Metabolic Effects of Arginine and Methionine Supplement on Rats Fed Excess Glycine Diets

The contents of glycogen, lipid, urea and amino acids, and some enzyme activities in plasma, liver muscle and urine were determined with rats fed 10 to 12 g of 100 g body weight per day of the 10% casein diet (control) and 10% casein diets containing 7% glycine with or without 1.4% l-arginine HC1 and l-methionine for 7 days.

Nine hours after the final feeding, the amount of liver glycogen was high in the order of rats fed 10% casein diet containing 7% glycine, 10% casein diet containing 7% glycine with l-arginine and l-methionine, and the control. The amount of muscle glycogen was decreased only in those fed the control diet. The amount of liver lipid was increased by the addition of l-arginine and l-methionine to the excess glycine diet. Plasma and urinary urea was increased in animals given the excess glycine diets with or without both amino acids. In plasma liver, and muscle of animals given either of both the excess glycine diets 3 and 9 hr after the feeding, in general, glycine and serine were increased, and threonine and alanine were decreased as compared with those of rats given the control diet. However, the increase of glycine in plasma, liver and muscle detected at 9 hr after feeding the excess glycine diet was slightly prevented by the supplementation of both amino acids to the excess glycine diet. The activities of liver glycine oxidase and ornithine δ-aminotransferase of rats given the excess glycine diet with both amino acids were higher than those of other dietary groups. Liver serine dehydratase and glutamate-oxalacetate transaminase activities were high in the order of the animals fed the control, the excess glycine diet and the excess glycine diet containing both amino acids. Glutamate-pyruvate transaminase activity in the liver of rats fed the excess glycine diets with or without both amino acids were markedly higher than that of those fed the control. The activities of phosphopyruvate carboxylase and aconitase in the liver of animals given the excess glycine diet were higher than those of other dietary groups. Liver pyruvate kinase and glutamate dehydrogenase activities were similar among those dietary groups.     

Metabolism of l-Leucine-U-14C in Young Rats Fed Excess Glycine Diets

A proteinaceous inhibitor of auxin-induced ethylene synthesis by mungbean hypocotyls was purified from mungbean seeds by chromatography on CM-Sephadex C–50 and gel filtration on Sephadex G–200. The molecular weight was estimated to be about 89,000 by gel filtration on Sephadex G–200. The inhibitory protein consisted of two subunits with molecular weights of about 58,000 and 31,000.     

Changes of the Serum Cytokine Contents in Broilers Fed on Diets Supplemented with Nickel Chloride

Cytokines are immunoregulatory proteins which play an important role in the immune system. The purpose of this study was to examine the serum cytokine contents including interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6), interleukin-10 (IL-10), interferon gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) induced by dietary nickel chloride in broilers by enzyme-linked immunospecific assay. A total of 240 one-day-old avian broilers were divided into four groups and fed on a corn–soybean basal diet as control diet or the same basal diet supplemented with 300, 600, and 900 mg/kg of nickel chloride. During the experimental period of 42 days, the results showed that the serum IL-2, IL-4, IL-6, IL-10, IFN-γ and TNF-α contents were lower (p?<?0.05 or p?<?0.01) in the 300, 600, 900 mg/kg groups than those in the control group. It was concluded that dietary nickel chloride in the range of 300 to 900 mg/kg could reduce the serum cytokine contents, which could finally impact the immune function in broilers.     

Changes in Plasma and Tissue Free Amino Acids of Rats Fed Excess Glycine Diets

In order to discover more potent pyrethrum synergists, dihydrodillapiole and furapiole, two dillapiole-based compounds having considerable synergistic activity, were converted into various acyl and ?,β-unsaturated carbonyl derivatives. The synthesis, factor of synergism, spectroscopic data and structure activity relationships of these derivatives are reported.     

Dietary Excess Vanadium Induces Lesions and Changes of Cell Cycle of Spleen in Broilers

The purpose of this 42-day study was to investigate the effects of dietary excess vanadium on spleen growth and lesions by determining morphological changes and cell cycle of spleen. Four hundred twenty 1-day-old avian broilers were divided into six groups and fed on a corn–soybean basal diet as control diet or the same diet amended to contain 5, 15, 30, 45, 60 ppm of vanadium supplied as ammonium metavanadate. When compared with that of control group, the relative weight of spleen was significantly raised in 5- and 15-ppm groups, but depressed in 45- and 60-ppm groups. The gross lesions of spleen showed obvious atrophy with decreased volume and pale color in 45- and 60-ppm groups. Histopathologically, lymphocytes in splenic corpuscle and periarterial lymphatic sheath were variously decreased in number in 30-, 45-, and 60-ppm groups. The percentage of static phase (G₀/G₁) was significantly decreased, and the percentage of synthesis period (S) phase and the proliferating index (PI) were significantly increased in 5- and 15-ppm groups. The percentage of G₀/G₁ phase was significantly increased, and the percentage of mitotic phase (G₂ + M), S phase, and PI significantly decreased in 45- and 60-ppm groups. These results suggested that dietary excess vanadium (45 and 60 ppm) could inhibit growth of spleen and induce lesions in spleen in chicken.     

Changes in Liver Enzyme Activities of Glycine Metabolism of Rats Fed Excess Glycine Diets

A gram negative bacterium isolated from soil was found to produce a high level of endo-β-N-acetylglucosaminidase in the culture medium. The organism was identified as a Flavobacterium sp. from various bacteriological characteristics. The enzyme from the Flavobacterium sp. was purified to homogeneity from culture broth by fractionation with ammonium sulfate and column chromatographies on DEAE-cellulose, hydroxylapatite, and Sephadex G-150 and G-100. The molecular weight of the enzyme was estimated to be 27,000 and 30,000 by gel filtration and SDS-polyacrylamide gel electrophoresis, respectively, and it appeared to consist of a single polypeptide chain. The optimal pH for activity was 5.0 to 6.0 and the stable pH range was 5~7. The Michaelis constant was 0.30 mm with dansyl-Asn-(GlcNAc)₂(Man)₆ as the substrate. The enzyme hydrolyzed oligosaccharides of native ovalbumin, bovine pancreatic ribonuclease B and a yeast invertase.     

The Association Between Cytokines and Intestinal Mucosal Immunity Among Broilers Fed on Diets Supplemented with Fluorine

Fluorine (F) bioaccumulation has been reported in the organs and tissues of organisms, including intestine. The intestinal mucosa is very important to the immune development. Meanwhile, cytokines are present in the normal intestinal mucosal and play an important role in the immune function. Thus, changes of the cytokine contents are related to the state of intestinal mucosal immunity. In this study, we investigated the changes in contents of cytokines such as interleukin-2 (IL-2), interleukin-4 (IL-4), interleukin-6 (IL-6), interferon gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) induced by dietary high F in the mucosa of different parts of intestines (duodenum, jejunum, and ileum) by enzyme-linked immunosorbent assay. A total of 280 one-day-old healthy avian broilers were randomly divided into four groups and fed on a corn–soybean basal diet as control diet (F 22.6 mg/kg) or the same basal diet supplemented with 400, 800, and 1,200 mg?F/kg (high F groups I, II, and III) in the form of sodium fluoride for 42 days. The experimental data showed that the contents of IL-2, IL-4, IL-6, IFN-γ, and TNF-α in the intestinal mucosa were significantly decreased in the high F groups II and III when compared with those of the control group from 14 to 42 days of age. It was concluded that dietary F in the range of 800–1,200 mg/kg significantly reduced the contents of aforementioned cytokines in the intestinal mucosa of broilers, which could impact the function of intestinal mucosal immunity through the pathways that decreased the lymphocyte population and/or lymphocyte activation.     

Increased Apoptotic Lymphocyte Population in the Spleen of Young Chickens Fed on Diets High in Molybdenum

The experiment was conducted with the objective of examining the effects of high molybdenum (Mo) on the apoptosis of splenic lymphocytes in broilers by the methods of experimental pathology and flow cytometry (FCM). Three hundred 1-day-old avian broilers were divided into four groups of 75 each and were fed on control diet (Mo 13 mg/kg) and high Mo diets (Mo 500 mg/kg, high Mo group I; Mo 1,000 mg/kg, high Mo group II; and Mo 1,500 mg/kg, high Mo group III) for 42 days. The results showed that the relative weight of spleen was decreased, and lymphocytes were histopathologically decreased in high Mo groups II and III. Ultrastructurally, the apoptotic cells showed typical condensed nuclei with dark-round, petal-like, and horseshoe-like shapes. The mitochondria were swelled, and the endoplasmic reticulum and the Golgi apparatus were dilated in high Mo groups II and III. The statistical analyses by FCM indicated that the percentage of cellular apoptosis was higher in high Mo groups II and III than in control group. Also, the TUNEL staining was consistent with the results of FCM. It was concluded that dietary molybdenum in 1,000 and 1,500 mg/kg caused splenic lesions and lymphocyte apoptosis, which could inhibit the development of spleen and could impair immune function in broilers.     

Changes of IgA+ Cells and Cytokines in the Cecal Tonsil of Broilers Fed on Diets Supplemented with Vanadium

The cecal tonsil of broiler is known as a secondary lymphoid tissue, which is involved in antigen-specific humoral immune responses. The purpose of this study was to investigate the effects of dietary vanadium on the tissue distribution and quantity of immunoglobulin A-positive (IgA(+)) cell in the cecal tonsil by immunohistochemistry. Simultaneously, the changes in interleukin-6 (IL-6), interleukin-10 (IL-10), interferon gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) contents in the cecal tonsil were also quantified by enzyme-linked immunosorbent assay (ELISA). A total of 420 one-day-old avian broilers were divided into six groups and fed on a corn-soybean basal diet (control diet) or the same diet supplemented respectively with 5, 15, 30, 45, and 60 mg/kg of vanadium in the form of ammonium metavanadate for 42 days. The results showed that the population of the IgA(+) cells in the cecal tonsil were significantly lower (p < 0.05 or p < 0.01) in the 45 and 60 mg/kg groups than that in the control group. Meanwhile, IL-10, IFN-γ and TNF-α contents in the cecal tonsil were significantly decreased (p < 0.05 or p < 0.01) in the 30, 45 and 60 mg/kg groups in comparison with those of the control group. However, IL-6 content in the cecal tonsil was only decreased (p < 0.05 or p < 0.01) in 60 mg/kg at 14 and 28 days of age. In conclusion, dietary vanadium in excess of 30 mg/kg reduced the numbers of the IgA(+) cells and changed the contents of the abovementioned cytokines in the cecal tonsil, which may finally impact the function of local mucosal humoral immunity in broilers.     

Excess Dietary Vanadium Induces the Changes of Subsets and Proliferation of Splenic T Cells in Broilers

The purpose of this 42-day study was to investigate the effects of dietary excess vanadium on immune function by determining changes of the subsets and proliferation function of splenic T cells. Four hundred twenty 1-day-old avian broilers were divided into six groups and fed on a corn–soybean basal diet as control diet or the same diet amended to contain 5, 15, 30, 45, and 60 ppm of vanadium supplied as ammonium metavanadate. When compared with those of the control group, the percentage of CD3⁺, CD3⁺CD4⁺, and CD3⁺CD8⁺ of splenic T cells were decreased in the 45 and 60 ppm groups; however, the percentage of CD3⁺ and CD3⁺CD4⁺ were increased in the 5 ppm group, and the CD₄⁺/CD₈⁺ ratios were raised in the 5 and 15 ppm groups at 14 days of age. Meanwhile, the proliferation of splenic T cells were depressed in the 45 and 60 ppm groups but raised in the 5 and 15 ppm groups. Also, the serum interleukin-2 (IL-2) and interleukin-6 (IL-6) contents were decreased in the 45 and 60 ppm groups and increased in the 5 ppm group. It was concluded that dietary vanadium in excess of 30 ppm changed the percentages of splenic T cell subsets and inhibited the proliferation of splenic T cells and reduced the serum IL-2 and IL-6 contents. The cellular immune function was finally impaired in broilers.     

Glycine-U-14C Metabolism in Young Rats Fed the 10% Casein Diets Containing Excess Glycine

Nine hours after rats fed ad libitum for 14 days a 10% caein diet (10C), a 10% casein diet containing 7% glycine (10C7G) and a 10% casein diet containing 7% glycine with 1.4% l-arginine HCI and 0.9% l-methionine (10C7GArgMet) were force-fed 10 ml of each diet suspension containing 5 μCi of glycine-U-¹⁴C per 100 g of body weight, the radioactivity recoveries of ¹⁴C in expired CO₂, tissue components and urine were determined.

The radioactivity recovery of ¹⁴C in the expired C0₂ of the 10C7G group was generally higher than that of the 10C7GArgMet group, and those of both groups would have been much higher than that of the 10C group unless the isotope had been diluted. The amount of expiratory ¹⁴C of rats fed a 25 % casein diet containing 7% glycine was not different from that of the 10C7G group. The recovery of ¹⁴C in the trichloroacetic acid (TCA) soluble fraction of muscle of the 10C7G and the 10C7GArgMet groups were greater than that of the 10C group, but there was no difference between the 10C7G and the 10C7GArgMet groups. The recoveries of ¹⁴C in the TCA soluble fraction and protein of plasma and liver, and the muscle protein were negligible in all the groups. The amount of glycine-¹⁴C incorporated into the carcass lipids of the 10C7GArgMet group was larger than that of other groups. Those in the carcass lipids of the 10C7G and the 10C7GArgMet groups would have been much higher than that of the 10C group unless the dilution of the isotope had taken place. The recoveries of ¹⁴C in the liver and muscle glycogen, and liver lipids were remarkably small in all the groups. From the above results, it was suggested that the degradation of glycine-¹⁴C to expiratory CO₂ was not accelerated, but the rate of incorporation of the isotope into carcass lipids was increased by the supplementation of l-arginine and l-methionine to the 10C7G diet as compared with that of rats fed the 10C7G diet.     

Distribution of Selenium in Bovine Milk and Selenium Deficiency in Rats Fed Casein-based Diets,Monitored by Lipid Peroxide Level and Glutathione Peroxidase Activity

A major proportion of selenium in bovine milk was found in fluorometric analysis to be associated with the casein fraction, largely in alkali-labile form, and the rest with the whey fraction mostly in free selenite form. This uneven distribution of milk selenium seems to provide an explanation for selenium deficiency in purified caseins. The activity of glutathione peroxidase, a selenoprotein, in the liver of growing male rats fed ad libitum low-selenium diets containing either vitamin-free casein or Torula yeast 0.065 ± 0.012 or 0.015 ± 0.004 μ g Se/g diet, respectively) for 3 weeks decreased to 4 to 6% of that of the control rats fed a commercial stock diet (0.185 ± 0.092 μ g Se/g diet). Selenium status was evaluated by three different parameters for the rats assigned under pair-feeding regimen to those vitamin-free casein-based diets which were supplemented with graded levels of selenium as sodium selenite. The hepatic levels of the thiobarbituric acid-reactive substance, an indication of lipid peroxidation, decreased to control level with selenium supplementation per g diet of 0.1 μ g and over. The hepatic glutathione peroxidase activity reached a plateau above a 0.1 μ g/g diet of selenium supplementation, whereas the erythrocyte enzyme activity increased with increasing levels of supplementary selenium. These results support the notion that semi-purified diets containing vitamin-free casein as a prime protein source would not satisfy the selenium requirement of growing animals unless deliberately supplemented with additional selenium.     

Decreased Percentages of the Peripheral Blood T-Cell Subsets and the Serum IL-2 Contents in Chickens Fed on Diets Excess in Fluorine

Three hundred 1-day-old Avian broilers were divided into four groups and fed on control diet (F 23 ppm) and high-fluorine diets (400 ppm, high-fluorine group I; 800 ppm, high-fluorine group II; 1,200 ppm, high-fluorine group III) for 42 days (n?=?75/group). The percentages of CD ₄ ⁺ and CD ₈ ⁺ T cells were decreased in three high-fluorine groups when compared with those of control group. Meanwhile, the CD ₄ ⁺ /CD ₈ ⁺ ratio were lower in high-fluorine group II at 28 days of age and in high-fluorine group III at 42 days of age than in control group. Also, the serum interleukin-2 (IL-2) contents were decreased in three high-fluorine groups when compared with those of control group. Histopathologically, the thymus became hypocellular in three high-fluorine groups. It was concluded that dietary fluorine excess (400~1,200 ppm) reduced the percentages of T-lymphocyte subsets and the serum IL-2 contents, and cellular immune function could be affected in chickens.     

Effects of Alloxan Diabetes and Hypophysectomy on Growth,and Plasma and Liver Free Amino Acids of Rats Fed Excess Glycine Diets

Three chitinases (EC 3.2.1.14) were purified from yam, Dioscorea opposita THUMB, by fractionation with ammonium sulfate, chromatographies on DEAE-Cellulose and DEAE-Sephadex A-50, chromatofocusing and gel filtration on Bio-Gel P-60. The purified enzymes (E-l, E-2 and E-3) showed single bands on sodium dodecylsulfate polyacrylamide gel electrophoresis, and the molecular weights were estimated to be 33,500. The pIs were 4.05 (E-l), 4.0 (E-2) and 3.8 (E-3). All enzymes were glycoproteins and the neutral sugar contents were 3.6% (E-l), 3.6 (E-2) and 0.9% (E-3). The N-terminal amino acids of E-l and E-3 were the same and determined to be histidine. All enzymes hydrolyzed glycolchitin, but not p-nitrophenyl-2-acetamido-2-deoxy-β-d-glucopyranoside or Micrococcus lysodeikticus cell walls. E-l and E-3 were stable in the pH range of 5 ~ 11, and below 60°C. These enzymes showed two optimum pHs around 3.5 and 8.0 or 8.5 with glycolchitin as substrate.     

Blood Selenium in Naturally Fed Horses and the Effect of Selenium Administration

Blood Se of adult horses was 26.1, 25.8, and 27.0 ng/ml (mean values at 3 farms), where the Se of food was about 20 ng/g dry substance. Experimental adult horses which received about 41 ng Se/g food showed 45.3 ng/ml blood. At low Se intake suckling foals show higher blood Se than mares, but with high Se intake, the opposite will occur. This is reflected in milk Se, which raises but slowly with rise of mare’s blood Se. Se in blood plasma and in blood corpuscles is on the same level. The effect of various dose levels of Se on blood Se was studied: From 1.5 to 6 mg Se/week, blood Se rose rather linearity; 18 and 30 rag Se/week gave but slightly more effect than 6 mg.     

Effect of Dietary Taurine on Endogenous Hypercholesterolemia in Rats Fed on Phenobarbital-containing Diets

The effect of dietary taurine on endogenous hypercholesterolemia induced by a phenobarbital-containing diet was investigated. Supplemented taurine did not affect the concentrations of serum cholesterol, but further potentiated the accumulation of hepatic cholesterol in the hypercholesterolemic state induced by phenobarbital. It is suggested that taurine might amplify the hepatic cholesterogenesis in phenobarbital-induced hypercholesterolemia.     

Cottonseed Oil in Diets for Broilers in the Pre-Starter and Starter Phases

The objective of this study was to evaluate the effect of different levels of crude cottonseed oil in isoenergetic diets, with or without supplementation of ferrous sulfate, on performance variables, relative weight of organs, and blood parameters of broilers, and on the economic viability of diets in the periods from 1 to 7 and 1 to 21 days of age. A total of 600 male birds of the Ross line were distributed in a completely randomized design in a (4×2) factorial arrangement with eight treatments (0, 2, 4, and 6% cottonseed oil with and without ferrous sulfate), and five replicates. The following variables were studied: feed intake, weight gain, feed conversion, weight of organs, blood parameters, and yield of carcass and cuts at 21 days. No effects of the levels of cottonseed oil were found on the performance of animals aged 1 to 7 days, or on the relative weights of the organs. In this same period, the weight gain, and the relative weights of heart, liver, and intestine of the animals that received ferrous sulfate were decreased, and feed conversion was worsened. In the period from 1 to 21 days, weight gain increased linearly with the increase in the levels of cottonseed oil. Blood parameters were not influenced by the diets. Crude cottonseed oil can be utilized in diets for broilers in the periods from 1 to 7 and 1 to 21 days of age at up to 6% of inclusion, and supplementation with ferrous sulfate is unnecessary if the differences in metabolization of the cottonseed oil are considered, with and without, it during the diet formulation process.     

Performance of Growing-Finishing Pigs Fed Diets Based on Normal or Low Viscosity Rye Fed With and Without Enzyme Supplementation

One hundred and forty crossbred pigs (Pig Improvement Canada Ltd, Acme Alberta), weighing an average of 21.5 + 3.6kg, were assigned on the basis of sex, weight and litter to one of five dietary treatments. The control diet was barley-based, while in the remaining four treatments, the grain component consisted of either normal or low viscosity rye fed with or without pentosanase. Digestibility coefficients for dry matter, crude protein and gross energy were significantly ( P = 0.0001) higher for the rye-based diets than the barley based diet. There were no differences in digestibility coefficients between normal and low viscosity diets or between diets supplemented or unsupplemented with pentosanase. Over the entire experimental period (21.5 to 100.7kg), pigs fed the normal viscosity rye gained better ( P = 0.001) and had a higher daily intake ( P = 0.001) than pigs fed the low viscosity rye. Pigs fed the low viscosity rye tended to gain slower than pigs fed barley and with lower intake while pigs fed the normal viscosity rye had similar gains and intake to the pigs fed barley. Enzyme supplementation failed to improve pig performance. Males had higher gains and intake than females ( P < 0.05). Enzyme supplementation, rye viscosity or choice of cereal grain had no effect on any of the carcass traits measured. In conclusion, the results of the present experiment indicate that rye is a good alternative to barley for use in growing-finishing rations and becomes increasing attractive as the age of the pig increases. Breeding efforts directed towards reducing the viscosity of rye are unlikely to be successful in improving the nutritive value of rye for swine.