Subpopulations of regulatory T cells are associated with subclinical atherosclerotic plaques,levels of LDL,and cardiorespiratory fitness in the elderly

Background:Atherosclerosis forms the pathological basis for the development of cardiovascular disease.Since pathological processes initially develop without clinically relevant symptoms,the identification of early markers in the subclinical stage plays an important role for initiating early interventions.There is evidence that regulatory T cells(Tregs)are involved in the development of atherosclerosis.Therefore,the present study aimed to identify and investigate associations with Tregs and their subsets in a cohort of healthy elderly individuals with and without subclinical atherosclerotic plaques(SAP).In addition,various lifestyle and risk factors,such as cardiorespiratory fitness,were investigated as associated signatures.Methods:A cross-sectional study was performed in 79 participants(male:n=50;age=63.6±3.7 years;body mass index=24.9±3.1 kg/m2;mean±SD)who had no previous diagnosis of chronic disease and were not taking medication.Ultrasound of the carotids to identify SAP,cardiovascular function measurement for vascular assessment and a cardiorespiratory fitness test to determine peak oxygen uptake were performed.Additionally,tests were conducted to assess blood lipids and determine glucose levels.Immunophenotyping of Tregs and their subtypes(resting(rTregs)and effector/memory(mTregs))was performed by 8-chanel flow cytometry.Participants were categorized according to atherosclerotic plaque status.Linear and logistic regression models were used to analyze associations between parameters.Results:SAP was detected in a total of 29 participants.The participants with plaque were older(64.8±3.6 years vs.62.9±3.5 years)and had higher peripheral systolic blood pressure(133.8±14.7 mmHg vs.125.8±10.9 mmHg).The participants with SAP were characterized by a lower percentage of rTregs(28.8%±10.7%vs.34.6%±10.7%)and a higher percentage of mTregs(40.3%±14.7%vs.30.0%±11.9%).Multiple logistic regression identified age(odds ratio(OR)=1.20(95%confidence interval(95%CI):1.011.42))and mTregs(OR=1.05(95%CI:1.021.10))as independent risk factors for SAP.Stepwise linear regression could reveal an association of peak oxygen uptake(β=0.441),low-density lipoprotein(LDL)(β=0.096),and SAP(β=6.733)with mTregs and LDL(β=0.104)with rTregs.Conclusion:While at an early stage of SAP,the total proportion of Tregs gives no indication of vascular changes,this is indicated by a shift in the Treg subgroups.Factors such as serum LDL or cardiopulmonary fitness may be associated with this shift and may also be additional diagnostic indicators.This could be used to initiate lifestyle-based preventive measures at an early stage,which may have a protective effect against disease progression.

Standardized ileal digestibility of amino acids in soybean meal fed to non-pregnant and pregnant sows

Background Two studies were designed to determine standard ileal crude protein(CP)and amino acid(AA)digestibility of soybean meal(SBM)from different origins fed to non-pregnant and pregnant sows.Seven solvent-extracted SBMs from soybeans produced in the USA,Brazil,and China were selected.In Exp.1,eight different diets were created:a nitrogen(N)-free diet and 7 experimental diets containing SBM from different origins as the only N source.Eight non-pregnant,multiparous sows were arranged in an 8×8 Latin square design(8 periods and 8 diets).In Exp.2,the diet formula was the same as in Exp.1.Eight gestating sows(parity 3)were assigned to 4 different diets in a replicated 4×3 Youden square design(three periods and four diets)in mid-gestation and again in late-gestation stages.Results When fed to non-pregnant and late-gestating sows,the standardized ileal digestibility(SID)of CP and most AAs from different SBM were not significantly different(P>0.05).When fed to mid-gestating sows,the SID values for Arg,His,Lys,Phe,Cys,Gly,Ser,and Tyr in SBM 1 were lower than in SBM 4 and 5(P<0.05),whereas SID for Leu from SBM 5 was higher than in SBM 1 and 4(P<0.05).SID values for Ile,Ala,and Asp from SBM 4 were lower than in SBM 1 and 5(P<0.05).Sows had significantly greater SID values for Lys,Ala,and Asp during mid-gestation when compared with late-gestation stages(P<0.05).Mid-gestating sows had greater SID value for Val and lower SID value for Tyr when compared with non-pregnant and late-gestating sows(P<0.01),whereas non-pregnant sows had significantly greater SID value for Met when compared with gestating sows(P<0.01).Conclusions When fed to mid-gestating sows,the SID values for most AAs varied among SBM samples.The SID values for Lys,Met,Val,Ala,Asp,and Tyr in SBM were affected by sow gestation stages.Our findings provide a cornerstone for accurate SBM use in sow diets.

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.

Effects of Weaning on Immunity and Nutrition in Piglets

The number of birth per year was an important indicator of the sow＇s productivity and it was related to the duration of breasffeeding. Weaning had been shown to be a stressful period. The weaning age had declined over time in order to increase sow productivity and the fact that weaning under modern-day commercial conditions inflicted stress （environmental, nutritional and psychological/social） on pigs and associated with marked changes in gastrointestinal tract （GIT） physiology, microbiology and immunology. However, the early weaning resulted in post-weaning diarrhea, mainly related to intestinal dysbiosis during this rapid adaptation to the new diet. The environment and feed had influence on physiology, gut morphology, immune system and diseases of weaned piglets.

Dietary supplementation of Macleaya cordata extract and Bacillus in combination improve laying performance by regulating reproductive hormones,intestinal microbiota and barrier function of laying hens

Background:This study aimed to investigate whether the combination of Macleaya cordata extract(MCE)and Bacil-lus could improve the laying performance and health of laying hens better.Methods:A total of 36029-week-old Jingbai laying hens were randomly divided into 4 treatments:control group(basal diet),MCE group(basal diet+MCE),Probiotics Bacillus Compound(PBC)group(basal diet+compound Bacil-lus),MCE+PBC group(basal diet+MCE+compound Bacillus).The feeding experiment lasted for 42 d.Results:The results showed that the laying rate and the average daily egg mass in the MCE+PBC group were significantly higher than those in the control group(P<0.05)and better than the MCE and PBC group.Combina-tion of MCE and Bacillus significantly increased the content of follicle-stimulating hormone(FSH)in the serum and up-regulated the expression of related hormone receptor gene(estrogen receptor-β,FSHR and luteinizing hormone/choriogonadotropin receptor)in the ovary of laying hens(P<0.05).In the MCE+PBC group,the mRNA expressions of zonula occluden-1,Occludin and mucin-2 in jejunum was increased and the intestinal epithelial barrier detected by transmission electron microscopy was enhanced compared with the control group(P<0.05).In addition,compared with the control group,combination of MCE and Bacillus significantly increased the total antioxidant capacity and catalase activity(P<0.05),and down-regulated the mRNA expressions of inflammation-related genes(interleukin-1βand tumor necrosis factor-α)as well as apoptosis-related genes(Caspase 3,Caspase 8 and P53)(P<0.05).The concen-tration of acetic acid and butyric acid in the cecum content of laying hens in the MCE+PBC group was significantly increased compared with the control group(P<0.05).Conclusions:Collectively,dietary supplementation of 600μg/kg MCE and 5×108 CFU/kg compound Bacillus can improve laying performance by improving microbiota to enhance antioxidant capacity and intestinal barrier,regulate reproductive hormones and the concentration of cecal short-chain fatty acids of laying hens,and the combined effect of MCE and Bacillus is better than that of single supplementation.

Effects of different starch structures on energy metabolism in pigs

Background Starch is a major component of carbohydrates and a major energy source for monogastric animals.Starch is composed of amylose and amylopectin and has different physiological functions due to its different structure.It has been shown that the energy supply efficiency of amylose is lower than that of amylopectin.However,there are few studies on the effect of starch structure on the available energy of pigs.The purpose of this study was to measure the effect of different structures of starch in the diet on the net energy(NE)of pigs using a comparative slaughter method and to establish a prediction equation to estimate the NE of starch with different structures.Fifty-six barrows(initial BW 10.18±0.11 kg)were used,and they were housed and fed individually.Pigs were divided into 7 treatments,with 8 replicates for each treatment and 1 pig for each replicate.One of the treatments was randomly selected as the initial slaughter group(ISG).Pigs in the remaining treatments were assigned to 6 diets,fed with basic diet and semi-pure diets with amylose/amylopectin ratio(AR)of 3.09,1.47,0.25,0.15 and 0.12,respectively.The experiment lasted for 28 d.Results Results showed that compared with the high amylose(AM)groups(AR 3.09 and 1.47),the high amylopectin(AP)group(AR 0.15)significantly increased the final BW,average daily weight gain and average daily feed intake of pigs(P<0.05),but the F:G of the AM group was lower(P<0.01).In addition,AR 0.15 and 0.12 groups have higher(P<0.01)nutrient digestibility of dry matter,crude protein,gross energy and crude ash.Meanwhile,compared with other groups,AR 0.15 group has a higher(P<0.05)NE intake and energy retention(RE).The regressive equation for predicting with starch structures was established as RE=1,235.243-48.298AM/AP(R^(2)=0.657,P=0.05).Conclusions In conclusion,NE intake and RE of pigs augmented with the increase of dietary amylopectin content,indicating that diets high in amylopectin were more conducive to promoting the growth of pigs in the late conserva-tion period.

Exopolysaccharides of Lactobacillus rhamnosus GG ameliorate Salmonella typhimurium-induced intestinal inflammation via the TLR4/NF-κB/MAPK pathway

Background Salmonella typhimurium(S.T),as an important foodborne bacterial pathogen,can cause diarrhea and gastroenteritis in humans and animals.Numerous studies have confirmed that exopolysaccharides(EPSs)have various biological functions,but the mechanism through which EPSs improve the immunity of animals against the invasion of pathogenic bacteria is unclear.Here,we explored the protective effect of EPSs of Lactobacillus rhamnosus GG(LGG)on the S.T-infected intestine.Methods Mice received adequate food and drinking water for one week before the start of the experiment.After 7 d of prefeeding,2×108 CFU/mL S.T solution and an equivalent volume of saline(control group)were given orally for 1 d.On the fourth day,the mice were treated with 0.5 mg/mL EPSs,1.0 mg/mL EPSs,2.0 mg/mL EPSs,or 2.0 mg/mL penicillin for 7 d.Finally,the body and relative organ weight,histological staining,and the levels of antioxidant enzyme activity and inflammatory cytokines were determined.Results The S.T-infected mice exhibited symptoms of decreased appetite,somnolence,diarrhea and flagging spirit.Treatment with EPSs and penicillin improved the weight loss of the mice,and the high dose of EPSs showed the best therapeutic effect.EPSs significantly ameliorated S.T-induced ileal injury in mice.High-dose EPSs were more effective than penicillin for alleviating ileal oxidative damage induced by S.T.The mRNA levels of inflammatory cytokines in the ileum of mice showed that the regulatory effects of EPSs on inflammatory cytokines were better than those of penicillin.EPSs could inhibit the expression and activation of key proteins of the TLR4/NF-κB/MAPK pathway and thereby suppress the level of S.T-induced ileal inflammation.Conclusions EPSs attenuate S.T-induced immune responses by inhibiting the expression of key proteins in the TLR4/NF-κB/MAPK signaling pathway.Moreover,EPSs could promote bacterial aggregation into clusters,which may be a potential strategy for reducing the bacterial invasion of intestinal epithelial cells.

Effect of replacement of soybean oil by Hermetia illucens fat on performance,digestibility,cecal microbiome,liver transcriptome and liver and plasma lipidomes of broilers

Background In contrast to protein-rich insect meal,the feed potential of insect fat is generally less explored and knowledge about the suitability of insect fat as a fat source specifically in broiler diets is still limited.In view of this,the present study aimed to comprehensively investigate the effect of partial(50%) and complete replacement of soybean oil with insect fat from Hermetia illucens(HI) larvae in broiler diets on performance,fat digestibility,cecal microbiome,liver transcriptome and liver and plasma lipidomes.Thus,100 male,1-day-old Cobb 500 broilers were randomly assigned to three groups and fed three different diets with either 0(group HI-0,n cens(HI) larvae fat for 35 d.= 30),2.5%(group HI-2.5,n %(HI-5.0,n = 35) or 5.0= 35) Hermetia illuResults Body weight gain,final body weight,feed intake,and feed:gain ratio during the whole period and apparent ileal digestibility coefficient for ether extract were not different between groups.Cecal microbial diversity did not differ between groups and taxonomic analysis revealed differences in the abundance of only four low-abundance bacterial taxa among groups;the abundances of phylum Actinobacteriota,class Coriobacteriia,order Coriobacteriales and family Eggerthellaceae were lower in group HI-5.0 compared to group HI-2.5(P < 0.05).Concentrations of total and individual short-chain fatty acids in the cecal digesta were not different between the three groups.Liver transcriptomics revealed a total of 55 and 25 transcripts to be differentially expressed between groups HI-5.0 vs.HI-0 and groups HI-2.5 vs.HI-0,respectively(P < 0.05).The concentrations of most lipid classes,with the exception of phosphatidylethanolamine,phosphatidylglycerol and lysophosphatidylcholine in the liver and cholesterylester and ceramide in plasma(P < 0.05),and of the sum of all lipid classes were not different between groups.Conclusions Partial and complete replacement of soybean oil with HI larvae fat in broiler diets had no effect on growth performance and only modest,but no adverse effects on the cecal microbiome and the metabolic health of broilers.This suggests that HI larvae fat can be used as an alternative fat source in broiler diets,thereby,making broiler production more sustainable.

Suitability of Hermetia illucens larvae meal and fat in broiler diets:effects on animal performance,apparent ileal digestibility,gut histology,and microbial metabolites

Background:The possibility of partially replacing soybean meal(SBM)with Hermetia illucens(HI)defatted larvae meal in broiler nutrition has frequently been suggested.For sustainability reasons,however,the larvae fat produced during defatting should also be used and could be particularly beneficial regarding gut health due to its fatty acid composition.To evaluate the suitability of HI larvae as protein and fat source,a 2×3 factorial arrangement with two types of protein,i.e.SBM(S)or SBM and 15%of its crude protein replaced by HI larvae meal(L),and three levels of fat sources,namely 0(0 L),50%(50 L)or 100%HI larvae fat(100 L)at the expense of soybean oil was applied.Results:In the starter phase,an interaction showed higher body weight(BW),average daily gain(ADG)and improved feed conversion ratio(FCR)if 50%or 100%HI larvae fat was fed with HI larvae meal.Moreover,BW,ADG and FCR improved when feeding HI larvae meal as protein source.Additionally,we observed an increased average daily feed intake in the grower,finisher,and overall phase in the L groups and an improved FCR in 0 L compared to 50 L groups during the overall period.Regarding apparent ileal digestibility,HI larvae meal feeding increased dry matter,organic matter,and fat digestibility.Feeding HI larvae meal as protein source decreased the concentrations of agmatine,spermidine,spermine and ammonia in the caecal digesta,whereas fat source affected agmatine with higher concentrations in 50 L compared to 0 L in the colonic digesta.In contrast,caecal ethanolamine concentrations increased in HI larvae meal groups compared to SBM.Caecal butyric acid concentrations decreased with HI larvae meal feeding.An interaction was found for the jejunal villus area,being higher in L+100 L compared to S+100 L.Furthermore,L groups had greater villus width.Conclusions:A partial replacement of SBM with HI larvae meal and soybean oil with HI larvae fat in broiler diets without impairing animal performance or gut health seems possible.Feeding HI larvae meal affected broiler performance positively in the starter phase and improved apparent ileal digestibility.

Heat stress in pigs and broilers:role of gut dysbiosis in the impairment of the gut-liver axis and restoration of these effects by probiotics,prebiotics and synbiotics

Heat stress is one of the most challenging stressors for animal production due to high economic losses resulting from impaired animal’s productivity,health and welfare.Despite the fact that all farm animal species are susceptible to heat stress,birds and pigs are particularly sensitive to heat stress due to either lacking or non-functional sweat glands.Con-vincing evidence in the literature exists that gut dysbiosis,a term used to describe a perturbation of commensal gut microbiota,develops in broilers and pigs under heat stress.Owing to the protective role of commensal bacteria for the gut barrier,gut dysbiosis causes a disruption of the gut barrier leading to endotoxemia,which contributes to the typical characteristics of heat stressed broilers and growing and growing-finishing pigs,such as reduced feed intake,decreased growth and reduced lean carcass weight.A substantial number of studies have shown that feeding of probiotics,prebiotics and synbiotics is an efficacious strategy to protect broilers from heat stress-induced gut barrier disruption through altering the gut microbiota and promoting all decisive structural,biochemical,and immunologi-cal elements of the intestinal barrier.In most of the available studies in heat stressed broilers,the alterations of gut microbiota and improvements of gut barrier function induced by feeding of either probiotics,prebiotics or synbiot-ics were accompanied by an improved productivity,health and/or welfare when compared to non-supplemented broilers exposed to heat stress.These findings indicate that the restoration of gut homeostasis and function is a key target for dietary interventions aiming to provide at least partial protection of broilers from the detrimental impact of heat stress conditions.Despite the fact that the number of studies dealing with the same feeding strategy in heat stressed pigs is limited,the available few studies suggest that feeding of probiotics might also be a suitable approach to enhance productivity,health and welfare in pigs kept under heat stress conditions.

Dietary fiber during gestation improves lactational feed intake of sows by modulating gut microbiota

Background The feed intake of sows during lactation is often lower than their needs.High-fiber feed is usually used during gestation to increase the voluntary feed intake of sows during lactation.However,the mechanism underlying the effect of bulky diets on the appetites of sows during lactation have not been fully clarified.The current study was conducted to determine whether a high-fiber diet during gestation improves lactational feed intake(LFI)of sows by modulating gut microbiota.Methods We selected an appropriate high-fiber diet during gestation and utilized the fecal microbial transplanta-tion(FMT)method to conduct research on the role of the gut microbiota in feed intake regulation of sows during lactation,as follows:high-fiber(HF)diet during gestation(n=23),low-fiber(LF)diet during gestation(n=23),and low-fiber diet+HF-FMT(LFM)during gestation(n=23).Results Compared with the LF,sows in the HF and LFM groups had a higher LFI,while the sows also had higher peptide tyrosine tyrosine and glucagon-like peptide 1 on d 110 of gestation(G110 d).The litter weight gain of piglets during lactation and weaning weight of piglets from LFM group were higher than LF group.Sows given a HF diet had lower Proteobacteria,especially Escherichia-Shigella,on G110 d and higher Lactobacillus,especially Lactobacil-lus_mucosae_LM1 and Lactobacillus_amylovorus,on d 7 of lactation(L7 d).The abundance of Escherichia-Shigella was reduced by HF-FMT in numerically compared with the LF.In addition,HF and HF-FMT both decreased the perinatal concentrations of proinflammatory factors,such as endotoxin(ET),lipocalin-2(LCN-2),tumor necrosis factor-α(TNF-α),and interleukin-1β(IL-1β).The concentration of ET and LCN-2 and the abundance of Proteobacteria and Escheri-chia-Shigella were negatively correlated with the LFI of sows.Conclusion The high abundance of Proteobacteria,especially Escherichia-Shigella of LF sows in late gestation,led to increased endotoxin levels,which result in inflammatory responses and adverse effects on the LFI of sows.Add-ing HF during gestation reverses this process by increasing the abundance of Lactobacillus,especially Lactobacil-lus_mucosae_LM1 and Lactobacillus_amylovorus.

Winery by‑products as a feed source with functional properties:dose–response effect of grape pomace,grape seed meal,and grape seed extract on rumen microbial community and their fermentation activity in RUSITEC

Background Grape and winery by-products have nutritional values for cattle and also contain functional compounds like phenols,which not only bind to protein but can also directly affect microbiota and their function in the rumen.We characterized the nutritional and functional effects of grape seed meal and grape pomace as well as an effective dosage of grape phenols on ruminal microbiota and fermentation characteristics using a rumen simulation technique.Results Six diets(each n=8)were compared including a control diet(CON,no by-product),a positive control diet(EXT,CON+3.7%grape seed extract on a dry matter(DM)basis),two diets with grape seed meal at 5%(GS-low)and 10%(GS-high),and two diets with grape pomace:at 10%(GP-low)and 20%(GP-high),on a DM basis.The inclusion of the by-product supplied total phenols at 3.4%,0.7%,1.4%,1.3%,and 2.7%of diet DM for EXT,GS-low,GS-high,GP-low,and GP-high,respectively.Diets were tested in four experimental runs.All treatments decreased ammonia concentrations and the disappearances of DM and OM(P<0.05)compared to CON.EXT and GP-high lowered butyrate and odd-and branch-chain short-chain fatty acids while increased acetate compared to CON(P<0.05).Treatments did not affect methane formation.EXT decreased the abundance of many bacterial genera including those belonging to the core microbiota.GP-high and EXT consistently decreased Olsenella and Anaerotipes while increased Ruminobacter abundances.Conclusion The data suggest that the inclusion of winery by-products or grape seed extract could be an option for reducing excessive ammonia production.Exposure to grape phenols at a high dosage in an extract form can alter the rumen microbial community.This,however,does not necessarily alter the effect of grape phenols on the microbial community function compared to feeding high levels of winery by-products.This suggests the dominant role of dosage over the form or source of the grape phenols in affecting ruminal microbial activity.In conclusion,supplementing grape phenols at about 3%of diet DM is an effective dosage tolerable to ruminal microbiota.

Changes of the liver metabolome following an intravenous lipopolysaccharide injection in Holstein cows supplemented with dietary carnitine

Background:Carnitine facilitates the flux of long-chain fatty acids for hepatic mitochondrial beta-oxidation,which acts to ameliorate the negative energy balance commonly affecting high-yielding dairy cows.Inflammation triggered by lipopolysaccharide(LPS)load can however pose a challenge to the metabolic integrity via the expression of pro-inflammatory mediators,leading to immune system activation and respective metabolic alterations.The effect of enhanced carnitine availability on hepatic metabolome profiles during an inflammatory challenge has not yet been determined in dairy cows.Herein,Holstein cows were supplemented with 25 g/d rumen-protected carnitine from 42d prepartum until 126 d postpartum(n=16)or assigned to the control group with no supplementation during the same period(n=14).We biopsied the liver of the cows before(100 d postpartum)and after(112 d postpartum)an intravenous injection of 0.5μg/kg LPS.Liver samples were subjected to a targeted metabolomics analysis using the AbsoluteIDQ p180 Kit(Biocrates Life Sciences AG,Innsbruck,Austria).Results:Multivariate statistical analyses revealed that hepatic metabolome profiles changed in relation to both the carnitine supplementation and the LPS challenge.Comparing the metabolite profiles on 100 d,carnitine increased the concentration of short-and long-chain acyl-carnitines,which may be explained by an enhanced mitochondrial fatty acid shuttle and hence greater energy availability.The LPS injection affected hepatic metabolite profiles only in the carnitine supplemented group,particularly altering the concentration of biogenic amines.Conclusions:Our results point to interactions between an acute hepatic inflammatory response and biogenic amine metabolism,depending on energy availability.

Zinc glycine chelate ameliorates DSS‑induced intestinal barrier dysfunction via attenuating TLR4/NF‑κB pathway in meat ducks

Background Zinc glycine chelate(Zn-Gly)has anti-inflammation and growth-promoting properties;however,the mechanism of Zn-Gly contribution to gut barrier function in Cherry Valley ducks during intestinal inflammation is unknown.Three-hundred 1-day-old ducks were divided into 5 groups(6 replicates and 10 ducks per replicate)in a completely randomized design:the control and dextran sulfate sodium(DSS)groups were fed a corn-soybean meal basal diet,and experimental groups received supplements of 70,120 or 170 mg/kg Zn in form of Zn-Gly.The DSS and treatment groups were given 2 mL of 0.45 g/mL DSS daily during d 15–21,and the control group received normal saline.The experiment lasted 21 d.Results Compared with DSS group,70,120 and 170 mg/kg Zn significantly increased body weight(BW),villus height and the ratio of villus to crypt,and significantly decreased the crypt depth of jejunum at 21 d.The number of goblet cells in jejunal villi in the Zn-Gly group was significantly increased by periodic acid-Schiff staining.Compared with control,the content of intestinal permeability marker D-lactic acid(D-LA)and fluxes of fluorescein isothiocyanate(FITC-D)in plasma of DSS group significantly increased,and 170 mg/kg Zn supplementation significantly decreased the D-LA content and FITC-D fluxes.Compared with control,contents of plasma,jejunum endotoxin and jejunum pro-inflammatory factors IL-1β,IL-6 and TNF-αwere significantly increased in DSS group,and were significantly decreased by 170 mg/kg Zn supplementation.Dietary Zn significantly increased the contents of anti-inflammatory factors IL-10,IL-22 and sIgA and IgG in jejunum.Real-time PCR and Western blot results showed that 170 mg/kg Zn supplementation significantly increased mRNA expression levels of CLDN-1 and expression of OCLN protein in jejunum,and decreased gene and protein expression of CLDN-2 compared with DSS group.The 120 mg/kg Zn significantly promoted the expressions of IL-22 and IgA.Dietary Zn-Gly supplementation significantly decreased pro-inflammatory genes IL-8 and TNF-αexpression levels and TNF-αprotein expression in jejunum.Additionally,Zn significantly reduced the gene and protein expression of TLR4,MYD88 and NF-κB p65.Conclusions Zn-Gly improved duck BW and alleviated intestinal injury by regulating intestinal morphology,barrier function and gut inflammation-related signal pathways TLR4/MYD88/NF-κB p65.

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.

Gga-miRNA-181-5p family facilitates chicken myogenesis via targeting TGFBR1 to block TGF-βsignaling

MicroRNAs(miRNAs)have been demonstrated to control chicken skeletal muscle growth,however,the potential function of the miR-181-5p family in chicken myogenesis remains largely unknown.Here,our study identified the two chicken(Gallus gallus;Gga)miR-181-5p family members widely expressed in various tissues,specifically miR-181a-5p and miR-181b-5p.Besides,the breast muscles of fast-growing broilers expressed higher levels of miR-181a-5p and miR-181b-5p than those of slow-growing layers.Functionally,miR-181a-5p and miR-181b-5p both promote the expression level of myogenic factors including myogenin(MyoG),myogenic differentiation 1(MyoD1),and myosin heavy chain(MyHC),meanwhile accelerating the myotube formation of skeletal muscle satellite cells(SMSCs).Mechanistically,miR-181a-5p and miR-181b-5p directly bind to the 3′untranslated region(UTR)of the transforming growth factor beta receptor 1(TGFBR1)mRNA,further reducing the expression of TGFBR1.TGFBR1 is a key Transforming growth factor beta(TGF-β)signaling transduction receptor and had a negative function in muscle cell differentiation.Furthermore,knockdown of TGFBR1 facilitated the expression of chicken myogenic factors,boosted myotube formation,and decreased the SMAD family member 2/3(SMAD2/3)phosphorylation in chicken SMSCs.SMAD2/3 are downstream of TGF-βsignaling,and miR-181a-5p and miR-181b-5p could reduce the expression of TGFBR1 to further diminish the SMAD2/3 phosphorylation.Our findings revealed that the miR-181-5p family targets TGFBR1 to break the TGF-βsignaling transduction,which resulted in promoting chicken skeletal muscle development.

Dietary resveratrol improves antioxidant status of sows and piglets and regulates antioxidant gene expression in placenta by Keap1-Nrf2 pathway and Sirt1

Background: Resveratrol, a plant phenol, affords protection against inflammation and oxidative stress. The objective of this study was to investigate the effects of dietary resveratrol supplementation during pregnancy and lactation on the antioxidant status of sows and piglets and on antioxidant gene expression and pathway in placenta.Methods: Forty sows were allotted to 2 dietary treatments 20 d after breeding. Sows were fed a control diet and a control diet with 300 mg/kg resveratrol. Oxidative stress biomarkers and antioxidant enzymes were measured in the placenta, milk, and plasma of sows and piglets. Antioxidant gene expression and protein expression of Kelch-like ECH-associated protein 1-Nuclear factor E2-related factor 2(Keap1-Nrf2), nuclear factor kappa B-p65(NFκB-p65) and sirtuin1(Sirt1) were quantified in the placenta.Results: Dietary resveratrol increased the litter and piglets weaning weights. Antioxidant status in the milk, placenta and plasma of sows and piglets was partially improved by dietary resveratrol. In placenta, Nrf2 protein expression was increased and Keap1 protein expression was decreased by dietary resveratrol. The m RNA expression of antioxidant genes including catalase(CAT), glutathione peroxidase 1(GPX1), GPX4, superoxide dismutase 1(SOD1)and heme oxygenase 1(HO1), and phase 2 detoxification genes, including glutamate-cysteine ligase modifier(GCLM), microsomal glutathione S-transferase 1(MGST1) and UDP glucuronosyltransferase family 1 member A1(UGT1 A1), was increased by dietary resveratrol. Dietary resveratrol also increased Sirt1 and phosphorylated NFκB-p65 protein expression in the placenta. We failed to observe any influences of dietary resveratrol on pro-inflammatory cytokine levels, including those of interleukin 1β(IL-1β), IL-6, IL-8 and tumor necrosis factor α(TNF-α). However, we observed that the m RNA expression of IL-8 in placenta was reduced by maternal resveratrol. In addition, dietary resveratrol showed interactive effects with day of lactation on activities of SOD and CAT and levels of malonaldehyde(MDA) and hydrogen peroxide(H2 O2) in milk.Conclusions: Dietary resveratrol supplementation during pregnancy and lactation improves the antioxidant status of sows and piglets, which is beneficial to the reproductive performance of sows. Dietary resveratrol regulates placental antioxidant gene expression by the Keap1-Nrf2 pathway and Sirt1 in placenta.

Effect of dietary supplementation with mulberry(Morus alba L.) leaves on the growth performance,meat quality and antioxidative capacity of finishing pigs

The present study was conducted to evaluate the effect of dietary mulberry(Morus alba L.) leaves powder(MLP) supplementation on meat quality of finishing pigs.A total of 40 Duroc×Landrace×Yorkshire pigs(initial body weight of(40.5±0.63) kg) were randomly allotted into two treatments,fed either with control diet or 15% MLP diet for 85 d.The results showed that MLP diet decreased(P≤0.05) average daily gain(ADG) and increased(P<0.05) feed/gain ratio(F/G) in the finishing and whole period.MLP diet also decreased(P<0.05) carcass weight,dressing percentage,last rib and average backfat depth.However,MLP diet increased(P<0.05) intramuscular fat(IMF) content,decreased(P<0.05) shear force,cooking loss and drip loss.In addition,MLP diet increased(P<0.05) total antioxidative capacity,glutathione peroxidase and tended(P<0.10) to increase total superoxide dismutase in serum.In longissimus thoracis,myosin heavy chain(MyHC) I and IIa mRNA levels were increased(P≤0.05) for MLP diet.In conclusion,15% MLP supplementation reduced the growth performance and carcass traits,but improved meat quality of finishing pigs possibly through the change of myofiber characteristics,enhancement of antioxidative capacity and increase of IMF.

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.

Alginate oligosaccharide-induced intestinal morphology, barrier function and epithelium apoptosis modifications have beneficial effects on the growth performance of weaned pigs

Background: Alginate oligosaccharide(AOS), produced from alginate by alginate lyase-mediated depolymerisation, is a potential substitute for antibiotics and possesses growth-enhancing effects. Nevertheless, the mechanisms by which AOS regulates porcine growth remain to be elucidated. Therefore, we investigated the AOS-mediated changes in the growth performance of weaned pigs by determining the intestinal morphology, barrier function,as well as epithelium apoptosis.Methods: Twenty-four weaned pigs were distributed into two groups(n = 12) and received either a basal diet(control group) or the same diet supplemented with 100 mg/kg AOS. On d 15, D-xylose(0.1 g/kg body weight)was orally administrated to eight randomly selected pigs per treatment, and their serum and intestinal mucosa samples were collected 1 h later.Results: Our results showed that inclusion of AOS in the diet for 2 wk increased(P < 0.05) the average daily body weight gain in weaned pigs. Notably, AOS supplementation ameliorated the intestinal morphology and barrier function, as suggested by the enhanced(P < 0.05) intestinal villus height, secretory immunoglobulin A content and goblet cell counts. Compared to the control group, AOS ingestion both decreased(P < 0.05) the total apoptotic percentage and increased(P < 0.05) the proportion of S phase in the intestinal epithelial cells. Furthermore, AOS not only up-regulated(P < 0.05) the B-cell lymphoma-2(BCL2) transcriptional level but also down-regulated(P < 0.05) the B-cell lymphoma-2-associated X protein(BAX), cysteinyl aspartate-specific proteinase-3(caspase-3) and caspase-9 transcriptional levels in the small intestine.Conclusions: In summary, this study provides evidence that supplemental AOS beneficially affects the growth performance of weaned pigs, which may result from the improved intestinal morphology and barrier function,as well as the inhibited enterocyte death, through reducing apoptosis via mitochondria-dependent apoptosis.