Impacts of Fructose on Intestinal Barrier Function, Inflammation and Microbiota in a Piglet Model.

The metabolic disorder caused by excessive fructose intake was reported extensively and often accompanied by intestinal barrier dysfunction. And the rising dietary fructose was consumed at an early age of human. However, related researches were almost conducted in rodent models, while in the anatomy and physiology of gastrointestinal tract, pig is more similar to human beings than rodents. Hence, weaned piglets were chosen as the model animals in our study to investigate the fructose's impacts on intestinal tight junction, inflammation response and microbiota structure of piglets. Herein, growth performance, inflammatory response, oxidation resistance and ileal and colonic microbiota of piglet were detected after 35-day fructose supplementation. Our results showed decreased tight junction gene expressions in piglets after fructose addition, with no obvious changes in the growth performance, antioxidant resistance and inflammatory response. Moreover, fructose supplementation differently modified the microbiota structures in ileum and colon. In ileum, the proportions of Streptococcus and Faecalibacterium were higher in Fru group (fructose supplementation). In colon, the proportions of Blautia and Clostridium sensu stricto 1 were higher in Fru group. All the results suggested that tight junction dysfunction might be an earlier fructose-induced event than inflammatory response and oxidant stress and that altered microbes in ileum and colon might be the potential candidates to alleviate fructose-induced intestinal permeability alteration.

Grape Seed Proanthocyanidin Affects Lipid Metabolism via Changing Gut Microflora and Enhancing Propionate Production in Weaned Pigs.

BACKGROUND: It is not clear whether dietary grape seed proanthocyanidin (GSP) affects mammalian lipid metabolism via the gut microbiota. OBJECTIVE: The aim of this study was to evaluate the contribution of the gut microbiota to the effect of dietary GSP. METHODS: This study was divided into 3 separate experiments using Duroc × Landrace × Yorkshire pigs (50% male) weaned at day 28 and then fed the same basal diet (NC). In Experiment 1, 90 pigs were fed NC or NC with 250 mg GSP/kg (GSP) or 400 mg betaine/kg [positive control (PC)] for 28 d. In Experiment 2, 30 pigs were fed NC, GSP, or GSP with antibiotics (GSP + Abx) diets for 14 d. In Experiment 3, pigs were fed NC, NC plus 1 g sodium propionate/kg (SP), or NC plus 1 g sodium butyrate/kg (SB) diet for 14 d. Serum biochemical indexes, SCFA concentrations, and microbial composition were determined. RESULTS: In Experiment 1, compared with the GSP group, visceral adipocyte area was higher in the NC (28.6%) and PC (18.2%) groups (P ≤ 0.05). Colonic propionate and butyrate concentrations were 30.2% and 3.6% higher in the GSP group than in the NC group, respectively (P ≤ 0.05). In Experiment 2, compared with the GSP group, the NC group had a 108% higher Firmicutes to Bacteroidetes ratio and had 50.4%, 61.2%, and 82.3% lower abundance of Akkermansia, Alistipes, and Bacteroides, respectively (P ≤ 0.05); antibiotics removed these effects of GSP. In Experiment 3, serum peptide YY was 19.5% higher in the SP group than in the NC group (P ≤ 0.05), and it did not differ between the SB and NC groups (P > 0.05). CONCLUSIONS: GSP affected lipid metabolism in weaned pigs, which is associated with changed gut microbiota and enhanced microbial propionate production. These findings provide potential mechanisms for GSP intake to improve lipid metabolism.

Moderate Dietary Protein Restriction Optimized Gut Microbiota and Mucosal Barrier in Growing Pig Model.

Appropriate protein concentration is essential for animal at certain stage. This study evaluated the effects of different percentages of dietary protein restriction on intestinal health of growing pigs. Eighteen barrows were randomly assigned to a normal (18%), low (15%), and extremely low (12%) dietary protein concentration group for 30 days. Intestinal morphology and permeability, bacterial communities, expressions, and distributions of intestinal tight junction proteins, expressions of biomarkers of intestinal stem cells (ISCs) and chymous bacterial metabolites in ileum and colon were detected. The richness and diversity of bacterial community analysis with Chao and Shannon index were highest in the ileum of the 15% crude protein (CP) group. Ileal abundances of Streptococcaceae and Enterobacteriaceae decreased respectively, while beneficial Lactobacillaceae, Clostridiaceae_1, Actinomycetaceae, and Micrococcaceae increased their proportions with a protein reduction of 3 percentage points. Colonic abundances of Ruminococcaceae, Christensenellaceae, Clostridiaceae_1, Spirochaetaceae, and Bacterodales_S24-7_group declined respectively, while proportions of Lachnospiraceae, Prevotellaceae, and Veillonellaceae increased with dietary protein reduction. Concentrations of most bacterial metabolites decreased with decreasing dietary protein concentration. Ileal barrier function reflected by expressions of tight junction proteins (occludin, zo-3, claudin-3, and claudin-7) did not show significant decrease in the 15% CP group while sharply reduced in the 12% CP group compared to that in the 18% CP group. And in the 15% CP group, ileal distribution of claudin-3 mainly located in the cell membrane with complete morphological structure. In low-protein treatments, developments of intestinal villi and crypts were insufficient. The intestinal permeability reflected by serous lipopolysaccharide (LPS) kept stable in the 15% CP group while increased significantly in the 12% CP group. The expression of ISCs marked by Lgr5 slightly increased in ileum of the 15% CP group. Colonic expressions of tight junction proteins declined in extremely low protein levels. In conclusion, moderate protein restriction (15% CP) can optimize the ileal microbiota structure via strengthening beneficial microbial populations and suppressing harmful bacterial growth and altering the function of ileal tight junction proteins as well as epithelial cell proliferation.

Moderate dietary protein restriction alters the composition of gut microbiota and improves ileal barrier function in adult pig model.

This study was conducted to investigate impacts of dietary protein levels on gut bacterial community and gut barrier. The intestinal microbiota of finishing pigs, fed with 16%, 13% and 10% crude protein (CP) in diets, respectively, were investigated using Illumina MiSeq sequencing. The ileal bacterial richness tended to decrease when the dietary protein concentration reduced from 16% to 10%. The proportion of Clostridium_sensu_stricto_1 in ileum significantly decreased, whereas Escherichia-Shigella increased with reduction of protein concentration. In colon, the proportion of Clostridium_sensu_stricto_1 and Turicibacter increased, while the proportion of RC9_gut_group significantly decreased with the dietary protein reduction. Notably, the proportion of Peptostreptococcaceae was higher in both ileum and colon of 13% CP group. As for metabolites, the intestinal concentrations of SCFAs and biogenic amines decreased with the dietary protein reduction. The 10% CP dietary treatment damaged ileal mucosal morphology, and decreased the expression of biomarks of intestinal cells (Lgr5 and Bmi1), whereas the expression of tight junction proteins (occludin and claudin) in 13% CP group were higher than the other two groups. In conclusion, moderate dietary protein restriction (13% CP) could alter the bacterial community and metabolites, promote colonization of beneficial bacteria in both ileum and colon, and improve gut barrier function.

Supplementation of Milky Flavors Improves the Reproductive Performance and Gut Function Using Sow Model.

This study was conducted to evaluate the effect of flavors on reproductive performance of sows and we also studied its effect on gut barrier function. Forty-eight Landrace × Yarkshire sows were randomly allotted and fed a basal diet added 0%, 0.05% or 0.10% flavor feed, respectively from parturition to day 28 of weaning. The results showed that supplementation of 0.05% or 0.10% flavors increased average daily feed intake (ADFI) of sows and average daily gain (ADG) of piglets, decreased the weight losses of sows, increased the survival ratio of weaning piglets (P < 0.05), especially shorten the post-weaning estrus interval significantly (P < 0.05). Supplementation of flavor additives tend to reduce the weight losses of sows and raise the survival ratio of piglet weaned (P > 0.05). Moreover, addition of flavors in diets reduced the intestinal permeability and enhanced digestibility of dry matter, crude protein, and energy (P < 0.05). Flavors supplementation significantly increased the level of gonadotropin releasing hormne (GnRH) of serum in sows after weaning. In conclusion, the results suggested that supplementation of dietary flavors could improve digestibility of nutrients and the reproductive performance of sows as well as the gut barrier function.

Roles of Biogenic Amines in Intestinal Signaling.

Biogenic amines in the gastrointestinal tract are important metabolites of dietary protein and amino acids with the help of gut digestive enzymes and microbes, which play a crucial role in the regulation of intestinal functions, including digestion, absorption, and local immunity. However, high concentrations of biogenic amines can induce adverse reactions and are harmful to animal's health. Therefore, it is crucial to have a clear understanding of how different biogenic amines interact with a body's intestinal function signaling pathways and to monitor the content of biogenic amines in the gastrointestinal tract. And in turn, the proper concentration of dietary protein and balanced amino acids for humans and livestock could be given. Though numerous methods have been developed and improved for the detection of biogenic amines in foods or wines much less attention has been paid directly to the determination of amine levels in the gastrointestinal tract. In this article, we mainly focus on the interaction of amines with the intestinal function signaling pathway and the broad impacts on animal physiology, and our modified method to accurately and quickly detect the biogenic amines in the digesta of an animal intestine.

Dietary grape seed proanthocyanidins (GSPs) improve weaned intestinal microbiota and mucosal barrier using a piglet model.

Proanthocyanidins have been suggested as an effective antibiotic alternative, however their mechanisms are still unknown. The present study investigated the effects of grape seed proanthocyanidins on gut microbiota and mucosal barrier using a weaned piglet model in comparison with colistin. Piglets weaned at 28 day were randomly assigned to four groups treated with a control ration, or supplemented with 250 mg/kg proanthocyanidins, kitasamycin/colistin, or 250 mg/kg proanthocyanidins and half-dose antibiotics, respectively. On day 28, the gut chyme and tissue samples were collected to test intestinal microbiota and barrier function, respectively. Proanthocyanidins treated piglets had better growth performance and reduced diarrhea incidence (P < 0.05), accompanied with decreased intestinal permeability and improved mucosal morphology. Gene sequencing analysis of 16S rRNA revealed that dietary proanthocyanidins improved the microbial diversity in ileal and colonic digesta, and the most abundant OTUs belong to Firmicutes and Bacteroidetes spp.. Proanthocyanidins treatment decreased the abundance of Lactobacillaceae, and increased the abundance of Clostridiaceae in both ileal and colonic lumen, which suggests that proanthocyanidins treatment changed the bacterial composition and distribution. Administration of proanthocyanidins increased the concentration of propionic acid and butyric acid in the ileum and colon, which may activate the expression of GPR41. In addition, dietary proanthocyanidins improved the antioxidant indices in serum and intestinal mucosa, accompanied with increasing expression of barrier occludin. Our findings indicated that proanthocyanidins with half-dose colistin was equivalent to the antibiotic treatment and assisted weaned animals in resisting intestinal oxidative stress by increasing diversity and improving balance of gut microbes.

Dietary Sodium Butyrate Decreases Postweaning Diarrhea by Modulating Intestinal Permeability and Changing the Bacterial Communities in Weaned Piglets.

BACKGROUND: The vast majority of substances used as alternatives to antibiotics produce inconsistent results and rarely equal the effectiveness of in-feed antibiotics. OBJECTIVE: This study evaluated the effects of the combined use of sodium butyrate (SB) and reduced antibiotics in a piglet diet in promoting performance and to control weaning diarrhea. METHODS: Piglets weaned at 28 d were randomly assigned to a corn-soybean meal control ration [negative control (NC)]; a similar ration with 50 mg kitasamycin/kg, 20 mg colistin sulfate/kg, and 1000 mg encapsulated SB/kg [reduced antibiotics + SB (ASB)]; or to a ration with 100 mg kitasamycin/kg and 40 mg colistin sulfate/kg [positive control (PC)] for 28 d. Performance, diarrhea incidence, intestinal permeability, and changes in the bacterial communities in the ileum and colon were determined. RESULTS: Weight gain and the ratio of weight gain to feed intake were significantly greater in the ASB and PC piglets than in the NC piglets (P < 0.05). Diarrhea incidence was lower in the ASB and PC piglets than in the NC piglets (P < 0.05). Urinary lactulose to mannitol ratios were 25% and 30% lower, respectively, whereas jejunal and colonic occludin protein expressions were significantly greater in the ASB and PC piglets compared with the NC piglets (P < 0.05). In the intestinal mucosa, malondialdehyde was lower in the ASB and PC piglets (by 42% and 43%, respectively), whereas tumor necrosis factor α (TNF-α) was 63% lower in the ASB piglets and 59% lower in the PC piglets compared with the NC piglets (P < 0.05). 16S ribosomal RNA gene sequence analysis revealed a higher colonic Shannon index and a lower colonic Simpson index in the ASB and PC piglets than in the NC piglets. In addition, the ASB and PC treatments caused a striking decrease in Lactobacillaceae and a noticeable increase in Clostridiaceae in the ileal and colonic lumen, as well as increases in Ruminococcaceae, Lachnospiraceae, and Bacteroidetes in the colonic lumen. CONCLUSION: Collectively, our results support an important role for SB in improving performance and decreasing diarrhea incidence in weaned piglets by modulation of intestinal permeability and the bacterial communities in the ileum and colon.

Effects of particle size and drying methods of corn on growth performance, digestibility and haematological and immunological characteristics of weaned piglets.

The objective of this study was to evaluate the effects of particle size and drying methods of corn on growth performance of weaned piglets. Crossbreed weaned piglets (n = 192; Duroc × Landrace × Large White) were assigned to one of four treatments (2 × 2 factorial arrangement). All piglets were fed corn-soybean meal diets and treatments were (1) hot air-dried and coarsely ground corn, (2) hot air-dried and finely ground corn, (3) sun-dried and coarsely ground corn and (4) sun-dried and finely ground corn. The results showed that finely ground corn (FGC) improved the performance of piglets. Additionally, the apparent total tract digestibility (ATTD) of gross energy (GE) and ether extract (EE) were increased by FGC, but the drying methods did not affect the performance of piglets or ATTD. Furthermore, smaller particle size significantly decreased the intestinal permeability, which was also not influenced by drying methods. FGC increased the total number of white blood cells, but not other blood parameters. Finally, the level of serum interleukin-1 was decreased by fine grinding and that of serum tumour necrosis factor α was decreased by sun drying. Conversely, these characteristics of weaned piglets can hardly have been affected either by the corn drying method or its interaction with grinding methods.

Determination of glycinin in soybean and soybean products using a sandwich enzyme-linked immunosorbent assay.

This study performs a sandwich ELISA for detection of trace amounts of glycinin in soybean products. We designed a soy-free mouse model to produce anti-glycinin monoclonal antibodies with high affinity and specificity. Using the monoclonal antibody as coating antibody, with the rabbit anti-glycinin polyclonal antibody as a detected antibody, the established sandwich ELISA showed high specificity for glycinin with minimum cross-reactions with other soy proteins. The practical working range of the determination was 3-200 ng/mL with detection limit of 1.63 ng/mL. The regaining of glycinin in spiked soybean samples were between 93.8% and 103.3% with relative standard deviation less than 8.3% (intra-day) and 10.5% (inter-day). The developed assay was used in analysing 469 soybean samples and five soybean products under different processing. The assay provides a specific and sensitive method for screening of glycinin and allows for further investigation into hypersensitive mechanisms to soybean proteins.

Dietary grape-seed procyanidins decreased postweaning diarrhea by modulating intestinal permeability and suppressing oxidative stress in rats.

This study was conducted to evaluate the effects of grape-seed procyanidins in controlling weaning diarrhea using a rat model. Weaned rats were fed either the basal diet or basal diet supplemented with either 250 mg/kg grape-seed procyanidins or 2000 mg/kg ZnO. Treated rats had better performance with a reduced incidence of diarrhea (P < 0.05). Both ZnO and grape-seed procyanidins significantly reduced urinary lactulose to mannitol ratios (P < 0.05) and enhanced the mRNA and protein expression of the intestinal mucosal tight junction proteins Ocln/ZO-1 (P < 0.05). Grape-seed procyanidins increased the activities of SOD, GSH-Px, and GSH while decreasing the level of MDA in the intestinal mucosa (P < 0.05). Furthermore, an in vitro investigation revealed that supplementation with grape-seed procyanidins in IEC-6 intestinal epithelial cells significantly enhanced the expression of Ocln/ZO-1 under H(2)O(2)-induced oxidative stress. Collectively, these results indicate that grape-seed procyanidins have the potential to prevent weaning diarrhea by reducing intestinal permeability and improving antioxidant indices.