Impaired translation initiation activation and reduced protein synthesis in weaned piglets fed a low-protein diet

Weanling mammals (including infants) often experience intestinal dysfunction when fed a high-protein diet. Recent work with the piglet (an animal model for studying human infant nutrition) shows that reducing protein intake can improve gut function during weaning but compromises the provision of essential amino acids (EAA) for muscle growth. The present study was conducted with weaned pigs to test the hypothesis that supplementing deficient EAA (Lys, Met, Thr, Trp, Leu, Ile and Val) to a low-protein diet may maintain the activation of translation initiation factors and adequate protein synthesis in tissues. Pigs were weaned at 21 days of age and fed diets containing 20.7, 16.7 or 12.7% crude protein (CP), with the low-CP diets supplemented with EAA to achieve the levels in the high-CP diet. On Day 14 of the trial, tissue protein synthesis was determined using the phenylalanine flooding dose method. Reducing dietary CP levels decreased protein synthesis in pancreas, liver, kidney and longissimus muscle. A low-CP diet reduced the phosphorylation of eukaryotic initiation factor (eIF) 4E-binding protein-1 (4E-BP1) in skeletal muscle and liver while increasing the formation of an inactive eIF4E.4E-BP1 complex in muscle. Dietary protein deficiency also decreased the phosphorylation of mammalian target of rapamycin (mTOR) and the formation of an active eIF4E.eIF4G complex in liver. These results demonstrate for the first time that chronic feeding of a low-CP diet suppresses protein synthesis in animals partly by inhibiting mTOR signaling. Additionally, our findings indicate that supplementing deficient EAA to low-protein diets is not highly effective in restoring protein synthesis or whole-body growth in piglets. We suggest that conditionally essential amino acids (e.g., glutamine and arginine) may be required to maintain the activation of translation initiation factors and optimal protein synthesis in neonates.     

Amino acids modulates the intestinal proteome associated with immune and stress response in weaning pig

The objective of this study was to investigate the effects of free amino acids supplementation to protein restricted diet on the intestinal morphology and proteome composition in weaning pigs. Weanling piglets were randomly fed one of the three diets including a corn-soybean based control diet and two lower protein diets with or without free amino acids supplementation for 2 weeks. The jejunum samples of piglets were collected for morphology and proteome analysis. Compared with the control diet, the protein restricted diet had a significant lower average daily gain and higher feed conversion rate. Free amino acids supplementation to the protein restricted diet significantly improved average daily gain and higher feed conversion rate, compared with the protein restricted diet. The villous height in pigs fed the protein restricted diet was lower than that of the control and free amino acids diet. Using two-dimensional gel electrophoresis and mass spectrometry, we identified 16 differentially expressed protein spots in the jejunum of the weaning piglet. These proteins were related to stress and immune response, the metabolism of carbohydrates and lipids, and tissue structure. Based on the proteome and ELISA analysis, free amino acids diet significantly down-regulated the jejunal expression of stress protein heat shock 60 kDa protein. Our results indicated that amino acids supplementation to the protein restricted diet could enhance weight gain and feed efficiency in weanling pigs through improving intestinal nutrient absorption and transportation, gut health, and mucosal immunity.     

Supplementation with branched-chain amino acids to a low-protein diet regulates intestinal expression of amino acid and peptide transporters in weanling pigs

This study determined the effects of dietary branched-chain amino acids (AA) (BCAA) on growth performance, expression of jejunal AA and peptide transporters, and the colonic microflora of weanling piglets fed a low-protein (LP) diet. One hundred and eight Large White × Landrace × Duroc piglets (weaned at 28 days of age) were fed a normal protein diet (NP, 20.9 % crude protein), an LP diet (LP, 17.1 % crude protein), or an LP diet supplemented with BCAA (LP + BCAA, 17.9 % crude protein) for 14 days. Dietary protein restriction reduced piglet growth performance and small-intestinal villous height, which were restored by BCAA supplementation to the LP diet to values for the NP diet. Serum concentrations of BCAA were reduced in piglets fed the LP diet while those in piglets fed the LP + BCAA diet were similar to values for the NP group. mRNA levels for Na⁺-neutral AA exchanger-2, cationic AA transporter-1, b^0,+ AA transporter, and 4F2 heavy chain were more abundant in piglets fed the LP + BCAA diet than the LP diet. However, mRNA and protein levels for peptide transporter-1 were lower in piglets fed the LP + BCAA diet as compared to the LP diet. The colonic microflora did not differ among the three groups of pigs. In conclusion, growth performance, intestinal development, and intestinal expression of AA transporters in weanling piglets are enhanced by BCAA supplementation to LP diets. Our findings provide a new molecular basis for further understanding of BCAA as functional AA in animal nutrition.     

Muscle and liver protein metabolism in rats fed raw or heat-treated pea seeds

Raw or extruded pea (Pisum sativum, cv. Ballet) diets with or without supplementary amino acids were fed for 15 days to young growing rats and the effects on tissue weights, liver and muscle protein metabolism and hormone levels monitored. Body weight gain, liver and gastrocnemius muscle weights and protein contents were reduced and some key hormones altered when rats were fed unsupplemented raw pea diets. This appeared to be a result of amino acid deficiencies in the diet, the action of antinutritional factors and the refractory nature of the reserve proteins and other seed components. However, this did not in itself improve the nutritional performance of the rats due to the overriding effects of the amino acid deficiencies in the pea diets. After supplementation, extruded peas supported much higher rates of growth and skeletal muscle deposition than did supplemented raw peas. Despite this, the weight gains remained less than achieved on a high quality control diet. Protein synthesis and degradation rates in skeletal muscles and total protein contents were similar to control values. The lower growth rate did not appear to be due to impaired deposition of skeletal muscle. Deposition of other body components, possibly lipids, may have been lowered by supplemented extruded pea diets. Liver protein levels were reduced in rats fed supplemented raw peas and blood corticosterone was elevated. In conclusion, extrusion treatment of peas in combination with amino acid supplementation appeared to abolish the negative effects of peas on skeletal muscle deposition.     

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。     

Studies on limiting essential amino acids in grieves as source of dietary protein for rainbow trout (oncorhynchus mykiss)

Diets were computed to contain equal concentrations of digestible crude protein either of wheat gluten (diet 1) or of grieves (diets 2–8). Per kg dry diet, 41 g crystalline amino acids were supplemented. All diets contained at least 1.2 g Lys per MJ digestible energy (DE). In diet 2, ratios of Met + Cys, Trp, Leu, Ile and Phe to Lys were about equal to those in diet 1. In each of diets 3–7, one of the respective amino acids, in diet 8 all five were replaced by Glu in the supplemented mixture of amino acids.

Each diet was fed to triplciate groups of 20 trout during a trial lasting 66 days. Trout fed the diet containing wheat gluten consumed more dry matter and showed higher growth rates as well as higher protein contents in their gained body mass than trout fed diets based on grieves. Supplementing Met plus Trp significantly improved dry matter intake, growth rate and protein content of gain, though not to the level of trout fed the wheat gluten diet, whereas Leu, Ile and Phe showed no such effect. When grieves were not supplemented with both Met and Trp, gain in body mass contained significantly more lipids. DE required per kg gain by trout fed wheat gluten, grieves + Met + Trp or grieves without supplementation of Met and Trp was 20.1, 21.2 and 29.9 MJ, respectively.     

Proteomic sensitivity to dietary manipulations in rainbow trout

Changes in dietary protein sources due to substitution of fish meal by other protein sources can have metabolic consequences in farmed fish. A proteomics approach was used to study the protein profiles of livers of rainbow trout that have been fed two diets containing different proportions of plant ingredients. Both diets control (C) and soy (S) contained fish meal and plant ingredients and synthetic amino acids, but diet S had a greater proportion of soybean meal. A feeding trial was performed for 12 weeks at the end of which, growth and protein metabolism parameters were measured. Protein growth rates were not different in fish fed different diets; however, protein consumption and protein synthesis rates were higher in the fish fed the diet S. Fish fed diet S had lower efficiency of retention of synthesised protein. Ammonia excretion was increased as well as the activities of hepatic glutamate dehydrogenase and aspartate amino transferase (ASAT). No differences were found in free amino acid pools in either liver or muscle between diets. Protein extraction followed by high-resolution two-dimensional electrophoresis, coupled with gel image analysis, allowed identification and expression of hundreds of protein. Individual proteins of interest were then subjected to further analysis leading to protein identification by trypsin digest fingerprinting. During this study, approximately 800 liver proteins were analysed for expression pattern, of which 33 were found to be differentially expressed between diets C and S. Seventeen proteins were positively identified after database searching. Proteins were identified from diverse metabolic pathways, demonstrating the complex nature of gene expression responses to dietary manipulation revealed by proteomic characterisation.     

Lupinus luteus,Vicia sativa and Lathyrus cicera as protein sources for piglets: ileal and total tract apparent digestibility of amino acids and antigenic effects

Twenty-four male piglets, weaned at 28 days of age, were used to measure the total and ileal digestibility and serum immune responses to dietary leguminous seeds. The experimental diets consisted of a control starter (C) and three other diets prepared by replacing 30% of the crude protein content of the C diet by the protein of Lupinus luteus (LL), Vicia sativa (VS) or Lathyrus cicera (LC). The total tract apparent digestibility (TTAD) of energy and crude protein (CP) was lowest (P<0.001) for the LC diet. Similarly TTAD of lysine, methionine and threonine were also lowest for the LC diet. The apparent ileal digestibility (AID) of lysine, methionine and threonine were lowest for the LC diet. AID of lysine and methionine was highest for the LL seeds (0.764 and 0.834, respectively) and lowest (P<0.05) for the LC seeds (only 0.454 and 0.420, respectively). Immunoblots of individual pig serum were made to detect both residual antigenic storage proteins of the seeds of each legume used and IgGs specific to those storage proteins. Antibodies against β-conglutin of L. luteus, vicilin of V. sativa and vicilin of L. cicera were detected 28 days after feeding the diet in the sera of piglets fed on the LL, VS or LC diets, respectively. Conversely, no storage protein was found in the serum of any piglet fed either on LL, VS or LC diets. The presence of antibodies against β-conglutin of lupine, vicilin of V. sativa and vicilin of L. cicera, respectively indicated an immune response in weaned piglets. The absence of residual antigenic proteins may be due to the digestive adaptation of the piglet to the legume-based diets. However, no direct relation between the differences in digestibility coefficients among the legume seeds and their antigenicity was established.     

Effect of low-protein amino acid-supplemented diets on the growth performance, gut morphology, organ weights and digesta characteristics of weaned pigs

A 21-day study was conducted to determine whether isoleucine might limit the performance of piglets fed low-crude protein (CP), amino acid (AA)-supplemented diets and to investigate the potential benefits of low-CP diets on gastrointestinal health in weaned pigs. Ninety-six piglets (initial BW = 6.44 ± 0.14 kg), housed four per pen, were randomly assigned to one of four diets, resulting in six replicate pens per diet. Dietary treatments were as follows: (1) 210 g/kg CP diet, (2) 190 g/kg CP diet deficient in isoleucine, (3) 190 g/kg CP diet supplemented with crystalline isoleucine up to the level in the 210 g/kg CP diet and (4) 170 g/kg CP diet supplemented with isoleucine and valine on the ideal protein ratio basis (60% and 70% relative to lysine, respectively). Pigs were allowed to adapt to the new environment for 4 days before the experiment commenced. Overall, pigs fed the 210 g/kg CP diet had higher (P < 0.05) average daily gain and lower (P < 0.05) feed : gain ratio compared with those fed the other diets. The faecal consistency score of pigs fed the 210 g/kg CP diet was higher (P < 0.05) than those fed the other diets. Pigs fed the 170 g/kg diet had lower (P=0.02) small intestine weight than those fed the 210 g/kg CP diet. Pigs fed the 210 g/kg CP diet had deeper (P < 0.05) crypt in the duodenum and ileum and higher (P < 0.05) ammonia N concentration in caecal digesta than those fed the other diets. There were no effects of diet on microbial population and volatile fatty acid concentration in the caecal digesta except for propionic acid whose concentration was higher (P < 0.05) for pigs fed the 170 g/kg diet than those fed the 190+isoleucine and the 210 g/kg CP diets. The results indicate that the low-CP, AA-supplemented diet reduced crypt hypertrophy, ammonia N concentration in the caecal digesta, small intestine weight and the performance of piglets. Also, the results of the current study were inconclusive with respect to whether isoleucine may limit the performance of pigs fed a low-CP, AA-supplemented diet.     

Ileal amino acids digestibility of sorghum in weaned piglets and growing pigs

The objective of the study was to determine the coefficients of ileal apparent digestibility (CIAD) of sorghum protein and amino acids (AA) in weaned piglets and growing pigs. Digestibility coefficients were estimated using the regression and difference methods for the weaned piglets; and the direct and difference methods for the growing pigs. To test the hypothesis that CP and AA digestibility of sorghum is lower in weaned piglets than in growing pigs, two experiments were conducted. In experiment one, 20 weaned piglets were fitted with a 'T' cannula at 21 days of age and were fed for 2 weeks one of five dietary treatments: a reference or control diet providing 200 g of CP/kg from casein (C) as the sole protein source, and four casein-sorghum (C-S) diets kept isoproteic to C by the appropriate adjustment of C and maize starch proportions; the amount of sorghum (S) in these diets was 135, 307, 460 and 614 g/kg. In experiment 2, fifteen castrated pigs weighing 57.8 ± 2.8 kg were used and randomly allotted to one of three dietary treatments: a reference casein-maize starch diet containing C as the sole protein source, a C-S diet, both diets containing 160 g of CP/kg, and a fortified S diet containing 68 g of CP/kg. In piglets the CIAD for CP and AA decreased linearly (P < 0.05) as the amount of S in the diet increased. The average ileal digestibility of AA from C was 0.858 ± 0.111, and decreased to 0.663 ± 0.191 at the higher S level. The CIAD estimated using the regression or difference methods were similar for leucine, cysteine, glutamic acid, serine, alanine and tyrosine, and different for the other AA. In growing pigs the CIAD of protein and AA (except alanine and cysteine) were similar (P > 0.05) for the C and the C-S diets, but higher (P < 0.05) than those for the S diet. The CIAD for S obtained by the difference method were higher (P < 0.05) than those obtained using the direct method, except for lysine, isoleucine, valine, methionine, threonine and cysteine. The results indicate that except for lysine and cysteine, growing pigs' ability to digest AA and protein is superior than weaned piglets.     

A diet defined by its content of bovine milk exosomes and their RNA cargos has moderate effects on gene expression,amino acid profiles and grip strength in skeletal muscle in C57BL/6 mice

Exosomes are nanoparticles that transfer cargos from donor cells to recipient cells where they elicit changes in gene expression and metabolism. Evidence suggests that exosomes and their cargos are also absorbed from dietary sources such as bovine milk, and bovine exosomes promote the growth of myofibers in murine C2C12 myotube cell cultures. The aim of the current study was to determine whether the dietary intake of bovine milk exosomes alters strength, gene expression and amino acid profiles in murine skeletal muscles. Male and female C57BL/6 mice, age three weeks, were fed an AIN93G-based, exosome and RNA-depleted (ERD) diet for six weeks; controls were fed an exosome and RNA-sufficient (ERS) diet. Variables of feeding behavior, metabolism, grip strength, liver and kidney function, amino acid profiles, and gene expression patterns were analyzed by using metabolic cages, grip strength analyzers, clinical chemistry analyzers, targeted LC/MS–MS, and RNA sequencing analysis. The diets had no effect on food and water intake, respiratory exchange rate, physical activity, grip strength, markers of liver and kidney dysfunction, and amino acid profiles in muscle. Only twelve and nine mRNAs were differentially expressed in skeletal muscle from female and male mice, respectively, fed ERD and ERS diets. The modest effect of the ERD diet on gene expression and levels of free amino acids in skeletal muscle is consistent with observations that bovine milk exosomes and their cargos accumulate in tissues other than skeletal muscle.     

Effects of amino acid supplementation on the nutritive quality of fermented linseed meal protein in the diets for rohu, Labeo rohita, fingerlings

A feeding trial was conducted for 8 weeks to examine the effects of partial substitution of fish meal (FM) protein (crude protein content: 58.5%) with linseed meal protein with and without supplemental amino acids in diets for rohu Labeo rohita (Hamilton), fingerlings (mean weight: 1.50 ± 0.3 g). Prior to incorporation into the diets, linseed meal was fermented with lactic acid bacteria ( Lactobacillus acidophilus ) to reduce/eliminate the antinutritional tannin and phytic acid factors. Twelve experimental diets (diets D1–D12) were formulated to replace the FM protein from a reference diet (RD) with linseed meal protein at different levels (four sets of diets, of which each set of three diets contained 25%, 50% and 75% replacement of FM protein by linseed meal protein, respectively). Diets D1–D3 were not supplemented with any amino acid. Lysine was supplemented in diets D4–D6. Diets D7–D9 were supplemented with methionine + cystine (together), and diets D10–D12 contained lysine and methionine + cystine (together). Lysine and methionine + cystine (together) were added to the diets at 5.7% and 3.1% of dietary protein, respectively. The groups of fish fed diets without amino acid supplementation had significantly lower percentages of weight gain, specific growth rate and high feed : gain ratio than the fish groups fed other experimental diets. The addition of lysine and methionine + cystine to the diet in which 50% of the FM protein was replaced by linseed meal protein (diet D11) significantly improved fish performance. The results of the present study suggest that rohu fingerlings can effectively utilize the supplemented amino acids and that linseed meal protein can replace up to 50% of the FM protein in rohu diets if the linseed meal is properly processed (fermented) and supplemented with the lacking amino acids.     

Effects of dietary protein and amino acid levels on the expression of selected cationic amino acid transporters and serum amino acid concentration in growing pigs

The absorption of lysine is facilitated by leucine, but there is no information regarding the effect of crude protein, lysine and leucine levels on the expression of cationic amino acid transporters in pigs. Therefore, an experiment was conducted with 20 pigs (14.9 +/- 0.62 kg initial body weight) to evaluate the effect of two protein levels, and the content of lysine, threonine, methionine and leucine in low crude protein diets on the expression of b(0,+) and CAT-1 mRNA in jejunum, Longissimus dorsi and Semitendinosus muscles and serum concentration of amino acids. Treatments were as follows: (i) wheat-soybean meal diet, 20% crude protein (Control); (ii) wheat diet deficient in lysine, threonine and methionine (Basal diet); (iii) Basal diet plus 0.70% L-lysine, 0.27% L-threonine, 0.10% DL-methionine (Diet LTM); (iv) Diet LTM plus 0.80% L-leucine (Diet LTM + Leu). Despite the Basal diet, all diets were formulated to meet the requirements of lysine, threonine and methionine; Diet LTM + Leu supplied 60% excess of leucine. The addition of lysine, threonine and methionine in Diet LTM increased the expression of b(0,+) in jejunum and CAT-1 in the Semitendinosus and Longissiums muscles and decreased CAT-1 in jejunum; the serum concentration of lysine was also increased (p < 0.01). Further addition of L-leucine (Diet LTM + Leu) decreased the b(0,+) expression in jejunum and CAT-1 in the Longissimus dorsi muscle (p < 0.05), increased the serum concentration ofleucine and arginine and decreased the concentration of isoleucine (p < 0.05). Pigs fed the Control diet expressed less b(0,+) in jejunum, and CAT-1 in the Semitendinosus and Longissiums muscles expressed more CAT-1 in jejunum (p < 0.05) and had lower serum concentration ofisoleucine, leucine and valine (p < 0.05), but higher lysine concentrations (p < 0.01) than those fed Diet LTM. These results indicated that both, the level and the source of dietary amino acids, affect the expression of cationic amino acid transporters in pigs fed wheat-based diets.     

Effects of diet and of alloxan-diabetes on the activity of branched-chain 2-oxo acid dehydrogenase complex and of activator protein in rat tissues.

The total activities (sum of active and inactive forms) of branched-chain 2-oxo acid dehydrogenase complex in tissues of normal rats fed on a standard diet were (unit/g wet wt.): liver, 0.82; kidney, 0.77; heart, 0.57; hindlimb skeletal muscles, 0.034. Total activity was decreased in liver by 9%- or 0%-casein diets and by 48 h starvation, but not by alloxan-diabetes. Total activities were unchanged in kidney and heart. The amount of active form of the complex (in unit/g wet wt. and as % of total) in tissues of normal rats fed on standard diet was: liver, 0.45, 55%; kidney, 0.55, 71%; heart, 0.03, 5%; skeletal muscle less than 0.007, less than 20% (below lower limit of assay). The concentration of the active form of the complex was decreased in liver and kidney, but not in heart, by low-protein diets, 48 h starvation and alloxan-diabetes. In heart muscle alloxan-diabetes increased the concentration of active complex. The concentration of activator protein (which activates phosphorylated complex without dephosphorylation) in liver and kidney was decreased by 70-90% by low-protein diets and 48 h starvation. Alloxan-diabetes decreased activator protein in liver, but not in kidney. Evidence is given that in tissues of rats fed on a normal diet approx. 70% of whole-body active branched chain complex is in the liver and that the major change in activity occasioned by low-protein diets is also in the liver.     

Effect of a Low-Protein Diet Supplemented with Ketoacids on Skeletal Muscle Atrophy and Autophagy in Rats with Type 2 Diabetic Nephropathy

A low-protein diet supplemented with ketoacids maintains nutritional status in patients with diabetic nephropathy. The activation of autophagy has been shown in the skeletal muscle of diabetic and uremic rats. This study aimed to determine whether a low-protein diet supplemented with ketoacids improves muscle atrophy and decreases the increased autophagy observed in rats with type 2 diabetic nephropathy. In this study, 24-week-old Goto-Kakizaki male rats were randomly divided into groups that received either a normal protein diet (NPD group), a low-protein diet (LPD group) or a low-protein diet supplemented with ketoacids (LPD+KA group) for 24 weeks. Age- and weight-matched Wistar rats served as control animals and received a normal protein diet (control group). We found that protein restriction attenuated proteinuria and decreased blood urea nitrogen and serum creatinine levels. Compared with the NPD and LPD groups, the LPD+KA group showed a delay in body weight loss, an attenuation in soleus muscle mass loss and a decrease of the mean cross-sectional area of soleus muscle fibers. The mRNA and protein expression of autophagy-related genes, such as Beclin-1, LC3B, Bnip3, p62 and Cathepsin L, were increased in the soleus muscle of GK rats fed with NPD compared to Wistar rats. Importantly, LPD resulted in a slight reduction in the expression of autophagy-related genes; however, these differences were not statistically significant. In addition, LPD+KA abolished the upregulation of autophagy-related gene expression. Furthermore, the activation of autophagy in the NPD and LPD groups was confirmed by the appearance of autophagosomes or autolysosomes using electron microscopy, when compared with the Control and LPD+KA groups. Our results showed that LPD+KA abolished the activation of autophagy in skeletal muscle and decreased muscle loss in rats with type 2 diabetic nephropathy.     

the effect of the combination of microbial phytase and amino acid supplementation of diets for finishing pigs on p and n excretion and carcass quality

The objective of this study was to evaluate the effect of a combined low-protein, low-phosphorus diet supplemented with limiting amino acids and microbial phytase on performance, nutrient utilization and carcass characteristics of late-finishing barrows. 4?×?8 crossbreed barrows were continuously housed in metabolism cages from 70?–?110?kg BW and were fed diets, either conventional (A) or protein reduced (B) or protein and phosphorus reduced diets (C) based on barley, maize and soybean meal. Diet A (positive control) contained in air dry matter 13% and 10% CP as well as 0.49% and 0.42% P at growth phases I (70?–?100?kg BW) or II (100?–?110?kg BW), respectively. Diet B was low in CP (11.3%, 8.4%), diet C low in CP and low in P (CP: as B, P: 0.36%, 0.30%). To diet B the limiting amino acids lysine, methionine, threonine and trypthophan were added to meet the levels in diet A. To diet C the limiting amino acids and 800 FTU/kg Aspergillus-phytase were supplemented. At the end of the balance periods the barrows were slaughtered, the carcasses scored and loin chops, ham and Phalanx prima IV were analysed for nutrients and minerals. The CP or P reduction in diets B and C did not generally negatively affect growth, feed efficiency, absolute nitrogen retention or overall carcass performances of the pigs. With the low CP diets B and C, N excretion per unit BWG was decreased by about 23%. The addition of microbial phytase (diet C) increased apparent total tract digestibility of P by about 20%. In spite of 30% reduction of P intake (diet C), the absolute P retention related to 1?kg BW did not differ between treatments. Thus, phytase supplementation in diet C reduced P excretion per unit BWG by about 33%. Phytase raised apparent digestibility of Zn by about 20% but not Ca digestibility. Generally the carcass traits and meat characteristics were not affected by any of the diet strategies. Mineralization of the Phalanx prima IV was also similar in all treatment groups. However, phytase supplementation led to significantly increased zinc concentration in bones (25%). In contrast, Fe incorporation into the Phalanx prima IV was not affected. In general, the feeding regimen introduced in this experiment offers substantial benefits in maintaining a sustainable environmental-friendly pork production even at the stage of late-finishing barrows.     

Supplementing l-leucine to a low-protein diet increases tissue protein synthesis in weanling pigs

Recent work with young pigs shows that reducing dietary protein intake can improve gut function after weaning but results in inadequate provision of essential amino acids for muscle growth. Because acute administration of l-leucine stimulates protein synthesis in piglet muscle, the present study tested the hypothesis that supplementing l-leucine to a low-protein diet may maintain the activation of translation initiation factors and adequate protein synthesis in multiple organs of post-weaning pigs. Eighteen 21-day pigs (Duroc × Landrace × Yorkshire) were fed low-protein diets (16.9% crude protein) supplemented with 0, 0.27 or 0.55% l-leucine (total leucine contents in the diets being 1.34, 1.61 or 1.88%, respectively). At 35 days of age, protein synthesis was determined using the [²H] phenylalanine flooding-dose technique. Additionally, total and phosphorylated levels of mammalian target of rapamycin (mTOR), ribosomal protein S6 kinase 1 (S6K1), and eIF4E-binding protein-1 (4E-BP1) were measured in longissimus muscle and liver. Compared with the control group, dietary supplementation with 0.55% l-leucine for 2 weeks increased (P < 0.05): (1) the phosphorylated levels of S6K1 and 4E-BP1; (2) protein synthesis in skeletal muscle, liver, the heart, kidney, pancreas, spleen, and stomach; and (3) daily weight gain by 61%. Dietary supplementation with 0.27% l-leucine enhanced (P < 0.05) protein synthesis in proximal small intestine, kidney and pancreas. These novel findings provide a molecular basis for designing effective nutritional means to increase the efficiency of nutrient utilization for protein accretion in neonates.     

Dietary protein intake and skeletal-muscle protein metabolism in rats. Studies with salt-washed ribosomes and transfer factors

1. Aspects of skeletal muscle protein synthesis in vitro were studied in young rats given a low-protein diet for up to 10 days and during re-feeding with an adequate diet. 2. Partially purified muscle transfer factors (transferases I and II), crude and purified (NH(4)Cl-washed) ribosomes and a pH5 enzyme fraction were prepared for this purpose. 3. A marked decrease in the capacity of crude ribosomes to carry out cell-free polypeptide synthesis occurred within 4 days of feeding the low-protein diet. 4. The capacity of salt-washed ribosomes to promote amino acid polymerization, in the presence of added transfer factors and aminoacyl-tRNA, was only slightly decreased by the dietary treatment. 5. However, the capacity of salt-washed ribosomes to bind (14)C-labelled aminoacyl-tRNA was decreased by feeding the low-protein diet. 6. The capacity of the pH5 enzyme fraction to promote amino acid incorporation in a complete cell-free system was decreased within 2 days of feeding the low-protein diet. There is no evidence that the change is associated with aminoacyl-tRNA synthetase or binding enzyme activities of the pH5 fractions. 7. These changes are discussed in relation to the diminished rate of protein synthesis in the intact muscle cell when rats are given a low-protein diet.     

Pre- and postnatal protein undernutrition increases hepatic carnitine palmitoyltransferase I activity and decreases enzyme sensitivity to inhibitors in the suckling rat.

Rats were pair-fed isocaloric diets containing either 25% (control diet) or 6% protein (low-protein diet) during the 5 weeks prior to conception and through the gestation and lactation periods; then, carnitine palmitoyltransferase I (CPT-I) activity was determined in liver and skeletal muscle mitochondria isolated from the corresponding pups. Maternal protein undernutrition increased the activity of hepatic CPT-I all along the suckling period, whereas the activity of the skeletal muscle enzyme was unaffected. Moreover, the sensitivity of hepatic CPT-I to inhibition by both malonyl-CoA and 4-hydroxyphenylglyoxylate was decreased in the low-protein group. These alterations in the properties of hepatic CPT-I may be involved in the appearance of hyperketonemia in the rat pup upon maternal administration of low-protein diets.     

Increased muscle proteasome activities in rats fed a polyunsaturated fatty acid supplemented diet

Changes in the proteasome system, a dominant actor in protein degradation in eukaryotic cells, have been documented in a large number of physiological and pathological conditions. We investigated the influence of monounsaturated or polyunsaturated fatty acids (PUFAs) supplemented diets on the proteasome system, in rat skeletal muscles. Thirty rats were randomly assigned to three groups. The control group received only a standard diet. The monounsaturated fatty acid (MUFA) enriched diet group was fed with 3% sunflower oil in addition to standard food, and the polyunsaturated fatty acid supplemented diet group received 9% Maxepa) in addition to the standard diet. We analyzed muscle proteasome activities and content. Monounsaturated or PUFAs supplemented diets given for 8 weeks induced a significant increase in proteasome activities. With the polyunsaturated fatty acid enriched diet, the chymotrypsin-like and peptidylglutamylpeptide hydrolase activities increased by 45% in soleus and extensor digitorum longus (EDL), and by 90% in the gastrocnemius medialis (GM) muscle. Trypsin-like activity of the proteasome increased by 250% in soleus, EDL and GM. This increase in proteasome activities was associated with a concomitant enhancement in the muscle content of proteasome. Proteasome activities and level were less stimulated with a monounsaturated fatty acid supplemented diet. This study provides evidence that a monounsaturated or polyunsaturated fatty acid supplemented diet may regulate muscle proteasomes. Unsaturated fatty acids are particularly prone to free radical attack. Thus, we suggest that alterations in muscle proteasome may result from monounsaturated and polyunsaturated fatty acid-induced peroxidation, in order to eliminate damaged proteins.