Threonine imbalance, deficiency and neurologic dysfunction in the kitten

The effect of feeding threonine-imbalanced and threonine-deficient purified diets (containing L-amino acids as the only source of dietary nitrogen on food intake, weight gain and blood plasma amino acid pattern has been examined in growing kittens. The imbalance was created by adding 17.5% of amino acid mixture lacking threonine to a low amino acid basal diet comtaining 17.5% amino acid mixture including 0.4% threonine. A depression in food intake and weight gain occurred while feeding the imbalanced diet which was corrected by adding an additional 0.2% threonine to the imbalanced diet. There was no adaptation with time in the form of increased food intake or weight gain while feeding the imbalanced diet. Plasma threonine was consistently and similarly depressed (10 to 35% of normal) while feeding the basal, imbalanced and corrected diets and increased to normal when the standard diet with 1.4% threonine was fed. Plasma threonine and total amino acid concentrations of kittens fed the imbalanced diet did not differ from those observed in kittens fed the basal diet. Signs of neurologic dysfunction and/or lameness developed in 14 of the 17 kittens fed threonine-imbalanced or deficient diets, which resolved as dietary threonine was increased.     

Temporal response of hepatic threonine dehydrogenase in chickens to the initial consumption of a threonine-imbalanced diet

Amino acid imbalances contribute to higher requirements of amino acids than would occur if the dietary profile of amino acids perfectly matched the requirements. The mechanisms of imbalances have not been fully elucidated. Because threonine dehydrogenase (TDH) activity in liver mitochondria increases in chicks and rats subjected to threonine imbalance, the current study was carried out to determine whether the change in TDH activity occurs rapidly enough after the consumption of an imbalanced diet to be considered a possible primary metabolic response. In a series of experiments, Leghorn chicks were allowed free access to a semipurified basal diet marginally limited in threonine or the same diet containing a mixture of indispensable amino acids (IAA) lacking threonine to cause a threonine imbalance. In the first experiment, dietary supplements of 5.5 and 11.1% IAA were used to determine a level of supplement that would cause a robust response in the specific activity of TDH. Feed intake, body weight gains and efficiency of feed utilization were lower and specific activities of TDH were higher in chicks fed 11.1% IAA than in those fed 5.5% IAA. In subsequent experiments, hepatic TDH activities and plasma amino acid profiles of the control and experimental groups were determined at 1. 5, 3, 6, 12 and 24 h after the first offering of the diet containing 11.1% IAA. The specific activities of TDH in chicks fed the IAA supplement were 40-150% higher (P < 0.05) and plasma threonine concentrations were 42-53% lower (P < 0.05) than in chicks fed the basal diet at all times except 1.5 h. These results indicate that changes in the capacity for threonine degradation via TDH may occur in the liver within a few hours after the consumption of a threonine-imbalanced diet and suggest the possibility that altered TDH activity may contribute to the increased threonine requirement associated with threonine imbalance.     

Effect of an amino acid imbalance on intestinal sucrase and leucine aminopeptidase activities in rats.

In order to investigate the relationship between dietary amino acids and protein, as well as the activities of intestinal sucrase and leucine aminopeptidase in rats, the effects of an amino acid imbalance on these enzyme activities were studied. The amino acid imbalance was created by adding 8% of an indispensable amino acid mixture lacking threonine to a 6% casein diet supplemented with 0.3% methionine. The food intake and growth of rats fed the imbalanced diet ad libitum were depressed, and the segmental weights of the small intestine and its sucrase activity were clearly lower than those of rats fed the basal diet. The effect of the imbalanced diet under pair-feeding condition on the sucrase activity was similar to that under an ad libitum feeding condition. The food intake and segmental sucrase activity, that is, sucrase activity per length of the small intestine, of rats injected with cortisol (1 mg/day) and fed the imbalanced diet were not depressed, although administration of insulin (1.5 U/day) had no effect on the food intake or segmental sucrase activity. Force-feeding stimulated growth of rats receiving the imbalanced diet, as well as increasing their segmental sucrase activities. The effects of these different conditions on the leucine aminopeptidase activity of rats receiving the imbalanced diet were obscure. These results suggest that changes in segmental sucrase activity might be mediated by stimulating factors in food intake affected by the composition of ingested amino acids and protein together with sucrose in the gastrointestinal lumen.     

Effects of amino acid imbalance and protein content of diets on food intake and preference of young, adult, and diabetic rats.

Effects of histidine or methionine imbalance and dietary levels (3-50%) of casein on food intake and preference of young, adult, and diabetic (2.5 month old) rats were examined. Depressions in food intake and growth caused by ingestion of the imbalanced diet were greatest in young rats and least or absent in diabetic rats. Alloxan diabetes induced hyperphagia and elevated concentrations of plasma branched-chain amino acids and decreased concentrations of tryptophan and tyrosine. The diabetic rats fed the imbalanced diet for 9 days had a higher concentration of the limiting amino acid in the plasma than the adult normal rats fed the same diet. The diabetic rats preferred the imbalanced diet over a protein-free diet when they were fed these diets concurrently. Ingestion of the imbalanced diet by normal rats caused greater changes in plasma and brain amino acid patterns than did the protein-free diet. Unlike the diabetic rats, the normal rats, especially the young rats, strongly preferred the protein-free diet over the imbalanced diet. The normal rats also preferred a 10% casein diet supplemented with L-methionine over a low or high casein diet. It seemed that young rats were able to select a protein diet that supported maximal growth when proportions of dietary amino acids were balanced. It also seemed that the susceptibility of the rats to amino acid imbalance varied directly with the status of overall protein synthesis of the animals.     

Involvement of food intake in the lysine-arginine antagonism in chicks.

Studies were conducted to evaluate the involvement of food intake in the lysine-arginine antagonism. Diets were formulated to compensate for the metabolic consequences of excess dietary lysine; induction of renal arginase activity, depression of heptic glycine transamidinase, and urinary losses of arginine. This was accomplished by inclusion of creatine in the basal diet, use of a moderate excess of lysine that did not increase urinary arginine excretion, and addition of the arginase depressors, alpha-aminoisobutyric acid (AIB) and L-threonine, to diets containing excess lysine. When chicks were fed diets containing excess lysine ad libitum, growth and efficiency of arginine retention were reduced. Supplementation of the diets with AIB and threonine markedly reduced the growth depression and restored efficiency of arginine utilization. When chicks were force-fed the diet containing excess lysine, growth was depressed, and body composition was altered. Inclusion of AIB and threonine in the diet containing excess lysine resulted in growth and body composition equivalent to levels of force-fed controls. In a second experiment the basal diet and basal supplemented with AIB and threonine were pair-fed to lysine-supplemented diets containing AIB and threonine. Body weight gains and body composition of all groups were similar. In other experiments, food intake increased within 24 hours (P less than 0.05) and probably within 12 hours (P less than 0.10) after removal of excess lysine from the diet. It is concluded that a portion of the lysine-arginine antagonism is due to a primary effect of lysine on regulation of food intake.     

The branched-chain amino acid antagonism in chicks

The effects of dietary supplements of branched-chain amino acids on growth, food consumption and metabolism in chicks were investigated. When an adequate diet contained 1.20, 1.60, 2.25, 3.75, or 5.00% leucine, increasing leucine content caused reduced food consumption and weight gains, coupled with impaired efficiency of food utilization. When the diet deficient in branched-chain amino acids contained 0.98, 1.46, 2.25, 3.75, or 5.00% leucine, increasing leucine resulted in increased food consumption and reduced efficiency of food utilization when levels of leucine up to 3.75% were fed. Excess leucine depressed plasma concentrations of isoleucine and valine. Excesses of isoleucine or valine caused smaller depressions of concentrations of the other two branched-chain amino acids. All these effects were seen during the first 8 days of experiment, after which they diminished or disappeared. Muscle branched-chain amino acid aminotransferase (BCAT) (L-leucine:2-oxoglutarate aminotransferase, EC 2.6.1.6) activity was increased in chicks fed excess leucine but not in those fed excess isoleucine or valine. Hepatic alpha-ketoisocaproic dehydrogenase (KADH) (2-oxoisocaproate:lipoate oxidoreductase, EC 1.2.4.3) activity and muscle polyribosomal aggregation were unaffected by diet. When chicks were fed diets containing either 0.98 or 2.25% leucine, production of 14CO2 from [1-14C]isoleucine and [1-14C]valine was increased in chicks fed the higher level of leucine. The increase was small in both cases, representing approximately 2% of consumed isoleucine and valine. Increased production of 14CO2 was observed within 12 hours of feeding excess leucine; however, BCAT increased only after 2 to 4 days. No differences were seen in excreted 14C or in the relative distribution of 14C along the small intestine. We conclude that the chick is able to adapt in part to excesses of dietary leucine and that the branched-chain amino acid antagonism may involve increased catabolism of the limiting branched-chain amino acids.     

Influence of dietary potassium on lysine metabolism in the chick

High levels of dietary cations have been reported to spare the arginine requirement of chicks fed a diet containing excess lysine. The present studies were conducted to determine the nature of this effect, using potassium as the variable cation. The inclusion of 1.8% potassium in a high lysine diet improved growth but did not affect consistently the efficiency of feed utilization, nor did it have a significant effect on arginine metabolism as measured by renal arginase activity, urea excretion or arginine excretion. Lysine catabolism was markedly increased, however. Chicks fed the potassium supplemented diet converted approximately 23% of an oral loading dose of 14C lysine to respiratory 14CO2 over a 6-hour period, in contrast to approximately 9% of the dose in chicks fed the unsupplemented diet containing 0.4% potassium. The activity of hepatic lysine-alpha-ketoglutarate reductase, an initial enzyme in the catabolism of lysine, was increased in response to potassium or arginine supplementation. It is proposed that high levels of potassium or other cations may improve the growth of chicks fed diets containing excess lysine by increasing lysine catabolism. The effect on growth may be mediated primarily by increasing food intake and to a lesser extent by alleviation of the other metabolic manifestation of the lysine-arginine antagonism.     

Histidine-imbalanced diets stimulate hepatic histidase gene expression in rats.

A high protein concentration in the diet induces the gene expression of several amino acid degrading enzymes such as histidase (Hal) in rats. It is important to understand whether the amino acid pattern of the dietary protein affects the gene expression of these enzymes. The purpose of the present work was to study the effect of a histidine-imbalanced diet on the activity and mRNA concentration of rat hepatic histidase. Seven groups of six rats were fed one of the following diets: 1) 6% casein (basal), 2) 20% casein, 3) 35% casein, 4) an imbalance diet containing 6% casein plus a mixture of indispensable amino acids (IAA) equivalent to a 20% casein diet without histidine (I-20), 5) 6% casein plus a mixture of IAA equivalent to a 35% casein diet without histidine (I-35), 6) a corrected diet containing 6% casein plus IAA including histidine equivalent to a 20% casein diet, 7) a corrected diet containing 6% casein plus IAA including histidine equivalent to a 35% casein diet. Serum histidine concentration was inversely proportional to the protein content of the diet, and it was significantly higher in rats fed the corrected diets compared to their respective imbalanced diet groups. Hal activity increased as the protein content of the diet increased. Greater histidine imbalance resulted in lower food intake and higher Hal activity. Rats fed histidine-corrected diets had lower activity than their respective imbalanced groups. Differences in Hal activity were associated with differences in the concentration of Hal mRNA. These results indicate that rats fed a histidine-imbalanced diet exhibit reduced food intake and weight gain and increased Hal gene expression as a consequence of an increased amino acid catabolism.     

Threonine metabolism in vivo: effect of threonine intake and prior induction of threonine dehydratase in rats

The metabolic fate of threonine was investigated in young male rats fed 15% amino acid diets containing from 0.15% to 0.85% of L-threonine. Liver serine-threonine dehydratase (S-TDH) activity did not increase with increasing dietary threonine content. The level of threonine required for maximum weight gain was not greater than 0.55% of the diet (or about 600 mumoles/day). Tissue free threonine content of rats fed the diets with 0.15% or 0.3% of threonine was very low but increased sharply with increasing dietary threonine content above 0.3%. During ad libitum feeding of these diets containing L-[U-14C]threonine, rate of oxidation of threonine was low when intake was in the range of the requirement for maximum growth, but increased, thereafter as threonine intake increased. A 30-fold induction of liver S-TDH, by prior feeding of an 80% casein diet, did not result in increased oxidation of threonine when dietary threonine content was 0.15%. When dietary threonine content was increased to 0.5%, oxidation of threonine increased slightly but significantly. With 3% of threonine in the diet, rats previously fed a 15% casein diet had extremely high tissue threonine concentrations whereas those with high S-TDH activity, due to the previous feeding of the 80% casein diet, oxidized threonine rapidly and tissue threonine concentrations were elevated much less.     

Threonine imbalance and the threonine requirement of the chicken

Experiments were conducted to determine the influence of dietary amino acids on the threonine requirement of chicks. A diet limiting in threonine and containing the equivalent of 20.6% crude protein was imbalanced by supplements of 0.9 to 1.5% L-tryptophan, 3.0% L-serine, a mixture of 2% each, leucine, isoleucine and valine or a 6.0% mixture of all essential amino acids but threonine. All amino acid supplements decreased the growth of chicks, and decreased food intake and/or efficiency of food utilization (P less than 0.05). Supplemental threonine prevented these effects in all cases except for 1.5% tryptophan in which the adverse effect of tryptophan was not prevented completely. The threonine requirement of Leghorn chicks was 0.69 to 0.72 percent of the diet when the diet did not contain the imbalancing amino acid supplements. We concluded that the threonine requirement of the chick is higher than previously reported, and is influenced by the amino acid content of the diet.     

Threonine concentration in the prepyriform cortex has separate effects on dietary selection and intake of a threonine-imbalanced diet by rats

Dietary selection and intake of a threonine-imbalanced diet were evaluated after increasing the concentration of the dietary limiting amino acid in the prepyriform cortex. Selection against the threonine-imbalanced diet in favor of a protein-free diet was reversed when 2 or 4 nmol threonine was injected bilaterally into the prepyriform cortex. However, intake of the threonine-imbalanced diet was significantly increased only after injection of 2 nmol threonine. The reduced intake of the threonine-imbalanced diet, compared to the basal diet, after injection of 4 nmol threonine was not the result of an excess in the concentration of amino acid (or nitrogen) injected into the PPC because intake of the threonine-basal diet was not reduced when 4 nmol threonine was administered. Intake of the threonine-imbalanced diet was also increased after injection of 2 nmol threonine plus 2 nmol isoleucine but not after injections of 2 nmol isoleucine. The changes in food intake when an imbalanced diet is fed appear to be the result of at least two separable responses: recognition of a diet as having an amino acid imbalance, as indicated by dietary choice, and reduction in food intake. The results of this study indicate that changing the concentration of the dietary limiting amino acid in the prepyriform cortex influenced dietary selection and food intake separately.     

Dietary disproportions of amino acids in the rat: effects on food intake, plasma and brain amino acids and brain serotonin.

Food intake, growth, plasma and brain amino acid, and brain serotonin and 5-hydroxyindole-3-acetic acid (5-HIAA) concentrations were measured in rats fed low protein diets containing disproportionate amounts of large neutral amino acids (LNAA) devoid of tryptophan or histidine (tryptophan or histidine imbalance). Five-day food intakes and weight gains of rats fed the imbalanced diets were depressed. The concentration of the limiting amino acid was low in brains of rats fed diets containing LNAA that compete with either tryptophan or histidine for entry into brain. Correlations were observed between the brain concentrations of most individual LNAA and either the ratios of the plasma concentration of that LNAA to the sum of the other LNAA, or the predicted rates of influx of that LNAA. Cumulative food intakes were correlated with brain concentrations of the limiting amino acid, tryptophan or histidine. Food intakes were not consistently correlated with concentrations of serotonin and 5-HIAA because these compounds were altered only in brains of rats in the tryptophan study. Competition among amino acids for uptake into brain appears to be involved in the feeding response of the rat to dietary disproportions of amino acids, but this response is not directly related to changes in brain concentrations of serotonin and 5-HIAA.     

Adaptation of rats to diets containing different levels of protein: effects on food intake, plasma and brain amino acid concentrations and brain neurotransmitter metabolism

Food intake, plasma and brain amino acid concentrations, liver amino acid catabolic enzyme activities, and whole-brain neurotransmitter and metabolite concentrations were measured in young rats adapted for 11 d to diets containing from 5 to 75% (in increments of 5%) casein. Food intake was depressed initially in rats fed diets containing 5, 10% or greater than 35% casein. For the duration of the experiment, food intakes of the groups fed the higher protein diets improved on successive days; the length and severity of the depression were proportional to the protein content of the diet fed. Rats fed low levels of protein grew poorly, and their food intake remained depressed. The gradual improvement in growth and food intake of rats fed diets containing more than 35% casein was accompanied by dramatic increases in the activities of serine-threonine dehydratase (SDH, EC 4.2.1.16) and glutamate-pyruvate aminotransferase (GPT, EC 2.6.1.1) in liver. The increase in amino acid catabolic activity was accompanied by decreases in the concentrations of most amino acids in plasma and brain. However, concentrations of branched-chain amino acids, in both plasma and brain, increased in direct proportion to the protein concentration of the diet fed. As a result of these reciprocal responses, the total concentration of indispensable amino acids in brain (IAA) was maintained within a narrow range of values, despite a sixfold range of protein intakes. Whole-brain concentrations of norepinephrine, dopamine and serotonin were not correlated with dietary protein concentration, total food intake or protein intake. Brain concentrations of homovanillic acid and 5-hydroxyindoleacetic acid were correlated inversely with protein intake and that of 3,4-dihydroxyphenylacetic acid was correlated directly with food intake. Protein intake appeared to be related to the animal's ability to maintain brain total IAA content between some upper and lower limits. Our results indicate that this was accomplished initially through downward adjustment of protein intake and subsequently through an increase in catabolic capacity for the amino acids.     

Induction of lysine imbalance in rats: relation to competition for lysine transport into the brain in vitro

The ability of amino acids inhibitory of lysine transport into brain slices to induce lysine imbalance was determined by feeding wheat gluten or casein diets with additions of such amino acids. Lysine transport was only moderately inhibited by amino acids; the most effective were basic amino acids or mixtures of indispensable (IAA) or branched chain amino acid (BCAA). Only mild depressions in growth and food intake occurred during a 10-day period when male, 60--65 g rats of the Sprague-Dawley strain were fed lysine-limiting, 18% wheat gluten diets with additions of these amino acids. The effects were prevented by added lysine. Rats allowed a choice between the lysine-imbalanced or non-protein diets selected the imbalanced, wheat gluten diets (in severe imbalances rats will choose the non-protein diet). Growth depression, prevented by added lysine, occurred in rats fed a 6% casein diet supplemented with IAA; individual amino acids were ineffective. Growth depressions also occurred when rats were fed a basal diet containing 6% case in + 5% of an equimolar mixture of nine IAA and supplemented with arginine or more IAA; BCAA were less effective. Additional lysine completely prevented the growth depressions, but growth of rats fed the diets containing arginine and BCAA was greater than that of those fed the extra IAA. It is difficult to induce a severe lysine imbalance; this is consistent with the failure of amino acids to cause under our conditions strong inhibition of lysine transport into brain.     

Induction of threonine imbalance by dispensable amino acids: relation to competition for amino acid transport into brain.

The ability of low protein diets containing small neutral, dispensable amino acids to induce threonine imbalance has been examined. Diets containing amino acids which compete for threonine transport in vitro (serine, alanine, alpha-amino-n-butyrate) caused depressions of growth and food intake which could be corrected to varying degrees by adding threonine to the diet. Large neutral, indispensable amino acids, moderately inhibitory of threonine transport, also induced the imbalance. Some amino acids that had little or no effect on threonine transport in vitro (acidic amino acids and proline) did not cause growth and food intake depressions. Other non-inhibitory amino acids (arginine and lysine) caused growth depressions which were not satisfactorily corrected by additional threonine alone, but were prevented by supplements of all the indispensable amino acids including threonine. Ornithine which was also not inhibitory of threonine transport was an exception. It induced a moderate growth depression which was corrected by additional threonine. Similar studies showed that histidine or tryptophan imbalance could be induced by feeding diets containing only those large neutral amino acids which compete for histidine or tryptophan transport in vitro. These experiments show that, based on the results of transport competition experiments, it is generally possible to devise amino acid supplements which can induce a dietary imbalance of a given amino acid.     

The effects of maternal protein restriction on the growth of the rat fetus and its amino acid supply.

Maternal protein deficiency causes fetal growth retardation which has been associated with the programming of adult disease. The growth of the rat fetus was examined when the mothers were fed on diets containing 180, 90 and 60 g protein/kg. The numbers of fetuses were similar in animals fed on the 180 and 90 g protein/kg diets but the number was significantly reduced in the animals fed on the 60 g protein/kg diet. The fetuses carried by the mothers fed on the 90 g protein/kg diet were 7.5% heavier than those of mothers fed on 180 g protein/kg diet on day 19 of gestation, but by day 21 the situation was reversed and the fetuses in the protein-deficient mothers were 14% smaller. Analysis of the free amino acids in the maternal serum showed that on day 19 the diets containing 90 and 60 g protein/kg led to threonine concentrations that were reduced to 46 and 20% of those found in animals fed on the control (180 g/kg) diet. The other essential amino acids were unchanged, except for a small decrease in the branched-chain amino acids in animals fed on the 60 g protein/kg diet. Both low-protein diets significantly increased the concentrations of glutamic acid+glutamine and glycine in the maternal serum. On day 21 the maternal serum threonine levels were still reduced by about one third in the group fed on the 90 g protein/kg diet. Dietary protein content had no effect on serum threonine concentrations in nonpregnant animals. Analysis of the total free amino acids in the fetuses on day 19 showed that feeding the mother on a low-protein diet did not change amino acid concentrations apart from a decrease in threonine concentrations to 45 and 26% of the control values at 90 and 60 g protein/ kg respectively. The results suggest that threonine is of particular importance to the protein-deficient mother and her fetuses. Possible mechanisms for the decrease in free threonine in both mother and fetuses and the consequences of the change in amino acid metabolism are discussed.     

Effect of dietary asparagine and protein-equivalents in crystalline amino acid diets on asparagine metabolism in chicks

The effect of dietary asparagine and protein-equivalents from crystalline amino acid mixtures upon asparagine metabolism in chicks were studied. Liver and kidney asparaginase activities were significantly increased in chicks fed 44.6% protein-equivalents compared to chicks fed the Illinois chick standard amino acid mixture containing 14.8% protein-equivalents. The asparagine synthetase activity in chick liver and kidney was not significantly changed by protein-equivalents or dietary asparagine. Liver and kidney asparaginase activities in chicks fed 14.8% protein-equivalent standard diets were decreased with increasing levels of dietary asparagine (0,2 and 6%). Kidney asparaginase activities in chicks fed 44.6% protein-equivalents also were decreased with increasing levels of asparagine but liver asparaginase in these chicks was not changed with dietary asparagine. The plasma asparagine concentration was dependent on the amount of dietary asparagine and protein-equivalents. Dietary asparagine increased plasma asparagine in chicks fed 14.8 and 44.6% protein-equivalent diets but plasma asparagine in chicks fed the 14.8% protein-equivalent diet plus 6% asparagine was 3.5 times higher than plasma asparagine in chicks fed the diet containing 44.6% protein-equivalent plus 6% dietary asparagine. Plasma asparagine in chicks fed the 44.6% protein-equivalent diet with 6% asparagine was reduced due to increased asparaginase activity.     

Studies on the severity of various amino acid imbalances in the young male rat.

Relative severities of amino acid imbalance were compared in young male rats fed diets having an amino acid imbalance with respect to various essential amino acids. In the imbalanced diets, one or more essential amino acids were made to be low, ranging from suboptimal to two times the requirement, while the other essential amino acids were present at three times the requirement. From the extent of depressions in food intake and growth, the severity of amino acid imbalance was found to be dependent upon the level as well as the type of the limiting amino acid(s) in each imbalanced diet. The overall results indicated that Met imbalance was very severe, followed in decreasing order by Ile, Trp, Leu and Val, His, Phe, Thr, Lys and Arg imbalance. The imbalance with respect to the large neutral amino acid appeared to be more severe than the imbalance with respect to the small neutral amino acid, while the imbalance with respect to the basic amino acid was least severe.     

Barley beta-glucans alter intestinal viscosity and reduce plasma cholesterol concentrations in chicks.

Ninety-six 14-d-old male broiler chicks were divided into three dietary groups and fed a corn-soybean meal diet, a barley diet with beta-glucanase and that diet without beta-glucanase. All diets contained 4 g cholesterol/kg. Average daily body weight gain, plasma total cholesterol concentration, LDL cholesterol concentration and digestibility of lipids and protein were lowest (P < 0.05) in the chicks fed the barley diet without beta-glucanase and highest (P < 0.05) in the chicks fed corn-soybean meal diet. Supplementation of the barley diet with beta-glucanase resulted in greater (P < 0.05) average daily weight gain, plasma total and LDL cholesterol concentrations and digestibility of lipids. Viscosity of small intestinal digesta was greatest in chicks fed barley, lowest in those fed the corn-soybean diet and intermediate in chicks fed enzyme-treated barley. Significant (P < 0.01) negative correlations occurred between viscosity of the small intestinal contents and average daily weight gain, plasma total and LDL cholesterol concentrations, and digestibility of lipids and protein. A lower concentration of insoluble beta-glucans in small intestinal digesta of the chicks fed barley supplemented with beta-glucanase compared with the chicks fed the unsupplemented barley diet reflects hydrolytic activity of the supplemental beta-glucanase in the diet.     

Dietary protein concentration regulates the mRNA expression of chicken hepatic malic enzyme

Chicken hepatic malic enzyme activity varies with dietary protein content. The mechanisms responsible for this alteration in activity are unclear. In a series of four experiments, broiler chicks were allowed free access for 1.5, 3, 6 or 24 h to a low (13 g/100 g diet), basal (22 g/100 g diet) or high (40 g/100 g diet) protein diet. The diets were isocaloric and had equal concentrations of dietary fat. Hepatic malic enzyme mRNA expression and enzyme activity as well as total liver lipid concentration were examined for each experimental duration. There were no differences in the expression of the mRNA for malic enzyme at 1.5 h, but at 3, 6 and 24 h, malic enzyme mRNA expression was significantly (P < 0.05) reduced in chicks fed the high protein diet and significantly enhanced in chicks fed the low protein diet compared with chicks fed the basal diet. Hepatic malic enzyme activities and total lipid concentration were not different among the chicks fed the different diets at 1.5 and 3 h. At 6 and 24 h, malic enzyme activity and total liver lipid concentration were both significantly greater in birds fed the low protein diet compared with levels in the birds fed the other two diets. In birds fed the high protein diet, malic enzyme activity and total liver lipid concentration were significantly reduced at 24 h compared with birds fed the basal diet. In a final experiment, the observed differences in malic enzyme mRNA expression at 6 h were confirmed when chicks were given access to isocaloric diets with the same protein levels as the initial 4 experiments, but with the dietary concentration of carbohydrate held constant. The results suggest that previously observed alterations in the activity of malic enzyme, which were correlated with dietary protein intake, are due to rapid changes in the mRNA expression of this enzyme.