Dietary supplementation of benzoic acid and essential oils combination enhances intestinal resilience against LPS stimulation in weaned piglets

Background The benefits of combining benzoic acid and essential oils(BAO)to mitigate intestinal impairment during the weaning process have been well established,while the detailed underlying mechanism has not been fully elucidated.Previous research has primarily focused on the reparative effects of BAO on intestinal injury,while neglecting its potential in enhancing intestinal stress resistance.Methods In this study,we investigated the pre-protective effect of BAO against LPS-induced stress using a modified experimental procedure.Piglets were pre-supplemented with BAO for 14 d,followed by a challenge with LPS or saline to collect blood and intestinal samples.Results Our findings demonstrated that BAO supplementation led to significant improvements in piglets’final weight,average daily gain,and feed intake/body gain ratio.Additionally,BAO supplementation positively influenced the composition of intestinal microbiota,increasing beneficial Actinobacteriota and Alloprevotella while reducing harmful Desulfobacterota,Prevotella and Oscillospira.Furthermore,BAO supplementation effectively mitigated oxidative disturbances and inflammatory responses induced by acute LPS challenge.This was evidenced by elevated levels of T-AOC,SOD,and GSH,as well as decreased levels of MDA,TNF-α,and IL-6 in the plasma.Moreover,piglets subjected to LPS challenge and pre-supplemented with BAO exhibited significant improvements in intestinal morphological structure and enhanced integrity,as indicated by restored expression levels of Occludin and Claudin-1 compared to the non-supplemented counterparts.Further analysis revealed that BAO supplementation enhanced the jejunal antioxidative capacity by increasing GSH-Px levels and decreasing MDA levels under the LPS challenge and stimulated the activation of the Nrf2 signaling pathway.Additionally,the reduction of TLR4/NF-κB/MAPK signaling pathways activation and proinflammatory factor were also observed in the jejunal of those piglets fed with BAO.Conclusions In summary,our study demonstrates that pre-supplementation of BAO enhances the anti-stress capacity of weaned piglets by improving intestinal microbiota composition,reinforcing the intestinal barrier,and enhancing antioxidative and anti-inflammatory capabilities.These effects are closely associated with the activation of Nrf2 and TLR4/NF-κB/MAPK signaling pathways.

Dietary fibers with low hydration properties exacerbate diarrhea and impair intestinal health and nutrient digestibility in weaned piglets

Background:This study aimed to investigate the hydration properties of different-source fibrous materials by com-paring their water-binding capacity(WBC),water swelling capacity(WSC),viscosity,and in vivo effects of selected samples on growth performance,nutrient digestibility,diarrhea,and intestinal health in weaned piglets.Methods:A total of 13 commercially available fibrous materials were first compared in chemical composition and in vitro hydration property.Subsequently,40 weaned piglets were randomized to five experimental dietary groups(8 piglets per group):control diet(a basal diet without dietary fiber,CON),basal diet supplemented with 5%microcrys-talline cellulose(MCC),5%wheat bran(WB),5%Moringa oleifera leaf powder(MOLP),or 5%sugar beet pulp(SBP),fol-lowed by analyzing their growth performance and diarrhea rate in a 28-d experiment.After the feeding experiment,anaesthetized piglets were killed,and their intestinal and colon content or plasma samples were analyzed in nutrient digestibility,intestinal morphology,intestinal barrier,short-chain fatty acids(SCFAs),and bacterial population.Results:In vitro studies showed low hydration properties for WB and MCC,while medium hydration properties for MOLP and SBP.In vivo studies indicated that compared with medium hydration property groups,low hydration prop-erty groups showed(1)exacerbated diarrhea,impaired intestinal health,and reduced apparent fecal digestibility of dry matter,gross energy,acid detergent fiber,and neutral detergent fiber;(2)decreased SCFAs concentration and rela-tive levels of Lactobacillus and Bifidobacterium,but increased levels of Escherichia coli and Brachyspira hyodysenteriae in colon contents.Additionally,SBP showed optimal performance in reducing diarrhea and increasing SCFAs produc-tion.Correlation analysis revealed a positive correlation of fiber hydration properties with in vitro SCFAs production,and diarrhea index and nutrient digestibility were negatively and positively correlated with SCFAs levels in the colon contents of weaned piglets,respectively.Conclusions:Different-source dietary fibers varied in their hydration properties and impacts on diarrhea,microbial composition and SCFAs production in weaned piglets.WB and MCC could exacerbate diarrhea and impair nutrient digestibility,probably because their low hydration properties were detrimental to gut microbial homeostasis and fermentation.Our findings provide new ideas for rational use of fiber resources in weaned piglets.

Effects of lactic acid bacteria-fermented formula milk supplementation on ileal microbiota,transcriptomic profile,and mucosal immunity in weaned piglets

Background:Lactic acid bacteria(LAB)participating in milk fermentation naturally release and enrich the fermented dairy product with a broad range of bioactive metabolites,which has numerous roles in the intestinal health-promot-ing of the consumer.However,information is lacking regarding the application prospect of LAB fermented milk in the animal industry.This study investigated the effects of lactic acid bacteria-fermented formula milk(LFM)on the growth performance,intestinal immunity,microbiota composition,and transcriptomic responses in weaned piglets.A total of 24 male weaned piglets were randomly divided into the control(CON)and LFM groups.Each group consisted of 6 replicates(cages)with 2 piglets per cage.Each piglet in the LFM group were supplemented with 80 mL LFM three times a day,while the CON group was treated with the same amount of drinking water.Results:LFM significantly increased the average daily gain of piglets over the entire 14 d(P<0.01)and the average daily feed intake from 7 to 14 d(P<0.05).Compared to the CON group,ileal goblet cell count,villus-crypt ratio,sIgA,and lactate concentrations in the LFM group were significantly increased(P<0.05).Transcriptomic analysis of ileal mucosa identified 487 differentially expressed genes(DEGs)between two groups.Especially,DEGs involved in the intestinal immune network for IgA production pathways,such as polymeric immunoglobulin receptor(PIGR),were significantly up-regulated(P<0.01)by LFM supplementation.Moreover,trefoil factor 2(TFF2)in the LFM group,one of the DEGs involved in the secretory function of goblet cells,was also significantly up-regulated(P<0.01).Sequenc-ing of the 16S rRNA gene of microbiota demonstrated that LFM led to selective enrichment of lactate-producing and short-chain fatty acid(SCFA)-producing bacteria in the ileum,such as an increase in the relative abundance of Entero-coccus(P=0.09)and Acetitomaculum(P<0.05).Conclusions:LFM can improve intestinal health and immune tolerance,thus enhancing the growth performance of weaned piglets.The changes in microbiota and metabolites induced by LFM might mediate the regulation of the secretory function of goblet cells.

Proteomic-Level Responses of Early-Weaned Piglets to Dietary Arginine Supplementation

Dietary arginine supplementation enhances the immune status and protein synthesis in early-weaned pigs. However, the underlying mechanisms remain largely unknown. To investigate how arginine affects the expression of key proteins that regulate growth and nutrient transport of jejunum, a total of 12 healthy piglets （21 day-old, similar body weight, Landrace x Yorkshire） delivered by four sows were randomly divided into two groups. Piglets in the test group were reared with feed supplemented L-arginine at a concentration of 6.0 g/kg, while piglets in the control group were fed with feed supplemented L-alanine at a concentration of 12.3 g/kg （isonitrogenous control）. After 7 d, jejunum mucosae was collected and analyzed with the 2-D PAGE MS technology. Compared with the control pigs, arginine decreased the levels of proteins that regulate the protein syn- thesis, intermediary metabolism and tissue growth （ similar to anterior gradient 2 homolog, cyclophilin_ABHJike, hypothetical protein FLJ39502 and tetratrioopeptide repeat domain 16, similar to KIAA0156, mitechondrial ATP synthase, hydrion transporting F1 complex, beta subunit and alpha-tu- bulin ubiquitous isoform 19, prolyl 4-hydroxylase, beta subunit precursor）. In addition, arginine increased the levels of proteins that are involved in proteolysis and immune response （ PGAM1, T cell receptor beta variable 20, membrane steroid binding protein, similar to myomesin-1, and chain A, structure of pig muscle Pgk complexed with MgATP）. Therefore, arginine influences the immune response and protein synthesis mechanisms as well as improves eady weaned stress syndrome of piglets.

Composition,function,and timing:exploring the early‑life gut microbiota in piglets for probiotic interventions

Background The establishment of a robust gut microbiota in piglets during their early developmental stage holds the potential for long-term advantageous effects.However,the optimal timeframe for introducing probiotics to achieve this outcome remains uncertain.Results In the context of this investigation,we conducted a longitudinal assessment of the fecal microbiota of 63 piglets at three distinct pre-weaning time points.Simultaneously,we gathered vaginal and fecal samples from 23 sows.Employing 16S rRNA gene and metagenomic sequencing methodologies,we conducted a comprehensive analysis of the fluctuation patterns in microbial composition,functional capacity,interaction networks,and colonization resistance within the gut microbiota of piglets.As the piglets progressed in age,discernible modifications in intestinal microbial diversity,composition,and function were observed.A source-tracking analysis unveiled the pivotal role of fecal and vaginal microbiota derived from sows in populating the gut microbiota of neonatal piglets.By D21,the microbial interaction network displayed a more concise and efficient configuration,accompanied by enhanced colonization resistance relative to the other two time points.Moreover,we identified three strains of Ruminococcus sp.at D10 as potential candidates for improving piglets’weight gain during the weaning phase.Conclusions The findings of this study propose that D10 represents the most opportune juncture for the introduction of external probiotic interventions during the early stages of piglet development.This investigation augments our comprehension of the microbiota dynamics in early-life of piglets and offers valuable insights for guiding forthcoming probiotic interventions.

Effects of dietary Clostridium butyricum supplementation on growth performance,intestinal development,and immune response of weaned piglets challenged with lipopolysaccharide

Background: Weanling pigs, with immature immune system and physiological function, usually experience postweaning diarrhea. This study determined the effects of dietary Clostridium butyricum supplementation on growth performance, diarrhea, and immunity of weaned pigs challenged with lipopolysaccharide(LPS).Methods: In Experiment(Exp.) 1,144 weaned piglets were weaned at 21 d and randomly assigned to six groups,with six replicates per group and four pigs per replicate, receiving a control diet(CON) or diet supplemented with antibiotics(AB) or C. butyricum(CB)(0.1%, 0.2%, 0.4%, or 0.8%), respectively. All diets in Exp. 1 were a highly digestible basal diet, with 3,000 mg/kg zinc oxide supplied in the first 2 wk only. In Exp. 2, 180 piglets were weaned at 21 d and randomly assigned to five groups, with six replicates per group and six pigs per replicate, receiving CON, AB, or CB(0.2%, 0.4%, or 0.6%) diets. The digestibility of diets was lower than those in Exp. 1, and did not include zinc oxide. At 36 d of Exp. 2, 12 piglets were selected from each of the CON and 0.4% CB groups, six piglets were intraperitoneally injected with LPS(50 μg/kg body weight) and the other six piglets with normal saline;animals were killed at 4 h after injection to collect blood, intestine, and digesta samples for biochemical analysis.Results: In Exp. 1, CB and AB diets had no effect on growth performance of piglets. In Exp. 2, 0.4% CB decreased feed-gain ratio(P < 0.1), diarrhea score(P < 0.05), and increased duodenal, jejunal, and ileal villus height and jejunal villus height/crypt depth(P < 0.05). The 0.4% CB decreased the plasma tumor necrosis factor(TNF) α(P < 0.05) but increased ileal mucosa IL-10 and TLR2 mRNA expression(P < 0.05). Furthermore, 0.4% CB altered the microbial profile, with Bacillus and Ruminococcaceae UGG-003 at genus level and Lactobacillus casei and Parasutterella secunda at species level were higher than CON in colonic content(P < 0.05).Conclusions: Dietary C. butyricum supplementation had positive effects on growth of weaned piglets with less digestible diets. There was a tendency to reduce the feed-gain ratio, which could reduce feed costs in pig production. Moreover, C. butyricum decreased post-weaning diarrhea by improving the intestinal morphology,intestinal microflora profile, and immune function.

The effect of dietary tryptophan levels on oxidative stress of liver induced by diquat in weaned piglets

Oxidative stress can induce abnormal tryptophan metabolism. The present study was mainly conducted to determine the effect of dietary tryptophan levels on oxidative stress in the liver of weaned pigs challenged by diquat. A total of 36 PIC piglets weaned at 21 days of age were randomly allotted to 1 of 3 diets containing dietary tryptophan levels of 0.18, 0.30, and 0A5% for 14 d. On day 8, the piglets were injected intraperitoneally with sterile 0.9% NaCI solution or diquat （10 mg/kg body weight）. During the first 7 d of trial, increasing dietary tryptophan levels enhanced average daily gain （P = 0.09） and average daily feed intake （P = 0.08）, and decreased the feed efficiency （P 〈 0.05） of piglets. The growth performance was decreased by diquat injection （P 〈 0.05）. Diquat injection also decreased the activities of the superoxide dismutase （SOD） and glutathione peroxidase （GPx） in the plasma and liver （P 〈 0.05）, increased plasma malondialdehyde （MDA） （P 〈 0.05） and urea nitrogen （P 〈 0.05） concentrations, and enhanced MDA concentration （P = 0.09） and tryptophan 2,3-dioxygenase （TDO） activity （P = 0.07） in liver of piglets. Increasing dietary tryptophan levels could attenuate the effects of diquat injection on the MDA （P = 0.06） concentration and the activities of SOD （P = 0.09） and GPx （P = 0.05） of the liver, and plasma urea nitrogen （P = 0.06） concentration in the piglet. There was a synergistic role for increasing TDO activity in the liver between dietary tryptophan levels and diquat injection （P 〈 0.05）. These results suggest that increasing dietary tryptophan levels could attenuate the oxidative stress of the liver in weaned piglets intraperitoneally injected with diquat via enhancing the antioxidant capacity.

Effect of Dietary Supplementation with Hydrolyzed Wheat Gluten on Growth Performance, Cell Immunity and Serum Biochemical Indices of Weaned Piglets (Sus scrofa)

To investigate the effects of dietary supplementation with hydrolyzed wheat gluten （HWG） on growth performance, cell immunity and serum biochemical indices of weaned piglets, 160 crossed （Large White×andrace） and weaned piglets were randomly divided into 4 treatments with 4 replicates of 10 piglets each. The piglets in each treatment were fed an experimental diet containing either 0 g kg-1 HWG （control group）, 30 g kg-1 HWG （3% HWG group）, 50 g kg-1 HWG （5% HWG group）, or 2.5 g kg-1 glycyl-L-glutamine （0.25% Gly-Gln group）. The results showed that the diarrhea rates in 3% HWG and 5% HWG groups were significantly lower than in control group from d 1 to 14 （P〈0.05）, while the average daily gain （ADG） in each of two groups was increased （P〉0.05）. When compared with control group, dietary supplementation with 3% HWG increased the ratio of CD4＋：CD8＋ and the ratio of serum albumin and globulin concentrations （A：G） on d 14 and 28, as well as the proliferation of T- and B-lymphocytes （P〉0.05） on d 28. In addition, on d 14 and 28, the A：G ratio in 5% HWG group was significantly higher than in control group （P〈0.05）, while the ratio of CD4＋：CD8＋ increased slightly （P〉0.05）. Interestingly, 0.25% Gly-Gln group had higher proportion of CD3＋ （P〉0.05） and CD4＋ （P〈0.05） on d 14 than control group, but growth performances of 0.25% Gly-Gln group were negatively affected at all experiment stages. These results suggested that HWG might improve the growth performance of piglets by strengthening cell immunity and decreasing the occurrence of diarrhea during the prophase after weaning.

Effect of yeast Saccharomyces cerevisiae supplementation on serum antioxidant capacity, mucosal sIgA secretions and gut microbial populations in weaned piglets

This study was conducted to determine the effect of different forms of yeasts Saccharomyces cerevisiae supplementation on serum antioxidant capacity, mucosal secretory immunoglobulin A（s Ig A） secretions and gut microbial populations in weaned piglets. A total of 96 piglets weaned at 14 d of age were randomly allotted to 4 dietary treatments：（1） basal diet without yeast（Control）;（2） basal diet supplemented with 3.00 g kg–1 live yeast（LY）;（3） basal diet supplemented with 2.66 g kg–1 heat-killed whole yeast（HKY）; and（4） basal diet supplemented with 3.00 g kg–1 superfine yeast powders（SFY）. Each treatment had 4 replicates（pens）, with 6 piglets per replicate. The experiment lasted for 3 wk. At d 7 and 21 of the experiment, the samples of serum, mucosa and mesenteric lymph node（MLN） from jejunum, and digesta from the ileum and cecum were collected for determinations. Compared with the Control, dietary SFY supplementation increased serum superoxide dismutase（SOD） activity and lysozyme levels at d 7, and jejunum mucosal s Ig A secretions at d 21 of the experiment（P〈0.05）. Dietary LY supplementation increased serum SOD activity and jejunum mucosal s Ig A secretions, but decreased serum malondialdehyde（MDA） concentration at d 7 and 21（P〈0.05）. Piglets fed diets supplemented with LY and SFY had lower p H values and decreased numbers of Escherichia coli in the ileum and cecum contents at d 21 compared with the Control（P〈0.05）. Moreover, the ratio of Lactobacilli to E. coli in the ileum and cecum contents was increased by dietary LY and SFY supplementations（P〈0.05）. Collectively, different forms of yeasts, especially LY and SFY, may modulate body antioxidant capacity and enhance the intestinal immunity by regulation of secretions of mucosal s Ig A and reduction of pathogenic bacteria colonization, thus improving intestinal health of weaned piglets.

Administration of Saccharomyces boulardii mafic-1701 improves feed conversion ratio,promotes antioxidant capacity,alleviates intestinal inflammation and modulates gut microbiota in weaned piglets

Background:Probiotics are used as a means to improve animal health and intestinal development.Saccharomyces boulardii is a well-known probiotic;however,few studies have examined the effects of S.boulardii on weaned piglet performance.Therefore,this 28-day study compared the effects of S.boulardii mafic-1701 and aureomycin in diets for weaned piglets on growth performance,antioxidant parameters,inflammation and intestinal microbiota.One hundred and eight piglets,weaned at 28 d of age(8.5±1.1 kg),were randomly divided into the three dietary treatment groups with six pens and six piglets per pen(half male and half female).The dietary treatment groups were as follows:1)basal diet(CON);2)basal diet supplemented with 75 mg/kg aureomycin(ANT);3)basal diet supplemented with 1×108 CFU/kg S.boulardii mafic-1701(SB).Results:Compared to CON group,SB group had higher feed efficiency(P<0.05)in the last 14 d and lower diarrhea rate(P<0.05)over the entire 28 d.Total superoxide dismutase in serum was markedly increased in SB group(P<0.05).Moreover,compared with CON group,SB group decreased the levels of pro-inflammatory cytokines interleukin-6(P<0.01)and Tumor necrosis factor-α(P<0.05)in jejunum.Supplementation of S.boulardii mafic-1701 increased the abundance of Ruminococcaceae_UCG_009 and Turicibacter(P<0.05),whereas the abundance of unclassified_Clostridiaceae_4 was decreased(P<0.05).Furthermore,S.boulardii mafic-1701 administration increased cecal concentration of microbial metabolites,isobutyrate and valerate(P<0.05).Conclusions:The improvement in feed conversion ratio,reduction in diarrhea rate in weaned piglets provided diets supplemented with S.boulardii mafic-1701 may be associated with enhanced antioxidant activity,antiinflammatory responses and improved intestinal microbial ecology.

Tannic acid prevents post-weaning diarrhea by improving intestinal barrier integrity and function in weaned piglets

Background:Tannic acid(TA)is potential to reduce diarrhea in weaning pigs,but knowledge about the influence of TA on intestinal barrier integrity and function is still scarce.This experiment was conducted to investigate the effects of dietary TA supplementation on growth performance,diarrhea rate,intestinal barrier integrity and function of weaned pigs.Methods:A total of 108 crossbred(Duroc×Landrace×Yorkshire)piglets,with an initial average body weight of 6.60±0.27 kg,were allotted to 3 groups(6 pigs/pen and 6 replicates/group)in a randomized complete block design according to their gender and body weight.Piglets were fed the basal diet with 0(control,CON),0.2%and 1.0%TA,respectively.The trial lasted for 28 d.Results:Compared with the CON group,dietary 0.2%and 1.0%TA supplementation didn’t affect ADFI,ADG and F:G(P>0.05),but reduced diarrhea rate,diarrhea index and diarrhea score of piglets(P<0.05),reduced diamine oxidase(DAO)activity and D-lactic acid concentration in serum(P<0.01).The higher occludin expression and localization were observed in the duodenum,jejunum and ileum after supplementation with 0.2%or 1.0%TA(P<0.05).Adding 0.2%TA to diet significantly decreased crypt depth,increased villus height/crypt depth ratio in the duodenum(P<0.05),and dietary 1.0%TA tended to decrease crypt depth(P<0.10)and significantly decreased villus height(P<0.05)of the ileum.Moreover,lower malondialdehyde content in the ileum was detected in the pigs fed 1.0%TA(P<0.05).In the duodenum,both 0.2%and 1.0%TA groups had higher occludin(OCLN)mRNA and 0.2%TA group had higher zonula occludens-2(ZO-2)level(P<0.05).Meanwhile,dietary 1.0%TA supplementation tended to up-regulate OCLN mRNA levels in the jejunum(P<0.10)and 0.2%TA supplementation tended to up-regulate zonula occludens-1(ZO-1)mRNA levels in the ileum(P<0.10).Conclusion:In conclusion,dietary supplementation of 0.2%or 1.0%TA could effectively alleviate postweaning diarrhea without altering growth performance in weaned piglets,which might be achieved by improving intestinal barrier integrity and function.

Effects of Fermentation Compound Herbal Medicine on Production Performance and Immune Function of Weaned Piglets

The feed additive was composed of six Chinese Herbal（CH）（including Poria, Astragalus, Cork, Orange peel, Hawthorn and Radix）, which were fermented by Lactobacillus and used as a feed additive for piglets basal diet. Similar to group of antibiotic and herbal medicine, fermented herbal medicine was additived to basic diet in four trials with eight replications per trial, determining effect on the growth performance, biochemical and immunological indicators. Compared with the control group, the group of fermented herbal compound had higher Average Daily Gain（ADG）, higher content of immune globulin M（IgM） and immune globulin A（Ig A）（p〈0.05）. There was no significant effect on the indicators of ADG, Average Daily Feed Intake（ADFI） and diarrhea rate（p〉0.05） between groups of antibiotic and fermented herbal were observed; however, the concentrations of Total Protein（TP）, Growth Hormone（GH）, IgG, IgM and IgA significantly increased（p〈0.05） compared with the control group.

Tauroursodeoxycholic acid(TUDCA)improves intestinal barrier function associated with TGR5-MLCK pathway and the alteration of serum metabolites and gut bacteria in weaned piglets

Background:Tauroursodeoxycholic acid(TUDCA),a hydrophilic bile acid,is the main medicinal component of bear bile and is commonly used to treat a variety of hepatobiliary diseases.Meanwhile,TUDCA has been shown to modulate the intestinal barrier function and alleviate DSS-induced colitis in mice.However,the effect of TUDCA on the intestinal barrier of weaned piglets remains largely unclear.Methods:The weaned piglets and porcine IPEC-J2 intestinal epithelial cells were used to investigate the effects of TUDCA on intestinal barrier function in weaned piglets and explore the possible underlying mechanisms.In vivo,72 healthy weaned piglets were randomly allocated into 2 groups according to their gender and body weight,and piglets were fed the basal diet with 0(control,CON)and 200 mg/kg TUDCA for 30 d,respectively.Three female and three male piglets reflecting the average bodyweight were slaughtered in each group and samples were collected.In vitro,IPEC-J2 cells were subjected to 100μmol/L TUDCA to explore the possible underlying mechanisms.Results:Our results demonstrated that dietary TUDCA supplementation significantly reduced the diarrhea incidence of weaned piglets,possibly attributing to the TUDCA-enhanced intestinal barrier function and immunity.In addition,TUDCA supplementation altered serum metabolites and the relative abundance of certain gut bacteria,which might contribute to the improved intestinal barrier function.Furthermore,the in-vitro results showed that TUDCA improved the E.coli-induced epithelial barrier impairment of IPEC-J2 cells and increased Takeda G-coupled protein receptor 5(TGR5)protein expression.However,knockdown of TGR5 and inhibition of myosin light chain kinase(MLCK)pathway abolished the TUDCA-improved epithelial barrier impairment in E.coli-treated IPEC-J2 cells,indicating the involvement of TGR5-MLCK in this process.Conclusions:These findings showed that TUDCA improved intestinal barrier function associated with TGR5-MLCK pathway and the alteration of serum metabolites and gut bacteria in weaned piglets,suggesting the potential application of TUDCA in improving gut health in piglet production.

The prolonged effect of glucagon-like peptide 2 pretreatment on growth performance and intestinal development of weaned piglets

Background： Glucagon-like peptide 2 （GLP-2） is a potent epithelium-specific intestinal growth factor. The aim of this study was to demonstrate the prolonged effect of GLP-2 on the growth performance of weaned piglets. Forty piglets weaned at the age of 28 d with an average BW of 6.8 ＋ 0.4 kg were assigned to four treatments： （i） non- challenged control; （ii） LPS-challenged control; （iii） LPS ＋ low GLP-2; and （iv） LPS ＋ high GLP-2. Piglets in groups （i）, （ii）, and （iv） were s.c. injected with PBS supplemented with human [Gly2]GLP-21-34 at doses of 0, 2 and 10 nmol/kg BW per day for seven consecutive days. BW, gain：feed ratio （G：F）, and plasma GLP-2 levels were determined on d 0 7, and 14 after weaning. Piglets were challenged with i.p. administration of Escherichia coil lipopolysaccharide （LPS） at a dose of 100 pg/kg on d 14 to induce intestinal damage. Twenty-four hours later, intestinal tract samples were collected to assess intestinal morphology and quantify enzyme activity. Results： Plasma GLP-2 levels decreased after weaning, but in the high GLP-2 group, plasma GLP-2 was maintained on d 7 and even increased to a level higher than the preweaning level on d ]4 （P 〈 0.05）. High GLP-2 treatment significantly increased the duodenal, jejunal and ileal weight, as well as the gross weight of the small intestine （SI）, and the SI weight index （P 〈 0.05）. LPS caused villous atrophy and disrupted intestinal morphology in the duodenum, jejunum and ileum. GLP-2 also significantly increased the villus height and the villus height/crypt depth ratio （VCR） of the duodenum, jejunum, and ileum （P 〈 0.05）. Histological examination revealed that in GLP-2-treated groups, the integrity of the villus was maintained, and the villus was protected against LPS-induced damage. GLP-2 significantly increased the activity of alkaline phosphatase （AKP）, y-glutamyltranspeptidase （y-G-i-）, and pancreatic lipase in the duodenum and jejunum （P 〈 0.05）. GLP-2 treatment also significantly increased the average daily gain （ADG） and G：F of piglets at 0 to 7, 7 to 14, as well as 0 to14 d （P 〈 0.05）, resulting in a significant increase of final 8W in high GLP-2 pigs （P = 0.016）. Conclusions： Exogenous GLP-2 improved the growth of weaned piglets and protected them against LPS-induced intestinal damage. These effects may be due to the ability of GLP-2 to promote the secretion of endogenous GLP-2 to stimulate the small intestinal development.

Mechanisms Involved in the Growth Promotion of Weaned Piglets by High-level Zinc Oxide

Dietary supplementation with high lev- els of zinc oxide （ZnO） is an effective method to prevent diarrhea and promote growth in weaned pig- lets. However, the mechanisms behind these beneficial effects are not well understood. Recent advances in this field indicated that the effects of ZnO on piglet growth are achieved through multiple regulatory path- ways rather than a single pathway. This review focu- ses on recent advances in the effects of high level ZnO on： 1 ） Zn status in weaned piglets; 2 ） intestinal microbe flora regulation; 3 ） intestinal structure and function;4） ion secretion in the intestine epithelium;5） intestinal immunology; and 6） secretion of brain-in- testine peptides. ZnO appears to regulate the intestinal microflora and decreases the release of histamine to prevent diarrhea. ZnO also increases the expression of intestinal insulin-like growth factor-1 （ IGF-1 ） and the IGF-1 receptor to ameliorate intestinal injury associat- ed with weaning, and mediates secretion of brain-gut peptides to stimulate food intake, to promote the growth of piglets. Better understanding of the underly-1 ing mechanisms should inspire us to supplement the diet for weaned piglets with ZnO and investigate new approaches to prevent diarrhea and enhance growth.

Comparative effects of enzymatic soybean,fish meal and milk powder in diets on growth performance,immunological parameters,SCFAs production and gut microbiome of weaned piglets

Background:The objective of this study was to evaluate the replacement effects of milk powder(MK)and fish meal(FM)by enzymatic soybean(ESB)in diets on growth performance,immunological parameters,SCFAs production and gut microbiome of weaned piglets.Methods:A total of 128 piglets with initial body weight at 6.95±0.46 kg,were randomly assigned into 4 dietary treatments with 8 replicates per treatment and 4 piglets per replicate for a period of 14 d.Piglets were offered isonitrogenous and iso-energetic diets as follows:CON diet with MK and FM as high quality protein sources,ESB plus FM diet with ESB replacing MK,ESB plus MK diet with ESB replacing FM,and ESB diet with ESB replacing both MK and FM.Results:No significant differences were observed in growth performance among all treatments(P>0.05).However,piglets fed ESB plus FM or ESB diet had increased diarrhea index(P<0.01),and lower digestibility of dry matter(DM),gross energy(GE)or crude protein(CP),relative to piglets fed CON diet(P<0.01).Moreover,the inclusion of ESB in diet markedly decreased the plasma concentration of HPT and fecal concentration of butyric acid(BA)(P<0.01).The High-throughput sequencing of 16S rRNA gene V3−V4 region of gut microbiome revealed that the inclusion of ESB in diet increased the alpha diversity,and the linear discriminant analysis effect size(LEfSe)showed that piglets fed with ESB plus FM or ESB diet contained more gut pathogenic bacteria,such as g_Peptococcus,g_Veillonella and g_Helicobacter.Conclusion:The inclusion of ESB in diet did not markedly affect growth performance of piglets,but the replacement of MK or both MK and FM by ESB increased diarrhea index,which could be associated with lower nutrients digestibility and more gut pathogenic bacteria.However,piglets fed diet using ESB to replace FM did not markedly affect gut health-related parameters,indicating the potential for replacing FM with ESB in weaning diet.

Lactobacillus frumenti mediates energy production via fatty acid β-oxidation in the liver of early-weaned piglets

Background:Early-weaning of piglets is often accompanied by severe disorders,especially diarrhea.The gut microbiota and its metabolites play a critical role in the maintenance of the physiologic and metabolic homeostasis of the host.Our previous studies have demonstrated that oral administration of Lactobacillus frumenti improves epithelial barrier functions and confers diarrhea resistance in early-weaned piglets.However,the metabolic response to L.frumenti administration remains unclear.Then,we conducted simultaneous serum and hepatic metabolomic analyses in early-weaned piglets administered by L.frumenti or phosphatebuffered saline(PBS).Results:A total of 1006-day-old crossbred piglets(Landrace×Yorkshire)were randomly divided into two groups and piglets received PBS(sterile,2 m L)or L.frumenti(suspension in PBS,10~8 CFU/m L,2 m L)by oral administration once per day from 6 to 20 days of age.Piglets were weaned at 21 days of age.Serum and liver samples for metabolomic analyses were collected at 26 days of age.Principal components analysis(PCA)showed that L.frumenti altered metabolism in serum and liver.Numerous correlations(P<0.05)were identified among the serum and liver metabolites that were affected by L.frumenti.Concentrations of guanosine monophosphate(GMP),inosine monophosphate(IMP),and uric acid were higher in serum of L.frumenti administration piglets.Pathway analysis indicated that L.frumenti regulated fatty acid and amino acid metabolism in serum and liver.Concentrations of fatty acidβ-oxidation related metabolites in serum(such as3-hydroxybutyrylcarnitine,C4-OH)and liver(such as acetylcarnitine)were increased after L.frumenti administration.Conclusions:Our findings suggest that L.frumenti regulates lipid metabolism and amino acid metabolism in the liver of early-weaned piglets,where it promotes fatty acidβ-oxidation and energy production.High serum concentrations of nucleotide intermediates,which may be an alternative strategy to reduce the incidence of diarrhea in early-weaned piglets,were further detected.These findings broaden our understanding of the relationships between the gut microbiota and nutrient metabolism in the early-weaned piglets.

Effects of Supplemental Histamine on Gastric Acid Secretion, Digestive Enzyme Activities, Intestinal Microfloral of Early Weaned Piglets

Two experiments were conducted to study the effect of supplemental histamine in the diet of early-weaned piglets. In experiment A, 24 cross bred piglets with an average body weight of 6.10±0. 40 kg, weaned at the age of 28 days, were divided into four groups, fed with basal diet of low dietary copper without (control) or with supplemental histamine at 60, 120, 180μg kg-1 BW. During the two weeks and the third week after weaning, ADG(average daily gain) of piglets were increased by 15.8%(P<0. 05), 9.5%(P< 0.10) by addition of 60μg kg-1 BW histamine, but decreased by addition of 180μg kg-1BW histamine, which also increased the amount of E. coli in colon and the scour incidence. The secretion of gastric acid and pepsin were improved by both dose of supplemental histamine (60, 180μg kg-1BW) and gastric digesta pH were decreased by both. Addition of 60 μg kg-1 BW histamine improved the activities of trypsin, amylase in duodelum digesta. In experiment B, 12 cross bred piglets with an average body weight of 6. 85±0.35 kg, weaned at the age of 28 days, were divided into two groups, fed with basal diet of high dietary copper without (control) or with supplemental 60 μg kg-1 BW histamine. During the two weeks and the third week after weaning, ADG of piglets were increased by 9. 8% (P<0.05), 7. 0% (P<0. 10). The secretion of gastric acid, activities of trypsin and amylase in duodelum digesta, were also improved by addition of 60 μg kg-1BW histamine. The results showed that addition of histamine (60μg kg-1BW) in early weaned piglets could increase the secretion of gastric acid and pepsin, reduce gastric digesta pH and scour incidence, improve activities of trypsin, amylase in duodelum digesta, and the growing performance of early weaned piglets.

Estimation of endogenous protein and amino acid ileal losses in weaned piglets by regression analysis using diets with graded levels of casein

Background： Many studies have investigated endogenous loss of proteins and amino acids （AAs） at the ileal level in growing pigs. However, only a few studies have researched this subject in piglets. Knowledge regarding AA ileal digestibility in piglets would be helpful during the formulation of diets for weaning piglets, rather than just using coefficients obtained in growing pigs. Therefore, in this study, we sought to estimate endogenous protein and AA ileal losses in piglets. Furthermore, apparent and true ileal digestibility （AID and TID） of protein and AAs from casein were measured. Results： The average flow of protein was 20.8 g/kg of dry matter intake （DMI）. Basal protein loss, as estimated by regression, was 16.9 g/kg DMI. Glutamic acid, arginine, and aspartic acid （2.2, 1.4, and 1.2 g/kg DMI, respectively） were the AAs for which greater losses were seen. The AID of protein and AAs increased as the protein level in the diet increased. A higher increment in AID was observed between diets with 80 and160 g CP/kg of feed; this finding was mainly attributable to increases in glycine and arginine （46.1% and 18%, respectively）. The TID of protein was 97.8, and the TID of AAs varied from 93.9 for histidine to 100.2 for phenylalanine. Conclusions： The basal endogenous protein loss in piglets was 16.9 g/kg DMI. Endogenous protein was rich in glutamic acid, aspartic acid, and arginine, which represented 32.7% of endogenous protein loss in weaning piglets. The TID of casein was high and varied from 93.0 for histidine to 100.2 for phenylalanine.

The Absorption and Transportation of Glutamine Containing Small Peptides-Arg-Gly-GIn and Gly-GIn in Jejunum of Weaned Piglets

In order to research the absorption and transportation differences ofglutamine containing small peptides and free glutamine in intestine of weaned piglets, 28-day-old weaned piglets were selected to get the sample for jejunum culture in vitro in this study. Trial 1 was conducted to study the stability of tripeptide （Arg-Gly-Gln） in the jejunum to determine the ideal culture time. It was designed into 2 treatments： control group （Kreb＇s solution, GC） and tripeptide group （by adding 556μmol/L of Arg-Gly-Gln to Kreb＇s solution, GP）. They were cultured for 60 min. The absorption and transportation differences ofglutamine existing as Arg-Gly-Gln, Gly-Gln and free Gin were studied in trial 2. It was designed into 4 treatments： group 1 （G1, control group）, Kreb＇s solution; Group 2 （G2）, Kreb＇s solution＋556 μmol/L Arg-Gly-Gln; group 3 （G3）, Kreb＇s solution＋556 μmol/L Gly-Gln and Arg; group 4 （G4）, Kreb＇s solution＋556 μmol/L arginine, glycine and glutamine. They were cultured for 40 minutes. The results indicated that the absorption and transportation of glutamine existing as Arg-Gly-Gln was more efficient than those of Gly-Gln while Gly-Gln was more efficient than free glutamine. Arg-Gly-Gln may be more excellent than Gly-Gln in physiological functions and nutrition potential.