Methanol-fermentation protein in veal calf nutrition

In an experiment with 4 × 12 male veal calves, the influence of 0, 2.5, 5.0 and 7.5% ICI protein and sweet whey powder in a milk replacer on growth; feed efficiency; blood concentrations of urea, uric acid and allantoin; and on the iron status, colour of the meat and carcass grade of the animals was investigated. The proportion of skimmed milk powder in the milk replacer was thus reduced in protein equivalence by 5, 10 and 15% (groups II–IV).Feeding with 2.5, 5 and 7.5% ICI protein in the milk replacer gave, over the whole fattening period (60–150 kg), about the same rates of growth and feed efficiency as the exclusive use of milk protein. While the urea content in the serum was not significantly influenced by the protein sources used, the uric acid and allantoin levels of the groups given ICI protein had risen greatly at 5 weeks and at the end of fattening experiment. Despite the higher total iron supply to groups II, III and IV, the criteria indicating iron supply such as haemoglobin, haematocrit and myoglobin levels did not show any increase compared to group I. Similarly, no significant differences in meat colour and carcass grade were found among the experimental groups.For veal calves the level of ICI protein in the milk replacer can be up to 7.5% during the entire feeding period.     

Effects of level and source of supplemental protein in a concentrate-based diet on growth performance of Boer × Spanish wether goats

Boer×Spanish wether goats (50; 26.8±4.04 kg initial BW and 7 months of age) were used in a 27 week experiment to determine effects of two levels of CP (13 and 19%) and five protein sources varying in ruminal degradability (blood, corn gluten, feather, fish and soybean meals). Diets were 70% concentrate, had a ratio of ruminally degraded intake protein (DIP) to total digestible nutrients (TDNs) of at least 0.09 and were formulated to maximize ruminally undegradaed protein from supplemental protein sources. There were no interactions between dietary CP level and supplemental protein source. Voluntary DM intake (1043, 1089, 1153, 1086 and 1112 g per day; S.E.=74.4), ADG (136, 134, 143, 145 and 138 g per day; S.E.=9.8) and gain efficiency (ADG:DM intake) (131, 124, 125, 136 and 128 g/kg for blood, corn gluten, feather, fish and soybean meals, respectively; S.E.=5.7) were similar between CP levels and among sources of supplemental protein. In summary, with a dietary concentrate level of 70% and at least 13% CP, differences in amino acid profiles among blood, corn gluten, feather, fish and soybean meals did not impact rate or efficiency of growth by Boer×Spanish wethers.     

Induction and inhibition of the allantoin permease in Saccharomyces cerevisiae.

Allantoin uptake in Saccharomyces cerevisiae is mediated by an energy-dependent, low-Km, active transport system. However, there is at present little information concerning its regulation. In view of this, we investigated the control of alloantoin transport and found that it was regulated quite differently from the other pathway components. Preincubation of appropriate mutant cultures with purified allantoate (commercial preparations contain 17% allantoin), urea, or oxalurate did not significantly increase allantoin uptake. Preincubation with allantoin, however, resulted in a 10- to 15-fold increase in the rate of allantoin accumulation. Two allantoin analogs were also found to elicit dramatic increases in allantoin uptake. Hydantoin and hydantoin acetic acid were able to induce allantoin transport to 63 and 95% of the levels observed with allantoin. Neither of these compounds was able to serve as a sole nitrogen source for S. cerevisiae, and they may be non-metabolizable inducers of the allantoin permease. The rna1 gene product appeared to be required for allantoin permease induction, suggesting that control was exerted at the level of gene expression. In addition, we have shown that allantoin uptake is not unidirectional; efflux merely occurs at a very low rate. Allantoin uptake is also transinhibited by addition of certain amino acids to the culture medium, and several models concerning the operation of such inhibition were discussed.     

The Surveys on the Urea and Allantoin Excretion and Other Criteria from Rats Fed “Threonine Imbalanced” or “Corrected” Diet

Weanling male rats of Wistar strain were fed ad libitum 7 or 8% casein-a-starch diet supplemented with 0.3% dl-methionine with ( + Thr) or without ( ?Thr) added 0.36% dl-threonine for 12 days. Urine was collected 3 times for 3 days each at one-day interval. The urine sample was analysed for allantoin, urea and total N.

There was no difference between the values of growth rate, liver and carcass fat content, protein efficiency ratio and % N retention of ?Thr and +Thr groups, whereas urea excretions of ?Thr groups were significantly higher (over 2 times) than those of +Thr and allantoin excretions were consistently lower in the former when dietary protein level was 8%.     

Urease Is Not Essential for Ureide Degradation in Soybean

The hypothesis that soybean (Glycine max L. [Merrill]) catabolizes ureides to urea to a physiologically significant extent was tested and rejected. Urease-negative (eu3-e1/eu3-e1) plants were supported by fixed N2 or by 2 mM NH4NO3, so that xylem-borne nitrogen contained predominantly ureides (allantoin and allantoic acid) or amide amino acids, respectively. Seed nitrogen yield was equal on either nitrogen regime, although 35-d-old fixing plants accumulated about 6 times more leaf urea. In callus, lack of an active urease reduced growth on either arginine or allantoin as the sole nitrogen source, but the reduction was greater on arginine (73%) than on allantoin (39%). Furthermore, urease-negative cells accumulated 17 times more urea than urease-positive cells on arginine; for allantoin the ratio was 1.8. Urease-negative callus accumulated urea at 3% the rate of seedlings. To test whether urea accumulating in urease-negative seedlings was derived from ureides, seeds were first allowed to imbibe in 1 mM allopurinol, an inhibitor of ureide formation. Seedling ureides were decreased by 90%, but urea levels were unchanged. Thus, ureides are poor precursors of urea, which was confirmed in seedlings that converted no more than 5% of seed-absorbed [14C-ureido]allantoate to [14C]urea, whereas 40 to 70% of [14C-guanido]arginine was recovered as [14C]urea.     

Effect of protein intake on amino acid catabolism and gluconeogenesis by isolated hepatocytes from the cat (Felis domestica)

Cats were fed 17.5% (LP) and 70% (HP) diets and hepatocytes were prepared from them. Rates of gluconeogenesis from pyruvate, alanine and threonine (10 mM) were unaffected by protein intake but 10 mM glutamine was converted faster by cells from HP fed animals. Rates of oxidation of alanine, threonine and glutamine and flux rates of tyrosine aminotransferase and tryptophan 2,3-dioxygenase were greater in cells from HP fed cats at all amino acid concentrations used. Proteolysis was indicated by urea production which was higher in cells from HP fed cats but was reduced significantly by leupeptin.     

Bacterial protein meal in diets for pigs and minks: Comparative studies on protein turnover rate and urinary excretion of purine base derivatives

Abstract

The effect of increasing the dietary content of bacterial protein meal (BPM) on protein turnover rate, and on nucleic acid and creatinine metabolism in growing minks and pigs was investigated in two experiments. In each experiment, 16 animals were allocated to four experimental diets. The diets containing no BPM served as controls, i.e. for minks diet M1, for pigs P1; the experimental diets contained increasing levels of BPM to replace fish meal (minks) or soybean meal (pigs), so that up to 17% (P2), 20% (M2), 35% (P3), 40% (M3), 52% (P4), and 60% (M4) of digestible N was BPM derived. Protein turnover rate was measured by means of the end-product method using [¹⁵N]glycine as tracer and urinary nitrogen as end-product. In minks, protein flux, synthesis, and breakdown increased significantly with increasing dietary BPM. In pigs, diet had no observed effect on protein turnover rate. The intake of nucleic acid nitrogen (NAN) increased from 0.15 g/kg W^0.75 on M1 to 0.26 g/kg W^0.75 on M3 and M4 in the mink experiment, and from 0.08 g/kg W^0.75 on P1 to 0.33 g/kg W^0.75 on P4 in the pig experiment. Increased NAN intake led, in both experiments, to increased allantoin excretion. Analysis of species effects showed that minks excreted 1.72 mmol/kg W^0.75 of allantoin, significantly more than the 0.95 mmol/kg W^0.75 excreted by pigs. In minks, approximately 96% of the excreted purine base derivatives consisted of allantoin, whereas in pigs approximately 93% did. Thus, increasing the dietary content of BPM increased protein turnover rate in minks but not in pigs, and allantoin excretion increased with increasing dietary BPM although it seemed that mink decomposed purine bases to their end-product more completely than pigs did. Collectively these data show that BPM is a suitable protein source for pigs and mink, and recorded differences between species were to a large extent due to differences in protein retention capacity and muscle mass.     

The role of energy,serine, glycine,and 1-carbon units in the cost of nitrogen excretion in mammals and birds

The efficiency with which dietary protein is used affects the nitrogen excretion by the animal and the environmental impact of animal production. Urea and uric acid are the main nitrogen excretion products resulting from amino acid catabolism in mammals and birds, respectively. Nitrogen excretion can be reduced by using low-protein diets supplemented with free amino acids to ensure that essential amino acids are not limiting performance. However, there are questions whether the capacity to synthesize certain nonessential amino acids is sufficient when low-protein diets are used. This includes glycine, which is used for uric acid synthesis. Nitrogen excretion not only implies a nitrogen and energy loss in the urine, but energy is also required to synthesize the excretion products. The objective of this study was to quantify the energy and metabolic requirements for nitrogen excretion products in the urine. The stoichiometry of reactions to synthesize urea, uric acid, allantoin, and creatinine was established using information from a publicly available database. The energy cost was at least 40.3, 60.7, 64.7, and 65.4 kJ/g excreted N for urea, uric acid, allantoin, and creatinine, respectively, of which 56, 56, 47, and 85% were retained in the excretion product. Data from a broiler study were used to carry out a flux balance analysis for nitrogen, serine, glycine, and so-called 1-carbon units. The flux balance indicated that the glycine intake was insufficient to cover the requirements for growth and uric acid excretion. The serine intake was also insufficient to cover the glycine deficiency, underlining the importance of the de novo synthesis of serine and glycine. One-carbon units are also a component of uric acid and can be synthesized from serine and glycine. There are indications that the de novo synthesis of 1-carbon units may be a “weak link” in metabolism, because of the stoichiometric dependency between the synthesized 1-carbon units and glycine. The capacity to catabolize excess 1-carbon units may be limited, especially in birds fed low-protein diets. Therefore, there may be an upper limit to the 1-carbon-to-glycine requirement ratio in relation to nutrients that supply 1-carbon units and glycine. The ratio can be reduced by increasing uric acid excretion (i.e., reducing protein deposition) or by dietary supplementation with glycine. The hypothesis that the 1-carbon-to-glycine requirement ratio should be lower than the supply ratio provides a plausible explanation for the growth reduction in low-protein diets and the positive response to the dietary glycine supply.     

Effect of Adding Branched-chain Amino Acids to a Nicotinic Acid-free,Low-protein Diet on the Conversion Ratio of Tryptophan to Nicotinamide in Rats

The effects of adding branched-chain amino acids to a nicotinic acid-free, low-protein diet on the conversion of tryptophan to nicotinamide were investigated in rats. The conversion ratio [urinary excretion of nicotinamide and its metabolites (μmol/day) × 100/tryptophan intake during urine collection (μmol/day)] was significantly lower in the groups fed with the 3% Leu-, Val-, or Ile-added diet than in the group fed with the control diet. Namely, the inhibition of this conversion was observed not only by the addition of Leu, but also by the addition of Val or Ile. The addition of Ile and/or Val to the Leu-added diet did not antagonize the Leu effect.     

The effect of roughage to concentrate ratio in the diet on nitrogen and purine metabolism in dairy cows

Four lactating dairy cows were used in two experiments to study the effects of the roughage to concentrate ratio in the diet on nitrogen balance, plasma urea, urinary urea, milk urea and urinary purine derivatives. The use of the allantoin to creatinine ratio in spot samples of urine as an index of the urinary allantoin excretion was also evaluated. Four isoenergetic and isonitrogenous diets were formulated according to a 2 × 2 factorial arrangement. Factor I was concentrate content. The roughage to concentrate ratios were 65:35 and 35:65 for the high roughage and high concentrate diets, respectively. Factor II was fat content, which was 2.8% and 5.8% for the low and high fat diets, respectively. In Experiment 1 cows were fed diets with low fat content, and in Experiment 2 cows were fed diets with high fat content. In both experiments, diets were fed according to a change-over design. Nitrogen balance was not affected by the treatments. In cows fed high concentrate diets the amount and the proportion of nitrogen excreted in milk, as well as milk production was higher than in cows fed the high roughage diets. In both experiments, as an overall effect, the urea levels in plasma, urine and morning milk were higher, although the total urinary excretion of urea was lower, for the high concentrate diets. Urinary allantoin excretion was higher, although not significantly in Experiment 1, for the high concentrate diets. The allantoin to creatinine ratio in spot samples of urine showed the same pattern as the total allantoin excretion. Urinary creatinine excretion appeared to be affected by the diet.     

Further Studies on the Effect of Amino Acid Supplementation on the Urinary Nitrogen Compounds in Rats Fed a Low-casein Diet

When 8% casein basal diet was supplemented with 0.3% dl-methionine or 0.3% dl-methionine plus 0.36% dl- or 0.18% l-threonine, the changes in urinary excretions of urea and allantoin were examined in weanling male rats of Wistar strain with the observations on the body weight gain and % nitrogen retention. Carbohydrate sources used were sucrose or an equimolar mixture of glucose and fructose (G-F) in place of pregelatinized starch used in the previous experiments.

In contrast to the previous results, differences in nitrogen utilization, expressed in term of growth rate or % nitrogen retention, became significant by the addition of 0.3% methionine to the basal diet and it was further increased by the simultaneous supplementation with 0.36% dl- or 0.18% l-threonine.

Urea excretion was the main variable in total urinary nitrogen output to cause the significant difference in % nitrogen retention between the groups. As postulated in the previous paper, thus, the use of sucrose or G-F mixture considerably exaggerated these group-differences in such various indices as body weight gain and % nitrogen retention, and this trend became more distinct in the urea and allantoin excretions.

Liver arginase activity inversely changed with urea excretion, but proportionately to the qualitative improvement of dietary protein by the addition of methionine or methionine plus threonine. Changes in liver glutamic dehydrogenase activity were also parallel with the improvement of dietary protein quality.     

Tolerable amounts of amino acids for human supplementation: summary and lessons from published peer-reviewed studies

Amino acid supplementation may be indicated to correct for insufficient amino acid intake in healthy individuals, and in specific physiological or pathophysiological situations. However, there is a concern to not supplement beyond the tolerable upper intake level (UL) by determining parameters of no-observed-adverse-effect level (NOAEL) or lowest-observed-adverse-effect level (LOAEL) for each amino acid. Since the NOAEL and LOAEL values are at least one order of magnitude different when comparing the values obtained in rats and humans, the aim of this review is to evaluate to what extent the amino acid UL measured in the rat model, when referenced to the dietary usual consumption (UC) and dietary requirement (RQ) for indispensable amino acids, may be used as an approximation of the UL in humans. This review then compares the ratios of the NOAEL or LOAEL over UC and RQ in the rat model with the same ratios calculated in humans for the nine amino acids (arginine, serine, glycine, histidine, leucine, lysine, methionine, phenylalanine, and tryptophan) for which this comparison can be done. From the calculations made, it appears that for these 9 amino acids, the calculated ratios for rats and humans, although rather different for several amino acids, remains for all of them in the same order of magnitude. For tryptophan, tyrosine, and valine, the ratios calculated in rats are markedly different according to the sex of animals, raising the view that it may be also the case in humans.

     

Biochemical Studies on Rice Bran Lipase

Some chemical properties of the rice bran lipase were studied. The enzyme protein contained 14.98% nitrogen and consisted of 312 amino acid residues. It also contained a certain amount of lipid. The amino-terminal amino acids of the enzyme protein were shown to be glutamic acid and the carboxyl-terminal amino acids to be glycine and serine. The treatment of the enzyme protein with 8 m urea containing 1×10^?3m EDTA (ethyl-enediaminetetraacetic acid) seemed to cause dissociation of the subunits of the enzyme protein. From this observation and the results of the terminal amino acids analysis, it was presumed that the enzyme protein was composed of at least two types of subunits.     

Water and nonelectrolyte permeability of plant cell membranes after short term application of amino acids and phosphorylated amino acids

The effects of amino acids (aa) and N-(diisopropyloxyphosphoryl)-amino acids (DIPP-aa) on cell membranes were investigated by evaluating water and methyl urea permeability. Permeability coefficients P_f and P_s were determined by standard osmotic methods for cells ofPisum sativum stem base epidermis after 20 min exposure to a 5 mM solution of each aa and DIPP-aa. The P_f value ofP. sativum epidermal cells (untreated controls) was 1.3 ± 0.4 × 10^-3 μm s^-1. Treat ments with the diisopropyl-oxyphosphoryl derivatives of three one charged and three polar amino acids (serine, threonine, asparagine, and aspartic acid) and unsubstituted (free) serine and threonine increased water permeability up to about two fold of the control value. Serine and threonine and their DIPP-derivatives increased methyl urea permeability (controls 1.03 ± 0.09 × 10^-3 μm s^-1) 30 to 80 percent Other amino acids and their DIPP-derivatives caused small or insignificant changes of water permeability. Only certain polar amino acids and their DIPP-derivatives increased the osmotic water and methyl urea permeation through the plasma membrane. The specificity of these molecules on plasma membranes suggests that the active amino acids (serine and threonine) and their DIPP-derivatives interact with charged membrane molecules. The relatively small changes in water and methyl urea permeability may indicate that the effective aa’s and their DIPP-derivatives interact with phospholipids rather than aquaporin. A concurring alteration of water channel proteins, however, cannot excluded.     

Recovery of tryptophan from 25-minute acid hydrolysates of protein

It was found that thioglycolic acid prevents destruction of tryptophan during rapid hydrolysis of protein with a trifluoroacetic acid/HCl mixture (1:2, v/v) at 166 degrees C for 25 or 50 min. The addition of 5% (v/v) thioglycolic acid gave the maximum tryptophan recovery (88.3%) for a 25-min hydrolysate of lysozyme. Tryptophan recoveries varied slightly among three different proteins; 88% for lysozyme, 73% for alpha-chymotrypsinogen A, and 85% for apomyoglobin. However, when extrapolated to zero time, the values were close to one another: 94, 87, and 88%, respectively. The addition of thioglycolic acid was also advantageous for recovering amino acids other than tryptophan. Particularly, yields of carboxymethylcysteine and methionine were greatly improved. This modified rapid hydrolysis method gave satisfactory results without the need for separate analyses of tryptophan and cysteine, provided proteins were reduced and carboxymethylated prior to hydrolysis.     

Metabolism and excretion of nitrogen during metamorphosis and egg production in the sawfly, Neodiprion sertifer

Measurements of whole-body dry matter, total nitrogen, water-soluble protein, amino acids, and uric acid were determined at successive stages during metamorphosis in Neodiprion sertifer. The major change was in the uric acid fraction: in females, it increased up to the non-pharate pupa and then decreased during the subsequent stages of adult development and egg production; in males, it continued to increase during adult development. The decline of uric acid could not be explained by the accumulation of allantoin, allantoic acid, urea, or uric acid riboside. Examination of amino acid levels in the gut revealed an accumulation in the pupa followed by a depletion at the onset of adult development. This was followed by an excretory phase marked by the progressive accumulation of large quantities of uric acid and small quantities of urea, ammonia, and amino acids during the formation of the meconium. Amino acid analysis of the meconium revealed the presence of large proportions of proline, hydroxyproline, and histidine in comparison with the other amino acids.     

Dietary Protein Quality and Liver Arginase Activity of Rats

The arginase activity of the liver of rats was measured after they had been maintained for 11 or 12 days on diets containing natural proteins (casein or gluten) or amino acid mixtures of various tryptophan levels.

Specific activity or total arginase activity increased with the increasing protein quality. Liver arginase activity of the casein group was significantly higher than that of the gluten group in every case for 27-, 61-, and 158-day-old rats. In the case of amino acid diets, the arginase increased with the increments of tryptophan levels up to the “tentative” minimum requirement in the diet. Moreover, these alterations in arginase activity varied inversely with the urinary urea excretion.

From the results, it was assumed that the total activity of liver arginase is not necessarily determined only by the metabolic needs for urea biosynthesis.     

Influence of amino acid availability on protein turnover in perfused skeletal muscle

The effects of amino acids on protein turnover in skeletal muscle were determined in the perfused rat hemicorpus preparation. Perfusion of preparations from fasted young rats (81±2 g) with medium containing either a complete mixture of amino acids at five times (5×) their normal plasma levels, a mixture of leucine, isoleucine, and valine at 5× or 10× levels, or leucine alone (10×) resulted in a 25–50% increase in muscle protein synthesis and a 30% decrease in protein degradation compared to fasted controls perfused in the absence of exogenously added amino acids. When the branched-chain amino acids were omitted from the complete mixture, the remaining amino acids (5×) had no effect on protein turnover. The complete mixture at 1× levels was also ineffective. Comparison of the effects of amino acids with those of glucose and palmitate indicated that amino acids were not acting by providing substrates for energy metabolism. The stimulatory effect of amino acids on protein synthesis was associated with a facilitated rate of peptide-chain initiation as evidenced by a relative decrease in the level of ribosomal subunits. This response was not as great as that produced by insulin, and the amino acids did not augment the effect of insulin. Although protein synthesis in preparations from fed young rats (130±3 g) was stimulated by the addition of a mixture of the branched-chain amino acids (5×) to about the same extent as that observed in the fasted young rats, protein degradation was not affected. Furthermore, neither synthesis nor degradation were affected in preparations from fasted older rats (203±9 g) suggesting that the age and or nitritional state of the animal may influence the response of skeletal muscle to altered amino acid levels.     

Bacterial protein meal in diets for pigs and minks: comparative studies on protein turnover rate and urinary excretion of purine base derivatives

The effect of increasing the dietary content of bacterial protein meal (BPM) on protein turnover rate, and on nucleic acid and creatinine metabolism in growing minks and pigs was investigated in two experiments. In each experiment, 16 animals were allocated to four experimental diets. The diets containing no BPM served as controls, i.e. for minks diet M1, for pigs P1; the experimental diets contained increasing levels of BPM to replace fish meal (minks) or soybean meal (pigs), so that up to 17% (P2), 20% (M2), 35% (P3), 40% (M3), 52% (P4), and 60% (M4) of digestible N was BPM derived. Protein turnover rate was measured by means of the end-product method using [15N]glycine as tracer and urinary nitrogen as end-product. In minks, protein flux, synthesis, and breakdown increased significantly with increasing dietary BPM. In pigs, diet had no observed effect on protein turnover rate. The intake of nucleic acid nitrogen (NAN) increased from 0.15 g/kg W0.75 on M1 to 0.26 g/kg W0.75 on M3 and M4 in the mink experiment, and from 0.08 g/kg W0.75 on P1 to 0.33 g/kg W0.75 on P4 in the pig experiment. Increased NAN intake led, in both experiments, to increased allantoin excretion. Analysis of species effects showed that minks excreted 1.72 mmol/ kg W0.75 of allantoin, significantly more than the 0.95 mmol/kg W0.75 excreted by pigs. In minks, approximately 96% of the excreted purine base derivatives consisted of allantoin, whereas in pigs approximately 93% did. Thus, increasing the dietary content of BPM increased protein turnover rate in minks but not in pigs, and allantoin excretion increased with increasing dietary BPM although it seemed that mink decomposed purine bases to their end-product more completely than pigs did. Collectively these data show that BPM is a suitable protein source for pigs and mink, and recorded differences between species were to a large extent due to differences in protein retention capacity and muscle mass.