This study examined effects of dietary protein sources and levels on intestinal health of 21 to 35 d-old weaned piglets fed antibiotics-free diets. A total of 150 weaned piglets (21 d of age) were allotted to 5 dietary treatment groups. Diets were formulated, based on corn-soybean meal, with different protein sources (fish meal and soy protein concentrate) to provide different dietary CP levels. Piglets within 5 dietary treatments were fed diets as follows, respectively: 1) control diet of 17% CP (control); 2) 19% CP diets formulated with more soy protein concentrate (SPC19); 3) fish meal (FM19); 4) 23.7% CP diets formulated with more soy protein concentrate (SPC23); 5) fish meal (FM23). The results showed that piglets from control group had higher ADG and lower incidence of diarrhea compared with those of other groups (P < 0.05). The incidence of diarrhea of piglets in FM19 group was lower than those from SPC23 group and FM23 group (P < 0.05). With the higher CP levels, villous height and villous height to crypt depth ratio of piglets in the duodenum and jejunum were decreased (P < 0.05), but crypt depth was increased (P < 0.05). Comparing control group and other groups, we found the expression of inflammatory cytokines interleukin-1β (IL-1β) and interferon-γ (IFN-γ) were increased (P < 0.05) in the jejunum and colon of piglets, as did cystic fibrosis transmembrane conductance regulators (CFTR) in the distal colon. The relative transcript abundance of Zonula occludens-1 (ZO-1) in the jejunum, and occludin in the jejunum and ileum of piglets fed 23.7% CP diets were reduced compared with those fed control diet (P < 0.05). In conclusion, the 17% CP diet without in-feed antibiotics helped improve growth performance and relief of diarrhea of 21 to 35 d-old weaned piglets. Dietary CP level, rather than its source (either fish meal or soy protein concentrate), has more significant impacts on the growth performance and intestinal health of 21 to 35 d-old weaned piglets when fed antibiotics-free diets.

The objective of this study was to investigate the effects of Lactobacillus reuteri LR1, a new strain isolated from the feces of weaned pigs, on the growth performance, intestinal morphology, immune responses, and intestinal barrier function in weaned pigs. A total of 144 weaned pigs (Duroc × Landrace × Yorkshire, 21 d of age) with an initial BW of 6.49 ± 0.02 kg were randomly assigned to 3 dietary treatments with 8 replicate pens, each of per treatment and 6 pigs. Pigs were fed a basal diet (CON, controls), the basal diet supplemented with 100 mg/kg olaquindox and 75 mg/kg aureomycin (OA) or the basal diet supplemented with 5 × 1010 cfu/kg L. reuteri LR1 for a 14-d period. At the end of study, the ADG, ADFI, and G:F were calculated, and 1 randomly selected pig from each pen was euthanized for sample collection. The LR1 increased ADG (22.73%, P < 0.05) compared with CON. The villus height of the ileum was increased (P < 0.05) and crypt depth in duodenum was reduced (P < 0.05), along with increased (P < 0.05) villus height to crypt depth ratio of the jejunum and ileum by LR1 compared with CON and OA. LR1 increased (P < 0.05) ileal mucosal content of IL-22 and transforming growth factor-β compared with OA. Compared with CON, LR1 increased (P < 0.05) and OA decreased (P < 0.05) the ileal content of secretory immunoglobulin A (sIgA), and the abundance of transcripts of porcine β-defensin 2 and protegrin 1-5. Compared with CON, LR1 increased (P < 0.05) tight junction protein zonula occludens-1 and occludin transcripts in the mucosa of the jejunum and ileum, and those of mucin-2 in ileal mucosa. The relative expression of toll-like receptor 2 (TLR2) and TLR4 were increased (P < 0.05) in ileal mucosa in pigs fed LR1 compared with CON. In conclusion, these data indicated that dietary LR1 supplementation at 5 × 1010 cfu/kg improved growth performance, intestinal morphology, and intestinal barrier function in weaned pigs.

Intestinal epithelial barrier damage disrupts immune homeostasis and leads to many intestinal disorders. Lactobacillus reuteri strains have probiotic functions in their modulation of the microbiota and immune system in intestines. In this study, the effects of L. reuteri LR1, a new strain isolated from the feces of weaning piglets, on intestinal epithelial barrier damage in IPEC-1 cells caused by challenge with enterotoxigenic Escherichia coli (ETEC) K88 were examined. It was found that L. reuteri LR1, in large part, offset the ETEC K88-induced increase in permeability of IPEC-1 cell monolayers and decreased the adhesion and invasion of the coliform in IPEC-1 cells. In addition, L. reuteri LR1 increased transcript abundance and protein contents of tight junction (TJ) proteins zonula occluden-1 (ZO-1) and occludin in ETEC K88-infected IPEC-1 cells, whereas it had no effects on claudin-1 and F-actin expression. Using colloidal gold immunoelectron microscopy, these effects of L. reuteri LR1 on ZO-1 and occludin content in IPEC-1 cells were confirmed. By using ML-7, a selective inhibitor of myosin light-chain kinase (MLCK), the beneficial effect of L. reuteri LR1 on contents of ZO-1 and occludin was shown to be dependent on the MLCK pathway. In conclusion, L. reuteri LR1 had beneficial effects on epithelial barrier function consistent with increasing ZO-1 and occludin expression via a MLCK-dependent manner in IPEC-1 cells during challenge with ETEC K88.

Lactating mammary glands are among the most active lipogenic organs and provide a large percentage of bioactive lipids and calories for infant growth. The branched-chain amino acid (BCAA) valine is known to modulate fatty acids synthesis in adipose tissue; however, its effects on fat metabolism and the underlying mechanisms in mammary glands remain to be determined. Valine supplementation during late pregnancy significantly increased the contents of total milk fat, triglyceride, sphingomyelin, and polyunsaturated fatty acids in the colostrum of gilts. Further study in porcine mammary epithelial cells (PMECs) confirmed that valine upregulated the phosphorylation levels of AKT-activated MTOR and subsequently induced the nuclear accumulation of sterol regulatory element binding protein 1 (SREBP1), thus increasing the expression of proteins related to fatty acids synthesis and intracellular triacylglycerol content. Inhibition of AKT/MTOR signaling or silencing of SREBP1 in PMECs downregulates the expression of proteins related to fatty acids synthesis and intracellular triacylglycerol content. Our findings indicated that valine enhanced milk fat synthesis of colostrum in porcine mammary glands via the AKT/MTOR/SREBP1 signaling pathway.

This experiment studied the effects of dietary protein sources and levels on the gut health of piglets, pH value, and concentrations of microbial metabolites (ammonia-N, volatile fatty acids [VFA], and polyamines) in the distal colonic and proximal colonic digesta of piglets weaned at 21 d of age. A total of 150 early-weaned piglets were allotted randomly to 5 diets: 1) control diet (CT; 17% CP), 2) CT formulated with more soy protein concentrate (SPC19; 19% CP), 3) more fish meal (FM19; 19% CP), 4) CT formulated with more soy protein concentrate (SPC23; 23% CP), and 5) more fish meal (FM23; 23%CP). Results showed high protein level increased fecal score (P < 0.05), but different protein sources did not (P > 0.05). The pH value and ammonia-N concentration of digesta in the proximal and distal colon of FM23 were significantly higher (P < 0.05) than those of CT. Acetic acid, propionic acid, butyric acid and valeric acid concentrations in the proximal colon of FM23 exceeded those of CT, SPC19, and FM19 (P < 0.05); however, isobutyric acid and isovaleric acid were not affected (P > 0.05). Histamine and spermidine concentrations of FM23 were higher than those of other treatments (P < 0.05). Propionic acid and butyric acid concentrations in the distal colon were higher of FM23 than of FM19 (P < 0.05); putrescine, histamine and spermidine were higher of FM23 than of LP and FM19 (P < 0.05). It was concluded that high dietary CP content increased microbial metabolites (ammonia-N, histamine, putrescine) in colonic digesta and aggravated piglets' diarrhea.