Contribution of dietary arginine to nitrogen utilisation and excretion in juvenile sea bass (Dicentrarchus labrax) fed diets differing in protein source

The role of dietary arginine in affecting nitrogen utilisation and excretion was studied in juvenile European sea bass (Dicentrarchus labrax) fed for 72 days with diets differing in protein sources (plant protein-based (PM) and fish-meal-based (FM)). Fish growth performance and nitrogen utilisation revealed that dietary Arg surplus was beneficial only in PM diets. Dietary Arg level significantly affected postprandial plasma urea concentrations. Hepatic arginase activity increased (P<0.05) in response to dietary Arg surplus in fish fed plant protein diets; conversely ornithine transcarbamylase activity was very low and inversely related to arginine intake. No hepatic carbamoyl phosphate synthetase III activity was detected. Dietary arginine levels did not affect glutamate dehydrogenase activity. A strong linear relationship was found between liver arginase activity and daily urea-N excretion. Dietary Arg excess reduced the proportion of total ammonia nitrogen excreted and increased the contribution of urea-N over the total N excretion irrespective of dietary protein source. Plasma and excretion data combined with enzyme activities suggest that dietary Arg degradation via hepatic arginase is a major pathway for ureagenesis and that ornithine-urea cycle is not completely functional in juvenile sea bass liver.     

Effect of arginine on the arginase activity and urea content of the brain and liver of rats acclimating to cold

At cold stress (3 days exposition at 2--4 degrees C) the urea formation in rats brain and liver does not become more active, the content of extraerythrocytic hemoglobin and the total peroxydase activity increase in blood serum, the animals sensitivity to the action of hyperbaric oxygenation (HB0) grows. At cold adaptation (45 days at 2--4 degrees C) the urea content in tissues and the activity of arginase in liver increase, the concentration of extraerythrocytic hemoglobin and the total peroxydase activity normalize, animals become more resistant to HB0. Every day administration of arginine during 3-day cold effect makes the brain and liver arginase on 42 and 28% more active, increases the urea content on 26 and 19%, stabilizes the erythrocytic membranes. The animals protected by arginine against cold are more resistant to the action of HB0.     

An Inverse Relationship between Liver Arginase Activity and Urea Excretion in Rats

Liver arginase activity in rats fed graded levels of diammonium citrate in high and low casein diets, was measured with simultaneous determination of urea excretion. The arginase activity changed inversely with the urea excretion.

Moreover, when the amino acid balance was quantitatively changed by varying threonine level alone, a similar relationship between total liver arginase activity and urea excretion was again observed.

From these results, the early teleological assumptions are most unlikely in that liver arginase activity increases with the decrease in dietary protein quality and that the change in activity can be used as an index of dietary protein quality.     

Peculiarities of urea distribution in tissues of rats of different age

The content of urea was studied in protein-free filtrates of the liver kidneys, skeletal muscles, myocardium, spleen, brain tissues and blood serum as well as in urine of 1, 3, 6, 12 and 24-month rats. It is shown that at the age of 6 months the content of urine in most tissues under study is significantly decreased (by 42-73%), at the age of 12 months in the spleen and at the age of 24 months in the brain tissues as compared to the one-month animals. The level of urine decrease in the liver and brain tissues of 24-month animals is less pronounced than in other tissues, that corresponds to age peculiarities of their protein metabolism. A decrease of blood consumption per weight unit and a relative increase in the amount of nitrogen excreted with urine are observed with ageing. The arginase activity in the liver decreases essentially only in 3-month animals. A conclusion is drawn that peculiarities of food consumption and the character of changes in the urea content in tissues and urine are adaptation manifestation of an age decrease in the intensity of nitrogen metabolism and protein demand of the organism.     

Interaction Between Nickel and Protein Source in the Ruminant

Eighty growing steers were used to determine the effect of nickel supplementation on performance and metabolic parameters of steers fed corn silage-based diets supplemented with different crude protein sources. Crude protein sources examined included: (1) soybean meal, (2) blood meal, (3) urea, and (4) blood meal-urea (two-thirds of supplemental nitrogen from blood meal and one-third from urea). The protein sources differed in ruminal degradability, nitrogen solubility, and nickel content. Nickel was added within each protein treatment to supply either 0 or 5 ppm of supplemental nickel. The experiment was 84 d in duration and rumen fluid and blood samples were collected on days 42 and 80. Average daily gain and feed efficiency were not affected by nickel supplementation. The addition of 5 ppm supplemental nickel greatly increased rumen bacterial urease activity regardless of protein source. When samples were collected prior to feeding on day 80, nickel increased serum urea nitrogen concentrations in steers fed urea, but decreased circulating urea concentrations in animals fed blood meal or the blood meal-urea combination.Ad libitum intake of trace mineral salt was greatly reduced in steers receiving 5 ppm supplemental nickel. The present study suggests that the source of protein may influence ruminant responses to dietary nickel.     

Renal Excretion of Urea in Reindeer Effect of Nutrition

Renal reabsorption of urea was studied in 8 9-month old reindeer calves fed low protein (19–34 g crude protein, mostly lichens) and high protein (68–69 g crude protein, lichens + soybean meal) diets. Low protein diets fed for a 3-month period resulted in an average renal reabsorption of 93% of the filtered urea, while only 50 % was reabsorbed on the high protein ration. It was calculated that if the reabsorbed urea was used completely for protein synthesis in the rumen, 4 g crude protein could be made daily in the lichen fed animals. This amount would be a very significant contribution to the nitrogen economy of animals which are usually in a negative nitrogen balance when lichens are the main food consumed.     

Dietary arginine degradation is a major pathway in ureagenesis in juvenile turbot (Psetta maxima)

Recent studies indicate that urea excretion is responsive to protein intake and that turbot, Psetta maxima, appear to differ from other species by their urea excretion pattern and levels. This study was undertaken to evaluate the influence of dietary nitrogen and arginine on ureagenesis and excretion in turbot. Juvenile turbot (29 g) were fed semi-purified diets containing graded levels of nitrogen (0-8% dry matter) and arginine (0-3% dry matter) for 6 weeks. Growth data showed that turbot have high dietary nitrogen (123 mg/kg metabolic body weight/day) and very low dietary arginine (9.3 mg/kg metabolic body weight/day) requirements for maintenance. Requirements for unit body protein accretion were 0.31 g and 0.15 g for nitrogen and arginine respectively. Post-prandial plasma urea levels and urea excretion rates showed that urea production was significantly (P<0.05) influenced by dietary arginine levels. While hepatic arginase (EC 3.5.3.1) activity increased significantly (P<0.05) with increasing dietary arginine levels, activities of other enzymes of the ornithine urea cycle were very low. Our data strongly suggest that the ornithine urea cycle is not active in the turbot liver and that dietary arginine degradation is a major pathway of ureagenesis in turbot.     

Influence of Forced-Feeding of Individual Essential Amino Acids on the Activities of All Urea Cycle Enzymes in Rat Liver

The response of all urea cycle enzymes, i.e. carbamyl phosphate synthetase, ornithine transcarbamylase, argininosuccinate synthetase, argininosuccinase and arginase, has been determined in the liver of protein-depleted young rats which were forcibly fed individual essential l-amino acids along with or without caloric sources. The feeding of individual amino acids produced different effects on the level of each of the enzymes, and generally the response of carbamyl phosphate synthetase, argininosuccinate synthetase, argininosuccinase and arginase was greater than that of ornithine transcarbamylase. Of all the essential amino acids tested tryptophan was most effective on the elevation of these enzymes. Several amino acids, phenylalanine, leucine, threonine and methionine had also somewhat effect on the increase of some enzyme activities, but other amino acids had little or no effect on the response of these enzymes. On the contrary, histidine and lysine caused appreciable decrease of arginase activity. These enzyme activities in rats fed tryptophan alone were extremely higher than those of animals fed it along with caloric sources. The response level of the enzymes was essentially dependent on the tryptophan content in diets under the proper conditions. Tryptophan feeding did not produce any increase in both levels of urine and plasma urea despite the elevation of all urea cycle enzyme activities occured.     

Alterations of energy metabolism induced by hexadecane in mice

Two groups of young male OF-1 mice were fed for 60 days with cafeteria or, as controls, with standard pellet diet respectively. At that time, both groups were daily treated with hexadecane (HDK) on the skin. HDK induced a drastic body weight loss much higher in cafeteria than control mice. White adipose tissue were exhausted after 4 days of treatment in controls but not after 10 days in cafeteria ones. HDK resulted in mobilization of liver glycogen in both groups while muscle glycogen decreased slightly in the end. Hexadecane treatment did not result in massively enhanced nitrogen metabolism, as the actual oxidation of amino acids decreased considerably as indicated by the low levels of plasma urea. The results could be explained by powerful and lasting effects of hexadecane on thermogenesis and metabolic reserve balance. The use of this material for pharmacological manipulation of body weight appeared difficult.     

The effect of dietary protein on the amino acid supply and threonine metabolism in the pregnant rat

To characterise the effects of dietary protein content on threonine metabolism during pregnancy, rats were fed diets containing 18% or 9% protein and then killed at different stages of gestation. Serum threonine concentrations fell significantly faster in the animals fed the diet containing 9% protein when compared to those fed the diet containing 18% protein. On day 4 of gestation the rate of threonine oxidation was higher in maternal liver homogenates prepared from the animals fed the diet containing 18% protein. The rate of threonine oxidation by liver homogenates fell as gestation proceeded in both diet groups. The activity of threonine dehydrogenase in the maternal liver was unaffected by dietary protein content at all stages of gestation. Serine-threonine dehydratase activity in homogenates of the maternal liver was transiently increased during the early stages of gestation in the animals fed high protein diets but was unchanged in the low protein groups. There was an increase in serine-threonine dehydratase activity in the kidney during the later stages of gestation but this was unaffected by the protein content of the maternal diet. These data show that the changes in free threonine concentrations cannot be accounted for through changes in the oxidation rate and suggest that some other factor influences the unusual metabolism of this amino acid during gestation.     

Evaluation of corn gluten meal with urea as a source of supplementary nitrogen for growing calves and lambs

Three growth trials and a metabolism study were conducted to evaluate corn gluten meal—urea (CGM—urea) combinations compared to a urea control as sources of supplementary nitrogen for growing calves and lambs fed on diets based on corn cobs.In the first growth study cattle were fed individually twice daily while in the sheep growth study and the second cattle growth study, animals were fed in groups. The average daily gain and feed conversion were similar for animals given the soya bean meal (SBM) and CGM—urea supplements. Feed intakes were similar for all treatments except for growth study 1. Nitrogen retention and dry matter and protein digestibility did not differ significantly between the SBM and CGM—urea supplemented groups. Urea consistently gave inferior results in all the experiments. These results suggest that CGM—urea combination is essentially equal to SBM in supporting growth of calves and lambs.     

Perturbations in nitrogen metabolism of brain and liver of rat following repeated benthiocarb administration

Effects of repeated administration of benthiocarb on the nitrogen metabolism of hepatic and neuronal systems have been studied. Repeated benthiocarb treatment was associated with significant decrease in proteins with a concomitant increase in free amino acids (FAA) and specific activity levels of proteases suggesting impaired protein synthesis or elevated proteolysis. The glycogenic aminotransferases showed a significant elevation in both the tissues indicating high feeding of ketoacids into oxidative pathway for efficient operation of TCA cycle to combat energy crisis during induced benthiocarb stress. However, the activity levels of branched-chain aminotransferases decreased suggesting their reduced contribution of intermediates to TCA cycle. A comparative evaluation of the activity levels of ammonogenic enzymes, AMP deaminase, adenosine deaminase and glutamate dehydrogenase (GDH) indicated that ammonia was mostly contributed by nucleotide deamination rather than by oxidative deamination. GDH exhibited reduced activity due to low availability of glutamate. In accordance with increased levels of urea, the activity levels of arginase, a terminal enzyme of urea cycle was increased suggesting increased urea cycle operation in order to combat the increased ammonia content. As the presence of urea cycle in the brain is rather doubtful, the conversion of ammonia to glutamine for the synthesis of GABA is envisaged in brain whereas in liver, excess ammonia was converted to urea through ornithine-arginine reacting system. The increased glutaminase activity observed during benthiocarb intoxication is accounted for counteracting acidosis or maintenance of metabolic homeostasis. Arginase, a terminal enzyme of ornithine cycle showed increased activity denoting the efficient potentiality of tissues to avert ammonia toxicity. The changes observed in tissues of rat administered with benthiocarb reflects a shift in nitrogen metabolism for efficient mobilization of end products of protein catabolism.     

Effect of developmental stage on arginase and urea production in the liver of rat

Arginase activity in the liver, brain, and testis of rats was examined during the different phases of life span. When expressed as specific activity (micromoles of L-arginine hydrolyzed per minute per gram of whole homogenate protein), the enzyme activity in the brain and testis decreased markedly during the early stage of life and stayed low during the remainder of the life span. On the other hand, the arginase in the liver showed a great dependency on the developmental phase of the animal, showing two distinct peaks: one during the early phase (20 days after birth) and the other at a later time (3 months of age). This pattern of change in the hepatic arginase activity closely coincided with the pattern of the rate of urea synthesis determined with liver slices and expressed in terms of micromoles of urea formed per hour per gram of tissue slice. In contrast to the above observations, however, curves obtained by plotting the total liver arginase or urea synthetic activity vs the developmental stage of rats showed no measurable discontinuity. Further studies revealed that the observed pattern of specific activity of hepatic arginase was, in part, due to the change in the relative concentration of arginase protein in the liver.     

The dogfish shark (Squalus acanthias) increases both hepatic and extrahepatic ornithine urea cycle enzyme activities for nitrogen conservation after feeding

Urea not only is utilized as a major osmolyte in marine elasmobranchs but also constitutes their main nitrogenous waste. This study investigated the effect of feeding, and thus elevated nitrogen intake, on nitrogen metabolism in the Pacific spiny dogfish Squalus acanthias. We determined the activities of ornithine urea cycle (O-UC) and related enzymes in liver and nonhepatic tissues. Carbamoyl phosphate synthetase III (the rate-limiting enzyme of the O-UC) activity in muscle is high compared with liver, and the activities in both tissues increased after feeding. The contribution of muscle to urea synthesis in the dogfish body appears to be much larger than that of liver when body mass is considered. Furthermore, enhanced activities of the O-UC and related enzymes (glutamine synthetase, ornithine transcarbamoylase, arginase) were seen after feeding in both liver and muscle and were accompanied by delayed increases in plasma urea, trimethylamine oxide, total free amino acids, alanine, and chloride concentrations, as well as in total osmolality. The O-UC and related enzymes also occurred in the intestine but showed little change after feeding. Feeding did not change the rate of urea excretion, indicating strong N retention after feeding. Ammonia excretion, which constituted only a small percentage of total N excretion, was raised in fed fish, while plasma ammonia did not change, suggesting that excess ammonia in plasma is quickly ushered into synthesis of urea or protein. In conclusion, we suggest that N conservation is a high priority in this elasmobranch and that feeding promotes ureogenesis and growth. Furthermore, exogenous nitrogen from food is converted into urea not only by the liver but also by the muscle and to a small extent by the intestine.     

谷氨酰胺对创伤后大鼠恢复的效果

本研究观察了谷氨酰胺（ＧＬＮ）对创伤大鼠恢复的影响,１７只ｗｉｓｔａｒ大鼠随机分成ＧＬＮ组（９只）和对照组（８只）,大鼠行创伤手术后,分别饲喂含或不含１．２％ＧＬＮ的合成饲料。实验中观察了动物体重恢复、血浆蛋白变化,实验第１４天,处死动物,测定伤口皮肤抗张力强度和胶原蛋白含量、淋巴细胞转化率、小肠粘膜核酸及蛋白质含量。用Ｓｔｕｄｅｎｔ＇ｓＴ检验分析结果差异的显著性。实验结果显示：两组动物的饲料摄入量、体重增长、血浆蛋白水平、氮平衡均无明显差别,但在实验结束时,ＧＬＮ组大鼠的小肠粘膜ＤＮＡ、ＲＮＡ及蛋白质含量显著高于对照组（Ｐ＜０．０５）,同时其伤口皮肤抗张力强度、伤口胶原蛋白合成以及反映机体免疫功能的淋巴细胞转化率,亦有高于对照组的趋势,但无统计学差别（Ｐ＞０．０５）,揭示了ＧＬＮ具有保护肠道粘膜的作用,有益于大鼠创伤后的恢复。     

Urea formation by postoperative liver

The effect of liver left lobe resection (LR, 15–20% of the organ weight) on hepatic urea formation was investigated in 84 albino rats. The objects used for analysis of urea content included: operated left lobe (LLL), non-operated middle (MLL) lobe of the liver, blood (aorta, v. hepatica, v. porta), and choledochal bile. Arginase activity was examined in liver homogenate. On the day 3 and day 7 after resection reduced arginase activity was detected. LR caused a decrease in urea content in v. hepatica, but increased urea content in the arterial blood and v. porta. The increase in bile urea on day 7 was then changed for its decrease observed on day 14 of the postoperative period. The urea content in the liver on day 3 after LR was below the norm, while on days 7 and 14 it became normal. Results of this study suggest impaired urea formation by hepatocytes of the operated liver and activation of extrahepatic mechanisms of urea formation from arginine.     

大豆寡糖对断奶环江香猪生长性能和营养物质代谢的影响

为了探讨大豆寡糖(SBOS)作为抗生素替代物对断奶仔猪生长性能的影响及其作用机制,饲养试验选用21日龄断奶的环江香猪12头,随机分为2组,每组6头,分别饲喂添加0.5%SBOS日粮和抗生素对照日粮,试验期为14 d。记录每天的采食量,分别于试验开始和结束时称取空腹体重并采集血液,肝素抗凝离心分离血浆,测定生化参数和游离氨基酸水平。消化试验选用10头环江香猪,随机分为2组,处理同饲养试验。预试期4 d,正试期3 d,收集粪样,指示剂法测定干物质、粗蛋白和粗脂肪的消化率。结果表明,与抗生素对照组相比,SBOS组仔猪平均日采食量、日增重和料重比均无显著变化;血浆尿素氮和血氨含量显著下降,总蛋白含量显著升高(P<0.05);血浆甘氨酸、组氨酸、赖氨酸、精氨酸和蛋氨酸的含量均显著升高(P<0.05);粗蛋白和粗脂肪消化率有提高趋势。提示SBOS可替代抗生素用于断奶仔猪日粮。     

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.     

Effects of dietary salt intake on plasma arginine

Because L-arginine is degraded by hepatic arginase to ornithine and urea and is transported by the regulated 2A cationic amino acid y(+) transporter (CAT2A), hepatic transport may regulate plasma arginine concentration. Groups of rats (n = 6) were fed a diet of either low salt (LS) or high salt (HS) for 7 days to test the hypothesis that dietary salt intake regulates plasma arginine concentration and renal nitric oxide (NO) generation by measuring plasma arginine and ornithine concentrations, renal NO excretion, and expression of hepatic CAT2A, and arginase. LS rats had lower excretion of NO metabolites and cGMP, lower plasma arginine concentration (LS: 83 +/- 7 vs. HS: 165 +/- 10 micromol/l, P < 0.001), but higher plasma ornithine concentration (LS: 82 +/- 6 vs. HS: 66 +/- 4 micromol/l, P < 0.05) and urea excretion. However, neither the in vitro hepatic arginase activity nor the mRNA for hepatic arginase I was different between groups. In contrast, LS rats had twice the abundance of mRNA for hepatic CAT2A (LS: 3.4 +/- 0.4 vs. HS: 1.6 +/- 0.5, P < 0.05). The reduced plasma arginine concentration with increased plasma ornithine concentration and urea excretion during LS indicates increased arginine metabolism by arginase. This cannot be ascribed to changes in hepatic arginase expression but may be a consequence of increased hepatic arginine uptake via CAT2A.