To explore the effect of manno-oligosaccharide (MOS) on intestinal health in weaned pigs upon enterotoxigenic Escherichia coli K88 (ETEC) challenge, thirty-two male weaned pigs were randomly assigned into four groups. Pigs fed with a basal diet or basal diet containing MOS (0·6 g/kg) were orally infused with ETEC or culture medium. Results showed that MOS significantly elevated the digestibility of crude protein and gross energy in both ETEC-challenged and non-challenged pigs (P < 0·05). MOS also elevated serum concentrations of IgA and IgM (P < 0·05), but decreased serum concentrations of TNF-α, IL-1β and IL-6 (P < 0·05) in ETEC-challenged pigs. Interestingly, MOS increased villus height and the ratio of villus height:crypt depth in duodenum and ileum (P < 0·05). MOS also increased duodenal sucrase and ileal lactase activity in ETEC-challenged pigs (P < 0·05). MOS decreased the abundance of E. coli, but increased the abundance of Lactobacillus, Bifidobacterium and Bacillus in caecum (P < 0·05). Importantly, MOS not only elevated the expression levels of zonula occludens-1 (ZO-1), claudin-1 and GLUT-2 in duodenum (P < 0·05) but also elevated the expression levels of ZO-1, GLUT-2 and L-type amino acid transporter-1 in ileum (P < 0·05) upon ETEC challenge. These results suggested that MOS can alleviate inflammation and intestinal injury in weaned pigs upon ETEC challenge, which was associated with suppressed secretion of inflammatory cytokines and elevated serum Ig, as well as improved intestinal epithelium functions and microbiota.

Glutathione (GSH) is considered to play an important role in maintaining the integrity of the small intestine. In piglets, altered mucosal GSH levels might therefore be involved in weaning-induced changes of the small intestinal morphology and barrier function. To test this hypothesis, we aimed to challenge the mucosal GSH redox status during the first 28 days after weaning, by feeding diets containing 5% fresh linseed oil (CON), or 2.5% (OF1) or 5% (OF2) peroxidized linseed oil (peroxide value 225 mEq O2/kg oil) and exploring the effects on gut integrity. Piglets were pair-fed and had a total daily feed allowance of 32 g/kg BW. A fourth treatment included animals that were fed the control diet ad libitum (ACON). Animals were sampled at days 5 and 28 post-weaning. The malondialdehyde (MDA) concentration and GSH redox status (GSH/GSSG Eh) were determined in blood, liver and small intestinal mucosa. Histomorphology of the duodenal and jejunal mucosa was determined, and Ussing chambers were used to assess fluorescein isothiocyanate dextran (FD4) and horseradish peroxidase (HRP) fluxes across the mucosa. Results show that peroxidized linseed oil imposed an oxidative challenge at day 28, but not at day 5 post-weaning. At day 28, increasing levels of dietary peroxides to pair-fed pigs linearly increased MDA levels in duodenal and jejunal mucosa. Moreover, FD4 fluxes were significantly increased in OF1 (+75%) and OF2 (+64%) in the duodenum, and HRP fluxes tended (P=0.099) to show similar differences, as compared to CON. This co-occurred with a significant 11 mV increase of the hepatic GSH/GSSG Eh, potentiated by a significantly increased GSH peroxidase activity for treatments OF1 (+47%) and OF2 (+63%) in liver as compared to CON. Furthermore; duodenal HRP flux significantly correlated with the hepatic glutathione disulphide (GSSG) level (r=0.650), as also observed in the jejunum for hepatic GSSG (r=0.627) and GSH/GSSG Eh (r=0.547). The jejunal permeability was not affected, but FD4 and HRP fluxes significantly correlated with the local GSH (r=0.619; r=0.733) and GSSG (r=0.635; r=0586) levels. Small intestinal histomorphology was not affected by dietary lipid peroxides, nor were there any correlations found with the GSH redox system. To conclude, under oxidative stress conditions, jejunal barrier function is related to the local and hepatic GSH redox system. It is suggested that the hepatic GSH system participates in the elimination of luminal peroxides, and thereby impacts on duodenal barrier function.

An experimental, microbially enhanced soybean product (MEPRO) was evaluated as a replacement for fishmeal (FM). Assessment of feedstuffs should include estimation of digestibility as well as pig performance and in combination with dietary additives. Digestibility values determined in growing pigs may not apply to nursery pigs; thus, standardized ileal digestibility (SID) of amino acids (AA) in MEPRO and FM were determined using 30±1.6 kg BW ileal-cannulated barrows (n=6) and 9.8±1.2 kg BW barrows (n=37; serial slaughter). Experimental diets included MEPRO, FM and nitrogen free where FM and MEPRO were included as the sole protein source. The SID of AAs was 3% to 5% lower in MEPRO than FM when fed to 30 kg pigs. The SID of arginine and methionine was greater (P<0.05) in MEPRO than FM when fed to 10 kg pigs. The SID of AAs was 12% to 20% lower in FM when fed to 10 v. 30 kg pigs but only 3% to 9% lower in MEPRO. A total of 336 barrows and gilts were weaned at 21 days of age (initial BW=6.1±0.8 kg) and used in a performance trial. Pens of pigs were assigned to one of the six experimental diets (8 pens/diet in two blocks). Treatment diets were fed in Phase I (7 days) and Phase II (14 days) with all pigs fed a common Phase III diet (14 days). Experimental diets included (1) negative control (NEG) containing corn, soybean meal and whey, (2) NEG+acidifier, (3) NEG+FM (POS), (4) POS+acidifier (POS A+), (5) NEG+MEPRO (MEPRO) and (6) MEPRO+acidifier. The FM and MEPRO were included at 7.5% and 5.0% in Phase I and II diets, respectively. Diets were formulated to meet the standard nutrient requirements for weaned pigs. Pig BW and feed disappearance was measured weekly and fecal scores were measured daily for the first 14 days post-weaning as an indicator of post-weaning diarrhea syndrome (PWDS). Performance (BW, daily gain, feed intake and gain : feed) was not significantly different among treatments. Treatment for PWDS occurred on different days in each block. Analysis of fecal score was completed separately by block. Pigs fed the NEG diets had higher (P=0.02) fecal scores than pigs fed the POS diets on days 2 and 3 (block 1) and higher (P<0.05) than pigs fed MEPRO or POS diets and diets with dietary acidifier on days 6 and 3 (block 2). The MEPRO holds promise as an alternative to FM in nursery pig diets.

Stress induces injury in intestinal barrier function in piglets. Long-chain n-3 PUFA have been shown to exhibit potential immunomodulatory and barrier protective effects in animal models and clinical trials. In addition, corticotropin-releasing hormone (CRH)/CRH receptor (CRHR) signalling pathways play an important role in stress-induced alterations of intestinal barrier function. We hypothesised that fish oil could affect intestinal barrier function and CRH/CRHR signalling pathways. In total, thirty-two weaned pigs were allocated to one of four treatments. The experiment consisted of a 2×2 factorial design, and the main factors included immunological challenge (saline or lipopolysaccharide (LPS)) and diet (5 % maize oil or 5 % fish oil). On d 19 of the trial, piglets were treated with saline or LPS. At 4 h after injection, all pigs were killed, and the mesenteric lymph nodes (MLN), liver, spleen and intestinal samples were collected. Fish oil decreased bacterial translocation incidence and the number of translocated micro-organisms in the MLN. Fish oil increased intestinal claudin-1 protein relative concentration and villus height, as well as improved the intestinal morphology. In addition, fish oil supplementation increased intestinal intraepithelial lymphocyte number and prevented elevations in intestinal mast cell and neutrophil numbers induced by LPS challenge. Moreover, fish oil tended to decrease the mRNA expression of intestinal CRHR1, CRH and glucocorticoid receptors. These results suggest that fish oil supplementation improves intestinal barrier function and inhibits CRH/CRHR1 signalling pathway and mast cell tissue density.

Two experiments were conducted to investigate if adjustments in diet formulations either based on total analysed amino acids or standardized ileal digestible (SID) amino acids may be used to eliminate negative effects of including heat-damaged soybean meal (SBM) or heat-damaged corn distillers dried grains with solubles (DDGS) in diets fed to weanling pigs. In Experiment 1, four corn–SBM diets were formulated. Diet 1 contained non-autoclaved SBM (315 g/kg), and this diet was formulated on the basis of analysed amino acid concentrations and using SID values from the AminoDat® 4.0 database. Diet 2 was similar to Diet 1 in terms of ingredient composition, except that the non-autoclaved SBM was replaced by autoclaved SBM at 1 : 1 (weight basis). Diet 3 was formulated using autoclaved SBM and amino acid inclusions in the diet were adjusted on the basis of analysed total amino acid concentrations in the autoclaved SBM and published SID values for non-autoclaved SBM (AminoDat® 4.0). Diet 4 also contained autoclaved SBM, but the formulation of this diet was adjusted on the basis of analysed amino acids in the autoclaved SBM and SID values that were adjusted according to the degree of heat damage in this source of SBM. Pigs (160; initial BW: 10.4 kg) were allotted to the four treatments with eight replicate pens per treatment in a randomized complete block design. Diets were fed to pigs for 21 days. The gain to feed ratio (G : F) was greater (P<0.05) for pigs fed Diet 1 compared with pigs fed the other diets and pigs fed Diet 4 had greater (P<0.05) G : F than pigs fed Diet 2. In Experiment 2, 144 pigs (initial BW: 9.9 kg) were allotted to four diets with eight replicate pens per diet. The four diets contained corn, SBM (85 g/kg) and DDGS (220 g/kg), and were formulated using the concepts described for Experiment 1, except that heat-damaged DDGS, but not heat-damaged SBM, was used in the diets. Pigs fed Diet 1 had greater (P<0.05) G : F than pigs fed Diet 2, but no differences were observed for G : F among pigs fed diets containing autoclaved DDGS. Results demonstrate that the negative effects of heat damage of SBM or DDGS may be ameliorated if the reduced concentration and digestibility of amino acids in heat-damaged SBM or DDGS is taken into account in diet formulation. Further research is needed to improve the prediction of the ileal digestibility of amino acids in heat-processed ingredients used in practical diet formulations.

The present study was conducted to investigate the effects of chitosan (CS)-Zn on intestinal morphology, mucosal epithelial cell apoptosis and mucosal immune function in weanling pigs. A total of 150 weanling barrows with a body weight of 7·2 kg were randomly allocated into five groups. A basal diet without Zn supplementation was used as the control and other four groups were fed the control diet supplemented with 50 or 100 mg/kg of Zn as CS-Zn, 100 mg/kg of Zn as ZnSO4 and 3000 mg/kg of Zn as ZnO, respectively. The feeding trial lasted for 28 d. The results showed that serum diamine oxidase activities, d-lactate levels and endotoxin contents were lower in pigs fed dietary 100 mg/kg of Zn as CS-Zn or 3000 mg/kg of Zn as ZnO than in pigs fed the control or 100 mg Zn/kg as ZnSO4 diet. The ratios of the villus height:crypt depth of the duodenum, jejunum and ileum were higher in pigs that received 100 mg/kg of Zn as CS-Zn or a high level of Zn as ZnO than in pigs fed the control diet. Moreover, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL)-stained ileal epithelial cells were found in the control group, and apoptotic cells did not appear prominently in pigs that received the 100 mg/kg of CS-Zn or ZnO diet. Secretory IgA concentration in ileal mucus was increased in the dietary group that received 100 mg/kg of CS-Zn or ZnO. These results indicated that dietary 100 mg CS-Zn/kg had similar biological effects to dietary 3000 mg ZnO/kg on intestinal morphology, mucosal epithelial cell apoptosis and mucosal immune function.

A 2 × 2 factorial experiment was conducted to investigate the interactions between laminarin (LAM; 0 and 300 parts per million (ppm)) and fucoidan (FUC; 0 and 240 ppm) levels on intestinal morphology, selected microbiota and inflammatory cytokine gene expression in the weaned pig. There was an interaction between LAM and FUC supplementation on the Enterobacteriaceae population (P< 0·05) and the abundance of attaching and effacing Escherichia coli (AEEC) strains (P< 0·05) in the colon. Pigs offered the FUC diet had a reduced Enterobacteriaceae population compared with pigs offered the basal diet. However, the effect of FUC on the Enterobacteriaceae population was not observed when combined with LAM. Pigs offered the LAM diet had reduced abundance of AEEC strains compared with pigs offered the basal diet. However, there was no effect of LAM on the abundance of AEEC strains when combined with FUC. There was an interaction between LAM and FUC supplementation on villous height (P< 0·01) and the villous height:crypt depth ratio (P< 0·01) in the duodenum. Pigs offered the LAM or FUC diet had an increased villous height and villous height:crypt depth ratio compared with pigs offered the basal diet. However, there was no effect of the LAM and FUC combination diet on intestinal morphology. Pigs offered the LAM-supplemented diets had a lower IL-6 (P< 0·05), IL-17A (P< 0·01) and IL-1β (P< 0·01) mRNA expression in the colon compared with pigs offered the diets without LAM. In conclusion, supplementation with either LAM or FUC alone modified intestinal morphology and selected intestinal microbiota, but these effects were lost when offered in combination.

The study evaluated whether feeding diosmectite–ZnO composite (DS-ZnO) at 500 mg Zn/kg to early weaned pigs would alleviate the weaning-related intestinal disorders as a substitute for high concentration of ZnO (2250 mg Zn/kg). The pigs weaned at an age of 21 ± 1 d were allotted to four treatments groups as follows: (1) control; (2) DS-ZnO, 500 mg Zn/kg diet; (3) ZnO, 2250 mg Zn/kg diet; and (4) mixture of 2·0 g DS/kg and 500 mg Zn/kg from ZnO (equal amount of DS and ZnO in the DS-ZnO treatment group). The results showed that, compared with the control on days 7 and 14 post-weaning, addition of DS-ZnO at 500 mg Zn/kg improved (P< 0·05) daily gain and feed intake, decreased (P< 0·05) post-weaning scour scores, increased (P< 0·05) jejunal villus height and the ratio of villus height and crypt depth, decreased (P< 0·05) jejunal paracellular permeability of fluorescein isothiocyanate dextran 4 kDa and up-regulated (P< 0·05) tight junction protein expression of occludin, claudin-1 and zonula occludens-1 in jejunal mucosa. The mRNA levels of TNF-α, IL-6 and interferon-γ (IFN-γ) on day 7 post-weaning were also decreased (P< 0·05). The piglets fed DS-ZnO at 500 mg Zn/kg did not differ in the above parameters from those fed ZnO at 2250 mg Zn/kg, while they had better performance than those fed the mixture of DS and ZnO. Supplementation with DS-ZnO at 500 mg Zn/kg was effective in alleviating diarrhoea, barrier dysfunction and inflammatory cytokine expression and up-regulating tight junction protein expression.

Starch is the major energy source for monogastric mammals and humans. The present study was conducted to evaluate the liver metabolic responses of weaned pigs fed with different dietary starches. A total of sixteen weaned pigs were fed with two experimental diets containing either cassava starch (CS, 80 % amylopectin and 20 % amylose) or maize starch (70 % amylopectin and 30 % amylose). The present results showed that the growth performance was not affected by different dietary starches (P>0·05). However, ingestion of CS not only increased the lipid content in liver tissues, but also elevated the concentrations of serum cholesterol and insulin (P < 0·05). The metabolic responses induced by CS were associated with more lipogenic enzymes such as fatty acid synthase and 3-hydroxy-3-methyl-glutaryl-CoA reductase in liver (P < 0·05). Real-time PCR quantification for lipid metabolic genes indicated that ingestion of CS not only up-regulated the expression of these lipogenic genes, but also decreased the expression of lipolytic genes. These results suggested that the metabolic responses of weaned pigs fed with different dietary starches may vary widely depending on their composition, and ingestion of starches that are high in amylopectin may produce a stronger insulinaemic response and lead to an up-regulation of lipogenesis in the liver.

The present study was conducted to investigate the effect of digestion rate of dietary starch on postprandial systemic circulating glucose, insulin and lipid profiles, and the activity and gene expression of lipid metabolism-related enzymes in weaned pigs. A total of twenty-four weaned pigs, surgically fitted with a catheter in the jugular vein, were randomly assigned to three dietary treatment groups, representing the high digestion rate starch (HDRS) group, the moderate-digestion rate starch (MDRS) group and the low-digestion rate starch (LDRS) group. The amylopectin:amylose ratios in the diets of each group were 27·6:1, 27·6:8·5 and 1:27·6, respectively. The serum concentrations of glucose, TAG, total cholesterol, LDL-cholesterol and HDL-cholesterol in the HDRS group were increased to the peak point at postprandial 1·5, 2·5, 2·5, 1·5 and 1·5 h, those in the MDRS group were at postprandial 2·5, 3·5, 3·5, 3·5 and 3·5 h and those in the LDRS group were at postprandial 2·5, 3·5, 3·5, 1·5 and 3·5 h, respectively. The serum concentration of insulin in the HDRS group was higher (P < 0·05) than those in the MDRS group, and those in the MDRS group was also higher (P < 0·05) than those in the LDRS group at postprandial 0·5, 1·5 and 2·5 h, respectively. The serum concentrations of acetate, propionate and butyrate in the HDRS group were higher (P < 0·05) than those in the MDRS group, and those in the MDRS group were higher (P < 0·05) than in the LDRS group in each feeding cycle, in turn, respectively. The activity of fatty acid synthase (FAS) in the liver and abdominal adipose tissues, that of acetyl CoA carboxylase (ACC) in the myocardium and interscapular brown adipose tissues and that of the ATP-citrate lyase (ATP-CL) in the liver and interscapular brown adipose tissues in pigs of the HDRS group were higher (P < 0·05) than that of the MDRS group. The mRNA levels of FAS in the myocardium, liver and interscapular brown adipose tissues of pigs in the HDRS group were higher (P < 0·05) than those of the MDRS group. The activities and mRNA levels of FAS, ACC and ATP-CL in the myocardium, liver, abdominal and interscapular brown adipose tissues of the HDRS group were higher than those of the LDRS group. We conclude that the digestion rate of dietary starch affected not only the postprandial systemic circulating levels of glucose and insulin but also the lipid metabolism in weaned pigs. Dietary starch with higher digestion rate produces higher blood glucose and insulin response, ameliorates the blood lipid profiles and up-regulates the activity and gene expression profile of lipid metabolism-related genes in weaned pigs.

The xyn2 gene of a filamentous mesophilic fungus, Trichoderma reesei, coding xylanase 2 (Xyn2) was previously expressed in Pichia pastoris. In the present study, the recombinant Xyn2 was prepared from a 15 litre fermenter, and subsequently characterised. It has been confirmed to have a molecular mass of 21 kDa, an optimal pH of 6·0 and an optimal temperature of 60°C. When tested using oat-spelt xylan, it showed a Km and catalytic rate constant (kcat) of 1·1 mg/ml and 512·4/s, respectively. Analysis of the products from oat-spelt xylan degradation confirmed that the enzyme was an endoxylanase with xylotriose and xylobiose as the main degradation products. The unprocessed Xyn2 was supplemented to a xylan-containing diet to determine its influences on performance and nutrient digestibilities by weaned pigs. Results showed that the average body-weight gain increased 16·9 % when piglets received Xyn2 at a concentration of 500 U/kg diet. There also was a positive (0·05 < P < 0·10) effect on the digestibility values of crude protein, ash, Ca and acid-detergent fibre with Xyn2 supplementation. The potential benefits of Xyn2 in the nutrition of weaned pigs should make it an alternative applicant for industrial xylanase production.

This study evaluated whether arginine (Arg) supplementation could attenuate gut injury induced by Escherichia coli lipopolysaccharide (LPS) challenge through an anti-inflammatory role in weaned pigs. Pigs were allotted to four treatments including: (1) non-challenged control; (2) LPS-challenged control; (3) LPS+0·5 % Arg; (4) LPS+1·0 % Arg. On day 16, pigs were injected with LPS or sterile saline. At 6 h post-injection, pigs were killed for evaluation of small intestinal morphology and intestinal gene expression. Within 48 h of challenge, 0·5 % Arg alleviated the weight loss induced by LPS challenge (P = 0·025). In all three intestinal segments, 0·5 or 1·0 % Arg mitigated intestinal morphology impairment (e.g. lower villus height and higher crypt depth) induced by LPS challenge (P < 0·05), and alleviated the decrease of crypt cell proliferation and the increase of villus cell apoptosis after LPS challenge (P < 0·01). The 0·5 % Arg prevented the elevation of jejunal IL-6 mRNA abundance (P = 0·082), and jejunal (P = 0·030) and ileal (P = 0·039) TNF-α mRNA abundance induced by LPS challenge. The 1·0 % Arg alleviated the elevation of jejunal IL-6 mRNA abundance (P = 0·053) and jejunal TNF-α mRNA abundance (P = 0·003) induced by LPS challenge. The 0·5 % Arg increased PPARγ mRNA abundance in all three intestinal segments (P < 0·10), and 1·0 % Arg increased duodenal PPARγ mRNA abundance (P = 0·094). These results indicate that Arg supplementation has beneficial effects in alleviating gut mucosal injury induced by LPS challenge. Additionally, it is possible that the protective effects of Arg on the intestine are associated with decreasing the expression of intestinal pro-inflammatory cytokines through activating PPARγ expression.