Phenylalanine-pyruvate aminotransferase activity in chicks subjected to phenylalanine imbalance or phenylalanine toxicity.

Two experiments were done to determine the influence of Phe imbalance and excess on Phe-pyruvate aminotransferase (PAT) activity in the chick. Five replicates of 3 chicks (experiment 1) or 2 chicks (experiment 2) of a commercial brown egg layer strain were fed a semipurified diet for 1 wk and then received experimental diets for 10 d. Three diets were used in experiment 1: the basal diet contained 0.46% Phe; the imbalance diet was similar to the basal diet except that it contained a 10% mixture of indispensable amino acids lacking Phe (IAA - Phe) to create a Phe imbalance; the imbalance corrected diet was similar to the imbalance diet except that it was supplemented with 1.12% Phe to correct the imbalance. A 3 x 2 factorial arrangement of treatments in experiment 2 provided 3 dietary levels (0.46, 1.58, and 2.46%) of Phe and either no supplement or 10% supplement of IAA - Phe. Nonfasted chicks were killed and livers were sampled in experiment 1, and livers, kidneys, brains, and pectoralis major muscles were sampled in experiment 2. In experiment 1, liver PAT activity per gram of liver was 80 and 55% higher (P < 0.01) in chicks fed the imbalance and imbalance corrected diets than in chicks fed the basal diet. In experiment 2, the livers and kidneys, but not brains and muscles, of chicks that received the 10% supplement of IAA - Phe had higher activities of PAT per gram of tissue per minute and per milligram of tissue protein extract per minute than chicks that did not receive IAA - Phe (P < 0.001). No effect of dietary Phe on PAT activity was detected (P > 0.05). Phenylalanine-pyruvate aminotransferase activity appears to be regulated in response to dietary content of indispensable amino acids but not by the dietary level of Phe.

Phenylalanine hydroxylase activity and expression in chicks subjected to phenylalanine imbalance or phenylalanine toxicity.

Experiments were performed to investigate the activity of hepatic Phe hydroxylase (PAH) and plasma amino acid concentrations under conditions of Phe imbalance or toxicity in chicks fed on experimental diets from 7 to 14 or 16 d of age. In experiment 1, Phe imbalance was created by adding 10% of a mixture of indispensable amino acids lacking Phe (IAA - Phe) to a basal diet containing 0.46% Phe. The activity of PAH was not significantly affected by the imbalance. Correcting the imbalance by adding 1.12% Phe to the diet prevented the growth impairment and increased the activity of PAH. In experiment 2, growth was reduced by the addition of excess (2%) Phe to the basal diet. Correcting the excess by adding the IAA - Phe to the diet prevented the growth reduction. The activity of PAH was not significantly affected by 2% Phe, but it increased in chicks fed the corrected diet. The levels of PAH mRNA were not affected by the dietary treatments. A factorial arrangement of treatments with 3 dietary levels of Phe (0.46, 1.58, and 2.46%) with or without the IAA - Phe was used in experiment 3. The effects on growth were similar to those of the same treatments in experiments 1 and 2. The addition of Phe significantly increased hepatic PAH activity, but there was no detectable main effect of the IAA - Phe and no interaction. Plasma Phe concentration was increased by dietary Phe and decreased by the IAA - Phe mixture. We conclude that hepatic PAH activity in chicks variably increases in response to Phe or a 10% dietary supplement of indispensable amino acids including Phe but does not increase in response to IAA - Phe when the amino acids are added to a diet that is marginally adequate in Phe. The increased activity does not involve changes in PAH mRNA. The effects of IAA - Phe on plasma Phe concentrations appear to be independent of hepatic PAH activity as measured in vitro.

Phenylalanine requirement, imbalance, and dietary excess in one-week-old chicks: growth and phenylalanine hydroxylase activity.

Two experiments were performed to study Phe imbalance and toxicity in 1-wk-old Babcock B380 chicks resulting from the addition of either a mixture of indispensable amino acids lacking Phe (IAA - Phe) or excess Phe to a diet that was nutritionally adequate in Phe. Chicks received a preexperimental semipurified diet for 1 wk and experimental diets from 7 to 14 d of age. In the first experiment, the chicks were given diets with Phe levels at 0.24, 0.29, 0.34, 0.39, 0.44, and 0.49% of the diet to determine the Phe requirement. The requirement of the chicks for Phe, based on weight gain and feed efficiency, was determined to be 0.39% of the diet. In experiment 2, the IAA - Phe (10% of the diet) or excess Phe (2% of the diet) was added to a diet containing 0.44% Phe. Chicks given the IAA - Phe or excess Phe had significantly slower growth rates than chicks given the basal diet (P > or = 0.05). The activities of the major hepatic enzyme of Phe catabolism, Phe hydroxylase (PAH), were significantly higher than that of chicks fed the basal diet when the chicks were fed the diets containing IAA - Phe plus 1.1% Phe (P > or = 0.05) but not when chicks were fed the diet containing IAA - Phe alone. The activity of PAH in chicks given the excess (2%) Phe was nearly 4 times the activity of PAH in chicks given the basal diet. Adding IAA - Phe to the diet containing excess Phe also resulted in higher PAH activity than was observed in chicks fed the basal diet, although the activity was significantly lower than observed for chicks receiving the diet containing excess Phe alone (P > or = 0.05). It is concluded that hepatic PAH activity in chicks increases primarily in response to its substrate, Phe. A dietary amino acid load without Phe reduces this response to excess Phe.

The use of low-protein, low-phosphorus, amino acid- and phytase-supplemented diets on laying hen performance and nitrogen and phosphorus excretion.

Two experiments were conducted to determine whether, by using a low-protein, amino acid-supplemented diet or a low-protein, amino acid-supplemented diet in conjunction with low-P, phytase-supplemented diet, the excretion of N and P could be reduced without affecting the productive performance of laying hens. Eight dietary treatments were assigned to Babcock B300 hens in each of 2 experiments that involved a positive control (16 to 16.5% CP) and a negative control (13% CP) with and without supplementation with the limiting essential amino acids. In experiment 1, supplementing the negative control with lysine, methionine, and tryptophan resulted in performance comparable to that obtained with the positive control, with the exception that egg weight was heavier for the negative control supplemented with amino acids. Supplementing the negative control with additional essential amino acids improved the performance higher than the positive control indicating that the positive control was deficient in one or more essential amino acids. In experiment 2, supplementing the negative control containing 0.2% nonphytate P (NPP) with all the limiting amino acids plus phytase resulted in performance comparable to the positive control group, which was fed 0.4% NPP without phytase. The results of a digestibility assay indicated that daily total P and N excretions of the negative control containing 0.2% NPP and supplemented with limiting amino acids and phytase were reduced by 48 and 45% of the positive control group, respectively, without compromising laying performance.

Dietary arginine intake alters avian leukocyte population distribution during infectious bronchitis challenge.

Although dietary arginine is a factor in immune function and disease resistance, the full range of effects has yet to be described. In this study, the effects of dietary arginine on leukocyte population changes were examined in the peripheral blood and the respiratory tract of chickens inoculated with infectious bronchitis virus (IBV) strain M41. At 2 wk of age, female line P2a White Leghorn-type chickens were randomly assigned to one of three diets with different arginine levels: a marginally deficient diet (0.5%), an adequate diet (1.0%), and a diet containing a high level of arginine (3.0%). All birds were inoculated with IBV at 4 wk of age, and then the peripheral blood and the respiratory lavage were collected at 1 and 7 d postinfection (DPI). The growth rate of birds that received 0.5% arginine was significantly lower than that of birds receiving 1.0 or 3.0% arginine, whereas the growth of the latter groups did not differ. The percentage and absolute number of heterophil (H) and the H/lymphocyte (L) ratio in the peripheral blood at 1 DPI significantly increased as dietary arginine increased. In the respiratory lavage at 1 DPI, the percentage of H also increased with dietary arginine increase. At 7 DPI, the percentage of CD8+ cells from birds fed the deficient diet was lower than those from birds fed the adequate diet and the diet containing a high level of arginine, whereas the cell surface density of CD8 antigen did not vary among groups. These results show that dietary arginine influences the character of the chicken cellular response to IBV and the distribution of responding leukocyte subpopulations in a target tissue for the infection.

The effect of dietary protein level on threonine dehydrogenase activity in chickens.

An experiment was carried out to determine the effect of dietary protein level on the specific activity of hepatic L-threonine dehydrogenase in young growing chicks. Six replicate pens of seven Leghorn chicks were fed semipurified diets containing 23, 27, or 32% CP with identical relative proportions of amino acids in each protein group. Body weights and feed consumption were measured for 3 d, and hepatic mitochondria were isolated for assay of threonine dehydrogenase (TDH) activity. Weight gains and feed efficiency increased at each level of protein supplementation, but feed consumption was not affected by protein level. The specific activity of threonine dehydrogenase in isolated liver mitochondria was significantly (P < 0.05) higher in the 32% CP group than in the 23% CP group, and the activity in the 27% CP group was intermediate. We conclude that moderate increases in dietary protein level result in elevated hepatic threonine dehydrogenase activity in growing chicks.

Arginine-genotype interactions and immune status.

The effects of arginine on selective immune responses were investigated in a high arginine-requiring (HA) and low arginine-requiring (LA) strain of chickens. Female chickens from these strains were fed diet containing a nutritionally inadequate level of arginine (0.53% arginine diet) or a surfeit level of arginine (1.53% arginine diet) for 2 weeks. Compared to LA chickens, HA chickens showed a higher feed efficiency, body weight gain, and relative thymus and spleen weights with L-arginine supplementation (p < 0.05). In both HA and LA chickens, a deficiency of arginine significantly decreased the delayed-type hypersensitivity response (p < 0.05) and nitric oxide (NO) production from macrophages. Chickens of the HA strain had higher NO production than those of LA strain with E. coli lipopolysaccharide (LPS) activation. This study indicates that dietary arginine concentration influences the immune status of chickens and that strains that differ in arginine requirements for growth may differ in their arginine needs for immune function.

Characterization of hepatic L-threonine dehydrogenase of chicken.

The L-threonine dehydrogenase (TDH) was purified approximately 1300-fold to a specific activity of approximately 18000 unit mg(-1) from chicken (Gallus domesticus) liver mitochondria. Purification was obtained by sequential chromatography on DEAE Cellulose, Phenyl Sepharose High Performance hydrophobic interaction, Affi-Gel Blue affinity and Matrex Gel Red A columns. The molecular weight of the subunit was estimated to be 36 kDa by sodium dodecyl-polyacrylamide gel electrophoresis. An apparent molecular mass of native protein between 62 and 74 kDa was obtained by gel filtration chromatography, suggesting a dimeric structure of TDH. The isoelectric point of TDH was determined by isoelectric focusing to be 5.3. Partial amino-terminal sequence analyses, carried out on two purified preparations of TDH, revealed a high degree of homology to the reported sequence of porcine TDH. The Michaelis constants for L-threonine and NAD for partially purified chicken hepatic TDH are 5.38 and 0.19 mM, respectively.

Dietary supplements of mixtures of indispensable amino acids lacking threonine, phenylalanine or histidine increase the activity of hepatic threonine dehydrogenase, phenylalanine hydroxylase or histidase, respectively, and prevent growth depressions in chicks caused by dietary excesses of threonine, phenylalanine, or histidine*

Experiments were carried out to determine whether the addition of a mixture of indispensable amino acids (IAA) lacking in threonine, phenylalanine or histidine, respectively, to a nutritionally complete diet would increase the hepatic activities of the rate-limiting enzymes for catabolism of threonine, phenylalanine or histidine and prevent the adverse effects of the amino acid on growth when the dietary level of the amino acid is excessive. Week old Leghorn chicks were fed semi-purified diets containing 19% crude protein to which were added no IAA supplement or 10% crude protein from an IAA mix and 5 graded levels of either L-threonine, L-phenylalanine or L-histidine in a 2 x 5 factorial arrangement of treatments. Each amino acid was investigated in a separate experiment involving four replicate pens (seven chicks each) per diet. Weight gains and feed consumptions were determined on the fourteenth day of each experiment. The groups receiving no excess, and 1.0% or 2.0% excesses of amino acids were sampled on the fifteenth day for enzyme activities and plasma amino acid concentrations. Weight gain and/or feed consumption were lower, and plasma concentrations of threonine, phenylalanine and histidine were higher, in chicks receiving 1.5 to 2.0% dietary additions of threonine, phenylalanine, and histidine, respectively, than in chicks that did not receive these amino acids. Chicks that received the amino acids in diets that also contained the IAA supplement had better growth and feed consumption, lower plasma concentrations of threonine, phenylalanine or histidine, higher plasma concentrations of other indispensable amino acids, and higher activities of threonine dehydrogenase, phenylalanine hydroxylase, and histidase than chicks receiving excess amino acids in the absence of IAA supplements. We conclude that the dietary level of protein, not the dietary level of individual amino acids, is the primary determinant of the activity of amino acid degrading enzymes in liver. The increased activity of these enzymes may be the mechanism by which dietary protein alleviates the adverse effects of excessive levels of individual amino acids.

Lysine and arginine requirements of broiler chickens at two- to three-week intervals to eight weeks of age.

Four experiments were conducted to determine the arginine and lysine requirements of male chickens for 2- to 3-wk intervals from the time of hatching until 8 wk of age. Weight gain, breast muscle growth, and feed efficiency were used as response for each interval. Dietary requirements for lysine and arginine were estimated by broken-line regression analysis of responses to six or seven dietary levels of each amino acid. Dietary crude protein levels were 22, 21, 20, and 18% in four consecutive experiments from 0 to 2, 2 to 4, 3 to 6, and 5 to 8 wk of age. An occasional estimate of requirement was not determined (ND) because the response did not conform to the regression model. The values for lysine and arginine requirements determined from breast muscle gain (weight gain of pectoralis major plus pectoralis minor) were not significantly higher than those from body weight gain. However, they tended to be higher than for feed efficiency for 0-to-2 and 2-to-4-wk-old broilers. Lysine and arginine requirements, as percentages of total amino acid in the diet, for maximum breast muscle growth were, respectively, 1.32+/-0.01% and 1.27+/-0.00% to 2 wk of age, 1.21+/-0.06% and ND for 2 to 4 wk of age, 0.99+/-0.02% and 0.97+/-0.02% for 3 to 6 wk of age, and 0.81+/-0.01% and 0.83+/-0.02% for 5 to 8 wk of age. Calculated digestible lysine and arginine requirements were, respectively, 1.24 and 1.19% to 2 wk of age, 1.11% and ND for 2 to 4 wk of age, 0.92% and 0.91% for 3 to 6 wk of age, and 0.75 and 0.78% for 5 to 8 wk of age. The requirements for lysine and arginine were similar except for the earliest age group for which the lysine requirement appeared to be slightly higher than that of arginine.