Accelerated growth rate induced by neonatal high-protein milk formula is not supported by increased tissue protein synthesis in low-birth-weight piglets.

Low-birth-weight neonates are routinely fed a high-protein formula to promote catch-up growth and antibiotics are usually associated to prevent infection. Yet the effects of such practices on tissue protein metabolism are unknown. Baby pigs were fed from age 2 to 7 or 28 d with high protein formula with or without amoxicillin supplementation, in parallel with normal protein formula, to determine tissue protein metabolism modifications. Feeding high protein formula increased growth rate between 2 and 28 days of age when antibiotic was administered early in the first week of life. This could be explained by the occurrence of diarrhea when piglets were fed the high protein formula alone. Higher growth rate was associated with higher feed conversion and reduced protein synthesis rate in the small intestine, muscle and carcass, whereas proteolytic enzyme activities measured in these tissues were unchanged. In conclusion, accelerated growth rate caused by high protein formula and antibiotics was not supported by increased protein synthesis in muscle and carcass.

The protein level of isoenergetic formulae does not modulate postprandial insulin secretion in piglets and has no consequences on later glucose tolerance.

Early postnatal nutrition is involved in metabolic programming, an excess of protein being suspected to enhance early growth and the propensity to later develop insulin resistance and type 2 diabetes mellitus. The aim of the present study was to test the hypothesis that excessive protein intake during the suckling period would overstimulate the endocrine pancreas in the short term and alter durably its maturation, contributing to the later disruption of glucose homeostasis. Normal-birth-weight and low-birth-weight piglets were fed isoenergetic formulae providing an adequate-protein (AP, equivalent to sow milk) or a high-protein (HP, +48 %) supply between 7 and 28 d of age and were fed a standard diet until 70 d of age. During the formula-feeding period, the HP formula did not modify postprandial insulin secretion but transiently increased fasting insulin and the homeostasis model assessment-insulin resistance index (HOMA-IR, P < 0·05). Fasting insulin and HOMA-IR were restored to AP piglets' values 1 month after weaning. The structure of the endocrine pancreas was not affected by the protein content of the formula. The weight at birth had no major effect on the studied parameters. We concluded that a high-protein supply during the suckling period does not interfere with insulin secretion and endocrine pancreas maturation in the short term. It has no consequences either on glucose tolerance 1 month after weaning. The present study demonstrated that up-regulation of postprandial insulin secretion is not involved in higher growth observed in piglets fed a HP formula.

Growth, body composition and hormonal status of growing pigs exhibiting a normal or small weight at birth and exposed to a neonatal diet enriched in proteins.

Small birth weight and excess of early protein intake are suspected to enhance later obesity risk. The present study was undertaken to determine the impact of neonatal diets differing in protein content on growth, body composition and hormonal status of 70-d-old pigs born with normal weight (NW) or small weight (SW). At 7 d of age, male and female suckled piglets were assigned to the NW (approximately 1·4 kg at birth) or SW (approximately 0·99 kg at birth) groups. They were fed milk replacers formulated to provide an adequate protein (AP) or a high protein (HP) supply for 3 weeks. From weaning to 70 d of age, all animals received ad libitum the same standard diet. Growth rates were higher (P < 0·05) in HP piglets than in AP piglets during formula feeding and remained higher (P < 0·05) only in HP male pigs thereafter. No difference in feed consumption was detected between groups during the periods examined. Carcass lipid content and the relative weight of perirenal adipose tissue did not differ between the AP and HP pigs. Whereas plasma leptin concentration was higher (P < 0·05) in HP pigs than in AP pigs with a marked difference in SW pigs, plasma insulin-like growth factor (IGF)-I concentration and expression of IGF system genes were not affected by the diets. In summary, a HP intake during the suckling period induced an increase in growth rate that persisted only in male pigs during the post-weaning period. This response was not associated with any difference in adiposity parameters in this period.

Effect of milk formula protein content on intestinal barrier function in a porcine model of LBW neonates.

Our study aimed at investigating the impact of the level of protein in milk formula on intestinal structure, barrier function, and its nervous regulation in normal and LBW neonates using a porcine model. Normal birth weight (NBW) or LBW piglets were fed from d7 to d28 of age either with a high protein (HP) or with an adequate protein (AP) formula or stayed with their mother [mother fed (MF)]. The proximal jejunum and distal ileum were sampled at d28 for morphometry analysis and ex vivo permeability measurement in Ussing chambers. Formula feeding induced a trophic effect on the jejunum and ileum of both NBW and LBW piglets, which exhibited longer villi than MF animals, irrespective of the type of formula. In NBW piglets, intestinal permeability was not altered by formula feeding. On the contrary, LBW piglets fed with HP formula, but not AP, exhibited a greater ileal permeability than MF piglets. Feeding the HP formula also disturbed jejunal and ileal regulation of permeability by acetylcholine and vasoactive intestinal peptide (VIP) in LBW compared with MF LBW piglets. In conclusion, the level of protein in formulas did not modify intestinal structure and function in NBW individuals but dramatically modified intestinal barrier function physiology in LBW individuals.

A moderate threonine deficiency affects gene expression profile, paracellular permeability and glucose absorption capacity in the ileum of piglets.

High dietary threonine extraction by the digestive tract suggests that threonine contributes to maintain gut physiology. In the present study, we evaluated the impact of a low (6.5 g of threonine/kg diet; LT group) or a control well-balanced threonine diet (9.3 g of threonine/kg diet; C group) given to piglets for 2 weeks on ileal permeability and Na+-dependant glucose absorption capacity in Ussing chambers. The paracellular permeability was significantly increased in the ileum of LT compared to C piglets (P=.017). The Na+-dependent glucose absorption capacity showed a nonsignificant increase in the LT piglets. In addition, we analysed ileal gene expression profiles in the LT and C groups using porcine multitissue cDNA microarrays. Compared to the C piglets, the expression of 324 genes was significantly modified in the ileum of the LT piglets: 214 genes were overexpressed (145 annotated) and 110 were down-expressed (79 annotated). Among them, some are involved in immune and defense responses, energy metabolism and protein synthesis. Furthermore, microarray analysis highlights changes in the expression of the gene encoding for the sodium/glucose cotransporter (SGLT1) and of genes involved in the regulation of paracellular permeability (ZO-1, cingulin and myosin light chain kinase). In conclusion, our results indicate that a moderate threonine deficiency affects intestinal functionality.

Fatal effects of a neonatal high-protein diet in low-birth-weight piglets used as a model of intrauterine growth restriction.

BACKGROUND: Although full-term infants suffering intrauterine growth restriction (IUGR) are routinely fed high-protein (HP) formulas to ensure catch-up growth, the effects of HP intake are poorly understood. An IUGR piglet model provides an opportunity to investigate these effects. METHODS AND RESULTS: Twelve IUGR piglets were artificially fed HP formulas (50% more protein in comparison to sow milk) from the 2nd day of life (d2) until d28. Unexpectedly, all HP piglets developed poor growth, severe hypotonia and polypnea between d10 and d16. One third died spontaneously. This syndrome was investigated to understand its pathophysiology and to adopt a strategy to restore health. Blood and urine biochemistry and amino acid concentrations were investigated in 10 HP piglets and 8 piglets that were fed a normal-protein (NP) formula. In comparison to NP piglets, HP piglets showed significant hypokalemia (2.7 +/- 0.6 vs. 3.6 +/- 0.6 mmol/l; p < 0.01), hypophosphatemia (1.5 +/- 0.2 vs. 3.0 +/- 0.3 mmol/l; p > 0.01), hypercalcemia (3.0 +/- 0.3 vs. 2.5 +/- 0.2 mmol/l; p < 0.01), hyperammonemia (365 +/- 4 vs. 242 +/- 15 micromol/l; p < 0.05), elevated blood urea (6.5 +/- 0.4 vs. 1.3 +/- 0.4 mmol/l; p < 0.01) and elevated taurine concentrations (50.2 +/- 8.5 vs. 17.7 +/- 2.7 micromol/l; p < 0.01). CONCLUSIONS: These altered parameters indicated inadequate potassium and phosphorus dietary supplies in HP piglets. When the HP formula was supplemented with monocalcium phosphate and monopotassium phosphate (HP-sup), serum biochemistry was normalized in piglets fed this formula (n = 8). This experimental strategy restored growth in IUGR piglets fed HP-sup, without a toxic effect. The current findings suggest that use of an HP formula without a proportional increase in its phosphorus and potassium content induces pathology similar to the refeeding syndrome in IUGR piglets.

Comparative effect of orally administered sodium butyrate before or after weaning on growth and several indices of gastrointestinal biology of piglets.

Sodium butyrate (SB) provided orally favours body growth and maturation of the gastrointestinal tract (GIT) in milk-fed pigs. In weaned pigs, conflicting results have been obtained. Therefore, we hypothesised that the effects of SB (3 g/kg DM intake) depend on the period (before v. after weaning) of its oral administration. From the age of 5 d, thirty-two pigs, blocked in quadruplicates within litters, were assigned to one of four treatments: no SB (control), SB before (for 24 d), or after (for 11-12 d) weaning and SB before and after weaning (for 35-36 d). Growth performance, feed intake and various end-point indices of GIT anatomy and physiology were investigated at slaughter. The pigs supplemented with SB before weaning grew faster after weaning than the controls (P < 0.05). The feed intake was higher in pigs supplemented with SB before or after weaning (P < 0.05). SB provided before weaning improved post-weaning faecal digestibility (P < 0.05) while SB after weaning decreased ileal and faecal digestibilities (P < 0.05). Gastric digesta retention was higher when SB was provided before weaning (P < 0.05). Post-weaning administration of SB decreased the activity of three pancreatic enzymes and five intestinal enzymes (P < 0.05). IL-18 gene expression tended to be lower in the mid-jejunum in SB-supplemented pigs. The small-intestinal mucosa was thinner and jejunal villous height lower in all SB groups (P < 0.05). In conclusion, the pre-weaning SB supplementation was the most efficient to stimulate body growth and feed intake after weaning, by reducing gastric emptying and intestinal mucosa weight and by increasing feed digestibility.

Dietary and endogenous amino acids are the main contributors to microbial protein in the upper gut of normally nourished pigs.

Although amino acids (AA) synthesized by enteric microbiota in the upper gut of nonruminants can be absorbed, they do not necessarily make a net contribution to the host's AA supply. That depends on whether protein or nonprotein nitrogen sources are used for microbial protein production. We determined the contributions of urea, endogenous protein (EP), and dietary protein (DP) to microbial valine (M.VAL) at the distal ileum of growing pigs, based on isotope dilutions after a 4-d continuous infusion of l-[1-(13)C]valine to label EP and of [(15)N(15)N]urea. Eight barrows were assigned to either a cornstarch and soybean meal-based diet with or without 12% added fermentable fiber from pectin. Dietary pectin did not affect (P > 0.10) the contributions of the endogenous and DP to M.VAL. More than 92% of valine in microbial protein in the upper gut was derived from preformed AA from endogenous and DP, suggesting that de novo synthesis makes only a small contribution to microbial AA.

Intestinal physiology and peptidase activity in male pigs are modulated by consumption of corn culture extracts containing fumonisins.

Fumonisin B(1) (FB1) alters intestinal epithelial cell cycle and absorptive, secretory, and barrier properties in vitro, but in vivo data are lacking. Therefore, we tested the hypothesis that repeated intake of a corn culture extract rich in fumonisins, mainly in FB1, alters indices of intestinal absorptive and secretory physiology and barrier function in vivo. Intra-litter pairs of pigs (n = 36) weaned at 28 d, were fed the vehicle (control) or the extract (providing 1.5 mg FB1/kg body weight) daily for 9 d starting 7 d postweaning. After slaughter, the jejunal mucosa of pigs was mounted in Ussing chambers (UC). Extract consumption for 9 d decreased the gain:feed ratio (P = 0.04) and increased liver weight (P = 0.01). Basal net ion secretion (P = 0.02), sodium-dependent glucose absorption (P = 0.02), and theophylline-induced secretion (P < 0.01) of the jejunal mucosa determined in UC were higher in pigs fed the extract than in controls. By contrast, jejunal permeability to the horseradish peroxidase model protein in UC was not influenced by extract consumption. Ileal villi tended to be longer (P = 0.07) and jejunal aminopeptidase N activity was lower (P < 0.01) in pigs fed the extract. In conclusion, consumption of an extract rich in fumonisins for 9 d has the potential to alter intestinal physiology, villous architecture, and enzyme activities. Underlying mechanisms remain to be investigated.

Impact of intrauterine growth retardation and early protein intake on growth, adipose tissue, and the insulin-like growth factor system in piglets.

Small birth weight and excess of early protein intake are suspected to enhance later adiposity. The present study was undertaken to determine the impact of diets differing in protein content on short-term growth, adipose tissue development, and the insulin-like growth factor (IGF) system in piglets. Normal (NW) and small (SW) birth weight piglets were fed milk-replacers formulated to provide an adequate (AP) or a high protein (HP) supply between 7 and 28 d of age. The fractional growth rate was higher (p < 0.01) in SW than in NW piglets. At 7 d of age, the lower (p < 0.05) weight of perirenal adipose tissue relative to body mass in SW than in NW piglets did not involve significant changes in plasma IGF-I, leptin, or insulin-like growth factor binding protein levels, but involved differences (p < 0.05) in the expression of IGF-I and leptin in adipose tissue. Growth rates did not differ between AP and HP piglets. At 28 d of age, HP piglets had lower (p < 0.001) relative perirenal adipose tissue weight but did not differ clearly from AP piglets with regard to the IGF system. It remains to be determined whether piglets fed such a high protein intake will stay subsequently with a low adiposity.

A moderate threonine deficiency differently affects protein metabolism in tissues of early-weaned piglets.

A moderate threonine deficiency may affect differently tissue protein metabolism. In this study, we compared protein metabolism in the small and large intestines, the liver, and the carcass of piglets (Sus scrofa) pair-fed either a control well-balanced diet (C: 9.3 g threonine/kg diet) or a low threonine diet (LT: 6.5 g threonine/kg diet) for 2 weeks. In the small intestine, the LT diet did not modify protein deposition, fractional protein synthesis rate (K(S)) and AA protein composition. Ubiquitin mRNA level, a component of the ubiquitin-dependent proteolytic pathway, was significantly decreased in the jejunum of the LT piglets. Protein deposition measured in the carcass and the colon, and K(S) measured in the semitendinosus muscle and the colon, did not differ between LT and C piglets. Nevertheless, in these compartments, threonine content was reduced indicating deposition of proteins less rich in threonine. In the liver, protein retention was reduced, K(S) was increased and AA protein composition was modified in the LT compared to the C piglets. In conclusion compared to the other compartments, small intestinal protein metabolism seems to be less sensitive to a moderate dietary threonine deficiency. This indicates that dietary threonine extraction by the small intestine may reduce threonine availability for the other tissues when young piglets were fed a diet marginally deficient in threonine.

Tryptophan metabolism and related B vitamins in the multiparous sow fed ad libitum after farrowing.

Although dietary content of tryptophan has been related to variations of feed intake in lactating sows, the mechanisms remain to be elucidated. Twenty multiparous crossbred Landrace × Large White sows were used to assess variations of tryptophan metabolism around farrowing. Sows were fed 3 kg/d of a standard gestation diet from insemination until farrowing. They were then fed ad libitum a standard lactation diet until weaning. Sows were catheterised on day 70 of gestation and blood samples were drawn on day 37 before parturition, daily during the week before and the week after farrowing, and on days 14 and 21 of lactation. Plasma concentrations of amino acids, kynurenine, niacin, haptoglobin, urea, and vitamin B6 concentration in red blood cells were determined. During the week following parturition, plasma tryptophan and niacin decreased while plasma kynurenine increased (p < 0.05). On the 2nd and 3rd weeks of lactation, plasma tryptophan and kynurenine returned to pre-farrowing concentrations, while niacin increased throughout lactation (p < 0.05). Vitamin B6 increased progressively during the week after farrowing (p < 0.05) and remained constant at a high concentration thereafter. The average feed intake of the sow during lactation was positively correlated with the mean concentrations of niacin (r(2) = 0.25; p < 0.001) and kynurenine (r(2) = 0.31; p < 0.001) in plasma and with vitamin B6 in red blood cells (r(2) = 0.68; p < 0.001). This study suggests that tryptophan catabolism presumably through the kynurenine pathway is high during the 1st week after farrowing, and that dietary supply of niacin and vitamin B6 could be transiently suboptimal in early lactation.

Main intestinal markers associated with the changes in gut architecture and function in piglets after weaning.

We analysed the spatio-temporal sequence of events concerning the morphology, physiology and ecology of the gut of piglets during the 2 weeks following weaning, in order to provide a limited number of variables that could be relevant markers of the gut post-weaning changes. An experiment was conducted on sixty piglets fasted for 2 d, then administered a weaning diet with a moderate or a high content of wheat using controlled gastric feeding, and slaughtered at different time-points post-weaning. Sixty-nine variables were analysed by principal component analysis. The results showed that the temporal changes induced in the gut by weaning can be divided into two periods: an acute period happening immediately after weaning, followed after day 5 by a more progressive adaptative and maturational phase. The main factors of this adaptation were the refeeding process and the time, while the diet per se had little influence. The villus length, lactose activity, macromolecule fluxes across the jejunum and the plasma cholecystokinin were proposed as markers of the acute phase. Ths mass of the jejunum, the weight of the pancrease, the content of stomach, the trypsin activity and the theophyl-line-induced secretion in jejunum were related to the re-feeding. Markers proposed to follow the gut maturation were the maltase activity, the glucose absorption and the basal resistance in the ileum, the lactobacilli and enterococci in the colon, and the pH of colonic and caecal contents. These markers might be helpful to design suitable diets to limit posts-weaning gut disorders in pigs.

Effects of a bovine colostrum-supplemented diet on some gut parameters in weaned piglets.

The present study investigated the effects of a bovine colostrum-supplemented diet on gut post-weaning adaptation and health in piglets. Thirty-six 21-d-old piglets were allocated to one of the three following dietary treatments: sow-reared (SR), weaned on a control starter diet (WCtrl) or on a starter diet supplemented with bovine colostrum (WCol) until slaughter at 28 d or 35 d of age. Gastric pH and intestinal bacteriological, structural and functional parameters were determined. Compared to WCtrl, the gastric pH was lower (P < 0.05) and the duodenal lactobacilli:coliform ratio was higher (P = 0.05) in WCol piglets. The relative small intestine weight was 18% (P < 0.05) higher in WCol piglets than in SR piglets. Duodenal villous height was lower (P < 0.01) in WCtrl than in SR piglets, whereas the value for WCol piglets was intermediate. The weaning-increased crypt cell proliferation was not affected by bovine colostrum supplementation. The mucosal ribosomal capacity was higher (P < 0.05) in W than in SR piglets. In conclusion, a diet supplemented with colostrum induced, although not always significantly, variations of gut parameters, suggesting that globally, colostrum may limit weaning-induced gut structural and microbial alterations. The observed effects occurred early and were maintained throughout the post-weaning adaptive phase.

High-viscosity carboxymethylcellulose reduces carbachol-stimulated intestinal chloride secretion in weaned piglets fed a diet based on skimmed milk powder and maltodextrin.

High-viscosity carboxymethylcellulose (CMC) promotes gastrointestinal disorders, tissue alterations and bacterial overgrowth in pigs. The impact of CMC on intestinal absorptive and secretory physiology is not known. We hypothesised that CMC consumption alters intestinal Na-dependent glucose absorption and stimulates electrogenic chloride secretion. For testing this hypothesis, twenty-four piglets were weaned at 21 d of age and pair-fed for 13 d a starter diet based on skimmed milk powder and maltodextrin containing cellulose (control) or CMC. Body weight and faecal total aerobe and coliform counts were measured kinetically. At slaughter, digesta were weighed and characterised for viscosity and pH. Gastrointestinal tissues were weighed and sampled for physiology in Ussing chambers, morphometry and enzymology. Glucose absorption tended to be higher (P = 0.08) and carbachol-stimulated chloride secretion was lower (P = 0.01) with CMC in the small intestine, without changes in the colon. Aerobes were transiently higher at day 7 (P < 0.05) but coliform counts remained unchanged (P = 0.78) and beta-haemolitic Escherichia coli were virtually absent. Stomach and small-intestinal segments were heavier, and viscosity higher with CMC (0.001 < P < 0.05). The pH in the stomach was higher, and in the caecum and proximal colon lower with CMC (0.001 < P < 0.05). Jejunal villus area was slightly reduced with CMC (P < 0.05) without effects on enzyme activities (P > 0.10). In conclusion, CMC supplementation had pro-absorptive effects on the small intestine, possibly due to the absence of pathogenic E. coli in the present study.

Importance of sanitary environment for growth performance and plasma nutrient homeostasis during the post-weaning period in piglets.

Deterioration of sanitary conditions in piggeries is known to limit growth performance through inducing a moderate immune response. This article reports the results of an experiment performed to reproduce the consequences of bad sanitary conditions on growth performance and nutrient plasma concentrations of piglets after weaning. We propose to use these experimental conditions as a model for studying the interactions between nutrition and pig health. In this experiment, 20 pairs of littermate piglets were selected and weaned at 28 days of age on the basis of their body weight. Within each pair, piglets were pair-fed and each one was affected to one of the two experimental groups. The first group was housed in a clean environment and was fed an antibiotic supplemented standard diet. The second group was kept in unsanitary rooms, mixed with non-experimental piglets and was fed the same standard diet but without antibiotic supplementation. Compared to pigs kept in the clean environment, piglets kept in the unsanitary environment had significantly lower rate of weight gain and feed efficiency from weaning to 20 d post weaning then from 36 - 45 d post weaning. They also displayed higher plasma concentrations of haptoglobin, copper, vitamin B12 and lysine but lower concentrations of glutathione, pyridoxal-5-phosphate, folic acid, threonine and tryptophan. Our results showed that a reduction of growth performance and a modification of nutrient utilization can be induced by decreasing the sanitary quality of environment where pigs are kept after weaning and after transition to another building. This response could be explained by a moderated activation of body defences.

Is tryptophan catabolism increased under indoleamine 2,3 dioxygenase activity during chronic lung inflammation in pigs?

In a preliminary study we observed that piglets suffering from chronic lung inflammation induced by an intravenous injection of complete Freund adjuvant showed a marked decrease in plasma tryptophan (Trp) concentration suggesting increased Trp utilisation. During the inflammatory process, a cytokine-induced enzyme called indoleamine 2,3-dioxygenase (IDO) has been shown to catabolise Trp into kynurenine (Kyn). Yet, during inflammation, increased Trp catabolism may decrease Trp availability for other functions such as growth. This metabolic pathway has never been studied in pigs. So, the objectives of this study were to measure IDO activity in pigs and to determine if the decrease in plasma Trp concentrations previously observed in piglets suffering from chronic lung inflammation could be explained by the induction of IDO activity. In order to do so, we compared IDO activity measured in the tracheo-bronchial lymph nodes and in the lungs of 7 piglets, injected with complete Freund adjuvant (CFA), to 7 pair-fed littermate healthy controls. Blood samples were taken at 0, 2, 5, 7 and 10 days following CFA injection in order to measure plasma Trp, Kyn and haptoglobin concentrations. Indoleamine 2,3-dioygenase activity in the tracheo-bronchial lymph nodes (P < 0.05), in the lungs (P < 0.07) and plasma haptoglobin (P < 0.01) were higher in pigs with lung inflammation than in the controls. Plasma Trp and Kyn were not significantly affected by CFA injection. Our data showed that IDO is activated under chronic lung inflammation in pigs. The impact of IDO activation on plasma Trp concentration and its availability is discussed according to the amount of Trp provided by the diet.

Catabolism through the threonine dehydrogenase pathway does not account for the high first-pass extraction rate of dietary threonine by the portal drained viscera in pigs.

In pigs the extensive threonine utilization by the splanchnic tissues explains the relative inefficiency of dietary threonine conversion for body protein accretion. Two experiments were conducted to estimate the contribution of the portal drained viscera (PDV) and the liver to threonine metabolism and especially catabolism in growing pigs. In the first experiment, four pigs were surgically prepared for chronic catheter insertion in the portal, hepatic and jugular veins and in the carotid artery. They were continuously infused with L-[1-(13)C]threonine through the jugular catheter. The PDV and total splanchnic viscera (PDV and the liver) extracted 14.3 and 18.8% of arterial threonine input, respectively. In a second experiment, we studied the metabolism of dietary threonine in the PDV and the liver in six female growing pigs. Animals were surgically prepared as in the first experiment except that L-[1-(13)C]threonine and [(15)N]glycine were continuously infused in the duodenum for 10 h. Unlabelled and labelled threonine and glycine PDV, liver and splanchnic tissues balance were calculated from plasma samples taken during the last 2 h of this infusion. Splanchnic tissues extracted 60% of infused labelled threonine, 88% of which was extracted by PDV so that threonine extraction by the liver was low. Both the liver and the pancreas can degrade threonine through the L-threonine 3-dehydrogenase pathway but not the intestine. Our data suggest that threonine catabolism through the L-threonine 3-dehydrogenase pathway was only a minor component of total threonine utilization in the splanchnic tissues.

Identification of dietary and endogenous ileal protein losses in pigs by immunoblotting and mass spectrometry.

Ileal flows and the endogenous or dietary origin of soluble proteins present in ileal digesta were determined in pigs fed diets containing different pea cultivars (Solara, Madria and Eiffel) and micro-ground peas (c.v. Solara). Ileal digesta proteins were analyzed by electrophoresis and densitometry analysis and were identified by LC-MS-MS spectrometry and immunoblotting. The ileal flows of proteins differed (P < 0.1) among the 3 pea cultivars; the flow in pigs fed the Madria-containing diet was higher than that of pigs fed the Eiffel- and Solara-containing diets. The flow was reduced by micro-grinding the peas. The true digestibility of pea proteins and the endogenous losses were not correlated. However, at this intestinal level, protein losses were essentially of endogenous origin (enzymes, antibodies), and from the partly digested pea albumin fraction. Pea lectin and albumin PA1b were totally resistant to gastric and small intestinal digestion and a minor resistant peptide from pea albumin PA2 was detectable. In contrast, the storage proteins, legumin and vicilin, were not detectable by antibodies or by LC-MS-MS.

Effects of grinding and thermal treatments on hydrolysis susceptibility of pea proteins (Pisum sativum L.).

The effects of three particle sizes with two types of grindings and two thermal treatments on pea protein extraction (PE) and susceptibility to in vitro enzymatic hydrolysis (pepsin plus trypsin) were studied. Degrees of hydrolysis (DH) were calculated. Remaining peptides were detected by SDS-PAGE and identified by immunoblotting and MS/MS spectrometry. The increase in particle size decreased PE and DH due to a restricted access of solvents and enzymes to proteins. The thermal treatment induced a decrease in PE but did not modify DH. Heating improved legumin (alphaM) and convicilin pepsin hydrolyses but reduced the pea albumin 2 (PA2) hydrolysis. After pepsin and trypsin hydrolysis, only peptides from vicilin and lectin were identified by LC-MS/MS analyses, whatever the treatment.