Fecal Microbiota Transplantation Beneficially Regulates Intestinal Mucosal Autophagy and Alleviates Gut Barrier Injury.

Fecal microbiota transplantation (FMT) is one of the most effective ways to regulate the gut microbiota. Here, we investigated the effect of exogenous fecal microbiota on gut function from the perspective of analysis of the mucosal proteomes in a piglet model. A total of 289 differentially expressed proteins were annotated with 4,068 gene ontology (GO) function entries in the intestinal mucosa, and the levels of autophagy-related proteins in the forkhead box O (FoxO) signaling pathway were increased whereas the levels of proteins related to inflammation response were decreased in the recipient. Then, to assess the alleviation of epithelial injury in the Escherichia coli K88-infected piglets following FMT, intestinal microbiome-metabolome responses were determined. 16S rRNA gene sequencing showed that the abundances of beneficial bacteria, such as Lactobacillus and Succinivibrio, were increased whereas those of Enterobacteriaceae and Proteobacteria bacteria were decreased in the infected piglets following FMT. Metabolomic analysis revealed that levels of 58 metabolites, such as lactic acid and succinic acid, were enhanced in the intestinal lumen and that seven metabolic pathways, such as branched-chain amino acid metabolism pathways, were upregulated in the infected piglets following FMT. In concordance with the metabolome data, results of metagenomics prediction analysis also demonstrated that FMT modulated the metabolic functions of gut microbiota associated with linoleic acid metabolism. In addition, intestinal morphology was improved, a result that coincided with the decrease of intestinal permeability and the enhancement of mucins and mucosal expression of tight junction proteins in the recipient. Taken together, the results showed that FMT triggered intestinal mucosal protective autophagy and alleviated gut barrier injury through alteration of the gut microbial structure. IMPORTANCE The gut microbiota plays a crucial role in human and animal health, and its disorder causes multiple diseases. Over the past decade, FMT has gained increasing attention due to the success in treating Clostridium difficile infection (CDI) and inflammatory bowel disease (IBD). Although FMT appears to be effective, how FMT functions in the recipient remains unknown. Whether FMT exerts this beneficial effect through a series of changes in the host organism caused by alteration of gut microbial structure is also not known. In the present study, newborn piglets and E. coli K88-infected piglets were selected as models to explore the interplay between host and gut microbiota following FMT. Our results showed that FMT triggered intestinal mucosal autophagy and alleviated gut barrier injury caused by E. coli K88. This report provides a theoretical basis for the use of FMT as a viable therapeutic method for gut microbial regulation.

Chitosan-Zn Chelate Downregulates TLR4-NF-κB Signal Pathway of Inflammatory Response and Cell Death-Associated Proteins Compared to Inorganic Zinc.

The study was conducted to investigate the effect of chitosan-zinc chelate (CS-Zn) on TLR4-NF-κB signaling pathway and cell death-associated proteins in a weanling pig model. A total of 90 weaned piglets were allotted to three dietary treatments (the dietary treatments were as follows: (1) experimental diet with supplemental ZnSO4 (150 mg Zn/kg diet), (2) experimental diet with supplemental CS-Zn (150 mg Zn/kg diet), and (3) experimental diet with a supplemental mixture of chitosan and ZnSO4 (150 mg/kg Zn; the content of chitosan was equal to CS-Zn, which is according to molar basis)). The feeding trial lasted 30 days. The results showed that compared with ZnSO4 or CS+ZnSO4, CS-Zn decreased the expressions of the cell death-associated proteins Beclin-1, and Cleaved-Caspase3 and the ratio of LC3II/LC3I. The intestinal expressions of TLR4 and its downstream signals NF-κB, IKKß, and IκBα were down-regulated simultaneously. Moreover, the contents of pro-inflammatory cytokines IL-2, TNF-α, and IFN-γ were decreased. The results indicated that as organic zinc source, CS-Zn was more effective than ZnSO4 and the mixture of chitosan and ZnSO4 for inhibiting inflammatory response and decreasing the expressions of proteins associated with cell death. The great anti-inflammatory effect of CS-Zn was modulated by inhibiting the TLR4-NF-κB signaling pathway, and the effect of CS-Zn on down-regulating the expression of cell death-associated proteins might also closely be associated with the TLR4-NF-κB signaling pathway.

Effects of dietary fresh fermented soybean meal on growth performance, ammonia and particulate matter emissions, and nitrogen excretion in nursery piglets.

This study was conducted to investigate the effects of fresh fermented soybean meal (FSM) on the growth performance of nursery piglets, nitrogen excretion in feces, and the concentrations of ammonia (NH3) and particulate matter (PM) in the piggery. A total of 472 nursery piglets (Landrace×Yorkshire, (16.3±0.36) kg body weight) were randomly allocated into two treatments with 236 pigs in each treatment. The pigs were fed the basal diet without fresh FSM (control) or diet containing 10% (100 g/kg) fresh FSM (FSM group), and the crude protein content of the two groups was consistent. The feeding trial lasted for 28 d. The results showed that the pigs fed fresh FSM had increased (P<0.05) average daily gain (ADG) compared with the control. There was no significant difference (P>0.05) in feed to gain ratio (F:G) between the two groups. During the whole experiment, the concentration of NH3 in the piggery decreased (P<0.05) by 19.0%, and the concentrations of PM (PM10 and PM2.5) in the piggery decreased (P<0.05) by 19.9% and 11.6%, respectively, in the FSM group, compared with the control. The ammonia nitrogen and nitrite content in feces increased (P<0.05) by 32.9% and 28.4%, respectively, in the FSM group. The fecal pH declined (P<0.05) significantly in the FSM group compared with the control. At the end of experiment, total protein (TP) concentration was increased (P<0.05) significantly and blood urea nitrogen (BUN) concentration was decreased (P<0.05) for pigs fed the diet with fresh FSM. The results indicated that dietary fresh FSM not only improved the growth performance of nursery piglets, but also reduced the NH3 concentration in the piggery due to nitrogen conversion, and decreased the concentrations of PM10 and PM2.5 in the piggery.

Changes of gut microbiota structure and morphology in weaned piglets treated with fresh fermented soybean meal.

This study investigated the effects of dietary fresh fermented soybean meal (FSM) on the intestinal microbiota and metabolites, bacterial enzyme activity and intestinal morphology of weaning piglets. A total of 64 weaned piglets were randomly allocated into two treatments. A corn-soybean-based diet was used as the control and other treatment was fed the same basal diet containing 15% fresh FSM. The feeding trial lasted for 21 days. Bacterial community structure and diversity in the cecum and colon were assessed using pyrosequencing-based analysis. The results showed that the phylum level, Firmicutes, Bacteroidetes, Proteobacteria and Tenericutes were dominant in the cecum or colon. Gut Firmicutes increased, while Bacteroidetes and Proteobacteria decreased in the fresh FSM-fed piglets. At the genus level, the relative abundances of butyrate-producing bacteria, Lactobacillus and Prevotella were higher in both cecum and colon of fresh FSM fed piglets. Meanwhile, fresh FSM could promote the development of intestinal morphological and reduce the incidence of diarrhea. The results indicated that fresh FSM might change intestinal function by influencing intestinal microenvironment.

Exogenous Fecal Microbiota Transplantation from Local Adult Pigs to Crossbred Newborn Piglets.

This study was conducted to investigate the effect of exogenous fecal microbiota transplantation on gut bacterial community structure, gut barrier and growth performance in recipient piglets. Twelve litters of Duroc × Landrace × Yorkshire piglets of the same birth and parity were weighed and divided into two groups. One group (recipient piglets) was inoculated orally with fecal microbiota suspension of healthy adult Jinhua pigs daily from day 1 to day 11. The other (control) was given orally the same volume of sterile physiological saline at the same time. The experiment lasted 27 days. The results showed that the relative abundance of Firmicutes, Prevotellaceae, Lachnospiraceae, Ruminococcus, Prevotella, and Oscillospira in the colon of recipient piglets was increased. Proteobacteria, Fusobacteriaceae, Clostridiaceae, Pasteuriaceae, Alcaligenaceae, Bacteroidaceae, Veillonellaceae, Sutterella, Escherichia, and Bacteroides in the colon of recipient piglets were decreased. An average daily weight gain of recipient piglets was increased, and diarrhea incidence of the recipient was decreased during the trial. Intestinal morphology and tight junction barrier of recipient piglets were improved. The optical density of sIgA+ cells, the number of goblet cells and relative expressions of MUC2 in the intestinal mucosa of recipient piglets were enhanced. Protein expressions of ß-defensin 2 and mRNA expressions of TLR2 and TLR4 in the intestinal mucosa of recipient piglets were also increased. These findings supported that the exogenous fecal microbiota had significant effects on animal's growth performance, intestinal barrier function, and innate immune via modulating the composition of the gut microbiota.

The Expression of Zinc Transporters Changed in the Intestine of Weaned Pigs Exposed to Zinc Chitosan Chelate.

This study was conducted to investigate the effect of zinc chitosan chelate (CS-Zn) on zinc transporter expression and content of tissue zinc in weaned piglets. A total of 90 weaned pigs (Duroc × Landrace × Yorkshire) were randomly allocated to treatment groups with supplementation of 100 mg/kg zinc as ZnSO4, 100 mg/kg zinc as mixture of ZnSO4 and chitosan, or 100 mg/kg zinc as CS-Zn, respectively. After 30 days of trial, 18 piglets (six pigs per treatment) were killed and the samples of duodenal mucosa were taken for analysis of zinc transporter mRNA expressions and protein abundance. The results show that CS-Zn more effectively increases (p < 0.05) the average daily gain (ADG) and serum zinc concentration. Zinc concentration in the liver and kidney did not differ between treatments. The mRNA expressions of ZnT1, ZIP4, and ZIP5 in CS-Zn treatment were all upregulated (p < 0.05) than ZnSO4 or mixture of ZnSO4 and chitosan groups. ZnT1 abundance was greater (p < 0.05) with CS-Zn as compared with ZnSO4 and mixture of ZnSO4 and chitosan treatments, whereas ZIP4 and ZIP5 abundance was higher (p < 0.05) in ZnSO4 group. The results indicate that CS-Zn is more effective in serum zinc accumulation, and it might regulate zinc homeostasis by affecting zinc transporter mRNA expression and absorption mechanism might be different with ZnSO4.

Changes in diarrhea, nutrients apparent digestibility, digestive enzyme activities of weaned piglets in response to chitosan-zinc chelate.

A total of 120 weanling barrows weighing 6.11 ± 0.20 kg were randomly allotted to four treatments with three replications (i.e. pen) of ten piglets per replicate. Pigs were received corn-soybean basal diet (control) or the same basal diet supplemented with the following sources of zinc: (i) 100 mg/kg of Zn as ZnSO4 ; (ii) 100 mg/kg of Zn as chitosan-Zn chelate (CS-Zn); and (iii) 100 mg/kg of Zn as ZnSO4 mixed with chitosan (CS + ZnSO4 ). The results showed that CS-Zn could highly improve average daily gain and average daily feed intake than those of ZnSO4 or CS+ ZnSO4 (P < 0.05). The pigs fed dietary CS-Zn had lower diarrhea incidence and higher apparent digestibility of crude protein than those of the pigs fed dietary ZnSO4 (P < 0.05). The protease activities in duodenal content of the pigs receiving CS-Zn diets was higher than that of the pigs fed dietary ZnSO4 or CS + ZnSO4 (P < 0.05). The amylase activity in duodenal content of the pigs fed dietary CS-Zn was higher than that of the pigs receiving ZnSO4 diets or basal diets (P < 0.05). These results indicated that dietary CS-Zn showed different bioactivities from ZnSO4 or CS + ZnSO4 in reducing the incidence of diarrhea, improving activities of digestive enzymes and growth performance of weaned pigs.