urea, urea–calcium reduced (P<0.05) plasma concentrations of ammonia N in steers fed all-forage diets, and tended (P<0.06) to reduce arterial glucose concentrations in Experiment 3. Plasma concentrations of urea N were not affected by treatment in any experiment. Treatment and time post-dosing interactions (P<0.05) in Experiment 3 were due to increased ruminal fluid concentrations of ammonia N, net release of ammonia N by portal-drained viscera and total splanchnic tissues with urea versus urea–calcium treatment shortly after dosing. Similar interactions (P<0.05) indicated that urea caused higher hepatic glucose release and increased l-lactate release by total splanchnic tissues after dosing than urea–calcium. Urea–calcium was effective in mitigating rapid ammonia release in the rumen and subsequent effects on glucose and lactate metabolism.     

Incorporation of N from intravenously administered 15N labelled urea into the bacterial protein in the sheep.

The experiment carried out on two wethers demonstrated that nitrogen of intravenously injected urea, labelled with 15N was incorporated into total and bacterial nitrogen fraction of the digesta flowing through the rumen and duodenum. The amount of 15N in the bacterial fraction flowing throught the rumen and duodenum was relatively low in comparison with the amount of 15N in the total nitrogen (14,8% and 8,1% in the rumen and 6,6% and 7,9% in the duodenum. The ratio of the amount of bacterial-N to total-N in the rumen content (12,7 and 7,5%) was only slightly lower than the ratio of bacterial 15N to total 15N. In the duodenum this ratio was a little higher (8,7 and 10,0%). Blood urea nitrogen was utilized only partly in biosynthesis of bacterial protein. The results showed that only a small amount of blood urea nitrogen retained in the organism was utilized for microbial protein synthesis and the majority in some different way.     

Intake and digestion by lambs of dwarf elephant grass (Pennisetum purpureum Schum. cv. Mott) hay or hay supplemented with urea and different levels of cracked corn grain

This study was carried out to evaluate intake, digestibility, ruminal fermentation, nitrogen (N) retention and ruminal microbial protein synthesis in lambs fed dwarf elephant grass (Pennisetum purpureum Schum. cv. Mott) hay or hay supplemented with urea and 0, 5, 10 or 15 g/kg of live weight (LW) of cracked corn grain. Ten lambs (mean LW of 28 ± 0.9 kg), housed in metabolic cages, were used in a double 5 × 5 Latin Square experiment. Except fibre intake and digestibility, which was higher, the intake and digestibility of the others feed components, as well as ruminal microbial protein synthesis and N retention were lower in non-supplemented lambs. Corn supplementation increased total dry matter (DM) (P<0.05), organic matter (OM), non-structural carbohydrate (NSC) and energy intake (P<0.01) but decreased total neutral detergent fibre (aNDFom) (P<0.01) intake, as well as OM and aNDFom intake from the hay (P<0.01). Apparent DM, OM and energy digestibility, as well as OM true digestibility (OMTD) increased (P<0.01), and aNDFom digestibility decreased linearly (P<0.01) as corn supplementation increased. Total N intake was not influenced but, apparent and true N digestibility, as well as urinary N excretion decreased (P<0.01), and ruminal microbial N entering the small intestine increased linearly (P<0.01) as corn supplementation increased. However, the efficiency of ruminal microbial protein synthesis was similar for all treatments. Mean ruminal pH values and ammonia N concentrations decreased linearly (P<0.01) with level of corn supplementation. Ammonia N and amino acid, as well as peptide concentrations in ruminal fluid were quadratically related (P<0.01) with the time after feeding. Corn supplementation had a linear additive effect on total dry matter and digestible energy intake, as well as on N retention, but a linear negative effect on hay intake and on fibre digestibility. However, decreased forage digestibility by animals was probably neither related to lower ruminal pH, which values were always higher than 7.0, nor related to ruminal sugar concentrations, which were similar for all treatments.     

High-concentrate diets based on forages harvested at different maturity stages affect ruminal synthesis of B vitamins in lactating dairy cows

Effects of plant maturity on apparent ruminal synthesis and post-ruminal supply of B vitamins were evaluated in two feeding trials. Diets containing alfalfa (Trial 1) or orchardgrass (Trial 2) silages harvested either (1) early cut, less mature (EC) or (2) late cut, more mature (LC) as the sole forage were offered to ruminally and duodenally cannulated lactating Holstein cows in crossover design experiments. In Trial 1, conducted with 16 cows (569±43 kg of empty BW (ruminal content removed) and 43.7±8.6 kg/day of 3.5% fat-corrected milk yield; mean±SD) in two 17-day treatment periods, both diets provided ~22% forage NDF and 27% total NDF, and the forage-to-concentrate ratios were 53 : 47 and 42 : 58 for EC and LC, respectively. In Trial 2, conducted with 13 cows (588±55 kg of empty BW and 43.7±7.7 kg/day of 3.5% fat-corrected milk yield; mean±SD) in two 18-day treatment periods, both diets provided ~25% forage NDF and 31% total NDF; the forage-to-concentrate ratios were 58 : 42 and 46 : 54 for EC and LC, respectively. Thiamin, riboflavin, niacin, vitamin B₆, folates and vitamin B₁₂ were measured in feed and duodenal content. Apparent ruminal synthesis was calculated as the duodenal flow minus the intake. Diets based on EC alfalfa decreased the amounts of thiamin, niacin and folates reaching the duodenum, whereas diets based on EC orchardgrass increased riboflavin duodenal flow. Daily apparent ruminal synthesis of thiamin, riboflavin, niacin and vitamin B₆ were correlated negatively with their intake, suggesting a microbial regulation of their concentration in the rumen. Vitamin B₁₂ apparent ruminal synthesis was correlated negatively with total volatile fatty acids concentration, but positively with ruminal pH and microbial N duodenal flow.     

Transfer of blood urea nitrogen to cecal microbial nitrogen is increased by mannitol feeding in growing rabbits fed timothy hay diet

The presence of the fermentable sugar d-mannitol in the diet improves nitrogen (N) utilization in rabbits. To clarify the mechanism by which d-mannitol improves N utilization, we studied the effect of d-mannitol on the fate of blood urea N in growing rabbits. Growing rabbits received a control diet or a diet containing d-mannitol, which were formulated by adding 80 g/kg glucose or d-mannitol to timothy hay. After 9 days of feeding of the experimental diets, ¹⁵N-urea was administrated intravenously under anesthesia 1 h before slaughter. The blood urea level (concentration of both urea N (43.6% of the control group (CG), P < 0.05) and ¹⁵N (95% of the CG, P < 0.05) in blood serum) was reduced in the mannitol group. The concentration and amount of N, and ¹⁵N atom % excess in the contents of the cecum and colon were higher (P < 0.05) in the rabbits fed the mannitol diet than in rabbits fed the control diet, especially in the cecum. The consumption of mannitol caused bacterial proliferation in the cecum characterized by marked short-chain fatty acid production (165% of the CG, P < 0.05), decreased cecal ammonia N (73% of the CG, P < 0.05) and elevated cecal bacterial N (150% of the CG, P < 0.05). On the other hand, addition of d-mannitol to the diet decreased N (80% of the CG, P < 0.05) and ¹⁵N (77% of the CG, P < 0.05) excretion in the urine. These results indicate that d-mannitol increases the transfer of blood urea N to the large intestine, where it is used for bacterial N synthesis.     

Ruminal degradability of 15N labelled ribonucleic acid in grass

The ruminal degradation of RNA in rye grass (Lolium perenne) was studied using the bag method. A non-lactating cow (BW 550 kg) fitted with a rumen cannula was used and fed twice daily at maintenance level with a chopped grass hay-based ration containing 30% ground barley. Rye grass, labelled during growth by fertilization with 15N2-urea (9.5 atom% 15N, 20 g N/m2), was cut at seven stages of growth and maturity and freeze-dried. RNA-N represented 6 to 17% of total N. Labelled grass samples (milled to 5.0 mm screen, 5.0+/-0.1 g DM) were incubated in polyester bags (100 x 200 mm, pore size 50 microm) in the rumen for periods of 1, 3, 6, 9, 12, 24, and 48 h. Data of N and RNA disappearances from the bags were fitted to an exponential equation to estimate parameters of degradation. The effective degradability of RNA in the rumen averaged 90+/-4%, for N it was 11% units lower (P < 0.001). Degradability of RNA was correlated to that of N (R2 = 0.92). Degradability of RNA (R2 = 0.96) and N (R2 = 0.93) decreased with increasing fibre content of grass. Increasing the fibre content by 1% diminished the degradability of RNA and N by 1.1% units and 2.4% units, respectively (P < 0.001). Assuming a microbial protein synthesis in the rumen of 150 g/kg DOM, a N: RNA ratio of 1:1.35 in rumen microbes and a rumen outflow rate of 0.06 h(-1), a model calculation indicates that about 9 to 19% of duodenal RNA are of dietary origin in animals fed grass. This should be taken into account for the calculation of microbial N on the basis of RNA as marker.     

Effects of linseed lipids fed as rolled seeds, extruded seeds or oil on organic matter and crude protein digestion in cows

Effects of fatty acids of linseed in different forms, on ruminal fermentation and digestibility were studied in dry cows fitted with ruminal and duodenal cannulas. Four diets based on maize silage, lucerne hay and concentrates (65/10/25 dry matter (DM)) were compared in a 4 × 4 Latin square design experiment where the diets were: control diet (C), diet RL supplied 75 g/kg DM rolled linseeds, diet EL supplied 75 g/kg DM extruded linseeds, and diet LO supplied 26 g/kg DM linseed oil and 49 g/kg DM linseed meal. The diets did not differ in total organic matter (OM) and fibre digestibility, in forestomach and intestinal OM digestibility, and in duodenal N flow. Microbial N duodenal flow tended to be lower for RL versus C diet (P<0.1). Extrusion did not reduce ruminal crude protein (CP) degradation in vivo and in situ. Volatile fatty acid concentration and pattern, and protozoa concentration in the rumen, did not vary among diets. Results confirm the absence of a negative effect of a moderate supply of linseed on rumen function, as well as no effect of extrusion on its ruminal CP degradability.