Dietary levels of protein and free amino acids affect pancreatic proteases activities,amino acids transporters expression and serum amino acid concentrations in starter pigs

The dietary contents of crude protein and free amino acids (AA) may affect the protein digestion and AA absorption in pigs. Trypsin and chymotrypsin activities, AA serum concentrations and expression of AA transporters in the small intestine of pigs fed a low protein, AA‐supplemented (19.2%, LPAA) or a high protein (28.1%, HP), wheat‐soybean meal diet were measured in two 14‐d trials. The LPAA diet contained free L‐Lys, L‐Thr, DL‐Met, L‐Leu, L‐Ile, L‐Val, L‐His, L‐Trp and L‐Phe. All pigs were fed the same amount of feed (890 and 800 g/d for trial 1 and 2 respectively). In trial 1, samples of mucosa (duodenum, jejunum and ileum) and digesta (duodenum and jejunum) were collected from 14 pigs (17.2 ± 0.4 kg); in trial 2, blood samples were collected from 12 pigs (12.7 ± 0.3 kg). The trypsin and chymotrypsin activities in both intestinal segments were higher in pigs fed the HP diet (p < 0.01). Trypsin activity was higher in jejunum than in duodenum regardless the dietary treatment (p < 0.05). Pigs fed the LPAA diet expressed more b^0,+AT in duodenum, B⁰AT1 in ileum (p < 0.05), and tended to express more y⁺LAT1 in duodenum (p = 0.10). In pigs fed the LPAA diet, the expression of b^0,+AT was higher in duodenum than in jejunum and ileum (p < 0.01), but no difference was observed in pigs fed the HP diet. Ileum had the lowest b^0,+AT expression regardless the diet. The serum concentrations of Lys, Thr and Met were higher in LPAA pigs while serum Arg was higher in HP pigs (p < 0.05). Serum concentrations of AA appear to reflect the AA absorption. In conclusion, these data indicate that the dietary protein contents affect the extent of protein digestion and that supplemental free AA may influence the intestinal site of AA release and absorption, which may impact their availability for growth of young pigs.     

Dietary amino acids fed in free form and as protein components do not differently affect postprandial plasma insulin, glucagon, growth hormone and corticosterone responses in rats

This study examined, whether the postprandial fate of dietary amino acids from different amino acid sources is regulated by the responses of insulin, glucagon, corticosterone and growth hormone (GH). Male Wistar rats were cannulated in the vena jugularis and assigned to dietary groups. The diets contained 21% casein or the same amino acids in free form. In the free amino acid diets, methionine level was varied between the groups. The feed was supplied in two distinct meals. In previous experiments it was established that oxidative amino acid losses of the free amino acid diets and protein diets were different. After 3 weeks on those diets, it appeared that the differences in postprandial oxidative losses had been diminished. GH was measured every 12 min, from 144 min before the start of the experimental meal over the following 144 min. Insulin and corticosterone were measured six times from the start of the meal until 270 min after the meal. No differences have been observed between the hormonal responses to both meals at day 5 and at day 26. In conclusion, it has been found that the differences in the oxidative losses between protein and free amino acid meals are not mediated by the combined action of the insulin, glucagon, corticosterone and GH. Postprandial catabolism of amino acids is most probably regulated by substrate induction.     

Changes in tissue free amino acid pools in growing chickens fed thermally treated vetch diets

A three‐day assay was developed to evaluate the effect of autoclaving on protein quality of vetch as an alternative to classical growth methods. Male chickens (n = 10/diet) were given approximately isonitrogenous diets based on raw or autoclaved vetch for 3 days. Samples of plasma, muscle and liver were obtained for free amino acid analysis. Heating vetch depressed growth (11.9 vs. 23.2 g/d; p < 0.05). Plasma methionine and histidine increased (0.05 < p < 0.06), while gluconeogenic amino acids tended to decrease (p < 0.10) after heating. Muscle free amino acids did not change except for a trend to increased methionine (p = 0.06) in birds fed autoclaved vetch. In liver, most essential amino acids, glycine, proline and tyrosine increased markedly with heated vetch diet. Correlations between plasma and muscle free amino acids were poor compared with those between plasma and liver free amino acids. Liver free amino acid pool was more sensitive than muscle or plasma pool to amino acid inflow modifications after vetch heating.     

Reduced dietary protein level influences the free amino acid and gene expression profiles of selected amino acid transceptors in skeletal muscle of growing pigs

This study was conducted to evaluate the effect of reduced dietary protein level on growth performance, muscle mass weight, free amino acids (FAA) and gene expression profile of selected amino acid transceptors in different fibre type of skeletal muscle tissues (longissimus dorsi, psoas major, biceps femoris) of growing pigs. A total of 18 cross‐bred growing pigs (Large White × Landrace × Duroc) with initial body weight (9.57 ± 0.67 kg) were assigned into three dietary treatments: 20% crude protein (CP) diet (normal recommended, NP), 17% CP diet (low protein, LP) and 14% CP diet (very low protein, VLP). The results indicated improved feed‐to‐gain ratio was obtained for pigs fed LP and NP diets (p < 0.01), while the pigs fed VLP diet showed the worst growth performance (p < 0.01). There was no significant difference in the weights of longissimus dorsi and psoas major muscle between LP and NP groups (p > 0.05). Majority of the determined FAA concentration of LP group were greater than or equal to those of NP group in both longissimus dorsi and psoas major muscle (p < 0.01). Further, the mRNA expression levels of sodium‐coupled neutral amino acid transceptor 2, L‐type amino acid transceptor 1 and proton‐assisted amino acid transceptors 2 were higher in skeletal muscle tissue in LP group compared to those of the pigs fed NP or VLP diet. These results suggested that reduced dietary protein level (3 points of percentage less than recommended level) would upregulate the mRNA expression of amino acid transceptors to enhance the absorption of FAA in skeletal muscle of growing pigs. There seems to be a relationship between response of AA transceptors to the dietary protein level in skeletal muscle tissue of different fibre type. To illustrate the underlying mechanisms will be beneficial to animal nutrition.     

Estimation of the optimum standardized ileal digestible total sulfur amino acid to lysine ratio in late finishing gilts fed low protein diets supplemented with crystalline amino acids

A total of 90 gilts were used to investigate the effects of various standard ileal digestible (SID) total sulfur amino acid (TSAA) to lysine (Lys) ratios on the performance and carcass characteristics of late finishing gilts receiving low crude protein (CP) diets supplemented with crystalline amino acids (CAA). Graded levels of crystalline methionine (Met) (0, 0.3, 0.5, 0.8 or 1.1 g/kg) were added to the basal diet to produce diets providing SID TSAA to Lys ratios of 0.48, 0.53, 0.58, 0.63 or 0.68. At the termination of the experiment, 30 gilts (one pig per pen) with an average body weight (BW) of 120 kg were killed to evaluate carcass traits. Increasing the SID TSAA to Lys ratio increased average daily gain (ADG) (linear and quadratic effect, P < 0.05), improved feed conversion ratio (FCR) (linear and quadratic effect, P < 0.05) and decreased serum urea nitrogen (SUN) concentration (linear and quadratic effect, P < 0.05) of finishing gilts. No effects were obtained for carcass traits. The optimum SID TSAA to Lys ratios to maximize ADG as well as to minimize FCR and SUN levels were 0.57, 0.58 and 0.53 using a linear‐break point model and 0.64, 0.62 and 0.61 using a quadratic model.     

Dietary N-carbamylglutamate or L-arginine improves fetal intestinal amino acid profiles during intrauterine growth restriction in undernourished ewes

Our previous studies demonstrated that prenatal in utero growth restriction impairs postnatal intestinal function.Thus,improving postpartal intestinal absorption capacity and growth by manipulating the maternal diet prepartum is of importance.This work was conducted to determine whether supplementation of N-carbamylglutamate(NCG)or rumen-protected L-arginine(RP-Arg)increased fetal intestinal amino acid(AA)profiles in intrauterine growth retardation(IUGR)fetuses.On d 35 of gestation,Hu ewes(n=32)carrying twin fetuses were randomized into 4 groups(8 ewes and 16 fetuses in each group),where diets were as follows:100%of nutrient requirements recommended by National Research Council(NRC,2007)(CON);50%of nutrient requirements recommended by NRC(2007)(RES);RES+RPArg(20 g/d),(RES+ARG);and RES+NCG(5 g/d),(RES+NCG).On d 110 of gestation,both fetal and maternal tissues were collected and weighed.Compared with RES,solute carrier family 1,member 5(SLC1A5)was upregulated(P<0.05)within fetal jejunum,duodenum and ileum when supplementing NCG and RP-Arg.Relative to RES,RP-Arg or NCG supplementation to RES resulted in upregulation(P<0.05)of peptide transporter 1 protein abundance within the fetal ileum.NCG or RP-Arg supplementation to RES also upregulated phosphorylated mechanistic target of rapamycin(pmTOR)-to-mTOR ratio in the fetal ileum induced by IUGR(P<0.05).As a result,during IUGR,supplementation of Arg or NCG affected intestinal AA profiles in the fetus in part through controlling mTOR signal transduction as well as AA and peptide transport.Future studies should be conducted to understand the role(if any)of the placenta on the improvement of growth and AA profiles independent of the fetal intestine.This would help demonstrate the relative contribution of intestinal uptake in fetal life.     

Effect of GABA on oxidative stress in the skeletal muscles and plasma free amino acids in mice fed high‐fat diet

Increased levels of plasma free amino acids (pFAAs) can disturb the blood glucose levels in patients with obesity, diabetes mellitus and metabolic syndrome (MS) and are associated with enhanced protein oxidation. Oxidation of proteins, especially in the muscles, can promote protein degradation and elevate the levels of pFAAs. Gamma‐aminobutyric acid (GABA), a food additive, can reduce high‐fat diet (HFD)‐induced hyperglycaemia; however, the mechanisms remain unclear. The aim of this study was to evaluate the effects of GABA on protein oxidation and pFAAs changes. One hundred male C57BL/6 mice were randomly divided into five groups that were fed with control diet, HFD and HFD supplied with 0.2%, 0.12% and 0.06% GABA in drinking water for 20 weeks respectively. HFD feeding led to muscular oxidative stress, protein oxidation, pFAA disorders, hyperglycaemia and augmented plasma GABA levels. Treatment with GABA restored normally fasting blood glucose level and dose‐dependently inhibited body weight gains, muscular oxidation and protein degradation. While medium and low doses of GABA mitigated HFD‐induced pFAA disorders, the high dose of GABA deteriorated the pFAA disorders. Medium dose of GABA increased the levels of GABA, but high dose of GABA reduced the levels of plasma GABA and increased the activity of succinic semialdehyde dehydrogenase in the liver. Therefore, treatment with GABA mitigated HFD‐induced hyperglycaemia probably by repairing HFD‐induced muscular oxidative stress and pFAA disorders in mice. Our data also suggest that an optimal dose of GABA is crucial for the prevention of excess GABA‐related decrease in the levels of pFAA and GABA as well as obesity.     

Additivity of standardized ileal digestibility of amino acids in mixed diets containing multiple protein sources for growing pigs fed three crude protein levels

This study was conducted to investigate the effect of dietary crude protein (CP) levels of semi‐purified diets on the additivity of values for standardized ileal digestibility (SID) of amino acids (AA) in mixed diets from multiple protein sources for growing pigs. A total of 28 barrows (initial BW, 66.4 ± 1.3 kg) were surgically fitted with simple T‐cannulas at the distal ileum and assigned to a replicated 14 × 4 incomplete Latin square design with 14 diets and 4 periods. The 14 experimental diets consisted of a nitrogen‐free diet; a corn‐based diet (80 g CP/kg); nine semi‐purified diets containing soya bean meal (SBM), canola meal (CM) or corn distillers dried grains with solubles (cDDGS), each type (protein source) of semi‐purified diets supplied 80, 120 or 160 g CP/kg, respectively; three mixed diets based on corn, SBM, CM and cDDGS formulated to contain 120, 160 and 200 g CP/kg respectively. Pigs were fed each of the 14 diets during a seven‐day period, and ileal digesta were collected from 08:00 a.m to 6:00 p.m on day 6 and 7. Chromic oxide was added as an indigestible marker. Results indicated that the SID of CP and AA were not affected by CP levels (p > .05). Values for SID of AA were additive (p > .05) with the exception of His and Lys; Arg and Lys; Arg, Lys, Thr, Asp, Cys and Gly in the mixed diets containing 120, 160 and 200 g CP/kg respectively (p < .05). In conclusion, additivity of SID values of AA in the mixed diets at different CP levels was not affected by the CP levels of semi‐purified diets for growing pigs. Therefore, it is recommended that SID values of AA should be used to formulate practical diets containing multiple ingredients for pigs.     

Branched‐chain amino acid ratios in low‐protein diets regulate the free amino acid profile and the expression of hepatic fatty acid metabolism‐related genes in growing pigs

Liver metabolism is affected by nutrients. The aim of this study was to explore the effects of low‐protein diets (17% crude protein, CP) supplemented with branched‐chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile) and valine (Val), on hepatic amino acid profile and lipid metabolism in growing pigs. The ratio of Leu : Ile : Val in all groups was 1 : 0.51 : 0.63 (20% crude protein, CP), 1 : 1 : 1 (17% CP), 1 : 0.75 : 0.75 (17% CP), 1 : 0.51 : 0.63 (17% CP) and 1 : 0.25 : 0.25 (17% CP) respectively. Results revealed that compared to the positive control group (1 : 0.51 : 0.63, 20% CP), the low‐protein diets significantly augmented the concentrations of most essential amino acids and non‐essential amino acids (p < .05), with the greatest values observed in the 1 : 0.25 : 0.25 group. Moreover, relative to the control, the low‐protein diets with the Leu : Ile : Val ratio ranging from 1 : 0.75 : 0.75 to 1 : 0.25 : 0.25 markedly downregulated the mRNA abundance of acetyl‐CoA carboxylase (ACC), lipoprotein lipase (LPL) and fatty acid‐binding protein 4 (FABP‐4) (p < .05), and upregulated the mRNA expression of hormone‐sensitive lipase (HSL), peroxisome proliferator‐activated receptor‐g coactivator‐1α (PGC‐1α), uncoupling protein 3 (UCP3) and liver carnitine palmitoyltransferase 1 (L‐CPT‐1) (p < .05). Therefore, our data suggest that protein‐restricted diets supplemented with optimal BCAA ratio, that is, 1 : 0.75 : 0.75–1 : 0.25 : 0.25, induce a shift from fatty acid synthesis to fatty acid oxidation in the liver of growing pigs. These effects may be associated with increased mitochondrial biogenesis.     

Evaluation of vegetable protein in canine diets: Assessment of performance and apparent ileal amino acid digestibility using a broiler model

Recent technological advances in the human food industry with respect to meat processing have decreased the availability of animal proteins to the pet food industry which typically formulates diets with an excess of animal protein. In the long term, this is not sustainable; thus, alternative protein sources need to be investigated. This study examined three canine diets, comparing a typical animal protein‐based diet (control) with two experimental diets where the animal protein was substituted in part with vegetable protein (formulated based either on total protein or amino acid content) using a broiler model. Each diet was fed to six cages each containing two birds from day 15, 18 cages in total (36 birds). Excreta were collected from days 19 to 21. On day 23, birds were euthanized and weighed, and their ileal digesta were collected and pooled for each cage. In addition, one leg per cage was collected for evaluation of muscle mass. Results showed no significant difference in animal performance (feed intake or live weight gain) or muscle to leg proportion across the diets. Birds fed the control diet and the diet balanced for amino acid content exhibited the greatest coefficients of apparent metabolizability for nitrogen (p < .001). Birds fed the diets that contained partial replacement of animal with vegetable protein generally had greater ileal digestibility of amino acids compared to birds fed the control (animal protein) diet. Analysis of excreta showed no dietary difference in terms of dry matter content; however, birds fed the diet balanced for total protein and the diet balanced for amino acid content had significantly greater excreta nitrogen than the control (p = .038). Overall, the study suggests vegetable proteins when formulated based on amino acid content are a viable alternative to animal proteins in canine diets.     

Dietary protein levels and amino acid supplementation patterns alter the composition and functions of colonic microbiota in pigs

Different dietary nitrogen (N) patterns may have different effects on gut microbiota. To investigate the effects of different crude protein (CP) levels or essential amino acids (EAA) supplementation patterns on the structure and functions of colonic microbiota, 42 barrows (25 ± 0.39 kg) were randomly assigned to 7 dietary treatments including: diet 1, a high CP diet with balanced 10 EAA; diet 2, a medium CP diet with approximately 2% decreased CP level from diet 1 and balanced 10 EAA; diets 3, 4, 5, 6 and 7, low CP diets with 4% decreased CP level from diet 1. Specifically, diet 3 was only balanced for Lys, Met, Thr and Trp; diets 4, 5 and 6 were further supplemented with Ile, Val and Ile + Val on the basis of diet 3, respectively; and diet 7 was balanced for 10 EAA. Results over a 110-d trial showed that reducing the CP level by 2% or 4% dramatically decreased N intake and excretion (P < 0.05) in the presence of balanced 10 EAA, which was not observed when altering the EAA supplementation patterns in low CP diet (−4%). With balanced 10 EAA, 2% reduction in dietary CP significantly reduced Firmicutes-to-Bacteroidetes (F:B) ratio and significantly elevated the abundance of Prevotellaceae NK3B31 (P < 0.05); whereas 4% reduction evidently increased the abundances of Proteobacteria, Succinivibrio and Lachnospiraceae XPB1014 (P < 0.05). Among the 5 low CP diets (−4%), supplementation with Ile, or Val + Ile, or balanced 10 EAA increased F:B ratio and the abundance of Proteobacteria. In addition, the predicted functions revealed that different CP levels and EAA balanced patterns dramatically altered the mRNA expression profiles of N-metabolizing genes, the “N and energy metabolism” pathways or the metabolism of some small substances, such as amino acids (AA) and vitamins. Our findings suggested that reducing the dietary CP levels by 2% to 4% with balancing 10 EAA, or only further supplementation with Ile or Val + Ile to a low protein diet (−4%) reduced the N contents entering the hindgut to various degrees, altered the abundances of N-metabolizing bacteria, and improved the abilities of N utilization.     

Effects of dietary protein level in the early fattening period on free amino acids and dipeptides in the blood and Longissimus thoracis muscle in Japanese Black steers

The effects of crude protein (CP) concentration in feed using soybean meal as its source in the early fattening period on the levels of free amino acids (FAAs) and dipeptides in the blood, and the levels in the M. longissimus thoracis after slaughter were studied in Japanese Black steers. Sixteen steers were divided into four groups and given feed with a CP content of 12, 14, 16, or 18% of dry matter (DM) from 10 to 20 months old, and they were fed with the same level of CP (13.5–13.9% of DM) until slaughter at 30 months of age. There was no significant difference in the weight gain, carcass weight or marbling score between the groups. Concerning the serum FAA and dipeptide contents at 20 months of age, the alanine, tyrosine and tryptophan levels decreased, while the carnosine (Car) level increased, with increases in the CP level in the feed. Although there were no significant differences in the FAA contents of the Longissimus thoracis muscle between the groups, the Car content decreased with increases in the feed protein level.     

CCK and GLP-1 release in response to proteinogenic amino acids using a small intestine ex vivo model in pigs

The impact of individual amino acids (AA) on gut hormone secretion and appetite regulation in pigs remains largely unknown. The aim of the present study was to determine the effect of the 20 proteinogenic AA on the release of the anorexigenic hormones cholecystokinin (CCK) and glucagon-like peptide 1 (GLP-1) in postweaning pigs. Six 25-d-old male piglets (Domestic Landrace × Large White; body weight = 6.94 ± 0.29 kg) were humanely killed for the collection of intestinal segments from the duodenum, jejunum, and ileum. Tissue samples from the three intestinal segments were used to determine which of the regions were more relevant for the analysis of gut peptides. Only the segments with the highest CCK and GLP-1 secretion and expression levels were evaluated with the 20 individual AA. Tissue segments were cut open, cleaned, and stripped of their muscle layer before identical circular samples were collected and incubated in 24-well plates for 1 h (37 °C, 5% v/v CO₂). The culture broth consisted of a glucose-free KRB buffer containing no added AA (control) or with the addition of 10 mM of 1 of the 20 proteinogenic AA. Following incubation, tissues and supernatant were collected for gene expression and secretion analysis of CCK and GLP-1 levels. CCK secretion and mRNA expression were higher (P < 0.05) in duodenum when compared with proximal jejunum or ileum, whereas GLP-1/proglucagon levels were higher in ileum vs. duodenum (P < 0.05) and jejunum (P < 0.05, for GLP-1 only) in postweaning pigs. Based on these results, the effect of AA on CCK and GLP-1 secretion was studied in the duodenum and ileum, respectively. None of the AA tested stimulated both anorexigenic hormones. Of all the essential AA, Ile, Leu, Met, and Trp significantly (P < 0.05) stimulated GLP-1 from the ileum, while only Phe stimulated CCK from the duodenum. Of the nonessential AA, amide AA (Gln and Asn) caused the release of CCK, while Glu and Arg increased the release of GLP-1 from the ileum. Interpreting the results in the context of the digestion and absorption dynamics, non-bound AA are quickly absorbed and have their effect on gut peptide secretion limited to the proximal small intestine (i.e., duodenum), thus, mainly CCK. In contrast, protein-bound AA would only stimulate CCK release from the duodenum through feedback mechanisms (such as through GLP-1 secreted mainly in the ileum).     

Corn protein has greater concentrations of digestible amino acids and energy than low-oil corn distillers dried grains with solubles when fed to pigs but does not affect the growth performance of weanling pigs

Three experiments were conducted to test the hypothesis that standardized ileal digestibility (SID) of amino acids (AA) and digestible energy (DE) and metabolizable energy (ME) in a new source of corn protein are greater than in corn distillers dried grains with solubles (DDGS) and that corn protein may be included in diets for weanling pigs. In experiment 1, the SID of AA was determined in two sources of DDGS (DDGS-1 and DDGS-2) and in corn protein. Results indicated that SID of most AA was greater (P < 0.05) in DDGS-2 and corn protein than in DDGS-1, but corn protein contained more digestible AA than both sources of DDGS. In experiment 2, the DE and ME in corn, the two sources of DDGS, and corn protein were determined. Results demonstrated that DE (dry matter basis) in corn protein was greater (P < 0.05) than in corn, but ME (dry matter basis) was not different between corn and corn protein. However, DE and ME in corn (dry matter basis) were greater (P < 0.05) than in DDGS-1 and DDGS-2. In experiment 3, 160 weanling pigs were allotted to four treatments in phases 1 and 2 and a common diet in phase 3. Corn protein was included at 5% to 10% in phases 1 and 2 at the expense of plasma protein and enzyme-treated soybean meal. Results indicated that although differences in average daily gain and gain to feed ratio were observed in phase 1, no differences among treatments were observed for the overall experimental period. In conclusion, the concentration of digestible AA is greater in corn protein than in DDGS; DE and ME in corn protein are also greater than in DDGS; and up to 10% corn protein may be included in phase 1 and phase 2 diets for weanling pigs.     

Influence of guanidination on apparent ileal digestibility of amino acids in pigs fed diets with soybean meal, rapeseed meal or peas as a protein source

The objective of this study was to determine the effect of guanidination, the conversion of dietary lysine to homoarginine, on the apparent ileal digestibilities (AID) of amino acids (AA) from 3 protein sources in diets for pigs. Six barrows, initial body weight of 18 ± 1.5 kg, fitted with a simple T-cannula at the distal ileum, were fed one of three corn starch-based diets that contained 18% crude protein (CP) from soybean meal, rapeseed meal or peas for an experimental period of 10 d according to a repeated 3 × 3 Latin square design. On d 9 of each experimental period, the pigs were fed one meal of the guanidinated test meals, which corresponded to their respective diets. In the test meals, 50% of the protein source in each diet was replaced, on an equal weight basis, with a guanidinated source. Guanidination increased the content of CP by 190, 116 and 28 g kg^− 1 for soybean meal, rapeseed meal and peas, respectively. With the exception of lysine, the AID of CP and AA were 2.2 to 8.2 percentage units (pu) higher in the test meal than in the soybean meal diet. Other than arginine and isoleucine, the differences were significant (P < 0.05) for all indispensable AA. With the exception of lysine, the AID of CP and AA were 3.8 to 11.9 pu higher in the test meal than in the diet with peas. The differences were significant (P < 0.05) for all indispensable AA. Other than lysine, glycine and proline, the AID of CP and AA were slightly higher, 0.1 to 4.3 pu, in the test meal than in the rapeseed meal diet. Of the indispensable AA, the differences were significant (P < 0.05) for arginine, leucine and phenylalanine. The AID of lysine was 10.9 pu lower (P < 0.05) in the test meal than in the rapeseed meal diet, 61.5 vs. 72.4%, indicating that guanidination of rapeseed meal was not uniform and random. The increase in the AID of CP and AA after guanidination may result from solubilisation of protein and degradation of antinutritional factors such as fiber, trypsin inhibitors and tannins.     

Ruminal degradability and intestinal digestibility of individual amino acids in mixed diets with different crude protein levels measured by the modified in vitro three‐step and mobile nylon bag technique

The ruminal degradability and intestinal digestibility of dry matter (DM), crude protein (CP) and amino acids (AA) in three total mixed rations with different CP levels were estimated using the modified in vitro three‐step procedure (TSP) and mobile nylon bag (MNB) technique on growing lambs. The ruminal effective degradability of DM and CP did not respond with increasing dietary CP level. However, the intestinal digestibility of DM was significantly increased with increasing dietary CP level estimated by TSP (P < 0.05) or MNB method (P < 0.01). Intestinal digestibility coefficients of CP determined by TSP were lower than those of the MNB method. Histidine was extensively degraded by rumen micro‐organisms, while tyrosine was the most anti‐degradable AA among the samples. The ruminal AA degradability exhibited no significant differences except for threonine, tryptophan, alanine, aspartic acid and proline for the three diets. Similarly, only a few AAs (i.e. histidine, methionine, tryptophan, aspartic acid and cysteine in TSP; histidine, tryptophan, aspartic acid and serine in MNB) had significant differences in their intestinal digestibility; in addition, values of MNB were lower than that of the TSP method, indicating that intestinal digestibility of DM seems to be overestimated in TSP, while that of CP might be overestimated in the MNB method.     

Intestinal acid flow, dry matter, starch and protein digestibility and amino acid absorption in beef cattle fed a high-concentrate diet with defluorinated rock phosphate, limestone or magnesium oxide

Five Angus heifers (285 kg) with duodenal and ileal cannulas were used to study the effects of feeding mineral acid neutralizers on digesta and fecal pH, intestinal acid flow, DM, starch and CP digestion and amino acid absorption in cattle fed a high-concentrate diet (7.5 kg/d). The experimental design was a 5 x 5 latin square. Treatments included control diet alone or control diet with the addition of 1.60% defluorinated rock phosphate-medium (DRP-M, 77% greater than 150 mu but less than 1,180 mu), 1.60% defluorinated rock phosphate-coarse (DRP-C, 85% greater than 850 mu but less than 1,700 mu), 1.28% limestone (92% greater than 150 mu but less than 850 mu) or .50% MgO, (81% greater than 250 mu but less than 1,180 mu), as an as-fed basis. Ileal pH was increased (P less than .05) from 7.49 to 7.85 by MgO. Fecal pH was increased (P less than .05) in this order: MgO (7.18) greater than DRP-M (6.86) = limestone (6.86) = DRP-C (6.75) greater than control (6.54). Liquid flow to the duodenum was increased (P less than .05) an average of 19.7% by the mineral treatments. Duodenal acid concentrations (mEq/liter) were greatest (P less than .05) for DRP-M (65.4) and DRP-C (64.0) vs limestone (55.5) and MgO (54.4). Acid flow to the intestine was increased (P less than .05) to 4.02, 4.53 and 4.82 Eq/d for animals fed limestone, DRP-M and DRP-C, respectively, from 3.51 Eq/d for control animals. Total tract DM, starch and CP digestibilities were similar (P greater than .10) among treatments. Limestone and DRP-M increased (P less than .10) the partial digestibility of most amino acids by about 6.8%. Mineral treatments did not increase total tract digestion of starch or CP. However, cattle fed DRP-M and limestone may have used protein more efficiently.     

Expression of selected genes encoding mechanistic pathways,nutrient and amino acid transporters in jejunum and ileum of broiler chickens fed a reduced protein diet supplemented with arginine,glutamine and glycine under stress stimulated by dexamethasone

Reducing crude protein and supplementation with synthetic amino acids in poultry nutrition is a recent trend to avoid wastage of protein and ammonia in production systems. Stress has been shown to impair intestinal barrier and increase inflammatory response. This study was performed on intestinal tissues of broiler chickens to understand the mechanism of stress induced by a synthetic glucocorticoid, dexamethasone (DEX) and the effect of supplementation of arginine, glutamine and glycine in reduced protein diets. Intestinal tissue samples from a previous study were utilized. Male Ross 308 chickens received a basal diet for the first seven days and then fed with crude protein that was reduced to 194 g/kg in grower experimental diets supplemented with glutamine, glycine and additional arginine at 10, 10 and 5 g/kg respectively. Half of the 96 individual birds were injected with DEX (0.5 mg/kg body weight) or saline on days 14, 16, 18 and 20 of age. mRNA expression for jejunum and ileum for amino acid transporters (y+LAT-1, B^o,+AT, EAAT-3 and CAT-1), mechanistic genes (SGLT-1, mTOR, IAP and FABP-2) and pro-inflammatory genes (MUC-2, NF-κB, iNOS, IL-8 and IL-1β) were analysed using real-time PCR. The results showed that DEX decreased y+ LAT1 in jejunum, B^o,+AT and EAAT-3 in ileum. Arginine increased CAT-1 in the jejunum and ileum under DEX treatment. Through an interaction, DEX reduced IAP in jejunum of glycine and arginine supplemented group and reduced mTOR in jejunum independently. DEX reduced MUC-2 and iNOS in jejunum and increased iNOS and IL8 in the ileum. Amino acid supplementation did not appear to ameliorate these effects; however, there were some positive effects of glycine on NF-κB and arginine through increased CAT-1. Mechanistic understanding of amino acid supplementation in broiler diets warrants further research particularly when dietary protein is reduced below the level tested in the present study.