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.

Effects of different forms of yeast Saccharomyces cerevisiae on growth performance,intestinal development,and systemic immunity in early-weaned piglets

The present study was conducted to determine effects of different forms of yeast （Saccharomyces cerevisiae, strain Y200007） on the growth performance, intestinal development, and systemic immunity in early-weaned piglets. A total of 96 piglets （14-d old, initial average body weight of 4.5 kg） were assigned to 4 dietary treatments： （1） basa diet without yeast （Control）; （2） basal diet supplemented with 3.00 g/kg live yeast （LY）; （3） basal diet supplemented with 2.66 g/kg heat-killed whole yeast （HKY）; and （4） basal diet supplemented with 3.00 g/kg superfine yeast powders （SFY）. Diets and water were provided ad libitum to the piglets during 3-week experiment. Growth performance of piglets was measured weekly. Samples of blood and small intestine were collected at days 7 and 21 of experiment. Dietary supplementation with LY and SFY improved G：F of piglets at days ]-21 of the experiment （P 〈 0.05） compared to Control group. Serum concentrations of growth hormone （GH）, triiodothyronine （T3）, tetraiodothyronine （T4）, and insulin growth factor 1 （iGF-1） in piglets at day 21 of the experiment were higher when fed diets supplemented with LY and SFY than those in Control group （P 〈 0.05）. Compared to Control group, contents of serum urea nitrogen of piglets were reduced by the 3 yeast-supplemented diets （P 〈 0.05）. Diets supplemented with LY increased villus height and villus-to-crypt ratio in duodenum and jejunum of piglets （P 〈 0.05） compared to other two groups at day 7 of the experiment. Feeding diets supplemented with LY and SFY increased （P 〈 0.05） serum concentrations of IgA, IL-2, and IL-6 levels in piglets compared to Control. The CD4＋/CD8＋ ratio and proliferation of T-lymphocytes in piglets fed diets supplemented with LY were increased compared to that of Control group at day 7 of the experiment （P 〈 0.05）. In conclusion, dietary supplementation with both LY and SFY enhanced feed conversion, small intestinal development, and systemic immunity in early-weaned piglets, with better improvement in feed conversion by dietary supplementation with LY, while dietary supplementation with SFY was more effective in increasing systemic immune functions in early-weaned piglets.

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.

Effect of Glutamine Supplementation on Intestinal Mucosal Protein,DNA and Mucosal Morphology of Early-Weaned Piglets

Forty piglets weaned at 21 days age were randomly assigned to one of two groups. The treatment group contained 1. 2% glutamine. The diets were calculated to be isonitrogenous and isoenergetic, each treatment contained five replicates of four piglets. Five piglets from each dietary treatment were killed at 35 days and 49 days age. The results showed that dietary glutamine supplementation increased jejunal DNA content of piglets in the treatment group at 35 days and 49 days age. There was no significant change of ileum DNA content and intestinal protein content between the treatment and control ones. Dietary glutamine supplementation increased mucosal thickness of piglets in treatment group markedly at 35 days age. The intestinal villus heights were significantly higher in treatment than that in control. The results of electron micrograph showed that dietary glutamine supplementation prevented deterioration of intestinal epithelium, and maintained normal intestinal microvillus structure. The results suggested that glutamine supplementation prevented villus atrophy, and had protective effect on intestinal structure.

Replacing dietary sodium selenite with biogenic selenium nanoparticles improves the growth performance and gut health of early-weaned piglets

Selenium nanoparticles(SeNPs)are proposed as a safer and more effective selenium delivery system than sodium selenite(Na_(2)SeO_(3)).Here,we investigated the effects of replacing dietary Na_(2)SeO_(3)with SeNPs synthesized by Lactobacillus casei ATCC 393 on the growth performance and gut health of earlyweaned piglets.Seventy-two piglets(Duroc×Landrace×Large Yorkshire)weaned at 21 d of age were divided into the control group(basal diet containing 0.3 mg Se/kg from Na_(2)SeO_(3))and SeNPs group(basal diet containing 0.3 mg Se/kg from SeNPs)during a 14-d feeding period.The results revealed that SeNPs supplementation increased the average daily gain(P=0.022)and average daily feed intake(P=0.033),reduced(P=0.056)the diarrhea incidence,and improved(P=0.013)the feed conversion ratio compared with Na_(2)SeO_(3).Additionally,SeNPs increased jejunal microvilli height(P=0.006)and alleviated the intestinal barrier dysfunction by upregulating(P<0.05)the expression levels of mucin 2 and tight junction proteins,increasing(P<0.05)Se availability,and maintaining mitochondrial structure and function,thereby improving antioxidant capacity and immunity.Furthermore,metabolomics showed that SeNPs can regulate lipid metabolism and participate in the synthesis,secretion and action of parathyroid hormone,proximal tubule bicarbonate reclamation and tricarboxylic acid cycle.Moreover,SeNPs increased(P<0.05)the abundance of Holdemanella and the levels of acetate and propionate.Correlation analysis suggested that Holdemanella was closely associated with the regulatory effects of SeNPs on early-weaned piglets through participating in lipid metabolism.Overall,replacing dietary Na_(2)SeO_(3)with biogenic SeNPs could be a potential nutritional intervention strategy to prevent earlyweaning syndrome in piglets.

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.

The protective role of prebiotics and probiotics on diarrhea and gut damage in the rotavirus-infected piglets

Rotavirus is one of the pathogenic causes that induce diarrhea in young animals, especially piglets, worldwide. However, nowadays, there is no specific drug available to treat the disease, and the related vaccines have no obvious efficiency in some countries. Via analyzing the pathogenesis of rotavirus, it inducing diarrhea is mainly due to disturb enteric nervous system, destroy gut mucosal integrity, induce intracellular electrolyte imbalance, and impair gut microbiota and immunity. Many studies have already proved that prebiotics and probiotics can mitigate the damage and diarrhea induced by rotavirus infection in hosts. Based on these, the current review summarizes and discusses the effects and mechanisms of prebiotics and probiotics on rotavirus-induced diarrhea in piglets. This information will highlight the basis for the swine production utilization of prebiotics and probiotics in the prevention or treatment of rotavirus infection in the future.

Dietary supplementation of laminarin improves the reproductive performance of sows and the growth of suckling piglets

Background Maternal nutrition is essential in keeping a highly efficient production system in the pig industry.Laminarin has been shown to improve antioxidant capacity,reduce the inflammatory response,and favor the homeostasis of intestinal microbiota.However,the effect of dietary supplementation of laminarin on the reproductive performance of sows and the growth of suckling offspring remains unknown.Methods A total of 40 Landrace×Yorkshire multiparous sows on d 85 of gestation,similar in age,body weight(BW),parity and reproductive performance,were randomly divided into four dietary treatments with 10 sows per treatment,receiving a control diet(basal pregnancy or lactating diets)and a basal diet supplemented with 0.025%,0.05%and 0.10%laminarin,respectively.The experiment lasted from d 85 of gestation to d 21 of lactation.Results Laminarin supplementation linearly increased number born alive per litter(P=0.03),average daily feed intake(ADFI,P<0.01),and total milk yield of sows during the lactation of 1–21 d(P=0.02).Furthermore,maternal laminarin supplementation increased the average daily gain(ADG)of piglets while tending to reduce the culling and death rate before weaning.In addition,alterations to the composition of colostrum and milk,as well as to serum inflammatory cytokines and immunoglobulins of sows were observed.The fecal microbiota profile of sows supported the improvement of reproductive performance in sows and the growth performance in suckling offspring.Conclusions Dietary supplementation of laminarin during late pregnancy and lactation could significantly improve reproductive performance of sows and growth performance of piglets.

Effects of the kinetic pattern of dietary glucose release on nitrogen utilization, the portal amino acid profile, and nutrient transporter expression in intestinal enterocytes in piglets

Background Promoting the synchronization of glucose and amino acid release in the digestive tract of pigs could effectively improve dietary nitrogen utilization.The rational allocation of dietary starch sources and the exploration of appropriate dietary glucose release kinetics may promote the dynamic balance of dietary glucose and amino acid supplies.However,research on the effects of diets with different glucose release kinetic profiles on amino acid absorption and portal amino acid appearance in piglets is limited.This study aimed to investigate the effects of the kinetic pattern of dietary glucose release on nitrogen utilization,the portal amino acid profile,and nutrient transporter expression in intestinal enterocytes in piglets.Methods Sixty-four barrows(15.00±1.12 kg)were randomly allotted to 4 groups and fed diets formulated with starch from corn,corn/barley,corn/sorghum,or corn/cassava combinations(diets were coded A,B,C,or D respectively).Protein retention,the concentrations of portal amino acid and glucose,and the relative expression of amino acid and glucose transporter m RNAs were investigated.In vitro digestion was used to compare the dietary glucose release profiles.Results Four piglet diets with different glucose release kinetics were constructed by adjusting starch sources.The in vivo appearance dynamics of portal glucose were consistent with those of in vitro dietary glucose release kinetics.Total nitrogen excretion was reduced in the piglets in group B,while apparent nitrogen digestibility and nitrogen retention increased(P<0.05).Regardless of the time(2 h or 4 h after morning feeding),the portal total free amino acids content and contents of some individual amino acids(Thr,Glu,Gly,Ala,and Ile)of the piglets in group B were significantly higher than those in groups A,C,and D(P<0.05).Cluster analysis showed that different glucose release kinetic patterns resulted in different portal amino acid patterns in piglets,which decreased gradually with the extension of feeding time.The portal His/Phe,Pro/Glu,Leu/Val,Lys/Met,Tyr/Ile and Ala/Gly appeared higher similarity among the diet treatments.In the anterior jejunum,the glucose transporter SGLT1 was significantly positively correlated with the amino acid transporters B0AT1,EAAC1,and CAT1.Conclusions Rational allocation of starch resources could regulate dietary glucose release kinetics.In the present study,group B(corn/barley)diet exhibited a better glucose release kinetic pattern than the other groups,which could affect the portal amino acid contents and patterns by regulating the expression of amino acid transporters in the small intestine,thereby promoting nitrogen deposition in the body,and improving the utilization efficiency of dietary nitrogen.

Dietary supplementation of zinc oxide modulates intestinal functionality during the post-weaning period in clinically healthy piglets

Background To improve our understanding of host and intestinal microbiome interaction,this research investigated the effects of a high-level zinc oxide in the diet as model intervention on the intestinal microbiome and small intestinal functionality in clinically healthy post-weaning piglets.In study 1,piglets received either a high concentration of zinc(Zn)as zinc oxide(Zn O,Zn,2,690 mg/kg)or a low Zn concentration(100 mg/kg)in the diet during the post weaning period(d 14–23).The effects on the piglet's small intestinal microbiome and functionality of intestinal tissue were investigated.In study 2,the impact of timing of the dietary zinc intervention was investigated,i.e.,between d 0–14 and/or d 14–23 post weaning,and the consecutive effects on the piglet's intestinal functionality,here referring to microbiota composition and diversity and gene expression profiles.Results Differences in the small intestinal functionality were observed during the post weaning period between piglets receiving a diet with a low or high concentration Zn O content.A shift in the microbiota composition in the small intestine was observed that could be characterized as a non-pathological change,where mainly the commensals inter-changed.In the immediate post weaning period,i.e.,d 0–14,the highest number of differentially expressed genes(DEGs)in intestinal tissue were observed between animals receiving a diet with a low or high concentration Zn O content,i.e.,23 DEGs in jejunal tissue and 11 DEGs in ileal tissue.These genes are involved in biological processes related to immunity and inflammatory responses.For example,genes CD59 and REG3G were downregulated in the animals receiving a diet with a high concentration Zn O content compared to low Zn O content in both jejunum and ileum tissue.In the second study,a similar result was obtained regarding the expression of genes in intestinal tissue related to immune pathways when comparing piglets receiving a diet with a high concentration Zn O content compared to low Zn O content.Conclusions Supplementing a diet with a pharmaceutical level of Zn as Zn O for clinically healthy post weaning piglets influences various aspects intestinal functionality,in particular in the first two weeks post-weaning.The model intervention increased both the alpha diversity of the intestinal microbiome and the expression of a limited number of genes linked to the local immune system in intestinal tissue.The effects do not seem related to a direct antimicrobial effect of Zn O.

All-trans retinoic acid alleviates transmissible gastroenteritis virus-induced intestinal inflammation and barrier dysfunction in weaned piglets

Background Transmissible gastroenteritis virus(TGEV)is one of the main pathogens causing severe diarrhea of pig-lets.The pathogenesis of TGEV is closely related to intestinal inflammation.All-trans retinoic acid(ATRA)is the main active metabolite of vitamin A,which has immunomodulatory and anti-inflammatory properties.However,it is unclear whether ATRA can alleviate TGEV-induced intestinal inflammation and barrier dysfunction in piglets.This study aimed to investigate the effects of ATRA on growth performance,diarrhea,intestinal inflammation and intesti-nal barrier integrity of TGEV-challenged piglets.Methods In a 19-d study,32 weaned piglets were randomly divided into 4 treatments:Control group(basal diet),TGEV group(basal diet+TGEV challenge),TGEV+ATRA5 group(basal diet+5 mg/d ATRA+TGEV challenge)and TGEV+ATRA15 group(basal diet+15 mg/d ATRA+TGEV challenge).On d 14,piglets were orally administered TGEV or the sterile medium.Results Feeding piglets with 5 and 15 mg/d ATRA alleviated the growth inhibition and diarrhea induced by TGEV(P<0.05).Feeding piglets with 5 and 15 mg/d ATRA also inhibited the increase of serum diamine oxidase(DAO)activ-ity and the decrease of occludin and claudin-1 protein levels in jejunal mucosa induced by TGEV,and maintained intestinal barrier integrity(P<0.05).Meanwhile,5 mg/d ATRA feeding increased the sucrase activity and the expres-sions of nutrient transporter related genes(GLUT2 and SLC7A1)in jejunal mucosa of TGEV-challenged piglets(P<0.05).Furthermore,5 mg/d ATRA feeding attenuated TGEV-induced intestinal inflammatory response by inhibit-ing the release of interleukin(IL)-1β,IL-8 and tumor necrosis factor-α(TNF-α),and promoting the secretion of IL-10 and secretory immunoglobulin A(sIgA)(P<0.05).Feeding 5 mg/d ATRA also down-regulated the expressions of Toll-like receptors and RIG-I like receptors signaling pathway related genes(TLR3,TLR4,RIG-I,MyD88,TRIF and MAVS)and the phosphorylation level of nuclear factor-κB-p65(NF-κB p65),and up-regulated the inhibitor kappa B alpha(IκBα)protein level in jejunal mucosa of TGEV-challenged piglets(P<0.05).Conclusions ATRA alleviated TGEV-induced intestinal barrier damage by inhibiting inflammatory response,thus improving the growth performance and inhibiting diarrhea of piglets.The mechanism was associated with the inhibi-tion of NF-κB signaling pathway mediated by TLR3,TLR4 and RIG-I.

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.

Effect of Dietary Folic Acid Supplementation on Growth Performance and Hepatic Protein Metabolism in Early-Weaned Intrauterine Growth Retardation Piglets

To investigate the effects of dietary supplementation with folic acid on growth performance, hepatic protein metabolism and serum biochemical indices of early-weaned intrauterine growth retardation （IUGR） piglets, 24 male （Durocx （LandracexYorkshire）） weaned （14-d-old） IUGR piglets were randomly divided into 3 treatments with 8 replicates of 1 piglet per replicate. The piglets in each treatment were fed basal diet supplementation with either 0 （control）, 5 and 10 mg kg^-1 folic acid. The trial lasted for 21 d. Dietary folic acid supplementation reduced average daily feed intake （ADFI） （P〈0.05）. In addition, the average daily gain （ADG） in 10 mg kg^-1 folic acid group was significantly decreased （P〈0.01） and the ratio of feed：gain （F/G） increased slightly （P〉0.05）. Serum folic acid concentration increased （P〈0.01） with increasing folic acid inclusion, however, serum homocysteine concentration decreased significantly （P〈0.01）. Enhanced serum urine nitrogen （SUN） and diminished serum total protein （TP） as well as liver TP content were observed in 10 mg kg^-1 folic acid group （/＇〈0.05）. Furthermore, the relative mRNA expressions of insulin-like growth factor 1 （IGF-1） and mammalian target of rapamycin （m-TOR） in liver were respectively tended to reduce （P=0.06） and significantly downregulated （P〈0.05） in 10 mg kg1 group, in compared with 5 mg kg1 group. However, when compared with control group, folic acid supplementation had no significant effect on the mRNA abundance of IGF- 1 and m-TOR. The results indicated that supplementation with 10 mg kg-I folic acid impaired growth performance and hepatic protein metabolism of early-weaned IUGR piglets while 5 mg kg-~ folic acid enriched diet exerted limited positive effects.

Yeast hydrolysate attenuates lipopolysaccharide-induced inflammatory responses and intestinal barrier damage in weaned piglets

Background Intestinal inflammation is the main risk factor causing intestinal barrier dysfunction and lipopolysaccharide(LPS)can trigger inflammatory responses in various eukaryotic species.Yeast hydrolysate(YH)possesses multibiological effects and is received remarkable attention as a functional ingredient for improving growth performance and promoting health in animals.However,there is still inconclusive on the protective effects of dietary YH supplementation on intestinal barrier of piglets.This study was conducted to investigate the attenuate effects of YH supplementation on inflammatory responses and intestinal barrier injury in piglets challenged with LPS.Methods Twenty-four piglets(with an average body weight of 7.42±0.34 kg)weaned at 21 days of age were randomly assigned to one of two dietary treatments(12 replications with one pig per pen):a basal diet or a basal diet containing YH(5 g/kg).On the 22nd d,6 piglets in each treatment were intraperitoneally injected with LPS at 150μg/kg BW,and the others were injected with the same amount of sterile normal saline.Four hours later,blood samples of each piglet were collected and then piglets were euthanized.Results Dietary YH supplementation increased average daily feed intake and average daily gain(P<0.01),decreased the ratio of feed intake to gain of piglets(P sponse,evidenced by the increase o=0.048).Lipopolysaccharide(LPS)injection induced systemic inflammatory ref serum concentrations of haptoglobin(HP),adrenocorticotropic hormone(ACTH),cortisol,and interleukin-1β(IL-1β).Furthermore,LPS challenge resulted in inflammatory intestinal damage,by up-regulation of the protein or mRNA abundances of tumor necrosis factor-α(TNF-α),IL-1β,toll-like receptors 4(TLR4)and phosphor-nuclear factor-κB-p65(p-NFκB-p65)(P<0.01),and down-regulation of the jejunal villus height,the protein and mRNA abundances of zonula occludens-1(ZO-1)and occludin(OCC;P<0.05)in jejunal mucosa.Dietary YH supplementation decreased the impaired effects of ACTH,cortisol,HP,IL-1βand diamine oxidase in serum(P<0.05).Moreover,YH supplementation also up-regulated the jejunal villus height,protein and mRNA abundances of ZO-1 and OCC(P<0.05),down-regulated the mRNA expressions of TNF-αand IL-1βand the protein abundances of TNF-α,IL-1β,TLR4 and p-NFκB-p65 in jejunal mucosa in LPS-challenged pigs(P<0.01).Conclusion Yeast hydrolysate could attenuate inflammatory response and intestinal barrier injury in weaned piglets challenged with LPS,which was associated with the inhibition of TLR4/NF-κB signaling pathway activation.

Resveratrol alleviates oxidative stress induced by oxidized soybean oil and improves gut function via changing gut microbiota in weaned piglets

Background Oxidized soybean oil(OSO)has been shown to impair growth and exacerbate inflammation,leading to intestinal barrier injury in animals.Recent evidence suggests important roles for resveratrol(RES)in the promoting growth performance,antioxidant capacity,anti-inflammatory,and regulate intestinal barriers in animals.Therefore,The objectives of this study are to investigate the effects of dietary RES(purity 98%)supplementation on the growth performance,antioxidant capacity,inflammatory state,and intestinal function of weaned piglets challenged with OSO.Methods A total of 28 castrated weaned male piglets with a similar body weight of 10.197 replications per treatment and±0.10 kg were randomly assigned to 4 dietary treatments for 28-d feeding trial with 1 piglet per replicate.Treatments were arranged as a 2×2 factorial with oil type[3%fresh soybean oil(FSO)vs.3%OSO]and dietary RES(0vs.300 mg/kg).Results The results showed that relative to the FSO group,OSO stress tended to decrease the average daily feed intake(ADFI),and decreased the activity levels of lipase,villus/crypt ratio(VCR),the mRNA expression of FABP1,SOD2,IL-10 and ZO-1 in the jejunum,and SOD2,GPX1,occludin and ZO-1 in the colon,the levels of acetic acid in the colonic digesta,whereas up-regulated the mRNA expression of IL-1βand TNF-αin the jejunum(P<0.05).Moreover,dietary supplementation with RES increased ether extract(EE),the activity levels of sucrase,lipase,α-amylase,villus height(VH)and VCR,the mRNA expression of FABP1,SOD2,IL-10 and occludin in the jejunum,and FABP1,PPAR-γ,GPX1,occludin and ZO-1 in the colon,and the abundance of Firmicutes,acetic and propionic acid,but decreased the levels of D-lactic acid in the plasma,the abundance of Bacteroidetes in the colonic digesta of weaned piglets compared to the non-RES group(P<0.05).Meanwhile,in the interaction effect analysis,relative to the OSO group,dietary RES supplementation in the diets supplemented with OSO increased the activity levels of trypsin,VH in the jejunum,the abundance of Actinobacteria,the levels of butyric acid of weaned piglets,but failed to influence the activity levels of trypsin and VH,Actinobacteria abundance,the levels of butyric acid when diets were supplemented with FSO(interaction,P<0.05).Relative to the OSO group,dietary RES supplementation in the diets supplemented with OSO decreased the activity levels of DAO in the plasma of weaned piglets but failed to influence the activity levels of DAO when diets were supplemented with FSO(interaction,P<0.05).Relative to the FSO group,dietary RES supplementation in the diets supplemented with FSO decreased the level of propionic acid,whereas RES supplementation failed to influence the level of propionic acid when the diet was supplemented with OSO(interaction,P<0.01).Conclusions Inclusion of OSO intensified inflammatory states and impaired the intestinal health characteristics of weaned piglets.Dietary RES supplementation improved the antioxidant capacity,anti-inflammatory activity,and intestinal morphology.Further studies showed that the protective effects of RES on gut health could be linked to the decreased abundance of Prevotella_1,Clostridium_sensu_stricto_6,and Prevotellaceae_UCG003 and increased levels of acetic and propionic acid.

Supplemental Clostridium butyricum modulates lipid metabolism by reshaping the gut microbiota composition and bile acid profile in IUGR suckling piglets

Background Intrauterine growth restriction(IUGR)can cause lipid disorders in infants and have long-term adverse effects on their growth and development.Clostridium butyricum(C.butyricum),a kind of emerging probiotics,has been reported to effectively attenuate lipid metabolism dysfunctions.Therefore,the objective of this study was to investigate the effects of C.butyricum supplementation on hepatic lipid disorders in IUGR suckling piglets.Methods Sixteen IUGR and eight normal birth weight(NBW)neonatal male piglets were used in this study.From d 3to d 24,in addition to drinking milk,the eight NBW piglets(NBW-CON group,n=8)and eight IUGR piglets(IUGR-CON group,n=8)were given 10 mL sterile saline once a day,while the remaining IUGR piglets(IUGR-CB group,n=8)were orally administered C.butyricum at a dose of 2×108colony-forming units(CFU)/kg body weight(suspended in 10 mL sterile saline)at the same frequency.Results The IUGR-CON piglets exhibited restricted growth,impaired hepatic morphology,disordered lipid metabolism,increased abundance of opportunistic pathogens and altered ileum and liver bile acid(BA)profiles.However,C.butyricum supplementation reshaped the gut microbiota of the IUGR-CB piglets,characterized by a decreased abundance of opportunistic pathogens in the ileum,including Streptococcus and Enterococcus.The decrease in these bile salt hydrolase(BSH)-producing microbes increased the content of conjugated BAs,which could be transported to the liver and function as signaling molecules to activate liver X receptorα(LXRα)and farnesoid X receptor(FXR).This activation effectively accelerated the synthesis and oxidation of fatty acids and down-regulated the total cholesterol level by decreasing the synthesis and promoting the efflux of cholesterol.As a result,the growth performance and morphological structure of the liver improved in the IUGR piglets.Conclusion These results indicate that C.butyricum supplementation in IUGR suckling piglets could decrease the abundance of BSH-producing microbes(Streptococcus and Enterococcus).This decrease altered the ileum and liver BA profiles and consequently activated the expression of hepatic LXRαand FXR.The activation of these two signaling molecules could effectively normalize the lipid metabolism and improve the growth performance of IUGR suckling piglets.

The synbiotic mixture of lactulose and Bacillus coagulans protects intestinal barrier dysfunction and apoptosis in weaned piglets challenged with lipopolysaccharide

Background Lactulose as an effective prebiotic protects intestinal mucosal injury.Bacillus coagulans is widely used in feed additives because of its ability to promote intestinal health.Our previous study suggests that the combination of lactulose and Bacillus coagulans may be a good candidate as alternative for antibiotic growth promoters.However,the in vivo effects of lactulose and Bacillus coagulans on growth and intestinal health under immune challenge in piglets remains unclear.The objective of this study is to explore the protective effects of synbiotic containing lactu-lose and Bacillus coagulans on the intestinal mucosal injury and barrier dysfunction under immune challenge in weaned piglets.Methods Twenty four weaned piglets were assigned to 4 groups.Piglets in the CON-_(saline)and LPS-_(LPS)group were fed the basal diet,while others were fed either with chlortetracycline(CTC)or synbiotic mixture of lactulose and Bacillus coagulans for 32 d before injection of saline or lipopolysaccharide(LPS).Piglets were sacrificed 4 h after LPS injection to collect samples to determine intestinal morphology,integrity and barrier functions as well as relative genes and proteins.Results Our data showed that no differences were observed in the growth performance of the four test groups.LPS injection induced higher serum diamine oxidase activities,D-lactic acid levels,and endotoxin status,lower villus height and ratio of villus height to crypt depth,greater mRNA and lower protein expression related tight junction in both jejunum and ileum.In addition,a higher apoptosis index,and protein expression of Bax and caspase-3 were also observed in the LPS challenge group.Interestingly,dietary synbiotic mixture with lactulose and Bacillus coagulans protected against LPS-induced intestinal damage,barrier dysfunction and higher apoptosis as well as CTC.Conclusions Our data suggest that dietary supplementation of synbiotic mixture with lactulose and Bacillus coagu-lans showed resilience to LPS-induced intestinal morphological damage,barrier dysfunction and aggressive apoptosis in piglets as well as the protective effects of CTC.These results indicate that synbiotic mixture of lactulose and Bacillus coagulans showed beneficial effects on performance and resilience to acute immune stress in weaned piglets.

Dietary bile acid supplementation in weaned piglets with intrauterine growth retardation improves colonic microbiota,metabolic activity,and epithelial function

Background Intrauterine growth retardation(IUGR)is one of the major constraints in animal production.Our previ-ous study showed that piglets with IUGR are associated with abnormal bile acid(BA)metabolism.This study explored whether dietary BA supplementation could improve growth performance and colonic development,function,micro-biota,and metabolites in the normal birth weight(NBW)and IUGR piglets.A total of 48 weaned piglets(24 IUGR and 24 NBW)were allocated to four groups(12 piglets per group):(i)NBW group,(ii)NBW+BA group,(iii)IUGR group,and(iv)IUGR+BA group.Samples were collected after 28 days of feeding.Results The results showed that dietary BA supplementation increased the length and weight of the colon and colon weight to body weight ratio,while decreased the plasma diamine oxidase(DAO)concentration in the NBW pig-lets(P<0.05).Dietary BA supplementation to IUGR piglets decreased(P<0.05)the plasma concentrations of D-lactate and endotoxin and colonic DAO and endotoxin,suggesting a beneficial effect on epithelial integrity.Moreover,dietary BA supplementation to NBW and IUGR piglets increased Firmicutes abundance and decreased Bacteroidetes abundance(P<0.05),whereas Lactobacillus was the dominant genus in the colon.Metabolome analysis revealed 65 and 51 differential metabolites in the colon of piglets fed a diet with/without BA,respectively,which was associated with the colonic function of IUGR piglets.Furthermore,dietary BA supplementation to IUGR piglets upregulated the expressions of CAT,GPX,SOD,Nrf1,IL-2,and IFN-γin colonic mucosa(P<0.05).Conclusions Collectively,dietary BA supplementation could improve the colonic function of IUGR piglets,which was associated with increasing proportions of potentially beneficial bacteria and metabolites.Furthermore,BA shows a promising application prospect in improving the intestinal ecosystem and health of animals.

Supplementation with alpha-glycerol monolaurate during late gestation and lactation enhances sow performance,ameliorates milk composition,and improves growth of suckling piglets

Background Physiological changes during lactation cause oxidative stress in sows,reduce immunity,and hamper the growth capacity of piglets.Alpha-glycerol monolaurate(α-GML)has potential for enhancing the antimicrobial activity of sows and the growth of suckling piglets.Methods Eighty sows were allocated randomly to four groups:basal diet and basal diets supplemented with 500,1000,or 2000 mg/kgα-GML.The experiment started on d 85 of gestation and lasted until piglets were weaned on d 21 of lactation.The number of live-born piglets was standardized to 12 tion,body weight of piglets was measured and milk samples±1 per sow on day of parturition.On d 0 and21 of lactawere obtained from sows,and serum samples and feces from piglets were obtained on d 21.Results Feed intake,backfat loss,and weaning estrus interval did not differ among the four groups of sows.Maternalα-GML supplementation increased(P<0.05)the body weight of piglets at weaning and the apparent total tract digestibility of crude fat of sows.The immunoglobulin A and immunoglobulin G levels were greater(P<0.05)in a quadratic manner in the milk of sows as dietaryα-GML increased.Concerning fatty acid profile,C12:0,C15:0,C17:0,C18:2n6c,C18:3n3,C24:0,and C22:6n3 were higher(P<0.05)in linear and quadratic manners in colostrum of sowsfedα-GML diets compared with the control sows.There was lower(P<0.05)n-6:n-3 polyunsaturated fatty acid ratio in milk than in the control sows.Maternalα-GML increased the abundance of Firmicutes(P<0.05)and decreased the abundance of Proteobacteria(P<0.05)of piglet fecal microbiota.Conclusions Dietary supplementation withα-GML improved milk immunoglobulins and altered fatty acids of sows,thereby improving the health of piglets.

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.